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In JoVE (3)

Other Publications (399)

Articles by Min Zhao in JoVE

 JoVE Bioengineering

Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments

1School of Dentistry, Cardiff Institute of Tissue Engineering & Repair, Cardiff University, 2Shandong Qianfoshan Hospital, Shandong University School of Medicine, 3Dermatology and Ophthalmology Research, Institute for Regenerative Cures, University of California at Davis


JoVE 3453

This protocol demonstrates methods used to establish 2D and 3D environments in custom-designed electrotactic chambers, which can track cells in vivo/ex vivo using time-lapse recording at the single cell level, in order to investigate galvanotaxis/electrotaxis and other cellular responses to direct current (DC) electric fields (EFs).

 JoVE Biology

Measurement of Extracellular Ion Fluxes Using the Ion-selective Self-referencing Microelectrode Technique

1Department of Dermatology, Institute for Regenerative Cures, University of California, Davis, 2Departamento de Biologia, Centro de Biologia Molecular e Ambiental, Universidade do Minho, 3Department of Neurology and Center for Neuroscience, University of California, Davis Imaging of Dementia and Aging Laboratory, 4Department of Ophthalmology, Institute for Regenerative Cures, University of California, Davis


JoVE 52782

Transporters in cell membranes allow differential segregation of ions across cell membranes or cell layers and play crucial roles during tissue physiology, repair and pathology. We describe the ion-selective self-referencing microelectrode that allows the measurement of specific ion fluxes at single cells and tissues in vivo.

Other articles by Min Zhao on PubMed

Determination of Sulfite in Beer Samples Using an Amperometric Fill and Flow Channel Biosensor Employing Sulfite Oxidase

A simple method is described to determine sulfite in beer samples using a fill and flow channel biosensor. A droplet of sample is placed into the inlet of a rectangular flow cell and begins to flow through the channel by capillarity. The flow is maintained and controlled by a porous outlet plug of defined porosity. In a rectangular flow cell, the sample solution flows through three consecutive zones: over a predictor electrode, an enzyme layer and a detector electrode. Together these three zones enable the differentiation between current due to sulfite and current due to other electroactive species in the sample. The predictor electrode is located upstream, and on the opposite channel wall to the enzyme layer and detector electrode, and is poised at the same potential (+0.65 V versus Ag/AgCl) as the detector electrode. On this electrode, the current contribution from all species in the sample solution that are oxidized at that potential is determined. The enzyme layer contains sulfite oxidase, which, in the process of oxidizing sulfite, produces hydrogen peroxide, which itself is reduced by excess sulfite. The current at the downstream detector electrode is therefore different from that at the predictor electrode as a result of the enzyme reaction and the difference of the currents, corrected for the dimensions of the electrodes, is proportional to the concentration of sulfite. The method enables a straightforward correction of the interfering current at the detector electrode and a determination of the analyte concentration. The effect of interferences from ascorbic acid, ethanol, sorbic acid and tartaric acid in the detection of sulfite is efficiently removed. The concentration of sulfite in a sample of beer measured by the biosensor is equivalent to that measured using a reference method based on the AOAC-recommended Monier-Williams method.

Agreement Between Two Non-contact Specular Microscopes: Topcon SP2000P Versus Rhine-Tec

Pulmonary Macrophage Inhibition and Inhaled Nitric Oxide Attenuate Lung Ischemia-reperfusion Injury

Lung ischemia-reperfusion injury (LIRI) is postulated to occur biphasically. Donor pulmonary macrophages mediate early injury, and neutrophil-dependent injury predominates in the later phase of LIRI. We hypothesized that the biphasic response to LIRI would be attenuated by the administration of gadolinium, a known pulmonary macrophage inhibitor, and inhaled nitric oxide (NO), a pulmonary vasodilator that also interferes with neutrophil chemotaxis.

Comparison of Two Semiautomated Methods for Evaluating Endothelial Cells of Eye Bank Corneas

To compare two semiautomated methods of evaluating endothelial cells of eye bank corneas.

The Multifunctional Human P100 Protein 'hooks' Methylated Ligands

The human p100 protein is a vital transcription regulator that increases gene transcription by forming a physical bridge between promoter-specific activators and the basal transcription machinery. Here we demonstrate that the tudor and SN (TSN) domain of p100 interacts with U small nuclear ribonucleoprotein (snRNP) complexes, suggesting a role for p100 in the processing of precursor messenger RNA. We determined the crystal structure of the p100 TSN domain to delineate the molecular basis of p100's proposed functions. The interdigitated structure resembles a hook, with a hinge controlling the movement and orientation of the hook. Our studies suggest that a conserved aromatic cage hooks methyl groups of snRNPs and anchors p100 to the spliceosome. These structural insights partly explain the distinct roles of p100 in transcription and splicing.

Physical Characteristics and Aerosolization Performance of Insulin Dry Powders for Inhalation Prepared by a Spray Drying Method

The objective of this study was to investigate the influence of formulation excipients on the physical characteristics and aerosolization performance of insulin dry powders for inhalation. Insulin dry powders were prepared by a spray drying technique using excipients such as sugars (trehalose, lactose and dextran), mannitol and amino acids (L-leucine, glycine and threonine). High performance liquid chromatography and the mouse blood glucose method were used for determination of the insulin content. The powder properties were determined and compared by scanning electron microscopy, thermo-gravimetric analysis and size distribution analysis by a time-of-flight technique. The in-vitro aerosolization behaviour of the powders was assessed with an Aerolizer inhaler using a twin-stage impinger. Powder yield and moisture absorption were also determined. Results showed that there was no noticeable change in insulin content in any of the formulations by both assay methods. All powders were highly wrinkled, with median aerodynamic diameters of 2-4 microm, and consequently suitable for pulmonary administration. The tapped density was reduced dramatically when glycine was added. The powders containing mannitol, with or without L-leucine, were less sensitive to moisture. The highest respirable fraction of 67.3 +/- 1.3% was obtained with the formulation containing L-leucine, in contrast to formulations containing glycine and threonine, which had a respirable fraction of 11.2 +/- 3.9% and 23.5 +/- 2.5%, respectively. In addition, powders with good physical properties were achieved by the combination of insulin and trehalose. This study suggests that L-leucine could be used to enhance the aerosolization behaviour of the insulin dry powders for inhalation, and trehalose could potentially be used as an excipient in the formulations.

[A Case of Delayed Cervical Epidural Hematoma with C5 Nerve Root Palsy After Posterior Cervical Laminoplasty]

Delayed postoperative spinal epidural hematoma (DPSEH) is a rare and potentially devastating complication of laminoplasty, and cervical nerve root palsy occurs more frequently than DPSEH, especially with C5 nerve root palsy. The authors describe a case of DPSEH with C5 nerve root palsy that developed in a patient 3 days after he underwent laminoplasty. In this case, a 78-year-old man with a history that having taken Aspirin without the doctor's instruction for 5-6 years, he underwent cervical laminoplasty for mild myelopathy. On the 3rd postoperative day, he complained of weakness of his left upper extremity and could not raise his left arm. The symptom aggravated in the next few days. On the 9th postoperative day, there was an obvious motor deficit of both upper and lower extremities. Magnetic resonance imaging demonstrated abnormal signal characteristics consistent with a hematoma at levels C3-C4, compressing spinal cord. The clot was evacuated during emergency revision surgery, and the postoperative course after the operation was uneventful and the muscle strength was improved five days later. Therefore, the symptoms of DPSEH are not so typical that its possibility should be kept in mind. Sometimes a differential diagnosis should be made with C5 nerve root palsy which may only represent weakness of upper extremities. The authors recommend that magnetic resonance imaging is helpful for the diagnosis of DPSEH and a revision surgery should be taken as soon as possible once the hematoma causing the neurologic deterioration was confirmed.

EGF Receptor Signalling is Essential for Electric-field-directed Migration of Breast Cancer Cells

The mechanisms by which cancer cells migrate to metastasise are not fully understood. Breast cancers are accompanied by electrical depolarisation of tumour epithelial cells. The electrical changes can be detected on the skin and are used to differentiate malignant from benign breast tumours. Could the electrical signals play a role in metastasis by promoting tumour cell migration? We report that electric fields stimulate and direct migration of human breast cancer cells. Importantly, these effects were more significant in highly metastatic tumour cells than in low metastatic tumour cells. Electric-field-enhanced directional migration correlates well with the expression level of EGF receptor (EGFR/ErbB1). To confirm this, we transfected low metastatic clone MTC cells with human ErbB1, which significantly increased the electrotactic response. Inhibition of ErbB1 completely abolished the directional response of MTLn3 cells to an electric field. Transfection of MTLn3 cells and MDA-MB-435 cells with expression vectors for ErbB family members ErbB1, ErbB2 and ErbB3 also significantly enhanced EF-induced migration. These results suggest that electric signals might play a role in metastasis of breast cancers by enhancing cell migration through the ErbB-signalling pathway.

Nerve Regeneration and Functional Recovery After a Sciatic Nerve Gap is Repaired by an Acellular Nerve Allograft Made Through Chemical Extraction in Canines

The purpose of the present study is to test whether chemically extracted acellular nerve segments can be used to repair the sciatic nerve gap. Fifteen canines were divided into acellular nerve allografting group (ANG, six canines), autografting group (AG, six canines), and fresh nerve allografting group (FNG, three canines). The sciatic nerves on the right side of the animals were exposed, and 5-cm-long segments of the nerves were removed from the midthigh level and replaced by the three types of grafts. At 6 months after grafting, all animals in the ANG and AG had similar patterns of right posterior limb gait cycle and right ankle movements. Moreover, the animals in the ANG and AG had similar nerve regeneration, with dense regeneration fibers in the distal tibial nerves and obvious motor end plates in the target muscle. But in FNG, the area surrounding the graft was scarred as the result of inflammation, and there was a brown central area where there was little nerve regeneration. All of the above shows that chemical acellular nerve allografting can be used to repair a gap as long as 5 cm in the continuity of the sciatic nerve in canines and has similar effects to autografting.

Evidence for the Presence of Stem Cell-like Progenitor Cells in Human Adult Pancreas

The origin of cells replacing ageing beta-cells in adult life is unknown. This study assessed the expression of classic stem cell markers: Oct4, Sox2 and CD34 in islet-enriched fractions versus exocrine cell-enriched fractions from 25 adult human pancreases following human islet isolation. Expression of Oct4, Sox2 and CD34 mRNAs was found in all cell samples, with no significant differences between endocrine and exocrine cell fractions. Immunohistochemical staining for Oct4, Sox2, CD133, CD34, CK19, insulin and nestin on human pancreas sections showed that the majority of Oct4(+ve) cells were found in the walls of small ducts. Similar localisations were observed for Sox2(+ve) cells. The majority of Sox2(+ve) cells were found to co-express Oct4 proteins, but not vice versa. Cells positive for Oct4 and Sox2 appeared to be a unique cell population in the adult human pancreases without co-expression for CK19, CD34, CD133, insulin and nestin proteins. The numbers of Oct4(+ve) and Sox2(+ve) cells varied among donors and were approximately 1-200 and 1-30 per 100 000 pancreatic cells respectively.

[Anatomical Study on Anterior Transposition of Ulnar Nerve Accompanied with Arteries for Cubital Tunnel Syndrome]

To investigate the blood supply of the ulnar nerve in the elbow region and to design the procedure of anterior transposition of ulnar nerve accompanied with arteries for cubital tunnel syndrome.

[Application of Reversed Pedicled Submental Island Flap]

To explore the clinical applications of reversed pedicled submental island flap in the face and oropharynx.

Prejunctional Inhibitory Effects of Isoprostanes on Dopaminergic Neurotransmission in Bovine Retinae, in Vitro

We investigated the effect of isoprostanes (IsoPs) on potassium (K+)-depolarization-evoked release of [3H]dopamine from isolated bovine retinae. Isolated retinae were preloaded with [3H]dopamine and then prepared for studies of [3H]dopamine release using the superfusion method. 8-iso(15R)PGF 2alpha, 8-isoPGE2, 8-isoPGE1 and 8-isoPGF 2alpha attenuated [3H]dopamine release from isolated bovine retinae. At a concentration of 1 microM, the rank order of activity displayed by IsoP agonists was: 8-iso(15R)PGF 2alpha > 8-isoPGE2 > 8-isoPGE1 > 8-isoPGF 2alpha. Inhibition of cyclooxygenase (COX) with flurbiprofen reversed the effects caused by 8-isoPGE2 (10 nM and 10 microM), 8-iso(15R)PGF 2alpha (1 microM) and 8-isoPGE1 (1 microM). Although the EP1/EP2 antagonist, AH 6809 (10 microM) had no significant effect on K+-induced [3H]dopamine release, it blocked the inhibitory effect of both 8-isoPGE1 (10 microM) and 8-isoPGE2 (10 microM). In conclusion, IsoPs attenuate K+-induced [3H]dopamine release in isolated bovine retinae, presumably via an indirect action on COX pathway leading to the production of prostanoids, which in turn, activates EP receptors.

Expression Status of Ataxia-telangiectasia-mutated Gene Correlated with Prognosis in Advanced Gastric Cancer

Many studies have revealed the ATM alterations involved in cancer development and progression. In order to elucidate ATM deficiency in advanced GC and its clinical significance, a total of 20 exons of ATM gene, including frequently reported variations, were screened in 40 advanced primary GC and matched normal tissues using denaturing high performance liquid chromatography (DHPLC) and DNA sequencing analysis. Furthermore, ATM mRNA level was analyzed using Real-time RT-PCR and in situ hybridization, and protein expression and phosphorylation at Ser1981 were measured by immunohistochemical assessment in tissue microarray of GC. Five variants were identified in 6 of 40 cases (15%), but no hot spot of variation was detected. However, decreased expression and phosphorylation of ATM were consistently presented in tumors. In a cohort of 70 GC samples, low level of phosphorylated ATM was significantly correlated with poor differentiation, lymph node metastasis and poor 5-year survival (P<0.05). These results indicated that ATM phosphorylation status might be a prognostic marker for individual therapy in advanced GC patients.

Intracellular Ca2+ Stores Are Essential for Injury Induced Ca2+ Signaling and Re-endothelialization

Endothelialization repairs the lining of damaged vasculature and is a key process in preventing thrombosis and restenosis. It has been demonstrated that extracellular calcium ([Ca2+](o)) influx is important for subsequent endothelialization. The role of intracellular Ca2+ stores in mechanical denudation induced intracellular calcium ([Ca2+](i)) rise and endothelialization remains to be demonstrated. Using monolayer culture of a human endothelial cell line (human umbilical vein endothelial cell, HUVEC), we investigated [Ca2+](i) wave propagation and re-endothelialization following mechanical denudation. Consistent with previous reports for other types of cells, mechanical denudation induces calcium influx, which is essential for [Ca2+](i) rise and endothelialization. Moreover, we found that intracellular Ca(2+) stores are also essential for denudation induced [Ca2+](i) wave initiation and propagation, and the subsequent endothelialization. Thapsigargin which depletes intracellular Ca2+ stores completely abolished [Ca2+](i) wave generation and endothelialization. Xestospongin C (XeC), which prevents Ca2+ release from intracellular Ca2+ stores by inhibition of inositol 1,4,5-trisphosphate (IP(3)) receptor, inhibited intercellular Ca2+ wave generation and endothelialization following denudation. Purinergic signaling through a suramin sensitive mechanism and gap junction communication also contribute to in intercellular Ca(2+) wave propagation and re-endothelialization. We conclude that intracellular Ca2+ stores, in addition to extracellular Ca2+, are essential for intracellular Ca2+ signaling and subsequent endothelialization following mechanical denudation.

Delta-Opioid Receptor Stimulation Enhances the Growth of Neonatal Rat Ventricular Myocytes Via the Extracellular Signal-regulated Kinase Pathway

1. The aims of the present study were to determine whether delta-opioid receptor stimulation enhanced proliferation of and to investigate the role of the extracellular signal-regulated kinase (ERK) pathway in ventricular myocytes from neonatal rats. 2. At concentratins ranging from 10 nmol/L to 10 micromol/L, [D-Ala2,D-Leu5]enkephalin (DADLE) concentration-dependently promoted myocardial growth and DNA synthesis and altered the cytoskeleton. 3. At 1 micromol/L, DADLE also increased the expression and phosphorylation of ERK. 4. These effects of 1 micromol/L DADLE were abolished by 10 micromol/L naltrindole, a selective delta-opioid receptor antagonist, 10 nmol/L U0126, a selective ERK antagonist, 1 micromol/L staurosporine, an inhibitor of protein kinase (PK) C, and 100 micromol/L Rp-adenosine 3',5'-cyclic monophosphorothioate triethylammonium salt hydrate (Rp-cAMPS), an inhibitor of PKA. 5. In conclusion, delta-opioid receptor stimulation enhances the proliferation and development of the ventricular myocytes of neonatal rats. The ERK pathway and related signalling mechanisms, namely PKC and PKA, are involved.

Use of Poloxamers for Deswelling of Organ-cultured Corneas

Dextran T500, routinely used as a deswelling supplement in organ culture (OC), has been suspected of being toxic to corneal endothelial cells (ECs). This study was conducted to evaluate the innovative use of poloxamers compared with dextran for deswelling OC corneas.

Visualization of Fast-moving Cells in Vivo Using Digital Holographic Video Microscopy

Digital in-line holography offers some significant advantages over conventional optical holography and microscopy to image biological specimens. By combining holography with digital video microscopy, an in-line holographic video microscope is developed and is capable of recording spatial 3D holographic images of biological specimens, while preserving the time dimension. The system enables high-speed video recording of fast cell movement, such as the rapid movement of blood cells in the blood stream in vivo. This capability is demonstrated with observations of fast 3-D movement of live cells in suspension cultures in response to a gentle shake to the Petri dish. The experimental and numerical procedures are incorporated with a fast reconstruction algorithm for reconstruction of holographic video frames at various planes (z axis) from the hologram and along the time axis. The current system enables both lateral and longitudinal resolutions down to a few micrometers. Postreconstruction processing of background subtraction is utilized to eliminate noise caused by scattered light, thereby enabling visualization of, for example, blood streams of live Xenopos tadpoles. The combination of digital holography and microscopy offers unique advantages for imaging of fast moving cells and other biological particles in three dimensions in vivo with high spatial and temporal resolution.

Iron Complexes of Dendrimer-appended Carboxylates for Activating Dioxygen and Oxidizing Hydrocarbons

The active sites of metalloenzymes are often deeply buried inside a hydrophobic protein sheath, which protects them from undesirable hydrolysis and polymerization reactions, allowing them to achieve their normal functions. In order to mimic the hydrophobic environment of the active sites in bacterial monooxygenases, diiron(II) compounds of the general formula [Fe2([G-3]COO)4(4-RPy)2] were prepared, where [G-3]COO- is a third-generation dendrimer-appended terphenyl carboxylate ligand and 4-RPy is a pyridine derivative. The dendrimer environment provides excellent protection for the diiron center, reducing its reactivity toward dioxygen by about 300-fold compared with analogous complexes of terphenyl carboxylate ([G-1]COO-) ligands. An FeIIFeIII intermediate was characterized by electronic, electron paramagnetic resonance, Mössbauer, and X-ray absorption spectroscopic analyses following the oxygenation of [Fe2([G-3]COO)4(4-PPy)2], where 4-PPy is 4-pyrrolidinopyridine. The results are consistent with the formation of a superoxo species. This diiron compound, in the presence of dioxygen, can oxidize external substrates.

Small Applied Electric Fields Guide Migration of Hippocampal Neurons

Effectively directed neuron migration is critical for development and repair in the central nervous system (CNS). Endogenous electric fields (EFs) are widespread in developing and regenerating tissues and regulate a variety of cell behaviors including directed cell migration. Electrically-directed neuronal migration has not been tested previously and we show that an applied EF directs migration of hippocampal neurons toward the cathode at a field strength of 120 mV/mm, close to the physiological range. Reversal of the field polarity reversed the direction of neuron migration. Neuron migration from an explant also was directed by an applied EF. Mechanistically, EF-guided migration was transduced by activation of the second messenger molecules ROCK (Rho-associated protein kinase) and PI3 kinase (phosphoinositide-3 kinase) since their pharmacological inhibition decreased the directedness and speed of neuron migration. This work demonstrates that rat hippocampal neurons respond to applied EFs with directional migration and raises the possibility that EFs may be used as a cue to direct neuronal migration in novel strategies to repair the CNS.

[Peginterferon Alpha-2a and Ribavirin for Treating Chronic Hepatitis C Patients with Persistent Normal Aminotransferase Levels]

Basolateral Junctions Utilize Warts Signaling to Control Epithelial-mesenchymal Transition and Proliferation Crucial for Migration and Invasion of Drosophila Ovarian Epithelial Cells

Fasciclin2 (Fas2) and Discslarge (Dlg) localize to the basolateral junction (BLJ) of Drosophila follicle epithelial cells and inhibit their proliferation and invasion. To identify a BLJ signaling pathway we completed a genomewide screen for mutants that enhance dlg tumorigenesis. We identified two genes that encode known BLJ scaffolding proteins, lethal giant larvae (lgl) and scribble (scrib), and several not previously associated with BLJ function, including warts (wts) and roughened eye (roe), which encode a serine-threonine kinase and a transcription factor, respectively. Like scrib, wts and roe also enhance Fas2 and lgl tumorigenesis. Further, scrib, wts, and roe block border cell migration, and cause noninvasive tumors that resemble dlg partial loss of function, suggesting that the BLJ utilizes Wts signaling to repress EMT and proliferation, but not motility. Apicolateral junction proteins Fat (Ft), Expanded (Ex), and Merlin (Mer) either are not involved in these processes, or have highly spatio-temporally restricted roles, diminishing their significance as upstream inputs to Wts in follicle cells. This is further indicated in that Wts targets, CyclinE and DIAP1, are elevated in Fas2, dlg, lgl, wts, and roe cells, but not Fat, ex, or mer cells. Thus, the BLJ appears to regulate epithelial polarity and dynamics not only as a localized scaffold, but also by communicating signals to the nucleus. Wts may be regulated by distinct junction inputs depending on developmental context.

[Planting and Biological Character of Rabbit Corneal Epithelial Cells on Amniotic Membrane]

This study was aimed at investigating the cultivation and biological character of corneal epithelial cells (ECs) planted on intact and denued amniotic membrane (AM) as a substrate and trying to find out satisfactory methods for the reconstruction of corneal epithelium using tissue engineering. Rabbit corneal epithelial cells were planted on denuded AM and intact AM respectively. The cultivated corneal epithelial sheet was examined by use of inverted microscope, HE staining pathologyical section and transmission electron microscopy (TEM), and was also detected immunohistochemically. The results revealed that rabbit corneal ECs grew slowly and were difficult to stick and converge on intact AM, whereas they were easy to grow and proliferate on denuded AM. The cultivated corneal ECs showed four to five layers of stratification composed of the basement membrane of AM and multiple layers of corneal ECs showed the presence of CK3. TEM unveiled that the multiple layers of corneal ECs had numerous desmosomal junctions attaching to the basement membrane with hemidesmosomes. Therefore, the above cultivated corneal epithelial sheet can be used as engineering tissue for ocular surface reconstruction.

[Observation on Therapeutic Effect of Acupuncture Combined with He-Ne Laser Radiation on Facial Paralysis]

To compare therapeutic effects of acupuncture combined with He-Ne laser radiation and western medicine on facial paralysis.

Apoptosis of the Thick Ascending Limb Results in Acute Kidney Injury

Ischemia- or toxin-induced acute kidney injury is generally thought to affect the cells of the proximal tubule, but it has been difficult to define the involvement of other tubular segments because of the widespread damage caused by ischemia/reperfusion or toxin-induced injury in experimental models. For evaluation of whether thick ascending limb (TAL)-specific epithelial injury results in acute kidney injury, a novel transgenic mouse model that expresses the herpes simplex virus 1 thymidine kinase gene under the direction of the TAL-specific Tamm-Horsfall protein promoter was generated. After administration of gancyclovir, these mice demonstrated apoptosis only in TAL cells, with little evidence of neutrophil infiltration. Compared with control mice, blood urea nitrogen and creatinine levels were at least five-fold higher in the transgenic mice, which also developed oliguria and impaired urinary concentrating ability. These findings suggest that acute injury targeted only to the TAL is sufficient to cause severe acute kidney injury in mice with features similar to those observed in humans.

Psychological Stress Induces Hypoferremia Through the IL-6-hepcidin Axis in Rats

Anemia is a widespread public health problem. The psychological stress decreases serum iron level and inhibits erythropoiesis. However, the molecular mechanisms involved, leading to iron mal-regulation are not well known. We used a communication box paradigm to induce psychological stress and found that serum iron level decreased after 3d while liver iron storage increased after 7d. Moreover, psychological stress up-regulated expressions of interleukin-6 (IL-6) and hepcidin, while down-regulating ferroportin expression after 3d. These changes were blocked by the injection of IL-6 monoclonal antibody. In conclusion, the IL-6-hepcidin axis is up-regulated by psychological stress in rats, resulting in hypoferremia and increase of hepatic iron storage.

Study on the Relationship Between Cadmium Chloride-induced Adrenocortical Cell of Guinea Pig Apoptosis and Stress-activated Protein Kinase Activity

Heavy metals are important environmental pollutants that affect many cellular functions. The signaling pathways that lead to apoptosis of adrenocortical cells following exposure to cadmium chloride (CdCl2) have not yet been fully clarified. We developed a primary culture of guinea pig adrenocortical cells and treated it with various concentrations of CdCl2 for different time periods. The apoptosis induced by CdCl2 was detected by fluorescein isothiocyanate-labeled annexin V and propidium iodide staining using a flow cytometer. Stress-activated protein kinase (SAPK) activities were measured by immunoprecipitation and chemiluminescence assay. After 2h of treatment, the apoptotic cell rate increased in a dose-dependent manner. The difference between the high-dose group and the control group was significant (P<0.01). The apoptosis was also found to increase in a time-dependent manner in cells treated with 50micromol/L CdCl2. Among the various treatment period groups, the difference between more than 1h-treated groups and the control group was significant (P<0.01). The SAPK activity increased with an increase in CdCl2 dose after 5min of exposure. However, after 15min of exposure to CdCl2, the SAPK activity decreased with an increase in exposure time. Our findings suggest that the SAPK signaling pathway might play an important role in CdCl2-induced apoptosis of adrenocortical cells.

Amelioration of Streptozotocin-induced Diabetes in Mice with Cells Derived from Human Marrow Stromal Cells

Pluri-potent bone marrow stromal cells (MSCs) provide an attractive opportunity to generate unlimited glucose-responsive insulin-producing cells for the treatment of diabetes. We explored the potential for human MSCs (hMSCs) to be differentiated into glucose-responsive cells through a non-viral genetic reprogramming approach.

[A Comparative Study on Immunocompatibility and Histological Turnover After Fresh or Preserved Human Amniotic Membrane Xenotransplantation]

To investigate the immunoreaction, histological reaction and turnover by comparing the xenotransplantation of fresh human amniotic membrane (HAM) with that of preserved HAM, and to analyze the clinical application value of different kinds of HAM preparations.

Increased Dietary NaCl Induces Renal Medullary PGE2 Production and Natriuresis Via the EP2 Receptor

A high-NaCl diet induces renal medullary cyclooxygenase (COX)2 expression, and selective intramedullary infusion of a COX2 inhibitor increases blood pressure in rats on a high-salt diet. The present study characterized the specific prostanoid contributing to the antihypertensive effect of COX2. C57BL/6J mice placed on a high-NaCl diet exhibited increased medullary COX2 and microsomal prostaglandin E synthase1 (mPGES1) expression as determined by immunoblot and real-time PCR. Cytosolic prostaglandin E synthase and prostacyclin synthase were not induced by the high-salt diet. Immunofluorescence showed mPGES1 in collecting ducts and interstitial cells. High salt increased renal medullary PGE(2) as determined by gas chromatography/negative ion chemical ionization mass spectrometry. The effect of direct intramedullary PGE(2) infusion was examined in anesthetized uninephrectomized mice. Intramedullary PGE(2) infusion (10 ng/h) increased urine volume (from 3.3 +/- 0.6 to 9.5 +/- 1.6 mul/min) and urine sodium excretion (0.11 +/- 0.02 to 0.32 +/- 0.05 mueq/min). To determine which E-prostanoid (EP) receptor(s) mediated PGE(2)- dependent natriuresis, EP-selective prostanoids were infused. The EP(2) agonist butaprost produced natriuresis (from 0.06 +/- 0.02 to 0.32 +/- 0.05 mueq/min). The natriuretic effect of intramedullary PGE(2) or butaprost was abolished in EP2-deficient mice, which exhibit NaCl-dependent hypertension. In conclusion, a high-salt diet increases renal medullary COX2 and mPGES1 expression, and increases renal medullary PGE(2) synthesis. Renal medullary PGE(2) promotes renal sodium excretion via the EP2 receptor, thereby maintaining normotension in the setting of high salt intake.

Enhancement of Spinal N-methyl-D-aspartate Receptor Function by Remifentanil Action at Delta-opioid Receptors As a Mechanism for Acute Opioid-induced Hyperalgesia or Tolerance

Intraoperative remifentanil infusions have been associated with postoperative opioid-induced hyperalgesia and tolerance. Using a previously identified subpopulation of spinal neurons that displays an augmentation in N-methyl-D-aspartate (NMDA) receptor current after chronic morphine, investigations were undertaken to determine whether remifentanil induces acute increases in NMDA responses that are concentration dependent and receptor subtype dependent.

Intragenic Suppressors of Temperature-sensitive Rne Mutations Lead to the Dissociation of RNase E Activity on MRNA and TRNA Substrates in Escherichia Coli

RNase E of Escherichia coli is an essential endoribonuclease that is involved in many aspects of RNA metabolism. Point mutations in the S1 RNA-binding domain of RNase E (rne-1 and rne-3071) lead to temperature-sensitive growth along with defects in 5S rRNA processing, mRNA decay and tRNA maturation. However, it is not clear whether RNase E acts similarly on all kinds of RNA substrates. Here we report the isolation and characterization of three independent intragenic second-site suppressors of the rne-1 and rne-3071 alleles that demonstrate for the first time the dissociation of the in vivo activity of RNase E on mRNA versus tRNA and rRNA substrates. Specifically, tRNA maturation and 9S rRNA processing were restored to wild-type levels in each of the three suppressor mutants (rne-1/172, rne-1/186 and rne-1/187), while mRNA decay and autoregulation of RNase E protein levels remained as defective as in the rne-1 single mutant. Each single amino acid substitution (Gly-->Ala at amino acid 172; Phe --> Cys at amino acid 186 and Arg --> Leu at amino acid 187) mapped within the 5' sensor region of the RNase E protein. Molecular models of RNase E suggest how suppression may occur.

Foot-shock Stress-induced Regional Iron Accumulation and Altered Iron Homeostatic Mechanisms in Rat Brain

Like in other organs, iron in the brain plays an important role in various biological processes. Previous studies have shown that systemic iron homeostasis in mammalians was changed under specific stress conditions. The present study aimed to investigate effects of stress on brain iron homeostasis in rats using a foot-shock stress model. Young adult male Sprague-Dawley rats were randomly assigned to foot-shock stress group subjected to 30 min of cutaneous foot-shock (0.80 mA, 1 pulse/s, 300 ms duration) daily for 1 week or control group left undisturbed. Then, the rats were sacrificed and iron concentration in serum, liver, and some brain regions were measured using atomic absorption spectrophotometry. Expression of ferritin, Transferrin receptor (TfR), divalent metal transporter 1 (DMT1, with or without iron-responsive element), lactoferrin (Lf), and iron regulatory protein 1 (IRP1) in rat hippocampus were determined using western blot analysis. The results showed that stress induced decreased serum iron concentration, increased liver iron content, and elevated iron contents in specific brain regions including hippocampus, striatum, and frontal cortex. In the hippocampus, stress caused decreased expression of ferritin, increased expression of TfR and IRP1, and no change in expression of DMT1 or Lf. Results of this study demonstrated that foot-shock stress induced region specific iron accumulation and altered iron homeostatic mechanisms in the brain in addition to a changed systemic iron homeostasis characterized by decreased serum iron concentration and increased liver iron content. And, elevated IRP1 expression might be associated with the increased TfR and decreased ferritin expression, leading to subsequent iron accumulation and possible increased vulnerability to oxidative damage in hippocampus.

[Relationship Between Apoptosis and Activity of Protein Kinase B in Adrenocortical Cells Induced by Cadmium Chloride]

To study apoptosis induced by cadmium chloride (CdCl2) and the alteration in activity of protein kinase B (PKB/Akt) in adrenocortical cells.

CTLs Are Targeted to Kill Beta Cells in Patients with Type 1 Diabetes Through Recognition of a Glucose-regulated Preproinsulin Epitope

The final pathway of beta cell destruction leading to insulin deficiency, hyperglycemia, and clinical type 1 diabetes is unknown. Here we show that circulating CTLs can kill beta cells via recognition of a glucose-regulated epitope. First, we identified 2 naturally processed epitopes from the human preproinsulin signal peptide by elution from HLA-A2 (specifically, the protein encoded by the A*0201 allele) molecules. Processing of these was unconventional, requiring neither the proteasome nor transporter associated with processing (TAP). However, both epitopes were major targets for circulating effector CD8+ T cells from HLA-A2+ patients with type 1 diabetes. Moreover, cloned preproinsulin signal peptide-specific CD8+ T cells killed human beta cells in vitro. Critically, at high glucose concentration, beta cell presentation of preproinsulin signal epitope increased, as did CTL killing. This study provides direct evidence that autoreactive CTLs are present in the circulation of patients with type 1 diabetes and that they can kill human beta cells. These results also identify a mechanism of self-antigen presentation that is under pathophysiological regulation and could expose insulin-producing beta cells to increasing cytotoxicity at the later stages of the development of clinical diabetes. Our findings suggest that autoreactive CTLs are important targets for immune-based interventions in type 1 diabetes and argue for early, aggressive insulin therapy to preserve remaining beta cells.

[Decompression and Anterior Transposition of Ulnar Nerve with Inferior Ulnar Collateral Artery for Cubital Tunnel Syndrome]

To report the operation method and the clinical effect of decompression and anterior transposition of the ulnar nerve with inferior ulnar collateral artery for cubital tunnel syndrome.

[Study on the Expressed Oil of Princes-feather Fruit Before and After Processed by GC-MS]

To compare the chemical compositions in the expressed oil before and after processed of princes-feather fruit.

Novel Synthetic Luteolin Analogue-caused Sensitization of Tumor Necrosis Factor-alpha-induced Apoptosis in Human Tumor Cells

Studies on the sensitization, by novel alkynyl luteolin analogues, of TNF-alpha-induced apoptosis in HeLa and HepG2 cells revealed that LA-12 showed better sensitizing effects on TNF-alpha-induced cell death than luteolin, suggesting great potential for alkynyl luteolin analogues in cancer therapy.

Association Between Polymorphisms of FCRL3, a Non-HLA Gene, and Behçet's Disease in a Chinese Population with Ophthalmic Manifestations

Studies have shown a strong association of human leukocyte antigens-B51 (HLA-B51) with Behçet's disease (BD). However, little is known about the association of non-HLA genes with BD. The polymorphisms of the Fc receptor-like 3 gene (FCRL3), -169C/T, -110A/G, +358C/G, and +1381A/G, have been reported to be associated with several autoimmune diseases. This study was designed to determine whether the polymorphisms of FCRL3 were associated with susceptibility to BD in a Chinese population mainly with ocular involvement.

Regulation of Retinal Morphology and Posterior Segment Amino Acids by 8-isoprostaglandin E2 in Bovine Eyes Ex Vivo

There is evidence that isoprostanes (IsoPs) can regulate exogenously applied excitatory amino acid neurotransmitters in bovine retina in vitro. However, the regulation of retinal morphology and endogenous neurotransmitter levels by IsoPs is unknown. We examined the effects of intravitreally injected 8-iso-PGE(2) on retinal tissue integrity and viability and amino acid neurotransmitters in bovine eye organ culture ex vivo. Exposure of bovine eyeballs to simulated experimental conditions revealed no retinal apoptosis and necrosis in TUNEL and DAPI staining and hematoxylin and eosin staining assays, respectively, and no changes in basal levels of amino acids in retina and vitreous humor. Furthermore, intravitreal injection of 8-iso-PGE(2) into bovine eyeballs had no effect on retinal apoptosis and integrity. Interestingly, 8-iso-PGE(2) caused a concentration-dependent attenuation of retinal glutamate and its metabolite glutamine and glycine levels, while GABA was unaffected. 8-Iso-PGE(2) (1 and 100 microM) significantly (P < 0.001) attenuated glutamate levels by 33.9% and 48.0%, respectively. 8-Iso-PGE(2) (100 microM) inhibited (P < 0.01) retinal glutamine and glycine levels by 37.7% and 35.5%, respectively. The IsoP exhibited no effect on vitreous humor glutamine and glycine levels, while glutamate and GABA were not detected. Thus, 8-iso-PGE(2) can regulate retinal amino acids without inducing cell death in bovine retina ex vivo.

Regulation of Neurotransmitter Release from Ocular Tissues by Isoprostanes

Isoprostanes are prostaglandin-like compounds formed in vivo primarily by free radical-catalyzed peroxidation of arachidonic acid independent of the cyclooxygenase enzyme. In addition to being utilized as reliable indicators of oxidative stress, 8-isoprostanes exert pharmacological actions on smooth muscles from several tissues and organs, and they play a role in the release of neurotransmitters from the central and peripheral nervous systems. In the anterior uvea of the eye, 8-isoprostanes produce both excitatory and inhibitory effects on sympathetic neurotransmission in isolated mammalian iris ciliary bodies. Thromboxane (TP) receptors mediate the stimulatory action of isoprostanes on norepinephrine (NE) release from sympathetic nerves. In bovine retina, the 8-isoprostanes exhibit a biphasic regulatory effect on potassium-induced [3H]-D-aspartate release, with low concentrations being inhibitory and high concentrations causing an excitatory effect. Excitatory effects of 8-isoprostanes are mediated by TP receptors, while inhibitory responses are mediated by prostaglandin E (EP) receptors. The 8-isoprostanes produce pharmacological actions on sympathetic neurotransmission in mammalian anterior uvea, a response that is species-dependent. In the posterior segment of the eye, 8-isoprostanes elicit a complex response on the retina involving the activation of both prostanoid TP and EP receptors. An effect of isoprostanes on neurotransmitter pools provides new pharmacological target sites for the therapy of some ocular diseases.

[Development and Clinical Application of Frozen Tissue Microarray in Lung Cancer Diagnosis.]

Traditional techniques in clinical diagnostic pathology have some limitations. Here we tried to develope frozen tissue microarray as a fast, simple and economical technique.

2-Butyl-1,3-diphenyl-2,3-dihydro-1H-naphtho[1,2-e][1,3]oxazine

In the title compound, C(28)H(27)NO, the oxazine ring adopts a half-chair conformation. The dihedral angles between the phenyl rings and the naphthyl ring system are 15.34 (1) and 76.51 (1)°.

1-{Phen-yl[1-(p-tol-yl)ethyl-amino]meth-yl}-2-naphthol

The title compound, C(26)H(25)NO, was obtained via a one-pot synthesis from the reaction of 2-naphthol, 1-(p-tol-yl)ethyl-amine, p-toluene-sulfonic acid and benzaldehyde. There are three mol-ecules per asymmetric unit, all having similar conformations. There are intra-molecular O-H⋯N and C-H⋯O hydrogen bonds, with only van der Waals forces found between mol-ecules.

Carbon Nanotube-enhanced DNA Biosensor for DNA Hybridization Detection Using Manganese(II)-Schiff Base Complex As Hybridization Indicator

A Mn(II) complex, MnL (L=sodium (E)-3-((1-carboxyethylimino)methyl)-4-hydroxybenzenesulfonate), was synthesized and characterized using elemental analysis and IR spectroscopy. Cyclic voltammetry (CV) and fluorescence spectroscopy were used to investigate the interaction between MnL and salmon sperm DNA. It was revealed that MnL presented high electrochemical activity on glassy carbon electrode (GCE), and it could be intercalated into the double helices of double-stranded DNA (dsDNA). Using MnL as the hybridization indicator, a novel and sensitive electrochemical DNA biosensor based on multiwall carbon nanotubes functionalized with carboxyl groups (MWCNTs-COOH, on which DNA probes were covalently immobilized) was prepared. The target single-stranded DNA (ssDNA) could be quantified ranging from 6.7 x 10(-10)M to 8.4 x 10(-9)M with good linearity (r=0.9922). A detection limit of 1.4 x 10(-10)M (3 sigma, n=9) was achieved.

[Effect of Change in High Mobility Group Protein Box 1 Expression on Activity of Immunocytes in Spleen of Mice with Multiple Organ Dysfunction Syndrome]

To explore the regularity of high mobility group protein box 1 (HMGB1) expression and major histocompatibility complex II I-A(b) in spleen of mice with multiple organ dysfunction syndrome (MODS) , and its effect on the activity of immunocytes and relationship with pathogenesis of MODS.

Electrical Fields in Wound Healing-An Overriding Signal That Directs Cell Migration

Injury that disrupts an epithelial layer instantaneously generates endogenous electric fields (EFs), which were detected at human skin wounds over 150 years ago. Recent researches combining molecular, genetic and imaging techniques have provided significant insights into cellular and molecular responses to this "unconventional" signal. One unexpected finding is that the EFs play an overriding guidance role in directing cell migration in epithelial wound healing. In experimental models where other directional cues (e.g., contact inhibition release, population pressure etc.) are present, electric fields of physiological strength override them and direct cell migration. The electrotaxis or galvanotaxis is mediated by polarized activation of multiple signaling pathways that include PI3 kinases/Pten, membrane growth factor receptors and integrins. Genetic manipulation of PI3 kinase/Pten (Phosphoinositide 3-kinases/phosphatase and tensin homolog) and integrin beta4 demonstrated the importance of those molecules. The electric fields are therefore a fundamental signal that directs cell migration in wound healing. One of the most challenging question is: How do cells sense the very weak electric signals? Clinically, it is highly desirable to develop practical and reliable technologies for wound healing management exploiting the electric signaling.

Conformation-dependent Single-chain Variable Fragment Antibodies Specifically Recognize Beta-amyloid Oligomers

Increasing evidence indicates that beta-amyloid (Abeta) oligomers rather than monomers or fibrils are the major toxic agents that specifically inhibit synaptic plasticity and long-term potentiation (LTP) in Alzheimer's disease (AD). Neutralization of Abeta oligomeric toxicity was found to reverse memory deficits. Here, we report four single-chain variable fragment (scFv) antibodies isolated from the naive human scFv library by phage display that specifically recognized Abeta oligomers but not monomers and fibrils. These conformation-dependent scFv antibodies inhibit both Abeta fibrillation and cytotoxicity and bind to the same type of eptitope displayed on the Abeta oligomers. Such scFv antibodies specifically targeting toxic Abeta oligomers may have potential therapeutic and diagnostic applications for AD.

Electrical Signals Polarize Neuronal Organelles, Direct Neuron Migration, and Orient Cell Division

During early brain development, the axis of division of neuronal precursor cells is regulated tightly and can determine whether neurons remain in the germinal layers or migrate away. Directed neuronal migration depends on the establishment of cell polarity, and cells are polarized dynamically in response to extracellular signals. Endogenous electric fields (EFs) orient cell division and direct migration of a variety of cell types. Here, we show that cell division of cultured hippocampal cells (neuron-like cells and glial-like cells) is oriented strikingly by an applied EF, which also directs neuronal migration. Directed migration involves polarization of the leading neurite, of the microtubule-associated protein MAP-2 and of the Golgi apparatus and the centrosome, all of which reposition asymmetrically to face the cathode. Pharmacological inhibition of Rho-associated coiled-coil forming protein kinases (ROCK) and phosphoinositide 3-kinase decreased, leading neurite orientation and Golgi polarization in the neurons in response to an EF and in parallel decreased the directedness of EF-guided neuronal migration. This work demonstrates that the axis of hippocampal cell division, the establishment of neuronal polarity, the polarization of intracellular structures, and the direction of neuronal migration are all regulated by an extracellular electrical cue.

