A New Kinetic Method for Quantification Phenoxyl Free Radicals

Talanta. Sep, 2006  |  Pubmed ID: 18970770

In this work, a new kinetic method was proposed for quantification phenoxyl radicals generated in enzyme reaction. Instead of direct detecting the spectral signals of phenoxyl radicals, a molecular probe, the reduced form of nicotinamide adenine dinucleotide (NADH), was employed to indicate the formation of phenoxyl free radicals. It was found that the reactions of NADH and phenoxyl radicals are very fast, but can be followed by using stopped-flow fast scanning spectrophotometric technique. The initial rate of accelerated-oxidation of NADH represents the reactivity of phenoxyl free radical, which is proportional in a certain range to the initial concentration of the parent chlorophenols of the radicals. With this method, the phenoxyl radicals generated in oxidation reaction of chlorophenols (2-CP; 4-CP; 2,4-DCP; 2,4,6-TCP and 2,3,4,6-Tetra-CP) with hydrogen peroxide, catalyzed by horseradish peroxidase, were investigated. The method is highly sensitive. Phenoxyl radicals generated from as low as 1x10(-8)M 2,4-DCP, for example, can be readily detected with the proposed method. The results show that the reactivity of various phenoxyl radicals are in the following order: 2,4-DCP>4-CP>2-CP>2,4,6-TCP>2,3,4,6-Tetra-CP. A mechanism is proposed to explain the possible pathway of the probe reaction. The feasibility of this method was assessed by the determination of enzymatic generation of phenoxyl radicals in lake water samples.

Expression of Vascular Endothelial Growth Factor in Human Meningiomas and Peritumoral Brain Areas

Annals of Clinical and Laboratory Science. 2008  |  Pubmed ID: 18988927

Vascular endothelial growth factor (VEGF) is a regulator of angiogenesis, vasculogenesis, and vascular permeability. Recent reports suggest that VEGF may play a critical role in formation of peritumoral brain edema (PTBE) associated with meningiomas. While VEGF expression has been shown in meningiomas, studies have not focused on VEGF in the adjacent peritumoral brain regions. The present study examined the protein and gene expression of VEGF in human meningiomas and peritumoral brain areas. Biopsies were obtained from 37 patients. Immunohistochemical staining and immunoblotting were performed to detect the expression of VEGF protein. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to analyze the presence and quantity of VEGF mRNA. The extent of PTBE was estimated as an edema index (EI) based on preoperative magnetic resonance imaging. In meningiomas, western blot and RT-PCR results were congruent and the expression of both protein and mRNA had a significant correlation with EI. However, in peritumoral areas, western blot results were not consistent with the RT-PCR results. Protein results showed high correlation with EI, but mRNA was almost undetectable. In VEGF-positive cases, a decreasing gradient of VEGF protein expression was observed with increasing distance from tumors. These data suggest that peritumoral tissue does not produce VEGF and that VEGF protein levels in peritumoral tissues have a high correlation with EI. We conclude that VEGF macromolecules are secreted by the tumor tissue and enter peritumoral normal brain tissue to induce edema.

CuS Nanotubes for Ultrasensitive Nonenzymatic Glucose Sensors

Chemical Communications (Cambridge, England). Dec, 2008  |  Pubmed ID: 19030547

CuS nanotubes made up of nanoparticles were successfully prepared in large quantities in an O/W microemulsion system under low temperature; the as-prepared CuS nanotube modified electrode was used as an enzyme-free glucose sensor.

[Recombinant Envelope Glycoprotein Domain III of Dengue Virus Inhibit Virus Infection]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Jun, 2008  |  Pubmed ID: 19031695

To observe the ability of dengue virus recombinant envelope protein domain expressed in E. coli to inhibit virus infection and induce the neutralizing antibody.

Open Waveguide Cavity Using a Negative Index Medium

Optics Letters. Dec, 2008  |  Pubmed ID: 19037433

An open waveguide cavity formed by a pair of planar waveguides, in which one guiding layer is a negative index medium and the other is a positive index medium, is theoretically demonstrated. For such a waveguide cavity the resonant frequency is independent of the total length of the waveguide system. With the coupled mode theory it is shown that energy flow circulation can be established through the special coupling between the waveguides at the resonant frequency, and thus the wave fields are localized. This phenomenon is further verified numerically with the finite-difference time-domain method. The quality factor of the open waveguide cavity is also discussed.

Synthesis and Binding Site Characteristics of 2,4,6-trichlorophenol-imprinted Polymers

Analytical and Bioanalytical Chemistry. Dec, 2008  |  Pubmed ID: 18820903

2,4,6-Trichlorophenol (2,4,6-TCP)-imprinted micro- and submicrospheres prepared by precipitation polymerization were compared with templated materials obtained by conventional bulk polymerization. The influence of the type and amount of functional monomer, the type and amount of cross-linker, polymerization temperature, porogen, and the ratio of template molecule and functional monomer to cross-linker on the size of the obtained particles were investigated. UV-Vis spectrophotometer experiments revealed that the microsphere polymers provided higher affinity to the template in contrast to imprinted polymers prepared by bulk polymerization. The binding properties of the microspheres, including binding isotherms and affinity distribution, were studied via Freundlich isotherm affinity distribution (FIAD) analysis. The obtained results indicated that microspheres prepared by precipitation polymerization provided superior rebinding properties during equilibrium binding in contrast to bulk polymers and submicrosphere polymers. Moreover, release experiments showed that 80% of rebound 2,4,6-TCP was released from the imprinted microspheres within the first 2 h, while more intimately bound 2,4,6-TCP molecules were released in the following 40 h. The morphologies and porosities of the resulting imprinted materials were characterized by scanning electron microscopy (SEM) and Brunauer-Emmett-Teller (BET) analysis, respectively. The microsphere polymers exhibited a regular spherical shape with a high degree of monodispersity to the corresponding bulk polymers. Furthermore, the micro- and submicrospheres were characterized by narrow distribution of pores in contrast to a heterogeneity index of m = 0.6647 for the microsphere imprinted polymer.

[Inhibition of HSP70-2 Expression by RNA Interference Induces Apoptosis of Human Hepatocellular Carcinoma Cells]

Zhonghua Gan Zang Bing Za Zhi = Zhonghua Ganzangbing Zazhi = Chinese Journal of Hepatology. Sep, 2008  |  Pubmed ID: 18822209

To determine the effect of short hairpin RNA targeting HSP70-2 on the growth and apoptosis of hepatocellular carcinoma (HCC) cells, and to elucidate its possible mechanisms in mitochondria apoptotic pathway.

The Neuronal Activity of Thalamic Parafascicular Nucleus is Conversely Regulated by Nigrostriatal Pathway and Pedunculopontine Nucleus in the Rat

Brain Research. Nov, 2008  |  Pubmed ID: 18823953

The aim of the present study was to investigate changes in the firing activity of thalamic parafascicular nucleus (PF) neurons at different time periods after 6-hydroxydopamine (6-OHDA) lesions of the substantia nigra pars compacta (SNc) and the role of the pedunculopontine nucleus (PPN) in these changes. In normal rats, the firing rate of PF neurons was 3.66+/-0.37 spikes/s. In rats with 6-OHDA lesions of the SNc, the firing rate of PF neurons slightly decreased to 3.19+/-0.35 spikes/s during the third week compared to normal rats, unexpectedly, as moving on to fifth week, the firing rate increased significantly to 4.82+/-0.31 spikes/s. In rats with ibotenic acid lesions of the PPN, the firing rate decreased significantly to 1.98+/-0.19 spikes/s compared to normal rats. When the SNc and PPN were double lesioned, the firing rate of PF neurons decreased significantly to 2.36+/-0.23 spikes/s during the third week and 2.16+/-0.16 spikes/s during the fifth week post-lesions. The separate lesions of the PPN, SNc, and double lesion of both in the rats did not change the firing pattern of PF neurons compared to normal rats. These findings demonstrate that PF neurons are hyperactive in the 6-OHDA-lesioned rats suggesting the implication of this nucleus in the pathophysiology of parkinsonism. Furthermore, the fact that the PPN lesions induced a decrease in the firing rate of PF neurons in normal and SNc-lesioned rats suggests that the PF is under major control of the PPN.

The Firing Activity of Presumed Cholinergic and Non-cholinergic Neurons of the Pedunculopontine Nucleus in 6-hydroxydopamine-lesioned Rats: an in Vivo Electrophysiological Study

Brain Research. Dec, 2008  |  Pubmed ID: 18824158

Several studies have shown that the neuronal activity of the pedunculopontine nucleus is increased in Parkinson's disease. In the present study, the changes were examined in the firing rate and firing pattern of presumed cholinergic and non-cholinergic neurons in the pedunculopontine nucleus of 6-hydroxydopamine-lesioned rats by using extracellular recording. In the lesioned rats, the mean firing rate of both presumed cholinergic and non-cholinergic neurons in the pedunculopontine nucleus increased significantly compared to normal rats. With regard to firing pattern, the majority of presumed cholinergic and non-cholinergic neurons fired regularly in normal rats. After substantia nigra pars compacta-lesion, the percentage of presumed non-cholinergic neurons exhibiting irregular pattern increased significantly compared to normal rats, while having no significant change in the firing pattern of presumed cholinergic neurons. Collectively, these results indicate that the presumed cholinergic and non-cholinergic neurons in the pedunculopontine nucleus are overactive in 6-hydroxydopamine-lesioned rats, particularly, presumed non-cholinergic neuron firing is more irregular, which suggests that the firing activity of presumed cholinergic and non-cholinergic neurons is affected by the different afferents from the basal ganglia and related structures.

S-nitrosylated S100A8: Novel Anti-inflammatory Properties

Journal of Immunology (Baltimore, Md. : 1950). Oct, 2008  |  Pubmed ID: 18832721

S100A8 and S100A9, highly expressed by neutrophils, activated macrophages, and microvascular endothelial cells, are secreted during inflammatory processes. Our earlier studies showed S100A8 to be an avid scavenger of oxidants, and, together with its dependence on IL-10 for expression in macrophages, we postulated that this protein has a protective role. S-nitrosylation is an important posttranslational modification that regulates NO transport, cell signaling, and homeostasis. Relatively few proteins are targets of S-nitrosylation. To date, no inflammation-associated proteins with NO-shuttling capacity have been identified. We used HPLC and mass spectrometry to show that S100A8 and S100A9 were readily S-nitrosylated by NO donors. S-nitrosylated S100A8 (S100A8-SNO) was the preferred nitrosylated product. No S-nitrosylation occurred when the single Cys residue in S100A8 was mutated to Ala. S100A8-SNO in human neutrophils treated with NO donors was confirmed by the biotin switch assay. The stable adduct transnitrosylated hemoglobin, indicating a role in NO transport. S100A8-SNO suppressed mast cell activation by compound 48/80; intravital microscopy was used to demonstrate suppression of leukocyte adhesion and extravasation triggered by compound 48/80 in the rat mesenteric microcirculation. Although S100A8 is induced in macrophages by LPS or IFN-gamma, the combination, which activates inducible NO synthase, did not induce S100A8. Thus, the antimicrobial functions of NO generated under these circumstances would not be compromised by S100A8. Our results suggest that S100A8-SNO may regulate leukocyte-endothelial cell interactions in the microcirculation, and suppression of mast cell-mediated inflammation represents an additional anti-inflammatory property for S100A8.

Determination of Estrogens and Their Metabolites in Water Using C30 SPE-LC-MS

Journal of Separation Science. Oct, 2008  |  Pubmed ID: 18837473

Triacontyl bonded silica (C(30)) material was applied as solid-phase extraction (SPE) sorbent and an SPE-LC-MS method was established for the determination of eight estrogens and their metabolites in water samples. Compared to commercial C(18) SPE cartridge, the performance of C(30) was evaluated in various important SPE conditions, such as sorbent mass, elution solvent and its volume, loading flow rate, and sample loading volume. The results showed a superior performance of C(30)-C(18) by the shorter treatment time and fewer required elution solvent. In the optimum conditions, the results showed good recoveries (80.5-109.4%), excellent linear relationships (0.02-1 ng/mL, except 2-MeO-E(2)), high precisions (lower than 10.0% RSD for both low and high spiked concentration), and low LODs (1-16 ng/L). Method validation using C(30) packed cartridge was also testified with spiked real water samples, including tap water and river water. Satisfy results demonstrated the feasibility of C(30) SPE to the analysis for real environmental waters.

Cephalosol: an Antimicrobial Metabolite with an Unprecedented Skeleton from Endophytic Cephalosporium Acremonium IFB-E007

Chemistry (Weinheim an Der Bergstrasse, Germany). 2008  |  Pubmed ID: 18850610

Cephalosol (1), a potent antimicrobial secondary metabolite with a new carbon skeleton, was characterized from the culture of Cephalosporium acremonium IFB-E007 that used to reside as an endophyte in Trachelospermum jasminoides (Apocynaceae). Its structure and absolute configuration were unambiguously determined by spectroscopic and computational approaches.

Effect of Tang No.1 Granule (1) in Treating Patients with Impaired Glucose Tolerance

Chinese Journal of Integrative Medicine. Dec, 2008  |  Pubmed ID: 19082803

To observe the therapeutic effect of Tang No.1 granule (1, T1G) in treating patients with impaired glucose tolerance (IGT).

[Transperitoneal Laparoscopic Adrenalectomy: Surgical Approach and Outcome]

Zhonghua Wai Ke Za Zhi [Chinese Journal of Surgery]. Dec, 2008  |  Pubmed ID: 19134375

To investigate the efficiency and safety of transperitoneal laparoscopic adrenalectomy for the treatment of adrenal tumors and to describe surgical technique and management of intraoperative complications.

[Chiral Separation of Racemic Epoxiconazole on Cellulose Tris(3,5-dimethylphenylcarbamate) Chiral Stationary Phase Using High Performance Liquid Chromatography]

Se Pu = Chinese Journal of Chromatography / Zhongguo Hua Xue Hui. Sep, 2008  |  Pubmed ID: 19160767

The two pairs of enantiomers of epoxiconazole were resolved using normal phase, reversed-phase and polar organic solvent high performance liquid chromatography on cellulose tris(3,5-dimethylphenylcarbamate) (CDMPC) chiral stationary phase, separately. The effects of different mobile phase compositions on the retention factor, separation factor and resolution in the chiral separation of epoxiconazole were investigated. It can be baseline separated on a Chiralcel OD-H column packed with CDMPC chiral stationary phase when methanol-water (80: 20, v/v) was used as the mobile phase, the resolutions of the two pairs of enantiomers were 1.64 and 6.50, respectively.

[Follow-up Study of Traumatic Macular Hole]

[Zhonghua Yan Ke Za Zhi] Chinese Journal of Ophthalmology. Sep, 2008  |  Pubmed ID: 19175156

To investigate the natural course and visual outcomes of traumatic macular hole.

Disruption of Nrf2 Enhances Upregulation of Nuclear Factor-kappaB Activity, Proinflammatory Cytokines, and Intercellular Adhesion Molecule-1 in the Brain After Traumatic Brain Injury

Mediators of Inflammation. 2008  |  Pubmed ID: 19190763

Inflammatory response plays an important role in the pathogenesis of secondary brain injury after traumatic brain injury (TBI). Nuclear factor erythroid 2-related factor 2 (Nrf2) is a key transcription factor that plays a crucial role in cytoprotection against inflammation. The present study investigated the role of Nrf2 in the cerebral upregulation of NF-kappaB activity, proinflammatory cytokine, and ICAM-1 after TBI. Wild-type Nrf2 (+/+) and Nrf2 (-/-)-deficient mice were subjected to a moderately severe weight-drop impact head injury. Electrophoretic mobility shift assays (EMSAs) were performed to analyze the activation of nuclear factor kappa B (NF-kappaB). Enzyme-linked immunosorbent assays were performed to quantify the production of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6). Immunohistochemistry staining experiments were performed to detect the expression of intercellular adhesion molecule-1 (ICAM-1). Nrf2 (-/-) mice were shown to have more NF-kappaB activation, inflammatory cytokines TNF-alpha, IL-1beta and IL-6 production, and ICAM-1 expression in brain after TBI compared with their wild-type Nrf2 (+/+) counterparts. The results suggest that Nrf2 plays an important protective role in limiting the cerebral upregulation of NF-kappaB activity, proinflammatory cytokine, and ICAM-1 after TBI.

Pravastatin Prevents Aortic Atherosclerosis Via Modulation of Signal Transduction and Activation of Transcription 3 (STAT3) to Attenuate Interleukin-6 (IL-6) Action in ApoE Knockout Mice

International Journal of Molecular Sciences. Nov, 2008  |  Pubmed ID: 19330073

The purpose of this study was to determine whether pravastatin's prevention of aortic atherosclerosis via attenuation of IL-6 action depends on modulation of STAT3 activity. Male apoE knockout (apoE-/-) mice fed on a diet containing 1.25% cholesterol (wt/wt) were divided into pravastatin group provided with pravastatin (80 mg kg(-1) per day) and atherosclerosis group. After eight weeks, pravastatin significantly prevented atherosclerotic lesion and reduced levels of IL-6 in serum and lesion, and significantly decreased expressions of phosphorylated STAT3 (pSTAT3) and increased suppressor of cytokine signaling 3 (SOCS3) expressions in lesions. Our results suggested that pravastatin's aortic atherosclerosis preventing action via attenuation of IL-6 action may partially depend on modulation of STAT3 activity.

[Hypoxia Induces Heat Shock Protein HSP70-2 Expression in a HIF-1 Dependent Manner]

Zhonghua Gan Zang Bing Za Zhi = Zhonghua Ganzangbing Zazhi = Chinese Journal of Hepatology. Mar, 2009  |  Pubmed ID: 19335985

To investigate role of hypoxia inducible factor 1 (HIF-1) in the transcriptional activation of heat shock protein 70-2 (HSP70-2) in hepatocellular carcinoma (HCC) cells under hypoxic conditions.

Transcription Factor Nrf2 Plays a Pivotal Role in Protection Against Traumatic Brain Injury-induced Acute Intestinal Mucosal Injury in Mice

The Journal of Surgical Research. Dec, 2009  |  Pubmed ID: 19394962

Traumatic brain injury (TBI) can induce an acute intestinal mucosal injury. Nuclear factor erythroid 2-related factor 2 (Nrf2) has a unique role in many physiological stress processes, but its contribution to intestinal mucosal injury after TBI remains to be determined.

Molecularly Imprinted Solid-phase Extraction for the Selective Determination of 17beta-estradiol in Fishery Samples with High Performance Liquid Chromatography

Talanta. Apr, 2009  |  Pubmed ID: 19203606

A molecularly imprinted polymer (MIP) has been synthesized by a thermo-polymerization method using methacrylic acid (MAA) as functional monomer, ethylene glycol dimethacrylate (EGDMA) as cross-linker, acetonitrile as porogenic solvent, and 17beta-estradiol as template. The MIP showed obvious affinity for 17beta-estradiol in acetonitrile solution, which was confirmed by absorption experiments. After optimization of molecularly imprinted solid-phase extraction (MISPE) conditions, three structurally related estrogenic compounds (17beta-estradiol, estriol, and diethylstilbestrol) were used to evaluate the selectivity of the MIP cartridges. The MIP cartridges exhibited highly selectivity for E(2), the recoveries were 84.8+/-6.53% for MIPs and 19.1+/-1.93% for non-imprinted polymer (NIP) cartridges. The detection and quantification limits correspond to 0.023 and 0.076 mg L(-1). Furthermore, the MISPE methods were used to selectively extract E(2) from fish and prawn tissue prior to HPLC analysis. This MISPE-HPLC procedure could eliminate all matrix interference simultaneously and had good recoveries (78.3-84.5%).

Rapid Glycation with D-ribose Induces Globular Amyloid-like Aggregations of BSA with High Cytotoxicity to SH-SY5Y Cells

BMC Cell Biology. 2009  |  Pubmed ID: 19216769

D-ribose in cells and human serum participates in glycation of proteins resulting in advanced glycation end products (AGEs) that affect cell metabolism and induce cell death. However, the mechanism by which D-ribose-glycated proteins induce cell death is still unclear.

