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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).

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.

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