[Experimental Investigation of PEGFP-bFGF Gene Transfer to Human Limbal Stem Cells]

Primary HLSCs were successfully cultured and assayed by AE5 in vitro. Constructed eukaryotic expressive vector of pEGFP-bFGF was transferred into the human limbal stem cells by the liposome-mediated technique, and 48 hours later, specific green fluorescence was observed by fluorescence microscope. The gene transfeetion efficiency was 20%-30%. Then the model of cells injury was created by use of NaOH. The cells were divided into four groups: Normal, bFGF, NaOH and bFGF+NaOH. The cellular viability in each group was measured by MTT colorimetry, and the cellular apoptosis rate and necrosis rate were observed by laser scanning confocal microscopy. The cellular viability in bFGF+NaOH group was higher than that in NaOH group (P < 0.05) ,while the cellular apoptosis rate plus necrosis rate displayed significant difference between the two groups (P < 0.05). The pEGFP-bbGF gene was noted to be successfully transferred into HLSCs and the cells were found growing well. These indicated that bFGF gene has a protective effect on the HLSCs injured by NaOH. We have also probed the feasibility of trying the treatment for ocular surface disease through gene engineering recombined tissue engineering.

Progressive CD127 Down-regulation Correlates with Increased Apoptosis of CD8 T Cells During Chronic HIV-1 Infection

Chronic HIV-1 infection can induce a significant decrease in CD127 expression on CD8 T cells, but the underlying mechanisms and immunological consequences are unclear. In this study, we investigated CD127 expression on CD8 T cells from a total of 51 HIV-1-infected subjects and 16 healthy individuals and analyzed the association between CD127 expression and CD8 T-cell apoptosis in these HIV-1-infected subjects. We found that CD127 expression on total CD8 T cells was significantly down-regulated, which was correlated with the increased CD8 T-cell apoptosis and disease progression of chronic HIV-1 infection. The in vitro addition of IL-7 efficiently rescued the spontaneous apoptosis of CD8 T cells from HIV-1-infected individuals. IL-7 stimulation also transiently down-regulated CD127 expression, whereas some of the CD127(-) CD8 T cells regained CD127 expression soon after IL-7 was retracted from the incubation medium. Thus, IL-7 stimulation reduced apoptosis of both CD127(+) and CD127(-)CD8 T cells to some degree. These data indicate that CD127 loss might impair IL-7 signaling and increase CD8 T-cell apoptosis during HIV-1 infection. This study, therefore, will extend the notion that IL-7 could be a good candidate for immunotherapy in HIV-1-infected patients.

N546 in Beta18-beta19 Loop is Important for Binding and Toxicity of the Bacillus Thuringiensis Cry1Ac Toxin

Our previous mutagenic analysis showed that the unique residue N546 in the apex of beta18-beta19 loop of Bacillus thuringiensis Cry1Ac toxin is important for its toxicity. In this study, trypsin digestion susceptibility, binding to BBMV and oligomer formation activity was therefore analyzed to determine the mechanism of toxicity change of these mutant toxins. The results showed that residue N546 was not involved in toxin oligomerisation and maintaining the stability of toxin, the enhanced toxicity of mutant N546A was just because of increased binding to BBMV, and reduction in toxicity of other mutants were caused by reduction in initial or irreversible binding to BBMV. This is the first report that revealed N546 in Cry1Ac domain III played an essential role in its insecticidal activity and binding to insect BBMV.

[HEV Capsid Protein Interacts with CYP 2A6 and Decreases Its Coumarin 7-hydroxylation Activity]

E2 is a recombinant hepatitis E virus capsid protein including its main antigenic determinants but lacking of the particle assembling domain. P239 was the C-terminal extending protein of E2 and could self-assemble to form virus like particles, which might serve as mimicry of virions both structurally and antigenically. We previously used yeast two-hybrid system to screen proteins interacting with E2 based on a human hepatocyte cDNA library. One candidate was identified as the segment (aa388-437) of cytochrome P450 2A6 protein, which is predominantly expressed in liver and important for metabolization. Some studies have demonstrated that hepatitis virus infection may altered cell metabolic clearance of coumrarin which were rapidly matebolised by CYP2A6. In this research, we demonstrated that the protein interaction between HEV capsid proteins and CYP2A6 by pull-down and co-immunoprecipitation. It was also found that their interaction could decrease the CYP2A6 catalytic activity when p239 was incubated within the CYP2A6-transfected Huh7 cells. These results suggested that CYP2A6 might be related to the pathological process when HEV invaded host cells.

Polymorphisms of FCRL3 in a Chinese Population with Vogt-Koyanagi-Harada (VKH) Syndrome

The polymorphisms of the Fc receptor-like 3 gene (FCRL3), a novel immunoregulatory gene, have been shown to be associated with certain autoimmune diseases. This study was designed to examine whether the polymorphisms of FCRL3 are associated with susceptibility to Vogt-Koyanagi-Harada (VKH) syndrome in a Chinese population.

RLEdb: a Database of Rate-limiting Enzymes and Their Regulation in Human, Rat, Mouse, Yeast and E. Coli

[Study on the Expression and Significance of Galectin-3 and CDC25B MRNA in Human Gastric Carcinoma]

To study the expression of Galectin-3 and CDC25B mRNA in gastric carcinoma and their correlation with clinical-pathological features and the survival time.

Role of Prostanoid Production and Receptors in the Regulation of Retinal Endogenous Amino Acid Neurotransmitters by 8-isoprostaglandin E2, Ex Vivo

The role of enzymes and receptors of the prostanoid pathway in the inhibitory effect of 8-isoprostaglandin E2 (8-isoPGE2) on endogenous amino acid neurotransmitter levels was examined, ex vivo. Freshly isolated bovine eyeballs were injected intravitreally with IsoPs, incubated in Krebs buffer for 30 min and retina prepared for HPLC-ECD detection of amino acids. 8-isoPGE2 attenuated retinal glutamate and its metabolite, glutamine and glycine in a concentration-dependent manner. The nonselective cyclooxygenase (COX)-inhibitor, flurbiprofen, COX-2 selective inhibitor, NS-398 and thromboxane (Tx) synthase inhibitor, furegrelate had no effect on both basal amino acid levels and the inhibitory effects of 8-isoPGE2 (1-100 μM) on the retinal amino acids. Whereas the TP-receptor antagonist SQ-29548(10 μM) exhibited no effect, SC-19220(EP1; 30 μM), AH-6809(EP(1-3); 30 μM) and AH-23848(EP4; 30 μM) reversed the inhibitory effects of 8-isoPGE2 (0.01-100 μM) on glutamate, glutamine and glycine levels. We conclude that prostanoid EP-receptors regulate the inhibitory effect of 8-isoPGE2 on basal levels of endogenous amino acids in bovine retina, ex vivo.

[Trilobate Technique, a New Principal to Repair Cleft Lip]

To develop a new method for reparation of cleft lip, and to evoke more colleagues for advance practices and study, in order to determine her indication and contraindication as soon as possible.

Purity Determination and Uncertainty Evaluation of Theophylline by Mass Balance Method, High Performance Liquid Chromatography and Differential Scanning Calorimetry

Purity determination of pure organic substance is essential for the establishment of traceability to SI units. A mass balance method was employed to determine the purity of theophylline certified reference materials (CRM), compared with high performance liquid chromatography (HPLC) and differential scanning calorimetry (DSC). In the approach of the mass balance, the impurities were identified by ion trap time-of-flight mass spectrometer (IT-TOF-MS) and quantified by HPLC. The purity of theophylline CRM determined by mass balance method was 99.82% with an extended uncertainty of 0.1% (k=2). The uncertainty evaluation of purity demonstrated that the accuracy of the mass balance method is better than that of HPLC and DSC. It indicated that the mass balance is suitable for the CRM and pharmaceutical standards.

[The Role of FDG-PET in Staging of Lymphoma and Evaluation of Therapeutic Efficiency]

To evaluate the application of 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) to the staging and detecting residual masses of lymphoma.

Electric Currents in Xenopus Tadpole Tail Regeneration

Xenopus laevis tadpoles can regenerate tail, including spinal cord, after partial amputation, but lose this ability during a specific period around stage 45. They regain this ability after stage 45. What happens during this "refractory period" might hold the key to spinal cord regeneration. We hypothesize that electric currents at amputated stumps play significant roles in tail regeneration. We measured electric current at tail stumps following amputation at different developmental stages. Amputation induced large outward currents leaving the stump. In regenerating stumps of stage 40 tadpoles, a remarkable reversal of the current direction occurred around 12-24 h post-amputation, while non-regenerating stumps of stage 45 tadpole maintained outward currents. This reversal of electric current at tail stumps correlates with whether tails regenerate or not (regenerating stage 40-inward current; non-regenerating stage 45-outward current). Reduction of tail stump current using sodium-free solution decreased the rate of regeneration and percentage regeneration. Fin punch wounds healed normally at stages 45 and 48, and in sodium-free solution, suggesting that the absence of tail re-growth at stage 45 is regeneration-specific rather than a general inhibition of wound healing. These data suggest that electric signals might be one of the key players regulating regeneration.

Resveratrol Inhibits Beta-amyloid Oligomeric Cytotoxicity but Does Not Prevent Oligomer Formation

Beta-amyloid (Abeta) aggregation has been strongly associated with the neurodegenerative pathology and a cascade of harmful event rated to Alzheimer's disease (AD). Inhibition of Abeta assembly, destabilization of preformed Abeta aggregates and attenuation of the cytotoxicity of Abeta oligomers and fibrils could be valuable therapeutics of patients with AD. Recent studies suggested that moderate consumption of red wine and intake of dietary polyphenols, such as resveratrol, may benefit AD phenotypes in animal models and reduce the relative risk for AD clinical dementia. To understand the mechanism of this neuroprotection, we studied the effects of resveratrol, an active ingredient of polyphenols in wine and many plants, on the polymerization of Abeta42 monomer, the destabilization of Abeta42 fibril and the cell toxicity of Abeta42 in vitro using fluorescence spectroscopic analysis with thioflavin T (ThT), transmission electron microscope (TEM), circular dichroism (CD) and MTT assay. The results showed that resveratrol could dose-dependently inhibit Abeta42 fibril formation and cytotoxicity but could not prevent Abeta42 oligomerization. The studies by Western-blot, dot-blot and ELISA confirmed that the addition of resveratrol resulted in numerous Abeta42 oligomer formation. In conjunction with the concept that Abeta oligomers are linked to Abeta toxicity, we speculate that aside from potential antioxidant activities, resveratrol may directly bind to Abeta42, interfere in Abeta42 aggregation, change the Abeta42 oligomer conformation and attenuate Abeta42 oligomeric cytotoxicity.

Electrotaxis and Wound Healing: Experimental Methods to Study Electric Fields As a Directional Signal for Cell Migration

Electric fields were measured at human skin wounds over one and half centuries ago. Modern techniques have verified and greatly extended our understanding of the existence of endogenous wound electric fields. In virtually all wounds studied, disruption of an epithelial layer instantaneously generates endogenous electric fields. As electric fields have the intrinsic property of being vectorial, it has long been proposed that these fields may serve as a directional signal guiding cell migration in wound healing. We have established several experimental systems to study the guidance effects and mechanisms of electric fields on cell migration. Most types of cells migrate directionally in a small electric field, a phenomenon called galvanotaxis/electrotaxis. Remarkably, electric fields of strength equal to those detected at in vivo wounds direct cell migration and override some other well-accepted coexistent guidance cues such as contact inhibition. The naturally occurring endogenous electric fields therefore may be an important signaling mechanism that regulates directional cell movement in vivo. Applied electric fields may have a potential clinical role in guiding cell migration in wound healing. The magnitude and direction of the electric field can be more precisely and quickly changed than most other guidance cues such as chemical cues. Application of electric fields thus offers a robust experimental system for study of directional cell migration with extensive flexibility. We present a brief review of the background and describe the experimental system for studying electrotaxis.

Ellagic Acid Promotes Abeta42 Fibrillization and Inhibits Abeta42-induced Neurotoxicity

Smaller, soluble oligomers of beta-amyloid (Abeta) play a critical role in the pathogenesis of Alzheimer's disease (AD). Selective inhibition of Abeta oligomer formation provides an optimum target for AD therapy. Some polyphenols have potent anti-amyloidogenic activities and protect against Abeta neurotoxicity. Here, we tested the effects of ellagic acid (EA), a polyphenolic compound, on Abeta42 aggregation and neurotoxicity in vitro. EA promoted Abeta fibril formation and significant oligomer loss, contrary to previous results that polyphenols inhibited Abeta aggregation. The results of transmission electron microscopy (TEM) and Western blot displayed more fibrils in Abeta42 samples co-incubated with EA in earlier phases of aggregation. Consistent with the hypothesis that plaque formation may represent a protective mechanism in which the body sequesters toxic Abeta aggregates to render them harmless, our MTT results showed that EA could significantly reduce Abeta42-induced neurotoxicity toward SH-SY5Y cells. Taken together, our results suggest that EA, an active ingredient in many fruits and nuts, may have therapeutic potential in AD.

Reconstruction of Cicatricial Microstomia and Lower Facial Deformity by Windowed, Bipedicled Deep Inferior Epigastric Perforator Flap

We performed simultaneous facial scar repair and oral aperture open with a windowed, bilateral, bipedicled deep inferior epigastric perforator flap (DIEP flap) in a 20-year-old male patient who had suffered from severe postburn scar of the face and neck in association with serious cicatricial microstomia. DIEP flap is a typical perforator flap that has less donor site morbidity because of a minimal sacrifice of muscles. Since bipedicled DIEP flap can provide the largest skin territory in the lower abdominal wall and ensure a sufficient blood supply to zone IV, it is very suitable for the repair of massive defects of the face and neck. From our challenging case, we learned that bilateral, bipedicled DIEP flap is an excellent option for the repair of large faciocervical defects. Bilateral, bipedicled DIEP flap, which can produce an excellent esthetic and functional outcome, has reliable blood perfusion, provides soft and pliable tissue, and causes the minimal donor-site morbidity.

The Effect of Psychological Stress on Iron Absorption in Rats

Psychological stress (PS) is recognized as an important pathogenic factor which leads to metabolism disorder in many diseases. Previous studies have shown that systemic iron homeostasis in mammalians was changed under specific stress conditions.

Relationship Between HMGB1 Content and MHC-II Expression in Circulating Monocytes and Spleen of Mice Challenged with Zymosan

To observe the regularity of change in high mobility group protein box 1 (HMGB1) content in serum and spleen of mice with multiple organ dysfunction syndrome (MODS), to analyze the correlation between HMGB1 content and major histocompatibility complex (MHC)-II---I-A(b) expression on monocytes in blood and spleen, and to explore the effect of HMGB1 on immune function of circulating monocytes and splenocytes.

[Expression and Significance of CD147 and E-cadherin in Human Gastric Carcinoma]

To investigate the expression of CD147 and E-cadherin in gastric carcinoma and their correlation with clinicopathological features.

Human Liver Rate-limiting Enzymes Influence Metabolic Flux Via Branch Points and Inhibitors

Rate-limiting enzymes, because of their relatively low velocity, are believed to influence metabolic flux in pathways. To investigate their regulatory role in metabolic networks, we look at the global organization and interactions between rate-limiting enzymes and compounds such as branch point metabolites and enzyme inhibitors in human liver.

[Expression and Analysis of Recombinant Chicken IL-18 in Pichia Pastoris.]

Expression and analysis of recombinant chicken IL-18 in Pichia pastoris.

[Comparison of Different Definitions on Metabolic Syndrome in Obese Children.]

To compare the prevalence rates of metabolic syndrome (MS) in obese children, according to three commonly used 'Pediatric MS definitions': (1) the International Diabetes Federation (IDF), (2) Cook, et al, and(3)da Silva, et al, in order to choose an appropriate one for the Chinese obese children. It was also intended to assess the variances of American or Chinese cutoff values on MS prevalence.

4-Methoxy-anilinium Chloride

The crystal structure of the title compound, C(7)H(10)NO(+)·Cl(-), was synthesized by the reaction of 4-methoxy-aniline and hydro-chloric acid. In the crystal structure, the ions are involved in inter-molecular N-H⋯Cl hydrogen bonds.

1,1'-(2-Thienylmethylene)di-2-naphthol Ethyl Acetate Solvate

In the title compound, C(25)H(18)O(2)S·C(4)H(8)O(2), there are inter-molecular O-H⋯O hydrogen bonds between the main mol-ecule and the solvent molecule. The thio-phene ring is oriented at dihedral angles of 70.87 (7) and 75.36 (4)° with respect to the mean planes of the two naphthyl ring systems.

4-(6-Quinolyl-oxymeth-yl)benzonitrile

The title compound, C(17)H(12)N(2)O, was synthesized by an ether synthesis from quinolin-6-ol and 4-(bromo-meth-yl)benzonitrile. The phenyl ring of the benzonitrile group makes a dihedral angle of 47.52 (6)° with the plane of the quinoline fragment. The crystal structure is stabilized by inter-molecular C-H⋯π inter-actions between a benzene H atom of the benzonitrile group and the benzene ring of the quinoline fragment. In addition, the crystal structure also exhibits a weak inter-molecular C-H⋯N hydrogen bond.

6-(4-Nitro-benz-yloxy)quinoline

In the mol-ecule of the title compound, C(16)H(12)N(2)O(3), the nitrobenzene benzene ring forms a dihedral angle of 23.8 (8)° with the plane of the quinoline ring system. The crystal structure is stabilized by an aromatic π-π stacking inter-action between centrosymmetrically related benzene rings [centroid-centroid distance 3.663 (2) Å].

Three New Phenolic Glycosides and a New Triterpenoid from the Stems of Scolopia Chinensis

Three new phenolic glycosides, scolochinenosides C-E ( 1- 3), and a new triterpenoid, scolopianate A ( 4), were isolated from the stems of SCOLOPIA CHINENSIS, along with 15 known compounds. The structures of the new compounds were elucidated by means of extensive spectroscopic and chemical methods. Compound 3 contains a novel highly oxygenated lactone bridge ring attached at the aglycone. Six lanostane triterpenoids ( 10- 15) were discovered for the first time in a species other than GANODERMA LUCIDUM (Polyporaceae). In addition, the phenolic glycosides were evaluated for their inhibitory activity against snake venom phosphodiesterase I.

Endothelial Morphometry by Image Analysis of Corneas Organ Cultured at 31 Degrees C

To determine the factors influencing endothelial morphometry by using image analysis of corneas stored in organ culture to determine the coefficient of variation (CV) in cell area and percentage of hexagonal cells.

Impacts of Urban Forests on Offsetting Carbon Emissions from Industrial Energy Use in Hangzhou, China

This study quantified carbon storage and sequestration by urban forests and carbon emissions from energy consumption by several industrial sources in Hangzhou, China. Carbon (C) storage and sequestration were quantified using urban forest inventory data and by applying volume-derived biomass equations and other models relating net primary productivity (NPP) and mean annual biomass increments. Industrial energy use C emissions were estimated by accounting for fossil fuel use and assigning C emission factors. Total C storage by Hangzhou's urban forests was estimated at 11.74 Tg C, and C storage per hectare was 30.25 t C. Carbon sequestration by urban forests was 1,328, 166.55 t C/year, and C sequestration per ha was 1.66 t C/ha/year. Carbon emissions from industrial energy use in Hangzhou were 7 Tg C/year. Urban forests, through sequestration, annually offset 18.57% of the amount of carbon emitted by industrial enterprises, and store an amount of C equivalent to 1.75 times the amount of annual C emitted by industrial energy uses within the city. Management practices for improving Hangzhou's urban forests function of offsetting C emissions from energy consumption are explored. These results can be used to evaluate the urban forests' role in reducing atmospheric carbon dioxide.

Electric Currents and Lens Regeneration in the Rat

We studied the process of lens regeneration in the rat following an extracapsular lens extraction preserving the anterior lens capsule and anterior lens epithelium. We assessed clinically the clarity of the newly regenerated lens, evaluated changes in the lens electrical currents following surgery and during the regeneration process and correlated these changes with findings on light microscopy. Protein analysis of the regenerated lens was also undertaken. Experiments were performed in 41 Sprague-Dawley rats, sacrificed at 0, 2, 4 and 8 weeks postoperatively. Our results showed that complete lens regeneration occurred 8 weeks postoperatively only if the anterior epithelium was preserved and the lens capsule was closed surgically. Lens electrical currents, altered following surgery, recovered in parallel with the process of regeneration of the lens. The newly regenerated lens was optically clear and biochemical analysis revealed a pattern of protein expression resembling that observed during lens development. In conclusion, complete lens regeneration occurs in the rat and it is possible that lens electrical signals, together with other cues, may play an important role in this process.

The Associations of HA3G and HA3B MRNA Levels with HIV Disease Progression Among HIV-infected Individuals of China

To explore correlations between mRNA (hA3G, hA3F, and hA3B) levels and CD4 T-cell counts and HIV-1 viral loads to evaluate their respective roles in disease progression.

Aqueous Humor Dynamics During the Day and Night in Juvenile and Adult Rabbits

To determine the day and night differences in intraocular pressure (IOP), aqueous flow, outflow facility, uveoscleral outflow, and central corneal thickness (CCT) in juvenile and adult rabbits.

Regulation of Tissue Repair and Regeneration by Electric Fields

Endogenous electric fields (EFs) have been detected at wounds and damaged tissues. The potential roles of EFs in tissue repair and regeneration have been an intriguing topic for centuries. Recent researches have provided significant insights into how naturally occurring EFs may participate in the control of tissue repair and regeneration. Applied EFs equivalent to the size of fields measured in vivo direct cell migration, cell proliferation and nerve sprouting at wounds. More remarkably, physiological EFs are a guidance cue that directs cell migration which overrides other well accepted directional signals including initial injury stimulation, wound void, contact inhibition release, population pressure and chemotaxis. EFs activate many intracellular signaling pathways in a directional manner. Modulation of endogenous wound EFs affects epithelial cell migration, cell proliferation, and nerve growth at cornea wounds in vivo. Electric stimulation is being tested clinically for the treatments of bone fracture, wound healing and spinal cord injury. EFs thus may represent a novel type of signaling paradigm in tissue repair and regeneration. Combination of the electric stimulation and other well understood biochemical regulatory mechanisms may offer powerful and effective therapies for tissue repair and regeneration. This review introduces experimental evidence for the existence of endogenous EFs and discusses their roles in tissue repair and regeneration.

Long-term Outcome of Percutaneous Catheter Intervention for De Novo Coronary Bifurcation Lesions with Drug-eluting Stents or Bare-metal Stents

The purpose of this study was to assess the long-term risks and benefits of drug-eluting stents (DESs) compared with bare-metal stents (BMSs) for treatment of coronary bifurcation lesions.

Role of Heat-shock Protein 90 in Hepatitis E Virus Capsid Trafficking

p239 is a virus-like particle constituted from hepatitis E virus (HEV) recombinant proteins. It can be used as a surrogate for HEV and as an investigative tool to study cellular interactions because of its ability to adsorb to and penetrate HepG2 cellular membranes. Our objective was to use p239 to define the role of HEV capsid proteins during the early stages of infection. Pull-down and MALDI-TOF MS experiments identified three host-cell proteins, Grp 78/Bip, alpha-tubulin and heat-shock protein 90 (HSP90), and the latter was investigated further. Antibodies to p239 alone or HSP90 alone could detect p239 or HSP90, suggesting the formation of a complex between p239 and HSP90. In the HepG2 cell, geldanamycin (GA), an HSP90-specific inhibitor, blocked intracellular transportation of p239, but had no effect on the binding and cellular entry of p239, suggesting that HSP90 was important for HEV capsid intracellular transportation. RT-PCR results showed that the efficiency of wild-type HEV infection was inhibited significantly by GA treatment, suggesting the importance of HSP90 in virus infectivity. It was concluded that HSP90 plays a crucial role in the intracellular transportation of viral capsids in the early stage of HEV infection.

Chloride Channels and Transporters in Human Corneal Epithelium

Transport of water and electrolytes is critical for corneal clarity. Recent studies indicate another important function of transport of ions and electrolytes - establishing wound electric fields that guide cell migration. We found chloride (Cl(-)) flux is a major component of the corneal wound electric current. In order to elucidate the mechanisms of Cl(-) transport, we studied Cl(-) channels and transporters in human corneal epithelial (HCE) cells. We tested a transformed human corneal epithelial cell line (tHCE), primary cultures of human corneal epithelial cells (pHCE), and human donor corneas. We first used RT-PCR to determine expression levels of mRNA of CLC (Cl(-) channels/transporters of CLC gene family) family members and CFTR (cystic fibrosis transmembrane conductance regulator) in HCE cells. We then confirmed protein expression and distribution of selected CLC family members and CFTR with Western blot and immunofluorescence confocal microscopy. Finally, Cl(-) currents were recorded with electrophysiological techniques. The mRNAs of CLC-2, CLC-3, CLC-4, CLC-5, CLC-6, and CFTR were detected in the HCE cell line. CLC-1 and CLC-7 were not detectable. Western blot and immunostaining confirmed protein expression and distribution of CLC-2, CLC-3, CLC-4, CLC-6 and CFTR in human corneal epithelium. CLC-2 preferentially labeled the apical and basal layers, while CLC-3 and CLC-4 labeled only the superficial layer. CLC-6 and CFTR labeling showed a unique gradient with strong staining in apical layers which gradually decreased towards the basal layers. Corneal endothelium was positive for CLC-2, CLC-3, CLC-4, CLC-6 and possibly CFTR. Human corneal epithelial cells demonstrated voltage dependent Cl(-) currents. HCE cells express functional Cl(-) channels and transporters. CLC-2, CLC-3, CLC-4, CLC-6, and CFTR had distinct expression patterns in human corneal epithelium. Those molecules and their distribution may play important roles in maintaining resting Cl(-) fluxes and in regulating Cl(-) flux at corneal wounds, which may be a major contributor to wound electrical signaling.

Protective Effect of Puerarin on Acute Alcoholic Liver Injury

To provide experimental and theoretical basis for the clinical application of Puerarin in acute alcohol poisoning, 30 Wistar rats were randomized into 3 groups as follows: (1) Group A (control) underwent normal sodium (N.S.) peritoneal injection (i.p.) and intragastric administration (i.g.); (2) Group B (alcohol) underwent an equivalent dosage of N.S. i.p. and 40% ethanol (8000 mg/kg. d).ig for 5 days; (3) Group C (Puerarin) underwent Puerarin 200 mg/kg. d. ip, and an equivalent dosage of ethanol for 5 days. The left lobes of livers were sampled, and the levels of MDA, SOD and GPX in plasma and liver homogenate were detected. The level of MDA in plasma and liver homogenate in the alcohol group was obviously higher than that in the control group (p < 0.05, respectively), while that in the Puerarin group was significantly lower than in the alcohol group (p < 0.05, respectively). The levels of SOD and GPX were opposite to that of MDA. Under a light microscope, the livers of the rats in the alcohol group showed unclear structure of hepatic lobules, stiffness of hepatic sinusoids, diffused lipid degeneration of hepatic cells, cellular swelling, and focal necrosis, while the structure remained clear in the Puerarin group. Under the electron microscope, lipid degeneration, cell organ decrease, enlargement of endoplasmic reticulum, reduced quantity of hepatins and swelling of mitochondria were observed in cells of the model group. However, the pathologistic changes were slight in the Puerarin group. In conclusion, Puerarin may have the function of inhibiting the oxidative stress induced by acute alcoholism.

Effects of Physiological Electric Fields on Migration of Human Dermal Fibroblasts

Endogenous electric currents generated instantly at skin wounds direct migration of epithelial cells and are likely to be important in wound healing. Migration of fibroblasts is critical in wound healing. It remains unclear how wound electric fields guide migration of dermal fibroblasts. We report here that mouse skin wounds generated endogenous electric currents for many hours. Human dermal fibroblasts of both primary and cell-line cultures migrated directionally but slowly toward the anode in an electric field of 50-100 mV mm(-1). This is different from keratinocytes, which migrate quickly to the cathode. It took more than 1 hour for dermal fibroblasts to manifest detectable directional migration. Larger field strength (400 mV mm(-1)) was required to induce directional migration within 1 hour after onset of the field. Phosphatidylinositol-3-OH kinase (PI3 kinase) mediates cathode-directed migration of keratinocytes. We tested the role of PI3 kinase in anode-directed migration of fibroblasts. An applied electric field activated PI3 kinase/Akt in dermal fibroblasts. Dermal fibroblasts from p110gamma (a PI3 kinase catalytic subunit) null mice showed significantly decreased directional migration. These results suggest that physiological electric fields may regulate motility of dermal fibroblasts and keratinocytes differently, albeit using similar PI3 kinase-dependent mechanisms.

Study of Paraoxonase-1 Function on Tissue Damage of Dichlorvos

To examine the protective efficacy of paraoxonase-1 (PON1) against tissue damage caused by dichlorvos, purified rabbit PON1 was injected intravenously into rats 30min before they were given dichlorvos, while dichlorvos administration group and corn coil administration group were conducted to compare. Blood was collected at different time points after dichlorvos administration to examine the acetyl cholinesterase (AChE) inhibition level and clinical signs were observed after poisoning. 72h later, animals were anesthetized and the hippocampus, liver, lung and kidney were removed for observation of ultrastructure. AChE activities in PON1 pretreament group were statistically significant from dichlorvos administration group (P<0.01). The clinical signs were alleviated by PON1 significantly (P<0.05). The most common change of organophosphorus poisoning damage to liver was small lipid-like structures could be seen throughout the liver structure. In kidney, dense bodies were seen. The most significant changes in lung were lost of lamellar structure of lamellar bodies in type II alveolar epithelial cell. As for changes of hippocampus, demyaliation takes place after acute organophosphorus, but neural edema was not improved significantly in our study. In conclusion, PON1 can decrease the AChE inhibition, and alleviated clinical signs and tissue damage caused by dichlorvos.

Electrical Estimulation of Retinal Pigment Epithelial Cells

We investigated and characterized the effect of externally applied electric fields (EF) on retinal pigment epithelial (RPE) cells by exposing primary cultures of human RPE cells (hRPE) and those from the ARPE19 immortalized cell line to various strengths of EF (EF-treated cells) or to no EF (control cells) under different conditions including presence or absence of serum and gelatin and following wounding. We evaluated changes in RPE cell behavior in response to EF by using a computer based image capture and analysis system (Metamorph). We found that RPE cells responded to externally applied EFs by preferential orientation perpendicular to the EF vector, directed migration towards the anode, and faster translocation rate than control, untreated cells. These responses were voltage-dependent. Responses were observed even at low voltages, of 50-300 mV. Furthermore, the migration of hRPE cell sheets generated by wounding of confluent monolayers of cells at early and late confluence could be manipulated by the application of EF, with directed migration towards the anode observed at both sides of the wounded hRPE. In conclusion, RPE cell behaviour can be controlled by an externally applied EF. The potential for externally applied EF to be used as a therapeutic strategy in the management of selected retinal diseases warrants further investigation.

The Neuroretina is a Novel Mineralocorticoid Target: Aldosterone Up-regulates Ion and Water Channels in Müller Glial Cells

Glucocorticoids reduce diabetic macular edema, but the mechanisms underlying glucocorticoid effects are imperfectly elucidated. Glucocorticoids may bind to glucocorticoid (GR) and mineralocorticoid (MR) receptors. We hypothesize that MR activation may influence retinal hydration. The effect of the MR agonist aldosterone (24 h) on ion/water channel expression (real-time PCR, Western blot, immunofluorescence) was investigated on cultured retinal Müller glial cells (RMGs, which contribute to fluid homeostasis in the retina), in Lewis rat retinal explants, and in retinas from aldosterone-injected eyes. We evidenced cell-specific expression of MR, GR, and 11-beta-hydroxysteroid dehydrogenase type II. Aldosterone significantly enhances expression of sodium and potassium channels ENaC-alpha (6.5-fold) and Kir4.1 (1.9-fold) through MR and GR occupancy, whereas aquaporin 4 (AQP4, 2.9-fold) up-regulation is MR-selective. Aldosterone intravitreous injection induces retinal swelling (24% increase compared to sham-injected eyes) and activation of RMGs. It promotes additional localization of Kir4.1 and AQP4 toward apical microvilli of RMGs. Our results highlight the mineralocorticoid-sensitivity of the neuroretina and show that aldosterone controls hydration of the healthy retina through regulation of ion/water channels expression in RMGs. These results provide a rationale for future investigations of abnormal MR signaling in the pathological retina.

Evaluation of the Chinese Versions of the Minnesota Nicotine Withdrawal Scale and the Questionnaire on Smoking Urges-brief

To evaluate the reliability and validity of the Chinese versions of Minnesota Nicotine Withdrawal Scale and Questionnaire on Smoking Urges-Brief (MNWS-C and QSU-Brief-C) in Chinese smokers.

Single Amino Acid Changes in the Predicted RNase H Domain of Escherichia Coli RNase G Lead to Complementation of RNase E Deletion Mutants

The endoribonuclease RNase E of Escherichia coli is an essential enzyme that plays a major role in all aspects of RNA metabolism. In contrast, its paralog, RNase G, seems to have more limited functions. It is involved in the maturation of the 5' terminus of 16S rRNA, the processing of a few tRNAs, and the initiation of decay of a limited number of mRNAs but is not required for cell viability and cannot substitute for RNase E under normal physiological conditions. Here we show that neither the native nor N-terminal extended form of RNase G can restore the growth defect associated with either the rne-1 or rneDelta1018 alleles even when expressed at very high protein levels. In contrast, two distinct spontaneously derived single amino acid substitutions within the predicted RNase H domain of RNase G, generating the rng-219 and rng-248 alleles, result in complementation of the growth defect associated with various RNase E mutants, suggesting that this region of the two proteins may help distinguish their in vivo biological activities. Analysis of rneDelta1018/rng-219 and rneDelta1018/rng-248 double mutants has provided interesting insights into the distinct roles of RNase E and RNase G in mRNA decay and tRNA processing.

[Study on HPLC-FPS of Raw and Processed Fructus Polygoni Orientalis]

To establish the HPLC fingerprint method of Fructus Polygoni Orientalis before and after processed by choosing taxifolin as reference to compare the changes of chemical composition.

Enterobacter Spp.: a New Evidence Causing Bacterial Wilt on Mulberry

Thirty-six pathogenetic bacterial strains were isolated from wilted mulberry plants in Hangzhou, Zhejiang province of China. The six representative strains were confirmed to be involved in more than one Enterobacter species by common bacteriological test, electron microscope observation, hypersensitive reaction, Koch's postulates, physiological and biochemical test, biology, fatty acid methyl esters analysis (FAMEs), enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), 16s rRNA sequences analysis, and comparative analysis with 7 type strains and 3 reference strains. This is the first report on mulberry disease caused by Enterobacter spp. in the world providing new evidence on induction of the plant disease in this genus. The results are not only important in the mulberry disease management but also have significant scientific value for further studies of opportunistic human pathogens and environmental strains in Enterobacter.

Propane-1,3-diaminium Tetrachloridozincate(II) 18-crown-6 Clathrate

The reaction of propane-1,3-diamine hydrochloride, 18-crown-6 and zinc(II) chloride in methanol solution yields the title complex salt [systematic name: propane-1,3-diaminium tetrachloridozincate(II)-1,4,7,10,13,16-hexaoxacyclooctadecane (1/1)], (C(3)H(12)N(2))[ZnCl(4)] x C(12)H(24)O(6), with an unusual supramolecular structure. The diprotonated propane-1,3-diaminium cation forms an unexpected 1:1 supramolecular rotator-stator complex with the crown ether, viz. [C(3)H(12)N(2)(18-crown-6)](2+), in which one of the -NH(3)(+) substituents nests in the crown and interacts through N-H...O hydrogen bonding. The other -NH(3)(+) group interacts with the [ZnCl(4)](2-) anion via N-H...Cl hydrogen bonding, forming cation-crown-anion ribbons parallel to [010].

[Identification of Mentality Facticity Based on Multi-channel Event-related Potentials]

This paper explores the use of multi-channel event-related potentials (ERP) to identify mentality facticity or detect lie. Some identifiably meaning information, such as subjects' name and birthday, were selected as concealed information to be identified, 15 subjects were tested by concealed information test (CIT) paradigm and their electroencephalographs (EEG) were recorded from 30 electrodes. In virtue of analysis on the statistically significant difference between multi-channel ERPs evoked by probe information and that evoked by irrelevant information, the P300 amplitudes of 15 electrodes were selected as F-test samples. The significant difference of feature sample values between probe and irrelevant information was applied to identify mentality facticity. The results indicate that P300 amplitudes evoked in many electrodes are significantly different between probe and irrelevant information (P < 0.01). According to the significant difference of space sample values between probe and irrelevant information, the detection correctness to probe information reaches to 93.3% and is better than that of methods based single-channel ERP. The method proposed has the advantages of non-invasion and better accuracy, which could be used to identify mentality facticity effectively.

Bis(adamantan-1-aminium) Tetrachloridozincate(II) 18-crown-6 Monohydrate Clathrate

The structure of the title compound [systematic name: bis(adamantan-1-aminium) tetrachloridozincate(II)-1,4,7,10,13,16-hexaoxacyclooctadecane-water (1/1/1)], (C(10)H(18)N)(2)[ZnCl(4)] x C(12)H(24)O(6) x H(2)O, consists of supramolecular rotator-stator assemblies and ribbons of hydrogen bonds parallel to [010]. The assemblies are composed of one protonated adamantan-1-aminium cation and one crown ether molecule (1,4,7,10,13,16-hexaoxacyclooctadecane) to give an overall [(C(10)H(18)N)(18-crown-6)](+) cation. The -NH(3)(+) group of the cation nests in the crown and links to the crown-ether O atoms through N-H...O hydrogen bonds. The 18-crown-6 ring adopts a pseudo-C(3v) conformation. The second adamantan-1-aminium forms part of ribbons of adamantan-1-aminium-water-tetrachloridozincate units which are interconnected by O-H...Cl, N-H...O and N-H...Cl hydrogen bonds via three different continuous rings with R(5)(4)(12), R(4)(3)(10) and R(3)(3)(8) motifs.

BNP7787-mediated Modulation of Paclitaxel- and Cisplatin-induced Aberrant Microtubule Protein Polymerization in Vitro

Taxane and platinum drugs are important agents in the treatment of cancer and have shown activity against a variety of tumors, including ovarian, breast, and lung cancer, either as single agents or in combination with other chemotherapy drugs. However, a serious and prevalent side effect of taxane (docetaxel and all formulations/derivatives of paclitaxel) and platinum (cisplatin, carboplatin, and oxaliplatin) agents is dose-limiting chemotherapy-induced peripheral neuropathy (CIPN). CIPN can result in treatment delays, dose modifications, and, in severe cases, discontinuation of chemotherapy. Consequently, effective treatments for CIPN are needed. Dimesna (BNP7787; Tavocept; disodium 2,2'-dithio-bis-ethanesulfonate) is an investigational drug that is undergoing international clinical development as a treatment that is coadministered with first-line taxane and platinum combination chemotherapy in patients with inoperable advanced primary adenocarcinoma of the lung. BNP7787 is currently being developed with the objective of increasing the survival of cancer patients receiving taxane- and/or cisplatin-based chemotherapy. Additional data indicate that BNP7787 may also protect against common and serious chemotherapy-induced toxicities, including chemotherapy-induced anemia, nausea, emesis, nephrotoxicity, and neuropathy, without interfering with antitumor activity of the chemotherapeutic agent(s). Studies herein show that BNP7787 prevents aberrant microtubule protein (MTP) polymerization that is caused by exposure of MTP to paclitaxel or cisplatin. BNP7787 modulates paclitaxel-induced hyperpolymerization of MTP in a dose-dependent manner, and mesna, an in vivo metabolite of BNP7787, protects against time-dependent cisplatin-induced inactivation of MTP. We propose that interactions between BNP7787 and MTP may play a role in BNP7787-mediated protection against CIPN.

Auto-mobilized Adult Hematopoietic Stem Cells Advance Neovasculature in Diabetic Retinopathy of Mice

Hematopoietic stem cells (HSCs) can be used to deliver functionally active angiostatic molecules to the retinal vasculature by targeting active astrocytes and may be useful in targeting pre-angiogenic retinal lesions. We sought to determine whether HSC mobilization can ameliorate early diabetic retinopathy in mice.

[Chemical Constituents of Discocleidion Rufescens]

To study chemical constituents of leaves from Discocleidion rufescens.

Multipotent Adipose Stromal Cells and Breast Cancer Development: Think Globally, Act Locally

It has long been appreciated that stromal cells within the breast tumor microenvironment contribute to mammary carcinogenesis. However, to date, very little is known regarding the role of local adipose-derived stromal cells (ASCs) in the development of breast cancer. Based on pathological, epidemiological and experimental data, we postulate that breast-derived ASCs are unique mesenchymal stem-like cells that play a critical role in the development of breast cancer and discuss the global implications of this working model in terms of breast cancer prevention, early detection, and new targeted therapies.

Diverse Ecdysterones Show Different Effects on Amyloid-β42 Aggregation but All Uniformly Inhibit Amyloid-β42-induced Cytotoxicity

Amyloid-β (Aβ) plays a pivotal role in Alzheimer's disease (AD) pathogenesis and in toxic mechanisms such as oxidative stress, mitochondrial dysfunction, calcium turbulence, and apoptosis induction. Therefore, interfering with Aβ aggregation has long been one of the most promising strategies for AD treatment. Ecdysterones (ECRs) are steroidal hormones in insects and terrestrial plants that have high structural diversity and multiple beneficial pharmacological activities. Here, we studied the effects of six ECRs on Aβ aggregation and cytotoxicity. Two ECRs with an acetoxyl group at the 2 or 3 position and saturated chains as side groups showed apparent promotion of Aβ42 fibrilization, resulting in less Aβ42 oligomers in the samples. Another three with unsaturated side chains clearly inhibited Aβ aggregation and disaggregated preformed fibrils, but increased the Aβ42 oligomer levels. Nevertheless, our MTT results showed that all ECRs tested inhibited Aβ42-induced cytotoxicity. This protective activity may be partly attributable to ECR-mediated amelioration of A&beta42-induced release of reactive oxygen species. Taken together, our findings suggest that ECRs, a series of natural compounds in many plants and insects, have therapeutic potential in AD and that the deduced structure-activity relationships may be beneficial in drug design for the treatment of AD and other amyloidoses.

[Efficiency of Human and Rat Bone Marrow Mesenchymal Stem Cells Transfected by Modified Adenovirus]

To compare the efficiency of human bone marrow mesenchymal stem cells (hBMSCs) with rat BMSCs (rBMSCs) transfected by modified adenovirus containing fiber 35 (AdF35)-enhanced green fluorescence protein(eGFP).

[Research Advances in Denitrogenation Characteristics of Aerobic Denitrifiers]

The discovery of aerobic denitrifiers is the enrichment and breakthrough of traditional denitrification theory. Owing to their unique superiority in denitrogenation, aerobic denitrifiers have become a hotspot in the study of bio-denitrogenation of waste water. Under aerobic conditions, the aerobic denitrifiers can utilize organic carbon sources for their growth, and produce N2 from nitrate and nitrite. Most of the denitrifiers can also proceed with heterotrophic nitrification simultaneously, transforming NH4(+)-N to gaseous nitrogen. In this paper, the denitrogenation characteristics and action mechanisms of some isolated aerobic denitrifiers were discussed from the aspects of electron theory and denitrifying enzyme system. The effects of the environmental factors DO, carbon sources, and C/N on the denitrogenation process of aerobic denitrifiers were analyzed, and the screening methods as well as the present and potential applications of aerobic denitrifiers in wastewater treatment were described and discussed.

A Time-lapse and Quantitative Modelling Analysis of Neural Stem Cell Motion in the Absence of Directional Cues and in Electric Fields

Neural stem cell (NSC) migration is an important component of their developmental function and therapeutic potential. Understanding their mode of migration and their response to guidance cues can contribute to improved therapies for CNS repair, in which appropriate homing to sites of injury is essential. Using time-lapse imaging, we have analyzed the NSC mode of migration in vitro, both in the absence of directional cues and in the presence of applied electric fields (EFs), previously shown to constitute a strong directional signal for these cells. Without EFs, NSCs displayed an amoeboid motion, characterized by small lamellipodial-like protrusions with changing orientations, leading to highly tortuous migration. In EFs, tortuosity diminished as electrotaxis toward the cathode occurred. EFs suppressed the formation of protrusions oriented toward the anode, suggesting that restriction of protrusions with opposing orientation could underlie the change from tortuous motion to directed migration. Treatment with LY294002, a phosphatidylinositol-3-OH kinase (Pi3K) inhibitor, reduced the cathodal bias of protrusions in EFs and the frequency of changes in direction. We generated a model of NSC migration with only two key parameters, which could accurately reproduce experimental migration patterns, and we used it to show that both effects of LY294002 contribute to impair electrotaxis, although decreased protrusion bias is the most important. Our results show that control of protrusion orientation by EFs is an important component of the electrotactic response. A simple modelling approach might be useful in understanding how diverse pharmacological treatments or genetic deletions affect different kinds of directional cell migration.