Molecularly Imprinted Solid-phase Extraction Combined with High Performance Liquid Chromatography for Analysis of Phenolic Compounds from Environmental Water Samples

Journal of Hazardous Materials. Aug, 2009  |  Pubmed ID: 19233552

The molecularly imprinted bulk polymer with 2,4,6-trichlorophenol (2,4,6-TCP) as the template molecule and methylacrylic acid (MAA), ethylene glycol dimethacrylate (EGDMA) as functional monomer and the crosslinker, respectively, has been prepared and applied to the molecularly imprinted solid-phase extraction (MISPE) procedure for selective preconcentration of phenolic compounds from environmental water samples. Various parameters affecting the extraction efficiency of the polymer have been evaluated to optimize the selective preconcentration of the phenolic compounds from aqueous samples. The characteristics of the MISPE method were validated by HPLC. The recoveries ranged between 90% and 98% (RSD: 0.9-2.3%, n=3) for tap water, between 85% and 105% (RSD: 2.6-4.9%, n=3) for river water, between 78% and 98% (RSD: 2.6-5.4%, n=3) for sewage water fortified with 0.4 mg L(-1) of phenol, 4-chlorophenol (4-CP), 2,4-dichlorophenol (2,4-DCP), pentachlorophenol (PCP). It was demonstrated that this MISPE-HPLC method could be applied to direct preconcentration and determination of phenolic compounds in environmental water samples.

Addition of an Alginate to a Modified Zeolite Improves Hemostatic Performance in a Swine Model of Lethal Groin Injury

The Journal of Trauma. Mar, 2009  |  Pubmed ID: 19276728

QuikClot is a zeolite-based hemostatic agent that can control severe hemorrhage through adsorption of water in an exothermic reaction. Ion exchanging the calcium ions in zeolite type 5A with cations of a reduced hydration enthalpy can reduce heat generation, but its effect on the hemostatic efficacy is not clear. We developed a new compound zeolite hemostat and tested it against controls in a modified swine model of battlefield injury.

PI3 Kinase/Akt Signaling Mediates Epithelial-mesenchymal Transition in Hypoxic Hepatocellular Carcinoma Cells

Biochemical and Biophysical Research Communications. May, 2009  |  Pubmed ID: 19303863

Hypoxia activates genetic programs that facilitate cell survival; however, in cancer, it may promote invasion and metastasis. Although the exact mechanisms driving hypoxia-induced invasion and metastasis remain elusive, we hypothesized that epithelial-mesenchymal transition (EMT) may play a major role. We investigated this in vitro by treating hepatocellular carcinoma cells under 1.0% O(2). After the hypoxia treatment, the cells exhibited some morphological changes including cell elongation, cytoskeletal rearrangement, and junctional disruption. Moreover, expression of the epithelia-specific marker E-cadherin was decreased and expression of the myofibroblast-specific marker vimentin was detected in the treated cells. Cell migration and ECM gel invasion were increased. These findings were consistent with events observed during EMT. Hypoxia-induced EMT is accompanied by increased phosphorylation, activation of Akt and the downstream signaling. Hypoxia-induced EMT was blocked by PI3K inhibitor LY294002. The results suggest that the PI3K/Akt-dependent signaling pathways serve to regulate hypoxia-induced EMT of hepatocellular carcinoma cells.

Many X-linked MicroRNAs Escape Meiotic Sex Chromosome Inactivation

Nature Genetics. Apr, 2009  |  Pubmed ID: 19305411

Meiotic sex chromosome inactivation (MSCI) during spermatogenesis is characterized by transcriptional silencing of genes on both the X and Y chromosomes in mid-to-late pachytene spermatocytes. MSCI is believed to result from meiotic silencing of unpaired DNA because the X and Y chromosomes remain largely unpaired throughout first meiotic prophase. However, unlike X-chromosome inactivation in female embryonic cells, where 25-30% of X-linked structural genes have been reported to escape inactivation, previous microarray- and RT-PCR-based studies of expression of >364 X-linked mRNA-encoding genes during spermatogenesis have failed to reveal any X-linked gene that escapes the silencing effects of MSCI in primary spermatocytes. Here we show that many X-linked miRNAs are transcribed and processed in pachytene spermatocytes. This unprecedented escape from MSCI by these X-linked miRNAs suggests that they may participate in a critical function at this stage of spermatogenesis, including the possibility that they contribute to the process of MSCI itself, or that they may be essential for post-transcriptional regulation of autosomal mRNAs during the late meiotic and early postmeiotic stages of spermatogenesis.

[An Investigation of Prostate Cancer Knowledge Among Chinese City Men]

Zhonghua Nan Ke Xue = National Journal of Andrology. Feb, 2009  |  Pubmed ID: 19323376

To investigate the knowledge and the attitude toward prostate cancer (PCa) among Chinese city men and raise some suggestions for the improvement of the present state.

Genetic Ablation of Nrf2 Enhances Susceptibility to Acute Lung Injury After Traumatic Brain Injury in Mice

Experimental Biology and Medicine (Maywood, N.J.). Feb, 2009  |  Pubmed ID: 19176347

Previous studies have shown that nuclear factor erythroid 2-related factor 2 (Nrf2) plays a unique role in many physiological stress processes. The present study investigated the role of Nrf2 in the regulation of traumatic brain injury (TBI)-induced acute lung injury (ALI). Wild-type Nrf2 (+/+) and Nrf2 (-/-)-deficient mice were subjected to a moderately severe weight-drop impact head injury. Pulmonary capillary permeability (PCP), wet/dry weight ratio, apoptosis, inflammatory cytokines and antioxidant/detoxifying enzymes were measured at 24 h after TBI. Mice lacking Nrf2 were found to be more susceptible to TBI-induced ALI, as characterized by the higher increase in PCP, wet/dry weight ratio and alveolar cells apoptosis after TBI. This exacerbation of lung injury in Nrf2-deficient mice was associated with increased pulmonary mRNA and protein expression of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and interleukin-6 (IL-6); and with decreased pulmonary mRNA expression and enzymatic activities of antioxidant and detoxifying enzymes including NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S-transferase alpha1 (GST-alpha1)--as compared with their wild-type Nrf2 (+/+) counterparts after TBI. The results of the present study suggest that Nrf2 reduces TBI-induced acute lung injury, possibly by decreasing pulmonary inflammation and inducing antioxidant and detoxifying enzymes.

PP4 and PP2A Regulate Hedgehog Signaling by Controlling Smo and Ci Phosphorylation

Development (Cambridge, England). Jan, 2009  |  Pubmed ID: 19088085

The seven-transmembrane protein Smoothened (Smo) and Zn-finger transcription factor Ci/Gli are crucial components in Hedgehog (Hh) signal transduction that mediates a variety of processes in animal development. In Drosophila, multiple kinases have been identified to regulate Hh signaling by phosphorylating Smo and Ci; however, the phosphatase(s) involved remain obscured. Using an in vivo RNAi screen, we identified PP4 and PP2A as phosphatases that influence Hh signaling by regulating Smo and Ci, respectively. RNAi knockdown of PP4, but not of PP2A, elevates Smo phosphorylation and accumulation, leading to increased Hh signaling activity. Deletion of a PP4-interaction domain (amino acids 626-678) in Smo promotes Smo phosphorylation and signaling activity. We further find that PP4 regulates the Hh-induced Smo cell-surface accumulation. Mechanistically, we show that Hh downregulates Smo-PP4 interaction that is mediated by Cos2. We also provide evidence that PP2A is a Ci phosphatase. Inactivating PP2A regulatory subunit (Wdb) by RNAi or by loss-of-function mutation downregulates, whereas overexpressing regulatory subunit upregulates, the level and thus signaling activity of full-length Ci. Furthermore, we find that Wdb counteracts kinases to prevent Ci phosphorylation. Finally, we have obtained evidence that Wdb attenuates Ci processing probably by dephosphorylating Ci. Taken together, our results suggest that PP4 and PP2A are two phosphatases that act at different positions of the Hh signaling cascade.

Transcriptional Up-regulation of FoxM1 in Response to Hypoxia is Mediated by HIF-1

Journal of Cellular Biochemistry. Feb, 2009  |  Pubmed ID: 19097132

The proliferation-specific Forkhead box M1 (FoxM1) transcription factor is overexpressed in cancer cells and acts as an important regulator of cancer cell growth and survival. Here, we show the molecular mechanisms by which hypoxia regulate FoxM1 expression in cancer cells. When cells were subjected to hypoxia (1% O2), the mRNA and protein levels of FoxM1 had a significant increase in cancer cells (HepG2, MCF-7, and HeLa). Such increase was due to the direct binding of hypoxia-inducible factor 1 (HIF-1) to the HIF-1 binding sites in the FoxM1 promoter. By deletion and mutation assays, we demonstrated that the HIF1-1 and HIF1-3/4 binding sites on the FoxM1 promoter were essential for transcriptional activation of FoxM1 by hypoxia. We also demonstrated that HIF-1alpha directly bound to the promoter of FoxM1 and the binding was specific, as revealed by HIF-1 binding/competition assay and chromatin immunoprecipitation assay. Consequently, the up-regulation of FoxM1 accelerated the growth of hypoxic cancer cells by decreasing nuclear levels of p21 and increasing expression of cyclin B1 and cyclin D1. These findings provide a new insight into how tumor cells overcome hypoxic stress and survive, and also disclose a new regulatory mechanism of FoxM1 expression in tumor cells.

Evolution of Organelle-associated Protein Profiling

Journal of Proteomics. Feb, 2009  |  Pubmed ID: 19110081

Identification of the protein constituents of cell organelles forms the basis for studies to define the roles of specific proteins in organelle structure and functions. Over the past decade, the use of mass spectrometry-based proteomics has dissected various organelles and allowed the association of many novel proteins with particular organelles. This review chronicles the evolution of organelle proteomics technology, and discusses how many limitations, such as organelle heterogeneity and purity, can be avoided with recently developed quantitative profiling approaches. Although many challenges remain, quantitative profiling of organelles holds the promise to begin to address the complex and dynamic shuttling of proteins among organelles that will be critical for application of this advanced technology to disease-based changes in organelle function.

Arthroderma Vanbreuseghemii Infection in Three Family Members with Kerion and Tinea Corporis

Medical Mycology : Official Publication of the International Society for Human and Animal Mycology. 2009  |  Pubmed ID: 19115135

We present a familial infection caused by Arthroderma vanbreuseghemii. The proband is a 4-year-old boy, who had played with rabbits at his rabbit-farm neighbor. He complained of pruritus and pain in his scalp, which displayed redness, alopecia and painful cysts and eventually discharged pus and scabbed. Several erythema on his face and abdomen were also presented. He was diagnosed as having impetigo but antibacterial agents were not effective and his clinical condition did not improve. Several days later, his parents also developed facial erythema and scaling. The development of a kerion in the boy and tinea corporis in his parents were diagnosed based on the positive KOH examination. Morphologic and biochemical characteristics confirmed that their infections were caused by the zoophilic Trichophyton mentagrophytes, while sequencing of the internal transcribed spacer (ITS) 1/4 polymerase chain reaction products, amplified from primary culture isolates, established its Arthroderma vanbreuseghemii lineage. Random amplified polymorphic DNA (RAPD) analysis indicated these isolates might be the same strain and that infection cruciata occurred in this family. Semi-quantitative analysis of these strains indicated multiple and main enzymatic activities of alkaline phosphatase, beta-glucosaccharase. The boy was cured through treatment with itraconazole 100 mg/day orally in combination with topical washes with 2% ketoconazole shampoo, and his parents were successfully treated by topical application of terbinafine cream.

Role of Nrf2 in Protection Against Traumatic Brain Injury in Mice

Journal of Neurotrauma. Jan, 2009  |  Pubmed ID: 19125683

Previous studies have shown that nuclear factor erythroid 2-related factor 2 (Nrf2) plays a unique role in many physiological stress processes. The present study investigated the role of Nrf2 in modulating traumatic brain injury (TBI)-induced secondary brain injury. Wild-type Nrf2 (+/+) and Nrf2 (-/-)-deficient mice were subjected to a moderately severe weight-drop impact head injury. The absence of Nrf2 function in mice resulted in exacerbated brain injury as shown by the increased severity of neurological deficit, apoptosis, and brain edema at 24h after TBI. This exacerbation of brain injury in Nrf2-deficient mice was associated with increased mRNA and protein expression of inflammatory cytokines such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6), and with decreased mRNA expression and enzymatic activity of antioxidant and detoxifying enzymes including NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S-transferase alpha-1 (GST-alpha1), compared with their wild-type counterparts after TBI. In combination, these results suggest that Nrf2 plays an important role in protecting TBI-induced secondary brain injury, possibly by regulating inflammatory cytokines and inducing antioxidant and detoxifying enzymes.

Prequips--an Extensible Software Platform for Integration, Visualization and Analysis of LC-MS/MS Proteomics Data

Bioinformatics (Oxford, England). Mar, 2009  |  Pubmed ID: 19129212

SUMMARY: We describe an integrative software platform, Prequips, for comparative proteomics-based systems biology analysis that: (i) integrates all information generated from mass spectrometry (MS)-based proteomics as well as from basic proteomics data analysis tools, (ii) visualizes such information for various proteomic analyses via graphical interfaces and (iii) links peptide and protein abundances to external tools often used in systems biology studies. AVAILABILITY: http://prequips.sourceforge.net

Sertoli Cell Dicer is Essential for Spermatogenesis in Mice

Developmental Biology. Feb, 2009  |  Pubmed ID: 19071104

Spermatogenesis requires intact, fully competent Sertoli cells. Here, we investigate the functions of Dicer, an RNaseIII endonuclease required for microRNA and small interfering RNA biogenesis, in mouse Sertoli cell function. We show that selective ablation of Dicer in Sertoli cells leads to infertility due to complete absence of spermatozoa and progressive testicular degeneration. The first morphological alterations appear already at postnatal day 5 and correlate with a severe impairment of the prepubertal spermatogenic wave, due to defective Sertoli cell maturation and incapacity to properly support meiosis and spermiogenesis. Importantly, we find several key genes known to be essential for Sertoli cell function to be significantly down-regulated in neonatal testes lacking Dicer in Sertoli cells. Overall, our results reveal novel essential roles played by the Dicer-dependent pathway in mammalian reproductive function, and thus pave the way for new insights into human infertility.

Differential Expression Profiles of MicroRNAs in NIH3T3 Cells in Response to UVB Irradiation

Photochemistry and Photobiology. May-Jun, 2009  |  Pubmed ID: 19076309

MicroRNAs (miRNAs) are known as a kind of small, noncoding RNA, which play an important role in mediating many biological processes such as development, cell proliferation and differentiation in plants and animals. Here we report the differential expression profiles of miRNAs and characterized putative target genes in NIH3T3 cells at a series of different time points after UVB irradiation (compared with no UVB irradiation). The relative expression of mature miRNA genes was determined by miRNA microarray technique and the results were confirmed by real-time reverse transcriptase polymerase chain reaction (qRT-PCR). Potential target genes of these miRNAs were classified into different function categories with the GOstat software (http://gostat.wehi.edu.au/cgi-bin/goStat.pl). Several miRNAs in this study expressed highly at different time points, especially mmu-miR-365 and mmu-miR-21. Three miRNAs were lowly expressed, of which mmu-miR-465 showed low levels of expression at all time points, whereas after 50 J m(-2) UVB irradiation mmu-miR-296 and mmu-miR-376c showed low levels of expression at 6 and 12 h, respectively. Our study provided a basis for the global characterization of UV-regulated miRNA expression.

[Purification and Characterization of an Antimicrobial Peptide from Paris Polyphylla Var. Chinensis]

Wei Sheng Wu Xue Bao = Acta Microbiologica Sinica. Apr, 2009  |  Pubmed ID: 19621638

We isolated an endophyte PCE45 from the rhizome of Paris polyphylla var. chinensis. From PCE45, we purified and characterized an antimicrobial peptide.

[Construcion of a Chimeric Japanese Encephalits Virus/dengue Virus-2]

Bing Du Xue Bao = Chinese Journal of Virology / [bian Ji, Bing Du Xue Bao Bian Ji Wei Yuan Hui]. May, 2009  |  Pubmed ID: 19634760

The prM/E gene of DV2 was cloned into the JEV (SA14-14-2 strain) replicon vector which had been constructed previously, and the resulting recombinant plasmid was named pPartialdeltaprM/E. The constructed chimeric clone was linearized and then was transcripted into RNA in vitro. The produced RNA was transfected into BHK-21 cells. Five to seven days later, CPE could be observed on the transfected BHK-21cells, and then the supernatant containing the chimeric virus was collected. The Supernatant was inoculated to BHK-1 cells and C6/36 cells, respectively. CPE could be observed about 4 days post the infection of C6/36cell with the chimeric virus. The results from RT-PCR, IFA, Western blot showed that the virus contained the chimeric RNA and the envelop protein of DV2. However, the chimeric virus could not be passaged in BHK-21 cell. The successful construction of the infectious clone JE/DEN-2 laid the basis for the further research of the DV vaccine.

Subcellular Localization of APMCF1 and Its Biological Significance of Expression Pattern in Normal and Malignant Human Tissues

Journal of Experimental & Clinical Cancer Research : CR. 2009  |  Pubmed ID: 19664239

APMCF1 is a novel human gene first cloned from apoptotic MCF-7 cells. Our previous study found ectogenic APMCF1 could induce G1 arrest in hepatocarcinoma cell line HHCC. In order to search its broad expression profile for further understanding of its mechanism in tumor, we investigated a subcellular location of APMCF1 and performed an immunohistochemistry study including various tumor and normal tissues. Discovery from the expression characterization of AMPCF1 may have applicability in the analysis of its biological function in tumor.

[Ultrasound Guided Transrectal Prostate Biopsy: 11-year Experience]

Zhonghua Yi Xue Za Zhi. Apr, 2009  |  Pubmed ID: 19671305

To summarize the experience in ultrasound guided transrectal prostate biopsy and analyze the influencing factors of the biopsy results.

A Ca(2+)-activated Cl(-) Conductance in Interstitial Cells of Cajal Linked to Slow Wave Currents and Pacemaker Activity

The Journal of Physiology. Oct, 2009  |  Pubmed ID: 19703958

Interstitial cells of Cajal (ICC) are unique cells that generate electrical pacemaker activity in gastrointestinal (GI) muscles. Many previous studies have attempted to characterize the conductances responsible for pacemaker current and slow waves in the GI tract, but the precise mechanism of electrical rhythmicity is still debated. We used a new transgenic mouse with a bright green fluorescent protein (copGFP) constitutively expressed in ICC to facilitate study of these cells in mixed cell dispersions. We found that ICC express a specialized 'slow wave' current. Reversal of tail current analysis showed this current was due to a Cl(-) selective conductance. ICC express ANO1, a Ca(2+)-activated Cl(-) channel. Slow wave currents are not voltage dependent, but a secondary voltage-dependent process underlies activation of these currents. Removal of extracellular Ca(2+), replacement of Ca(2+) with Ba(2+), or extracellular Ni(2+) (30 microm) blocked the slow wave current. Single Ca(2+)-activated Cl() channels with a unitary conductance of 7.8 pS were resolved in excised patches of ICC. These are similar in conductance to ANO1 channels (8 pS) expressed in HEK293 cells. Slow wave current was blocked in a concentration-dependent manner by niflumic acid (IC(50) = 4.8 microm). Slow wave currents are associated with transient depolarizations of ICC in current clamp, and these events were blocked by niflumic acid. These findings demonstrate a role for a Ca(2+)-activated Cl(-) conductance in slow wave current in ICC and are consistent with the idea that ANO1 participates in pacemaker activity.