Coupling of Receptor Conformation and Ligand Orientation Determine Graded Activity

Small molecules stabilize specific protein conformations from a larger ensemble, enabling molecular switches that control diverse cellular functions. We show here that the converse also holds true: the conformational state of the estrogen receptor can direct distinct orientations of the bound ligand. 'Gain-of-allostery' mutations that mimic the effects of ligand in driving protein conformation allowed crystallization of the partial agonist ligand WAY-169916 with both the canonical active and inactive conformations of the estrogen receptor. The intermediate transcriptional activity induced by WAY-169916 is associated with the ligand binding differently to the active and inactive conformations of the receptor. Analyses of a series of chemical derivatives demonstrated that altering the ensemble of ligand binding orientations changes signaling output. The coupling of different ligand binding orientations to distinct active and inactive protein conformations defines a new mechanism for titrating allosteric signaling activity.

α-Tocopherol Quinone Inhibits β-amyloid Aggregation and Cytotoxicity, Disaggregates Preformed Fibrils and Decreases the Production of Reactive Oxygen Species, NO and Inflammatory Cytokines

Alzheimer's disease (AD) is a complex, multifactorial neurodegenerative disease. The aggregation of beta-amyloid (Aβ) into extracellular fibrillar deposition is a pathological hallmark of AD. The Aβ aggregate-induced neurotoxicity, inflammatory reactions and oxidative stress are linked strongly to the etiology of AD. The currently available hitting-one-target drugs are insufficient for the treatment of AD. Therefore, finding multipotent agents able to modulate multiple targets simultaneously is attracting more attention. Previous studies indicated that vitamin E or its constituent such as α-tocopherol (α-T) was able to attenuate the effects of several pathogenetic factors in AD. However, ineffective or detrimental results were obtained from a number of clinical trials of vitamin E. Here, we showed that naturally synthesized RRR-α-tocopherol quinone (α-TQ), a main derivative of α-T, could inhibit Aβ42 fibril formation dose-dependently. Further investigations indicated that α-TQ could attenuate Aβ42-induced neurotoxicity toward SH-SY5Y neuroblastoma cells, disaggregate preformed fibrils and interfere with natural intracellular Aβ oligomer formation. Moreover, α-TQ could decrease the formation of reactive oxygen species (ROS) and NO, and modulate the production of cytokines by decreasing TNF-α and IL-1β and increasing IL-4 formation in microglia. Taken together, α-TQ targeting multiple pathogenetic factors deserves further investigation for prevention and treatment of AD.

Preferential Loss of Th17 Cells is Associated with CD4 T Cell Activation in Patients with 2009 Pandemic H1N1 Swine-origin Influenza A Infection

Very limited evidence has been reported on host T cell responses to the pandemic H1N1 swine-origin influenza A virus (S-OIV) infection in humans. Therefore, we investigated the proportions of peripheral T cell subsets and analyzed the relationship of T helper subset changes with T cell activation during this infection. We found that these S-OIV-infected patients exhibited rapid lymphopenia, T cell activation and preferential loss of Th17 subset at the early stage of acute infection. Statistical analysis indicated that CD4 depletion and loss of Th17 cells, rather than Th1 or Treg cells, were correlated with CD4 T cell activation. More importantly, up-regulated IFN-α likely contributed to the functional loss of Th17 cells. Thus, rapidly generalized lymphopenia, preferential loss of Th17 population and T cell activation presented as characteristics of the early immune response in S-OIV-infected patients. These findings, therefore, may be helpful for an earlier diagnosis and further studies of immune pathogenesis of S-OIV infection.

SB203580, a P38 Mitogen-activated Protein Kinase Inhibitor, Suppresses the Development of Endometriosis by Down-regulating Proinflammatory Cytokines and Proteolytic Factors in a Mouse Model

p38 mitogen-activated protein kinase (p38 MAPK), a regulator of inflammation, may play a role in the pathogenesis of endometriosis (EM). We studied the effect of SB203580, a p38 MAPK inhibitor, on the development of EM in a mouse model.

[A Case Report of Monitoring on Carbamazepine in Breast Feeding Woman]

To evaluate the safety of oral cabamazepine during breast milk feeding. The carbamazepine concentration in breast milk of one epilepsy maternal patient was assayed by high performance liquid chromatography, and the literature was reviewed to find the nursing evidence in the use of cabamazepine. The carbamazepine concentration in breast milk ranged from 0.34-0.86 mg/L. The neonate daily dose intake was estimated ranging from 0.34 mg to 0.86 mg through breast-feeding in theory. The literature showed that carbamazepine was generally considered safe for use during breast feeding; however, adverse effects should be monitored as recommended. It is better to avoid feeding at high concentration level to minimize the harm of carbamazepine to the baby.

[Clinical and Laboratory Characteristics of a New Influenza A (HIN1) Epidemic]

To analysis the clinical and laboratory characteristics of Patients infected with new influenza A (HIN1) virus.

PathLocdb: a Comprehensive Database for the Subcellular Localization of Metabolic Pathways and Its Application to Multiple Localization Analysis

In eukaryotes, the cell is divided into several compartments enclosed by unitary membranes. Such compartmentalization is critical for cells to restrict different pathways to be carried out in different subcellular regions. The summary and classification of subcellular localizations of metabolic pathways are the first steps towards understanding their roles in spatial differentiation and the specialization of metabolic pathways in different organisms.

[Study on Excretion of Stilbene Glycoside (THSG) and Its Beta-cyclodextrin Inclusion]

The excretion characteristics of stilbene glycoside (THSG) and its beta-cyclodextrin inclusion in bile, urine and feces after oral administration to rats were studied. Bile for 24 h, urine and feces for 72 h were collected. The content of THSG was determined by HPLC-UV. The established HPLC-UV method was available for the analysis of THSG in excreta and corresponded to the requirement of biological sample analysis. After given THSG and its beta-cyclodextrin inclusion, the amount of prototype THSG in feces were 3.27% and 0.61%, meanwhile THSG in bile were 0.20% and 0.18%, respectively. Only a little THSG was found in urine. The result showed that beta-cyclodextrin inclusion reduced the fecal excretion of THSG. However, the characteristic of urinary and biliary excretion wasn't changed.

Patterns of Childhood Trauma and Psychological Distress Among Injecting Heroin Users in China

Childhood trauma has been reported as a possible cause of future substance abuse in some countries. This study reports the prevalence of childhood trauma and examines its association with psychological distress among injecting drug users from mainland China.

[Difference of Immunological and Inflammatory Indices Between Mild Type and Severe Type of Adult Pandemic (H1N1) Patients]

To study the difference of immunological and inflammatory indices between mild type and severe type of adult pandemic (H1N1) patients.

[Changes and Analysis of Peripheral White Blood Cells and Lymphocyte Subsets for Patients with Pandemic Influenza A Virus (H1N1) Infection]

To investigate the characters and changes of peripheral white blood cells and lymphocyte subsets of patients with pandemic influenza A virus (H1N1) infection and to provide evidences for diagnosis, treatment and prognosis of influenza A (H1N1) infection.

[Proteomics of Rice Leaf and Grain at Late Growth Stage Under Different Nitrogen Fertilization Levels]

Taking super-rice Liangyoupeijiu as test material, and by the method of two-dimensional gel electrophoresis (2-DE), this paper studied the changes in the leaf and grain proteomics of the variety at its late growth stage under different levels of nitrogen fertilization (1/2 times of normal nitrogen level, 20 mg x L(-1); normal nitrogen level, 40 mg x L(-1); 2 times of normal nitrogen level, 80 mg x L(-1)), with the biological functions of 16 leaf proteins, 9 inferior grain proteins, and 4 superior grain proteins identified and analyzed. Nitrogen fertilization could affect and regulate the plant photosynthesis via affecting the activation of photosynthesis-related enzymes and of CO2, the light system unit, and the constitution of electron transfer chain at the late growth stage of the variety. It could also promote the expression of the enzymes related to the energy synthesis and growth in inferior grains. High nitrogen fertilization level was not beneficial to the synthesis of starch in superior grain, but sufficient nitrogen supply was still important for the substance accumulation and metabolism. Therefore, rational nitrogen fertilization could increase the photosynthesis rate of flag leaves, enhance the source function, delay the functional early ageing, and promote the grain-filling at late growth stage.

6-Chloro-8-methyl-4H-3,1-benzoxazine-2,4(1H)-dione

The two mol-ecules in the asymmetric unit of the title compound, C(9)H(6)ClNO(3), are nearly planar, with r.m.s. deviations of 0.034 and 0.037 Å. The crystal structure is stabilized by two weak inter-molecular N-H⋯O inter-actions.

Bis(4-fluoro-anilinium) Tetra-chloridocuprate(II)

The crystal structure of the title compound, (C(6)H(7)FN)(2)[CuCl(4)], consists of parallel two-dimensional perovskite-type layers of corner-sharing CuCl(6) octa-hedra. These are bonded together via N-H⋯Cl hydrogen bonds from the 4-fluoro-anilinium chains, which are almost perpendicular to the layers. The CuCl(4) dianions have two short Cu-Cl bonds [2.2657 (15) and 2.2884 (13) Å] and two longer bonds [2.8868 (15) Å], giving highly Jahn-Teller-distorted CuCl(6) octa-hedra. The Cu atoms are situated on crystallographic centers of inversion.

1-[Morpholino(phen-yl)meth-yl]-2-naphthol

There are two independent mol-ecules in the asymmetric unit of the title compound, C(21)H(21)NO(2), which was synthesized by the one-pot reaction of 2-naphthol, morpholine and benzaldehyde. The dihedral angles between the naphthalene ring systems and the benzene rings are 84.03 (7) and 75.76 (8)° in the two mol-ecules and an intra-molecular O-H⋯N hydrogen bond occurs in each independent mol-ecule.

Melaminium Iodide Monohydrate

In the title melaminium salt, 2,4,6-triamino-1,3,5-triazin-1-ium iodide monohydrate, C(3)H(7)N(6) (+)·I(-)·H(2)O, the components are linked via N-H⋯O, N-H⋯N, O-H⋯I and N-H⋯I hydrogen bonds. All of the H atoms of the melaminium cation are involved in hydrogen bonds. The melaminium cations are inter-connected by four N-H⋯N hydrogen bonds, forming ribbons along [111]. The water mol-ecules connected by N-H⋯O hydrogen bonds also form part of these ribbons. The ribbons are inter-connected by other hydrogen bonds (O-H⋯I and N-H⋯I), as well as by π-π inter-actions [centroid-centroid distance = 3.6597 (17) Å].

Melaminium Perchlorate Monohydrate

In the title hydrated salt, 2,4,6-triamino-1,3,5-triazin-1-ium perchlorate monohydrate, C(3)H(7)N(6) (+)·ClO(4) (-)·H(2)O, the constituents are linked via hydrogen bonds of the O-H⋯O, N-H⋯O, N-H⋯N and N-H⋯Cl types. All the H atoms of the melaminium cation are involved in the hydrogen bonds. The melaminium residues are inter-connected by four N-H⋯N hydrogen bonds, forming chains parallel to (111). The ribbons are inter-connected by other hydrogen bonds as well as by π-π inter-actions [centroid-centroid distance = 3.8097 (7) Å].

Dimethyl-ammonium Tetra-chloridoferrate(III) 18-crown-6 Clathrate

The reaction of dimethyl-amine hydro-chloride, 18-crown-6 and ferric chloride in ethanol yields the title compound, (C(2)H(8)N)[FeCl(4)]·C(12)H(24)O(6), which exhibits an unusual supramolecular structure. The protonated dimethyl-amine contains one NH(2) (+) group, resulting in a 1:1 supra-molecular rotator-stator structure (CH(3)-NH(2) (+)-CH(3))(18-crown-6), through N-H⋯O hydrogen-bonding inter-actions between the ammonium group of the cation and the O atoms of the crown ether. In the crystal, all three components lie on a common crystallographic mirror plane normal to [010].

Benzyl-ammonium Tetra-fluoro-borate 18-crown-6 Clathrate

The reaction of benzyl-ammonium tetra-fluoro-borate and 18-crown-6 in a methano-lic solution yields the title compound, C(7)H(10)N(+)·BF(4) (-)·C(12)H(24)O(6)O6, which displays a supra-molecular structure. The -NH(3) (+) substituent of the benzyl-ammonium cation forms a 1:1 supra-molecular rotator-stator structure by N-H⋯O hydrogen-bonding inter-actions.

1-Cyano-methyl-1,4-diazo-niabicyclo-[2.2.2]octane Tetra-chloridomanganate(II)

In the crystal structure of the title compound, (C(8)H(15)N(3))[MnCl(4)], the Mn atom is coordinated by four chloride ligands in a slightly distorted tetra-hedral geometry. Each [MnCl(4)](2-) anion is connected to the 1-cyano-methyl-1,4-diazo-niabicyclo-[2.2.2]octane dications by N-H⋯Cl hydrogen bonds, forming chains parallel to [001].

4-Bromo-anilinium Perchlorate 18-crown-6 Clathrate

The reaction of 4-bromo-aniline, 18-crown-6, and perchloric acid in methanol yields the title compound, C(6)H(7)BrN(+)·ClO(4) (-)·C(12)H(24)O(6), in which the protonated -NH(3) (+) group forms three bifurcated N-H⋯O hydrogen bonds to the O atoms of the crown ether.

Electrical Activation of Wound-Healing Pathways

BACKGROUND: Effective wound healing has been a lasting and challenging topic in health care. Various strategies have been used to accelerate and perfect the healing process. One such strategy has involved the application of an exogenous electrical stimulus to chronic wounds with the aim of stimulating healing responses. THE PROBLEM: The biology of electric stimulation to instigate healing, however, is very poorly understood. How does electric stimulation induce healing responses? BASIC/CLINICAL SCIENCE ADVANCES: Recent research shows that the electric fields (EFs) activate multiple signaling pathways that are critical for wound healing. Importantly, the EFs provide a powerful, sometimes an overriding, directional signal for cell migration in wound healing. Unlike other stimuli, EFs have the intrinsic property of being directional. The EF-directed cell migration (electrotaxis/galvanotaxis) appears to be a consequence of EF-induced polarized signaling of epidermal growth factor receptors, integrins, and phosphoinositide 3 kinase/Pten, and may be mediated by protein kinase C, intracellular Ca(2+), and cyclic adenosine monophosphate (cAMP). Because directional cell migration is a key component in wound healing, galvanotaxis may represent an important mechanism of wound healing. CLINICAL CARE RELEVANCE: With the constantly enlarging diabetic and aging population, chronic or nonhealing wounds pose increasing health and economic problems, and currently there is no effective therapy available. Electric stimulation activates important intracellular signaling pathways that are polarized in the EF direction, resulting in enhanced and stimulated directional cell migration. Electric stimulation offers a novel approach to achieve better and accelerated wound healing. CONCLUSION: Experimental evidence suggests a significant role of endogenous EFs in cell migration in wound healing. Most importantly, EFs are a very powerful signal to direct cell migration. Electric stimulation therefore may represent a promising and unique strategy to induce cell and tissue growth in a directional manner, to enhance wound healing, and to achieve better wound healing.

Association of Variations of NAb 2F5 and 4E10 Epitopes and Disease Progression in Chinese Antiretroviral Treatment-naïve Patients Infected with HIV-1 Clade B'

Studies on human immunodeficiency virus type 1 (HIV-1) vaccines have recently focused on targeting the conserved neutralizing epitopes 2F5 and 4E10, and hence it is important to understand the extent of mutations in these two viral epitopes. Here, we investigated the amino acid mutations in epitopes of 2F5 (ELDKWA, HIV-1 HXB2 env 662 - 667 aa) and 4E10 (NWFDIT, HIV-1 HXB2 env 671 - 676 aa) in the membrane proximal-external region of gp41 from clade B' HIV-1-infected individuals living in Henan province, China. We also examined the frequency of a mutation and its relation to disease progression.

The Role of Endoplasmic Reticulum Stress in the Early Stage of Diabetic Retinopathy

The aim of this study was to evaluate the role of endoplasmic reticulum (ER) stress in diabetic retinopathy (DR) using in vitro and in vivo models. For the in vivo studies, diabetes was induced in rats, and retinal expression of glucose-regulated protein 78 (GRP78), activating transcription factor 4 (ATF4), C/EBP homologous protein (CHOP), and vascular endothelial growth factor (VEGF) in groups of rats at 1, 3, and 6 months was assessed. For the in vitro studies, human retinal capillary endothelial cells (HRCECs) were cultured in the presence of varying glucose concentrations, and the expression of mRNA and protein for GRP78, ATF4, CHOP, and VEGF was assessed. The chosen glucose concentrations were reflective of those apparent in diabetic patients. Expression of VEGF and CHOP mRNA levels were significantly increased in diabetic rats compared to control rats at 1, 3, and 6 months (P < 0.05). In HRCECs cultured in the presence of high as well as variable glucose concentrations, CHOP expression and apoptosis were significantly increased (P < 0.05). However, GRP78 and ATF4 expression levels were unchanged. Our findings suggest that early progression of DR may be mediated by ER stress, but probably does not involve changes in ATF4 or GRP78.

Effect of PON1 on Dichlorvos Toxicokinetics

To provide toxicokinetic and clinical evidence of the hydrolytic effect of paraoxonase-1 (PON1) on acute organophosphate poisoning in rats.

Bilobed Mucosal Flap for Correction of Secondary Lip Deformities Following Cleft Lip Surgery

Upper lip whistling deformities, asymmetric upper lip thickness and insufficient vermilion tubercle often occur simultaneously in an affected individual. Unfortunately, these deformities cannot be corrected by a single conventional method. Here, we describe a new technique, using a vermilion-bilobed flap, to address the triplex of deformities simultaneously.

Acute Carbamazepine Poisoning Treated with Resin Hemoperfusion Successfully

Carbamazepine (CBZ) poisoning has been occurring more frequently. We describe the use of synthesized resin-absorbed hemoperfusion in the therapy of a 48-year-old man who developed carotic, cardiovascular shock and multiorgan dysfunction due to a CBZ overdose (the highest concentration of drug >20 mg/L; therapeutic range, 8-12 mg/L). The treatment was very successful; and the patient eventually was discharged with a full recovery and no complications, although his diagnosis and treatment had been delayed for 56 hours. Hemoperfusion has a steady clearance of this drug without subsequent rebound or potential hazards. Resin hemoperfusion should be first considered for acute CBZ intoxication, especially when drug-induced gastrointestinal hypomotility prevents elimination via the gut and patient is under life-threatening condition.

The Role of Electrical Signals in Murine Corneal Wound Re-epithelialization

Ion flow from intact tissue into epithelial wound sites results in lateral electric currents that may represent a major driver of wound healing cell migration. Use of applied electric fields (EF) to promote wound healing is the basis of Medicare-approved electric stimulation therapy. This study investigated the roles for EFs in wound re-epithelialization, using the Pax6(+/-) mouse model of the human ocular surface abnormality aniridic keratopathy (in which wound healing and corneal epithelial cell migration are disrupted). Both wild-type (WT) and Pax6(+/-) corneal epithelial cells showed increased migration speeds in response to applied EFs in vitro. However, only Pax6(+/+) cells demonstrated consistent directional galvanotaxis towards the cathode, with activation of pSrc signaling, polarized to the leading edges of cells. In vivo, the epithelial wound site normally represents a cathode, but 43% of Pax6(+/-) corneas exhibited reversed endogenous wound-induced currents (the wound was an anode). These corneas healed at the same rate as WT. Surprisingly, epithelial migration did not correlate with direction or magnitude of endogenous currents for WT or mutant corneas. Furthermore, during healing in vivo, no polarization of pSrc was observed. We found little evidence that Src-dependent mechanisms of cell migration, observed in response to applied EFs in vitro, normally exist in vivo. It is concluded that endogenous EFs do not drive long-term directionality of sustained healing migration in this mouse corneal epithelial model. Ion flow from wounds may nevertheless represent an important component of wound signaling initiation.

Massive Digestive Tract Bleeding Due to Pancreatic Pseudocyst: a Case Report

PI3K Mediated Electrotaxis of Embryonic and Adult Neural Progenitor Cells in the Presence of Growth Factors

Correct guidance of the migration of neural progenitor cells (NPCs) is essential for the development and repair of the central nervous system (CNS). Electric field (EF)-guided migration, electrotaxis, has been observed in many cell types. We report here that, in applied EFs of physiological magnitude, embryonic and adult NPCs show marked electrotaxis, which is dependent on the PI3K/Akt pathway. The electrotaxis was also evidenced by ex vivo investigation that transplanted NPCs migrated directionally towards cathode in organotypic spinal cord slice model when treated with EFs. Genetic disruption or pharmacological inhibition of phosphoinositide 3-kinase (PI3K) impaired electrotaxis, whereas EF exposure increased Akt phosphorylation in a growth factor-dependent manner and increased phosphatidylinositol-3,4,5-trisphosphate (PIP3) levels. EF treatments also induced asymmetric redistribution of PIP3, growth factor receptors, and actin cytoskeleton. Electrotaxis in both embryonic and adult NPCs requires epidermal growth factor (EGF) and fibroblast growth factor (FGF). Our results demonstrate the importance of the PI3K/Akt pathway in directed migration of NPCs driven by EFs and growth factors and highlight the potential of EFs to enhance the guidance of various NPC populations in CNS repair therapies.

17beta-estradiol Attenuates Programmed Cell Death in Cortical Pericontusional Zone Following Traumatic Brain Injury Via Upregulation of ERalpha and Inhibition of Caspase-3 Activation

Pericontusional zone (PCZ) of traumatic cerebral contusion is a target of pharmacological intervention. It is well studied that 17beta-estradiol has a protective role in ischemic brain injury, but its role in brain protection of traumatic brain damage deserves further investigation, especially in pericontusional zone. Here we show that 17beta-estradiol enhances the protein expression and mRNA induction of estrogen alpha receptor (ERalpha) and prevents from programmed cell death in cortical pericontusional zone. ERalpha specific antagonist blocks this protective effect of 17beta-estradiol. Caspase-3 activation occurs in cortical pericontusional zone of the oil-treated injured rat brain and its activation is inhibited by 17beta-estradiol treatment. Additionally, ERalpha specific antagonist reverses this inhibition. Pan-caspase inhibitor also protect cortical pericontusional zone from programmed cell death. Our present study indicates 17beta-estradiol protects from programmed cell death in cortical pericontusional zone via enhancement of ERalpha and decrease of caspase-3 activation.

Modulating Endogenous Electric Currents in Human Corneal Wounds--a Novel Approach of Bioelectric Stimulation Without Electrodes

To measure electric current in human corneal wounds and test the feasibility of pharmacologically enhancing the current to promote corneal wound healing.

GSK-3β is Essential for Physiological Electric Field-directed Golgi Polarization and Optimal Electrotaxis

Endogenous electrical fields (EFs) at corneal and skin wounds send a powerful signal that directs cell migration during wound healing. This signal therefore may serve as a fundamental regulator directing cell polarization and migration. Very little is known of the intracellular and molecular mechanisms that mediate EF-induced cell polarization and migration. Here, we report that Chinese hamster ovary (CHO) cells show robust directional polarization and migration in a physiological EF (0.3-1 V/cm) in both dissociated cell culture and monolayer culture. An EF of 0.6 V/cm completely abolished cell migration into wounds in monolayer culture. An EF of higher strength (≥1 V/cm) is an overriding guidance cue for cell migration. Application of EF induced quick phosphorylation of glycogen synthase kinase 3β (GSK-3β) which reached a peak as early as 3 min in an EF. Inhibition of protein kinase C (PKC) significantly reduced EF-induced directedness of cell migration initially (in 1-2 h). Inhibition of GSK-3β completely abolished EF-induced GA polarization and significantly inhibited the directional cell migration, but at a later time (2-3 h in an EF). Those results suggest that GSK-3β is essential for physiological EF-induced Golgi apparatus (GA) polarization and optimal electrotactic cell migration.

Downregulation of PTEN at Corneal Wound Sites Accelerates Wound Healing Through Increased Cell Migration

The PI3K/Akt pathway is required for cell polarization and migration, whereas the phosphatase and tensin homologue deleted on chromosome 10 (PTEN) has inhibitory effects on the PI3K/Akt pathway. The authors therefore hypothesized that wounding would downregulate PTEN and that this downregulation would enhance wound healing.

TSdb: a Database of Transporter Substrates Linking Metabolic Pathways and Transporter Systems on a Genome Scale Via Their Shared Substrates

TSdb ( http://tsdb.cbi.pku.edu.cn ) is the first manually curated central repository that stores formatted information on the substrates of transporters. In total, 37608 transporters with 15075 substrates from 884 organisms were curated from UniProt functional annotation. A unique feature of TSdb is that all the substrates are mapped to identifiers from the KEGG Ligand compound database. Thus, TSdb links current metabolic pathway schema with compound transporter systems via the shared compounds in the pathways. Furthermore, all the transporter substrates in TSdb are classified according to their biochemical properties, biological roles and subcellular localizations. In addition to the functional annotation of transporters, extensive compound annotation that includes inhibitor information from the KEGG Ligand and BRENDA databases has been integrated, making TSdb a useful source for the discovery of potential inhibitory mechanisms linking transporter substrates and metabolic enzymes. User-friendly web interfaces are designed for easy access, query and download of the data. Text and BLAST searches against all transporters in the database are provided. We will regularly update the substrate data with evidence from new publications.

A High-performance Liquid Chromatography:chemiluminescence Method for Potential Determination of Vardenafil in Dietary Supplement

A flow method of high-performance liquid chromatography (HPLC) seperation and chemiluminescence (CL) detection for sensitive vardenafil analysis in dietary supplements was developed. The vardenafil separation was achieved on a C18 column at 30°C using ethanol-H(3)PO(4) and ethylenediaminetetraacetic acid disodium salt (Na(2)EDTA) aqueous solution (25 : 75, v/v%) as mobile phase. The followed continuous CL detection was conducted based on the strong CL enhancement by the presence of vardenafil to luminol-K(3)Fe(CN)(6) reaction in alkaline medium. At the flow rate of 0.8 mL/min, the vardenafil retention time (t(R)) was 6.4 min. Factors that affected the HPLC resolution and CL detection were studied and optimized. The calibration curve obtained for vardenafil standard was linear in concentration range of 8.0 × 10(-7) ~ 1.0 × 10(-4) mol/L. The relative standard deviations (RSD) of intraday and interday precision were less than 3.5%. The proposed method was applied to the vardenafil determination in oral liquid, wine, and capsule samples.

Ionic Components of Electric Current at Rat Corneal Wounds

Endogenous electric fields and currents occur naturally at wounds and are a strong signal guiding cell migration into the wound to promote healing. Many cells involved in wound healing respond to small physiological electric fields in vitro. It has long been assumed that wound electric fields are produced by passive ion leakage from damaged tissue. Could these fields be actively maintained and regulated as an active wound response? What are the molecular, ionic and cellular mechanisms underlying the wound electric currents?

Electrically Guiding Migration of Human Induced Pluripotent Stem Cells

A major road-block in stem cell therapy is the poor homing and integration of transplanted stem cells with the targeted host tissue. Human induced pluripotent stem (hiPS) cells are considered an excellent alternative to embryonic stem (ES) cells and we tested the feasibility of using small, physiological electric fields (EFs) to guide hiPS cells to their target. Applied EFs stimulated and guided migration of cultured hiPS cells toward the anode, with a stimulation threshold of <30 mV/mm; in three-dimensional (3D) culture hiPS cells remained stationary, whereas in an applied EF they migrated directionally. This is of significance as the therapeutic use of hiPS cells occurs in a 3D environment. EF exposure did not alter expression of the pluripotency markers SSEA-4 and Oct-4 in hiPS cells. We compared EF-directed migration (galvanotaxis) of hiPS cells and hES cells and found that hiPS cells showed greater sensitivity and directedness than those of hES cells in an EF, while hES cells migrated toward cathode. Rho-kinase (ROCK) inhibition, a method to aid expansion and survival of stem cells, significantly increased the motility, but reduced directionality of iPS cells in an EF by 70-80%. Thus, our study has revealed that physiological EF is an effective guidance cue for the migration of hiPS cells in either 2D or 3D environments and that will occur in a ROCK-dependent manner. Our current finding may lead to techniques for applying EFs in vivo to guide migration of transplanted stem cells.

Paraquat Detoxification with P-sulfonatocalix-[4]arene by a Pharmacokinetic Study

The p-sulfonatocalix[n]arenes are supposed to show potential application in the clinical treatment of viologen poisoning. In the present study, p-sulfonatocalix[4]arene (C4AS), the most common derivative of p-sulfonatocalix[n]arenes, is used to study the antidotic mechanism for paraquat (PQ) by pharmacokinetics in vivo. A high-performance liquid chromatography (HPLC) method was established to determine the concentration of PQ in rat plasma. The results showed that the peak plasma concentration (C(max)) and area under the plasma concentration-time curve (AUC(0-t)) were significantly lower after C4AS intervention than in the PQ intoxication group. It was considered that C4AS has great effective detoxication to PQ poisoning, and the results of in vitro intestinal absorption studies showed that C4AS can inhibit the absorption of PQ via oral administration by forming a stable inclusion constant.

A Pilot Assessment of Relapse Prevention for Heroin Addicts in a Chinese Rehabilitation Center

To conduct a pilot assessment of relapse prevention (RP) group therapy for heroin-dependent patients in a drug rehabilitation center in China.

A Fibrin Gel Loaded with Chitosan Nanoparticles for Local Delivery of RhEGF: Preparation and in Vitro Release Studies

Recombinant human epidermal growth factor (rhEGF) is known to stimulate cell proliferation and accelerate wound healing. Direct delivery of rhEGF at the wound site in a sustained and controllable way without loss of bioactivity would enhance its biological effects. The aim of this study was to prepare a novel local delivery system for the sustained and controllable release of rhEGF, a fibrin gel loaded with chitosan nanoparticles. First, rhEGF-loaded chitosan nanoparticles were prepared and characterized, and these showed an ability to protect rhEGF from proteolysis. The prepared nanoparticles were then incorporated into a fibrin gel matrix during polymerization. In vitro release studies showed that the fibrin gel loaded with rhEGF/chitosan nanoparticles could achieve a more sustained release of rhEGF than either chitosan nanoparticles or an unloaded fibrin gel. Additionally, the release rate could be controlled by altering the contents of fibrinogen and thrombin in this composite delivery system. The bioactivity of the released rhEGF was determined by assessing its ability to stimulate the proliferation of BALB/c 3T3 cells, and the results showed that rhEGF bioactivity was not affected during the preparation process and could be maintained for at least 7 days. This novel delivery system may have great potential applications in the local administration of rhEGF.

Macular Retinoschisis Associated with Normal Tension Glaucoma

To describe a case of macular retinoschisis in a patient with normal tension glaucoma without evidence of optic nerve pits or peripapillary retinoschisis.

A New Approach for Concealed Information Identification Based on ERP Assessment

Recently, numerous concealed information test (CIT) studies have been done with event related potential (ERP) for its sufficient validity in applied use. In this study, a new approach based on wavelet coefficients (WCs) and kernel learning algorithm is proposed to identify concealed information. Totally 16 subjects went through the designed CIT paradigm and the multichannel electroencephalogram (EEG) signals were recorded. Then, the high-dimensional WCs of ERP in delta, theta, alpha and beta rhythms were extracted. For the analysis of the data, kernel principle component analysis (KPCA) and a support vector machines (SVM) classifier are implemented. The results show that WCs features are significant differences between concealed information and irrelevant information (P < 0.05). The KPCA is able to effectively reduce feature dimensionalities and increase generalization performance of SVM. A high accuracy (93.6%) in recognizing concealed information and irrelevant information is achieved, which indicates the combination KPCA and SVM may provide a useful tool for detecting the concealed information.

Calcium Carbonate Phase Transformations During the Carbonation Reaction of Calcium Heavy Alkylbenzene Sulfonate Overbased Nanodetergents Preparation

The preparation and application of overbased nanodetergents with excess alkaline calcium carbonate is a good example of nanotechnology in practice. The phase transformation of calcium carbonate is of extensive concern since CaCO(3) serves both as an important industrial filling material and as the most abundant biomineral in nature. Industrially valuable overbased nanodetergents have been prepared based on calcium salts of heavy alkylbenzene sulfonate by a one-step process under ambient pressure, the carbonation reaction has been monitored by the instantaneous temperature changes and total base number (TBN). A number of analytical techniques such as TGA, DLS, SLS, TEM, FTIR, and XRD have been utilized to explore the carbonation reaction process and phase transformation mechanism of calcium carbonate. An enhanced understanding on the phase transformation of calcium carbonate involved in calcium sulfonate nanodetergents has been achieved and it has been unambiguously demonstrated that amorphous calcium carbonate (ACC) transforms into the vaterite polymorph rather than calcite, which would be of crucial importance for the preparation and quality control of lubricant additives and greases. Our results also show that a certain amount of residual Ca(OH)(2) prevents the phase transformation from ACC to crystalline polymorphs. Moreover, a vaterite nanodetergent has been prepared for the first time with low viscosity, high base number, and uniform particle size, nevertheless a notable improvement on its thermal stability is required for potential applications.

4-Bromo-3-methyl-anilinium Perchlorate 18-crown-6 Clathrate

The reaction of 4-bromo-3-methyl-anilinium perchlorate and 18-crown-6 in methanol solution yielded the title compound, C(7)H(9)BrN(+)·ClO(4) (-)·C(12)H(24)O(6). The protonated 4-bromo-3-methyl-amine unit contains one -NH(3) (+) substituent, resulting in a 1:1 supra-molecular rotator-stator structure, (C(7)H(9)Br-NH(3) (+))(18-crown-6), through three bifurcated N-H⋯(O,O) hydrogen bonds between the ammonium group of the cation and the O atoms of the crown ether mol-ecule.

DC Electric Stimulation Upregulates Angiogenic Factors in Endothelial Cells Through Activation of VEGF Receptors

Small direct current (DC) electric fields direct some important angiogenic responses of vascular endothelial cells. Those responses indicate promising use of electric fields to modulate angiogenesis. We sought to determine the regulation of electric fields on transcription and expression of a serial of import angiogenic factors by endothelial cells themselves. Using semi-quantitative PCR and ELISA we found that electric stimulation upregulates the levels of mRNAs and proteins of a number of angiogenic proteins, most importantly VEGF165, VEGF121 and IL-8 in human endothelial cells. The up-regulation of mRNA levels might be specific, as the mRNA encoding bFGF, TGF-beta and eNOS are not affected by DC electric stimulation at 24h time-point. Inhibition of VEGF receptor (VEGFR1 or VEGFR2) signaling significantly decreased VEGF production and completely abolished IL-8 production. DC electric stimulation selectively regulates production of some growth factors and cytokines important for angiogenesis through a feed-back loop mediated by VEGF receptors.

Endogenous Production of Hydrogen Sulfide in Isolated Bovine Eye

Hydrogen sulfide (H(2)S) is a novel gasotransmitter with physiological and pathological functions in vascular homeostasis, cardiovascular system and central nervous system. In the present study, we determined the endogenous levels of H(2)S in various tissues of the bovine eye. We also examined the basal levels of H(2)S in response to donors (sodium hydrosulfide, NaHS and sodium sulfide, Na(2)S), substrate (L: -cysteine), inhibitors (propargylglycine, PAG and aminooxyacetic acid, AOA) and activator (S-adenosyl-L: -methionine, SAM) of this gas in the bovine retina. H(2)S was measured using a well established spectrophotometric method. The highest concentration of endogenous H(2)S was detected in cornea (19 ± 2.85 nmoles/mg protein, n = 6) and retina (17 ± 2.1 nmoles/mg protein, n = 6). Interestingly, H(2)S was not present in vitreous humor. The inhibitors of CSE and CBS; PAG (1 mM) and AOA (1 mM), significantly attenuated the production of H(2)S in the bovine retina by 56.8 and 42%, respectively. On the other hand the activator of CBS; SAM (100 μM), H(2)S donors; NaHS (1 μM) and Na(2)S (100 μM), significantly increased endogenous levels of H(2)S in bovine retina. L: -cysteine (10-300 μM) produced a significant (P < 0.05) concentration-dependent increase in H(2)S levels reaching a maximal at 300 μM. We conclude that H(2)S is endogenously produced in various tissues of the isolated bovine eye. Moreover, endogenous levels of H(2)S are enhanced in the presence of substrate (L: -cysteine), an activator of CBS (SAM) and H(2)S donors but are blocked by inhibitors of enzymes that synthesize this gas in neural retina.

B and T Lymphocyte Attenuator Down-regulation by HIV-1 Depends on Type I Interferon and Contributes to T-cell Hyperactivation

Nonspecific T-cell hyperactivation is the main driving force for human immunodeficiency virus (HIV)-1 disease progression, but the reasons why the excess immune response is not properly shut off are poorly defined.

[Diagnostic Analysis of Hospitalized Patients with Fever of Unknown Origin at Department of Infectious Diseases]

To explore the etiology, diagnostic methods and procedures for patients with fever of unknown origin (FUO) at department of infectious diseases.

[Amniotic Membrane Transplant Using Fibrin Glue for the Treatment of Deep Layer Corneal Damage]

To treat deep layer corneal damage using fibrin glue and amniotic membrane transplant.

Characterization of PM(2.5) Collected During Broadcast and Slash-pile Prescribed Burns of Predominately Ponderosa Pine Forests in Northern Arizona

Prescribed burning, in combination with mechanical thinning, is a successful method for reducing heavy fuel loads from forest floors and thereby lowering the risk of catastrophic wildfire. However, an undesirable consequence of managed fire is the production of fine particulate matter or PM(2.5) (particles ≤2.5 µm in aerodynamic diameter). Wood-smoke particulate data from 21 prescribed burns are described, including results from broadcast and slash-pile burns. All PM(2.5) samples were collected in situ on day 1 (ignition) or day 2. Samples were analyzed for mass, polycyclic aromatic hydrocarbons (PAHs), inorganic elements, organic carbon (OC), and elemental carbon (EC). Results were characteristic of low intensity, smoldering fires. PM(2.5) concentrations varied from 523 to 8357 µg m(-3) and were higher on day 1. PAH weight percents (19 PAHs) were higher in slash-pile burns (0.21 ± 0.08% OC) than broadcast burns (0.07 ± 0.03% OC). The major elements were K, Cl, S, and Si. OC and EC values averaged 66 ± 7 and 2.8 ± 1.4% PM(2.5), respectively, for all burns studied, in good agreement with literature values for smoldering fires.

MiR-17-92 Cluster MicroRNAs Confers Tumorigenicity in Multiple Myeloma

miRNAs play important roles in the regulation of cell proliferation, differentiation and apoptosis. The deregulation of miRNAs expression contributes to tumorigenesis by modulating oncogenic and tumor suppressor signaling pathways. Oncogenic transcription factor Myc can control expression of a large set of microRNAs (miRNAs). Previous studies have shown that the expression of miR-17-92 cluster, a polycistron encoding six microRNAs (miRNA), has close relationship with the expression of Myc. In current study, silencing Myc in multiple myeloma (MM)cells induced cell death and growth inhibition, and downregulated expression of miR-17-92 cluster. Overexpression of miR-17 or miR-18 could partly abrogated Myc-knockdown-induced MM cell apoptosis. One of the mechanism of Myc inhibiting MM cell apoptosis is through Myc activates miR-17-92 cluster and subsequently down-modulates proapoptotic protein Bim. Although miR-17-92 cluster are located at 13q31.3, the expression of miR-18, miR-19 and miR-20 (especially miR-19) in patients with del(13q14) was higher than those without del(13q14). Patients with miR-17, miR-20 and miR-92 high-expression had shorter PFS compared to those with miR-17, miR-20 and miR-92 low-expression. These results suggest the Myc-inducible miR-17-92 cluster miRNAs contribute to tumorigenesis and poor prognosis in multiple myeloma.

Airway Epithelial Wounds in Rhesus Monkey Generate Ionic Currents That Guide Cell Migration to Promote Healing

Damage to the respiratory epithelium is one of the most critical steps to many life threatening diseases, such as acute respiratory distress syndrome (ARDS) and chronic obstructive pulmonary disease (COPD). The mechanisms underlying repair of the damaged epithelium have not yet been fully elucidated. Here we provide experimental evidence suggesting a novel mechanism for wound repair - endogenous electric currents. It is known that the airway epithelium maintains a voltage difference referred to as the transepithelial potential (TEP). Using a non-invasive vibrating probe, we demonstrate that wounds in the epithelium of trachea from rhesus monkeys generate significant outward electric currents. A small slit wound produced an outward current (1.59 μA/cm(2)), which could be enhanced (nearly doubled) by the ion transport stimulator, aminophylline. In addition, inhibiting cystic fibrosis transmembrane conductance regulator (CFTR) with CFTR(Inh)-172 significantly reduced wound currents (0.17 μA/cm(2)), implicating an important role of ion transporters in wound induced electric potentials. Time-lapse video microscopy showed that applied electric fields (EFs) induced robust directional migration of primary tracheobronchial epithelial cells from rhesus monkeys, towards the cathode, with a threshold of less than 23 mV/mm. Reversal of the field polarity induced cell migration towards the new cathode. We further demonstrate that application of an EF promoted wound healing in a monolayer wound healing assay. Our results suggest that endogenous electric currents at sites of tracheal epithelial injury may direct cell migration, which could benefit restitution of damaged airway mucosa. Manipulation of ion transport may lead to novel therapeutic approaches to repair damaged respiratory epithelium.

Differential Regulations of AQP4 and Kir4.1 by Triamcinolone Acetonide and Dexamethasone in the Healthy and Inflamed Retina

Glucocorticoids are used to treat macular edema, although the mechanisms underlying this effect remain largely unknown. The authors have evaluated in the normal and endotoxin-induced uveitis (EIU) rats, the effects of dexamethasone (dex) and triamcinolone acetonide (TA) on potassium channel Kir4.1 and aquaporin-4 (AQP4), the two main retinal Müller glial (RMG) channels controlling retinal fluid movement.

Different Roles of Membrane Potentials in Electrotaxis and Chemotaxis of Dictyostelium Cells

Many types of cells migrate directionally in direct current (DC) electric fields (EFs), a phenomenon termed galvanotaxis or electrotaxis. The directional sensing mechanisms responsible for this response to EFs, however, remain unknown. Exposing cells to an EF causes changes in plasma membrane potentials (V(m)). Exploiting the ability of Dictyostelium cells to tolerate drastic V(m) changes, we investigated the role of V(m) in electrotaxis and, in parallel, in chemotaxis. We used three independent factors to control V(m): extracellular pH, extracellular [K(+)], and electroporation. Changes in V(m) were monitored with microelectrode recording techniques. Depolarized V(m) was observed under acidic (pH 5.0) and alkaline (pH 9.0) conditions as well as under higher extracellular [K(+)] conditions. Electroporation permeabilized the cell membrane and significantly reduced the V(m), which gradually recovered over 40 min. We then recorded the electrotactic behaviors of Dictyostelium cells with a defined V(m) using these three techniques. The directionality (directedness of electrotaxis) was quantified and compared to that of chemotaxis (chemotactic index). We found that a reduced V(m) significantly impaired electrotaxis without significantly affecting random motility or chemotaxis. We conclude that extracellular pH, [K(+)], and electroporation all significantly affected electrotaxis, which appeared to be mediated by the changes in V(m). The initial directional sensing mechanisms for electrotaxis therefore differ from those of chemotaxis and may be mediated by changes in resting V(m).

Immunologic Changes During Pandemic (H1N1) 2009, China

We analyzed changes in immunologic values over time for 28 hospitalized patients with pandemic (H1N1) 2009. Levels of interleukin-6, interferon-y, and interleukin-10 increased 1 day after illness onset and then decreased to baseline levels. Levels of virus-specific antibody were undetectable 1 day after illness onset and peaked 36 days later.

[Localization of Functional Domains of HEV ORF1 in Cells]

To investigate the expression and localization of various functional domains of ORF1 polyprotein and ORF3 protein of hepatitis E virus in host cells, the coding sequences of the various functional domains (RdRp, HEL, MET, PLP, X) of ORF1 were separately cloned into pcDNA3. 1-GFP vectors for constructing the recombinant plasmids which were verified by enzyme digestion and sequencing. The exact expression of the fusion proteins were detected by Western Blot, and the distribution and localization were observed by the laser scanning confocal microscope(LSCM). In huh7 cells, GFP-RdRp proteins were found mainly in the nuclei, GFP-HEL proteins were distributed vesicularly around the nucleus, GFP-MET proteins were distributed granularly both in the nuclei and the cytoplasm, GFP-PLP proteins had polar distribution around the nucleus, and unknown GFP-X proteins were distributed uniformly both in the nuclei and the cytoplasm. Different localization of these proteins verified the previous data obtained from in vitro studies, providing a support for further research on the biological functions of various proteins coded by HEV genome.

[Expression and Purification of Different Segments from HCoV-NL63 Nucleocapsid Protein and Their Application in Detection of Antibodies]

Prokaryotic expression plasmids carrying N-terminal(1-163aa) and C-terminal(141-306aa) gene of HCoV-NL63 nucleocapsid protein were constructed with pET-30a(+) vector. Consequently, we prepared two purified proteins, Np and Cp, respectively, and established a Western blotting-based line assay (WBLA) for detection of antibodies against HCoV-NL63 using three purified proteins: Np , Cp and Nf, a full-length HCoV-NL63 nucleocapsid protein as previously reported. We detected anti-HCoV-NL63 antibodies among 50 sera samples collected from adult for health-examination by WBLA. The results showed that: 25 (50%), 27 (54%), 36 (72%) of 50 sera were indentified as anti-HCoV-NL63 antibody positive when the antigen was from Nf, Np and Cp, respectively. Among these sera with positive anti-HCoV-NL63 antibody,Cp showed highest antibody positive rate in WBLA,and consistent rates of detection were 64% between Nf and Np, 54% between Nf and Cp, 54% between Np and Cp. Our study provides the foundation for development of HCoV-NL63 serological detection reagents and an experimental tool for immunological research of HCoV-NL63 infection.

[Characterization and Development of Recombinant Vaccinia Viruses Expressing Different Segments of Spike Protein Derived from Human Coronavirus NL-63]

The spike (S) glycoprotein of HCoV-NL63 is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. Four fragments coding spike proteins (S1, S2, RL and RS) from HCoV-NL63 were amplified and cloned into the expression vector derived from vaccinia virus (Tiantan strain), and recombinant vaccinia viruses expressing four segments of spike proteins were generated (vJSC1175-S1; vJSC1175-S2; vJSC1175-RL; vJSC1175-RS), respectively. Their expression location in cell and level were characterized using indirect immune fluorescence assay (IFA) and Western-Blot, respectively. The expressions of four segments of spike proteins in recombinant vaccinia viruses were showed at appropriate level and with posttranslational modification (glycosylation), and S1, RL and RS were mainly distributed in the cell membrane, while the S2 was mainly distributed in the cytoplasm. Our results provide a basis for further exploring diagnostic role and vaccine development of different spike segments from HCoV-NL63.

Effects of a Randomized Contingency Management Intervention on Opiate Abstinence and Retention in Methadone Maintenance Treatment in China

Methadone maintenance treatment has been made available in China in response to the rapid spread of human immunodeficiency virus (HIV), but high rates of dropout and relapse are problematic. The aim of this study was to apply and test if a contingency management (or motivational incentives) intervention can improve treatment retention and reduce drug use.

[Childhood Rhabdomyosarcoma: a Retrospective Review of 23 Cases]

To study the clinical characteristics, treatment and outcome of childhood rhabdomyosarcoma.

Nerve Growth Factor-mediated Neuronal Plasticity in Spinal Cord Contributes to Neonatal Maternal Separation-induced Visceral Hypersensitivity in Rats

Visceral hyperalgesia is a multifactorial gastrointestinal disorder which featured with alterations of abdominal motility and/or gut sensitivity, and is believed to be triggered by environmental stressor or psychological factors. However, its etiology remains incompletely understood. In this study, we aimed to investigate whether nerve growth factor (NGF)-mediated neuronal plasticity is involved in neonatal maternal separation (NMS)-induced visceral hypersensitivity in adult rats, and whether NGF antagonist can attenuate or block such development. In our experiments, animals subjected to NMS were developed with visceral hyperalgesia at age of 8weeks. The threshold for visceral pain among these NMS rats was remarkably lowered than that of the normal handling (NH) rats; however, the expression levels of NGF, c-fos, calcitonin gene-related peptide (CGRP), Substance P, and tyrosine kinases A (TrkA) were notably elevated in lumbosacral spinal cord and/or dorsal root ganglion (DRG) when comparing to those of the NH rats. Further, as intra-peritoneal administration of NGF (10μl at 1μg/kg/day) was given to NH rats during neonatal period, effects that comparable to NMS induction were observed in the adulthood. In contrast, when NMS rats were treated with NGF antagonist K252a (10μl/day from postnatal days 2-14), which acts against tyrosine kinases, the neonatal stress-induced down-shifted visceral pain threshold was restored and neuronal activation, specifically NGF and neuropeptide production, was attenuated. In conclusion, our data strongly suggest that NGF triggers neuronal plasticity and plays a crucial role in NMS-induced visceral hypersensitivity in which NGF antagonism provides positive inhibition via blocking the tyrosine phosphorylation of TrkA.

Role of Novelty Seeking Personality Traits As Mediator of the Association Between COMT and Onset Age of Drug Use in Chinese Heroin Dependent Patients

BACKGROUND: Personality traits such as novelty seeking (NS) are associated with substance dependence but the mechanism underlying this association remains uncertain. Previous studies have focused on the role of the dopamine pathway. OBJECTIVE: Examine the relationships between allelic variants of the catechol-O-methyltransferase (COMT) gene, NS personality traits, and age of onset of drug use in heroin-dependent subjects in China. METHODS: The 478 heroin dependent subjects from four drug rehabilitation centers in Shanghai who were genotyped for eight tagging single nucleotide polymorphisms (SNP) on the COMT gene completed the NS subscale from the Temperament and Character Inventory. Multivariate analyses were used to assess the potential mediating role of NS personality traits in the association between COMT gene variants and the age of onset of heroin use. PRINCIPAL FINDINGS AND CONCLUSIONS: In the univariate analysis the COMT rs737866 gene variants were independently associated with both NS and age of onset of drug use: those with the TT genotype had higher NS subscale scores and an earlier onset age of heroin use than individuals with CT or CC genotypes. In the multivariate analysis the inclusion of the NS subscore variable weakened the relationship between the COMT rs737866 TT genotype and an earlier age of onset of drug use. Our findings that COMT is associated with both NS personality traits and with the age of onset of heroin use helps to clarify the complex relationship between genetic and psychological factors in the development of substance abuse.

Aβ1-16 Can Aggregate and Induce the Production of Reactive Oxygen Species, Nitric Oxide, and Inflammatory Cytokines

Amyloid-β (Aβ40/42) aggregates containing the cross-β-sheet structure are associated with the pathogenesis of Alzheimer's disease (AD). It is generally accepted that the N-terminal peptide of Aβ40/42, Aβ1-16, does not aggregate, and is not cytotoxic. However, we here show that Aβ1-16 can aggregate, and form cytotoxic aggregates containing β-turns and regular non-amyloid β-sheet structures. Factors such as pH, ionic strength, and agitation were found to influence Aβ1-16 aggregation, and the amino acid residues Asp1, His6, Ser8, and Val12 in Aβ1-16 may play a role in this aggregation. Our MTT results showed that Aβ1-16 monomers or oligomers were toxic to SH-SY5Y cells, but Aβ1-16 fibrils exhibited less cytotoxicity. Our studies also indicate that Aβ1-16 aggregates can increase the formation of reactive oxygen species and nitric oxide, induce the loss of calcium homeostasis, and incur the microglial production of TNF-α and IL-4. Thus, our findings suggest that Aβ1-16 may contribute to AD pathogenesis.

[The Early Diagnosis Value of EV 71 IgM Class Antibodies in the Hand, Foot and Mouth Disease]

Assessment of detection of IgM antibodies for human enterovirus 71 (EV 71) in early diagnosis for the hand, foot and mouth disease (HFMD).

Characterization and Dye Decolorization Ability of an Alkaline Resistant and Organic Solvents Tolerant Laccase from Bacillus Licheniformis LS04

A new bacterial strain exhibiting laccase activity was isolated from forest soil and was identified as Bacillus licheniformis LS04. The spore laccase of B. licheniformis LS04 demonstrated a broad pH range for catalyzing substrates. It was quite stable at high temperature and alkaline pH. There was no loss of laccase activity after 10days incubation at pH 9.0, and about 16% of the initial activity was detected after 10h at 80°C. In addition, the spore laccase was tolerant towards 1M of NaCl and 30% of organic solvents. Reactive black 5, reactive blue 19 and indigo carmine were decolorized by the spore laccase in the absence of mediator. Meanwhile, the decolorization process was efficiently promoted when acetosyringone was present, with more than 80% of color removal in 1h at pH 6.6 or 9.0. The unusual properties indicated a high potential in industrial applications for this novel spore laccase.

Globular Adiponectin Protects Human Umbilical Vein Endothelial Cells Against Apoptosis Through Adiponectin Receptor 1/adenosine Monophosphate-activated Protein Kinase Pathway

Endothelial dysfunction is a key event in the onset and progression of atherosclerosis in diabetic patients. Apoptosis may lead to endothelial dysfunction and contribute to vascular complications. However, no study has addressed apoptosis in human umbilical vein endothelial cells (HUVECs) induced by an intermittent high-glucose media and its association with adiponectin receptor 1 (adipoR1), adipoR2, or adenosine monophosphate (AMP)-activated protein kinase (AMPK).

Vesicular Zinc Promotes Presynaptic and Inhibits Postsynaptic Long-term Potentiation of Mossy Fiber-CA3 Synapse

The presence of zinc in glutamatergic synaptic vesicles of excitatory neurons of mammalian cerebral cortex suggests that zinc might regulate plasticity of synapses formed by these neurons. Long-term potentiation (LTP) is a form of synaptic plasticity that may underlie learning and memory. We tested the hypothesis that zinc within vesicles of mossy fibers (mf) contributes to mf-LTP, a classical form of presynaptic LTP. We synthesized an extracellular zinc chelator with selectivity and kinetic properties suitable for study of the large transient of zinc in the synaptic cleft induced by mf stimulation. We found that vesicular zinc is required for presynaptic mf-LTP. Unexpectedly, vesicular zinc also inhibits a form of postsynaptic mf-LTP. Because the mf-CA3 synapse provides a major source of excitatory input to the hippocampus, regulating its efficacy by these dual actions, vesicular zinc is critical to proper function of hippocampal circuitry in health and disease.

[Studies on the Chemical Constituents from the Flowers of Ophiopogon Japonicus]

To study the chemical constituents from the flowers of Ophiopogon japonicus.

Specific Ion Fluxes Generate Cornea Wound Electric Currents

The corneal epithelium generates a significant trans-epithelial potential (TEP) which aids in maintaining cornea water balance and transparency. Injury to the cornea causes a short circuit of the TEP at the wound. The TEP in the intact epithelium around the wound acts like a battery, powering significant ion flux and electric current at the wound. These circulating endogenous currents generate an electric field orientated towards the wound, with the wound the cathode. Many cell types, including human corneal epithelial cells and keratinocytes, migrate to the cathode at physiological electric field strengths. Indeed, the electric signal is a powerful stimulator of cell migration, which appears to override other cues such as chemotaxis and wound void. These wound fields also have a dynamic timecourse of change after wounding. It has been assumed that wound electric fields are produced by passive leakage of ions from damaged cells and tissue. Could these fields be actively maintained and regulated as an active wound response? What are the molecular, ionic and cellular mechanisms underlying the wound electric currents?

[The Studies for Activating and Inhibitory Receptors on Natural Killer Cells in HIV/HCV Co-infected Patients]

To investigate the characteristics of inhibitory and activating receptor expressions on natural killer (NK) cells in HIV/HCV co-infected patients.

[Preparation and Application of a Novel HCV Diagnostic Antigen Fused to Streptavidin]

To prepare streptavidin-tagged hepatitis C virus (HCV) fusion protein and explore its application for the detection of antibody against HCV infection.

Conformation-dependent ScFv Antibodies Specifically Recognize the Oligomers Assembled from Various Amyloids and Show Colocalization of Amyloid Fibrils with Oligomers in Patients with Amyloidoses

Increasing evidence indicates that amyloid aggregates, including oligomers, protofibrils or fibrils, are pivotal toxins in the pathogenesis of many amyloidoses such as Alzheimer's disease (AD), Parkinson's disease, Huntington's disease, prion-related diseases, type 2 diabetes and hereditary renal amyloidosis. Various oligomers assembled from different amyloid proteins share common structures and epitopes. Here we present data indicating that two oligomer-specific single chain variable fragment (scFv) antibodies isolated from a naïve human scFv library could conformation-dependently recognize oligomers assembled from α-synuclein, amylin, insulin, Aβ1-40, prion peptide 106-126 and lysozyme, and fibrils from lysozyme. Further investigation showed that both scFvs inhibited the fibrillization of α-synuclein, amylin, insulin, Aβ1-40 and prion peptide 106-126, and disaggregated their preformed fibrils. However, they both promoted the aggregation of lysozyme. Nevertheless, the two scFv antibodies could attenuate the cytotoxicity of all amyloids tested. Moreover, the scFvs recognized the amyloid oligomers in all types of plaques, Lewy bodies and amylin deposits in the brain tissues of AD and PD patients and the pancreas of type 2 diabetes patients respectively, and showed that most amyloid fibril deposits were colocalized with oligomers in the tissues. Such conformation-dependent scFv antibodies may have potential application in the investigation of aggregate structures, the mechanisms of aggregation and cytotoxicity of various amyloids, and in the development of diagnostic and therapeutic reagents for many amyloidoses.

MicroRNA Expression and Regulation in Human, Chimpanzee, and Macaque Brains

Among other factors, changes in gene expression on the human evolutionary lineage have been suggested to play an important role in the establishment of human-specific phenotypes. However, the molecular mechanisms underlying these expression changes are largely unknown. Here, we have explored the role of microRNA (miRNA) in the regulation of gene expression divergence among adult humans, chimpanzees, and rhesus macaques, in two brain regions: prefrontal cortex and cerebellum. Using a combination of high-throughput sequencing, miRNA microarrays, and Q-PCR, we have shown that up to 11% of the 325 expressed miRNA diverged significantly between humans and chimpanzees and up to 31% between humans and macaques. Measuring mRNA and protein expression in human and chimpanzee brains, we found a significant inverse relationship between the miRNA and the target genes expression divergence, explaining 2%-4% of mRNA and 4%-6% of protein expression differences. Notably, miRNA showing human-specific expression localize in neurons and target genes that are involved in neural functions. Enrichment in neural functions, as well as miRNA-driven regulation on the human evolutionary lineage, was further confirmed by experimental validation of predicted miRNA targets in two neuroblastoma cell lines. Finally, we identified a signature of positive selection in the upstream region of one of the five miRNA with human-specific expression, miR-34c-5p. This suggests that miR-34c-5p expression change took place after the split of the human and the Neanderthal lineages and had adaptive significance. Taken together these results indicate that changes in miRNA expression might have contributed to evolution of human cognitive functions.

Ion-selective Self-referencing Probes for Measuring Specific Ion Flux

The metal vibrating probe developed in the 1970s to measure electric current is sensitive down to the micro-Amp range, but detects only net current due to flow of multiple ions and is too large to measure from single cells. Electrophysiological techniques which use glass microelectrodes such as voltage clamping can be used on single cells but are also non-specific. Ion-selective probes are glass microelectrodes containing at their tip a small amount of ionophore permeable to a particular ion. The electrode is therefore sensitive to changes in concentration of this ion. If the probe tip is moved at low frequency between two points in a concentration gradient of this ion then the electrochemical potential of the solution inside the electrode fluctuates in proportion to the size of the ion gradient. This fluctuation is amplified and recorded and is used to calculate the actual ion flux using Fick's law of diffusion. In this mini-review we describe the technique of ion-selective self-referencing microelectrodes to measure specific ion fluxes. We discuss the development of the technique and describe in detail the methodology and present some representative results.

Expression of Pro- and Anti-angiogenic Isoforms of VEGF in the Mouse Model of Oxygen-induced Retinopathy

Retinopathy of prematurity (ROP) has become one of the leading causes of blindness and visual loss in children over the last half century. Vascular endothelial growth factor (VEGF-A) is the principal stimulator of angiogenesis. Recently, it has been identified that VEGF was differentially spliced from exons 8 to exons 8a and 8b to form two families: the pro-angiogenic VEGFxxx family and the anti-angiogenic VEGFxxxb family. This alternate splicing produced VEGFxxxb proteins of the same length as VEGFxxx family, but with different C terminal amino acid sequences. VEGFxxxb appeared to be able to inhibit VEGFxxx-dependent angiogenesis. In our study, we investigated the protein expression course of VEGFxxx and VEGFxxxb by Western-blot in a mouse model of Oxygen-induced Retinopathy (OIR) from postnatal day 1 (P1) to postnatal day 21 (P21). We also analyzed the relative protein expression level of VEGF(165)b isoform in the OIR mouse model. We found that both VEGFxxx and VEGFxxxb were present in the mouse retina, among which, VEGF(164) and VEGF(165)b appeared to be predominant VEGFxxx and VEGFxxxb isoforms respectively in the mouse retina. We also found that the two family had different expression pattern correlated with neovascularization development and that the relative expression level of VEGF(165)b isoform switched during the neovascularization development in the OIR mouse model. In OIR group, the protein level of total VEGF isoforms (a mix of VEGF(164) and VEGF(165)b, detected by pan-VEGF antibody) continuously increased and peaked at P17 while VEGF(165)b continuously decreased from P9 which was well related with the vessel obliteration and neovascularization development in the mouse model of OIR. The neovascularization development correlates with an increase of total VEGF isoforms and the decrease of VEGF(165)b, indicating that there is a pro-angiogenic VEGF shift. Therefore, anti-angiogenic therapy that could alter the ratio of VEGFxxxb/VEGFxxx may be more effective.

[Study on Alternative Methods for the Acute Oral Toxicity in Detection of Chemicals]

To evaluate three alternative methods for LD50 test-Fixed Dose Procedure (FDP), the Acute Toxic Class Method (ATC) and Up and Down Procedure (UDP).

[Alterations of NK Cell Frequency and Function in HIV/HCV Co-infected Patients]

To study the impacts of HIV/HCV co-infection to NK cells by investigating the changes of frequencies and functions of NK cells in HIV/HCV co-infected patients.

[Clinicopathologic Correlation Between CD4-positive T Lymphocyte Counts and Superficial Lymphadenopathy in HIV-positive/AIDS Patients]

To explore the clinicopathological correlation between CD4(+) T lymphocyte count and superficial lymphadenopathy HIV/AIDS patients.

Cue-induced Craving and Physiological Reactions in Recently and Long-abstinent Heroin-dependent Patients

OBJECTIVE: To understand the different patterns of cue-induced craving and physiological reactions among recently abstinent and long-abstinent heroin-dependent patients. METHOD: 26 healthy adult controls (HC), 29 long-abstinent (more than 1year, LA), and 26 recently abstinent (less than 1month, RA) heroin-dependent individuals were exposed to heroin-related and neutral video cues, one video per session, on different days in random order. Self-reported heroin craving by a 10-point visual analog scale (VAS), physiological reactions [skin conductance (SC), muscle electromyography (MEG), skin temperature (TEMP)] and cardiovascular arousal [heart rates (HR), systolic blood pressure (HBP) and diastolic blood pressure (LBP)] were assessed at baseline and after exposure. RESULTS: Both heroin-abstinent groups showed increased heroin craving, SC, MEG, HR, SBP and LBP after exposure to heroin-related video, compared to the control group and compared to exposure to the neutral video. Except the RA group showed more HR changes, changes of heroin craving, SC, MEG, HR, SBP and LBP after exposure to the heroin cue video were not different between the LA and RA groups. CONCLUSIONS: Abstinent heroin-dependent patients had elevated craving and physiological reactions after exposure to videos containing heroin-related cues and the cue induced responses still occurred in long-abstinent patients. This phenomenon should be addressed in treatment and recovery services for heroin dependence.

Evaluation of the Third-party Mediation Mechanism for Medical Disputes in China

Medical disputes have been increasing in recent years in China, which cause growing tension between doctors and patients. In many locations, it has started as a practice of exploring diversified dispute settlement methods. Great importance has been attached to the non-lawsuit model through third-party mediation, which might have been led by professional organizations, insurance companies, People's Mediation Committees, or three-level governmental authorities. Those have contributed to a rapid effective resolution of medical disputes. However, there are some deficiencies that need to be addressed and fixed up, thus calling for improvement, such as the lack of a sustainable supporting mechanism, unclear legal status of the mediation institutions and mediation agreements, patching up a quarrel by only compensation.

Protective Effect of Paeoniflorin Against Oxidative Stress in Human Retinal Pigment Epithelium in Vitro

This study was conducted to determine whether paeoniflorin (PF) could prevent Hâ‚‚Oâ‚‚-induced oxidative stress in ARPE-19 cells and to elucidate the molecular pathways involved in this protection.

[Clinical Characteristics and Changes in Blood Electrolyte and Renal Function of Neonates with Polycythemia at High Altitude]

To analyze the relationship between neonatal hypoxia and polycythemia and to study clinical characteristics of Tibetan neonates whose family lived in Tibetan plateau for generations and Han neonates whose family moved to the plateau.

Electrical Signaling in Control of Ocular Cell Behaviors

Epithelia of the cornea, lens and retina contain a vast array of ion channels and pumps. Together they produce a polarized flow of ions in and out of cells, as well as across the epithelia. These naturally occurring ion fluxes are essential to the hydration and metabolism of the ocular tissues, especially for the avascular cornea and lens. The directional transport of ions generates electric fields and currents in those tissues. Applied electric fields affect migration, division and proliferation of ocular cells which are important in homeostasis and healing of the ocular tissues. Abnormalities in any of those aspects may underlie many ocular diseases, for example chronic corneal ulcers, posterior capsule opacity after cataract surgery, and retinopathies. Electric field-inducing cellular responses, termed electrical signaling here, therefore may be an unexpected yet powerful mechanism in regulating ocular cell behavior. Both endogenous electric fields and applied electric fields could be exploited to regulate ocular cells. We aim to briefly describe the physiology of the naturally occurring electrical activities in the corneal, lens, and retinal epithelia, to provide experimental evidence of the effects of electric fields on ocular cell behaviors, and to suggest possible clinical implications.

Conversion of Non-endocrine Human Pancreatic Cells to Insulin-producing Cells for Treatment of Diabetes

Type I diabetes results from the autoimmune destruction of the insulin-secreting pancreatic β-cells, affecting many millions of people worldwide. The optimal treatment is to restore the endogenous supply of insulin either through the transplantation of pancreas or the transplantation of islets of langerhans or simply the β-cells. However, the donated pancreas organs are limited and the available organs are only able to treat a small portion of the diabetes patients. Thus, glucose-responsive, insulin-producing cells from human origin are urgently needed. The aim of this chapter is to give some insight views to how to turn the potential human pancreatic non-endocrine cells into cells that are capable of secreting insulin in response to glucose and ameliorating insulin-deficient diabetes conditions after transplantation.

Guided Migration of Neural Stem Cells Derived from Human Embryonic Stem Cells by an Electric Field

Small direct current (DC) electric fields (EFs) guide neurite growth and migration of rodent neural stem cells (NSCs). However, this could be species dependent. Therefore, it is critical to investigate how human NSCs (hNSCs) respond to EF before any possible clinical attempt. Aiming to characterize the EF-stimulated and guided migration of hNSCs, we derived hNSCs from a well-established human embryonic stem cell line H9. Small applied DC EFs, as low as 16 mV/mm, induced significant directional migration toward the cathode. Reversal of the field polarity reversed migration of hNSCs. The galvanotactic/electrotactic response was both time and voltage dependent. The migration directedness and distance to the cathode increased with the increase of field strength. (Rho-kinase) inhibitor Y27632 is used to enhance viability of stem cells and has previously been reported to inhibit EF-guided directional migration in induced pluripotent stem cells and neurons. However, its presence did not significantly affect the directionality of hNSC migration in an EF. Cytokine receptor [C-X-C chemokine receptor type 4 (CXCR4)] is important for chemotaxis of NSCs in the brain. The blockage of CXCR4 did not affect the electrotaxis of hNSCs. We conclude that hNSCs respond to a small EF by directional migration. Applied EFs could potentially be further exploited to guide hNSCs to injured sites in the central nervous system to improve the outcome of various diseases. STEM CELLS 2012; 30:349-355.

Directing Migration of Endothelial Progenitor Cells with Applied DC Electric Fields

Naturally-occurring, endogenous electric fields (EFs) have been detected at skin wounds, damaged tissue sites and vasculature. Applied EFs guide migration of many types of cells, including endothelial cells to migrate directionally. Homing of endothelial progenitor cells (EPCs) to an injury site is important for repair of vasculature and also for angiogenesis. However, it has not been reported whether EPCs respond to applied EFs. Aiming to explore the possibility to use electric stimulation to regulate the progenitor cells and angiogenesis, we tested the effects of direct-current (DC) EFs on EPCs. We first used immunofluorescence to confirm the expression of endothelial progenitor markers in three lines of EPCs. We then cultured the progenitor cells in EFs. Using time-lapse video microscopy, we demonstrated that an applied DC EF directs migration of the EPCs toward the cathode. The progenitor cells also align and elongate in an EF. Inhibition of vascular endothelial growth factor (VEGF) receptor signaling completely abolished the EF-induced directional migration of the progenitor cells. We conclude that EFs are an effective signal that guides EPC migration through VEGF receptor signaling in vitro. Applied EFs may be used to control behaviors of EPCs in tissue engineering, in homing of EPCs to wounds and to an injury site in the vasculature.

AutismKB: an Evidence-based Knowledgebase of Autism Genetics

Autism spectrum disorder (ASD) is a heterogeneous neurodevelopmental disorder with a prevalence of 0.9-2.6%. Twin studies showed a heritability of 38-90%, indicating strong genetic contributions. Yet it is unclear how many genes have been associated with ASD and how strong the evidence is. A comprehensive review and analysis of literature and data may bring a clearer big picture of autism genetics. We show that as many as 2193 genes, 2806 SNPs/VNTRs, 4544 copy number variations (CNVs) and 158 linkage regions have been associated with ASD by GWAS, genome-wide CNV studies, linkage analyses, low-scale genetic association studies, expression profiling and other low-scale experimental studies. To evaluate the evidence, we collected metadata about each study including clinical and demographic features, experimental design and statistical significance, and used a scoring and ranking approach to select a core data set of 434 high-confidence genes. The genes mapped to pathways including neuroactive ligand-receptor interaction, synapse transmission and axon guidance. To better understand the genes we parsed over 30 databases to retrieve extensive data about expression patterns, protein interactions, animal models and pharmacogenetics. We constructed a MySQL-based online database and share it with the broader autism research community at http://autismkb.cbi.pku.edu.cn, supporting sophisticated browsing and searching functionalities.

Microarray-based Mutation Detection of Pediatric Sporadic Nonsyndromic Hearing Loss in China

To investigate the molecular etiologic causes of sporadic nonsyndromic hearing loss in Chinese children.

Hepatitis C Knowledge and Alcohol Consumption Among Patients Receiving Methadone Maintenance Treatment in Shanghai, China

Objective: The aim was to investigate hepatitis C virus (HCV) knowledge and alcohol consumption among patients (n = 114) in a methadone maintenance treatment (MMT) clinic in Shanghai. Methods: A cross-sectional survey was carried out in an MMT clinic. Structured questionnaires (HCV Knowledge Scale and Alcohol Use Disorders Identification Test (AUDIT)) and some open-ended questions were used to assess (i) HCV knowledge, (ii) HCV treatment received, (iii) awareness of HCV status, and (iv) alcohol consumption. Results: Findings revealed the HCV-positive rate was 57.0%. There were significant gaps in knowledge about HCV and HCV treatment received. The group mean score of HCV knowledge was 11.3 out of 20 (SD = 2.1) and the mean score on the AUDIT was 3.2 (SD = 5.4). Most participants (68.4%) reported not knowing their HCV status. Among HCV-positive participants, only 15.3% had received HCV antivirus treatment and 18.4% expressed a need for counseling about HCV infection. Conclusions: Considering the limited HCV knowledge and low level of HCV treatment received, effective HCV education and intervention strategies should be developed to target patients in China's MMT clinics. Moreover, alcohol screening should also be part of the routine assessments within MMT programs. Scientific Significance: This study reveals the importance of HCV testing and education among drug users in MMT clinics.

Electroencephalogram and Electrocardiograph Assessment of Mental Fatigue in a Driving Simulator

Mental fatigue is a contributing factor to some serious transportation crashes. In this study, we measured mental fatigue in drivers using electroencephalogram (EEG) and electrocardiograph (ECG). Together, thirteen healthy subjects performed a continuous simulated driving task for 90min with simultaneous ECG and multi-channel EEG recording of each subject. Several important physiological parameters were investigated using preprocessed ECG and EEG signals. The results show that the EEG alpha and beta, the relative power, the amplitude of P300 wave of event-related potential (ERP), the approximated entropy of the ECG, and the lower and upper bands of power of heart rate variability (HRV) are significantly different before and after finishing the driving task (p<0.05). These metrics are possible indices for measuring simulated driving mental fatigue.

Interaction Between Hedgehog Signalling and PAX6 Dosage Mediates Maintenance and Regeneration of the Corneal Epithelium

To investigate the roles of intracellular signaling elicited by Hedgehog (Hh) ligands in corneal maintenance and wound healing.

Intrinsic Peroxidase-like Activity and Catalase-like Activity of Co(3)O(4) Nanoparticles

We demonstrate that Co(3)O(4) nanoparticles (NPs) exhibit intrinsic peroxidase-like activity and catalase-like activity. The peroxidase-like activity of the Co(3)O(4) NPs originates from their ability of electron transfer between reducing substrates and H(2)O(2), not from ˙OH radical generated. As peroxidase mimetics, Co(3)O(4) NPs were used for colorimetric determination of H(2)O(2) and glucose.

Diurnal and Nocturnal Variations in Aqueous Humor Dynamics of Patients With Ocular Hypertension Undergoing Medical Therapy

OBJECTIVE: To evaluate the interaction of intraocular pressure (IOP)-lowering medications with physiologic day and night changes in aqueous humor dynamics in participants with ocular hypertension. METHODS: Thirty participants were enrolled in this double-masked, randomized, crossover study. Each participant underwent aqueous humor dynamics measurements at baseline and at 2 weeks of dosing in random order with latanoprost in the evening and placebo in the morning, timolol maleate twice daily, and dorzolamide hydrochloride twice daily. Measurements included central corneal thickness by ultrasound pachymetry, anterior chamber depth by A-scan, seated and habitual IOP by pneumatonometry, blood pressure by sphygmomanometry, episcleral venous pressure by venomanometry, and aqueous flow by fluorophotometry. Outflow facility was assessed by fluorophotometry and by tonography. Uveoscleral outflow was mathematically calculated using the Goldmann equation. RESULTS: Latanoprost use significantly decreased IOP during the day and night. It increased daytime uveoscleral outflow by a mean (SD) of 0.90 (1.46) μL/min (P = .048), but a nighttime increase of 0.26 (1.10) μL/min (P = .47) did not reach statistical significance. Timolol use decreased IOP during the day by reducing aqueous flow by 25%. Dorzolamide use lowered IOP only at the noon measurement and reduced daytime aqueous flow by 16%. Neither dorzolamide nor timolol use added to the physiologic 47% reduction in nighttime aqueous flow. CONCLUSIONS: The daytime IOP-lowering effects of latanoprost are mediated by an increase in uveoscleral outflow, and those of timolol and dorzolamide are mediated by aqueous flow suppression. Nighttime physiologic changes in uveoscleral outflow limit the nighttime pharmacodynamic efficacy of latanoprost. Aqueous flow suppression with timolol and dorzolamide was ineffective in obtaining IOP lowering at night. Trial Registration  clinicaltrials.gov Identifier: NCT00572936.

[The Comparison Between HIV-infected Patients' Vdelta2 T Cells Expansion Efficiencies by Zoledronic Acid and Gammadelta TCR Monoclonal Antibody in Vitro]

To find out the more efficient induction method through investigating the expansion efficiencies of HIV-infected patients' Vdelta2 T cells induced by zoledronic acid (Zol) or gammadelta TCR monoclonal antibody (mAb).

Cytotoxic and Antibacterial Cembranoids from a South China Sea Soft Coral, Lobophytum Sp

Chemical examination of a South China Sea soft coral Lobophytum sp. led to the isolation of three new α-methylene-γ-lactone-containing cembranoids, (1R*,3R*, 4R*,14R*,7E,11E)-3,4-epoxycembra-7,11,15(17)-trien-16,14-olide (1), (1R*,7S*,14S*,3E, 11E)-7-hydroperoxycembra-3,8(19),11,15(17)-tetraen-16,14-olide (2), and (1R*,7S*,14S*, 3E,11E)-18-acetoxy-7-hydroperoxycembra-3,8(19),11,15(17)-tetraen-16,14-olide (3), along with eleven known analogues 4-14. The structures of the new compounds were elucidated through extensive spectroscopic analysis, including 1D and 2D NMR data. Compounds 1-3 exhibited moderate cytotoxic activity against the selected tumor cell lines. Moreover, 2 and 3 were found to be moderate inhibitors against the bacteria S. aureus and S. pneumoniae.

P42.3: a Promising Biomarker for the Progression and Prognosis of Human Colorectal Cancer

As a novel cell cycle-related gene, p42.3 has been shown to play a key role in the cell proliferation and tumorigenicity of gastric cancer. To date, the association between p42.3 and colorectal cancer (CRC) has not been reported. This study investigated the expression of p42.3 and its potential role in human colorectal cancers.

Synchronization Modulation Increases Transepithelial Potentials in MDCK Monolayers Through Na/K Pumps

Transepithelial potential (TEP) is the voltage across a polarized epithelium. In epithelia that have active transport functions, the force for transmembrane flux of an ion is dictated by the electrochemical gradient in which TEP plays an essential role. In epithelial injury, disruption of the epithelial barrier collapses the TEP at the wound edge, resulting in the establishment of an endogenous wound electric field (∼100 mV/mm) that is directed towards the center of the wound. This endogenous electric field is implicated to enhance wound healing by guiding cell migration. We thus seek techniques to enhance the TEP, which may increase the wound electric fields and enhance wound healing. We report a novel technique, termed synchronization modulation (SM) using a train of electric pulses to synchronize the Na/K pump activity, and then modulating the pumping cycles to increase the efficiency of the Na/K pumps. Kidney epithelial monolayers (MDCK cells) maintain a stable TEP and transepithelial resistance (TER). SM significantly increased TEP over four fold. Either ouabain or digoxin, which block Na/K pump, abolished SM-induced TEP increases. In addition to the pump activity, basolateral distribution of Na/K pumps is essential for an increase in TEP. Our study for the first time developed an electrical approach to significantly increase the TEP. This technique targeting the Na/K pump may be used to modulate TEP, and may have implication in wound healing and in diseases where TEP needs to be modulated.

3D Arrays for High Throughput Assay of Cell Migration and Electrotaxis

Cell behaviour in 3D environments can be significantly different from those in 2D cultures. With many different 3D matrices being developed and many experimental modalities used to modulate cell behaviour in 3D, it is necessary to develop high throughput techniques to study behaviour in 3D. We report on a 3D array on slide and have adapted this to our electrotaxis chamber, thereby offering a novel approach to quantify cellular responses to electric fields (EFs) in 3D conditions, in different matrices, with different strains of cells, under various field strengths. These developments used Dictyostelium cells to illustrate possible applications and limitations.

Inhibition of CK2 Enhances UV-triggered Apoptotic Cell Death in Lung Cancer Cell Lines

Lung cancer is a high-grade malignancy with poor 5 year-survival rates that remains incurable with current therapies. Different cellular stresses, including antitumor agents, ionizing radiation and ultraviolet (UV) light, can induce apoptosis and activate signaling pathways. UV has multiple effects on tumor cells, including DNA damage, and increases the expression of some genes involved in tumor cell apoptosis and DNA repair. It has been reported that UV can also activate casein kinase 2 (CK2). CK2, a Ser/Thr protein kinase, has been reported to be frequently overexpressed in various types of human cancer, including lung cancer, and is associated with tumor development. Thus, combination of UV and CK2 inhibitors may be a new strategy for the treatment of lung cancer. Our results demonstrated that inhibition of CK2a through CK2 siRNA or a CK2 inhibitor [(4,5,6,7-tetrabromobenzotriazole (TBB)] enhances the decrease in cell viability of lung cancer cells (A549 and H2030) induced by UV. Western blot analysis demonstrated that the combination increased the expression of apoptotic protein markers cytochrome c and the cleavage of poly ADP-ribose polymerase (PARP) and caspase-3. Furthermore, our results indicated that UV decreased the expression of the tumor suppressor protein PML through activation of CK2. Inhibition of CK2 by CK2 siRNA and TBB can recover the reduction of PML induced by UV. Collectively, these results demonstrate the significant apoptosis of lung cancer cells induced by combination treatment of the CK2 inhibitor and UV radiation. CK2 enhanced cell apoptosis by UV radiation may due, at least partly, to recover the expression of PML. These findings warrant the clinical testing of CK2 inhibitors which, when used in conjunction with DNA-damaging agents such as radiation, may be an effective cancer therapeutic strategy.

Structure of RSV Fusion Glycoprotein Trimer Bound to a Prefusion-specific Neutralizing Antibody

The prefusion state of respiratory syncytial virus (RSV) fusion (F) glycoprotein is the target of most RSV-neutralizing activity in human sera, but its metastability has hindered characterization. To overcome this obstacle, we identified prefusion-specific antibodies that were substantially more potent than the prophylactic antibody palivizumab. The cocrystal structure for one of these antibodies, D25, in complex with the F glycoprotein revealed D25 to lock F in its prefusion state by binding to a quaternary epitope at the trimer apex. Electron microscopy showed that two other antibodies, AM22 and 5C4, also bound to the newly identified site of vulnerability, which we named antigenic site Ø. These studies should enable design of improved vaccine antigens and define new targets for passive prevention of RSV-induced disease.

[Immunophenotype Analysis of Leukemic Mantle Cell Lymphoma]

Mantle cell lymphoma (MCL) is a kind of mature B-cell neoplasms with significantly poor prognosis and is usually misdiagnosed. With the development of flow cytometry and cytogenetic technique, most patients were at leukemic phase when diagnosed. This study was purposed to investigate the immunophenotypes of MCL, the immunophenotype information of 22 leukemic MCL patients was analyzed retrospectively. All the patients were conformed t(11;14) translocation by fluorescence in situ hybridization. Immunophenotypes were detected by a four-color flow cytometry including CD3, CD4, CD5, CD8, CD10, CD19, CD20, CD22, CD23, CD25, CD38, CD103, CD148, CD200, FMC7, ZAP-70, κ, λ. The results showed that CD19, CD5, CD20 and monoclonal sIg expressed in all 22 patients with CD20 high expression; CD22 expressed weakly in 17 patients; CD23 expressed in 6 patients including 2 cases highly expressed; FMC7 expressed in 12 patients. 5 patients were 4-point score and 17 patients had a score less than 4 according to CLL scoring system. CD148 and CD200 were detected in 18 patients, in which CD200 expressed negatively in 11 patients, CD200 expressed weakly in 7 patients with median fluorescence intensity (MFI) 25.8 (6.6 - 254.26); CD148 expressed positively in all 18 patients with median MFI: 337 (73.4 - 1341.9). It is concluded that the atypical immunophenotype is common in leukemia MCL, thereby the diagnosis of MCL needs comprehensively analyze with morphocytology, immunophenotype and cytogenetic, CD200 and CD148 as new bio-markers can differentiate MCL from chronic B cell lymphoproliferative disease.

Supersandwich-type Electrochemiluminescenct Aptasensor Based on Ru(phen)3(2+) Functionalized Hollow Gold Nanoparticles As Signal-amplifying Tags

An electrochemiluminescent (ECL) aptasensor was fabricated and used for the amplified detection of thrombin (TB) based on DNA supersandwich structure. Herein, hollow gold nanospheres (HGNPs) were firstly employed as effective tag-carriers for the immobilization of detection aptamer (TBA 2) to form the HGNPs labeled TBA 2 (HGNPs-TBA 2). Subsequently, streptavidin (SA) was used to block the non-specific binding sites of HGNPs-TBA 2 as well as to supply binding sites, which could further introduce numerous initiator DNA strands (bio-S1) via biotin-streptavidin specific interaction. Next, bio-S1 could in situ trigger hybridization chain reaction (HCR) to create a long nicked double helices analogous (dsDNA) in the present of ssDNA S2 and ssDNA S3 (S3 is partially complementary to the S2) to obtain the DNA supersandwich structure. Furthermore, Ru(phen)3(2+), a well-known ECL luminophore, could be intercalated into the grooves of dsDNA (Ru-dsDNA) to form the Ru-dsDNA-SA-HGNPs-TBA 2 bioconjugate. As a result, the target of TB was sandwiched between Ru-dsDNA-SA-HGNPs-TBA 2 and TBA 1. In this strategy, numerous Ru(phen)3(2+) could be immobilized on the electrode based on the supersandwich structure, resulting in an increased ECL signal output. A supersandwich ECL assay for TB detection was developed with excellent sensitivity of a large concentration variation from 5fM to 50pM and a detection limit of 1.6fM.

The Dopamine Receptor D1 Gene is Associated with the Length of Interval Between First Heroin Use and Onset of Dependence in Chinese Han Heroin Addicts

Previous researches showed that the dopamine receptor D1 (DRD1) may play a critical role in drug dependence. This research aimed to determine whether DRD1 played a role in development of heroin dependence in Chinese heroin-dependent patients. 465 Chinese Han heroin-dependent subjects and 379 healthy controls were recruited in the Shanghai region. Five single-nucleotide-polymorphisms (SNPs) of the DRD1 gene were genotyped in all subjects. The results found that the frequencies of DRD1 SNP genotypes or haplotypes were not different between heroin-dependent patients and controls. Among heroin-dependent patients, subjects with rs5326CC and/or rs6882300AA genotypes develop to heroin-dependent more rapidly than those without rs5326CC and/or rs6882300AA genotypes. The results indicated that DRD1 gene polymorphism may not play an important role in the susceptibility of heroin dependence in the Chinese Han population, but it may be associated with the rapidity of heroin dependence development from first drug use.

Role of EuroSCORE II in Predicting Long-term Outcome After Percutaneous Catheter Intervention for Coronary Triple Vessel Disease or Left Main Stenosis

The SYNTAX score (Ssc) assessing the complexity of coronary anatomy predicts survival after percutaneous coronary intervention (PCI). We tested the hypothesis that the newly developed euroSCORE II (eSC2) can improve the prediction of outcome after complex PCI by the Ssc.

Ellagic Acid Inhibits Human Pancreatic Cancer Growth in Balb C Nude Mice

Ellagic acid (EA) is a polyphenol found in several plants and fruits. The objectives of this study were to examine the molecular mechanisms by which EA inhibits pancreatic cancer growth in Balb C nude mice. PANC-1 cells were injected subcutaneously into Balb c nude mice, and tumor-bearing mice were treated with EA. The expression of Akt, Shh and Notch and their target gene products were measured by the immunohistochemistry and Western blot analysis. Treatment of PANC-1 xenografted mice with EA resulted in significant inhibition in tumor growth which was associated with suppression of cell proliferation and caspase-3 activation, and induction of PARP cleavage. EA inhibited the expression of Bcl-2, cyclin D1, CDK2, and CDK6, and induced the expression of Bax in tumor tissues compared to untreated control group. EA inhibited the markers of angiogenesis (COX-2, HIF1α, VEGF, VEGFR, IL-6 and IL-8), and metastasis (MMP-2 and MMP-9) in tumor tissues. Furthermore, treatment of mice with EA caused a significant inhibition in phospho-Akt, Gli1, Gli2, Notch1, Notch3, and Hey1. EA also reversed epithelial to mesenchymal transition by up-regulating E-cadherin and inhibiting the expression of Snail, MMP-2 and MMP-9. These data suggest that EA can inhibit pancreatic cancer growth, angiogenesis and metastasis by suppressing Akt, Shh and Notch pathways. In view of the fact that EA could effectively inhibit human pancreatic cancer growth by suppressing Akt, Shh and Notch pathways, our findings suggest that the use of EA would be beneficial for the management of pancreatic cancer.