Inhibition of Proliferation, Invasion, and Migration of Prostate Cancer Cells by Downregulating Elongation Factor-1alpha Expression

Molecular Medicine (Cambridge, Mass.). Nov-Dec, 2009  |  Pubmed ID: 19707524

Overexpression of elongation factor-1alpha (EF-1alpha) has been reported to contribute to the development and progression of various cancers. However, its role in prostate cancer (PCa) still remains poorly understood. In the present study, we investigate the influence of EF-1alpha in Du145, a high-grade metastatic PCa cell line, and demonstrate that EF-1alpha plays an essential role in cellular properties associated with tumor progression, namely cell proliferation, invasion, and migration. In this study, EF-1alpha expression in human PCa cell line Du145 was reduced by RNA interference (RNAi) technology, and the proliferation, invasion, and migration of EF-1alpha-reduced Du145 cells were examined. We also detected an EF-1alpha expression pattern in 20 pairs of primary PCa samples and their corresponding normal tissues. Expression of EF-1alpha was detectable in four PCa cell lines (22RV1, LnCap, Du145, and PC3), indicating its possible role in pathogenesis of PCa. RNAi-mediated knockdown of EF-1alpha expression in Du145 cells, which expressed the highest level of EF-1alpha among four PCa cell lines, led to a decrease in proliferation. Similarly, suppression of EF-1alpha inhibited Du145 cell migration and invasion through a basement membrane substitute. Furthermore, we found that the normal prostate tissues showed a relatively low level of EF-1alpha expression, whereas PCa tissues demonstrated significantly higher expression levels of EF-1alpha (P < 0.001). Taken together, these findings support the hypothesis that EF-1alpha affects multiple processes involved in tumor progression, and identify EF-1alpha as a potential therapeutic target.

Endophthalmitis Caused by Vibrio Alginolyticus

Journal of Clinical Microbiology. Oct, 2009  |  Pubmed ID: 19710275

Vibrio alginolyticus is a facultative anaerobic gram-negative bacillus found in normal marine flora. Ocular infections induced by V. alginolyticus are extremely rare. We report a case of endophthalmitis caused by V. alginolyticus to draw attention to V. alginolyticus infections following ocular injuries.

[Influence of Cyclosporine A on Atrial L-type Calcium Channel Alpha1c Subunit in a Canine Model of Atrial Fibrillation]

Zhonghua Xin Xue Guan Bing Za Zhi. Feb, 2009  |  Pubmed ID: 19719984

To investigate the impact of cyclosporine A (CsA) on atrial expression change of L-type calcium channel alpha1c subunit in a canine model of atrial fibrillation (AF).

Experiments on Moving Interaction Boundaries and Their Characteristics of Focusing and Probing of Both Guest and Host Target Molecules

Analytica Chimica Acta. Sep, 2009  |  Pubmed ID: 19720181

In this paper, the concept of a moving interaction boundary (MIB) is proposed with regard to guest and host molecules. With 2-naphthalene-sulfonate (2-NS) and beta-cyclodextrin (CD) as the model guest and host compounds, respectively, the relevant experiments were carried out on the MIB in capillary electrophoresis (CE). The experiments show that (1) there are a MIB and a complex boundary (CB) if proper guest and host molecules are used; (2) the MIB system has the characteristic of selective focusing and probing of the target 2-NS; (3) the system also has the characteristic of selective probing of the target host molecule beta-CD without UV-absorbance, making the direct UV determination of beta-CD from other CDs possible; (4) interestingly, the focusing of the guest molecule is a kind of leaky-sample stacking rather than a collection of analytes in sample sweeping; (5) the mechanism of MIB-induced separation of target analyte from unwanted ones is similar to but different from that of an affinity chromatography. In addition, the utility of MIB was briefly tested for a real sample of wastewater spiked with 2-NS.

Notch1 Induces Enhanced Expression of Delta-like-1 in the U251MG Glioma Cell Line

International Journal of Molecular Medicine. Oct, 2009  |  Pubmed ID: 19724883

The Notch signaling pathway takes part in coordinated regulation of cell growth, survival and differentiation. Previous findings have shown that Notch1 and Delta-like-1 (DLL1) are overexpressed in many glioma cell lines and primary human gliomas. Down-regulation of DLL1 by RNA interference inhibits proliferation and induces apoptosis in multiple glioma cell lines. Our studies showed that Notch1 expression plasmid induced more expression of DLL1 in the U251MG glioma cell line. Adversely, blocking Notch1 receptors down-regulated the expression of DLL1. Both down-regulating DLL1 and blocking Notch1 receptors induced U251MG cell apoptosis and proliferation inhibition, and combining the two treatments produced stronger effects than the sum of a single treatment. These findings suggest a positive feedback loop between Notch1 and DLL1, which may become an effective combined therapeutic target.

GASZ is Essential for Male Meiosis and Suppression of Retrotransposon Expression in the Male Germline

PLoS Genetics. Sep, 2009  |  Pubmed ID: 19730684

Nuage are amorphous ultrastructural granules in the cytoplasm of male germ cells as divergent as Drosophila, Xenopus, and Homo sapiens. Most nuage are cytoplasmic ribonucleoprotein structures implicated in diverse RNA metabolism including the regulation of PIWI-interacting RNA (piRNA) synthesis by the PIWI family (i.e., MILI, MIWI2, and MIWI). MILI is prominent in embryonic and early post-natal germ cells in nuage also called germinal granules that are often associated with mitochondria and called intermitochondrial cement. We find that GASZ (Germ cell protein with Ankyrin repeats, Sterile alpha motif, and leucine Zipper) co-localizes with MILI in intermitochondrial cement. Knockout of Gasz in mice results in a dramatic downregulation of MILI, and phenocopies the zygotene-pachytene spermatocyte block and male sterility defect observed in MILI null mice. In Gasz null testes, we observe increased hypomethylation and expression of retrotransposons similar to MILI null testes. We also find global shifts in the small RNAome, including down-regulation of repeat-associated, known, and novel piRNAs. These studies provide the first evidence for an essential structural role for GASZ in male fertility and epigenetic and post-transcriptional silencing of retrotransposons by stabilizing MILI in nuage.

[Effect of Osteopontin Silencing by Lentivirus-mediated Delivery of SiRNA on Glioma Cell Invasion and Apoptosis]

Zhonghua Yi Xue Yi Chuan Xue Za Zhi = Zhonghua Yixue Yichuanxue Zazhi = Chinese Journal of Medical Genetics. Oct, 2009  |  Pubmed ID: 19806573

To investigate the effect of osteopontin silencing on the invasion and apoptosis of U251 cells.

Culturing of Ventricle Cells at High Density and Construction of Engineered Cardiac Cell Sheets Without Scaffold

International Heart Journal. Sep, 2009  |  Pubmed ID: 19809213

In natural heart tissue, cell density is about 1.0 x 108/cm3, and the cell metabolism is very active. Therefore, culturing heart cells in 3-dimensions at high density and construction of engineered cardiac tissue in vitro is very difficult. The aim of this study was to simulate 3-dimensional culturing of cardiac cells and pursue a novel method to construct engineered cardiac tissue in vitro. The isolated neonatal rat ventricle cells were cultured at a high seeding density of 1 x 10(6)/cm2. The cells at high density metabolized actively; the glucose consumption and lactic acid production of ventricle cells were much greater than those of fibroblasts cultured at the same density. The pH value of the culture medium of ventricle cells consistently decreased more rapidly. These cultured ventricle cells contained vascular endothelial cells, cardiomyocytes, and smooth muscle cells that appeared close to each other, and had overlapping nuclei and plenty of extracellular collagen. The cells at high density were treated with 0.2% trypsin to construct engineered cardiac cell sheets without scaffold. The engineered cardiac cell sheets could beat and roll up spontaneously, each sheet was 3 to 5 cells thick, and contained abundant cardiomyocytes and extracellular collagen. In conclusion, cells cultured at high-density in vitro grew well in a 2-dimensional culturing environment, formed "quasi 3-dimension" culturing, and engineered cardiac cell sheets comprised of several layers of cells were constructed. This study provides some guidance for cardiac tissue engineering and a novel method to construct engineered cardiac tissue without scaffold.

Sex Chromosome Inactivation in the Male

Epigenetics : Official Journal of the DNA Methylation Society. Oct, 2009  |  Pubmed ID: 19838052

Mammalian females have two X chromosomes, while males have only one X plus a Y chromosome. In order to balance X-linked gene dosage between the sexes, one X chromosome undergoes inactivation during development of female embryos. This process has been termed X-chromosome inactivation (XCI). Inactivation of the single X chromosome also occurs in the male, but is transient and is confined to the late stages of first meiotic prophase during spermatogenesis. This phenomenon has been termed meiotic sex chromosome inactivation (MSCI). A substantial portion ( approximately 15-25%) of X-linked mRNA-encoding genes escapes XCI in female somatic cells. While no mRNA genes are known to escape MSCI in males, approximately 80% of X-linked miRNA genes have been shown to escape this process. Recent results have led to the proposal that the RNA interference mechanism may be involved in regulating XCI in female cells. We suggest that some MSCI-escaping miRNAs may play a similar role in regulating MSCI in male germ cells.

Translational Regulation by the 3' Untranslated Region of the Dengue Type 2 Virus Genome

The American Journal of Tropical Medicine and Hygiene. Nov, 2009  |  Pubmed ID: 19861617

The role of the 3'untranslated region (UTR) of the dengue virus (DENV) genome during viral translation remains to be elucidated. We assessed the contribution of well-defined RNA elements in the 3'UTR of DENV-2 to viral translation using a virus-induced reporting gene system and deoxyribozymes (DRzs) targeting the 3'UTR of the DENV-2 genome. Results show that mRNAs carrying a deletion of repeated conserved sequence (RCS2)-CS2 are translated less efficiently than wild type mRNAs. However, mRNAs with a deletion of CS1-stem loop (SL) are translated more efficiently. Thus, CS1-SL and RCS2-CS2 may have different effects on translational regulation. Additionally, the translation-suppressing effect of CS1-SL or the SL element is further confirmed in DENV-2-infected cells using DRzs. Mutagenesis studies show that, rather than the secondary structure, nucleotides 10663-10677 and 10709-10723 are responsible for translational suppression of SL. Overall, our results demonstrate that sequences and elements within the DENV-2 3'UTR regulate viral translation.

[Fluorescent Microbeads-based Multiplex Detection of IgM Antibodies to Pathogens Caused Viral Hemorrhagic Fever]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Apr, 2009  |  Pubmed ID: 20104764

To develop and evaluate a multiplex detection of IgM antibodies to pathogens caused viral hemorrhagic fever.

[Improvement on Primary Culture of Human Nasal Epithelium by Enzymatical Dissociation]

Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi = Journal of Clinical Otorhinolaryngology, Head, and Neck Surgery. Dec, 2009  |  Pubmed ID: 20359105

To highlight the key points of primary culture of human nasal epithelial cells by enzymatical dissociation for high achievement ratio, and to establish a successful primary culture model for subsequent experiments.

Presence and Regulation of Cannabinoid Receptors in Human Retinal Pigment Epithelial Cells

Molecular Vision. 2009  |  Pubmed ID: 19547718

Cannabinoid receptors have been detected in neuron cells and proposed as potential therapeutic agents in neurodegenerative disorders because of their involvement in controlling neural cell survival and death. However, their presence and role in human retinal pigment epithelial (RPE) cells, which play a key role in initiating and developing age related macular degeneration (ARMD), have never been investigated. Here we analyzed the expression of and changes in cannabinoid receptors (CB1 and CB2) and one enzyme responsible for endocannabinoid hydrolysis, fatty acid amide hydrolase (FAAH), in RPE cell oxidative damage process, a cellular model of ARMD.

Pleiotropic Roles of S100A12 in Coronary Atherosclerotic Plaque Formation and Rupture

Journal of Immunology (Baltimore, Md. : 1950). Jul, 2009  |  Pubmed ID: 19542470

Macrophages, cytokines, and matrix metalloproteinases (MMP) play important roles in atherogenesis. The Ca(2+)-binding protein S100A12 regulates monocyte migration and may contribute to atherosclerosis by inducing proinflammatory cytokines in macrophages. We found significantly higher S100A12 levels in sera from patients with coronary artery disease than controls and levels correlated positively with C-reactive protein. S100A12 was released into the coronary circulation from ruptured plaque in acute coronary syndrome, and after mechanical disruption by percutaneous coronary intervention in stable coronary artery disease. In contrast to earlier studies, S100A12 did not stimulate proinflammatory cytokine production by human monocytes or macrophages. Similarly, no induction of MMP genes was found in macrophages stimulated with S100A12. Because S100A12 binds Zn(2+), we studied some functional aspects that could modulate atherogenesis. S100A12 formed a hexamer in the presence of Zn(2+); a novel Ab was generated that specifically recognized this complex. By chelating Zn(2+), S100A12 significantly inhibited MMP-2, MMP-9, and MMP-3, and the Zn(2+)-induced S100A12 complex colocalized with these in foam cells in human atheroma. S100A12 may represent a new marker of this disease and may protect advanced atherosclerotic lesions from rupture by inhibiting excessive MMP-2 and MMP-9 activities by sequestering Zn(2+).

IL-17 Induces Myocardial Fibrosis and Enhances RANKL/OPG and MMP/TIMP Signaling in Isoproterenol-induced Heart Failure

Experimental and Molecular Pathology. Dec, 2009  |  Pubmed ID: 19527710

This study was designed to investigate whether IL-17 can regulate the expression of the MMP/TIMP system, the OPG/RANK/RANKL system, or type-I and type-III collagen fibers in a rat model of isoproterenol-induced heart failure (HF). We also investigated the effect of IL-17 on myocardial fibrosis in this model.

D-Ribosylated Tau Forms Globular Aggregates with High Cytotoxicity

Cellular and Molecular Life Sciences : CMLS. Aug, 2009  |  Pubmed ID: 19517062

Although the glycation of Tau that is involved in paired helical filament formation in Alzheimer's disease has been widely studied, little attention has been paid to the role of D-ribose in the glycation of Tau. Here, we show that Tau is rapidly glycated in the presence of D-ribose, resulting in oligomerization and polymerization. Glycated derivatives appeared after 24 h incubation. Western blotting indicated the formation of advanced glycation end-products (AGEs) during initial stages of glycation. Thioflavin T-positive (ThT-positive) aggregations that appeared from day 4 indicated the globular-like features. Atomic force microscopy revealed that the surface morphology of ribosylated Tau40 was globular-like. Kinetic studies suggested that D-ribosylated Tau is slowly oligomerized and rapidly polymerized with ThT-positive features. Moreover, D-ribosylated Tau aggregates were highly toxic to SHSY5Y cells and resulted in both apoptosis and necrosis. This work has demonstrated that D-ribose reacted with Tau protein rapidly, producing ThT-positive aggregations which had high cytotoxicity.

Male Infertility Caused by Spermiogenic Defects: Lessons from Gene Knockouts

Molecular and Cellular Endocrinology. Jul, 2009  |  Pubmed ID: 19481682

Spermiogenesis refers to the process by which postmeiotic spermatids differentiate into elongated spermatids and eventually spermatozoa. During spermiogenesis, round spermatids undergo dynamic morphologic changes, which include nuclear condensation and elongation, formation of flagella and acrosome, reorganization of organelles and elimination of cytoplasm upon spermiation. This cellular differentiation process is unique to male haploid germ cells, which may explain why approximately half of the testis-specific genes are exclusively expressed in spermiogenesis. The spermiogenesis-specific expression implies that these genes contribute to either structural or functional aspects of future sperm. Many such genes have been inactivated in mice and some of these gene knockout mice display male infertility due to nonfunctional sperm which display no or various degrees of structural abnormalities. Since the majority of these spermiogenesis-specific genes are highly conserved between mice and humans, findings from knockout mouse studies may be applicable to human infertility. Here, I briefly review some of these spermatid-specific gene knockouts. The mouse studies strongly suggest that sperm quality rather than quantity is a better indicator of male fertility and novel assays should be developed to determine sperm functionality.

Determination of Estrogens and Bisphenol A in Bovine Milk by Automated On-line C30 Solid-phase Extraction Coupled with High-performance Liquid Chromatography-mass Spectrometry

Journal of Chromatography. A. Oct, 2009  |  Pubmed ID: 19477451

Using triacontyl bonded silica (C30) as on-line solid-phase extraction (SPE) material and a specially designed on-line analytical system which allowed large sample volume injection, a high speed and robust on-line SPE-HPLC-MS method was established for the analysis of five estrogens and bisphenol A (BPA) in milk samples. The milk sample is pretreated with acetonitrile for protein precipitation and then treated with primary secondary amine for the removal of polar impurities in the matrix. Then the pretreated sample can be automatically loaded by a LC pump. For effective extraction, an offshoot with NH4Ac solution of high-flow rate was employed to dilute the loaded sample by a mixing tee before sample was loaded onto the C30 extraction column. In this way, large volume injection (1 mL in this experiment) could be achieved. Some important parameters such as sample loading flow rate, sample dilution ratio and injection volume were optimized. Under the optimized conditions, the recoveries for all analytes range from 71.4 to 97.1% and reproducibility represented as RSDs are less than 15.0% (n=5) with milk samples spiked at 0.6 and 15 ng/mL of each analyte. To the authors' knowledge, it constitutes the first work describing a C30 on-line SPE-LC-MS analytical method for the screen and monitoring of these estrogens and BPA in milk.

Fixure-reduce Method for the Synthesis of Cu2O/MWCNTs Nanocomposites and Its Application As Enzyme-free Glucose Sensor

Biosensors & Bioelectronics. Jul, 2009  |  Pubmed ID: 19473828

Cu(2)O/MWCNT (multi-walled carbon nanotubes) nanocomposites were successfully prepared in large quantities with a new fixure-reduction method under low temperature. The morphology and shape of the Cu(2)O/MWCNTs nanocomposites were characterized by field emission scanning electron microscopes (FESEMs), energy dispersive X-ray (EDX), X-ray photoelectron spectroscopy (XPS) and X-ray powder diffraction (XRD), respectively. Cyclic voltammetry (CV) was used to evaluate the electrochemical performance of the Cu(2)O/MWCNTs nanocomposites modified electrode towards glucose. Compared to the bare GCE, the Cu(2)O nanoparticles and the MWCNTs modified electrode, the Cu(2)O/MWCNTs modified electrode displays high electrocatalytic activity towards the oxidation of glucose. With a potential of -0.20 V, the Cu(2)O/MWCNTs modified electrode was used to determine glucose by amperometric, showing significantly lower overvoltage and a linear dependence (R=0.9958) in the concentration up to 10 microM with a sensitivity of 6.53 microA micromol L(-1) and a detection limit of 0.05 micromol L(-1) (signal-to-noise ratio of 3). In summary, the preparation process of the nanocomposites is very simple and the nanocomposites could be used for the development of enzyme-free glucose sensor.

The Expression of NF-E2-related Factor 2 in the Rat Brain After Traumatic Brain Injury

The Journal of Trauma. May, 2009  |  Pubmed ID: 19430250

Secondary brain damage plays a critical role in the outcome of patients with traumatic brain injury (TBI). The mechanisms underlying secondary brain damage are complex. A target that can interrupt multiple mechanisms underlying secondary brain damage may represent a promising new therapeutic approach for TBI. NF-E2-related factor 2 (Nrf2) is the key regulator in reducing oxidative stress, inflammatory damage, and the accumulation of toxic metabolites, which are all involved in secondary brain damage after TBI. Therefore, Nrf2 might represent a new direction for the treatment of TBI. However, the expression pattern of Nrf2 after TBI has not yet been studied.

The Protective Effect of the Ketogenic Diet on Traumatic Brain Injury-induced Cell Death in Juvenile Rats

Brain Injury : [BI]. May, 2009  |  Pubmed ID: 19408168

The ketogenic diet (the KD) is an effective treatment for intractable epilepsy, especially in the paediatric population, and a growing number of studies have shown the neuroprotective role of the KD. However, few studies focused on the neuroprotective effects of the KD in traumatic brain injury (TBI). The present study aimed to investigate the effects of the KD on TBI.

[Role of Focal Adhesion Kinase in the Hypoxia-induced Invasion of SMMC-7721 Cells]

Zhonghua Gan Zang Bing Za Zhi = Zhonghua Ganzangbing Zazhi = Chinese Journal of Hepatology. Apr, 2009  |  Pubmed ID: 19403028

To study focal adhesion kinase (FAK) expression in hypoxia-stressed SMMC-7721 cells and the role of FAK expression in the hypoxia-induced invasion of SMMC-7721 cells.

[Secreted Expression of Dengue Virus Type I Envelope Glycoprotein in 293T Cells]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Dec, 2009  |  Pubmed ID: 20718342

To expression prM/E gene of dengue virus type I in mammalia cells.

[Identification and Construction of Replicon Vectors of Japanese Encephalitis Virus (strain SA14-14-2)]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Dec, 2009  |  Pubmed ID: 20718343

In order to lay the groundwork for studying the novel vaccine Identified.