SKP2 High Expression, KIT Exon 11 Deletions, and Gastrointestinal Bleeding As Predictors of Poor Prognosis in Primary Gastrointestinal Stromal Tumors

Considering the indication of adjuvant therapy, the recurrence risk for primary gastrointestinal stromal tumor (GIST) after surgery needs to be accurately estimated. However, current risk stratification schemes may still have room for improvement. This study seeks to analyze prognostic factors for primary GISTs from 3 aspects, including clinicopathological parameters, immunohistochemical biomarkers, and gene mutational status, and attempts to find novel valuable factors predicting the malignancy potential of GISTs.

[Environmental Impacts of Sewage Treatment System Based on Emergy Analysis]

"Integrated sewage treatment system" (ISTS) consists of sewage treatment plant system and their products (treated water and dewatered sludge) disposal facilities, which gives a holistic view of the whole sewage treatment process. During its construction and operation, ISTS has two main impacts on the environment, i.e., the consumption of resources and the damage of discharged pollutants on the environment, while the latter was usually ignored by the previous researchers when they assessed the impacts of wastewater treatment system. In order to more comprehensively understanding the impacts of sewage treatment on the environment, an analysis was made on the ISTS based on the theories of emergy analysis, and, in combining with ecological footprint theory, the sustainability of the ISTS was also analyzed. The results showed that the emergy of the impacts of water pollutants on the environment was far larger than that of the impacts of air pollutants, and NH3-N was the main responsible cause. The emergy consumption of ISTS mainly came from the emergy of wastewater and of local renewable resources. The "sewage treatment plant system + landfill system" had the highest emergy utilization efficiency, while the "sewage treatment plant system + reclaimed water reuse system + incineration system" had the lowest one. From the aspect of environmental sustainability, the "sewage treatment plant system + reclaimed water reuse system + landfill system" was the best ISTS, while the "sewage treatment plant system + incineration system" was the worst one.

Acanthamoeba Migration in an Electric Field

We investigated the in vitro response of Acanthamoeba trophozoites to electric fields (EFs).

Mineralocorticoid Receptor Antagonism in the Treatment of Chronic Central Serous Chorioretinopathy: a Pilot Study

Based on experimental data showing that central serous chorioretinopathy could result from overactivation of mineralocorticoid receptor pathway in choroid vessels, the authors studied eplerenone, a mineralocorticoid receptor antagonist, as a potential treatment for chronic central serous chorioretinopathy.

Laboratory Features Throughout the Disease Course of Influenza A (H1N1) Virus Infection

Influenza has emerged every year but a complete profile of laboratory indices throughout the disease course remains unknown.

Anti-angiogenic Effect of KH902 on Retinal Neovascularization

KH902 is a fusion protein derived from the extracellular domains of vascular endothelial growth factor (VEGF) receptors 1 and 2 and the Fc portion of immunoglobulin G1 (IgG1). Retinopathy of prematurity (ROP) is an eye disease that affects premature babies who have received intensive neonatal care, and the disorganization of retinal blood vessels may result in scarring and retinal detachment. This study was designed to examine the inhibitory effects of KH902 on mice with oxygen-induced retinopathy (OIR), one of the animal models of ROP.

Combined Use of Ionic Liquid and Hydroxypropyl-β-cyclodextrin for the Enantioseparation of Ten Drugs by Capillary Electrophoresis

In the present study, hydroxypropyl-β-cyclodextrin and an ionic liquid (1-ethyl-3-methylimidazolium-l-lactate) were used as additives in capillary electrophoresis for the enantioseparation of 10 analytes, including ofloxacin, propranolol hydrochloride, dioxopromethazine hydrochloride, isoprenaline hydrochloride, chlorpheniramine maleate, liarozole, tropicamide, amlodipine benzenesulfonate, brompheniramine maleate, and homatropine methylbromide. The effects of ionic liquid concentrations, salt effect, cations, and anions of ionic liquids on enantioseparation were investigated and the results proved that there was a synergistic effect between hydroxypropyl-β-cyclodextrin and the ionic liquid, and the cationic part of the ionic liquid played an important role in the increased resolution. With the developed dual system, all the enantiomers of 10 analytes were well separated in resolutions of 5.35, 1.76, 1.85, 2.48, 2.88, 1.43, 5.45, 4.35, 2.76, and 2.98, respectively. In addition, the proposed method was applied to the determination of the enantiomeric purity of S-ofloxacin after validation of the method in terms of selectivity, repeatability, linearity range, accuracy, precision, limit of detection (LOD), and limit of quality (LOQ).

[Prokaryotic Expression and Characterization of Two Recombinant Receptor-binding Domain(RBD) Proteins of Human Coronavirus NL63(HcoV-NL63)]

The receptor-binding domain(RBD) protein of HCoV-NL63 is a major target in the development of diagnostic assay and vaccine, it has a pivotal role in receptor attachment, viral entry and membrane fusion. In this study, we prepared 2 purified recombinant HCoV-NL63 RBD proteins using in E. coli system and identified the proteins by Western blotting. We first optimized codon and synthesized the RL (232-684aa)coding gene, then amplified the RL or RS(476-616aa) coding gene via PCR using different primers . The RL or RS coding gene was cloned into the pM48 expression vector fused with TrxA tag. The RBD (RL and RS) of HCoV-NL63 were expressed majorly as inclusion body when expressed in E. coli BL21pLys S under different conditions. The expressed products were purified by affinity chromatography then analyzed by SDS-PAGE and Western blotting. Our results showed that the recombinant RBD proteins were maximally expressed at 37 degrees C with 0. 8mM IPTG induction for 4h. RL or RS protein with 95 % purity was obtained and reacted positively with anti-sera from mice immunized with the recombinant vaccinia virus (Tiantan strain) in which HCoV-NL63 RL or RS protein was expressed. In conclusion, the purified recombinant RBD proteins(RL and RS)derived from E. coli were first prepared in China and they might provide a basis for further exploring biological role and vaccine development of HCoV-NL63.

Engineering Chiral Polyoxometalate Hybrid Metal-organic Frameworks for Asymmetric Dihydroxylation of Olefins

Chiral metal-organic frameworks (MOFs) with porous and tunable natures have made them feasible for performing a variety of chemical reactions as heterogeneous asymmetric catalysts. By incorporating the oxidation catalyst [BW12O40](5-) and the chiral group, L- or D-pyrrolidin-2-ylimidazole (PYI), into one single framework, the two enantiomorphs Ni-PYI1 and Ni-PYI2 were obtained via self-assembly, respectively. The channels of Ni-PYIs were enlarged through a guest exchange reaction to remove the cationic chiral templates and were well modulated with hydrophilic/hydrophobic properties to allow molecules of both H2O2 and olefin ingress and egress. The coexistence of both the chiral directors and the oxidants within a confined space provided a special environment for the formation of reaction intermediates in a stereoselective fashion with high selectivity. The resulting MOF acted as an amphipathic catalyst to prompt the asymmetric dihydroxylation of aryl olefins with excellent stereoselectivity.

HtrA3 is Negatively Correlated with Lymph Node Metastasis in Invasive Ductal Breast Cancer

Breast cancer is the most common cancer in women worldwide. Studies have shown that the high temperature requirement factor A3 (HtrA3) is involved in important physiological processes including maintenance of mitochondrial homeostasis, cell death, and cell signaling. HtrA3 is reported to be downregulated in several cancers and has been correlated with advancing cancer stage. We performed a retrospective study using our breast cancer tissue bank to investigate whether the expression of HtrA3 correlated with lymphatic metastasis in breast cancer and whether the expression of HtrA3 was correlated with estrogen receptor (ER) and progesterone receptor (PR) expression in breast cancer. Breast cancer tissues from 156 invasive ductal breast cancer patients with or without lymphatic metastasis were collected and the levels of HtrA3 were measured by immunohistochemistry and western blotting. The expression of HtrA3 was lower in breast cancer. In particular, HtrA3 expression in breast cancer with lymphatic metastasis was lower than that in breast cancer without lymphatic metastasis. In breast cancers with no lymphatic metastasis, the expression of HtrA3 was lower in patients with ER- and PR-positive tumors, but when breast cancers with lymphatic metastasis were analyzed, there was no difference in HtrA3 expression between ER- and PR-positive or ER- and PR-negative tumors. These data suggest that the expression of HtrA3 was negatively correlated with lymphatic metastasis in breast cancer but not correlated with ER and PR positivity or negativity. A better understanding of the mechanism of HtrA3 may provide the basis for future development of a novel therapeutic target in breast cancer.

Inhibition of Pathological Retinal Neovascularization by Semaphorin 3A

Pathological retinal angiogenesis is a major cause of vision loss. Semaphorin 3A (Sema3A), a chemorepellent guidance protein, plays crucial roles in neural and vascular patterning. To identify its role in retinal neovascularization, we investigated its antiangiogenic effects.

Differences in Platelet Indices Between Healthy Han Population and Tibetans in China

The present data on the evaluation of platelet (PLT) parameters in Chinese Han population and Tibetans are still limited. The objective of this study was to determine the differences in common PLT indices between Han population and Tibetans in China, through a large-scale investigation of healthy people.

The Effectiveness Comparison of Jitai Tablets Versus Methadone in Community-based Drug Treatment: a 1-year Follow-up Study

The aim of the study was to compare the effectiveness of Jitai tablets (JTT) versus methadone in a community drug treatment program.

A Signal Amplification Strategy Using the Cascade Catalysis of Gold Nanoclusters and Glucose Dehydrogenase for Ultrasensitive Detection of Thrombin

This work reports a novel signal amplification strategy for ultrasensitive detection of thrombin by cascade catalysis of gold nanoclusters (AuNCs) and glucose dehydrogenase (GDH). Herein, the AuNCs prepared by using polyamidoamine dendrimer as template were constructed not only as nanocarriers for anchoring the large amounts of secondary thrombin aptamers but also as nanocatalysts to catalyze the oxidation of NADH efficiently. Moreover, a large amount of GDH was loaded through the immobilization technology of DNA hybridization and a large amount of toluidine blue (Tb) was intercalated into the DNA grooves via electrostatic interaction. Significantly, the electrochemical signal was greatly enhanced based on cascade catalysis: firstly, GDH catalyzed the oxidation of glucose to gluconolactone with the concomitant generation of NADH in the presence of NAD(+). Then, AuNCs as nanocatalysts could effectively catalyze NADH to produce NAD(+) with the help of Tb as redox probe. Under the optimal conditions, the proposed aptasensor exhibits a linear range of 1.0×10(-14)-5×10(-9) M with a low detection limit of 3.3×10(-15) M for thrombin detection and shows high sensitivity and good specificity.

Hepatitis C Virus Infection is Independently Associated with Depression Among Methadone Maintenance Treatment Heroin Users in China

Depression and hepatitis C virus (HCV) infection are two common conditions among heroin users in methadone maintenance treatment (MMT). However, the comorbid relationship between depression and HCV infection among MMT patients is not well understood.

Teachers of Psychiatry Meeting on Evaluation to Improve Quality of Research, Held in Shanghai, China, Organized by the Pacific Rim College of Psychiatrists

Histone Demethylase Retinoblastoma Binding Protein 2 is Overexpressed in Hepatocellular Carcinoma and Negatively Regulated by Hsa-miR-212

The H3K4 demethylase retinoblastoma binding protein 2 (RBP2) is involved in the pathogenesis of gastric cancer, but its role and regulation in hepatocellular carcinoma (HCC) is unknown. We determined the function of RBP2 and its regulation in HCC in vitro and in human tissues.

Safety and Immunological Responses to Human Mesenchymal Stem Cell Therapy in Difficult-to-treat HIV-1-infected Patients

HAART largely decreases morbidity and mortality in chronic HIV-1-infected patients, but immune nonresponders (INRs) with full viral suppression still fail to reverse the immune deficiency. This study evaluated the safety and immunological responses of human umbilical cord mesenchymal stem cell (MSC) therapy in HIV-1-infected INRs.

Inhibitory Effects of Rosiglitazone on Paraquat-induced Acute Lung Injury in Rats

To investigate the effects of the PPAR-γ agonist rosiglitazone on acute lung injury induced by the herbicide paraquat (PQ) and the underlying mechanisms of action.

Expression and Correlation of Twist and Gelatinases in Breast Cancer

Altered expression of Twist, matrix metalloproteinase (MMP)-2 and MMP-9 proteins has been identified in various types of human cancers. However, the correlation between Twist and these gelatinases in breast cancer remains unclear. In this study, immunohistochemical analysis of Twist, MMP-2 and MMP-9 expression was performed on tissue microarrays from 200 breast cancer cases. The association of Twist and gelatinase expression with clinicopathological factors and patient survival was analyzed. Altered expression of Twist, MMP-2 and MMP-9 proteins was observed in breast cancer tissue. The positive rates of Twist, MMP-2 and MMP-9 protein expression were 75.5, 97.0 and 96.0%, respectively. Increased expression of Twist was positively correlated with the status of axillary lymph node metastasis and higher tumor-node-metastasis (TNM) stage (P<0.01). Moreover, increased expression of Twist was correlated with poor overall survival (OS) and post-operative relapse-free survival (RFS), compared with those for the patients with reduced expression levels of Twist (P<0.05, P<0.01). The expression of MMP-2 and MMP-9 was positively correlated with Twist expression (P<0.001). Our results indicate that Twist may play an important role in the invasion, metastasis and prognosis of breast cancer. Additionally, our results suggest that Twist may be a regulator of gelatinases (MMP-2 and MMP-9).

MMP-7 and TIMP-1, New Targets in Predicting Poor Wound Healing in Apical Periodontitis

Matrix metalloproteinases (MMPs) and the tissue inhibitors of metalloproteinases (TIMPs) are strongly associated with tissue destruction because of inflammation. In this study, we investigated the expression of MMPs and TIMPs messenger RNA and protein levels in apical periodontitis lesions.

An Analysis of the Indications for Cesarean Section in a Teaching Hospital in China

Cesarean delivery rates have increased remarkably worldwide. The indications for this increase are not fully understood and there may be regional, ethnic or health system differences in quoted indications which may explain, at least in part, the observed changes. In 2008 China was cited as having one of the highest rates of cesarean delivery in the world, but there was no accurate information about the indications for the high rate. This study sought to provide some information about the high cesarean section rate in China.

Short-term Clinical Effects of Photodynamic Therapy with Topical 5-aminolevulinic Acid for Facial Acne Conglobata: an Open, Prospective, Parallel-arm Trial

Acne conglobata is hardly curable and easily leads to scar formation after treatment using traditional methods.

KH902, a Recombinant Human VEGF Receptor Fusion Protein, Reduced the Level of Placental Growth Factor in Alkali Burn Induced-corneal Neovascularization

To investigate the expression of placental growth factor (PIGF) in alkali burn-induced murine corneal neovascularization (NV); to evaluate the effects of KH902, a vascular endothelial growth factor receptor decoy, on prevention and regression of new vessels growths in the cornea; and to determine the influence of KH902 on the levels of vascular endothelial growth factor (VEGF) and PIGF in alkali burn-induced corneal NV.

Reliability and Validity of the CogState Battery Chinese Language Version in Schizophrenia

Cognitive impairment in patients with schizophrenia is a core symptom of this disease. The computerized CogState Battery (CSB) has been used to detect seven of the most common cognitive domains in schizophrenia. The aim of this study was to examine the reliability and validity of the Chinese version of the CSB (CSB-C), in Chinese patients with schizophrenia.

IQdb: an Intelligence Quotient Score-associated Gene Resource for Human Intelligence

Intelligence quotient (IQ) is the most widely used phenotype to characterize human cognitive abilities. Recent advances in studies on human intelligence have identified many new susceptibility genes. However, the genetic mechanisms involved in IQ score and the relationship between IQ score and the risk of mental disorders have won little attention. To address the genetic complexity of IQ score, we have developed IQdb (http://IQdb.cbi.pku.edu.cn), a publicly available database for exploring IQ-associated human genes. In total, we collected 158 experimental verified genes from literature as a core dataset in IQdb. In addition, 46 genomic regions related to IQ score have been curated from literature. Based on the core dataset and 46 confirmed linked genomic regions, more than 6932 potential IQ-related genes are expanded using data of protein-protein interactions. A systematic gene ranking approach was applied to all the collected and expanded genes to represent the relative importance of all the 7090 genes in IQdb. Our further systematic pathway analysis reveals that IQ-associated genes are significantly enriched in multiple signal events, especially related to cognitive systems. Of the 158 genes in the core dataset, 81 are involved in various psychotic and mental disorders. This comprehensive gene resource illustrates the importance of IQdb to our understanding on human intelligence, and highlights the utility of IQdb for elucidating the functions of IQ-associated genes and the cross-talk mechanisms among cognition-related pathways in some mental disorders for community. Database URL: http://IQdb.cbi.pku.edu.cn.

Dual Signal Amplification Strategy for the Fabrication of an Ultrasensitive Electrochemiluminescenct Aptasensor

A dual signal amplification strategy was designed to construct a cathodic peroxydisulfate-based electrochemiluminescence (ECL) aptasensor for the ultrasensitive detection of thrombin (TB) as a model analyte. The signal was amplified by the employment of two kinds of co-reactant, (1) in situ generated dissolved O2 from the cascade catalysis of GOD and HRP as co-reactants and (2) intercalation of a new co-reactant into the grooves of the dsDNA polymers based on the hybridization chain reaction (HCR). Femtomolar levels of TB could be detected using the developed aptasensor, with high sensitivity and good stability. Thus the S2O8(2-)-O2 ECL system has great potential for development and application in clinical diagnostics, immunology and biomedical research.

Effects of Semaphorin 3A on Retinal Pigment Epithelial Cell Activity

Semaphorin 3A (Sema3A), a chemorepellant guidance protein, has been shown to be crucial for neural and vascular remodeling. This study is designed to examine the effects of Sema3A on RPE cell activity both in vitro and in vivo.

First Definition of Reference Intervals of Liver Function Tests in China: a Large-population-based Multi-center Study About Healthy Adults

Reference intervals of Liver function tests are very important for the screening, diagnosis, treatment, and monitoring of liver diseases. We aim to establish common reference intervals of liver function tests specifically for the Chinese adult population.

[Anatomy Study on Micro Transverse Flap Pedicled with Superfical Palmar Branch of Radial Artery from Palmar Wrist]

To study the anatomical basis of micro transverse flap pedicled with the superfical palmar branch of radial artery from the palmar wrist for using this free flap to repair soft tissue defect of the finger.

Characterization of Schizophrenia Adverse Drug Interactions Through a Network Approach and Drug Classification

Antipsychotic drugs are medications commonly for schizophrenia (SCZ) treatment, which include two groups: typical and atypical. SCZ patients have multiple comorbidities, and the coadministration of drugs is quite common. This may result in adverse drug-drug interactions, which are events that occur when the effect of a drug is altered by the coadministration of another drug. Therefore, it is important to provide a comprehensive view of these interactions for further coadministration improvement. Here, we extracted SCZ drugs and their adverse drug interactions from the DrugBank and compiled a SCZ-specific adverse drug interaction network. This network included 28 SCZ drugs, 241 non-SCZs, and 991 interactions. By integrating the Anatomical Therapeutic Chemical (ATC) classification with the network analysis, we characterized those interactions. Our results indicated that SCZ drugs tended to have more adverse drug interactions than other drugs. Furthermore, SCZ typical drugs had significant interactions with drugs of the "alimentary tract and metabolism" category while SCZ atypical drugs had significant interactions with drugs of the categories "nervous system" and "antiinfectives for systemic uses." This study is the first to characterize the adverse drug interactions in the course of SCZ treatment and might provide useful information for the future SCZ treatment.

Synergetic Regulatory Networks Mediated by Oncogene-driven MicroRNAs and Transcription Factors in Serous Ovarian Cancer

Although high-grade serous ovarian cancer (OVC) is the most lethal gynecologic malignancy in women, little is known about the regulatory mechanisms in the cellular processes that lead to this cancer. Recently, accumulated lines of evidence have shown that the interplay between transcription factors (TFs) and microRNAs (miRNAs) is critical in cellular regulation during tumorigenesis. A comprehensive investigation of TFs and miRNAs, and their target genes, may provide a deeper understanding of the regulatory mechanisms in the pathology of OVC. In this study, we have integrated three complementary algorithms into a framework, aiming to infer the regulation by miRNAs and TFs in conjunction with gene expression profiles. We demonstrated the utility of our framework by inferring 67 OVC-specific regulatory feed-forward loops (FFL) initiated by miRNAs or TFs in high-grade serous OVC. By analyzing these regulatory behaviors, we found that all the 67 FFLs are consistent in their regulatory effects on genes that are jointly targeted by miRNAs and TFs. Remarkably, we unveiled an unbalanced distribution of FFLs with different oncogenic effects. In total, 31 of the 67 coherent FFLs were mainly initiated by oncogenes. On the contrary, only 4 of the FFLs were initiated by tumor suppressor genes. These overwhelmingly observed oncogenic genes were further detected in a sub-network with 32 FFLs centered by miRNA let-7b and TF TCF7L1 to regulate cell differentiation. Closer inspection of 32 FFLs revealed that 75% of the miRNAs reportedly play functional roles in cell differentiation, especially when enriched in epithelial-mesenchymal transitions. This study provides a comprehensive pathophysiological overview of recurring coherent circuits in OVC that are co-regulated by miRNAs and TFs. The prevalence of oncogenic coherent FFLs in serous OVC suggests that oncogene-driven regulatory motifs could cooperatively act upon critical cellular processes such as cell differentiation in a highly efficient and consistent manner.

TOMM40 Rs2075650 Polymorphism Shows No Association with Neovascular Age-related Macular Degeneration or Polypoidal Choroidal Vasculopathy in a Chinese Population

Age-related macular degeneration (AMD) and Alzheimer disease (AD) are age-related neurodegenerative diseases that share similar environmental risk factors, cellular pathologies, and genetic backgrounds. Recently, the rs2075650 single nucleotide polymorphism in the translocase of outer mitochondrial membrane 40 homolog (TOMM40) gene was identified as a risk factor for AMD and Alzheimer disease. We aimed to examine the associations between the TOMM40 rs2075650 polymorphism and neovascular age-related macular degeneration (nAMD) and polypoidal choroidal vasculopathy (PCV) in a Chinese population.

Evolutionary and Ontogenetic Changes in RNA Editing in Human, Chimpanzee, and Macaque Brains

Adenosine-to-inosine (A-to-I) substitutions are the most common type of RNA editing in mammals. A-to-I RNA editing is particularly widespread in the brain and is known to play important roles in neuronal functions. In this study we investigated RNA-editing changes during human brain development and maturation, as well as evolutionary conservation of RNA-editing patterns across primates. We used high-throughput transcriptome sequencing (RNA-seq) to quantify the RNA-editing levels and assess ontogenetic dynamics of RNA editing at more than 8000 previously annotated exonic A-to-I RNA-editing sites in two brain regions--prefrontal cortex and cerebellum--of humans, chimpanzees, and rhesus macaques. We observed substantial conservation of RNA-editing levels between the brain regions, as well as among the three primate species. Evolutionary changes in RNA editing were nonetheless evident, with 40% of the annotated editing sites studied showing divergent editing levels among the three species and 16.5% of sites displaying statistically significant human-specific editing patterns. Across lifespan, we observed an increase of the RNA-editing level with advanced age in both brain regions of all three primate species.

Characterization of Two Distinct Neuraminidases from Avian-origin Human-infecting H7N9 Influenza Viruses

An epidemic of an avian-origin H7N9 influenza virus has recently emerged in China, infecting 134 patients of which 45 have died. This is the first time that an influenza virus harboring an N9 serotype neuraminidase (NA) has been known to infect humans. H7N9 viruses are divergent and at least two distinct NAs and hemagglutinins (HAs) have been found, respectively, from clinical isolates. The prototypes of these viruses are A/Anhui/1/2013 and A/Shanghai/1/2013. NAs from these two viruses are distinct as the A/Shanghai/1/2013 NA has an R294K substitution that can confer NA inhibitor oseltamivir resistance. Oseltamivir is by far the most commonly used anti-influenza drug due to its potency and high bioavailability. In this study, we show that an R294K substitution results in multidrug resistance with extreme oseltamivir resistance (over 100 000-fold) using protein- and virus-based assays. To determine the molecular basis for the inhibitor resistance, we solved high-resolution crystal structures of NAs from A/Anhui/1/2013 N9 (R294-containing) and A/Shanghai/1/2013 N9 (K294-containing). R294K substitution results in an unfavorable E276 conformation for oseltamivir binding, and consequently loss of inhibitor carboxylate interactions, which compromises the binding of all classical NA ligands/inhibitors. Moreover, we found that R294K substitution results in reduced NA catalytic efficiency along with lower viral fitness. This helps to explain why K294 has predominantly been found in clinical cases of H7N9 infection under the selective pressure of oseltamivir treatment and not in the dominant human-infecting viruses. This implies that oseltamivir can still be efficiently used in the treatment of H7N9 infections.

Characterization of a Soluble B7-H3 (sB7-H3) Spliced from the Intron and Analysis of SB7-H3 in the Sera of Patients with Hepatocellular Carcinoma

B7-H3 is a recently discovered member of the B7 superfamily molecules and has been found to play a negative role in T cell responses. In this study, we identified a new B7-H3 isoform that is produced by alternative splicing from the forth intron of B7-H3 and encodes the sB7-H3 protein. Protein sequence analysis showed that sB7-H3 contains an additional four amino acids, encoded by the intron sequence, at the C-terminus compared to the ectodomain of 2Ig-B7-H3. We further found that this spliced sB7-H3 plays a negative regulatory role in T cell responses and serum sB7-H3 is higher in patients with hepatocellular carcinoma (HCC) than in healthy donors. Furthermore, we found that the expression of the spliced sb7-h3 gene is higher in carcinoma and peritumor tissues than in PBMCs of both healthy controls and patients, indicating that the high level of serum sB7-H3 in patients with HCC is caused by the increased expression of this newly discovered spliced sB7-H3 isoform in carcinoma and peritumor tissues.

Effects of Prunella Vulgaris on the Mice Immune Function

The present study was designed to evaluate the effects of Prunella Vulgaris (P. vulgaris) on the immune function in mice. The mice were randomly divided into one control group and three treatment groups of 10 mice each. The control group received pure water and the treatment groups received P. vulgaris extract at concentrations of 0.15, 0.30 and 0.90 g/kg BW orally for 30 days, respectively. Changes in cell immune function, non-specific immunity and humoral immunity function were evaluated. Active lymphocytes and T lymphocyte subsets were determined by fluorescence-activated cell sorting (FACS). Certain Serum concentrations of cytokines were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that, for cell immune function, compared with the control group, foot pad thickness in high dose group increased significantly (p<0.01), whereas no significant difference in the proliferative ability of splenic lymphocytes was observed among all groups (p>0.05). For non-specific immunity, NK cell activity increased significantly in a dose-dependent manner in P. vulgaris treated mice (p<0.01), mononuclear-macrophage function in medium and high dose P. vulgaris treated mice were significantly higher than that of the control group (p<0.05). For humoral immunity, no significant differences were observed in terms of the half value of hemolysis (HC50), number of hemolytic plaques and serum IgG level (p>0.05). The percentage of active T and Th lymphocytes of mice peripheral blood in high dose group were significantly higher than that of the control group (p<0.01). There was no significant difference in serum levels of IL-1β, IL-4, IL-10 and IFN-γ among all of the four groups (p>0.05). The data indicated that 0.90 g/kg BW P. vulgaris extract (equivalent to 7.5 g/kg BW crude drug) had some effect on cellular immune function and non-specific immune function in mice.

Differential Responses of Hepatic Endoplasmic Reticulum Stress and Inflammation in Diet-induced Obese Rats with High-fat Diet Rich in Lard Oil or Soybean Oil

To investigate the effects of high-fat diet enriched with lard oil or soybean oil on liver endoplasmic reticulum (ER) stress and inflammation markers in diet-induced obese (DIO) rats and estimate the influence of following low-fat diet feeding.

Metabonomics Study of the Therapeutic Mechanism of Gynostemma Pentaphyllum and Atorvastatin for Hyperlipidemia in Rats

Gynostemma pentaphyllum (GP) is widely used for the treatment of diseases such as hyperlipidemia, fatty liver and obesity in China, and atorvastatin is broadly used as an anti-hyperlipidemia drug. This research focuses on the plasma and liver metabolites in the following four groups of rats: control, a hyperlipidemia model, a hyperlipidemia model treated with GP and a hyperlipidemia model treated with atorvastatin. Using (1)H-NMR-based metabonomics, we elucidated the therapeutic mechanisms of GP and atorvastatin. Orthogonal Partial Least Squares-Discriminant analysis (OPLS-DA) plotting of the metabolic state and analysis of potential biomarkers in the plasma and liver correlated well with the results of biochemical assays. GP can effectively affect lipid metabolism, and it exerts its anti-hyperlipidemia effect by elevating the level of phosphatidylcholine and decreasing the level of trimethylamine N-oxide (TMAO). In contrast, atorvastatin affects hyperlipidemia mainly during lipid metabolism and protein metabolism in vivo.

TNFRSF10A-LOC389641 Rs13278062 but Not REST-C4orf14-POLR2B-IGFBP7 Rs1713985 Was Found Associated with Age-related Macular Degeneration in a Chinese Population

To reassess the association between TNFRSF10-LOC389641 rs13278062 and REST-C4orf14-POLR2B-IGFBP7 rs1713985 with the risk of AMD in a Chinese case-control collection.

Generation of a Tenascin-C-CreER2 Knockin Mouse Line for Conditional DNA Recombination in Renal Medullary Interstitial Cells

Renal medullary interstitial cells (RMIC) are specialized fibroblast-like cells that exert important functions in maintaining body fluid homeostasis and systemic blood pressure. Here, we generated a RMIC specific tenascin-C promoter driven inducible CreER2 knockin mouse line with an EGFP reporter. Similar as endogenous tenascin-C expression, the reporter EGFP expression in the tenascin-C-CreER2(+/-) mice was observed in the inner medulla of the kidney, and co-localized with COX2 but not with AQP2 or AQP1, suggesting selective expression in RMICs. After recombination (tenascin-C-CreER2(+/-)/ROSA26-lacZ(+/-) mice + tamoxifen), β-gal activity was restricted to the cells in the inner medulla of the kidney, and didn't co-localize with AQP2, consistent with selective Cre recombinase activity in RMICs. Cre activity was not obvious in other major organs or without tamoxifen treatment. This inducible RMIC specific Cre mouse line should therefore provide a novel tool to manipulate genes of interest in RMICs.

CNVannotator: a Comprehensive Annotation Server for Copy Number Variation in the Human Genome

Copy number variation (CNV) is one of the most prevalent genetic variations in the genome, leading to an abnormal number of copies of moderate to large genomic regions. High-throughput technologies such as next-generation sequencing often identify thousands of CNVs involved in biological or pathological processes. Despite the growing demand to filter and classify CNVs by factors such as frequency in population, biological features, and function, surprisingly, no online web server for CNV annotations has been made available to the research community. Here, we present CNVannotator, a web server that accepts an input set of human genomic positions in a user-friendly tabular format. CNVannotator can perform genomic overlaps of the input coordinates using various functional features, including a list of the reported 356,817 common CNVs, 181,261 disease CNVs, as well as, 140,342 SNPs from genome-wide association studies. In addition, CNVannotator incorporates 2,211,468 genomic features, including ENCODE regulatory elements, cytoband, segmental duplication, genome fragile site, pseudogene, promoter, enhancer, CpG island, and methylation site. For cancer research community users, CNVannotator can apply various filters to retrieve a subgroup of CNVs pinpointed in hundreds of tumor suppressor genes and oncogenes. In total, 5,277,234 unique genomic coordinates with functional features are available to generate an output in a plain text format that is free to download. In summary, we provide a comprehensive web resource for human CNVs. The annotated results along with the server can be accessed at http://bioinfo.mc.vanderbilt.edu/CNVannotator/.

Flexible Free-standing Luminescent Two-component Fiber Films with Tunable Hierarchical Structures Based on Hydrogen-bonding Architecture

Although the fabrication of hierarchical architectures with highly ordered functional units is of great importance for both fundamental science and practical application, the development of one-dimensional (1D) organic hierarchical micro/nanostructures based on low-molecular-weight (LMW) building blocks remains at an early stage. Herein, we report two types of micro/nanoscaled multicomponent fluorescent fiber systems with tunable hierarchical morphologies through a one-step coassembly process. With the aid of hydrogen-bonding motifs, LMW precursors (1,4-bis(5-phenyloxazol-2-yl)benzene (A) and two coassembled building blocks: 4-bromotetrafluorobenzene carboxylic acid (B) and 2,3,4,5,6-pentafluorophenol (C)) have been self-organized into fibers and flexible free-standing films, which show hierarchical micro/nanostructures as well as tunable one-/two-photon luminescence. The disassembly of the multicomponent A.B and A.C fibers occurs at high temperature, which further alters the luminescence properties of the multicomponent materials. Therefore, this work provides a facile wet chemical route for fabricating multicomponent LMW self-assembled fibers and free-standing film systems with tunable hierarchical structures and photoemission behaviors, and such self-organized systems may have potential applications in fields of two-photon luminescence and thermal sensors.

EDdb: a Web Resource for Eating Disorder and Its Application to Identify an Extended Adipocytokine Signaling Pathway Related to Eating Disorder

Eating disorder is a group of physiological and psychological disorders affecting approximately 1% of the female population worldwide. Although the genetic epidemiology of eating disorder is becoming increasingly clear with accumulated studies, the underlying molecular mechanisms are still unclear. Recently, integration of various high-throughput data expanded the range of candidate genes and started to generate hypotheses for understanding potential pathogenesis in complex diseases. This article presents EDdb (Eating Disorder database), the first evidence-based gene resource for eating disorder. Fifty-nine experimentally validated genes from the literature in relation to eating disorder were collected as the core dataset. Another four datasets with 2824 candidate genes across 601 genome regions were expanded based on the core dataset using different criteria (e.g., protein-protein interactions, shared cytobands, and related complex diseases). Based on human protein-protein interaction data, we reconstructed a potential molecular sub-network related to eating disorder. Furthermore, with an integrative pathway enrichment analysis of genes in EDdb, we identified an extended adipocytokine signaling pathway in eating disorder. Three genes in EDdb (ADIPO (adiponectin), TNF (tumor necrosis factor) and NR3C1 (nuclear receptor subfamily 3, group C, member 1)) link the KEGG (Kyoto Encyclopedia of Genes and Genomes) "adipocytokine signaling pathway" with the BioCarta "visceral fat deposits and the metabolic syndrome" pathway to form a joint pathway. In total, the joint pathway contains 43 genes, among which 39 genes are related to eating disorder. As the first comprehensive gene resource for eating disorder, EDdb ( http://eddb.cbi.pku.edu.cn ) enables the exploration of gene-disease relationships and cross-talk mechanisms between related disorders. Through pathway statistical studies, we revealed that abnormal body weight caused by eating disorder and obesity may both be related to dysregulation of the novel joint pathway of adipocytokine signaling. In addition, this joint pathway may be the common pathway for body weight regulation in complex human diseases related to unhealthy lifestyle.

Four New 7,8-epoxycembranoids from a Chinese Soft Coral Lobophytum Sp

Four new 7,8-epoxycembranoids, namely (2S*,7S*,8S*,12R*,1Z,3E,10E)-7,8:2,16-diepoxycembra-1(15),3,10-trien-12-ol (1), (2S*,7S*,8S*,11R*,1Z,3E)-7,8:2,16-diepoxycembra-1(15),3,12(20)-trien-11-ol (2), (4S*,7S*,8S*,1Z,2E,11E)-16-acetoxy-7,8-epoxycembra-1(15),2,11-trien-4-ol (3), and (7S*,8S*,15S*,1E,3E,11E)-7,8-epoxycembra-1,3,11-trien-15,16-diol (4) were isolated from a Chinese soft coral Lobophytum sp., together with eleven known analogues 5-15. The structures of the new compounds were determined by extensive spectroscopic data analysis. All compounds were tested for the inhibitory effect on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in mouse peritoneal macrophages (PEMΦ).

[Analysis of Clinical Effects of Parenterally Administered Shenqi Fuzheng on Renal Function]

This study analyzes the clinical effects of parenterally administered Shenqi Fuzheng on renal function. 20 national, general hospitals were selected. Their hospital information system (HIS) data on 51 898 cases of parenterally administered Shenqi Fuzheng were mined for data. Patients ranged from 18 to 80 years old. 27 718 cases were selected for analysis. Serum creatinine (Scr) and blood urea nitrogen (BUN) levels were taken before and after treatment for outcome evaluation. According to instructions, we divided 197 cases into the treatment group (doses > 250 mL) and 5 728 cases acted as the control group (dose < or = 250 mL). Stratified analysis adjusted for age, sex, hospital illness, treatment, etc. According to the four variables, the case group compared with the control group did not show abnormal renal function changes; 57 confounding factors were balanced using propensity score method resulting in the treatment group showing no abnormal changes in renal function. This HIS b data analysis found that parenterally administered Shenqi Fuzheng above the recommended dosage did not significantly impact on renal function was no significant difference. Prospective studies should be carried out to validate this data.

[Application of Immunohistochemistry in Diagnosis of ALK-positive Non-small Cell Lung Cancer]

Computational Tools for Copy Number Variation (CNV) Detection Using Next-generation Sequencing Data: Features and Perspectives

Copy number variation (CNV) is a prevalent form of critical genetic variation that leads to an abnormal number of copies of large genomic regions in a cell. Microarray-based comparative genome hybridization (arrayCGH) or genotyping arrays have been standard technologies to detect large regions subject to copy number changes in genomes until most recently high-resolution sequence data can be analyzed by next-generation sequencing (NGS). During the last several years, NGS-based analysis has been widely applied to identify CNVs in both healthy and diseased individuals. Correspondingly, the strong demand for NGS-based CNV analyses has fuelled development of numerous computational methods and tools for CNV detection. In this article, we review the recent advances in computational methods pertaining to CNV detection using whole genome and whole exome sequencing data. Additionally, we discuss their strengths and weaknesses and suggest directions for future development.

[Risk Factors Analysis of Local Failure Following Radiotherapy and Chemotherapy to Nasopharyngeal Carcinoma]

To analyse the risk factors involved in local failure following radiotherapy and chemotherapy to nasopharyngeal carcinoma.

[Clinical Application of Micro Transverse Flap Pedicled with Superficial Palmar Branch of Radial Artery from Palmar Wrist to Repair Skin Defect of Finger]

To investigate the clinical application of micro transverse flap pedicled with superficial palmar branch of radial artery from palmar wrist to repair skin defect of finger.

Case-control Study of Error-related Negativity Among Males with Heroin Dependence Undergoing Rehabilitation

There were 1.2 million registered heroin users in China by the end of 2011, but little research in the country has focused on the neuropsychological functioning of these individuals.

Down-regulation of MicroRNA-29c is Associated with Renal Failure in Multiple Myeloma

Increased Dietary Sodium Induces COX2 Expression by Activating NFκB in Renal Medullary Interstitial Cells

High salt diet induces renal medullary cyclooxygenase 2 (COX2) expression. Selective blockade of renal medullary COX2 activity in rats causes salt-sensitive hypertension, suggesting a role for renal medullary COX2 in maintaining systemic sodium balance. The present study characterized the cellular location of COX2 induction in the kidney of mice following high salt diet and examined the role of NFκB in mediating this COX2 induction in response to increased dietary salt. High salt diet (8 % NaCl) for 3 days markedly increased renal medullary COX2 expression in C57Bl/6 J mice. Co-immunofluorescence using a COX2 antibody and antibodies against aquaporin-2, ClC-K, aquaporin-1, and CD31 showed that high salt diet-induced COX2 was selectively expressed in renal medullary interstitial cells. By using NFκB reporter transgenic mice, we observed a sevenfold increase of luciferase activity in the renal medulla of the NFκB-luciferase reporter mice following high salt diet, and a robust induction of enhanced green fluorescent protein (EGFP) expression mainly in renal medullary interstitial cells of the NFκB-EGFP reporter mice following high salt diet. Treating high salt diet-fed C57Bl/6 J mice with selective IκB kinase inhibitor IMD-0354 (8 mg/kg bw) substantially suppressed COX2 induction in renal medulla, and also significantly reduced urinary prostaglandin E2 (PGE2). These data therefore suggest that renal medullary interstitial cell NFκB plays an important role in mediating renal medullary COX2 expression and promoting renal PGE2 synthesis in response to increased dietary sodium.

Pan-amyloid Oligomer Specific ScFv Antibody Attenuates Memory Deficits and Brain Amyloid Burden in Mice with Alzheimer's Disease

Amyloid oligomers have a critical function in the pathologic processes of various amyloidoses, such as Alzheimer's disease (AD), Parkinson disease (PD), Huntington's disease, prion-related diseases, type 2 diabetes, and hereditary renal amyloidosis. Our previous reports demonstrated that a conformation-dependent oligomer-specific single-chain variable fragment (scFv) antibody, W20, isolated from a naïve human scFv library, can recognize oligomers assembled from α-synuclein, amylin, insulin, Aβ40/42, prion peptide 106-126, and lysozyme, inhibit the aggregation of various amyloid, and attenuate amyloid oligomer-induced cytotoxicity In vitro. Furthermore, W20 recognized the amyloid oligomers in all types of plaques, Lewy bodies, and amylin deposits in the brain tissues of AD and PD patients and in the pancreas of type 2 diabetes patients. In the current study, we showed that W20 blocked the binding of Aβ oligomers to SH-SY5Y cells, did not bind to heat shock protein, rescued cognitive impairments in APP/PS1 transgenic mice, and interfered with Aβ levels and deposits in mouse brain. These results suggest that W20 may be a promising therapeutic for the treatment of AD.

Combined Use of Hydroxypropyl-β-cyclodextrin and Ionic Liquids for the Simultaneous Enantioseparation of Four Azole Antifungals by CE and a Study of the Synergistic Effect

A CE method employing a dual system of hydroxypropyl-β-cyclodextrin (HP-β-CD) and ionic liquids (ILs) has been developed for the simultaneous enantioseparation of four azole antifungals for the first time. In this study, three different types of ILs were employed as modifiers and among them dodecyl trimethyl ammonium chloride was found to be the most effective. The effects of the concentration, cations, and anions of ILs on the enantioseparation were investigated. With the developed dual system, all the enantiomers were well separated in resolutions of 3.8, 3.5, 2.8, and 2.5 for miconazole, econazole, ketoconazole, and itraconazole, respectively. The interactions between dodecyl trimethyl ammonium chloride and HP-β-CD were also studied using a neutral polyacrylamide coated capillary and (1) H NMR spectroscopy to further explore the synergistic effect involved. It was found that ILs improved the enantioseparation not only by changing the EOF, but also by interactions with HP-β-CD that could change its ability of forming inclusion complex with the enantiomers.

The Preparation of Gold Nanoparticles and Evaluation of Their Immunological Function Effects on Rats

As a new type of biomaterials, gold nanoparticles (GNPs), also known as colloidal gold (CG), have a wide biomedical application. In this study, GNPs with diameters of 10, 15, and 25 nm were prepared by sodium citrate reduction, and detected by common optical property, ultraviolet-visible (UV-vis) absorbance spectroscopy, and scanning electron microscope (SEM), separately for identification of the particle size and uniformity. In order to observe the effects of GNPs on immune function, adult Sprague Dawley (SD) rats were immunized with the above three GNPs, each having three doses of 0.2, 0.4, and 0.6 ml, and rats without immunization served as negative control. After immunization, proliferation activity of blood and spleen lymphocyte and the levels of interleukin-2 (IL-2) in serum and supernatant of spleen lymphocyte were detected by thiazoleblue (MTT) assay and enzyme linked immunosorbent assay (ELISA), respectively. The results indicated that different size of GNPs was prepared, and the uniformity increased with the decrease of the size of particles. Different diameters and doses of GNPs have different effects on proliferation of blood and spleen lymphocyte, as well as the levels of IL-2 in serum and supernatant of spleen lymphocyte. The 15 nm CG in 0.6 ml dose group could most significantly promote blood and spleen lymphocyte proliferation, and enhance IL-2 levels in serum and supernatant of spleen lymphocyte. Taken together, the findings revealed that application of CG prepared by sodium citrate reduction could enhance specific and nonspecific immune responses, and the 0.6 ml dose of 15 nm CG might be the best immunizing dose in rats. This fact may serve as a further evidence for using CG as a novel immunoadjuvant in the future.