Combined Sulphur Cycle Based System of Hydrogen Production and Biological Treatment of Wastewater

Environmental Technology. Nov, 2009  |  Pubmed ID: 19950472

The experiment was conducted to investigate continuous hydrogen production with lower cost and sulphate-rich wastewater treatment. In this paper, both anaerobic bio-treatment of sulphate-rich wastewater and hydrogen production were applied to construct a laboratory-scale combined sulphur cycle based system. The system consisted of two reactors, which were a photocatalytic reactor and an anaerobic bioreactor, respectively. In the anaerobic bioreactor, sulphate-reducing bacteria (SRB) converted SO4(2-) to S(2-). The produced S(2-) yielded by SRB was further used as a sacrificial reagent to produce H2 in the photocatalytic reactor. Then, S(2-) was changed into SO4(2-), which returned to the anaerobic bioreactor for treatment again. The present study highlighted an advantage compared with the conventional method, in that no extra S(2-) was added to the photocatalytic reactor, which reduced the total cost and realized continuous hydrogen production. The average COD removal efficiency was 79.6%.

2-(2-Pyridylsulfan-yl)acetic Acid

Acta Crystallographica. Section E, Structure Reports Online. 2009  |  Pubmed ID: 21580116

All non-H atoms of the title compound, C(7)H(7)NO(2)S, lie on a crystallographic mirror plane, with the two methyl-ene H atoms bis-ected by this plane. The crystal packing is characterized by inter-molecular C-H⋯O and O-H⋯N contacts, which link the mol-ecules into infinite zigzag chains parallel to [010].

Enzymatic Degradation Products from a Marine Polysaccharide YCP with Different Immunological Activity and Binding Affinity to Macrophages, Hydrolyzed by Alpha-amylases from Different Origins

Biochimie. Apr, 2010  |  Pubmed ID: 20004229

YCP is a marine polysaccharide with anti-tumor and immune-modulating effects. This study evaluated the effect of enzymatic degradation of YCP by alpha-amylases from different origins on its immunological activity and binding ability to the macrophages. YCP was hydrolyzed by alpha-amylases isolated from Aspergillus oryzae, Bacillus licheniformis, Barley malt, and Porcine pancreas respectively, then four fragments with unique molecular weight (termed: YCP-Ao, YCP-Bl, YCP-Bm, and YCP-Pp, respectively) were obtained. The four fragments showed different immunological activity and the ability to bind to macrophages. Among them, YCP-Ao possessed almost equivalent immunological activity compared to the original YCP, while such properties were not retained in YCP-Bl. Our further study showed that YCP-Ao prevented YCP from binding to macrophages. In conclusion, YCP-Ao and YCP might have similar active regions.

NF-κB, a Hot Topic in Biochemical and Medical Studies in China

Science China. Life Sciences. Dec, 2010  |  Pubmed ID: 21181352

[Relationships of Blood Stasis Syndrome, CYP2C19 Gene Polymorphism with Clopidogrel Resistance and Post-PCI Prognosis]

Zhongguo Zhong Xi Yi Jie He Za Zhi Zhongguo Zhongxiyi Jiehe Zazhi = Chinese Journal of Integrated Traditional and Western Medicine / Zhongguo Zhong Xi Yi Jie He Xue Hui, Zhongguo Zhong Yi Yan Jiu Yuan Zhu Ban. Dec, 2010  |  Pubmed ID: 21302482

To study the relationships of blood stasis syndrome (BSS), CYP2C19 gene polymorphism with clopidogrel resistance (CR) and post-PCI prognosis.

[Effect of Lutein on Relieving Oxidative Stress in Mice Induced by D-galatose]

Wei Sheng Yan Jiu = Journal of Hygiene Research. Jul, 2010  |  Pubmed ID: 20726230

To study the effect of lutein on relieving oxidative stress in the liver of mice induced by D-galactose(D-gal).

Zinc Finger-zinc Finger Interaction Between the Transcription Factors, GATA-1 and Sp1

Biochemical and Biophysical Research Communications. Oct, 2010  |  Pubmed ID: 20807505

In contrast to the extensive understanding of the zinc finger-DNA interactions, less is known about zinc finger-zinc finger interactions. GATA-1 and Sp1 are transcription factors with zinc finger domains for DNA binding. The interaction between the GATA-1 and Sp1 zinc finger domains is important for synergistic transcriptional effects in erythroid genes. Despite the biological importance of the GATA-1 and Sp1 interaction, the molecular mechanism of the interaction remains unclear. We constructed a series of deletion mutants of the zinc finger domains of GATA-1 and Sp1 to identify the regions within the GATA-1 and Sp1 zinc finger domains that interact. The zinc finger-zinc finger interaction modes were also estimated from calorimetric measurements. This revealed that the interaction between the Sp1 and GATA-1 zinc finger domains was primarily electrostatic, and that the linker region of the Sp1 zinc fingers is important for the association with the GATA-1 zinc finger domains. We propose a new molecular mechanism for zinc finger-zinc finger interactions that should contribute to our understanding of the bio-functional role of the interaction between GATA-1 and Sp1.

A Decade of Clinical Experience with Extra-adrenal Paragangliomas of Retroperitoneum: Report of 67 Cases and a Literature Review

Urology Annals. Jan, 2010  |  Pubmed ID: 20842251

The purpose was to highlight the diagnosis and treatment of extra-adrenal para-gangliomas, which often causes catecholamine hypersecretion and hypertension.

[Analysis of Misdiagnosis of Calcifying Epithelioma in Oral and Maxillofacial-head and Neck Region]

Hua Xi Kou Qiang Yi Xue Za Zhi = Huaxi Kouqiang Yixue Zazhi = West China Journal of Stomatology. Aug, 2010  |  Pubmed ID: 20848947

Eight patients misdiagnosed as calcifying epithelioma during 2003-2008 at Department of Oral and Maxillofacial Surgery of Cangzhou Central Hospital were retrospectively analyzed. The pertinent literatures were reviewed and the pathogenesis, clinical manifestations, imaging performance and differential diagnosis of calcifying epithelioma were discussed.

Elevated Levels of Tau Protein in Cerebrospinal Fluid of Patients with Probable Creutzfeldt-Jakob Disease

The American Journal of the Medical Sciences. Oct, 2010  |  Pubmed ID: 20881758

A definitive diagnosis of Creutzfeldt-Jakob disease (CJD) can only be made by neuropathologic examination and demonstration of typical pathologic changes and the pathologic prion protein in central nervous tissues. This study investigated the diagnostic sensitivity and specificity of the microtubule-association protein tau in cerebrospinal fluid (CSF) from Chinese patients with sporadic CJD.

Inactivation of PI3K/AKT Signaling Inhibits Glioma Cell Growth Through Modulation of β-catenin-mediated Transcription

Brain Research. Dec, 2010  |  Pubmed ID: 20888802

Aberrant Wnt/β-catenin signaling contributes to the development of many cancers, including glial tumorigenesis. While cross talk between the Wnt/β-catenin and PI3K/AKT signaling pathways has been proposed, the impact of PI3K/AKT inhibition on β-catenin signaling in glioma remains unknown. In the present study, we report decreased cell proliferation and invasive ability upon the LY294002-induced inhibition of PI3K in both U251 and LN229 human glioblastoma cells in vitro. Pharmacologic inhibition of PI3K resulted in the downregulation of several members of the β-catenin pathway, including Fra-1, c-Myc, and cyclin D1. Downregulation impacted β-catenin-mediated transcription, as LY294002 decreased β-catenin/TCF transcriptional activity, determined by the reporter assay. Similar results were observed in vivo, as intratumoral injection of LY294002 downregulated the expression of the components of the β-catenin pathway and delayed tumor growth in nude mice harboring subcutaneous LN229 xenografts. These results suggest that the PI3K/AKT signaling pathway regulates glioma cell proliferation, in part via repression of the Wnt/β-catenin pathway.

[Effect of Bone Morphogenetic Protein 4 on Glioma Stem Cell Proliferation and Apoptosis in Vitro]

Zhonghua Yi Xue Yi Chuan Xue Za Zhi = Zhonghua Yixue Yichuanxue Zazhi = Chinese Journal of Medical Genetics. Oct, 2010  |  Pubmed ID: 20931521

To investigate the role of bone morphogenetic protein 4 (BMP4) on the proliferation and apoptosis in glioma stem cells.

The Role of Nrf2 Signaling in the Regulation of Antioxidants and Detoxifying Enzymes After Traumatic Brain Injury in Rats and Mice

Acta Pharmacologica Sinica. Nov, 2010  |  Pubmed ID: 20953205

To determine whether Nrf2 signaling pathway activation could attenuate oxidative stress and neuronal damage following traumatic brain injury (TBI).

Porcine Circovirus Type 2 (PCV2): Genetic Variation and Newly Emerging Genotypes in China

Virology Journal. 2010  |  Pubmed ID: 20955622

Porcine circovirus type 2 (PCV2), the causative agent of postweaning multisystemic wasting syndrome (PMWS), is a serious economic problem for the swine industry in China. In this study, we investigated the genetic variation of PCV2 in China using strains isolated from 2004-2008. Viruses were isolated from samples collected from pigs with multi-systemic lesions and clinical signs of PMWS from different regions of China, and the genomes of these viruses were sequenced. The assembled sequences were used to define the genotypes of these strains; PCR-RFLP methodology was used to distinguish isolates and capture ELISA was used to demonstrate the antigenic changes resulted from ORF2 gene mutation of the isolates.

[A Pilot Study of Weekly Versus 3-week Docetaxel in Combination with Capecitabine in Patients with Anthracycline-pretreated Metastatic Breast Cancer]

Zhonghua Yi Xue Za Zhi. Jul, 2010  |  Pubmed ID: 20979824

To evaluate the efficacy and safety of weekly or 3-week docetaxel in combination with capecitabine.

Very Low Density Lipoprotein Receptor Subtype II Silencing by RNA Interference Inhibits Cell Proliferation in Hepatoma Cell Lines

Hepato-gastroenterology. Jul-Aug, 2010  |  Pubmed ID: 21033246

Very low density lipoprotein receptor (VLDLR) belongs to the low density lipoprotein receptor family, it is divided into two subtypes according to forms with an absence (type II) or a presence (type I) of the O-linked sugar domain. VLDLR have been detected in kinds of cancers so far; however, the subtype of VLDLR in hepatocellular carcinoma (HCC) tissues and hepatoma cell lines has yet to be reported. We detected the VLDLR expression in 39 cases of hepatocellular carcinoma and in three kinds of hepatoma cell lines: HepG2, HBV transfected HepG2.2.15, SMMC-7721 and normal human fetal liver cell line LO2 using RT-PCR and western blotting. The results showed that both type I and type II VLDLR were detected in HCC tissues and hepatoma cell lines, and the type II VLDLR expression was significantly higher than that of type I in cell lines. We inhibited the type II VLDLR expression by shRNA-mediated RNA interference in HepG2, SMMC-7721 cell and then subsequently found the cell proliferation slowed down. The cyclinD1 expression confirmed the cell cycle was arrested at the G0/G1 phase, suggesting that inhibiting the type II VLDLR expression may have a positive impact on carcinogenesis of HCC.

[Correction of Secondary Deformity of Unilateral Cleft Lip with Refined Anatomic Repair in Orbicularis Oris Muscle]

Zhonghua Zheng Xing Wai Ke Za Zhi = Zhonghua Zhengxing Waike Zazhi = Chinese Journal of Plastic Surgery. Jul, 2010  |  Pubmed ID: 21046769

To investigate the functional repair of secondary deformity of unilateral cleft lip.

[A Two-year Follow-up Study on the Efficacy of Ursodeoxycholic Acid on Primary Biliary Cirrhosis in Different Stages]

Zhonghua Gan Zang Bing Za Zhi = Zhonghua Ganzangbing Zazhi = Chinese Journal of Hepatology. Oct, 2010  |  Pubmed ID: 21059288

To assess the therapeutic effect of primary biliary cirrhosis(PBC) in different stages with ursodeoxycholic acid (UDCA).

Liver X Receptor Agonist Methyl-3β-hydroxy-5α,6α-epoxycholanate Attenuates Atherosclerosis in Apolipoprotein E Knockout Mice Without Increasing Plasma Triglyceride

Pharmacology. 2010  |  Pubmed ID: 21071998

Liver X receptors (LXRs) promote macrophage reverse cholesterol transport and cholesterol excretion from the body. The synthetic LXR ligands T0901317 and GW3965 were shown to significantly inhibit atherosclerosis in mice and to increase the expression of ATP-binding cassette transporter A1 (ABCA1) in the atherosclerotic lesions. However, these compounds increase plasma and hepatic triglyceride (TG) levels in mice. Methyl-3β-hydroxy-5α,6α-epoxycholanate (MHEC), synthesized from hyodeoxycholic acid, functions as an LXR agonist, but its role in atherogenesis and lipid metabolism remained to be elucidated.

[Disorder of Tiangui (kidney Essence) and Reproductive Dysfunction in Patients with Polycystic Ovary Syndrome]

Zhong Xi Yi Jie He Xue Bao = Journal of Chinese Integrative Medicine. Nov, 2010  |  Pubmed ID: 21078264

Traditional Chinese medicine (TCM) usually views polycystic ovary syndrome (PCOS) as a menstrual disease or infertility disease. Reproductive dysfunction in PCOS is characterized by ovarian androgen excess and disturbance of follicular development, and its main clinical manifestations include delayed menstruation, scant menstruation, amenorrhea or infertility. Insulin resistance is a key pathological mechanism of PCOS. "Tiangui" (kidney essence) as a sex-stimulating essence in female in TCM theory, is essential to the menstruation and pregnancy of women. The disturbance of Tiangui (including time, status and rhythm) would result in female reproductive problems. Current studies of Tiangui indicate that ovary is the target organ of PCOS treatment, and its functional characteristics are consistent with the properties of Tiangui in time frame, state form and rhythm cycle. It is then concluded that ovarian dysfunction in PCOS can be expressed as disorder of Tiangui.

FOXP3 Expression and Clinical Characteristics of Hepatocellular Carcinoma

World Journal of Gastroenterology : WJG. Nov, 2010  |  Pubmed ID: 21086571

To study the biological and clinical characteristics of transcription factor forkhead box protein 3 (FOXP3) in hepatocellular carcinoma (HCC).

In Vitro and in Vivo Changes to PLGA/sirolimus Coating on Drug Eluting Stents

Biomaterials. Jul, 2010  |  Pubmed ID: 20382420

The degradation behavior of the absorbable coating may have significant impacts to the short and long-term safety of the drug eluting stent (DES). A poly(lactide-co-glycolide) (PLGA) polymer coating containing sirolimus on stent surface was studied when stents were expanded to 3.0 mm. In vitro and in vivo degradation behavior of coated stents and cast films were characterized by light microscope, gel permeation chromatography (GPC), weight loss, field-emission environmental scanning electron microscope (ESEM-FEG) and SEM combined with energy dispersive spectrometer (EDS). For in vivo study, specially designed chambers with semi-permeable membrane were used to host the stents during the implantation. The data indicated the coating polymer lost 80% molecular weight and 60% of its mass in 60 days. The complete degradation of the coating occurred in 6 months. The degradation of the coating seemed to be homogeneous, and the coating went through a hydration/swelling process before it became completely degraded. It was noted that the coating maintained its integrity and strong adhesion to the strut during the entire degradation process, no fragmentation delaminate were observed. The study also demonstrated similar degradation behaviors of the drug coating in in vitro and in vivo conditions.

Small RNA Cloning

Methods in Molecular Biology (Clifton, N.J.). 2010  |  Pubmed ID: 20387155

The next generation sequencing technologies have made the exhaustive sequencing of all small RNA species within a small RNA library possible. Thus, comparative studies on the changes in small-RNA transcriptomes between two biological samples represent a powerful means of revealing the functions of small RNAs in physiological and pathological processes. Here, we describe a small RNA cloning method that can be used to generate small RNA cDNA libraries for both conventional and next generation (deep) sequencing. It can also be used for detection and quantitative analyses of small RNAs using PCR.

In Situ Hybridization Detection of MicroRNAs

Methods in Molecular Biology (Clifton, N.J.). 2010  |  Pubmed ID: 20387156

MicroRNAs (miRNAs) are endogenous approximately 22 nucleotide RNAs that play critical roles in many cellular processes including cell differentiation, proliferation, and apoptosis. The analysis of spatiotemporal expression of miRNAs is important for dissecting their roles in development and during physiological/pathophysiological processes. In situ hybridization is a powerful technology that allows cellular localization. However, the detection of miRNAs by in situ hybridization has been challenging because of the low affinity of conventional RNA or DNA probes due to the small sizes of miRNAs. Here, we describe a protocol for miRNA in situ hybridization on mouse tissue cryosections using locked nucleic acid (LNA) probes. LNA probes demonstrate a much higher affinity to their complimentary RNAs compared to conventional RNA and DNA oligo probes, which allow detection of miRNAs in tissue sections with excellent specificity.

Detection and Quantitative Analysis of Small RNAs by PCR

Methods in Molecular Biology (Clifton, N.J.). 2010  |  Pubmed ID: 20387157

Increasing lines of evidence indicate that small non-coding RNAs including miRNAs, piRNAs, rasiRNAs, 21U endo-siRNAs, and snoRNAs are involved in many critical biological processes. Functional studies of these small RNAs require a simple, sensitive, and reliable method for detecting and quantifying levels of small RNAs. Here, we describe such a method that has been widely used for the validation of cloned small RNAs and also for quantitative analyses of small RNAs in both tissues and cells.

[The Effects of Lycopene on Reactive Oxygen Species and Anoxic Damage in Ischemia Reperfusion Injury in Rats]

Zhonghua Yu Fang Yi Xue Za Zhi [Chinese Journal of Preventive Medicine]. Jan, 2010  |  Pubmed ID: 20388361

To study the protective effects of lycopene (LP) on cerebral ischemia-reperfusion injury induced by focal cerebral ischemia and oxidative stress in rats.

Enhancing Antibody Fc Heterodimer Formation Through Electrostatic Steering Effects: Applications to Bispecific Molecules and Monovalent IgG

The Journal of Biological Chemistry. Jun, 2010  |  Pubmed ID: 20400508

Naturally occurring IgG antibodies are bivalent and monospecific. Bispecific antibodies having binding specificities for two different antigens can be produced using recombinant technologies and are projected to have broad clinical applications. However, co-expression of multiple light and heavy chains often leads to contaminants and pose purification challenges. In this work, we have modified the CH3 domain interface of the antibody Fc region with selected mutations so that the engineered Fc proteins preferentially form heterodimers. These novel mutations create altered charge polarity across the Fc dimer interface such that coexpression of electrostatically matched Fc chains support favorable attractive interactions thereby promoting desired Fc heterodimer formation, whereas unfavorable repulsive charge interactions suppress unwanted Fc homodimer formation. This new Fc heterodimer format was used to produce bispecific single chain antibody fusions and monovalent IgGs with minimal homodimer contaminants. The strategy proposed here demonstrates the feasibility of robust production of novel Fc-based heterodimeric molecules and hence broadens the scope of bispecific molecules for therapeutic applications.

Extended Dual-grating Alignment Method for Optical Projection Lithography

Applied Optics. Feb, 2010  |  Pubmed ID: 20119023

Since accurate alignment is essential for projection lithography, an extended dual-grating based alignment scheme is proposed. This method is an extension of the basic dual-grating alignment model, the mechanism of which is explained to make it clear how the extended scheme performs in projection lithography. The framework of the extended alignment scheme for projection lithography is constructed, and the process of key parameter determination is then detailed. In both cases, a tiny shift of the wafer during the alignment process can be resolved by a conspicuous displacement or phase variation of corresponding fringes. Analytical results indicate that alignment is independent of the gap between wafer and mask, disturbance from the fluctuation in illumination can be neglected, and alignment resolution in subnanometers can be realized with this scheme.

Ribosylation Rapidly Induces Alpha-synuclein to Form Highly Cytotoxic Molten Globules of Advanced Glycation End Products

PloS One. 2010  |  Pubmed ID: 20140223

Alpha synuclein (alpha-Syn) is the main component of Lewy bodies which are associated with several neurodegenerative diseases such as Parkinson's disease. While the glycation with D-glucose that results in alpha-Syn misfold and aggregation has been studied, the effects of glycation with D-ribose on alpha-Syn have not been investigated.

Effect of MgO Contents on the Mechanical Properties and Biological Performances of Bioceramics in the MgO-CaO-SiO2 System

Journal of Materials Science. Materials in Medicine. May, 2010  |  Pubmed ID: 20162324

The aim of this research was to investigate the effect of the chemical composition on the mechanical properties, bioactivity, and cytocompatibility in vitro of bioceramics in the MgO-CaO-SiO(2) system. Three single-phase ceramics (merwinite, akermanite and monticellite ceramics) with different MgO contents were fabricated. The mechanical properties were tested by an electronic universal machine, while the bioactivity in vitro of the ceramics was detected by investigating the bone-like apatite-formation ability in simulated body fluid (SBF), and the cytocompatibility was evaluated through osteoblast proliferation and adhesion assay. The results showed that their mechanical properties were improved from merwinite to akermanite and monticellite ceramics with the increase of MgO contents, whereas the apatite-formation ability in SBF and cell proliferation decreased. Furthermore, osteoblasts could adhere, spread and proliferate on these ceramic wafers. Finally, the elongated appearance and minor filopodia of cells on merwinite ceramic were more obvious than the other two ceramics.

GATA3 Inhibits Breast Cancer Metastasis Through the Reversal of Epithelial-mesenchymal Transition

The Journal of Biological Chemistry. Apr, 2010  |  Pubmed ID: 20189993

GATA3, a transcription factor that regulates T lymphocyte differentiation and maturation, is exclusively expressed in early stage well differentiated breast cancers but not in advanced invasive cancers. However, little is understood regarding its activity and the mechanisms underlying this differential expression in cancers. Here, we employed GATA3-positive, non-invasive (MCF-7) and GATA3-negative, invasive (MDA-MB-231) breast cancer cells to define its role in the transformation between these two distinct phenotypes. Ectopic expression of GATA3 in MDA-MB-231 cells led to a cuboidal-like epithelial phenotype and reduced cell invasive activity. These cells also increased E-cadherin expression but decreased levels of vimentin, N-cadherin, and MMP-9. Further, MDA-MB-231 cells expressing GATA3 grew smaller primary tumors without metastasis compared with larger metastatic tumors derived from control MDA-MB-231 cells in xenografted mice. GATA3 was found to induce E-cadherin expression through binding GATA-like motifs located in the E-cadherin promoter. Blockade of GATA3 using small interfering RNA gene knockdown in MCF-7 cells triggered fibroblastic transformation and cell invasion, resulting in distant metastasis. Studies of human breast cancer showed that GATA3 expression correlated with elevated E-cadherin levels, ER expression, and long disease-free survival. These data suggest that GATA3 drives invasive breast cancer cells to undergo the reversal of epithelial-mesenchymal transition, leading to the suppression of cancer metastasis.

Electrochemical Immunosensor Based on Colloidal Carbon Sphere Array

Biosensors & Bioelectronics. Jan, 2010  |  Pubmed ID: 19879126

A novel type of colloidal carbon sphere array (CSA) was developed for the fabrication of disposable electrochemical immunosensor. The CSA was successfully prepared on indium tin oxide (ITO) substrate in a simple manner and the scanning electron micrograph confirmed that a single-layered arrangement of the carbon spheres with its (111) plane paralleled the substrate's surface. The CSA modified electrode has a higher surface area and exhibits a more sensitive electrochemical response than a normal carbon-based electrode with the same geometric area. An Immunoglobin A (IgA) immunosensor was constructed by the covalent bonding of IgA antibody molecules with the CSA aided by large numbers of carboxyl groups on the surface of carbon spheres. The immunosensor exhibited a wide linear response to IgA ranging from 0.1 to 200 ng mL(-1) by electrochemical impedance spectroscopy (EIS) technique. The detection of IgA levels in three sera obtained from hospital samples showed acceptable accuracy.

A Model to Study the Phenotypic Changes of Interstitial Cells of Cajal in Gastrointestinal Diseases

Gastroenterology. Mar, 2010  |  Pubmed ID: 19917283

Interstitial cells of Cajal (ICC) express the receptor tyrosine kinase, KIT, the receptor for stem cell factor. In the gastrointestinal (GI) tract, ICC are pacemaker cells that generate spontaneous electrical slow waves, and mediate inputs from motor neurons. Absence or loss of ICC are associated with GI motility disorders, including those consequent of diabetes. Studies of ICC have been hampered by the low density of these cells and difficulties in recognizing these cells in cell dispersions.

[Effects of Lycopene on Cerebral Ischemia-reperfusion Injury in Rats]

Wei Sheng Yan Jiu = Journal of Hygiene Research. Mar, 2010  |  Pubmed ID: 20459036

To study the protective effects of lycopene (LP) on cerebral ischemia-reperfusion injury and oxidative stress in SD rats and the mechanism of them.

Expression Pattern of Osteopontin Splice Variants and Its Functions on Cell Apoptosis and Invasion in Glioma Cells

Neuro-oncology. Aug, 2010  |  Pubmed ID: 20511184

Osteopontin (OPN) is widely overexpressed in various cancers, including gliomas, and plays an important role in tumorigenesis. However, the expression pattern and functions of OPN splice variants expressed in gliomas remain unclear. The aims of our current study were to examine the expression pattern and functions of OPN splice variants in gliomas. In present study, the mRNA levels of OPN splice variants are markedly increased in gliomas tissues, and all OPN splice variants were also found in U251 and U87 cells. Furthermore, knock-down and regain of function experiments were designed to explore the functions of OPN splice variants in U251 and U87 cells. Lentiviral vectors of OPN small interference RNA (siRNA) targeting all three endogenous mRNAs of OPN and OPN splice variants synonymous mutant that were not silenced by OPN siRNA were constructed. Our results showed that all OPN splice variants synonymous mutant-protected glioma cells from apoptosis induced by OPN siRNA through alteration of the levels of Bcl-2 family proteins and OPN-b Mu elicted a significant effect. Both OPN-a Mu and -c Mu promoted glioma cell invasion through alteration of the levels of uPA, MMP-2, and MMP-9 expressions and the activities of MMP-2 and MMP-9 via activation PI-3K/AKT/NF-kappaB signaling pathway. Moreover, OPN-c Mu showed the strongest effect on glioma cell invasion, while OPN-b Mu showed no effect on the invasion of U251 and U87 cells. Thus, different splice variants of OPN have divergent functions in regulating apoptosis and invasion of glioma cells, which broadens their importance in glioma biotherapy.

T-wave Oversensing and Inappropriate Shocks in Implantable Cardioverter Defibrillators

Chinese Medical Journal. May, 2010  |  Pubmed ID: 20529568

An Experimental Study of the Population and Evolutionary Dynamics of Vibrio Cholerae O1 and the Bacteriophage JSF4

Proceedings. Biological Sciences / The Royal Society. Nov, 2010  |  Pubmed ID: 20538647

Studies of Vibrio cholerae in the environment and infected patients suggest that the waning of cholera outbreaks is associated with rise in the density of lytic bacteriophage. In accordance with mathematical models, there are seemingly realistic conditions where phage predation could be responsible for declines in the incidence of cholera. Here, we present the results of experiments with the El Tor strain of V. cholerae (N16961) and a naturally occurring lytic phage (JSF4), exploring the validity of the main premise of this model: that phage predation limits the density of V. cholerae populations. At one level, the results of our experiments are inconsistent with this hypothesis. JSF4-resistant V. cholerae evolve within a short time following their confrontation with these viruses and their populations become limited by resources rather than phage predation. At a larger scale, however, the results of our experiments are not inconsistent with the hypothesis that bacteriophage modulate outbreaks of cholera. We postulate that the resistant bacteria that evolved play an insignificant role in the ecology or pathogenicity of V. cholerae. Relative to the phage-sensitive cells from whence they are derived, the evolved JSF4-resistant V. cholerae have fitness costs and other characters that are likely to impair their ability to compete with the sensitive cells in their natural habitat and may be avirulent in human hosts. The results of this in vitro study make predictions that can be tested in natural populations of V. cholerae and cholera-infected patients.

Effect of Temperature on the Population Growth of Rhynchophorus Ferrugineus (Coleoptera: Curculionidae) on Sugarcane

Environmental Entomology. Jun, 2010  |  Pubmed ID: 20550815

The effect of temperature on the developmental time, survival, and reproduction of Rhynchophorus ferrugineus (Olivier) reared on sugarcane was studied at seven constant temperatures (16, 20, 24, 28, 32, 36, and 40 degrees C). The developmental threshold temperatures and effective accumulated temperatures for the whole generation were 17.41 degrees C and 1,590.72 DD, respectively. One generation had the highest survival rate (26.67%) at 28 degrees C. The egg failed to survive at 16 and 40 degrees C. The population trend index (I = 38.22) and net reproductive rate (R(o) = 38.3) were highest at 28 degrees C. The net reproductive rate (R(o) = 3.36), intrinsic rate of increase (r(m) = 0.0028), and finite capacity of increase (lambda = 1.0028) were lowest at 20 degrees C. The mean generation time (T(o) = 85.82) was shortest at 36 degrees C. The population double time (PDT = 27.08) was shortest at 32 degrees C. Based on these studies, we concluded that the temperatures from 28 to 32 degrees C were the most suitable temperatures for the development of R. ferrugineus.

UBQLN1 Interacts with SPEM1 and Participates in Spermiogenesis

Molecular and Cellular Endocrinology. Oct, 2010  |  Pubmed ID: 20558241

Spermiogenesis represents the process through which haploid male germ cells differentiate from round spermatids into elongated spermatids and eventually the male gametes called spermatozoa. Haploid cell differentiation is unique to male germ cell development and many unique genes/proteins essential for this process have been discovered. SPEM1 is one of these spermiogenesis-essential proteins encoded by a testis-specific gene exclusively expressed in the developing spermatids. Inactivation of Spem1 in mice results in deformed spermatozoa characterized by "head-bent-back" abnormalities with 100% penetrance. Using yeast two-hybrid screening assays, we identified UBQLN1 as one of the SPEM1-interacting partners. UBQLN1 and SPEM1 were colocalized to the manchette of elongating spermatids. Since UBQLN1 functions through binding and directing poly-ubiquitinated proteins to the proteasome for degradation, interactions between UBQLN1 and SPEM1 suggest a role in the regulation of protein ubiquitination during spermiogenesis.

Post-draw PAN-PMMA Nanofiber Reinforced and Toughened Bis-GMA Dental Restorative Composite

Dental Materials : Official Publication of the Academy of Dental Materials. Sep, 2010  |  Pubmed ID: 20579722

The polyacrylonitrile (PAN)-poly(methyl methacrylate) (PMMA) core-shell nanofiber reinforced dental composites have been investigated for their excellent interface adhesive, and this kind of novel dental composite has the potential for clinical uses such as denture base resin and crown-bridge material. The first objective of this work was to determine the improving effect of tensile properties by post-drawing PAN-PMMA nanofibers membrane. The second objective was to examine the flexural strength (Fs), flexural modulus (Ey) and work of fracture (WOF) of Bis-GMA/TEGDMA composites reinforced with PAN-PMMA nanofibers.

[Effect of Phosphatase PHLPP1 Gene Transfer on the Proliferation of Human Umbilical Vein Endothelial Cells]

Nan Fang Yi Ke Da Xue Xue Bao = Journal of Southern Medical University. Jun, 2010  |  Pubmed ID: 20584661

To investigate the constituent expression of PH domain leucine-rich repeat protein phosphatase 1 (PHLPP1) in human umbilical vein endothelial cells (HUVECs) and the effect of PHLPP1 gene transfer on the proliferation of the cells in vitro.

Zmynd15 Encodes a Histone Deacetylase-dependent Transcriptional Repressor Essential for Spermiogenesis and Male Fertility

The Journal of Biological Chemistry. Oct, 2010  |  Pubmed ID: 20675388

Spermatogenesis is a complex process through which male germ line stem cells undergo a multi-step differentiation program and sequentially become spermatogonia, spermatocytes, spermatids, and eventually spermatozoa. In this process, transcription factors act as switches that precisely regulate the expression of genes that in turn control the developmental program of male germ cells. Transcription factors identified to be essential for normal haploid gene expression all display transcription-activating effects and thus serve as the "on" switch for haploid gene expression. Here, we report that ZMYND15 acts as a histone deacetylase-dependent transcriptional repressor and controls normal temporal expression of haploid cell genes during spermiogenesis. Inactivation of Zmynd15 results in early activation of transcription of numerous important haploid genes including Prm1, Tnp1, Spem1, and Catpser3; depletion of late spermatids; and male infertility. ZMYND15 represents the first transcriptional repressor identified to be essential for sperm production and male fertility.

[Construction of Eukaryotic Vector of Small Hairpin Interfering RNA Against NYD-SP5]

Zhonghua Nan Ke Xue = National Journal of Andrology. May, 2010  |  Pubmed ID: 20684327

NYD-SP5 is a newly cloned gene highly expressed in human testes, which consists of 3 598 nucleotides including a 1 027-amino acid open reading frame. It is a human-mouse homologous gene. The domain analysis indicated that the NYD-SP5 protein is a transmembrane protein. This study aimed to design and establish recombinant plasmids of small hairpin interfering RNA (shRNA) against NYD-SP5, and to pave the way for the analysis of the function of NYD-SP5 in the testis using the transgenic mouse model.

Room-temperature Ionic Liquid Assisted Fabrication of Sensitive Electrochemical Immunosensor Based on Ordered Macroporous Gold Film

The Analyst. Oct, 2010  |  Pubmed ID: 20694205

A novel label-free highly sensitive electrochemical impedance spectroscopy (EIS) immunosensor was fabricated based on the highly ordered macroporous gold film (HOMGF) in the presence of room-temperature ionic liquid (IL) for the detection of human Apolipoprotein B-100 (ApoB-100). The antibody of ApoB-100 (Ab) was adsorbed directly onto the HOMGF electrode surface and maintained its bioactivity. After the residual active sites at the electrode were passivated by BSA, the mixture of BMIm(+)BF(4)(-) and silica sol was dropped onto the electrode to entrap the adsorbed Ab and BSA molecules firmly. The addition of IL could prevent the inactivation of Ab by releasing alcohol during the sol-gel process, and the conductivity of the IL-gel membrane was increased. Of particular interest is the fact that the fabricated immunosensor could be used at 60 °C. This could be attributed to the interconnected porosity of the IL-gel membrane, which can prevent Ab from unfolding and losing its bioactivities. The immunosensor also exhibited a highly sensitive response to ApoB-100 with the lowest concentration of 5 fg mL(-1). The detection of ApoB-100 levels in five sera samples obtained from hospital showed acceptable accuracy with that using commercial immunonephelometry method.

Malondialdehyde Suppresses Cerebral Function by Breaking Homeostasis Between Excitation and Inhibition in Turtle Trachemys Scripta

PloS One. 2010  |  Pubmed ID: 21203547

The levels of malondialdehyde (MDA) are high in the brain during carbonyl stress, such as following daily activities and sleep deprivation. To examine our hypothesis that MDA is one of the major substances in the brain leading to fatigue, the influences of MDA on brain functions and neuronal encodings in red-eared turtle (Trachemys scripta) were studied. The intrathecal injections of MDA brought about sleep-like EEG and fatigue-like behaviors in a dose-dependent manner. These changes were found associated with the deterioration of encoding action potentials in cortical neurons. In addition, MDA increased the ratio of γ-aminobutyric acid to glutamate in turtle's brain, as well as the sensitivity of GABAergic neurons to inputs compared to excitatory neurons. Therefore, MDA, as a metabolic product in the brain, may weaken cerebral function during carbonyl stress through breaking the homeostasis between excitatory and inhibitory neurons.

Human Skeletal Muscle-derived Stem Cells Retain Stem Cell Properties After Expansion in Myosphere Culture

Experimental Cell Research. Apr, 2011  |  Pubmed ID: 21277299

Human skeletal muscle contains an accessible adult stem-cell compartment in which differentiated myofibers are maintained and replaced by a self-renewing stem cell pool. Previously, studies using mouse models have established a critical role for resident stem cells in skeletal muscle, but little is known about this paradigm in human muscle. Here, we report the reproducible isolation of a population of cells from human skeletal muscle that is able to proliferate for extended periods of time as floating clusters of rounded cells, termed "myospheres" or myosphere-derived progenitor cells (MDPCs). The phenotypic characteristics and functional properties of these cells were determined using reverse transcription-polymerase chain reaction (RT-PCR), flow cytometry and immunocytochemistry. Our results showed that these cells are clonogenic, express skeletal progenitor cell markers Pax7, ALDH1, Myod, and Desmin and the stem cell markers Nanog, Sox2, and Oct3/4 significantly elevated over controls. They could be maintained proliferatively active in vitro for more than 20 weeks and passaged at least 18 times, despite an average donor-age of 63 years. Individual clones (4.2%) derived from single cells were successfully expanded showing clonogenic potential and sustained proliferation of a subpopulation in the myospheres. Myosphere-derived cells were capable of spontaneous differentiation into myotubes in differentiation media and into other mesodermal cell lineages in induction media. We demonstrate here that direct culture and expansion of stem cells from human skeletal muscle is straightforward and reproducible with the appropriate technique. These cells may provide a viable resource of adult stem cells for future therapies of disease affecting skeletal muscle or mesenchymal lineage derived cell types.

Coxiella Burnetii Antigen-stimulated Dendritic Cells Mediated Protection Against Coxiella Burnetii in BALB/c Mice

The Journal of Infectious Diseases. Jan, 2011  |  Pubmed ID: 21288829

Coxiella burnetii is the etiological agent of human Q fever. In this study, adaptive transfer of mouse bone marrow-derived dendritic cells (BMDCs) stimulated with C. burnetii antigen, phase I whole-cell antigen (PIAg), lipopolysaccharide (LPS)-removed PIAg (PIIAg), protein antigen Com1, or SecB significantly reduced coxiella burden in recipient mice compared with control mice. Mice that received PIIAg-pulsed BMDCs displayed substantially lower coxiella burden than recipient mice of PIAg-pulsed BMDCs after C burnetii challenge. The protection offered by the antigen-activated BMDCs was correlated with the increased proliferation of helper T (T(H)) T(H)1 CD4(+) cells, preferential development of T(H)17 cells, and impaired expansion of regulatory T lymphocytes. Our results suggest that PIIAg is far superior to PIAg in activating BMDCs to confer protection against C. burnetii in vivo, whereas Com1 and SecB are protective antigens because Com1- or SecB-pulsed BMDCs confer partial protection.

[The Operation Time and Post-operative Target Eye Alignment of Intermittent Exotropia]

[Zhonghua Yan Ke Za Zhi] Chinese Journal of Ophthalmology. Nov, 2011  |  Pubmed ID: 22336059

Different from other types of strabismus, children with intermittent exotropia can maintain normal eye alignment and binocular vision in a long period of time. But the course of intermittent exotropia is mostly progressive, the eye position may drift outward even after surgery, so there is a certain risk of recurrence or undercorrection. These features often lead confusion to the clinicians in determining the operation time and postoperative target eye position. So how to choose the appropriate intervention time and target eye aligment is a challenge for clinicians. In this paper, we will combine the research status and clinical practice, discuss these issues, and present some point of view for the peer reference.

[Surgical Treatment of Myopic Strabismus Fixus by Modified Yokoyama's Surgery]

[Zhonghua Yan Ke Za Zhi] Chinese Journal of Ophthalmology. Nov, 2011  |  Pubmed ID: 22336061

To evaluate the surgical results of modified Yokoyama's procedure for treating myopic strabismus fixus.

[The Progress of Intermittent Exotropia]

[Zhonghua Yan Ke Za Zhi] Chinese Journal of Ophthalmology. Nov, 2011  |  Pubmed ID: 22336071

Intermittent exotropia is a type of strabismus between exophoria and constant exotropia. The onset age is usually one to four years old. Different from other types of strabismus, intermittent exotropia patient can maintain normal binocular vision in a long period of time. So how to choose the optimal timing and effective treatment is a challenge for clinicians. The objective of this article is to review the recent research progress of intermittent exotropia, including the severity classification, non-surgical treatment, timing of surgery and surgical methods, overcorrection and undercorrection treatment etc., and give some recommendations for the treatment of intermittent exotropia.