Peritoneal Dialysis-related Peritonitis with Acinetobacter Baumannii: a Review of Seven Cases

Peritonitis is still known as an important complication of continuous ambulatory peritoneal dialysis (CAPD). Multi-drug resistant (MDR) Acinetobacter baumannii is an increasing problem worldwide. Moreover, the increasing reports of carbapenem-resistant A. baumannii strains is common. Although peritoneal dialysis-related peritonitis with MDR A. baumannii is rarely reported, infection with this organism always results in serious peritonitis and increases the possibility of dropout or mortality. Here, we present 7 cases of peritonitis caused by A. baumannii species. Among those 7 cases, 2 involved MDR A. baumannii, and 1 involved a carbapenem-resistant strain. All the MDR bacterial infections failed treatment. We also review the literature about Acinetobacter peritonitis and current treatment protocols.

Ultrasensitive Apurinic/apyrimidinic Endonuclease 1 Immunosensing Based on Self-enhanced Electrochemiluminescence of a Ru(II) Complex

An alternative "signal on" immunosensor for ultrasensitive detection of apurinic/apyrimidinic endonuclease 1 (APE-1) was designed utilizing the self-enhanced electrochemiluminescence (ECL) of a novel Ru(II) complex functionalized coil-like nanocomposite as signal labels. The desirable self-enhanced ECL luminophore was achieved by combining the coreactant of poly(ethylenimine) (PEI) and the luminophor of bis(2,2'-bipyridine)-5-amino-1,10-phenanthroline ruthenium(II) [Ru(bpy)2(5-NH2-1,10-phen)(2+)] to form one novel Ru(II) complex, which exhibited significantly enhanced ECL efficiency and stability. Moreover, the carbon nanotubes (CNTs) were employed as nanocarriers for self-enhanced Ru(II) complex loading via π-π stacking to obtain the coil-like nanocomposite to act as signal probe. Compared with traditional ECL immunoassay, our proposed strategy is simple and sensitive, avoiding the adding of any coreactant into testing solution for signal amplification, and shows a detection limit down to subfemtogram per milliliter level under the optimized experimental condition.

A Luminescent Dye@MOF Platform: Emission Fingerprint Relationships of Volatile Organic Molecules

Self-assembly of luminescent moieties into porous metal-organic frameworks (MOFs) has generated many luminescent platforms for probing volatile organic molecules (VOMs). However, most of those explored thus far have only been based on the luminescence intensity of one transition, which is not efficient for probing different VOMs. We have synthesized a luminescent MOF material containing 1D nanotube channels, and further developed a luminescent dye@MOF platform to realize the probing of different VOMs by tuning the energy transfer efficiency between two different emissions. The dye@MOF platform exhibits excellent fingerprint correlation between the VOM and the emission peak-height ratio of ligand to dye moieties. The dye@MOF sensor is self-calibrating, stable, and instantaneous, thus the approach should be a very promising strategy to develop luminescent materials with unprecedented practical applications.

Identification of Astilbin Metabolites Produced by Human Intestinal Bacteria Using UPLC-Q-TOF/MS

Astilbin, mainly isolated from a commonly used herbal medicine, Smilax glabra Roxb (SGR), exhibits a variety of pharmacological activities and biological effects. It is metabolized by intestinal bacteria after oral administration which leads to the variation of ethnopharmacological profile of this traditional medicine. However, little is known on the interactions of this active compound with intestinal bacteria, which would be very helpful in unravelling how SGR works. In this study, different pure bacteria from human feces were isolated and were used to investigate their conversion capability of astilbin. Ultra-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technique combined with Metabolynx(TM) software was used to analyze astilbin and its metabolites. The parent compound and two metabolites (quercetin and eriodictyol) were detected in the isolated bacterial samples compared with blank samples. Quercetin was present in Enterococcus sp. 8B, 8-2 and 9-2 samples. Eriodictyol was only identified in Enterococcus sp. 8B sample. The metabolic routes and metabolites of astilbin produced by the different intestinal bacteria are reported for the first time. This will be useful for the investigation of the pharmacokinetic study of astilbin in vivo and the role of different intestinal bacteria in the metabolism of natural compounds.

Amplified Electrochemiluminescent Aptasensor Using Mimicking Bi-enzyme Nanocomplexes As Signal Enhancement

In this work, a sandwich-type electrochemiluminescence (ECL) aptasensor for ultrasensitive detection of thrombin (TB) was designed based on mimicking bi-enzyme cascade catalysis to in situ generate coreactant of dissolved oxygen (O2) for signal amplification. We utilized hollow Au nanoparticles (HAuNPs) as carriers to immobilize glucose oxidase nanoparticles (GOxNPs) and Pt nanoparticles (PtNPs) by electrostatic adsorption. Then, the detection aptamer of thrombin (TBA 2) was immobilized on the PtNPs/GOxNPs/HAuNPs nanocomplexes. Finally, hemin was intercalated into the TBA 2 to obtain the hemin/G-quadruplex structure. The hemin/G-quadruplex was an interesting DNAzyme that commonly mimiced horseradish peroxidase (HRP). Herein, GOxNPs, hemin/G-quadruplex and PtNPs could form mimicking bi-enzyme cascade catalysis system to in situ generate dissolved O2 as coreactant in peroxydisulfate solution when the testing buffer contained proper amounts of glucose. This method had successfully overcome the disadvantage of difficulty to label the dissolved O2 and realized the ECL signal amplification. The experiment proved that the aptasensor had good linear relationship on low concentration of TB. The linear range was 1×10(-6)-10 nM, with a detection limit of 0.3 fM.

An Evidence-based Knowledgebase of Pulmonary Arterial Hypertension to Identify Genes and Pathways Relevant to Pathogenesis

Pulmonary arterial hypertension (PAH) is a major progressive form of pulmonary hypertension (PH) with more than 4800 patients in the United States. In the last two decades, many studies have identified numerous genes associated with this disease. However, there is no comprehensive research resource for PAH or other PH types that integrates various genetic studies and their related biological information. Thus, the number of associated genes, and their strength of evidence, is unclear. In this study, we tested the hypothesis that a web-based knowledgebase could be used to develop a biological map of highly interrelated, functionally important genes in PAH. We developed the pulmonary arterial hypertension knowledgebase (PAHKB, ), a comprehensive database with a user-friendly web interface. PAHKB extracts genetic data from all available sources, including those from association studies, genetic mutation, gene expression, animal model, supporting literature, various genomic annotations, gene networks, cellular and regulatory pathways, as well as microRNAs. Moreover, PAHKB provides online tools for data browsing and searching, data integration, pathway graphical presentation, and gene ranking. In the current release, PAHKB contains 341 human PH-related genes (293 protein coding and 48 non-coding genes) curated from over 1000 PubMed abstracts. Based on the top 39 ranked PAH-related genes in PAHKB, we constructed a core biological map. This core map was enriched with the TGF-beta signaling pathway, focal adhesion, cytokine-cytokine receptor interaction, and MAPK signaling. In addition, the reconstructed map elucidates several novel cancer signaling pathways, which may provide clues to support the application of anti-cancer therapeutics to PAH. In summary, we have developed a system for the identification of core PH-related genes and identified critical signaling pathways that may be relevant to PAH pathogenesis. This system can be easily applied to other pulmonary diseases.

A Stable Microporous Mixed-metal Metal-organic Framework with Highly Active Cu2+ Sites for Efficient Cross-dehydrogenative Coupling Reactions

Two metalloporphyrin octacarboxylates were used to link copper(II) nodes for the formation of two novel porous mixed-metal metal-organic frameworks (M'MOFs) containing nanopore cages (2.1 nm in diameter) or nanotubular channels (1.5 nm in diameter). The highly active Cu(2+) sites on the nanotubular surfaces of the stable porous M'MOF ZJU-22, stabilized by three-connected nets, lead to the superior catalytic activity for the cross-dehydrogenative coupling (CDC) reaction.

Ultra Performance Liquid Chromatography/quadrupole-time-of-flight Mass Spectrometry for Determination of Avicularin Metabolites Produced by a Human Intestinal Bacterium

Intestinal bacteria from human were screened to isolate the specific bacteria involved in the metabolism of avicularin. A Gram-positive anaerobic bacterium, strain 46, capable of metabolizing avicularin (quercetin-3-O-arabinoside) was isolated for the first time. Its 16S rRNA gene sequence showed 99% similarity with that of Bacillus. Then strain 46 was identified as a species of the genus Bacillus, and was named to be Bacillus sp. 46. Additionally, the metabolites were analyzed by ultra performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) technique combined with Metabolynx™ software. The structure of these metabolites were proposed and confirmed by comparing the UPLC retention time and MS/MS spectrum with that of authentic standards. Parent compound and six metabolites were detected in the isolated bacterial samples compared with blank samples. Avicularin (M1) was anaerobic metabolized to its aglycone quercetin (M2) and methoxylated avicularin (M3, M4), then quercetin was converted to quercetin glycosides: quercetin-3-O-rhamnoside (M5), quercetin-3-O-glucoside (M6) and quercetin-7-O-glucoside (M7) by Bacillus sp. 46. The metabolic pathway and metabolites of avicularin by the intestinal bacterium Bacillus sp. 46 were reported for the first time.

Porous Metal-organic Frameworks for Heterogeneous Biomimetic Catalysis

Metalloporphyrins are the active sites in monooxygenases that oxidize a variety of substrates efficiently and under mild conditions. Researchers have developed artificial metalloporphyrins, but these structures have had limited catalytic applications. Homogeneous artificial metalloporphyrins can undergo catalytic deactivation via suicidal self-oxidation, which lowers their catalytic activity and sustainability relative to their counterparts in Nature. Heme molecules in protein scaffolds can maintain high efficiency over numerous catalytic cycles. Therefore, we wondered if immobilizing metalloporphyrin moieties within porous metal-organic frameworks (MOFs) could stabilize these structures and facilitate the molecular recognition of substrates and produce highly efficient biomimetic catalysis. In this Account, we describe our research to develop multifunctional porphyrinic frameworks as highly efficient heterogeneous biomimetic catalysts. Our studies indicate that porous porphyrinic frameworks provide an excellent platform for mimicking the activity of biocatalysts and developing new heterogeneous catalysts that effect new chemical transformations under mild conditions. The porous structures and framework topologies of the porphyrinic frameworks depend on the configurations, coordination donors, and porphyrin metal ions of the metalloporphyrin moieties. To improve the activity of porous porphyrinic frameworks, we have developed a two-step synthesis that introduces the functional polyoxometalates (POMs) into POM-porphyrin hybrid materials. To tune the pore structures and the catalytic properties of porphyrinic frameworks, we have designed metalloporphyrin M-H8OCPP ligands with four m-benzenedicarboxylate moieties, and introduced the secondary auxiliary ligands. The porphyrin metal ions and the secondary functional moieties that are incorporated into porous metal-organic frameworks greatly influence the catalytic properties and activities of porphyrinic frameworks in different reactions, such as the oxidation of alkylbenzenes, olefins, and hexane and the photo-oxygenation of 1,5-dihydroxynaphthalene and sulfides. The porphyrin metal ions and the secondary auxiliary sites in the pores can work together synergistically to enhance the catalytic activities of porphyrinic frameworks. Compared with their homogeneous counterparts, the activities and stabilities of the heterogeneous porphyrinic frameworks are remarkable: the immobilization of metalloporphyrins onto the pore surfaces of MOFs not only prevents their suicidal self-oxidation but also allows them to activate inert substrate molecules, such as cyclohexane. Moreover, because the bulky molecules cannot easily access the active sites inside the pores of porphyrinic frameworks, these porous materials demonstrate interesting size-selective catalytic properties toward substrates.

Human Transporter Database: Comprehensive Knowledge and Discovery Tools in the Human Transporter Genes

Transporters are essential in homeostatic exchange of endogenous and exogenous substances at the systematic, organic, cellular, and subcellular levels. Gene mutations of transporters are often related to pharmacogenetics traits. Recent developments in high throughput technologies on genomics, transcriptomics and proteomics allow in depth studies of transporter genes in normal cellular processes and diverse disease conditions. The flood of high throughput data have resulted in urgent need for an updated knowledgebase with curated, organized, and annotated human transporters in an easily accessible way. Using a pipeline with the combination of automated keywords query, sequence similarity search and manual curation on transporters, we collected 1,555 human non-redundant transporter genes to develop the Human Transporter Database (HTD) (http://htd.cbi.pku.edu.cn). Based on the extensive annotations, global properties of the transporter genes were illustrated, such as expression patterns and polymorphisms in relationships with their ligands. We noted that the human transporters were enriched in many fundamental biological processes such as oxidative phosphorylation and cardiac muscle contraction, and significantly associated with Mendelian and complex diseases such as epilepsy and sudden infant death syndrome. Overall, HTD provides a well-organized interface to facilitate research communities to search detailed molecular and genetic information of transporters for development of personalized medicine.

Factors Associated with One Year Retention to Methadone Maintenance Treatment Program Among Patients with Heroin Dependence in China

The aim of this study was to evaluate the risk factors associated with dropout from Methadone Maintenance Treatment (MMT) clinics within a 1 year follow-up cohort study in China.

A Systems Biology Approach to Identify Intelligence Quotient Score-related Genomic Regions, and Pathways Relevant to Potential Therapeutic Treatments

Although the intelligence quotient (IQ) is the most popular intelligence test in the world, little is known about the underlying biological mechanisms that lead to the differences in human. To improve our understanding of cognitive processes and identify potential biomarkers, we conducted a comprehensive investigation of 158 IQ-related genes selected from the literature. A genomic distribution analysis demonstrated that IQ-related genes were enriched in seven regions of chromosome 7 and the X chromosome. In addition, these genes were enriched in target lists of seven transcription factors and sixteen microRNAs. Using a network-based approach, we further reconstructed an IQ-related pathway from known human pathway interaction data. Based on this reconstructed pathway, we incorporated enriched drugs and described the importance of dopamine and norepinephrine systems in IQ-related biological process. These findings not only reveal several testable genes and processes related to IQ scores, but also have potential therapeutic implications for IQ-related mental disorders.

Activation of ERK1/2 by NADPH Oxidase-originated Reactive Oxygen Species Mediates Uric Acid-induced Mesangial Cell Proliferation

Hyperuricemia is associated with kidney complications including glomerulosclerosis and mesangial cell (MC) proliferation by poorly understood mechanisms. The present study investigated the underlying mechanisms that mediate uric acid (UA)-induced MC proliferation. A rat MC line, HBZY-1, was treated with various concentrations of UA in the presence or absence of a specific extracellular-regulated protein kinase 1/2 (ERK1/2) inhibitor (U0126), apocynin. UA dose dependently stimulated MC proliferation as shown by increased DNA synthesis and number of cells in the S and G2 phases in parallel with the upregulation of cyclin A2 and cyclin D1. In addition, UA time dependently promoted MC proliferation and significantly increased phosphorylation of ERK1/2 but not c-Jun NH2-terminal kinase and p38 MAPK in MCs as assessed by immunoblotting. Inhibition of ERK1/2 signaling via U0126 markedly blocked UA-induced MC proliferation. More importantly, UA induced intracellular reactive oxygen species (ROS) production of MCs dose dependently, which was completely blocked by apocynin, a specific NADPH oxidase inhibitor. Toll-like receptor (TLR)2 and TLR4 signaling had no effect on NADPH-derived ROS and UA-induced MC proliferation. Interestingly, pretreatment with apocynin inhibited ERK1/2 activation, the upregulation of cyclin A2 and cyclin D1, and MC proliferation. In conclusion, UA-induced MC proliferation was mediated by NADPH/ROS/ERK1/2 signaling pathway. This novel finding not only reveals the mechanism of UA-induced MC cell proliferation but also provides some potential targets for future treatment of UA-related glomerular injury.

Ex Vivo Expanded Human Regulatory T Cells Delay Islet Allograft Rejection Via Inhibiting Islet-derived Monocyte Chemoattractant Protein-1 Production in CD34+ Stem Cells-reconstituted NOD-scid IL2rγnull Mice

Type 1 diabetes mellitus (T1DM) is an autoimmune disease caused by immune-mediated destruction of insulin-secreting β cells of the pancreas. Near complete dependence on exogenous insulin makes T1DM very difficult to control, with the result that patients are exposed to high blood glucose and risk of diabetic complications and/or intermittent low blood glucose that can cause unconsciousness, fits and even death. Allograft transplantation of pancreatic islets restores normoglycemia with a low risk of surgical complications. However, although successful immediately after transplantation, islets are progressively lost, with most of the patients requiring exogenous insulin within 2 years post-transplant. Therefore, there is an urgent requirement for the development of new strategies to prevent islet rejection. In this study, we explored the importance of human regulatory T cells in the control of islets allograft rejection. We developed a pre-clinical model of human islet transplantation by reconstituting NOD-scid IL2rγnull mice with cord blood-derived human CD34+ stem cells and demonstrated that although the engrafted human immune system mediated the rejection of human islets, their survival was significantly prolonged following adoptive transfer of ex vivo expanded human Tregs. Mechanistically, Tregs inhibited the infiltration of innate immune cells and CD4+ T cells into the graft by down-regulating the islet graft-derived monocyte chemoattractant protein-1. Our findings might contribute to the development of clinical strategies for Treg therapy to control human islet rejection. We also show for the first time that CD34+ cells-reconstituted NOD-scid IL2rγnull mouse model could be beneficial for investigating human innate immunity in vivo.

Analysis of the Metabolites of Isorhamnetin 3-O-glucoside Produced by Human Intestinal Flora in Vitro by Applying Ultraperformance Liquid Chromatography/quadrupole Time-of-flight Mass Spectrometry

Isorhamnetin 3-O-glucoside, which is widely contained in many vegetables and rice, is expected to be metabolized by intestinal microbiota after digestion, which brings about the profile of its pharmacological effect. However, little is known about the interactions between this active ingredient and the intestinal flora. In this study, the preculture bacteria and GAM (general anaerobic medium) broth with isorhamnetin 3-O-glucoside were mixed for 48 h of incubation. Ultraperformance liquid chromatography/quadrupole time-of-flight mass spectrometry was used for analysis of the metabolites of isorhamnetin 3-O-glucoside in the corresponding supernatants of fermentation. The parent and five metabolites were found and preliminarily identified on the basis of the chromatograms and characteristics of their protonated ions. Four main metabolic pathways, including deglycosylation, demethoxylation, dehydroxylation, and acetylation, were summarized to explain how the metabolites were converted. Acetylated isorhamnetin 3-O-glucoside and kaempferol 3-O-glucoside were detected only in the sample of Escherichia sp. 12, and quercetin existed only in the sample of Escherichia sp. 4. However, the majority of bacteria could metabolize isorhamnetin 3-O-glucoside to its aglycon isorhamnetin, and then isorhamnetin was degraded to kaempferol. The metabolic pathway and the metabolites of isorhamnetin 3-O-glucoside yielded by different isolated human intestinal bacteria were investigated for the first time. The results probably provided useful information for further in vivo metabolism and active mechanism research on isorhamnetin 3-O-glucoside.

Resource Optimized TTSH-URA for Multimedia Stream Authentication in Swallowable-capsule-based Wireless Body Sensor Networks

To ease the burdens on the hospitalization capacity, an emerging swallowable-capsule technology has evolved to serve as a remote gastrointestinal (GI) disease examination technique with the aid of the wireless body sensor network (WBSN). Secure multimedia transmission in such a swallowable-capsule-based WBSN faces critical challenges including energy efficiency and content quality guarantee. In this paper, we propose a joint resource allocation and stream authentication scheme to maintain the best possible video quality while ensuring security and energy efficiency in GI-WBSNs. The contribution of this research is twofold. First, we establish a unique signature-hash (S-H) diversity approach in the authentication domain to optimize video authentication robustness and the authentication bit rate overhead over a wireless channel. Based on the full exploration of S-H authentication diversity, we propose a new two-tier signature-hash (TTSH) stream authentication scheme to improve the video quality by reducing authentication dependence overhead while protecting its integrity. Second, we propose to combine this authentication scheme with a unique S-H oriented unequal resource allocation (URA) scheme to improve the energy-distortion-authentication performance of wireless video delivery in GI-WBSN. Our analysis and simulation results demonstrate that the proposed TTSH with URA scheme achieves considerable gain in both authenticated video quality and energy efficiency.

Authors' Response: Nuchal Cord and Cesarean Delivery in China

Biofeedback Combined with Cue-exposure As a Treatment for Heroin Addicts

The aim of this study was to test if cue-exposure therapy (CET) combined with biofeedback therapy (BT) could decrease craving and physiological reactivity to drug-related cues in heroin dependents. Forty-five participants were randomly assigned to usual rehabilitation with or without CET combined with BT. Craving was assessed by a 100-point visual analog scale (VAS). Skin conductance (SC) and muscle electromyography (MEG) were recorded using a biofeedback device. After 2 months of treatment, both the pre-cue exposure craving and the post-cue exposure craving, SC, and MEG were lower in the experimental group than in the control group. Compared to the control group, the experimental group had a greater decrease in craving, SC, and MEG from baseline after the treatment. The results suggest that CET combined with BT treatment is effective in reducing craving and physiology reactivity in heroin dependents and could be used as a component of heroin-dependence rehabilitation.

Comparative Metabolism of Radix Scutellariae Extract by Intestinal Bacteria from Normal and Type 2 Diabetic Mice in Vitro

Traditional Chinese medicine (TCM) has been used in clinical practice for several thousand years. TCM has played an indispensable role in the prevention and treatment of disease, especially the complicated and chronic ones. In TCMs, many ingredients which are known to have biological effects just pass through the gut, they do not get into the bloodstream. Study on interactions of these active ingredients with the intestinal bacteria is very helpful to unravel how TCM works. Radix scutellariae was widely used alone or in combination with other medicinal herbs to the treatment of type 2 diabetes mellitus for a long time in China even in Asia. Additionally, the incidence of type 2 diabetes is closely related to the changes of intestinal flora. In this paper, the metabolism of baicalin in Radix scutellariae extract by normal and type 2 diabetic mice intestinal bacteria were firstly investigated.

Cross-sectional Study of the Relationship of Peripheral Blood Cell Profiles with Severity of Infection by Adenovirus Type 55

The immunologic profiles of patients with human adenovirus serotype 55 (HAdV-55) infections were characterized in subjects diagnosed with silent infections (n = 30), minor infections (n = 27), severe infections (n = 34), and healthy controls (n = 30) during a recent outbreak among Chinese military trainees.

One-pot Synthesis of Highly Luminescent Carbon Quantum Dots and Their Nontoxic Ingestion by Zebrafish for in Vivo Imaging

Photoluminescent carbon and/or silicon-based nanodots have attracted ever increasing interest. Accordingly, a myriad of synthetic methodologies have been developed to fabricate them, which unfortunately, however, frequently involve relatively tedious steps, such as initial surface passivation and subsequent functionalization. Herein, we describe a green and sustainable synthetic strategy to combine these procedures into one step and to produce highly luminescent carbon quantum dots (CQDs), which can also be easily fabricated into flexible thin films with intense luminescence for future roll-to-roll manufacturing of optoelectronic devices. The as-synthesized CQDs exhibited enhanced cellular permeability and low or even noncytotoxicity for cellular applications, as corroborated by confocal fluorescence imaging of HeLa cells as well as cell viability measurements. Most strikingly, zebrafish were directly fed with CQDs for in vivo imaging, and mortality and morphologic analysis indicated ingestion of the CQDs posed no harm to the living organisms. Hence, the multifunctional CQDs potentially provide a rich pool of tools for optoelectronic and biomedical applications.

Different Hereditary Contribution of the CFH Gene Between Polypoidal Choroidal Vasculopathy and Age-related Macular Degeneration in Chinese Han People

To investigate whether 11 variants in complement factor H gene contributed differently in patients with neovascular age-related macular degeneration (nAMD) and polypoidal choroidal vasculopathy (PCV) of Chinese descent.

Chemical Composition and Biological Activities of Gerbera Anandria

Gerbera anandria (Compositae) was extracted with 75% ethanol and the residue was fractionated using light petroleum, chloroform and ethyl acetate. The constituents of the extracts were separated by column chromatography employing solvents of different polarity. Column chromatography of the light petroleum fraction resulted in the isolation of methyl hexadecanoate, while the chloroform fraction afforded xanthotoxin, 2-hydroxy-6-methylbenzoic acid, 7-hydroxy-1(3H)-isobenzofuranone, a mixture of β-sitosterol and stigmasterol, and 8-methoxysmyrindiol and the ethyl acetate fraction gave gerberinside, apigenin-7-O-β-d-glucopyranoside and quercetin. A new coumarin, 8-methoxysmyrindiol, was found. The chemical structures of the isolated compounds were established by MS and NMR (HSQC, HMBC). Free radical scavenging and cytotoxic activities of crude extracts and 8-methoxysmyrindiol were further investigated. The ethyl acetate phase exerted the strongest DPPH free radical scavenging activity in comparison to the other fractions. The coumarin 8-methoxysmyrindiol demonstrated cytotoxicity against multiple human cancer cell lines, with the highest potency in HepG2 cells.

Effect of Low Concentration Sodium Dodecyl Sulfate on the Electromigration of Palonosetron Hydrochloride Stereoisomers in Micellar Electrokinetic Chromatography

The effect of low concentrations of sodium dodecyl sulfate (SDS) on the separation of palonosetron hydrochloride (PALO) stereoisomers by micellar electrokinetic chromatography (MEKC) has been investigated. It was found that the addition of SDS prolongs the migration time and the migration order of four stereoisomers changes regularly with the SDS concentration. Good separations for all the four stereoisomers were achieved at appropriate SDS concentration. The effect of SDS on the electromigration (mobilities) of PALO stereoisomers has been studied, in order to explain its effect on the separation by MEKC. It was found that low concentrations of SDS added into the separation media forms negatively charged complexes with PALO stereoisomers and hence reverses their electromigration direction. Furthermore, the migration order between two enantiomeric pairs is also reversed because the enantiomeric pair with a bigger positive mobility than that of another pair turns to have a bigger negative mobility when bound with SDS. Based on these results, the effect of SDS on the MEKC separation of PALO stereoisomers was elucidated reasonably. The performance of the developed chiral MEKC method was validated by the analysis of a real sample.

The Electrical Response to Injury: Molecular Mechanisms and Wound Healing

Significance: Natural, endogenous electric fields (EFs) and currents arise spontaneously after wounding of many tissues, especially epithelia, and are necessary for normal healing. This wound electrical activity is a long-lasting and regulated response. Enhancing or inhibiting this electrical activity increases or decreases wound healing, respectively. Cells that are responsible for wound closure such as corneal epithelial cells or skin keratinocytes migrate directionally in EFs of physiological magnitude. However, the mechanisms of how the wound electrical response is initiated and regulated remain unclear. Recent Advances: Wound EFs and currents appear to arise by ion channel up-regulation and redistribution, which are perhaps triggered by an intracellular calcium wave or cell depolarization. We discuss the possibility of stimulation of wound healing via pharmacological enhancement of the wound electric signal by stimulation of ion pumping. Critical Issues: Chronic wounds are a major problem in the elderly and diabetic patient. Any strategy to stimulate wound healing in these patients is desirable. Applying electrical stimulation directly is problematic, but pharmacological enhancement of the wound signal may be a promising strategy. Future Directions: Understanding the molecular regulation of wound electric signals may reveal some fundamental mechanisms in wound healing. Manipulating fluxes of ions and electric currents at wounds might offer new approaches to achieve better wound healing and to heal chronic wounds.

The Influence of Drug Physical State on the Dissolution Enhancement of Solid Dispersions Prepared Via Hot-melt Extrusion: a Case Study Using Olanzapine

In this study, we examine the relationship between the physical structure and dissolution behavior of olanzapine (OLZ) prepared via hot-melt extrusion in three polymers [polyvinylpyrrolidone (PVP) K30, polyvinylpyrrolidone-co-vinyl acetate (PVPVA) 6:4, and Soluplus® (SLP)]. In particular, we examine whether full amorphicity is necessary to achieve a favorable dissolution profile. Drug–polymer miscibility was estimated using melting point depression and Hansen solubility parameters. Solid dispersions were characterized using differential scanning calorimetry, X-ray powder diffraction, and scanning electron microscopy. All the polymers were found to be miscible with OLZ in a decreasing order of PVP>PVPVA>SLP. At a lower extrusion temperature (160°C), PVP generated fully amorphous dispersions with OLZ, whereas the formulations with PVPVA and SLP contained 14%-16% crystalline OLZ. Increasing the extrusion temperature to 180°C allowed the preparation of fully amorphous systems with PVPVA and SLP. Despite these differences, the dissolution rates of these preparations were comparable, with PVP showing a lower release rate despite being fully amorphous. These findings suggested that, at least in the particular case of OLZ, the absence of crystalline material may not be critical to the dissolution performance. We suggest alternative key factors determining dissolution, particularly the dissolution behavior of the polymers themselves.

Finite Element Analysis of a Bone Healing Model: 1-year Follow-up After Internal Fixation Surgery for Femoral Fracture

Finite element analysis was used to compare preoperative and postoperative stress distribution of a bone healing model of femur fracture, to identify whether broken ends of fractured bone would break or not after fixation dislodgement one year after intramedullary nailing. Method s: Using fast, personalized imaging, bone healing models of femur fracture were constructed based on data from multi-slice spiral computed tomography using Mimics, Geomagic Studio, and Abaqus software packages. The intramedullary pin was removed by Boolean operations before fixation was dislodged. Loads were applied on each model to simulate a person standing on one leg. The von Mises stress distribution, maximum stress, and its location was observed. Results : According to 10 kinds of display groups based on material assignment, the nodes of maximum and minimum von Mises stress were the same before and after dislodgement, and all nodes of maximum von Mises stress were outside the fracture line. The maximum von Mises stress node was situated at the bottom quarter of the femur. The von Mises stress distribution was identical before and after surgery. Conclusion : Fast, personalized model establishment can simulate fixation dislodgement before operation, and personalized finite element analysis was performed to successfully predict whether nail dislodgement would disrupt femur fracture or not.

NHE3 Phosphorylation Via PKCη Marks the Polarity and Orientation of Directionally Migrating Cells

Endogenous electric fields (EF) may provide an overriding cue for directional cell migration during wound closure. Perceiving a constant direction requires active sodium-hydrogen exchanger (pNHE3) at the leading edge of HEK 293 cells but its activation mechanism is not yet fully understood. Because protein kinase C (PKC) is required in electrotaxis, we asked whether NHE3 is activated by PKC during wound healing. Using pharmacological (pseudosubstrate and edelfosine) inhibition, we showed that inhibition of PKCη isoform impairs directional cell migration in HEK 293 cells in the presence of a persistent directional cue (0.25-0.3 V/mm of EF for 2 h). Further, we found that pNHE3 forms complexes with both PKCη and ɣ-tubulin, suggesting that these molecules may regulate the microtubule-organizing center. In addition, cellular pNHE3 content was reduced significantly when PKCη was inhibited during directional cell migration. Taken together, these data suggest that PKCη-dependent phosphorylation of NHE3 and the formation of pNHE3/PKCη/ɣ-tubulin complexes at the leading edge of the cell are required for directional cell migration in an EF.

Catalase Mimic Property of Co3O4 Nanomaterials with Different Morphology and Its Application As a Calcium Sensor

The applications of inorganic nanomaterials as biomimetic catalysts are receiving much attention because of their high stability and low cost. In this work, Co3O4 nanomaterials including nanoplates, nanorods, and nanocubes were synthesized. The morphologies and compositions of the products were characterized by scanning electron microscopy, transmission electron microscopy, and X-ray diffraction. The catalytic properties of Co3O4 nanomaterials as catalase mimics were studied. The Co3O4 materials with different morphology exhibited different catalytic activities in the order of nanoplates > nanorods > nanocubes. The difference of the catalytic activities originated from their different abilities of electron transfer. Their catalytic activities increased significantly in the presence of calcium ion. On the basis of the stimulation by calcium ion, a biosensor was constructed by Co3O4 nanoplates for the determination of calcium ion. The biosensor had a linear relation to calcium concentrations and good measurement correlation between 0.1 and 1 mM with a detection limit of 4 μM (S/N = 3). It showed high selectivity against other metal ions and good reproducibility. The proposed method was successfully applied for the determination of calcium in a milk sample.

Angiotensin II Plasma Levels Are Linked to Disease Severity and Predict Fatal Outcomes in H7N9-infected Patients

A novel influenza A (H7N9) virus of avian origin emerged in eastern China in the spring of 2013. This virus causes severe disease in humans, including acute and often lethal respiratory failure. As of January 2014, 275 cases of H7N9-infected patients had been reported, highlighting the urgency of identifying biomarkers for predicting disease severity and fatal outcomes. Here, we show that plasma levels of angiotensin II, a major regulatory peptide of the renin-angiotensin system, are markedly elevated in H7N9 patients and are associated with disease progression. Moreover, the sustained high levels of angiotensin II in these patients are strongly correlated with mortality. The predictive value of angiotensin II is higher than that of C-reactive protein and some clinical parameters such as the PaO2/FiO2 ratio (partial pressure of arterial oxygen to the fraction of inspired oxygen). Our findings indicate that angiotensin II is a biomarker for lethality in flu infections.

Single Cell Wound Generates Electric Current Circuit and Cell Membrane Potential Variations That Requires Calcium Influx

Breaching of the cell membrane is one of the earliest and most common causes of cell injury, tissue damage, and disease. If the compromise in cell membrane is not repaired quickly, irreversible cell damage, cell death and defective organ functions will result. It is therefore fundamentally important to efficiently repair damage to the cell membrane. While the molecular aspects of single cell wound healing are starting to be deciphered, its bio-physical counterpart has been poorly investigated. Using Xenopus laevis oocytes as a model for single cell wound healing, we describe the temporal and spatial dynamics of the wound electric current circuitry and the temporal dynamics of cell membrane potential variation. In addition, we show the role of calcium influx in controlling electric current circuitry and cell membrane potential variations. (i) Upon wounding a single cell: an inward electric current appears at the wound center while an outward electric current is observed at its sides, illustrating the wound electric current circuitry; the cell membrane is depolarized; calcium flows into the cell. (ii) During cell membrane re-sealing: the wound center current density is maintained for a few minutes before decreasing; the cell membrane gradually re-polarizes; calcium flow into the cell drops. (iii) In conclusion, calcium influx is required for the formation and maintenance of the wound electric current circuitry, for cell membrane re-polarization and for wound healing.

Dysfunction of the PGC-1α-mitochondria Axis Confers Adriamycin-induced Podocyte Injury

Adriamycin (ADR)-induced nephropathy in animals is an experimental analog of human focal segmental glomerulosclerosis, which presents as severe podocyte injury and massive proteinuria and has a poorly understood mechanism. The present study was designed to test the hypothesis that the peroxisome proliferator-activated receptor-γ coactivator (PGC)-1α-mitochondria axis is involved in ADR-induced podocyte injury. Using MPC5 immortalized mouse podocytes, ADR dose dependently induced downregulation of nephrin and podocin, cell apoptosis, and mitochondrial dysfunction based on the increase in mitochondrial ROS production, decrease in mitochondrial DNA copy number, and reduction of mitochondrial membrane potential and ATP content. Moreover, ADR treatment also remarkably reduced the expression of PGC-1α, an important regulator of mitochondrial biogenesis and function, in podocytes. Strikingly, PGC-1α overexpression markedly attenuated mitochondrial dysfunction, the reduction of nephrin and podocin, and the apoptotic response in podocytes after ADR treatment. Moreover, downregulation of PGC-1α and mitochondria disruption in podocytes were also observed in rat kidneys with ADR administration, suggesting that the PGC-1α-mitochondria axis is relevant to in vivo ADR-induced podocyte damage. Taken together, these novel findings suggest that dysfunction of the PGC-1α-mitochondria axis is highly involved in ADR-induced podocyte injury. Targeting PGC-1α may be a novel strategy for the treatment of ADR nephropathy and human focal segmental glomerulosclerosis.

[Effect of Weifuchun on Inhibiting Inflammation of Helicobacter Pylori-infected GES-1 Cells and NF-kappaB Signaling Pathway]

To study the effect of Weifuchun on inflammation of Helicobacter pylori (Hp)-infected gastric epithelial cells (GES-1) and its correlation with NF-kappaB signaling pathway.

Davida Teller Award Lecture 2013: the Importance of Prediction and Anticipation in the Control of Smooth Pursuit Eye Movements

The ability of smooth pursuit eye movements to anticipate the future motion of targets has been known since the pioneering work of Dodge, Travis, and Fox (1930) and Westheimer (1954). This article reviews aspects of anticipatory smooth eye movements, focusing on the roles of the different internal or external cues that initiate anticipatory pursuit.We present new results showing that the anticipatory smooth eye movements evoked by different cues differ substantially, even when the cues are equivalent in the information conveyed about the direction of future target motion. Cues that convey an easily interpretable visualization of the motion path produce faster anticipatory smooth eye movements than the other cues tested, including symbols associated arbitrarily with the path, and the same target motion tested repeatedly over a block of trials. The differences among the cues may be understood within a common predictive framework in which the cues differ in the level of subjective certainty they provide about the future path. Pursuit may be driven by a combined signal in which immediate sensory motion, and the predictions about future motion generated by sets of cues, are weighted according to their respective levels of certainty. Anticipatory smooth eye movements, an overt indicator of expectations and predictions, may not be operating in isolation, but may be part of a global process in which the brain analyzes available cues, formulates predictions, and uses them to control perceptual, motor, and cognitive processes.

The Levels of Ki-67 Positive Are Positively Associated with Lymph Node Metastasis in Invasive Ductal Breast Cancer

Breast cancer is the most common cancer in women worldwide. In this study, we evaluate the potential risk factors for lymph node metastasis in invasive breast cancer patients with axillary dissection. 147 patients were included into this prospective study. The prognostic biomarkers including Ki-67, human epidermal growth factor receptor 2 (HER-2), hormone receptor status, p53, and lymph node involvement were determined by immunohistochemistry. The association between lymph node metastasis and these biomarkers was analyzed. Lymph node metastasis was found in 62 patients out of 147 patients. The high levels of Ki-67 positive (greater than 20 %) were positively correlated with a higher incidence of lymph node metastasis, including the numbers of lymph nodes that contain tumor cells and the lymph node metastatic rate. The high rate of positive lymphovascular invasion (LVI) is associated with lymph node metastasis. However, the levels of Ki-67 positive were not correlated with the positive rate of LVI. There was also no association between lymph node metastasis and other prognostic biomarkers, such as HER-2, estrogen receptor, progesterone receptor, and p53. In addition, apart from p53, the levels of Ki-67 positive were correlated with other prognostic biomarkers. Our data suggest that Ki-67 positivity has value as a prognostic and predictive biomarker in breast cancer and may be a valuable proliferation marker in routine diagnosis of breast cancer.

Identification of the Metabolites of Myricitrin Produced by Human Intestinal Bacteria in Vitro Using Ultra-performance Liquid Chromatography/quadrupole Time-of-flight Mass Spectrometry

To investigate the metabolic routes and metabolites of myricitrin, an important active ingredient of traditional herbal medicine, yielded by the isolated human intestinal bacteria, which have not been reported previously.

A Luminescent Mixed-lanthanide-organic Framework Sensor for Decoding Different Volatile Organic Molecules

A flexible tripodal polyaromatic acid (4,4',4″-(((2,4,6-trimethylbenzene-1,3,5-triyl)-tris(methylene))-tris(oxy))tribenzoic acid, H3TCM) was used to adapt the coordination sites of lanthanide ions for the construction of microporous lanthanide-organic frameworks (LOFs) [LnTCM(H2O)2]·3DMF·H2O (Ln-TCM; Ln = La, Eu, and/or Tb). In these LOFs, the emission band of TCM matches well with the excitation energy of lanthanide ions (Eu(3+) and Tb(3+)) which results in high-efficient resonance energy transfer from TCM to lanthanide ions. Moreover, the mixed EuxTb1-x-TCM has tunable pores to adapt different induced-fit-type host-guest interactions which can modulate both the energy transfer efficiency from TCM to Ln(3+) ions and the energy allocation between Eu(3+) and Tb(3+) ions in the luminescence spectra. We demonstrate that the Eu(x)Tb(1-x)-TCM sensor has the capability of decoding different volatile organic molecules (VOMs) with a clearly differentiable and unique emission intensity ratio of (5)D0 → (7)F2 (Eu(3+), 614 nm) to (5)D4 → (7)F5 (Tb(3+), 545 nm) transitions for every different VOM. Compared with the traditional absolute emission intensity method, such a self-referencing emission intensity strategy has generated self-calibrating, highly differentiable, and very stable luminescent signals for decoding different VOMs from the unique Eu(x)Tb(1-x)-TCM platform, which has great potential for practical applications.

Associations of Cigarette Smoking and Alcohol Consumption with Metabolic Syndrome in a Male Chinese Population: a Cross-sectional Study

Whether cigarette smoking and alcohol consumption are associated with the risk of metabolic syndrome (MetS) remains controversial. This study investigated the associations of cigarette smoking and alcohol consumption with MetS in a male population in China.

Preparation of Paraoxonase-1 Liposomes and Studies on Their in Vivo Pharmacokinetics in Rats

A liposome formulation of the enzyme paraoxonase-1 (PON1) was prepared for purposes of prolonging and maintaining its activity in vivo. Following purification of PON1 from rabbit serum, liposomes containing PON1 (L-PON1) were prepared using a film-dispersion method with a soybean phospholipid-cholesterol mixture (5 : 1, w/w). The pharmacokinetic behaviour of conventional injectable PON1 and L-PON1 was compared following a single intravenous injection in rats. The enzyme activity of PON1 and its pharmacokinetic parameters were calculated based on a two-compartment model following conventional injection. The level of PON1 encapsulation in L-PON1 was 86.20 ± 3.12%. The particle size distribution of L-PON1 was a narrow unimodal form, with an average diameter of 126 nm. The results suggest that compared with conventional injectable PON1, L-PON1 has an improved half-life and enhanced enzyme activity in rats. In conclusion, PON1 can be encapsulated into a lipid bilayer for enhanced stability.

Polarizing Intestinal Epithelial Cells Electrically Through Ror2

The apicobasal polarity of enterocytes determines where the brush border membrane (apical membrane) will form, but how this apical membrane faces the lumen is not well understood. The electrical signal across the epithelium could serve as a coordinating cue, orienting and polarizing enterocytes. Here, we show that applying a physiological electric field to intestinal epithelial cells, to mimic the natural electric field created by the transepithelial potential difference, polarized phosphorylation of the actin-binding protein ezrin, increased expression of intestinal alkaline phosphatase (ALPI, a differentiation marker) and remodeled the actin cytoskeleton selectively on the cathode side. In addition, an applied electric field also activated ERK1/2 and LKB1 (also known as STK11), key molecules in apical membrane formation. Disruption of the tyrosine protein kinase transmembrane receptor Ror2 suppressed activation of ERK1/2 and LKB1 significantly, and subsequently inhibited apical membrane formation in enterocytes. Our findings indicate that the endogenous electric field created by the transepithelial potential difference might act as an essential coordinating signal for apical membrane formation at a tissue level, through activation of LKB1 mediated by Ror2-ERK signaling.

Effects of a Gemcitabine Plus Platinum Regimen Combined with a Dendritic Cell-cytokine Induced Killer Immunotherapy on Recurrence and Survival Rate of Non-small Cell Lung Cancer Patients

The aim of the present study was to investigate the effects of a gemcitabine plus platinum (GP) regimen combined with dendritic cell-cytokine induced killer (DC-CIK) immunotherapy on the recurrence and survival rate in patients with non-small cell lung cancer (NSCLC). Patients (n=157) with stage III NSCLC that had received surgery were randomly divided into a control group and an observation group. The control group was administered with a GP regimen and the observation group received GP chemotherapy that was based on DC-CIK cell immunotherapy in addition to SC-CIK cell immunotherapy here. The two groups were followed up for 36 months and their postoperative cellular immune function, disease-free survival time, cumulative recurrence rate and cumulative survival rate was analyzed. The percentages of CD3(+)CD4(+) T lymphocytes, natural killer cells and the CD4/CD8 ratio were identified to be significantly increased following treatment compared with those observed prior to treatment in the control and observation groups; conversely, the CD3(+)CD8(+) T lymphocyte percentage decreased significantly (P<0.05). Furthermore, the results of the patients in the observation group were significantly better compared with the control group based on these indicators (P<0.05). The median disease-free survival time of patients in the observation group (28 months) was identified to be significantly longer than that of the control group (22 months; P<0.05), the three-year cumulative recurrence rate in the observation group (47.37%) was significantly lower than that of the control group (76.92%; P<0.05) and the three-year cumulative survival rate of the patients in the observation group (58.23%) was significantly higher than that of the control group patients (37.14%; P<0.05). In conclusion, the GP regimen combined with DC-CIK immunotherapy significantly improved the immune cell function in the postoperative NSCLC patients, in addition to reducing postoperative tumor recurrence and prolonging the survival time of patients with NSCLC.