The Characteristics of the Synonymous Codon Usage in Hepatitis B Virus and the Effects of Host on the Virus in Codon Usage Pattern

Virology Journal. Dec, 2011  |  Pubmed ID: 22171933

ABSTRACT: BACKGROUND: Hepatitis B virus (HBV) infection is one of the main human health problem and causes a large-scale of patients chronic infection worldwide.. As the replication of HBV depends on its host cell system, codon usage pattern for the viral gene might be susceptible to two main selections, namely mutation pressure and translation selection. In this case, a deeper investigation between HBV evolution and host adaptive response might assist control this disease. Result: Relative synonymous codon usage (RSCU) values for the whole HBV coding sequence were studied by Principal component analysis (PCA). The characteristics of the synonymous codon usage patterns, nucleotide contents and the comparison between ENC values of the whole HBV coding sequence indicated that the interaction between virus mutation pressure and host translation selection exists in the processes of HBV evolution. The synonymous codon usage pattern of HBV is a mixture of coincidence and antagonism to that of host cell. But the difference of genetic characteristic of HBV failed to be observed to its different epidemic areas or subtypes, suggesting that geographic factor is limited to influence the evolution of this virus, while genetic characteristic based on HBV genotypes could be divided into three groups, namely (i) genotyps A and E, (ii) genotype B, (iii) genotypes C, D and G. CONCLUSION: Codon usage patterns from PCA for identification of evolutionary trends in HBV provide an alternative approach to understand the evolution of HBV. Further more, a combined selection of mutation pressure with translation selection on codon usage might shed a light on understanding the evolutionary trends of HBV genotypes.

[Investigations on Cyclotorsion Changes Following Strabismus Surgery in Superior Oblique Palsy Patients]

[Zhonghua Yan Ke Za Zhi] Chinese Journal of Ophthalmology. Sep, 2011  |  Pubmed ID: 22177124

To investigate cyclotorsion changes after strabismus surgery in superior oblique palsy patients.

[Effects of Infection and Stress on Transient Receptor Potential Vanilloid Receptor 1 and Extracellular-regulated Kinase in Spinal Cord of Mast Cell Deficient Rats]

Beijing Da Xue Xue Bao. Yi Xue Ban = Journal of Peking University. Health Sciences. Dec, 2011  |  Pubmed ID: 22178825

To investigate the effects of mast cells (MCs) and the relationship between the signal pathway including transient receptor potential vanilloid receptor 1 (TRPV1) and extracellular-regulated kinase (ERK) and MCs in rat models of visceral hyperalgesia triggered by infection and stress.

STK31(TDRD8) is Dynamically Regulated Throughout Mouse Spermatogenesis and Interacts with MIWI Protein

Histochemistry and Cell Biology. Dec, 2011  |  Pubmed ID: 22205278

Tudor-domain-containing proteins (TDRDs) are suggested to be critical regulators of germinal granules assembly involved in Piwi-interacting RNAs (piRNAs)-mediated pathways, of which associated components and the underlying functional mechanisms, however, remain to be elucidated. We herein characterized the expression pattern of STK31, a member of TDRDs subfamily (also termed as TDRD8), throughout spermatogenesis during mouse postnatal development. RT-PCR and Western blot verified its preferential expression in testis, but not in any other somatic tissues, in addition to embryonic stem cells. Immunofluorescent staining demonstrated that STK31 was confined to granules-like structures in mid-to-late spermatocyte cytoplasm and to acrosomal cap starting at steps 7-8 of spermatids. Furthermore, STK31 retained its localization to equatorial segment of acrosome during epididymal maturation, capacitation, and acrosome reaction. Co-immunoprecipitation assay in vivo and in vitro confirmed MIWI is a bona fide partner of STK31 in mice testes, in combination with LC/MS identification. We also discovered a group of heat shock proteins specifically associated with STK31 in vivo. Our findings suggest mouse STK31 could be a potential nuage-associated protein in the cytoplasm of mid-to-late spermatocytes and play pivotal roles related to fertilization.

Argininosuccinate Synthase Conditions the Response to Acute and Chronic Ethanol-induced Liver Injury in Mice

Hepatology (Baltimore, Md.). Dec, 2011  |  Pubmed ID: 22213272

BACKGROUND AND AIM: Argininosuccinate synthase (ASS) is the rate-limiting enzyme in both the urea and the L-citrulline/nitric oxide (NO·) cycles regulating protein catabolism, ammonia levels and NO· generation (1-2). Since a proteomics analysis identified ASS and nitric oxide synthase-2 (NOS2) as co-induced in rat hepatocytes by chronic ethanol consumption, which also occurred in alcoholic liver disease (ALD) and in cirrhotic patients, we hypothesized that ASS could play a role in ethanol binge and chronic ethanol-induced liver damage. METHODS: To investigate the contribution of ASS to the pathophysiology of ALD, wild-type (WT) and Ass(+/-) mice (Ass(-/-) are lethal due to hyperammonemia) were exposed to an ethanol binge or to chronic ethanol drinking. RESULTS: Compared with WT, Ass(+/-) mice given an ethanol binge exhibited decreased steatosis, lower NOS2 induction and less 3-nitrotyrosine (3-NT) protein residues, indicating that reducing nitrosative stress via the L-citrulline/NO· pathway plays a significant role in preventing liver damage. However, chronic ethanol treated Ass(+/-) mice displayed enhanced liver injury compared with WT mice. This was due to hyperammonemia, lower phosphorylated AMP-activated protein kinase (pAMPKα) to total AMPKα ratio, decreased sirtuin (Sirt-1) and peroxisomal proliferator-activated receptor coactivator-1α (Pgc1α) mRNAs, lower fatty acid β-oxidation due to down-regulation of carnitine palmitoyl transferase-II (CPT-II), decreased antioxidant defense and elevated lipid peroxidation end-products in spite of comparable nitrosative stress but likely reduced NOS3. CONCLUSION: Partial Ass ablation protects only in acute ethanol-induced liver injury by decreasing nitrosative stress but not in a more chronic scenario where oxidative stress and impaired fatty acid β-oxidation are key events. (HEPATOLOGY 2011.).

Expression Pattern of Enkephalinergic Neurons in the Developing Spinal Cord Revealed by Preproenkephalin-green Fluorescent Protein Transgenic Mouse and Its Colocalization with GABA Immunoreactivity

Cells, Tissues, Organs. 2011  |  Pubmed ID: 21124002

To gain better insight into the ontogenic function of enkephalin (ENK) in the spinal cord, it is necessary to have a clear picture of the developing pattern of the ENKergic neurons. To address this question, we used transgenic mice which reveal ENKergic neurons easily by expressing enhanced green fluorescent protein (GFP) under the specific transcriptional control of the preproenkephalin (PPE) gene. GFP-positive neurons first appeared at embryonic day (E) 11.5 in the ventromedial part of the cervical ventral gray matter. At E13, they were mainly present in the intermediate zone. Thereafter, GFP-positive neurons increased progressively in number and extended from ventral to dorsal regions. Quantitative analysis showed that GFP-positive neurons peaked in number at postnatal day (P) 7 at the cervical level. The number of GFP-positive neurons reached a peak at P3 at the lumbar level. At P21, the distribution pattern of GFP immunoreactivity was similar to that in the adult spinal cord. Double-labeling results showed that about one-third of the total γ-aminobutyric acid cell population colocalized with GFP: 34.9 ± 3.5% at E16 and 32.4 ± 3.7% at P3. Double-labeled neurons accounted for nearly half of the GFP-positive neurons: 42.4 ± 2.4% at E16 and 44.1 ± 2.9% at P3. Taken together, the present results suggest that ENKergic neurons develop according to a rostrocaudal and ventrodorsal gradient. These results have broad implications for understanding the functional roles of ENKergic neurotransmission in the developing spinal cord.

Precursor-directed Fungal Generation of Novel Halogenated Chaetoglobosins with More Preferable Immunosuppressive Action

Chemical Communications (Cambridge, England). Feb, 2011  |  Pubmed ID: 21152613

Precursor-fed cultivation of endophytic Chaetomium globosum 1C51 afforded nine novel "unnatural" halogenated chaetoglobosins including those with more preferable immunosuppressive activity.

Index-ion Triggered MS2 Ion Quantification: a Novel Proteomics Approach for Reproducible Detection and Quantification of Targeted Proteins in Complex Mixtures

Molecular & Cellular Proteomics : MCP. Mar, 2011  |  Pubmed ID: 21169564

Biomedical research requires protein detection technology that is not only sensitive and quantitative, but that can reproducibly measure any set of proteins in a biological system in a high throughput manner. Here we report the development and application of a targeted proteomics platform termed index-ion triggered MS2 ion quantification (iMSTIQ) that allows reproducible and accurate peptide quantification in complex mixtures. The key feature of iMSTIQ is an approach called index-ion triggered analysis (ITA) that permits the reproducible acquisition of full MS2 spectra of targeted peptides independent of their ion intensities. Accurate quantification is achieved by comparing the relative intensities of multiple pairs of fragment ions derived from isobaric targeted peptides during MS2 analysis. Importantly, the method takes advantage of the favorable performance characteristics of the LTQ-Orbitrap, which include high mass accuracy, resolution, and throughput. As such it provides an attractive targeted proteomics tool to meet the demands of systems biology research and biomarker studies.

PEGylated Poly(amidoamine) Dendrimer-based Dual-targeting Carrier for Treating Brain Tumors

Biomaterials. Jan, 2011  |  Pubmed ID: 20934215

A dual-targeting drug carrier (PAMAM-PEG-WGA-Tf) based on the PEGylated fourth generation (G = 4.0) PAMAM dendrimer with transferrin (Tf) and wheat germ agglutinin (WGA) on the periphery and doxorubicin (DOX) loaded in the interior was synthesized and its BBB penetration and tumor targeting properties were explored. DLS and TEM measurements revealed the size of PAMAM-PEG-WGA-Tf was in the range of 14-20 nm. It reduced the cytotoxicity of DOX to the normal cells greatly, while efficiently inhibited the growth rate of the C6 glioma cells. The assay of transport across the BBB showed that PAMAM-PEG-WGA-Tf delivered 13.5% of DOX in a period of 2 h, demonstrating an enhanced transport ratio as compared to the ratio of 8% for PAMAM-PEG-WGA, 7% for PAMAM-PEG-Tf and 5% for free DOX in the same period of time. The accumulation of DOX in the tumor site was increased due to the targeting effects of both Tf and WGA, leading to the complete breakage of the avascular C6 glioma spheroids in vitro.

Microarray Analysis of Temperature-induced Transcriptome of Streptococcus Suis Serotype 2

Vector Borne and Zoonotic Diseases (Larchmont, N.Y.). Mar, 2011  |  Pubmed ID: 20795872

Streptococcus suis serotype 2 (S. suis S2) is able to cause human infections ranging from superficial wounded skin infections to severe invasive infections such as meningitis and streptococcal toxic shock-like syndrome. During its infection cycle, S. suis S2 must acclimatize itself to temperature shift. Herein, a whole-genome DNA microarray was used to investigate the global transcriptional regulation of an invasive strain of S. suis S2 grown to late-exponential phase at 29°C or 40°C relative to 37°C. The differentially regulated genes that were detected included those encoding virulence factors, antigenic proteins, ATP-binding-cassette transporters, and proteins of unknown functions. Our data provided a global profile of gene transcription induced by temperature alteration and shed light on some unforeseen lines for further pathogenesis investigation.

Evidence for Epithelial-mesenchymal Transition in Cancer Stem Cells of Head and Neck Squamous Cell Carcinoma

PloS One. 2011  |  Pubmed ID: 21304586

Initiation, growth, recurrence, and metastasis of head and neck squamous cell carcinomas (HNSCC) have been related to the behavior of cancer stem cells (CSC) that can be identified by their aldehyde-dehydrogenase-isoform-1 (ALDH1) activity. We quantified and enriched ALDH1(+) cells within HNSCC cell lines and subsequently characterized their phenotypical and functional properties like invasion capacity and epithelial-mesenchymal transition (EMT). Spheroid culture enriched CSC from five HNSCC cell lines by up to 5-fold. In spheroid-derived cells (SDC) and the parental monolayer-derived cell line ALDH1, CD44, CD24, E-Cadherin, α-SMA, and Vimentin expression was compared by flow-cytometry and immunofluorescence together with proliferation and cell cycle analysis. Invasion activity was evaluated by Matrigel assay and expression of stemness-related transcription factors (TF) Nanog, Oct3/4, Sox2 and EMT-related genes Snail1 and 2, and Twist by real-time PCR. All cell lines formed spheroids that could self-renew and be serially re-passaged. ALDH1 expression was significantly higher in SDC. ALDH1(+) cells showed increased colony-formation. The proportion of cells with a putative CSC marker constellation of CD44(+)/CD24(-) was highly variable (0.5% to 96%) in monolayer and spheroid cultures and overlapped in 0%-33% with the CD44(+)/CD24(-)/ALDH1(+) cell subset. SDC had significantly higher invading activity. mRNA of the stemness-related genes Sox2, Nanog, and Oct3/4 was significantly increased in SDC of all cell lines. Twist was significantly increased in two while Snail2 showed a significant increase in one and a significant decrease in SDC of two cell lines. SDC had a higher G0 phase proportion, showed high-level expression of α-SMA and Vimentin, but significantly decreased E-Cadherin expression. HNSCC-lines harbor potential CSC, characterized by ALDH1 and stemness marker TF expression as well as properties like invasiveness, quiescence, and EMT. CSC can be enriched by anchorage-independent culture techniques, which may be important for the investigation of their contribution to therapy resistance, tumor recurrence and metastasis.

The Population Dynamics of Bacteria in Physically Structured Habitats and the Adaptive Virtue of Random Motility

Proceedings of the National Academy of Sciences of the United States of America. Mar, 2011  |  Pubmed ID: 21325053

Why is motility so common in bacteria? An obvious answer to this ecological and evolutionary question is that in almost all habitats, bacteria need to go someplace and particularly in the direction of food. Although the machinery required for motility and chemotaxis (acquiring and processing the information needed to direct movement toward nutrients) are functionally coupled in contemporary bacteria, they are coded for by different sets of genes. Moreover, information that resources are more abundant elsewhere in a habitat would be of no value to a bacterium unless it already had the means to get there. Thus, motility must have evolved before chemotaxis, and bacteria with flagella and other machinery for propulsion in random directions must have an advantage over bacteria relegated to moving at the whim of external forces alone. However, what are the selection pressures responsible for the evolution and maintenance of undirected motility in bacteria? Here we use a combination of mathematical modeling and experiments with Escherichia coli to generate and test a parsimonious and ecologically general hypothesis for the existence of undirected motility in bacteria: it enables bacteria to move away from each other and thereby obtain greater individual shares of resources in physically structured environments. The results of our experiments not only support this hypothesis, but are quantitatively and qualitatively consistent with the predictions of our model.

Effect of Polydatin on Action Potential in Ventricular Papillary Muscle of Rat and the Underlying Ionic Mechanism

Sheng Li Xue Bao : [Acta Physiologica Sinica]. Feb, 2011  |  Pubmed ID: 21340434

It is proved that polydatin has cardioprotection against ischemia-induced arrhythmia, but the electrophysiological mechanism is not clear. The aim of the present study was to investigate the effect of polydatin on action potential (AP) in ventricular papillary muscle and the underlying ionic mechanism in rat using intracellular recording and whole-cell patch clamp techniques. The results showed: (1) In normal papillary muscles, polydatin (50 and 100 µmol/L) shortened duration of 50% repolarization (APD(50)) and duration of 90% repolarization (APD(90)) in a concentration-dependent manner (P<0.01). But polydatin had no effects on resting potential (RP), overshoot (OS), amplitude of action potential (APA) and maximal rate of depolarization in phase 0 (V(max)) in normal papillary muscles (P>0.05). (2) In partially depolarized papillary muscles, polydatin (50 µmol/L) not only shortened APD(50) and APD(90) (P<0.05), but also decreased OS, APA and V(max) (P<0.05). (3) After pretreatment with glibenclamide (10 µmol/L), an ATP-sensitive K(+) channel blocker, the electrophysiological effect of polydatin (50 µmol/L) was partially inhibited. (4) Pretreatment with N(G)-nitro-L-arginine methyl ester (L-NAME, 1 mmol/L), a nitric oxide (NO) synthase inhibitor, failed to abolish the effect of polydatin (50 µmol/L) on AP. (5) Polydatin (25, 50, 75 and 100 µmol/L) decreased L-type Ca(2+) current in ventricular myocytes in a concentration-dependent manner (P<0.05). (6) Polydatin (50 µmol/L) increased ATP-sensitive K(+) current in ventricular myocytes (P<0.05). The results suggest that polydatin can shorten the repolarization of AP in normal papillary muscle and inhibit AP in partially depolarized papillary muscle, which might be related to the blocking of L-type Ca(2+) channel and the opening of ATP-sensitive K(+) channel.

A YKL-40-neutralizing Antibody Blocks Tumor Angiogenesis and Progression: a Potential Therapeutic Agent in Cancers

Molecular Cancer Therapeutics. May, 2011  |  Pubmed ID: 21357475

Accumulating evidence has indicated that expression levels of YKL-40, a secreted glycoprotein, were elevated in multiple advanced human cancers. Recently, we have identified an angiogenic role of YKL-40 in cancer development. However, blockade of the function of YKL-40, which implicates therapeutic value, has not been explored yet. Our current study sought to establish a monoclonal anti-YKL-40 antibody as a neutralizing antibody for the purpose of blocking tumor angiogenesis and metastasis. A mouse monoclonal anti-YKL-40 antibody (mAY) exhibited specific binding with recombinant YKL-40 and with YKL-40 secreted from osteoblastoma cells MG-63 and brain tumor cells U87. In the functional analysis, we found that mAY inhibited tube formation of microvascular endothelial cells in Matrigel induced by conditioned medium of MG-63 and U87 cells, as well as recombinant YKL-40. mAY also abolished YKL-40-induced activation of the membrane receptor VEGF receptor 2 (Flk-1/KDR) and intracellular signaling mitogen-activated protein (MAP) kinase extracellular signal-regulated kinase (Erk) 1 and Erk 2. In addition, mAY enhanced cell death response of U87 line to γ-irradiation through decreased expression of pAKT and AKT and accordingly, abrogated angiogenesis induced by the conditioned medium of U87 cells in which YKL-40 levels were elevated by treatment with γ-irradiation. Furthermore, treatment of xenografted tumor mice with mAY restrained tumor growth, angiogenesis, and progression. Taken together, this study has shown the therapeutic use for the mAY in treatment of tumor angiogenesis and metastasis.

Role of YKL-40 in the Angiogenesis, Radioresistance, and Progression of Glioblastoma

The Journal of Biological Chemistry. Apr, 2011  |  Pubmed ID: 21385870

Glioblastoma is one of the most fatal cancers, characterized by a strong vascularized phenotype. YKL-40, a secreted glycoprotein, is overexpressed in patients with glioblastomas and has potential as a novel tumor biomarker. The molecular mechanisms of YKL-40 in glioblastoma development, however, are poorly understood. Here, we aimed to elucidate the role YKL-40 plays in the regulation of VEGF expression, tumor angiogenesis, and radioresistance. YKL-40 up-regulated VEGF expression in glioblastoma cell line U87, and both YKL-40 and VEGF synergistically promote endothelial cell angiogenesis. Interestingly, long term inhibition of VEGF up-regulated YKL-40. YKL-40 induced coordination of membrane receptor syndecan-1 and integrin αvβ5, and triggered a signaling cascade through FAK(397) to ERK-1 and ERK-2, leading to elevated VEGF and enhanced angiogenesis. In addition, γ-irradiation of U87 cells increased YKL-40 expression that protects cell death through AKT activation and also enhances endothelial cell angiogenesis. Blockade of YKL-40 activity or expression decreased tumor growth, angiogenesis, and metastasis in xenografted animals. Immunohistochemical analysis of human glioblastomas revealed a correlation between YKL-40, VEGF, and patient survival. These findings have shed light on the mechanisms by which YKL-40 promotes tumor angiogenesis and malignancy, and thus provide a therapeutic target for tumor treatment.

RANBP17 is Localized to the XY Body of Spermatocytes and Interacts with SPEM1 on the Manchette of Elongating Spermatids

Molecular and Cellular Endocrinology. Feb, 2011  |  Pubmed ID: 21184802

We identified Ran-binding protein 17 (RANBP17) as one of the interacting partners of sperm maturation 1 (SPEM1) using yeast 2-hybrid screening and immunoprecipitation assays. Expression profiling analyses suggested that RANBP17 was preferentially expressed in the testis. Immunofluorescent confocal microscopy revealed a dynamic localization pattern of RANBP17 during spermatogenesis. In primary spermatocytes RANBP17 was mainly localized to the XY body. In the subsequent spermiogenesis, RANBP17 was first observed in the nuclei of round spermatids (steps 1-7) and then confined to the manchette of elongating spermatids (steps 8-14) together with its interacting partner SPEM1. In the Spem1-null testes, levels of RANBP17 were significantly elevated. As a member of a large protein family involved in the nucleocytoplasmic transport, RANBP17 may have a role in sex chromosome inactivation during the meiotic phase of spermatogenesis, and also in the intramanchette transport during spermiogenesis. Interactions between RANBP17 and SPEM1, for the first time, point to a potential function of SPEM1 in the RANBP17-mediated nucleocytoplasmic transport.

Amelioration of Cisplatin-induced Nephrotoxicity by Pravastatin in Mice

Experimental and Toxicologic Pathology : Official Journal of the Gesellschaft Für Toxikologische Pathologie. Mar, 2011  |  Pubmed ID: 20060696

This study investigated the protective effects of pravastatin against cisplatin-induced nephrotoxicity and the possible mechanisms in mice. Pravastatin showed significant protection as evidenced by the decrease of elevated serum creatinine (CRE) and blood urea nitrogen (BUN), and improvement of histopathological injury induced by cisplatin. The formation of kidney malondialdehyde (MDA) with a concomitant reduction of reduced glutathione (GSH) were inhibited by pravastatin, while the activities of kidney superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-px) were increased. The over expressions of kidney induced nitric oxide synthase (iNOS) and nitrotyrosine (3-NT) were suppressed by pravastatin. Pravastatin suppressed cisplatin-induced p38 mitogen-activated protein kinase (MAPK) activation in the kidney of mice. These results suggest that pravastatin pre-administration can prevent the nephrotoxicity induced by cisplatin. Pravastatin may exert the protective effect via inhibiting oxidative and nitrosative stress.

Clinical Correlation of MGMT Protein Expression and Promoter Methylation in Chinese Glioblastoma Patients

Medical Oncology (Northwood, London, England). Mar, 2011  |  Pubmed ID: 21394635

Promoter methylation of O6-methylguanine-DNA methyltransferase (MGMT) gene has been considered as a prognostic maker and increasingly emphasized in the treatment of glioblastoma multiforme (GBM). Contrastingly, the correlation of MGMT with clinical outcomes in Chinese glioblastoma patients has not been elucidated systematically. In the present study, tumor tissues from 172 GBM patients were analyzed for MGMT protein expression by immunohistochemistry. Of these, 79 were also subjected to pyrosequencing for MGMT promoter methylation analysis. MGMT protein overexpression was found in 109/172 (63.4%) GBM samples. And no significant survival difference was observed between the patients with MGMT overexpression and low expression in terms of progression-free survival or overall survival (P = 0.605 and P = 0.565, respectively). Meanwhile, MGMT promoter methylation was detected in 26/79 cases (32.9%), whereas 53/79 (67.1%) samples were unmethylated. Further survival analysis also revealed that MGMT promoter methylation status cannot predict patients progression-free survival and overall survival (P = 0.906 and P = 0.548, respectively). The integrated analysis showed that there was significant negative correlation between MGMT protein expression and promoter methylation (P = 0.004). These results underscore that, in Chinese GBM patients, (a) MGMT protein expression level was not a prognostic factor, (b) overall survival but not progression-free survival showed a trend toward increase in patients with MGMT promoter methylation, although the difference was not significant statistically and this observation has to be validated in larger patients cohort, (c) there was a significant correlation between MGMT protein expression in immunohistochemistry and MGMT promoter methylation by pyrosequencing.

Acetic Acid and Lithium Chloride Effects on Hydrothermal Carbonization of Lignocellulosic Biomass

Bioresource Technology. May, 2011  |  Pubmed ID: 21411315

As a renewable non-food resource, lignocellulosic biomass has great potential as an energy source or feedstock for further conversion. However, challenges exist with supply logistics of this geographically scattered and perishable resource. Hydrothermal carbonization treats any kind of biomass in 200 to 260°C compressed water under an inert atmosphere to produce a hydrophobic solid of reduced mass and increased fuel value. A maximum in higher heating value (HHV) was found when 0.4 g of acetic acid was added per g of biomass. If 1g of LiCl and 0.4 g of acetic acid were added per g of biomass to the initial reaction solution, a 30% increase in HHV was found compared to the pretreatment with no additives, along with greater mass reduction. LiCl addition also reduces reaction pressure. Addition of acetic acid and/or LiCl to hydrothermal carbonization each contribute to increased HHV and reduced mass yield of the solid product.

MicroRNA-10b Induces Glioma Cell Invasion by Modulating MMP-14 and UPAR Expression Via HOXD10

Brain Research. May, 2011  |  Pubmed ID: 21419107

MicroRNAs are small endogenous noncoding RNAs, which modulate target gene expression by binding with target mRNA sequences in the 3'untranslated region (UTR) with an imperfect complementarity that inhibits the mRNA translation. Many microRNAs have been reported to function as tumor oncogenes or anti-oncogenes. Recently, more and more microRNAs have been reported to contribute to a tumor's invasive potential. Here, we show that microRNA-10b (miR-10b) was over-expressed in glioma samples and directly associated with the glioma's pathological grade and malignancy. We also found that miR-10b induced glioma cell invasion by modulating tumor invasion factors MMP-14 and uPAR expression via the direct target HOXD10. The miR-10b/HOXD10/MMP-14/uPAR signaling pathway might contribute to the invasion of glioma. Accordingly, glioma cells lost their invasive ability when treated with specific antisense oligonucleotides (miR-10b inhibitors), suggesting that miR-10b could be used as a new bio-target to cure glioma.

Electrochemical Impedance Determination of Polychlorinated Biphenyl Using a Pyrenecyclodextrin-decorated Single-walled Carbon Nanotube Hybrid

Chemical Communications (Cambridge, England). May, 2011  |  Pubmed ID: 21451857

This work reports the first detailed study on an electrochemical impedance sensor for determination of polychlorinated biphenyl (PCB), such as 3,3',4,4'-tetrachlorobiphenyl (PCB-77), based on a single-walled carbon nanotube/pyrenecyclodextrin (SWCNT/PyCD) hybrid.

Serum Response Factor-dependent MicroRNAs Regulate Gastrointestinal Smooth Muscle Cell Phenotypes

Gastroenterology. Jul, 2011  |  Pubmed ID: 21473868

Smooth muscle cells (SMCs) change phenotypes under various pathophysiological conditions. These changes are largely controlled by the serum response factor (SRF), a transcription factor that binds to CC (A/T)6 GG (CArG) boxes in SM contractile genes. MicroRNAs (miRNA) regulate transitions among SMC phenotypes. The SMC miRNA transcriptome (SMC miRNAome) and its regulation by SRF have not been determined.

[Effects of Verapamil on the Proliferation, DNA Synthesis and Migration of Human Hepatoma Cell Line HepG2]

Sichuan Da Xue Xue Bao. Yi Xue Ban = Journal of Sichuan University. Medical Science Edition. Mar, 2011  |  Pubmed ID: 21500559

To study the effects of verapamil (VR) on proliferation, DNA synthesis and migration of human hepatoma cell line HepG2.

Ecthyma Gangrenosum and Multiple Nodules: Cutaneous Manifestations of Pseudomonas Aeruginosa Sepsis in a Previously Healthy Infant

Pediatric Dermatology. Mar-Apr, 2011  |  Pubmed ID: 21504455

Pseudomonas aeruginosa septicemia is rare in healthy children. Dermatologic manifestations, such as ecthyma gangrenosum and indurated erythematous nodular lesions, as the first signs of pseudomonas infection have rarely been reported. Herein we report a previously healthy 7-month-old girl with ecthyma gangrenosum and multiple nodules as the manifestations of P. aeruginosa sepsis without other systemic involvement.

MicroRNAs Dynamically Remodel Gastrointestinal Smooth Muscle Cells

PloS One. 2011  |  Pubmed ID: 21533178

Smooth muscle cells (SMCs) express a unique set of microRNAs (miRNAs) which regulate and maintain the differentiation state of SMCs. The goal of this study was to investigate the role of miRNAs during the development of gastrointestinal (GI) SMCs in a transgenic animal model. We generated SMC-specific Dicer null animals that express the reporter, green fluorescence protein, in a SMC-specific manner. SMC-specific knockout of Dicer prevented SMC miRNA biogenesis, causing dramatic changes in phenotype, function, and global gene expression in SMCs: the mutant mice developed severe dilation of the intestinal tract associated with the thinning and destruction of the smooth muscle (SM) layers; contractile motility in the mutant intestine was dramatically decreased; and SM contractile genes and transcriptional regulators were extensively down-regulated in the mutant SMCs. Profiling and bioinformatic analyses showed that SMC phenotype is regulated by a complex network of positive and negative feedback by SMC miRNAs, serum response factor (SRF), and other transcriptional factors. Taken together, our data suggest that SMC miRNAs are required for the development and survival of SMCs in the GI tract.

Diet and Cell Size Both Affect Queen-worker Differentiation Through DNA Methylation in Honey Bees (Apis Mellifera, Apidae)

PloS One. 2011  |  Pubmed ID: 21541319

Young larvae of the honey bee (Apis mellifera) are totipotent; they can become either queens (reproductives) or workers (largely sterile helpers). DNA methylation has been shown to play an important role in this differentiation. In this study, we examine the contributions of diet and cell size to caste differentiation.

In Vitro Screening of Ovarian Tumor Specific Peptides from a Phage Display Peptide Library

Biotechnology Letters. Sep, 2011  |  Pubmed ID: 21544611

To develop more biomarkers for diagnosis and therapy of ovarian cancer, a 12-mer phage display library was used to isolate peptides that bound specifically to the human ovarian tumor cell line SK-OV-3. After five rounds of in vitro screening, the recovery rate of phages showed a 69-fold increase over the first round of washings and a group of phage clones capable of binding to SK-OV-3 cells were obtained. A phage clone named Z1 with high affinity and specificity to SK-OV-3 cells was identified in vitro. More importantly, the synthetic biotin-labeled peptide, ZP1 (=SVSVGMKPSPRP), which corresponded to the sequence of the inserted fragment of Z1, demonstrated a high specificity to SK-OV-3 cells especially when compared to other cell lines (A2780 and 3T3). ZP1 might therefore be a biomarker for targeting drug delivery in ovarian cancer therapy.

Oncogene Addiction in Gliomas: Implications for Molecular Targeted Therapy

Journal of Experimental & Clinical Cancer Research : CR. 2011  |  Pubmed ID: 21575270

Oncogene addiction is a phenomenon that the survival of cancer cells depends on an activated oncogene or inactivation of tumor suppressor gene, and is regarded as the 'Achilles heel' of the successful molecular targeted therapies in cancer. However, the role of oncogene addiction in gliomas has not been elucidated systematically. In this review, we summarize the current experimental and clinical evidence for the concept of oncogene addiction and describe the mechanisms explaining oncogene addiction in gliomas. And the clinical implications for oncogene addiction in molecular targeted therapy are further emphasized. In addition, we discuss future direction for defining complex "oncogene addiction network" through the integrated analysis of multiple platforms in the flow of genetic information in gliomagenesis.

Mechanistic Rationale for Inhibition of Poly(ADP-ribose) Polymerase in ETS Gene Fusion-positive Prostate Cancer

Cancer Cell. May, 2011  |  Pubmed ID: 21575865

Recurrent fusions of ETS genes are considered driving mutations in a diverse array of cancers, including Ewing's sarcoma, acute myeloid leukemia, and prostate cancer. We investigate the mechanisms by which ETS fusions mediate their effects, and find that the product of the predominant ETS gene fusion, TMPRSS2:ERG, interacts in a DNA-independent manner with the enzyme poly (ADP-ribose) polymerase 1 (PARP1) and the catalytic subunit of DNA protein kinase (DNA-PKcs). ETS gene-mediated transcription and cell invasion require PARP1 and DNA-PKcs expression and activity. Importantly, pharmacological inhibition of PARP1 inhibits ETS-positive, but not ETS-negative, prostate cancer xenograft growth. Finally, overexpression of the TMPRSS2:ERG fusion induces DNA damage, which is potentiated by PARP1 inhibition in a manner similar to that of BRCA1/2 deficiency.

Overexpression of Osteopontin Induces Angiogenesis of Endothelial Progenitor Cells Via the Avβ3/PI3K/AKT/eNOS/NO Signaling Pathway in Glioma Cells

European Journal of Cell Biology. Aug, 2011  |  Pubmed ID: 21616556

Angiogenesis, a hallmark of tumor growth, is regulated by various angiogenic factors. Recent studies have shown that osteopontin (OPN) is a secreted, integrin-binding protein that contributes to glioma progression. However, its effect on the angiogenesis of gliomas is not fully understood. To elucidate the role of OPN in the process of glioma angiogenesis, endothelial progenitor cells (EPCs) were treated with conditioned media of human glioma SHG44 cells overexpressing OPN. Here, we identified that OPN secreted by glioma cells accelerated EPCs angiogenesis in vitro, including proliferation, migration, and tube formation. OPN also induced the activation of AKT and endothelial nitric oxide synthase (eNOS) and increased NO production without affecting the expression of VEGF, VEGFR-1, or VEGFR-2. Moreover, the avβ3 antibody, the PI3-K inhibitor LY294002 and the eNOS inhibitor NMA suppressed the OPN-mediated increase in NO production and angiogenesis in EPCs. Taken together, these results demonstrate that OPN directly stimulates angiogenesis via the avβ3/PI3-K/AKT/eNOS/NO signaling pathway and may play an important role in tumorigenesis by enhancing angiogenesis in gliomas.

Inhibition of N-myc Downstream-regulated Gene 2 in Prostatic Carcinoma

Cancer Biology & Therapy. Aug, 2011  |  Pubmed ID: 21623166

To study the expression of N-myc Downstream Regulated Gene-2 (NDRG2) in prostatic carcinoma (PCA) tissue and in different PCA cell lines, and to investigate its clinical and pathological implications, 144 PCA and benign prostatic hyperplasia (BPH) tissue sections were analyzed retrospectively with immunohistochemistry (S-P method). The expression levels of NDRG2 and c-Myc in prostate cell lines were detected through Western blot. The effects of adenovirus-mediated NDRG2 on PC3 cells and PC3 nude mouse xenografts was observed through cell growth curves, tumor growth curves, flow cytometry (FCM), transmission electron microscopy (TEM) and TUNEL staining. The NDRG2 gene was highly expressed in BPH tissues, but not in carcinomatous ones (χ(2)=25.98, p < 0.001). Furthermore, positive expression of NDRG2 was negatively correlated with the Gleason score (r = -0.445, p< 0.001) and the c-myc level (r = -0.311, p < 0.001). However, positive expression of NDRG2 was not correlated with pTNM tumor stages or the serum concentration of prostate-specific antigen (PSA) (p > 0.05). The expression of the NDRG2 genes was low in the three PCA cell lines. PC3 cells infected by pAD-cmv-NDRG2 showed inhibition of proliferation both in vitro and vivo. To sum up, NDRG2 may be involved in the carcinogenesis and progression of PCA. Moreover, adenovirus-mediated NDRG2 can suppress the proliferation of PC3 cells significantly both in vitro and in vivo. These results indicate that NDRG2 may become a new target gene for PCA diagnosis and therapy.

[Isolation, Culture and Growth Characteristics of Human Muscle Stem Cells]

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi = Chinese Journal of Otorhinolaryngology Head and Neck Surgery. Apr, 2011  |  Pubmed ID: 21624252

To establish the methods for purification, culture, and identification of adult human skeletal muscle stem cells in vitro and to explore the biological properties of the cells.

Manipulation of Light with α Transformation Media

Journal of the Optical Society of America. A, Optics, Image Science, and Vision. Jun, 2011  |  Pubmed ID: 21643391

A type of transformation media called α media is proposed by performing a direct transformation to the metric tensor of another kind of media, called seed media. Light rays in an α medium correlate to those in its seed medium through a simple displacement or rotation relation. Three types of commonly encountered anisotropic media are covered by the concept of α media: (1) media of slab shape, having continuous translational symmetry with respect to two Cartesian coordinate components; (2) media of cylindrical shape, having cylindrical rotational symmetry and continuous translational symmetry along the longitudinal direction; (3) media of spherical shape, having spherical rotational symmetry, with two principal axes along the symmetry directions, and with the material parameters in the same sign. Optical properties of such media can be effectively interpreted through recalling the properties of certain isotropic media, i.e., their seed media. Conversely, from simple isotropic media in which light trajectories are well known, one can design α media for manipulating light. Based on this fact, several optical devices, including frequency demultiplexers, beam splitters, focusing lenses, and radiation controllers, are designed and numerically verified. The famed invisibility cloak derived from a conventional coordinate transformation is revisited from the α media perspective.

First Construction of Infectious Clone for Newly Emerging Mutation Porcine Circovirus Type 2 (PCV2) Followed by Comparison with PCV2a and PCV2b Genotypes in Biological Characteristics in Vitro

Virology Journal. 2011  |  Pubmed ID: 21658280

Porcine circovirus type 2 (PCV2), the causative agent of postweaning multisystemic wasting syndrome (PMWS), is a serious economic problem in the swine industry. Different genotypes (PCV2a, PCV2b and PCV2d) of the virus are present in the clinical cases in China, and it is necessary to elucidate the pathogenic difference among different genotypes of PCV2. In this study, four strains of different genotypes were isolated, two were ordinary strains and another two were mutation strains, which there are one and two amino acids elongation in the capsid protein (Cap) of PCV2, respectively. Representative strains of different genotypes of the virus were constructed by infectious molecular clone and biological characterization of the rescued viruses were identified in vitro.

A Novel H(2)O(2) Amperometric Biosensor Based on Gold Nanoparticles/self-doped Polyaniline Nanofibers

Bioelectrochemistry (Amsterdam, Netherlands). Oct, 2011  |  Pubmed ID: 21664881

A new kind of gold nanoparticles/self-doped polyaniline nanofibers (Au/SPAN) with grooves has been prepared for the immobilization of horseradish peroxidase (HRP) on the surface of glassy carbon electrode (GCE). The ratio of gold in the composite nanofibers was up to 64%, which could promote the conductivity and biocompatibility of SPAN and increase the immobilized amount of HRP molecules greatly. The electrode exhibits enhanced electrocatalytic activity in the reduction of H(2)O(2) in the presence of the mediator hydroquinone (HQ). The effects of concentration of HQ, solution pH and the working potential on the current response of the modified electrode toward H(2)O(2) were optimized to obtain the maximal sensitivity. The proposed biosensor exhibited a good linear response in the range from 10 to 2000 μM with a detection limit of 1.6 μM (S/N=3) under the optimum conditions. The response showed Michaelis-Menten behavior at larger H(2)O(2) concentrations, and the apparent Michaelis-Menten constant K(m) was estimated to be 2.21 mM. The detection of H(2)O(2) concentration in real sample showed acceptable accuracy with the traditional potassium permanganate titration.

Endogenous Histones Function As Alarmins in Sterile Inflammatory Liver Injury Through Toll-like Receptor 9 in Mice

Hepatology (Baltimore, Md.). Sep, 2011  |  Pubmed ID: 21721026

Sterile inflammatory insults are known to activate innate immunity and propagate organ damage through the recognition of extracellular damage-associated molecular pattern (DAMP) molecules. Although DAMPs such as endogenous DNA and nuclear high-mobility group box 1 have been shown to be critical in sterile inflammation, the role of nuclear histone proteins has not yet been investigated. We report that endogenous histones function as DAMPs after ischemic injury through the pattern recognition receptor Toll-like receptor (TLR) 9 to initiate inflammation. Using an in vivo model of hepatic ischemia/reperfusion (I/R) injury, we show that levels of circulating histones are significantly higher after I/R, and that histone neutralization significantly protects against injury. Injection of exogenous histones exacerbates I/R injury through cytotoxic effects mediated by TLR9 and MyD88. In addition, histone administration increases TLR9 activation, whereas neither TLR9 nor MyD88 mutant mice respond to exogenous histones. Furthermore, we demonstrate in vitro that extracellular histones enhance DNA-mediated TLR9 activation in immune cells through a direct interaction. Conclusion: These novel findings reveal that histones represent a new class of DAMP molecules and serve as a crucial link between initial damage and activation of innate immunity during sterile inflammation.