Valproic Acid Protects Septic Mice from Renal Injury by Reducing the Inflammatory Response

Valproic acid (VPA), a histone deacetylase inhibitor, has extensive activities against inflammation, oxidation, and malignancy. This study was designed to investigate the protective effect of VPA on the systemic inflammatory response and renal injury in septic mice.

CB2 Receptor Activation Ameliorates the Proinflammatory Activity in Acute Lung Injury Induced by Paraquat

Paraquat, a widely used herbicide, is well known to exhibit oxidative stress and lung injury. In the present study, we investigated the possible underlying mechanisms of cannabinoid receptor-2 (CB2) activation to ameliorate the proinflammatory activity induced by PQ in rats. JWH133, a CB2 agonist, was administered by intraperitoneal injection 1 h prior to PQ exposure. After PQ exposure for 4, 8, 24, and 72 h, the bronchoalveolar lavage fluid was collected to determine levels of TNF-α and IL-1β, and the arterial blood samples were collected for detection of PaO2 level. At 72 h after PQ exposure, lung tissues were collected to determine the lung wet-to-dry weight ratios, myeloperoxidase activity, lung histopathology, the protein expression level of CB2, MAPKs (ERK1/2, p38MAPK, and JNK1/2), and NF-κBp65. After rats were pretreated with JWH133, PQ-induced lung edema and lung histopathological changes were significantly attenuated. PQ-induced TNF-α and IL-1β secretion in BALF, increases of PaO2 in arterial blood, and MPO levels in the lung tissue were significantly reduced. JWH133 could efficiently activate CB2, while inhibiting MAPKs and NF-κB activation. The results suggested that activating CB2 receptor exerted protective activity against PQ-induced ALI, and it potentially contributed to the suppression of the activation of MAPKs and NF-κB pathways.

[Involvement of Toll-like Receptor 4 in Apoptosis of Hippocampal Neurons Through Akt/FoxO3a/Bim Signaling Pathways]

The present study was to investigate whether Toll-like receptor 4 (TLR4)-mediated Akt/FoxO3a/Bim signaling pathway participated in lipopolysaccharide (LPS)-induced apoptosis in hippocampal neurons. The primarily cultured rat hippocampal neurons were treated with LPS, TLR4 antibody+LPS, and LY294002+LPS, respectively. Cell vitality was assayed by CCK-8. Expressions of p-Akt, Akt, p-FoxO3a, FoxO3a, Bim and active-Caspase-3 of each group were detected by Western blot analysis; the mRNA expression of Bim was detected by real-time quantitative PCR; FoxO3a nuclear translocation was detected by fluorescence microscope. The rate of cell apoptosis was assayed by flow cytometry. The results showed that cell vitality of hippocampal neurons decreased after being treated with LPS in a time-dependent way. Compared with the control group, the expressions of p-Akt and p-FoxO3a decreased significantly, FoxO3a translocated into the nucleus, meanwhile, the expression of Bim and active-Caspase-3, and the apoptotic ratio of hippocampal neurons increased in LPS treated neurons. Pretreatment with TLR4 antibody significantly blocked, while PI3K antagonist LY294002 further strengthened these changes induced by LPS. In conclusion, the present study suggests that Akt/FoxO3a/Bim signaling pathways mediated by TLR4 participate in the apoptotic processes of primarily cultured hippocampal neurons treated with LPS, and the activation of TLR4 causes neuronal apoptosis.

Anti-vascular Endothelial Growth Factor Acts on Retinal Microglia/macrophage Activation in a Rat Model of Ocular Inflammation

To evaluate whether anti-vascular endothelial growth factor (VEGF) neutralizing antibodies injected in the vitreous of rat eyes influence retinal microglia and macrophage activation. To dissociate the effect of anti-VEGF on microglia and macrophages subsequent to its antiangiogenic effect, we chose a model of acute intraocular inflammation.

Expression of SATB1 Promotes the Growth and Metastasis of Colorectal Cancer

Special AT-rich sequence-binding protein-1 (SATB1) has been identified as a genome organizer that reprograms chromatin organization and transcription profiles. SATB1 promotes tumor growth and metastasis in breast cancer and is associated with poor prognosis in several cancer types. The association between SATB1 and colorectal cancer (CRC) has not been studied intensively. Therefore, this study aimed to investigate the effect of SATB1 on CRC growth and metastasis in vitro and in vivo and its correlation with overall survival and clinicopathological factors in CRC patients. Stable SATB1 knockdown and SATB1-overexpressing cell lines were established. SATB1 knockdown decreased cell growth, colony formation, migration, and invasion and increased apoptosis in CRC cells in vitro (p<0.05), whereas SATB1 overexpression had the opposite effect. SATB1 overexpression increased tumor growth and metastasis to lung and liver in vivo by using xenograft animal models (p<0.05). Thus, SATB1 promoted an aggressive CRC phenotype in vitro and in vivo. Immunohistochemical analysis of 560 CRC specimens showed that SATB1 expression was significantly higher in CRC tissues than in matched non-tumor mucosa (p<0.001). In addition, SATB1 expression was significantly higher in patients with poorly differentiated tumors, higher invasion depth, distant metastasis, and advanced TNM stage. SATB1-positive patients had a poorer prognosis than SATB1-negative patients, and SATB1 was identified as an independent prognostic factor for CRC (p = 0.009). Strikingly, we also evaluated SATB2 expression in CRC and found that SATB2 was more abundantly expressed in non-cancerous mucosa compared to colorectal cancer tissues (p<0.001). However, SATB2 expression had no influence on prognosis of CRC patients (p = 0.836). SATB1 expression was significantly associated with shorter survival time either in SATB2-positive patients or in SATB2-negative patients (p<0.001). In conclusion, our findings indicated an important role for SATB1 in CRC tumorigenesis and metastasis. Therefore, SATB1 may represent an important prognostic biomarker and therapeutic target for CRC.

MiR-20a Contributes to Endometriosis by Regulating NTN4 Expression

Endometriosis is a chronic disease that affects roughly 5-15 % of women of reproductive age. The pathophysiology of the disease occurrence and progression is unclear. MicroRNAs (miRNAs) are short, non-coding RNAs that have important regulatory function. It has been postulated that abnormal expression of miRNA is associated with ovarian endometriosis. Forty patients with ovarian endometriosis and 20 controls with benign ovarian tumor were included to examine the expression level of miR-20a. Quantitative real-time PCR (qPCR) was performed to detect the expression level of miR-20a. The target genes and pathways involved in aberrantly expressed miR-20a were identified by computational algorithms. Furthermore, selected target genes expression level were analyzed by qPCR. Significantly increased miR-20a expression level was observed in patients with ovarian endometriosis as compared with controls. Further stratified analysis showed that the increased expression level of miR-20a was only associated with advanced endometriosis (stage III-IV), but not mild endometriosis (stage I-II). The cell cycle was identified to be one of the most relevant pathways in the pathogenesis of endometriosis conducted by miR-20a. The expression level of target gene NTN4 (netrin-4) was significantly decreased in patients with ovarian endometriosis. The results of this study suggest that increased expression of miR-20a may play an important role in the pathogenesis of ovarian endometriosis by suppressing NTN4.

Comparative Metabolites in Plasma and Urine of Normal and Type 2 Diabetic Rats After Oral Administration of the Traditional Chinese Scutellaria-coptis Herb Couple by Ultra Performance Liquid Chromatography-tandem Mass Spectrometry

Scutellaria-coptis herb couple is widely used traditional Chinese medicine (TCM) in treating type 2 diabetes; however, the in vivo integrated metabolism of its main bioactive components in type 2 diabetic rats remains unknown. In this paper, ultra-performance liquid chromatography (UPLC) coupled to quadrupole time-of-flight (Q-TOF) and the MetaboLynx™ software combined with mass defect filtering (MDF) together provided unique high throughput capabilities for drug metabolism study with excellent MS mass accuracy and enhanced MS(E) data acquisition. This rapid automated analysis method was successfully applied for screening and identification of the absorbed and metabolized constituents after oral administration of scutellaria-coptis extract to rats. The results showed that a total of 14 metabolites of two parent compounds were detected and tentatively identified in vivo based on the characteristics of their protonated ions. Main parent components of scutellaria-coptis extract such as baicalin and berberine were absorbed into the blood circulation of the rats. Differences of metabolite classes were not observed between normal and type 2 diabetic rat plasma and urine samples. However, the concentrations of baicalin and methylated berberine in type 2 diabetic rat plasma were much higher than those in normal sample. While, the concentrations of these two compounds in type 2 diabetic rat urine were remarkably lower than those in normal sample. This helped maintain a high blood drug concentration which might be beneficial for the treatment of type 2 diabetes. Additionally, the developed method was simple and reliable, revealing that it could be used to rapid screen and propose the structures of active components responsible for pharmacological effects of scutellaria-coptis and to better clarify its action mechanism. This work suggests that the integrative metabolism approach makes a useful template for drug metabolism research of TCMs.

Reliability and Validity of the Chinese Version of the Patient Health Questionnaire (PHQ-9) in the General Population

Depression is one of the most common mental illnesses. The reliability and the validity of the Patient Health Questionnaire (PHQ)-9, a depression screening tool, have not been examined in the general population in China. Thus, this study evaluated the reliability and the validity of the Chinese version of the PHQ-9 in detecting major depression in residents of a Chinese community.

Reproducible Combinatorial Regulatory Networks Elucidate Novel Oncogenic MicroRNAs in Non-small Cell Lung Cancer

While previous studies reported aberrant expression of microRNAs (miRNAs) in non-small cell lung cancer (NSCLC), little is known about which miRNAs play central roles in NSCLC's pathogenesis and its regulatory mechanisms. To address this issue, we presented a robust computational framework that integrated matched miRNA and mRNA expression profiles in NSCLC using feed-forward loops. The network consists of miRNAs, transcription factors (TFs), and their common predicted target genes. To discern the biological meaning of their associations, we introduced the direction of regulation. A network edge validation strategy using three independent NSCLC expression profiling data sets pinpointed reproducible biological regulations. Reproducible regulation, which may reflect the true molecular interaction, has not been applied to miRNA-TF co-regulatory network analyses in cancer or other diseases yet. We revealed eight hub miRNAs that connected to a higher proportion of targets validated by independent data sets. Network analyses showed that these miRNAs might have strong oncogenic characteristics. Furthermore, we identified a novel miRNA-TF co-regulatory module that potentially suppresses the tumor suppressor activity of the TGF-β pathway by targeting a core pathway molecule (TGFBR2). Follow-up experiments showed two miRNAs (miR-9-5p and miR-130b-3p) in this module had increased expression while their target gene TGFBR2 had decreased expression in a cohort of human NSCLC. Moreover, we demonstrated these two miRNAs directly bind to the 3' untranslated region of TGFBR2. This study enhanced our understanding of miRNA-TF co-regulatory mechanisms in NSCLC. The combined bioinformatics and validation approach we described can be applied to study other types of diseases.

Health Management in China

Trends in Abdominal Obesity Among U.S. Children and Adolescents

Previous studies showed that prevalence of abdominal obesity among US children and adolescents increased significantly between 1988-1994 and 2003-2004. However, little is known about recent time trends in abdominal obesity since 2003-2004.This study was to provide recent updated national estimates of childhood abdominal obesity and examine the trends in childhood abdominal obesity from 2003 to 2012.

NLRP3 Inflammasome Mediates Albumin-induced Renal Tubular Injury Through Impaired Mitochondrial Function

Proteinuria serves as a direct causative factor of renal tubular cell injury and is highly associated with the progression of chronic kidney disease via uncertain mechanisms. Recently, evidence demonstrated that both NLRP3 inflammasome and mitochondria are involved in the chronic kidney disease progression. The present study was undertaken to examine the role of NLRP3 inflammasome/mitochondria axis in albumin-induced renal tubular injury. In patients with proteinuria, NLRP3 was significantly up-regulated in tubular epithelial cells and was positively correlated with the severity of proteinuria. In agreement with these results, albumin remarkably activated NLRP3 inflammasome in both in vitro renal tubular cells and in vivo kidneys in parallel with significant epithelial cell phenotypic alteration and cell apoptosis. Genetic disruption of NLRP3 inflammasome remarkably attenuated albumin-induced cell apoptosis and phenotypic changes under both in vitro and in vivo conditions. In addition, albumin treatment resulted in a significant mitochondrial abnormality as evidenced by the impaired function and morphology, which was markedly reversed by invalidation of NLRP3/caspase-1 signaling pathway. Interestingly, protection of mitochondria function by Mn(III)tetrakis (4-benzoic acid) porphyrin (MnTBAP) or cyclosporin A (CsA) robustly attenuated albumin-induced injury in mouse proximal tubular cells. Collectively, these findings demonstrated a pathogenic role of NLRP3 inflammasome/caspase-1/mitochondria axis in mediating albumin-induced renal tubular injury. The discovery of this novel axis provides some potential targets for the treatment of proteinuria-associated renal injury.

Hypoglycemic Effect of Gynostemma Pentaphyllum Saponins by Enhancing the Nrf2 Signaling Pathway in STZ-inducing Diabetic Rats

Gynostemma pentaphyllum (GP) is a natural plant resources for diabetes therapy, however, there is little research on the mechanisms of GP. The present study was undertaken to characterize if G. pentaphyllum saponins (GPs) is the principal active compound of GP responsible for anti-diabetes, and to examine the relativity between blood glucose modulate and antioxidation. The GPs-treated streptozotocin diabetic rats had a more effective hypoglycemic status than those of diabetic control rats, which also ameliorate dyslipidemia. GPs has increased SOD and GSH-px activities, and the spleen and thymus indexes in diabetic rats. The insulin levels in the GPs-treated groups were significantly higher than diabetic control group. Our finding provides a new insight into the application of GPs for the treatment of oxidative stress related diseases.

[Influence of Different Skin Reactions of Acupoint-application on Clinical Outcomes in the Prevention and Treatment of Bronchial Asthma]

To observe the influence of different skin reactions of acupoint-application in summer on outcomes of prevention and treatment of bronchial asthma.

Use Trajectories of Amphetamine-type Stimulants (ATS) in Shanghai, China

Although amphetamine-type stimulant (ATS) use is an important issue that has caused growing concerns in China as well as other countries, the knowledge of long-term patterns of ATS use in China is limited. This study explored long-term patterns of ATS use in Shanghai, China, and compared the differences by ATS use trajectory groups, seeking to identify risk factors that have implications for the development of targeted intervention programs.

Polyethyleneglycol Diacrylate Microspheres: a Novel Carrier for Laccase Immobilisation

Abstract Laccase was immobilised on polyethyleneglycol diacrylate (PEGDA) microspheres. The optimal preparation conditions of PEGDA microspheres were as follows: 3.0% (w/v) 2,2-azobisisobutyro-nitrite (AIBN), 4.0-5.0% (w/v) polyvinylpyrrolidone (PVP), 5.0-8.0% (w/v) glucose and 4.0% (w/v) PEGDA in glucose solution. The volume ratio of PEGDA solution, glucose/PVP solution and AIBN solution was 25: 100: 1. Microspheres obtained exhibited good characteristics with small sizes (1-4 µm). The immobilised laccase showed a higher stability in a wide pH range. Thermal stability and storage stability of immobilised laccase were enhanced. The activity of immobilised laccase was 45.0% after six cycles uses. Only 62.7% of the activity remained for free laccase while there was a 60.4% increased for immobilised laccase with storage at 4 °C for 25 d. The Km value of laccase increased from 21.9 to 114.0 µmol/l after immobilisation.

Retinal Ischemia/reperfusion Injury is Mediated by Toll-like Receptor 4 Activation of NLRP3 Inflammasomes

Retinal ischemia/reperfusion (IR) is common in eye disorders. Pattern-recognition receptors (PRRs) are reported to initiate sterile inflammatory response. The role of PRRs in retinal IR injury is currently unknown. Thus, we investigated the expression and function of membrane and cytoplasmic PRRs during retinal IR.

Determination of the Enantiomeric and Diastereomeric Impurities of RS-glycopyrrolate by Capillary Electrophoresis Using Sulfated-β-cyclodextrin As Chiral Selectors

A practical chiral CE method, using sulfated-β-CD as chiral selector, was developed for the enantioseparation of glycopyrrolate containing two chiral centers. Several parameters affecting the separation were studied, including the nature and concentration of the chiral selectors, BGE pH, buffer type and concentration, separation voltage, and temperature. The separation was carried out in an uncoated fused-silica capillary of (effective length 40 cm) × 50 μm id with a separation voltage of 20 kV using 30 mM sodium phosphate buffer (pH 7.0, adjusted with 1 M sodium hydroxide) containing 2.0% w/v sulfated-β-CD at 25°C. Finally, the method for determining the enantiomeric impurities of RS-glycopyrrolate was proposed. The method was further validated with respect to its specificity, linearity range, accuracy and precision, LODs, and quantification in the expected range of occurrence for the isomeric impurities (0.1%).

Genotoxicity of Tri- and Hexavalent Chromium Compounds in Vivo and Their Modes of Action on DNA Damage in Vitro

Chromium occurs mostly in tri- and hexavalent states in the environment. Hexavalent chromium [Cr(VI)] compounds are extensively used in diverse industries, and trivalent chromium [Cr(III)] salts are used as micronutrients and dietary supplements. In the present work, we report that they both induce genetic mutations in yeast cells. They both also cause DNA damage in both yeast and Jurkat cells and the effect of Cr(III) is greater than that of Cr(VI). We further show that Cr(III) and Cr(VI) cause DNA damage through different mechanisms. Cr(VI) intercalates DNA and Cr(III) interferes base pair stacking. Based on our results, we conclude that Cr(III) can directly cause genotoxicity in vivo.

The Metabolism and Growth of Web Forums

We view web forums as virtual living organisms feeding on user's clicks and investigate how they grow at the expense of clickstreams. We find that PV(t) (the number of page views in a given time period) and UV(t) (the number of unique visitors in the time period) of the studied forums satisfy the law of the allometric growth, i.e., PV(t) ~ UV(t)((θ). We construct clickstream networks and explain the observed temporal dynamics of networks by the interactions between nodes. We describe the transportation of clickstreams using the function D(i) ~ T(i)(γ), in which T(i) is the total amount of clickstreams passing through node i and D(i) is the amount of the clickstreams dissipated from i to the environment. It turns out that γ, an indicator for the efficiency of network dissipation, not only negatively correlates with θ, but also sets the bounds for θ. In particular, 1/γ > θ when 0 < γ < 1 and 1/γ < θ when γ > 1. Our findings have practical consequences. For example, θ can be used as a measure of the "stickiness" of forums, which quantifies the stable ability of forums to remain users "lock-in" on the forum. Meanwhile, the correlation between γ and θ provides a method to predict the long-term "stickiness" of forums from the clickstream data in a short time period. Finally, we discuss a random walk model that replicates both of the allometric growth PV(t) ~ UV(t)(θ) and the dissipation function D(i) ~ T(γ)(i).

Placental Growth Factor Expression is Reversed by Antivascular Endothelial Growth Factor Therapy Under Hypoxic Conditions

Clinical trials have revealed that the antivascular endothelial growth factor (VEGF) therapies are effective in retinopathy of prematurity (ROP). But the low level of VEGF was necessary as a survival signal in healthy conditions, and endogenous placental growth factor (PIGF) is redundant for development. The purpose of this study was to elucidate the PIGF expression under hypoxia as well as the influence of anti-VEGF therapy on PIGF.

Comparison of Chinese and International Psychiatrists' Views on Classification of Mental Disorders

This study aimed to explore the views and attitudes of Chinese psychiatrists on mental disorders classification, and to compare their similarities and differences with those of the international mental health professionals.

Nut Consumption Decreases Risk of Some Diseases

Prevalence Analysis of Different Human Bocavirus Genotypes in Pediatric Patients Revealed Intra-genotype Recombination

Human bocavirus (HBoV) genotypes 1-4 have been detected worldwide in respiratory samples and stool samples, and are increasingly associated with respiratory and intestinal infections of previously unknown etiology in young children. Several studies revealed evidence of extensive recombination among HBoV genotypes at the NP1 and VP1 gene boundary region. This study explored the prevalence of HBoV genotypes in pediatric patients in Beijing, and studied their phylogeny.

Role of Biphasic Changes in Splenic Dendritic Cell Activity in a Mouse Model of Multiple Organ Dysfunction Syndrome

To analyze the changes in splenic dendritic cell (DC) activity and serum cytokine levels during the progression of multiple organ dysfunction syndrome (MODS). A C57BL/6 mouse model of MODS was established by intraperitoneal injection of zymosan. Immunohistochemistry and flow cytometry were used to detect expression of I-A(b) (MHC-II molecules of mice) as well as co-stimulatory and co-inhibitory molecules in spleen and DC surface. The levels of various cytokines in serum and spleen tissue were analyzed 6 h, 12 h, 24 h, 48 h, 5 d and 12 d after injury. Death occurred at 24-48 h and 10-12 d after injury. The expression of I-A(b) and CD86 in spleen tissue and on DCs increased 6-12 h after injury, followed by gradual reduction and at 12 d. The inhibitory molecule, PD-L1, was expressed on normal DCs, but expression of PD-1 was undetectable. PD-L1 and PD-1 expression increased and remained high at 5 d and 12 d after injury. In addition, TNF and IL-1 levels increased 6-12 h after injury; HMGB1 and IL-10 levels increased 24 h and 5 d after injury, respectively. In contrast, IL-2 and IL-12 decreased with disease progression. At 12 d after injury, proinflammatory and anti-inflammatory cytokine levels remained high, while IL-2 and IL-12 were significantly reduced. IL-10 and IL-12 changes in spleen were consistent with those in serum. MODS progression was characterized by changes in splenic DC activity as well as altered serum pro-inflammatory and anti-inflammatory cytokine levels, suggesting early immune activation and predominant immune tolerance at the late stage.

Circulating MicroRNA 483-5p As a Novel Biomarker for Diagnosis Survival Prediction in Multiple Myeloma

Dysregulation of miRNA expression plays an important role in cancer development, and circulating miRNAs are biomarkers of several cancers. We explored whether the miRNAs in plasma could be useful clinical biomarkers for multiple myeloma. miRNA microarray was conducted to identify elevation of four miRNAs and reduced levels of eight miRNAs in the plasma of nine multiple myeloma patients and seven healthy controls. Increased miR-483-5p levels and decreased miR-20a were further validated in the plasma of 40 myeloma patients and 20 healthy controls using TaqMan quantitative real-time PCR. Receiver operating characteristic (ROC) analysis revealed that miR-483-5p and miR-20a had considerable diagnostic accuracy, yielding the areas under the ROC curve of 0.745 (sensitivity 58%, specificity 90%) and 0.74 (sensitivity 63%, specificity 85%), respectively. Plasma levels of miR-483-5p were associated with ISS staging. Within 14 months of diagnosis, the median progression-free survival of patients with high levels of plasma miR-483-5p was 15 months, in comparison with 21 months for patients with low levels of plasma miR-483-5p (p=0.025). However, miR-20a levels were not correlated with progression-free survival (p>0.05). miR-483-5p has the potential to be a predictor of myeloma survival.

A Reagentless Electrochemiluminescent Immunosensor for Apurinic/apyrimidinic Endonuclease 1 Detection Based on the New Ru(bpy)3(2+)/bi-arginine System

Apurinic/apyrimidinic endonuclease 1 (APE-1), a kind of multifunctional protein widely-distributed in the body, plays an essential role in the DNA base excision repair and serves as multiple possible roles in the response of human cancer to radiotherapy and chemotherapy. In this work, an ultrasensitive solid-state electrochemiluminescence (ECL) immunosensor is designed to determine APE-1 based on the new Ru(bpy)3(2+)/bi-arginine system. The bi-arginine (bi-Arg) is decorated on the Au nanoparticles functionalized magnetic Fe3O4/reduced graphene oxide (bi-Arg/Au@Fe3O4-rGO) according to the self-assembling and covalent cross-linking interaction to obtain the functionalized nanocomposite of bi-Arg/Au@Fe3O4-rGO. Herein, the bi-Arg/Au@Fe3O4-rGO plays not only an amplification label to enhance the ECL signal of Ru(bpy)3(2+) due to the coreactant of bi-Arg but also an ideal nanocarrier to load numerous secondary antibody. Based on sandwich-type immunoassay format, this proposed method offers a linear range of 1.0fgmL(-1)-5.0pgmL(-1) and an estimated detection limit of 0.3fgmL(-1) for the APE-1. Moreover, the reagentless ECL immunosensor also exhibits high sensitivity, excellent selectivity and good stability, which has greatly potential development and application in clinical diagnostics, immunology and biomedical research.

A Risk Evaluation Model of Cervical Cancer Based on Etiology and Human Leukocyte Antigen Allele Susceptibility

There are no reliable risk factors to accurately predict progression to cervical cancer in patients with chronic cervicitis infected with human papillomavirus (HPV). The aim of this study was to create a validated predictive model based on the risk factors for cervical cancer. A model to estimate the risk of cervical cancer may help select patients for intervention therapy in order to reduce the occurrence of cervical cancer after HPV infection.

ElectroTaxis-on-a-Chip (ETC): an Integrated Quantitative High-throughput Screening Platform for Electrical Field-directed Cell Migration

Both endogenous and externally applied electrical stimulation can affect a wide range of cellular functions, including growth, migration, differentiation and division. Among those effects, the electrical field (EF)-directed cell migration, also known as electrotaxis, has received broad attention because it holds great potential in facilitating clinical wound healing. Electrotaxis experiment is conventionally conducted in centimetre-sized flow chambers built in Petri dishes. Despite the recent efforts to adapt microfluidics for electrotaxis studies, the current electrotaxis experimental setup is still cumbersome due to the needs of an external power supply and EF controlling/monitoring systems. There is also a lack of parallel experimental systems for high-throughput electrotaxis studies. In this paper, we present a first independently operable microfluidic platform for high-throughput electrotaxis studies, integrating all functional components for cell migration under EF stimulation (except microscopy) on a compact footprint (the same as a credit card), referred to as ElectroTaxis-on-a-Chip (ETC). Inspired by the R-2R resistor ladder topology in digital signal processing, we develop a systematic approach to design an infinitely expandable microfluidic generator of EF gradients for high-throughput and quantitative studies of EF-directed cell migration. Furthermore, a vacuum-assisted assembly method is utilized to allow direct and reversible attachment of our device to existing cell culture media on biological surfaces, which separates the cell culture and device preparation/fabrication steps. We have demonstrated that our ETC platform is capable of screening human cornea epithelial cell migration under the stimulation of an EF gradient spanning over three orders of magnitude. The screening results lead to the identification of the EF-sensitive range of that cell type, which can provide valuable guidance to the clinical application of EF-facilitated wound healing.

[Modified Bortezomib-based Combination Therapy for Multiple Myeloma]

Influence of Water Content in Mixed Solvent on Surface Morphology, Wettability, and Photoconductivity of ZnO Thin Films

ZnO thin films have been synthesized by means of a simple hydrothermal method with different solvents. The effect of deionized water content in the mixed solvents on the surface morphology, crystal structure, and optical property has been investigated by scanning electron microscopy, X-ray diffraction, and UV-Vis spectrophotometer. A large number of compact and well-aligned hexagonal ZnO nanorods and the maximal texture coefficient have been observed in the thin film, which is grown in the mixed solvent with x = 40%. A lot of sparse, diagonal, and pointed nanorods can be seen in the ZnO thin film, which is grown in the 40-mL DI water solution. The optical band gap decreases firstly and then increases with the increase of x. Reversible wettability of ZnO thin films were studied by home-made water contact angle apparatus. Reversible transition between hydrophobicity and hydrophilicity may be attributed to the change of surface chemical composition, surface roughness and the proportion of nonpolar planes on the surface of ZnO thin films. Photocurrent response of ZnO thin films grown at different solvents were measured in air. The response duration of the thin film, which is grown in the solvent with x = 40%, exhibits a fast growth in the beginning but cannot approach the saturate current value within 100 s. The theoretical mechanism for the slower growth or decay duration of the photocurrent has been discussed in detail.

Highly Enantioselective Phosphination and Hydrophosphonylation of Azomethine Imines: Using Chiral Squaramide As a Hydrogen Bonding Organocatalyst

Enantioselective phosphination and hydrophosphonylation reactions between azomethine imines and diarylphosphine oxides or dialkyl phosphites were respectively developed by the use of a chiral squaramide as the hydrogen bonding organocatalyst, which afforded two types of phosphorus containing product in high yields with good to excellent enantioselectivities.

Impaired Processing Speed and Attention in First-episode Drug Naive Schizophrenia with Deficit Syndrome

Although first-episode drug naive patients with schizophrenia are known to show cognitive impairment, the cognitive performances of these patients, who suffer deficit syndrome, compared with those who suffer non-deficit syndrome is undetermined. The aim of this study was to compare cognitive performances in first-episode drug-naive schizophrenia with deficit syndrome or non-deficit syndrome. First-episode drug naive patients (n=49) and medicated patients (n=108) with schizophrenia, and age, sex, and education matched healthy controls (n=57 for the first-episode group, and n=128 for the medicated group) were enrolled. Patients were divided into deficit or non-deficit syndrome groups, using the Schedule for Deficit Syndrome. Cognitive performance was assessed using the CogState computerized cognitive battery. All cognitive domains in first-episode drug naive and medicated patients showed significant impairment compared with their respective control groups. Furthermore, cognitive performance in first-episode drug naive patients was significantly worse than in medicated patients. Interestingly, the cognitive performance markers of processing speed and attention, in first-episode drug naive patients with deficit syndrome, were both significantly worse than in equivalent patients without deficit syndrome. In contrast, no differences in cognitive performance were found between the two groups of medicated patients. In conclusion, this study found that first-episode drug naive schizophrenia with deficit syndrome showed significantly impaired processing speed and attention, compared with patients with non-deficit syndrome. These findings highlight processing speed and attention as potential targets for pharmacological and psychosocial interventions in first-episode schizophrenia with deficit syndrome, since these domains are associated with social outcomes.

[Progress on Association Between COMT Gene and Violence Behavior in Patients with Schizophrenia]

The prevalence of violence behavior in patients with schizophrenia is higher than that in common population. Data suggest that genetic factors may play a substantial role for the etiology of the behavior. Among the particular gene polymorphisms that have been considered to be involved in violence behavior, the catechol-O-methyltransferase (COMT) gene had been the focus of recent research. This article reviews the association research between COMT gene and violence behavior in patients with schizophrenia in several aspects: SNP polymorphism of COMT Val158Met and COMT Ala72Ser, haplotype of COMT gene and DNA methylation of promoter region of COMT gene. The genetic research direction is presented for patients with schizophrenia.

Neuropeptides Encoded by the Genomes of the Akoya Pearl Oyster Pinctata Fucata and Pacific Oyster Crassostrea Gigas: a Bioinformatic and Peptidomic Survey

Oysters impart significant socio-ecological benefits from primary production of food supply, to estuarine ecosystems via reduction of water column nutrients, plankton and seston biomass. Little though is known at the molecular level of what genes are responsible for how oysters reproduce, filter nutrients, survive stressful physiological events and form reef communities. Neuropeptides represent a diverse class of chemical messengers, instrumental in orchestrating these complex physiological events in other species.

A BHLH Transcription Factor Regulates Iron Intake Under Fe Deficiency in Chrysanthemum

Iron (Fe) deficiency can represent a serious constraint on crop growth and productivity. A number of members of the bHLH transcription factor family are known to be involved in the plant Fe deficiency response. Plants have evolved two distinct uptake strategies when challenged by Fe deficiency: dicotyledonous and non-graminaceous species rely mostly on a reduction strategy regulated by bHLH transcription factors, whereas rice relies on a chelation strategy, also regulated by bHLH transcription factors. CmbHLH1, a bHLH transcription factor which is localized within the nucleus, was isolated from chrysanthemum. Its transcription was up-regulated both by Fe deficiency and by the exogenous application of abscisic acid. The roots of transgenic chrysanthemum plants in which CmbHLH1 was up-regulated were better able than those of the wild type chrysanthemum cultivar to acidify their immediate external environment by enhancing the transcription of the H(+)-ATPase encoding gene CmHA. However, there was no effect of the transgene on the efficiency of uptake of either manganese or zinc. Here, Chrysanthemum CmbHLH1 contributed to Fe uptake via H(+)-ATPase mediated acidification of the rhizosphere. ABA may be positively involved in the process.

Semaphorin 3A Blocks the Formation of Pathologic Choroidal Neovascularization Induced by Transforming Growth Factor Beta

Choroidal neovascularization (CNV) is a major cause of vision loss in retinal diseases such as age-related macular degeneration (AMD). Previously, we demonstrated that semaphorin3A (Sema3A), which is a chemorepellent guidance molecule, inhibited the formation of retina neovascularization. In the present study, we investigated the antiangiogenic effects of Sema3A on transforming growth factor beta (TGF-β) in vitro and in vivo.

Anti-angiogenic Effects of a Mutant Endostatin: a New Prospect for Treating Retinal and Choroidal Neovascularization

Pathological fundus angiogenesis is a major cause of vision loss in retina diseases. Endostatin, a C-terminal fragment of collagen XVIII, is an endogenous anti-angiogenic protein. The present study aimed to investigate the in vitro and in vivo anti-angiogenic properties of two proteins: an N-terminal H1D/H3D mutant endostatin (M-ES) and a polyethylene glycol propionaldehyde (PEG) covalent M-ES (PEG-M-ES).

Determination of Metabolites of Diosmetin-7-O-glucoside by a Newly Isolated Escherichia Coli from Human Gut Using UPLC-Q-TOF/MS

Different human intestinal bacteria were isolated and screened for their ability to transform diosmetin-7-O-glucoside. A Gram-negative anaerobic bacterium, strain 4, capable of metabolizing diosmetin-7-O-glucoside was newly isolated. Its 16S rRNA gene sequence displayed 99% similarity with that of Escherichia. Then strain 4 was identified as a species of the genus Escherichia and was named Escherichia sp. 4. Additionally, an ultraperformance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) technique combined with Metabolynx software method was established to screen the metabolites of diosmetin-7-O-glucoside. Comparing the retention time and MS/MS spectrum, three metabolites were detected and tentatively identified. These metabolites were acquired by four proposed metabolic pathways including dehydroxylation, deglycosylation, methylation, and acetylation. Diosmetin-7-O-glucoside was mainly bioconverted to considerable amounts of diosmetin and minor amounts of acacetin by the majority of the isolated intestinal bacteria such as Escherichia sp. 4. Subsequently, several strains could degrade acacetin to produce methylated and acetylated acacetin. The metabolites and metabolic pathways of diosmetin-7-O-glucoside by human intestinal bacterium Escherichia sp. 4 were first investigated.

Treatment Adherence and Health Outcomes in MSM with HIV/AIDS: Patients Enrolled in "one-stop" and Standard Care Clinics in Wuhan China

Conducted in Wuhan China, this study examined follow-up and health markers in HIV patients receiving care in two treatment settings. Participants, all men who have sex with men, were followed for 18-24 months.

Chemical Composition, Antimicrobial and Anti-inflammatory Activities of the Essential Oil from Maqian (Zanthoxylum Myriacanthum Var. Pubescens) in Xishuangbanna, SW China

Maqian (Zanthoxylum myriacanthum var. pubescens Huang) is widely consumed as an indigenous remedy for digestive disorders, detoxification, detumescence and analgesia by the ethnic groups in Xishuangbanna, SW China. A related species, Huajiao (Zanthoxylum schinifolium Sieb. et Zucc.), has similar uses in traditional Chinese medicine. We aimed to scientifically validate the traditional uses by investigating and comparing the chemical composition, antimicrobial and anti-inflammatory activities of the essential oils of Maqian and Huajiao.

TRPM7 is Required for Ovarian Cancer Cell Growth, Migration and Invasion

Our previous study demonstrated that the melastatin-related transient receptor potential channel 7 (TRPM7) was highly expressed in ovarian carcinomas and its overexpression was significantly associated with poor prognosis in ovarian cancer patients. However, the function of TRPM7 in ovarian cancer is mostly unknown. In this study, we examined the roles of TRPM7 in ovarian cancer cell proliferation, migration and invasion. We found that short hairpin RNA interference-mediated silence of TRPM7 significantly inhibited cell proliferation, colony formation, migration and invasion in multiple ovarian cancer cell lines. Mechanistic investigation revealed that silence of TRPM7 decreased phosphorylation levels of Akt, Src and p38 and increased filamentous actin and focal adhesion number in ovarian cancer cells. Thus, our results suggest that TRPM7 is required for proliferation, migration and invasion of ovarian cancer cells through regulating multiple signaling transduction pathways and the formation of focal adhesions.

The Efficacy of Trimetazidine on Stable Angina Pectoris: a Meta-analysis of Randomized Clinical Trials

This meta-analysis aimed to evaluate the efficacy of trimetazidine in combination with other anti-anginal drugs versus other anti-anginal drugs in the treatment of stable angina pectoris (SAP). Randomized controlled trials (RCTs) published in English and Chinese were retrieved from computerized databases: Embase, PubMed, and CNKI. Primary outcomes consist of clinical parameters (numbers of weekly angina attacks and nitroglycerin use) and ergometric parameters (time to 1mm ST-segment depression, and total work (in Mets) and exercise duration (in seconds) at peak exercise) in stable angina pectoris treated by trimetazidine or not. The quality of studies was evaluated using Jadad score. Data analysis of 13 studies was performed using Stata 12.0 software. Results showed that treatment of trimetazidine and other anti-anginal drugs was associated with a smaller weekly mean number of angina attacks (WMD=-0.95, 95%CI: -1.30 to -0.61, Z=5.39, P<0.001), fewer weekly nitroglycerin use (WMD=-0.98, 95%CI: -1.44 to -0.52, Z=4.19, P<0.001), longer time to 1mm ST-segment depression (WMD=0.30, 95%CI: 0.17 to 0.43, Z=4.46, P<0.001), higher total work (WMD=0.82, 95%CI: 0.44 to 1.20, Z=4.22, P<0.001) and longer exercise duration at peak exercise (WMD=49.81, 95%CI: 15.04 to 84.57, Z=6.38, P<0.001) than treatment of other anti-anginal drugs for stable angina pectoris. Sensitivity analysis was performed. Sub-group analysis showed that treatment duration was not a significant moderator and patients treated within 8 weeks and above 12 weeks had no difference in the outcomes addressed in this meta-analysis. No publish bias was detected. This meta-analysis confirms the efficacy of trimetazidine in the treatment of stable angina pectoris, in comparison with conventional antianginal agents, regardless of treatment duration.

Diterpenes from a Chinese Collection of the Brown Alga Dictyota Plectens

Twenty-seven diterpenes of six chemical classes, including seven new diterpenes (1, 2, 6, 10, 11, 16, and 19), have been isolated from a collection of the brown alga Dictyota plectens from the South China Sea. The structures of the new diterpenes were elucidated by extensive spectroscopic analysis and by comparison with reported data. In the in vitro assays, 9, 12, 14, 16, and 22 showed inhibitory activity against HIV-1 replication with IC50 values of 16.1-30.5 μM, compounds 5, 13, 24, and 26 exhibited anti-H5N1 activity with inhibition rates of 50%-62% at 30.0 μM, and 12 and 24 also showed potent inhibition against LPS-induced NO production with inhibition rates of 90% and 86%, respectively, at 10.0 μM.

Identification of Differentially-expressed Genes in Intestinal Gastric Cancer by Microarray Analysis

Gastric cancer (GC) is one of the most frequent malignant tumors. In order to systematically characterize the cellular and molecular mechanisms of intestinal GC development, in this study, we used 22K oligonucleotide microarrays and bioinformatics analysis to evaluate the gene expression profiles of GC in 45 tissue samples, including 20 intestinal GC tissue samples, 20 normal appearing tissues (NATs) adjacent to tumors and 5 noncancerous gastric mucosa tissue samples. These profiles allowed us to explore the transcriptional characteristics of GC and determine the change patterns in gene expression that may be of clinical significance. 1519 and 1255 differentially-expressed genes (DEGs) were identified in intestinal GC tissues and NATs, respectively, as determined by Bayesian analysis (P<0.001). These genes were associated with diverse functions such as mucosa secretion, metabolism, proliferation, signaling and development, which occur at different stages of GC development.

The Predictive Value of the Boston Acute Stroke Imaging Scale (BASIS) in Acute Ischemic Stroke Patients Among Chinese Population

Evaluate the predictive value of Boston Acute Stroke Imaging Scale (BASIS) in acute ischemic stroke in Chinese population.

Investigation on Reference Intervals and Regional Differences of Platelet Indices in Healthy Chinese Han Adults

Reference intervals are important for interpretation of clinical laboratory tests. The platelet (PLT) indices such as the mean platelet volume (MPV) and platelet distribution width (PDW) are newer hematological parameters, which have been recently reported as clinically valuable biomarkers. However, there are not many studies that have estimated the reference intervals for these parameters in healthy Chinese Han adults.

Biogenic Magnetic Nanoparticles from Burkholderia Sp. YN01 Exhibiting Intrinsic Peroxidase-like Activity and Their Applications

A novel bacterial strain containing biogenic magnetic nanoparticles (BMNPs) was isolated from the sediments of Songhua River in Harbin, China, and was identified as Burkholderia sp. YN01. Extracted BMNPs from YN01 were characterized as pure face-centered cubic Fe3O4 with an average size of 80 nm through transmission electron microscope (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The hysteresis parameters of the BMNP samples such as Bc and Bcr and ratios Mrs/Ms were deduced as 35.6 mT, 43.2 mT, and 0.47, respectively, indicating that the BMNPs exhibit a ferromagnetic behavior. This is the first report concerning on biogenic Fe3O4 NPs produced in Burkholderia genus. Significantly, the BMNPs were proved to possess intrinsic peroxidase-like activity that could catalyze the oxidation of peroxidase substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of H2O2. Kinetic analysis indicates that the catalytic behavior is in accord with typical Michaelis-Menten kinetics and follows ping-pong mechanism. The catalytic constants (K cat) were 6.5 × 10(4) s(-1) and 0.78 × 10(4) s(-1) with H2O2 and TMB as substrate, respectively, which was higher than that of horseradish peroxidase (HRP). Electron spin resonance (ESR) spectroscopy experiments showed that the BMNPs could catalyze H2O2 to produce hydroxyl radicals. The origin of peroxidase-like activity is also associated with their ability to transfer electron between electrode and H2O2 according to an electrochemical study. As a novel peroxidase mimetic, the BMNPs were employed to offer a simple, sensitive, and selective colorimetric method for H2O2 and glucose determination, and the BMNPs could efficiently catalyze the degradation of phenol and Congo red dye.

Association of HLA-DP/DQ, STAT4 and IL-28B Variants with HBV Viral Clearance in Tibetans and Uygurs in China

Several genome-wide association studies have revealed that HLA-DP/DQ, STAT4 and IL-28B associated with liver diseases. But because of population heterogeneity, different races would have different causative polymorphisms. Therefore, in this study, we included Chinese Tibetans and Uygurs to examine the roles of these genes on HBV natural clearance.

Highly Efficient Electrogenerated Chemiluminescence Quenching of PEI Enhanced Ru(bpy)₃²⁺ Nanocomposite by Hemin and Au@CeO₂ Nanoparticles

In this work, a new signal amplified strategy based on the quenching effect of hemin and Au nanoparticles decorated CeO2 nanoparticles (Au@CeO2 NPs) for ultrasensitive detection of thrombin (TB) is reported for the first time. Herein, the poly(ethylenimine) (PEI) enhanced Ru(bpy)3(2+) nanocomposite was implemented by direct chemical polymerization, which could provide the desirable enhanced initial ECL signal. Furthermore, the detection aptamer of thrombin (TBA 2) was immobilized on Au@CeO2 NPs to form TBA 2/Au@CeO2 conjugates. Then, the G-rich DNA of TBA 2 sequence could fold into a G-quadruplex structure to embed hemin to obtain the quenching probe of hemin/TBA 2/Au@CeO2 conjugates. In the presence of target TB, the sandwiched structure could be formed between capture aptamer (TBA 1), TB and hemin/TBA 2/Au@CeO2 conjugates, thereby resulting in a proportional quenching in ECL response with TB, due to the quenching of both hemin and Au@CeO2 NPs. As a result, the signal-off aptasensor showed a wider linear range response from 10(-13) to 10(-8) M with lower detection limit of 0.03 pM.

Size-dependent Nonlinear Optical Properties of Atomically Thin Transition Metal Dichalcogenide Nanosheets

Size-dependent nonlinear optical properties of modification-free transition metal dichalcogenide (TMD) nanosheets are reported, including MoS2 , WS2 , and NbSe2 . Firstly, a gradient centrifugation method is demonstrated to separate the TMD nanosheets into different sizes. The successful size separation allows the study of size-dependent nonlinear optical properties of nanoscale TMD materials for the first time. Z-scan measurements indicate that the dispersion of MoS2 and WS2 nanosheets that are 50-60 nm thick leads to reverse saturable absorption (RSA), which is in contrast to the saturable absorption (SA) seen in the thicker samples. Moreover, the NbSe2 nanosheets show no size-dependent effects because of their metallic nature. The mechanism behind the size-dependent nonlinear optical properties of the semiconductive TMD nanosheets is revealed by transient transmission spectra measurements.

Downregulation of 425G>a Variant of Calcium-binding Protein S100A14 Associated with Poor Differentiation and Prognosis in Gastric Cancer

Altered level of S100 calcium-binding proteins is involved in tumor development and progression. However, their role in gastric cancer (GC) is not well documented. We investigated the expression pattern of S100 proteins and differentiation or prognosis as well as possible mechanisms in GC.

Correlation of IL-1F Genetic Polymorphisms with the Risk of Colorectal Cancer Among Chinese Populations

Inflammatory/immune cells have the power of infiltrating almost all human solid tumors and influencing all stages of carcinogenesis because of their stimulation of various cytokine subsets. This study aims to determine the correlation of single nucleotide polymorphisms in the IL-17F gene and the risk of colorectal cancer (CRC). One thousand patients diagnosed with CRC and a control group of 354 healthy controls were involved. Peripheral blood samples were collected. The PCR-RFLP method was used to detect the 7383A>G (rs2397084) and 7488T>C (rs763780) in the IL-17F gene. Statistical analyses were conducted with version 12.0 STATA statistical software. We found that the allele model suggested that patients carrying C allele were 1.67 times more likely to develop CRC than healthy controls (odds ratio (OR) = 1.67, 95% confidence interval (CI) = 1.22-2.27, P = 0.001). Similarly, the homozygous and dominant models also revealed that the minor IL-17F 7488C allele conferred an increased CRC risk compared to the major T allele among our study participants (CC vs. TT: OR = 4.15, 95% CI = 1.26-13.36, P = 0.011; TC+CC vs. TT: OR = 1.46, 95% CI = 1.04-2.05, P = 0.027). However, all genetic models indicated that the IL-17F 7383A>G (rs2397084) polymorphism was not associated with CRC risk (all P > 0.05). The results of this study indicate that the 7488T>C (rs763780) in the IL-17F gene may be correlated with increased risk of CRC.

Biomimetic Stochastic Topography and Electric Fields Synergistically Enhance Directional Migration of Corneal Epithelial Cells in a MMP-3-dependent Manner

Directed migration of corneal epithelial cells (CECs) is critical for maintenance of corneal homeostasis as well as wound healing. Soluble cytoactive factors and the intrinsic chemical attributes of the underlying extracellular matrix (ECM) participate in stimulating and directing migration. The central importance of the intrinsic biophysical attributes of the microenvironment of the cell in modulating an array of fundamental epithelial behaviors including migration has been widely documented. Among the best measures of these attributes are the intrinsic topography and stiffness of the ECM and electric fields (EFs). How cells integrate these multiple simultaneous inputs is not well understood. Here, we present a method that combines the use of (i) topographically patterned substrates (mean pore diameter 800nm) possessing features that approximate those found in the native corneal basement membrane; and (ii) EFs (0-150mVmm(-1)) mimicking those at corneal epithelial wounds that the cells experience in vivo. We found that topographic cues and EFs synergistically regulated directional migration of human CECs and that this was associated with upregulation of matrix metalloproteinase-3 (MMP3). MMP3 expression and activity were significantly elevated with 150mVmm(-1) applied-EF while MMP2/9 remained unaltered. MMP3 expression was elevated in cells cultured on patterned surfaces against planar surfaces. The highest single-cell migration rate was observed with 150mVmm(-1) applied EF on patterned and planar surfaces. When cultured as a confluent sheet, EFs induced collective cell migration on stochastically patterned surfaces compared with dissociated single-cell migration on planar surfaces. These results suggest significant interaction of biophysical cues in regulating cell behaviors and will help define design parameters for corneal prosthetics and help to better understand corneal wound healing.

Subconjunctival Injection of XG-102, a C-Jun N-terminal Kinase Inhibitor Peptide, in the Treatment of Endotoxin-induced Uveitis in Rats

XG-102, a TAT-coupled dextrogyre peptide inhibiting the c-Jun N-terminal kinase, was shown efficient in the treatment of experimental uveitis. Preclinical studies are now performed to determine optimal XG-102 dose and route of administration in endotoxin-induced uveitis (EIU) in rats with the purpose of clinical study design.

Effect of Quinoid Redox Mediators on the Aerobic Decolorization of Azo Dyes by Cells and Cell Extracts from Escherichia Coli

It is widely accepted that the addition of redox mediators increases the decolorization rates of azo dyes by bacterial strains under anaerobic conditions. However, little information exists about whether quinoid redox mediators can enhance the performance of aerobic azo dye decolorization. In the present study, quinone-mediated decolorization of different azo dyes by whole cells and cell extracts from the Escherichia coli strain CD-2 under aerobic conditions were investigated. The results demonstrated that reduction rates of different azo dyes were greatly increased when quinone compounds were used as redox mediators. Compared with menadione, 2-hydroxy-1,4-naphthoquinone (lawsone) was more effective at aiding azo dye degradation and the optimum concentration for lawsone is 0.1 mM. Strain CD-2 and the anthraquinone were co-immobilized by entrapment in different polymeric matrices. The co-immobilized beads exhibited good catalytic activity for azo dye degradation and kept stable during successive repeated experiments. The mechanism of the quinone-mediated reduction showed that although whole cells incubated with quinones could significantly increase the rate of decolorization of azo dyes, the quinone compounds did not directly promote azoreductase activity. According to the survey, this is the first report to confirm that the addition of quinoid redox mediators to bacteria increased decolorization under aerobic conditions.

NLRP3 Inflammasome Activation is Essential for Paraquat-induced Acute Lung Injury

The innate immune response is important in paraquat-induced acute lung injury, but the exact pathways involved are not elucidated. The objectives of this study were to determine the specific role of the NLRP3 inflammasome in the process. Acute lung injury was induced by administering paraquat (PQ) intraperitoneally. NLRP3 inflammasome including NLRP3, ASC, and caspase-1 mRNA and protein expression in lung tissue and IL-1β and IL-18 levels in BALF were detected at 4, 8, 24, and 72 h after PQ administration in rats. Moreover, rats were pretreated with 10, 30, and 50 mg/kg NLRP3 inflammasome blocker glybenclamide, respectively, 1 h before PQ exposure. At 72 h after PQ administration, lung histopathology changes, NLRP3, ASC, and caspase-1 protein expression, as well as secretion of cytokines including IL-1β and IL-18 in BALF were investigated. The NLRP3 inflammasome including NLRP3, ASC, caspase-1 expression, and cytokines IL-1β and IL-18 levels in PQ poisoning rats were significantly higher than that in the control group. NLRP3 inflammasome blocker glybenclamide pretreatment attenuated lung edema, inhibited the NLRP3, ASC, and caspase-1 activation, and reduced IL-1β and IL-18 levels in BALF. In the in vitro experiments, IL-1β and IL-18 secreted from RAW264.7 mouse macrophages treated with paraquat were attenuated by glybenclamide. In conclusion, paraquat can induce IL-1β/IL-18 secretion via NLRP3-ASC-caspase-1 pathway, and the NLRP3 inflammasome is essential for paraquat-induced acute lung injury.

Transplantation of Olfactory Ensheathing Cells Attenuates Acute Carbon Monoxide Poisoning-induced Brain Damages in Rats

In this study, the therapeutic effect of olfactory ensheathing cells (OEC) transplantation on brain damage was evaluated on acute carbon monoxide (CO) poisoning rat model. Two weeks after primary culture, OECs were microinjected into hippocampus of CO poisoning rats. Survival of OECs in the host was observed and quantified. OECs survived at 2 weeks, but surviving cell number was found sharply decreased at 6 weeks and reduced to less than 10(3) at 8 weeks after transplantation. At 2 weeks after transplantation, motor function test and cerebral edema assay were performed and followed by pathological examination including hematoxylin and eosin and immunohistochemistry staining to observe the neuron injury and synapsin I and growth associated protein-43 (GAP-43) expression. Furthermore, biomarkers of oxidative stress and apoptosis related proteins in the hippocampus were detected. The results showed that CO exposure led to neurological dysfunction and cerebral edema in rats. After OEC transplantation, neurological function was significantly improved and the cerebral edema was alleviated. In addition, the numbers of neurons and Nissl bodies were increased and synapsin I and GAP-43 protein expressions were upregulated in the hippocampus. Compared with CO poisoned rats, superoxide dismutase activity and glutathione content were both increased and methane dicarboxylic aldehyde level was decreased in the hippocampus of OEC transplanted rats. Moreover, OEC transplantation reduced apoptosis induced by CO exposure. The Bcl-2 expression was significantly upregulated and Bax expression was significantly downregulated. The activity of caspase-3 and the cleaved-poly ADP-ribose polymerase expression were decreased. Taken together, our data suggest that OEC attenuates brain damages induced by acute CO poisoning within 2 weeks after transplantation.

Proinflammatory Secreted Phospholipase A2 Type IIA (sPLA-IIA) Induces Integrin Activation Through Direct Binding to a Newly Identified Binding Site (site 2) in Integrins αvβ3, α4β1, and α5β1

Integrins are activated by signaling from inside the cell (inside-out signaling) through global conformational changes of integrins. We recently discovered that fractalkine activates integrins in the absence of CX3CR1 through the direct binding of fractalkine to a ligand-binding site in the integrin headpiece (site 2) that is distinct from the classical RGD-binding site (site 1). We propose that fractalkine binding to the newly identified site 2 induces activation of site 1 though conformational changes (in an allosteric mechanism). We reasoned that site 2-mediated activation of integrins is not limited to fractalkine. Human secreted phospholipase A2 type IIA (sPLA2-IIA), a proinflammatory protein, binds to integrins αvβ3 and α4β1 (site 1), and this interaction initiates a signaling pathway that leads to cell proliferation and inflammation. Human sPLA2-IIA does not bind to M-type receptor very well. Here we describe that sPLA2-IIA directly activated purified soluble integrin αvβ3 and transmembrane αvβ3 on the cell surface. This activation did not require catalytic activity or M-type receptor. Docking simulation predicted that sPLA2-IIA binds to site 2 in the closed-headpiece of αvβ3. A peptide from site 2 of integrin β1 specifically bound to sPLA2-IIA and suppressed sPLA2-IIA-induced integrin activation. This suggests that sPLA2-IIA activates αvβ3 through binding to site 2. sPLA2-IIA also activated integrins α4β1 and α5β1 in a site 2-mediated manner. We recently identified small compounds that bind to sPLA2-IIA and suppress integrin-sPLA2-IIA interaction (e.g. compound 21 (Cmpd21)). Cmpd21 effectively suppressed sPLA2-IIA-induced integrin activation. These results define a novel mechanism of proinflammatory action of sPLA2-IIA through integrin activation.

Ultra-performance Liquid Chromatography Coupled with Quadrupole Time-of-flight Mass Spectrometry for Rapid Analysis of the Metabolites of Morroniside Produced by Human Intestinal Bacteria

Morroniside, the most abundant iridoid glycoside in the valuable traditional Chinese medicine Fructus Corni, exhibits various pharmacological activities and biological effects. Intestinal flora plays an important role in the metabolism of drug compounds, which might lead to the variation of ethnopharmacological profile of the medicine. However, little is known of the interactions of the morroniside with human intestinal bacteria. In this study, different pure bacteria were isolated from human feces and their capability to convert morroniside were investigated. The metabolites of morroniside were analyzed by ultra high performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) technique using Metabolynx™ software. Parent compound and three metabolites were detected and tentatively identified based on the characteristics of their protonated ions. The parent is proposed to be metabolized by three main metabolic pathways including deglycosylation, dehydroxylation and methylation. Morroniside was firstly metabolized to its aglycone (M1), and then was further converted to dehydroxylated aglycone (M2) and methylated aglycone (M3). This is the first report of the metabolism of morroniside by human intestinal bacteria. These metabolites might influence the biological activities of morroniside in vivo, which could affect the clinical effects of medicines. Thus, the study on the metabolism of morroniside by human intestinal bacteria is very helpful to unravel how traditional medicines work.

A Porous Metal-organic Framework Containing Multiple Active Cu(2+) Sites for Highly Efficient Cross Dehydrogenative Coupling Reaction

A novel 3D porous metal-organic framework was constructed from imidazole carboxylate linkers and copper(ii) nodes, which in situ generates multiple active Cu(II) sites in the nanosized channel walls for highly efficient cross dehydrogenative coupling reaction between 1,2,3,4-tetrahydroisoquinoline derivatives and nitroalkanes that are superior to the simple copper salts.

Club Drug Use and Associated High-risk Sexual Behaviour in Six Provinces in China

To investigate the characteristics of the different club drugs and associated high-risk sexual behaviours in China.

Tumor Necrosis Factor-α-Induced Protein-8 Like-2 (TIPE2) Upregulates P27 to Decrease Gastic Cancer Cell Proliferation

The pathogenesis of gastric cancer is not completely understood. Tumor necrosis factor-α-induced protein-8 like-2 (TIPE2) has recently been identified as a novel negative regulator gene of the immune system, and studies in mice and humans have suggested its inhibitory action in both inflammation and cancer. In this study, we examined the expression levels of TIPE2 in human gastric cancer tissues and also samples of paraneoplastic control tissue, and found that TIPE2 expression was reduced in gastric cancer. To investigate the role of TIPE2 in gastric cell carcinogenesis, a TIPE2 plasmid was introduced into gastric cell lines and TIPE2 function was examined. Colony-forming assays showed that restoration of TIPE2 expression in gastric cells significantly suppressed cell proliferation. Analysis by flow cytometry showed that the number of cells in the S phase of the cell cycle was reduced concomitant with TIPE2 expression, and cell apoptosis was maintained at a low level. Microarray and western blot analyses revealed that TIPE2 selectively up-regulated N-ras and p27 expression. The role of p27 in mediating TIPE2-associated cell growth inhibition was verified by a p27 siRNA interference assay. In this study, we proved that TIPE2 is an inhibitor of gastric cancer cell growth, and suggest that TIPE2 might promote a p27-associated signaling cascade that leads to restored control of the cell cycle and cell division. Our results provide a new molecular mechanism by which TIPE2 may regulate proliferation of gastric cells. J. Cell. Biochem. 116: 1121-1129, 2015. © 2015 Wiley Periodicals, Inc.

Delayed Hyperoxic Ventilation Attenuates Oxygen-induced Free Radical Accumulation During Early Reperfusion After Global Brain Ischemia

To compare the effect of immediate and delayed administration of oxygen on the accumulation of free radicals in ischemia-reperfusion animal models. Thirty-two adult male Mongolian gerbils with microdialysis probes implanted in the right hippocampal CA1 were divided randomly into four groups (eight each). One group was sham-operated (Sham group) whereas the other three groups were subjected to 10 min bilateral carotid artery occlusion (BCAO). BCAO-treated animals were then subjected to the following: (a) immediate 30% O2 (near normoxia, NO group), (b) immediate 100% O2 (hyperoxia, HO group), and (c) 30% O2 for 60 min, followed by 100% O2 for 60 min (delayed hyperoxia, DHO group). Hippocampal accumulation of hydroxyl radicals (•OH) during reperfusion was estimated by measuring 2,3-dihydroxybenzoic acid (DHBA) and 2,5-DHBA in microdialysis perfusate. Hippocampi were removed 2 h after perfusion to measure malondialdehyde, pyruvate dehydrogenase activity, indices of lipid peroxidation, and cellular respiration. At 24 h after BCAO, the histology of hippocampi was analyzed to rate the injury. Immediately after the onset of reperfusion, all groups showed markedly elevated DHBA, which returned to baseline over 1-2 h. Compared with the NO group, the HO group showed significantly higher peak DHBA and slower recovery. In contrast, the DHO group was not significantly different from the NO group in terms of the DHBA level. DHO animals also showed significantly lower hippocampal malondialdehyde accumulation and higher pyruvate dehydrogenase activity at 2 h after reperfusion versus the HO group. Histology analysis also showed animals in the DHO group with ameliorated injury compared with the HO group. Hydroxyl radical accumulation was more sensitive to O2 during early reperfusion. Delayed hyperoxia may re-establish oxidative metabolism while minimizing oxidative stress after CA.

CD44v6 Promotes β-catenin and TGF-β Expression, Inducing Aggression in Ovarian Cancer Cells

A high expression of CD44v6 has been reported in numerous malignant cancers, including stomach, prostate, lung and colon. However, the pathological role and the regulatory mechanisms of CD44v6 have yet to be elucidated. In the present study, the expression levels of CD44v6 were shown to be significantly higher in ovarian cancer tissues, as compared with adjacent normal tissues. Furthermore, the upregulated expression levels of CD44v6 were correlated with disease recurrence and poor survival in patients. The expression of CD44v6 was knocked down in the CAOV3 ovarian cell line, by transfection of a specific small hairpin RNA. The present study showed a correlation between the aggression, viability, invasion and migration of the ovarian cancer cells, with the expression of CD44v6. In addition, the expression of CD44v6 was positively correlated with the expression levels of β‑catenin and tumor growth factor‑β, which indicates that the effects of CD44v6 on ovarian cancer cell aggression may be mediated by these two signaling pathways. In conclusion, the present study provides a novel insight into the association between CD44v6 expression and ovarian cancer. CD44v6 may provide a novel target for the prognosis and treatment of ovarian cancer.

Mechanism-based Inhibitory and Peroxisome Proliferator-activated Receptor α-dependent Modulating Effects of Silybin on Principal Hepatic Drug-metabolizing Enzymes

Silybin, a major pharmacologically active compound in silymarin, has been widely used in combination with other prescriptions in the clinic to treat hepatitis and a host of other diseases. Previous studies suggested that silybin is a potential inhibitor of multiple drug-metabolizing enzymes (DMEs); however, the in vitro to in vivo translation and the mechanisms involved remain established. The aim of this study was to provide a mechanistic understanding of the regulatory effects of silybin on principal DMEs. Silybin (50 or 150 mg/kg/d) was administered to mice for a consecutive 14 days. The plasma and hepatic exposure of silybin were detected; the mRNA, protein levels, and enzyme activities of principal DMEs were determined. The results demonstrated that the enzyme activities of CYP1A2, CYP2C, CYP3A11, and UGT1A1 were significantly repressed, whereas little alteration of the mRNA and protein levels was observed. Silybin inhibits these DMEs in a mechanism-based and/or substrate-competitive manner. More importantly, silybin was found to be a weak agonist of peroxisome proliferator-activated receptor (PPAR)α, as evidenced from the molecular docking, reporter gene assay, and the targeting gene expression analysis. However, silybin could significantly compromise the activation of PPARα by fenofibrate, characterized with significantly repressed expression of PPARα targeting genes, including L-FABP, ACOX1, and UGT1A6. This study suggests that silybin, despite its low bioavailability, may inhibit enzyme activities of multiple DMEs in a mechanism-based mode, and more importantly, may confer significant drug-drug interaction with PPARα agonists via the repression of PPARα activation in a competitive mode.

Generation of Hydrate Forms of Paroxetine HCl from the Amorphous State: an Evaluation of Thermodynamic and Experimental Predictive Approaches

In this study, we evaluate the use of theoretical thermodynamic analysis of amorphous paroxetine hydrochloride (HCl) as well as experimental assessment in order to identify the most promising approach to stability and dissolution behaviour prediction, particularly in relation to stoichiometric and nonstoichiometric hydrate formation. Differential scanning calorimetry, thermogravimetric analysis, Fourier transform infrared and X-ray diffraction techniques were used. Parameters including heat capacity, configurational thermodynamic quantities, fragility and relaxation time classified amorphous paroxetine HCl as a moderate fragile glass with a considerable degree of molecular mobility. Solubility studies indicated little advantage of the amorphous form over the crystalline due to conversion to the hydrate Form I during equilibration, while the dissolution rate was higher for the amorphous form under sink conditions. A marked difference in the physical stability of amorphous paroxetine HCl was observed between dry and low humidity storage, with the system recrystallizing to the hydrate form. We conclude that, in this particular case (amorphous conversion to the hydrate), water may be playing a dual role in both plasticizing the amorphous form and driving the equilibrium towards the hydrate form, hence prediction of recrystallization behaviour from amorphous characteristics may be confounded by the additional process of hydrate generation.

Rare Variants in RTEL1 Are Associated with Familial Interstitial Pneumonia

Up to 20% of cases of idiopathic interstitial pneumonia cluster in families, comprising the syndrome of familial interstitial pneumonia (FIP); however, the genetic basis of FIP remains uncertain in most families.

Skin Squamous Cell Carcinoma Presenting As Cellulitis

In general, skin squamous cell carcinoma (SCC) presents as papules or plaques with erythematous or pigmented appearance that may ulcerate the skin. Cellulitis caused by metastatic deposit from a known primary skin SCC has been reported once.1 We describe a patient who presented with cellulitis on the face that did not respond well to full course of antibiotics treatment, and turned out to be a newly diagnosed SCC after biopsy. Other differential diagnoses, such as malignancy, should be suspected in all unusual presentations and biopsy should be taken if patients do not show an optimal and desired improvement after receiving a full-course of antibiotic therapy for cellulitis.

The Effectiveness and Safety of a Danshen-containing Chinese Herbal Medicine for Diabetic Retinopathy: A Randomized, Double-blind, Placebo-controlled Multicenter Clinical Trial

Salvia miltiorrhiza (Danshen in Chinese) is a common traditional Chinese herbal medicine often used to treat many medical conditions. The Compound Danshen Dripping Pill (CDDP) is a danshen-containing Chinese herbal product for the treatment of cardiovascular diseases. However, to date, no controlled clinical studies have been conducted to evaluate the effects of CDDP on diabetic retinopathy (DR).

Quercetin Inhibits Vascular Endothelial Growth Factor-induced Choroidal and Retinal Angiogenesis in Vitro

The aim of this study was to investigate the effects of quercetin on vascular endothelial growth factor (VEGF)-induced choroidal and retinal angiogenesis in vitro using a rhesus macaque choroid-retinal endothelial (RF/6A) cell line.

Novel Covalent Modification of Human Anaplastic Lymphoma Kinase (ALK) and Potentiation of Crizotinib-mediated Inhibition of ALK Activity by BNP7787

BNP7787 (Tavocept, disodium 2,2'-dithio-bis-ethanesulfonate) is a novel, investigational, water-soluble disulfide that is well-tolerated and nontoxic. In separate randomized multicenter Phase II and Phase III clinical trials in non-small-cell lung cancer (NSCLC) patients, treatment with BNP7787 in combination with standard chemotherapy resulted in substantial increases in the overall survival of patients with advanced adenocarcinoma of the lung in the first-line treatment setting. We hypothesized that BNP7787 might interact with and modify human anaplastic lymphoma kinase (ALK). At least seven different variants of ALK fusions with the gene encoding the echinoderm microtubule-associated protein-like 4 (EML4) are known to occur in NSCLC. EML4-ALK fusions are thought to account for approximately 3% of NSCLC cases. Herein, we report the covalent modification of the kinase domain of human ALK by a BNP7787-derived mesna moiety and the functional consequences of this modification in ALK assays evaluating kinase activity. The kinase domain of the ALK protein crystallizes as a monomer, and BNP7787-derived mesna-cysteine adducts were observed at Cys 1235 and Cys 1156. The BNP7787-derived mesna adduct at Cys 1156 is located in close proximity to the active site and results in substantial disorder of the P-loop and activation loop (A-loop). Comparison with the P-loop of apo-ALK suggests that the BNP7787-derived mesna adduct at Cys 1156 interferes with the positioning of Phe 1127 into a small pocket now occupied by mesna, resulting in a destabilization of the loop's binding orientation. Additionally, in vitro kinase activity assays indicate that BNP7787 inhibits ALK catalytic activity and potentiates the activity of the ALK-targeted drug crizotinib.

Mitochondrial Dysfunction Confers Albumin-induced NLRP3 Inflammasome Activation and Renal Tubular Injury

Proteinuria is involved in the development of tubular lesions and in the progressive loss of renal function in chronic kidney diseases via uncertain mechanisms. Growing evidence suggests a pathogenic role of mitochondrial dysfunction in chronic kidney diseases. Therefore, the present study aimed to define the roles of mitochondria in proteinuria-induced renal tubular injury and their underlying mechanisms. Using the albumin-overload mouse model, we observed severe tubular structure damage and striking tubular cell apoptosis. Furthermore, tubular epithelial cells displayed a loss of E-cadherin expression and gained expression of α-smooth muscle actin and vimentin, indicating a cellular phenotypic alteration. Strikingly, these albumin overload-induced abnormalities were robustly blocked by a mitochondrial SOD2 mimic, Mn(III) tetrakis (4-benzoic acid)porphyrin chloride (MnTBAP). In agreement with these results, we observed a marked change in mitochondrial morphology accompanied by mitochondrial cytochrome c release and a copy number reduction of mitochondrial DNA. These alterations were largely reversed by MnTBAP, suggesting a key role for mitochondria-derived oxidative stress in mediating the albumin effect on mitochondrial dysfunction and subsequent tubular injury. Moreover, the NOD-like receptor family, pyrin domain-containing 3 (NLRP3)/caspase-1/cytokine cascade was activated in the kidney by albumin overload and was entirely abolished by MnTBAP. In albumin-treated mouse proximal tubular cells, albumin directly induced ROS production, mitochondrial dysfunction, NLRP3/caspase-1/cytokine cascade activation, cell apoptosis, and cellular phenotypic transition. Similar to our in vivo results, treatment with either MnTBAP or cyclosporin A, a mitochondrial permeability transition pore inhibitor, remarkably attenuated these abnormalities in cells. Taken together, these novel findings demonstrate a potential role for the mitochondrial dysfunction/NLRP3 inflammasome axis in the pathogenesis of proteinuria-induced renal tubular injury.

Multidrug Resistance-associated Protein 2 is Involved in the Efflux of Aconitum Alkaloids Determined by MRP2-MDCKII Cells

Aconitum alkaloids mainly contain highly toxic aconitine (AC), mesaconitine (MA), and hypaconitine (HA) and less toxic benzoylaconine (BAC), benzoylmesaconine (BMA), benzoylhypaconine (BHA), aconine, mesaconine, and hypaconine. The efflux transporters including P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and multidrug resistance-associated protein 2 (MRP2) can efflux toxicants to prevent poisoning. Our previous publication has proved that P-gp and BCRP contributed to the efflux of AC, MA and HA, which is demonstrated in the human colonic adenocarcinoma cell lines (Caco-2 cells), Mardin-Darby canine kidney cell lines transfected with MDR1 or BCRP (MDR1-MDCKII and BCRP-MDCKII cells). However, the role of MRP2 remains uncertain.

Novel Implantable Composite Biomaterial by Fibrin Glue and Amniotic Membrane for Ocular Surface Reconstruction

Amniotic membrane transplantation (AMT) is considered a substantial treatment option in the management of ocular surface disorders. However, several inherent drawbacks still remain. The present study devised a novel implantable composite biomaterial of fibrin glue-double layer Amniotic membrane (AM) and evaluated the biomechanical properties and effects on corneal surface reconstruction in alkali-burned rabbit model. Biomechanic parameters were calculated by an electronic universal testing machine. Corneal alkali burning was done in the right eyes of thirty rabbits, which were randomized into three groups of ten animals each. The eyes in group 1 underwent fibrin glue-double layer AMT, the eyes in group 2 underwent ordinary single layer AMT, and the eyes in group 3 (control group) did not undergo any surgical procedure. Healing of corneal epithelial defect, extent of corneal vascularization and corneal clarity were assessed and compared at two time points. One month after surgery, animals were killed and the eyes were processed for histopathology. The fibrin glue-double layer AM composites had more ideal biomechanical properties. In fibrin glue-double layer AM group, the rate of epithelial healing, vascularization inhibition and corneal clarity was significantly better than the other two groups. Novel fibrin glue-double layer AMT with corneal alkali burns is more effective and useful for ocular surface reconstruction and has great potential applications.

Restoring SIRT6 Expression in Hutchinson-Gilford Progeria Syndrome Cells Impedes Premature Senescence and Formation of Dysmorphic Nuclei

Mice overexpressing SIRT6 live longer than wild-type mice while SIRT6 knockout mice exhibit similar degenerative phenotypes as individuals with Hutchinson-Gilford progeria syndrome (HGPS). Thus, we sought to test whether levels of SIRT6 are reduced in cells from individuals with HGPS and whether restored SIRT6 expression may impede premature aging phenotypes.

Complete Blood Count Reference Intervals for Healthy Han Chinese Adults

Complete blood count (CBC) reference intervals are important to diagnose diseases, screen blood donors, and assess overall health. However, current reference intervals established by older instruments and technologies and those from American and European populations are not suitable for Chinese samples due to ethnic, dietary, and lifestyle differences. The aim of this multicenter collaborative study was to establish CBC reference intervals for healthy Han Chinese adults.

In Vitro and in Vivo Ocular Biocompatibility of Electrospun Poly(ɛ-caprolactone) Nanofibers

Biocompatibility is a requirement for the development of nanofibers for ophthalmic applications. In this study, nanofibers were elaborated using poly(ε-caprolactone) via electrospinning. The ocular biocompatibility of this material was investigated. MIO-M1 and ARPE-19 cell cultures were incubated with nanofibers and cellular responses were monitored by viability and morphology. The in vitro biocompatibility revealed that the nanofibers were not cytotoxic to the ocular cells. These cells exposed to the nanofibers proliferated and formed an organized monolayer. ARPE-19 and MIO-M1 cells were capable of expressing GFAP, respectively, demonstrating their functionality. Nanofibers were inserted into the vitreous cavity of the rat's eye for 10days and the in vivo biocompatibility was investigated using Optical Coherence Tomography (OCT), histology and measuring the expression of pro-inflammatory genes (IL-1β, TNF-α, VEGF and iNOS) (real-time PCR). The OCT and the histological analyzes exhibited the preserved architecture of the tissues of the eye. The biomaterial did not elicit an inflammatory reaction and pro-inflammatory cytokines were not expressed by the retinal cells, and the other posterior tissues of the eye. Results from the biocompatibility studies indicated that the nanofibers exhibited a high degree of cellular biocompatibility and short-term intraocular tolerance, indicating that they might be applied as drug carrier for ophthalmic use.

Aberrant Expression of the CHFR Prophase Checkpoint Gene in Human B-cell Non-Hodgkin Lymphoma

Checkpoint with FHA and Ring Finger (CHFR) is a checkpoint protein that reportedly initiates a cell cycle delay in response to microtubule stress during prophase in mitosis, which has become an interesting target for understanding cancer pathogenesis. Recently, aberrant methylation of the CHFR gene associated with gene silencing has been reported in several cancers. In the present study, we examined the expression of CHFR in B-cell non-Hodgkin lymphoma (B-NHL) in vitro and in vivo. Our results showed that the expression level of CHFR mRNA and protein was reduced in B-NHL tissue samples and B cell lines. Furthermore, CHFR methylation was detected in 39 of 122 B-NHL patients, which was not found in noncancerous reactive hyperplasia of lymph node (RH) tissues. CHFR methylation correlated with the reduced expression of CHFR, high International Prognostic Index (IPI) scores and later pathologic Ann Arbor stages of B-NHL. Treatment with demethylation reagent, 5-Aza-dC, could eliminate the hypermethylation of CHFR, enhance CHFR expression and cell apoptosis and inhibit the cell proliferation of Raji cells, which could be induced by high expression of CHFR in Raji cells. Our results indicated that aberrant methylation of CHFR may be associated with the pathogenesis, progression for B-NHL, which might be a novel molecular marker as prognosis and treatment for B-NHL.

(2R,3S)-Pinobanksin-3-cinnamate Improves Cognition and Reduces Oxidative Stress in Rats with Vascular Dementia

This study investigated the neuroprotective effects of (2R,3S)-pinobanksin-3-cinnamate (PNC) in rats with occlusion-damaged bilateral common carotid arteries. Administration with PNC (5 and 10 mg/kg/day) for 5 weeks significantly improved the behavioral performance of rats with vascular dementia, as showed in the Morris water maze test by shortening the escape latency and latency of crossing, completing more platform crossings, as well as spending more time in the target zone. Further evaluations found that PNC could markedly decrease malondialdehyde levels, enhance superoxide dismutase activity and glutathione levels, and decrease the release of cytochrome c as well as the activities of caspases. Moreover, PNC increased Nrf2 and anti-apoptotic bcl-2 protein expression, while Nox1 and pro-apopotic bax protein expression was decreased. PNC may exert its neuroprotective effects through counteracting oxidative stress and has the potential to treat vascular dementia.

Pyrene Fused Perylene Diimides: Synthesis, Characterization and Applications in Organic Field-effect Transistors and Optical Limiting with High Performance

Three pyrene fused PDI derivatives have been obtained, in which totally different properties were observed when adopting different fusing types. For bilaterally benzannulated PDIs, through spin-coating, bottom-contact OFET devices exhibited a p-type mobility up to 1.13 cm(2) V(-1) s(-1), with an on/off ratio of 10(8) in air.

Au Nanoparticles Decorated C60 Nanoparticle-based Label-free Electrochemiluminesence Aptasensor Via a Novel "on-off-on" Switch System

Herein, a label-free electrochemiluminescence (ECL) aptasensor for highly sensitive determination of kanamycin was developed based on a novel "on-off-on" switch system. The first "switch on" state with remarkably high ECL intensity was obtained by the tri-layer composite films modified glassy carbon electrode towards the S2O8(2-)-O2 system. To be specific, the first layer was the Au nanoparticles decorated C60 nanoparticles (abbreviation as Au@nano-C60) as inner-layer which was prepared by the in situ generating of Au nanoparticles onto the surface of bovine serum albumin decorated nano-C60 nanoparticles. Then poly-l-histidine was first selected as a novel coreactant of S2O8(2-)-O2 system and it was adsorbed on the Au@nano-C60 modified electrode as inter-layer. Finally, a self-assembling layer of colloidal Au nanoparticles (AuNPs) was the outer-layer. The three layers were interaction by the Au-N covalent bond which could achieve a desirable initial amplified ECL signal. Successively, the capture probes (CPs) of the aptamer for the target of kanamycin was anchored on the resultant tri-layer composite films modified electrode. Later, the "switch off" state was made by the quenching effect of hemin/G-quadruplex DNAzymes towards S2O8(2-)-O2 system according to the DNA hybridization reaction of an assistant probes (APs, guanine-rich nucleic acid) with CPs which could generate a large amount of hemin/G-quadruplex DNAzymes in the presence of hemin with a simple and label-free process. As expected, the second "switch on" state was the ECL signal recovery when the target of kanamycin was present, it is attributed to that the formation of the aptamer-kanamycin complex makes the quencher of hemin/G-quadruplex DNAzymes release from the sensing interface. With the association of "on-off-on" switch system, a linear response about 9 orders of magnitude for kanamycin detection was obtained from 0.15 nM to 170 mm as well as a detection limit down to 45 pM.

Huaier Cream Protects Against Adriamycin-Induced Nephropathy by Restoring Mitochondrial Function Via PGC-1α Upregulation

The mechanism by which Huaier, a Chinese traditional medicine, protects podocytes remains unclear. We designed the present study to examine whether mitochondrial function restored by PGC-1α serves as the major target of Huaier cream in protecting ADR nephropathy. After ADR administration, the podocytes exhibited remarkable cell injury and mitochondrial dysfunction. Additionally, ADR also reduced PGC-1α both in vivo and in vitro. Following the Huaier treatment, the notable downregulation of PGC-1α and its downstream molecule mitochondrial transcription factor A (TFAM) were almost entirely blocked. Correspondingly, Huaier markedly ameliorated ADR-induced podocyte injury and mitochondrial dysfunction in both rat kidneys and incubated cells as it inhibited the decrease of nephrin and podocin expression, mtDNA copy number, MMP, and ATP content. Transmission electron microscopy result also showed that Huaier protected mitochondria against ADR-induced severe mitophagy and abnormal changes of ultrastructural morphology. In conclusion, Huaier can protect podocytes against ADR-induced cytotoxicity possibly by reversing the dysfunction of mitochondria via PGC-1α overexpression, which may be a novel therapeutic drug target in glomerular diseases.

Clonogenically Culturing and Expanding CD34+ Liver Cancer Stem Cells in Vitro

A large number of cancer stem cells (CSC) have been isolated and identified; however, none has been cultured in an unlimited manner in vitro without losing tumorigenicity and multipotency. Here we successfully clonogenically cultured a newly identified CD34+ liver CSC (LCSC) on feeder cells up to 22 passages (to date) without losing CSC property. Cloned CD34+ LCSC formed a round, packed morphology, and it also could be cryopreserved and re-cultured. Stem cell markers, CD34, CD117 and SOX2; normal liver stem cell markers, alpha fetoprotein, CK19, CK18, and OV6; putative CSC markers, CD44, CD133, EpCAM, and CD90; as well as CD31, were expressed in cloned CD34+ LCSC. SOX2 was the major factor in maintaining this LCSC before colonization, and interestingly, OCT4, SOX2, NAONG, Klf4, c-Myc and Lin28 were up-regulated in association with symmetric self-renewal for colony growth of CD34+ LCSC on feeder cells. Gene expression patterns of in vitro differentiation were consistent with our in vivo finding, furthermore, the tumorigenecity of cloned CD34+ LCSC was not different from uncloned CD34+ LCSC sorted from parental PLC. These results show that our cloned CD34+ LCSC maintained CSC property including self-renewal, bipotency, and tumorigenicity after long-term culture, demonstrating that this LCSC can be cultured in an unlimited manner in vitro. Thus, establishing pure population of CSCs isolated from the patients will provide an opportunity to explore the mechanisms of tumorigenesis and cancer development, and to identify unique biomarkers presenting potential indicators of drug efficacy against CSCs for establishment of novel strategy for cancer therapy.

Delay-induced Instability in a Nutrient-phytoplankton System with Flow

In this paper, a nutrient-phytoplankton system described by a couple of advection-diffusion-reaction equations with delay was studied analytically and numerically. The aim of this research was to provide an understanding of the impact of delay on instability. Significantly, delay cannot only induce instability, but can also promote the formation of spatial pattern via a Turing-like instability. In addition, the theoretical analysis indicates that the flow (advection term) may lead to instability when the delay term exists. By comparison, diffusion cannot result in Turing instability when flow does not exist. Results of numerical simulation were consistent with the analytical results.

A New CRB1 Rat Mutation Links Müller Glial Cells to Retinal Telangiectasia

We have identified and characterized a spontaneous Brown Norway from Janvier rat strain (BN-J) presenting a progressive retinal degeneration associated with early retinal telangiectasia, neuronal alterations, and loss of retinal Müller glial cells resembling human macular telangiectasia type 2 (MacTel 2), which is a retinal disease of unknown cause. Genetic analyses showed that the BN-J phenotype results from an autosomal recessive indel novel mutation in the Crb1 gene, causing dislocalization of the protein from the retinal Müller glia (RMG)/photoreceptor cell junction. The transcriptomic analyses of primary RMG cultures allowed identification of the dysregulated pathways in BN-J rats compared with wild-type BN rats. Among those pathways, TGF-β and Kit Receptor Signaling, MAPK Cascade, Growth Factors and Inflammatory Pathways, G-Protein Signaling Pathways, Regulation of Actin Cytoskeleton, and Cardiovascular Signaling were found. Potential molecular targets linking RMG/photoreceptor interaction with the development of retinal telangiectasia are identified. This model can help us to better understand the physiopathologic mechanisms of MacTel 2 and other retinal diseases associated with telangiectasia.

Experimental Computed Tomography-guided Vena Cava Puncture in Pigs for Percutaneous Brachytherapy of Middle Mediastinal Lymph Node Metastases

Percutaneous brachytherapy is a valuable method for the treatment of lung cancer and mediastinal lymph nodes metastasis. However, in some of the metastatic lymph nodes in the middle mediastinum, the percutaneous approach cannot be used safely due to possible damage to surrounding anatomical structures. We established an animal model (group of 12 pigs) to assess the safety and feasibility of computed tomography (CT)-guided vena cava puncture.

Molecular Insights into Land Snail Neuropeptides Through Transcriptome and Comparative Gene Analysis

Snails belong to the molluscan class Gastropoda, which inhabit land, freshwater and marine environments. Several land snail species, including Theba pisana, are crop pests of major concern, causing extensive damage to agriculture and horticulture. A deeper understanding of their molecular biology is necessary in order to develop methods to manipulate land snail populations.

Electrochemiluminescence-based Detection Method of Lead(ii) Ion Via Dual Enhancement of Intermolecular and Intramolecular Co-reaction

A novel analytical method to design a highly selective and sensitive detection technique for lead(ii) ions (Pb(2+)) detection was developed based on an electrochemiluminescence (ECL) sensor, taking advantage of the high specificity of the aptamer for Pb(2+) and the use of both intermolecular and intramolecular co-reaction to achieve signal enhancement. For sensing interface construction, l-cysteine (Cys) and gold nanostructured layers were electrodeposited on the electrode surface successively, which afforded a large surface area to anchor massive thiol-terminated auxiliary probes (APs) via a thiol-Au interaction. Then, a DNA duplex was generated based on the hybridization of the APs with capture probes (CPs, Pb(2+) specific aptamers). In the presence of Pb(2+), Pb(2+)-induced aptamers were released from the DNA duplex via the formation of a Pb(2+)-stabilized G-quadruplex, accompanied by leaving the single CPs on the sensing interface. Herein, the ruthenium(ii) complexes with functional groups of -COOH (Ru-COOH) were covalently bonded on the polyamidoamine dendrimers with amine end groups (PAMAM), which were capped by the high-index-faceted Au nanoparticles (HIFAuNPs) to obtain the ECL signal labels of Ru-PAMAM-HIFAuNPs. Then, the detection probes (DPs) of amino-terminated Pb(2+) specific aptamers were tagged with the Ru-PAMAM-HIFAuNPs. It was demonstrated that the covalent bonding of PAMAM and Ru-COOH could generate a self-enhanced ECL luminophore by an intramolecular co-reaction and the use of a Cys layer modified electrode could enhance the ECL by the intermolecular co-reaction of Cys and Ru-COOH, which lead to a significant enhancement of the ECL response. Based on this analytical method, the ECL signal increased with Pb(2+) concentration which presented a linear relationship in the range 1.0 × 10(-13)-1.0 × 10(-7) M with the detection limit of 4.0 × 10(-14) M. The proposed approach was also successfully utilized for the determination of Pb(2+) in soil samples.

Hippocampal Volume Reduction in Female but Not Male Recent Abstinent Methamphetamine Users

Growing evidence suggests abnormalities in brain morphology including hippocampal structure in patients with methamphetamine (MA) dependence. This study was performed to examine hippocampal volume in abstinent MA users, and to further explore its relationship with cognitive function. 30 abstinent MA users (20 males and 10 females) with average 5.52 months of duration of abstinence and 29 healthy controls (19 males and 10 females) age 18 to 45 years old were recruited for clinical assessment and imaging scan. FreeSurfer was used to segment the hippocampus bilaterally, and hippocampal volumes were extracted for group and gender comparisons. Cognitive function was measured using the CogState Battery Chinese language version (CSB-C).Analysis of covariance (ANCOVA) controlling for education showed a significant group by gender interaction for the right hippocampal relative volume adjusted for total brain size (p=0.020); there was a significant difference between male controls and female controls (p<0.001), but such a difference didn't exist between male patients and female patients (p=0.203). No significant correlations were found between hippocampal volume and cognitive measures. There seems to be a gender difference in how MA affects hippocampal volume in abstinent MA users. Hippocampus might be an important treatment target for cognitive improvement and functional recovery in this patient population, especially in females.

UPLC-Q-TOF/MS-based Screening and Identification of the Main Flavonoids and Their Metabolites in Rat Bile, Urine and Faeces After Oral Administration of Scutellaria Baicalensis Extract

Traditional Chinese Medicines (TCMs) are increasingly used in combination with Western medicine. Scutellaria baicalensis Georgi (Lamiaceae) is a widely used TCM in treating various diseases. However, the in vivo metabolism of its main bioactive flavonoids, baicalin, baicalein, wogonoside and wogonin, need further study.

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