[Expression of Radioresistant Genes Survivin and HO-1 in Mesenchymal Stem Cells]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Jun, 2011  |  Pubmed ID: 21729576

This study was aimed to investigate the expression of radioresistant genes survivin and HO-1 in mesenchymal stem cells (MSC). Human bone marrow MSC were isolated and enriched by Fircoll density gradient centrifugation, then identified by flow cytometry. MSC were induced with dexamethasone, insulin, 3-isobutyl-1-methyl-xanthine (IBMX) and indomethacin to differentiate into adipocytes. Then the expression of survivin and HO-1 in MSC was detected by RT-PCR. The results indicated that the expressions of surface antigen CD34 and HLA-DR in MSC in vitro were negative while the expressions of CD44 and CD71 were positive. MSC could be differentiated into adipocytes by inductor. RT-PCR showed the expression of radioresistant genes survivin and HO-1 in MSC. It is concluded that MSC have lower sensitivity to radiation, which may associate with the expression of radioresistant genes survivin and HO-1 in MSC.

Single-walled Carbon Nanotube/pyrenecyclodextrin Nanohybrids for Ultrahighly Sensitive and Selective Detection of P-nitrophenol

Langmuir : the ACS Journal of Surfaces and Colloids. Aug, 2011  |  Pubmed ID: 21732642

Electrochemical detection of p-nitrophenol (P-NP) using a highly sensitive and selective platform based on single-walled carbon nanotube/pyrenecyclodextrin (SWCNT/PyCD) nanohybrids is described for the first time. The electrochemical performance of the SWCNT/PyCD nanohybrid electrode was fully compared with bare glassy carbon, single-SWCNT, single-PyCD, and SWCNT/CD (without pyrene rings) electrodes. Besides the techniques of cyclic voltammetry and chronoamperometric transients, differential pulse voltammetry (DPV) has been used for the detection of P-NP without any interference from o-nitrophenol (O-NP) at the potentials of -0.80 and -0.67 V, respectively. The SWCNT/PyCD nanohybrid electrode is highly sensitive, and it shows an ultrahigh sensitivity of 18.7 μA/μM toward P-NP in contrast to the values reported previously. The detection limit (S/N = 3) of the SWCNT/PyCD nanohybrid electrode toward P-NP is 0.00086 μM (0.12 ppb), which is well below the allowed limit in drinking water, 0.43 μM, given by the U.S. Environmental Protection Agency (EPA). The analytical performance of the SWCNT/PyCD nanohybrid electrode toward P-NP is superior to the existing electrodes.

[Research on the Effects of PIAS3 Expression on the Invasion of Glioma TJ905 Cells]

Zhonghua Wai Ke Za Zhi [Chinese Journal of Surgery]. May, 2011  |  Pubmed ID: 21733403

To investigate the function and possible mechanisms of PIAS3 expression on the invasion of TJ905 cells.

A Novel Molecular Mechanism for Nitrated {alpha}-synuclein-induced Cell Death

Journal of Molecular Cell Biology. Aug, 2011  |  Pubmed ID: 21733982

Although previous studies have demonstrated the involvement of nitrated α-synuclein in neurodegenerative disorders (synucleinopathies), the effects of nitrated α-synuclein and the molecular mechanisms underlying its toxicity are still unclear. In the present study, nitrated α-synuclein with four 3-nitrotyrosines (Tyr(39), Tyr(125), Tyr(133), and Tyr(136)) was obtained non-enzymatically by incubation with nitrite. The nitrated protein existed as a mixture of monomers, dimers, and polymers in solution. The nitrated α-synuclein could induce cell death in a time- and concentration-dependent manner when SH-SY5Y cells (a human neuroblastoma cell line) were incubated with the dimers and polymers. Treatment with anti-integrin α5β1 antibody partially rescued the SH-SY5Y cells from the cell death. Dot blotting and immunoprecipitation revealed that the nitrated protein bound to integrin on the cell membranes. Level of nitric oxide (NO) and calcium-independent inducible NO synthase (iNOS) activity increased during the initial stages of the treatment. The expression of phosphorylated focal adhesion kinase (FAK) decreased in the cells. Subsequently, an increase in caspase 3 activity was observed in SH-SY5Y cells. Our results demonstrate that activation of iNOS and inhibition of FAK may both be responsible for the cell death induced by nitrated α-synuclein. These data suggest that the cytotoxicity of nitrated α-synuclein is mediated via an integrin-iNOS/-FAK signaling pathway, and that the nitration of α-synuclein plays a role in neuronal degeneration.

Control of Messenger RNA Fate by RNA Binding Proteins: an Emphasis on Mammalian Spermatogenesis

Journal of Andrology. Jul, 2011  |  Pubmed ID: 21757510

Post-transcriptional status of messenger RNAs (mRNAs) can be affected by many factors, most of which are RNA-binding proteins (RBPs) that either bind mRNAs in a non-specific manner or through specific motifs, usually located in the 3' untranslated regions (UTRs). RBPs can also be recruited by small non-coding RNAs (sncRNAs), which have been shown to be involved in post-transcriptional regulations and transposon repression e.g. microRNAs (miRNAs) or PIWI-interacting RNAs (piRNAs) as components of the sncRNA effector complex. Non-sncRNA-binding RBPs have much more diverse effects on their target mRNAs. Some can cause degradation of their target transcripts and/or repression of translation, whereas others can stabilize and/or activate translation. Splicing and exporting of transcripts from the nucleus to the cytoplasm is often mediated by sequence-specific RBPs. The mechanisms by which RBPs regulate mRNA transcripts involve manipulating the 3'poly(A) tail, targeting the transcript to polysomes or to other ribonuclear protein particles (RNPs), recruiting regulatory proteins, or competing with other RBPs. Here we briefly review the known mechanisms of post-transcriptional regulation mediated by RBPs, with an emphasis on how these mechanisms might control spermatogenesis in general.

Male Germ Cells Express Abundant Endogenous SiRNAs

Proceedings of the National Academy of Sciences of the United States of America. Aug, 2011  |  Pubmed ID: 21788498

In mammals, endogenous siRNAs (endo-siRNAs) have only been reported in murine oocytes and embryonic stem cells. Here, we show that murine spermatogenic cells express numerous endo-siRNAs, which are likely to be derived from naturally occurring double-stranded RNA (dsRNA) precursors. The biogenesis of these testicular endo-siRNAs is DROSHA independent, but DICER dependent. These male germ cell endo-siRNAs can potentially target hundreds of transcripts or thousands of DNA regions in the genome. Overall, our work has unveiled another hidden layer of regulation imposed by small noncoding RNAs during male germ cell development.

The Relationship Between Neuroticism, Major Depressive Disorder and Comorbid Disorders in Chinese Women

Journal of Affective Disorders. Dec, 2011  |  Pubmed ID: 21824661

The personality trait of neuroticism is a risk factor for major depressive disorder (MDD), but this relationship has not been demonstrated in clinical samples from Asia.

Identification of MMP-9 Specific MicroRNA Expression Profile As Potential Targets of Anti-invasion Therapy in Glioblastoma Multiforme

Brain Research. Sep, 2011  |  Pubmed ID: 21831363

The poor prognosis of glioblastoma multiforme (GBM) is largely attributed to their highly invasive nature and MMP-9 plays a pivotal role in regulating invasiveness of malignant glioma cells. MicroRNAs (miRNAs) are small non-coding RNAs that have been shown to regulate a wide range of biological processes via targeting messenger RNA. Previous reports have shown many oncogenes regulate survival and invasion via targeting MMP-9 in GBM. But no literature indicates that miRNAs regulate glioma cell invasion through targeting MMP-9. Here, we show MMP-9 overexpression conferred a poor prognosis in 163 GBM patients. Furthermore, MMP-9 specific miRNA expression profile (14 positively and 31 negatively correlated miRNAs with MMP-9) was established via miRNA microarrays in 60 GBM samples. Among them, two miRNAs: miR-885-5p and miR-491-5p, were chosen for functional validation for their high positive correlation with MMP-9 expression. And upregulation of miR-885-5p and miR-491-5p were demonstrated to reduce the levels of MMP-9 expression and inhibit cellular invasion in U251 and U87 glioma cells. Furthermore, we found that miR-491-5p suppressed glioma cell invasion via targeting MMP-9 directly. To our knowledge, this is the first study to identify the MMP-9 specific microRNA signature which may provide potential targets for anti-invasion therapy in GBM.

Identifiability of Causal Effects for Binary Variables with Baseline Data Missing Due to Death

Biometrics. Aug, 2011  |  Pubmed ID: 21838813

Summary We discuss identifiability and estimation of causal effects of a treatment in subgroups defined by a covariate that is sometimes missing due to death, which is different from a problem with outcomes censored by death. Frangakis et al. (2007, Biometrics 63, 641-662) proposed an approach for estimating the causal effects under a strong monotonicity (SM) assumption. In this article, we focus on identifiability of the joint distribution of the covariate, treatment and potential outcomes, show sufficient conditions for identifiability, and relax the SM assumption to monotonicity (M) and no-interaction (NI) assumptions. We derive expectation-maximization algorithms for finding the maximum likelihood estimates of parameters of the joint distribution under different assumptions. Further we remove the M and NI assumptions, and prove that signs of the causal effects of a treatment in the subgroups are identifiable, which means that their bounds do not cover zero. We perform simulations and a sensitivity analysis to evaluate our approaches. Finally, we apply the approaches to the National Study on the Costs and Outcomes of Trauma Centers data, which are also analyzed by Frangakis et al. (2007) and Xie and Murphy (2007, Biometrics 63, 655-658).

Layer-by-layer Assembly and Electrochemical Study of a 4-aminothiophenol and Ytterbium(III) Trifluoromethanesulfonate Hydrate Film on a Gold Electrode

The Analyst. Oct, 2011  |  Pubmed ID: 21850287

We report on the layer-by-layer assembly and electrochemical properties of 4-aminothiophenol (P-ATP) and ytterbium(III) trifluoromethanesulfonate hydrate (Yb(OTf)(3)) film supported on a gold surface. The fabricated film was characterised electrochemically using redox couples Fe(CN)(6)(3-/4-), complemented with imaging using atomic force microscopy (AFM). The electrocatalytic activity of the prepared electrodes was studied using cyclic and differential pulse voltammetries. Electrochemical measurements show that the P-ATP/Yb(OTf)(3) modified electrode has superb activity towards hydroquinone (HQ) oxidation and that there is a significant improvement in the electrode stability and reproducibility due to the covalent and coordination reactions.

Identification of One Critical Amino Acid That Determines a Conformational Neutralizing Epitope in the Capsid Protein of Porcine Circovirus Type 2

BMC Microbiology. 2011  |  Pubmed ID: 21859462

Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS) in pigs. Currently, there is considerable interest in the immunology of PCV2; in particular, the immunological properties of the capsid protein. This protein is involved in PCV2 immunogenicity and is a potential target for vaccine development. In this study, we identified one critical amino acid that determines a conformational neutralizing epitope in the capsid protein of PCV2.

[Secreted Expression of Dengue Virus Type 2 Envelope Glycoprotein in Eukaryotic Cells]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Apr, 2011  |  Pubmed ID: 21863624

To secreted express envelope glycoprotein (E) of dengue virus type 2 extracellularly.

Isolated Angiitis of the Central Nervous System with Tumor-like Lesion, Mimicking Brain Malignant Glioma: a Case Report and Review of the Literature

World Journal of Surgical Oncology. 2011  |  Pubmed ID: 21867556

Isolated angiitis of the central nervous system (IACNS) is a rare but severe vascular disease, which could present like an isolated inflammatory lesion on magnetic resonance imaging (MRI). To date, only a few such cases with tumor-like IACNS have been reported.

[Ki-67 Proliferative Index in Non-Hodgkin's Lymphoma and Its Clinical Significance]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Aug, 2011  |  Pubmed ID: 21867618

This study was aimed to investigate the relationship of Ki-67 proliferation index (Ki-67 PI) with non-Hodgkin's lymphoma(NHL) typing and biological behavior, as well as its significance in clinical characters and prognosis of diffuse large B-cell lymphoma(DLBCL). A total of 542 cases of NHL in our hospital from 1st January 2001 to 31st December 2010 were retrospectively analyzed, and Ki-67 PI was all assayed immunohistochemically, and a total of 82 cases of newly-diagnosed DLBCL with more clinical records were investigated. The results indicated that according to the World Health Organization (WHO) histopathological classification of lymphoma, Ki-67 PI was different as classification for NHL subgroups was different. The Ki-67 PI increased with aggressive progression of NHL. The mean Ki-67 PI ranged from 25.5% in indolent lymphoma to 98.4% in very aggressive lymphoma. ROC curve analysis demonstrated that the 50% was the cut-off value distinguishing indolent from aggressive disease. On ROC curve analysis, Ki-67 PI of 75% was found to significantly discriminate patients with DLBCL who had a good or bad prognosis. There was a significant correlation of Ki-67 PI with Ann Arbor stage and LDH level. When the DLBCL cases were divided by Ann Arbor stage and IPI score, the 3-year overall survival (OS) of patients with a low Ki-67 PI (≤ 75%) in the group of Ann Arbor stage III-IV and high LDH level was higher than those with a high Ki-67 PI (> 75%) among the patients with B symptoms and IPI 3.0-5. 3-year OS in those with a low Ki-67 PI (≤ 75%) in the group of Ann Arbor stage III-IV and normal LDH level was higher than those with a high Ki-67 PI (> 75%) among the patients with B symptoms. 3-year OS of patients with a low Ki-67 PI (≤ 75%) in the group at III-IV stage and a high LDH level was higher than those with a high Ki-67 PI (> 75%). It is concluded that a cut-off value of 50% can be helpful to differentiate indolent from aggressive NHL. In DLBCL, a cut-off value of 75% can distinguish patients with a good or bad prognosis when combined with other prognostic factors, i.e. B symptoms, Ann Arbor stage, IPI score and lactate dehydrogenase (LDH) level.

Inhibition of STAT3 Reverses Alkylator Resistance Through Modulation of the AKT and β-catenin Signaling Pathways

Oncology Reports. Nov, 2011  |  Pubmed ID: 21887474

Activation of signal transducer and activator of transcription 3 (STAT3) is associated with poor clinical outcome of glioblastoma (GBM). However, the role of STAT3 in resistance to alkylator-based chemotherapy remains unknown. Here, we retrospectively analyzed the phosphorylated STAT3 (p-STAT3) profile of 68 GBM patients receiving alkylator therapy, identifying p-STAT3 as an independent unfavorable prognostic factor for progression-free and overall survival. Additionally, elevated p-STAT3 expression correlated with resistance to alkylator therapy. In vitro analysis revealed that U251 and U87 human glioma cells were refractory to treatment with the common alkylating agent temozolomide (TMZ), with only a modest impact on AKT and β-catenin activation in the context of high p-STAT3. Inhibition of STAT3 in these cells significantly enhanced the effect of TMZ. Inhibition of STAT3 dramatically decreased the IC50 of TMZ, increasing TMZ-induced apoptosis while up-regulating expression of Bcl-2 and down-regulating expression of Bax. Furthermore, inhibition of STAT3 increased TMZ-induced G₀-G₁ arrest and decreased Cyclin D1 expression compared to TMZ alone. Together, these results indicate that inhibition of STAT3 sensitizes glioma cells to TMZ, at least in part, by blocking the p-AKT and β-catenin pathways. These findings strongly support the hypothesis that STAT3 inhibition significantly improves the clinical efficacy of alkylating agents.

Magnetic Biodegradable Fe3O4/CS/PVA Nanofibrous Membranes for Bone Regeneration

Biomedical Materials (Bristol, England). Oct, 2011  |  Pubmed ID: 21893702

In recent years, interest in magnetic biomimetic scaffolds for tissue engineering has increased considerably. The aim of this study is to develop magnetic biodegradable fibrous materials with potential use in bone regeneration. Magnetic biodegradable Fe(3)O(4)/chitosan (CS)/poly vinyl alcohol (PVA) nanofibrous membranes were achieved by electrospinning with average fiber diameters ranging from 230 to 380 nm and porosity of 83.9-85.1%. The influences of polymer concentration, applied voltage and Fe(3)O(4) nanoparticles loading on the fabrication of nanofibers were investigated. The polymer concentration of 4.5 wt%, applied voltage of 20 kV and Fe(3)O(4) nanoparticles loading of lower than 5 wt% could produce homogeneous, smooth and continuous Fe(3)O(4)/CS/PVA nanofibrous membranes. X-ray diffraction (XRD) data confirmed that the crystalline structure of the Fe(3)O(4), CS and PVA were maintained during electrospinning process. Fourier transform infrared spectroscopy (FT-IR) demonstrated that the Fe(3)O(4) loading up to 5 wt% did not change the functional groups of CS/PVA greatly. Transmission electron microscopy (TEM) showed islets of Fe(3)O(4) nanoparticles evenly distributed in the fibers. Weak ferrimagnetic behaviors of membranes were revealed by vibrating sample magnetometer (VSM) test. Tensile test exhibited Young's modulus of membranes that were gradually enhanced with the increase of Fe(3)O(4) nanoparticles loading, while ultimate tensile stress and ultimate strain were slightly reduced by Fe(3)O(4) nanoparticles loading of 5%. Additionally, MG63 human osteoblast-like cells were seeded on the magnetic nanofibrous membranes to evaluate their bone biocompatibility. Cell growth dynamics according to MTT assay and scanning electron microscopy (SEM) observation exhibited good cell adhesion and proliferation, suggesting that this magnetic biodegradable Fe(3)O(4)/CS/PVA nanofibrous membranes can be one of promising biomaterials for facilitation of osteogenesis.

High Adsorptive γ-AlOOH(boehmite)@SiO2/Fe3O4 Porous Magnetic Microspheres for Detection of Toxic Metal Ions in Drinking Water

Chemical Communications (Cambridge, England). Oct, 2011  |  Pubmed ID: 21897953

γ-AlOOH(boehmite)@SiO(2)/Fe(3)O(4) porous magnetic microspheres with high adsorption capacity toward heavy metal ions were found to be useful for the simultaneous and selective electrochemical detection of five metal ions, such as ultratrace zinc(II), cadmium(II), lead(II), copper(II), and mercury(II), in drinking water.

Variable Volume Loading Method: a Convenient and Rapid Method for Measuring the Initial Emittable Concentration and Partition Coefficient of Formaldehyde and Other Aldehydes in Building Materials

Environmental Science & Technology. Dec, 2011  |  Pubmed ID: 21939215

The initial emittable formaldehyde and VOC concentration in building materials (C(0)) is a key parameter for characterizing and classifying these materials. Various methods have been developed to measure this parameter, but these generally require a long test time. In this paper we develop a convenient and rapid method, the variable volume loading (VVL) method, to simultaneously measure C(0) and the material/air partition coefficient (K). This method has the following features: (a) it requires a relatively short experimental time (less than 24 h for the cases studied); and (b) is convenient for routine measurement. Using this method, we determined C(0) and K of formaldehyde, propanal and hexanal in one kind of medium density fiberboard, and repeated experiments were performed to reduce measurement error. In addition, an extended-C-history method is proposed to determine the diffusion coefficient and the convective mass transfer coefficient. The VVL method is validated by comparing model predicted results based on the determined parameters with experimental data. The determined C(0) of formaldehyde obtained via this method is less than 10% of the total concentration using the perforator method recommended by the Chinese National Standard, suggesting that the total concentration may not be appropriate to predict emission characteristics, nor for material classification.

New Candidate Tumor-Suppressor Gene KLF6 and Its Splice Variant KLF6 SV2 Counterbalancing Expression in Primary Hepatocarcinoma

Hepato-gastroenterology. Aug, 2011  |  Pubmed ID: 21940380

Background/Aims: This study aimed to detect the expression of newly discovered zinc finger transcriptional factor KLF6 and its splice variant KLF6 SV2 in primary hepatocarcinoma (PHC) tissues and hepatoma cell strains, and to evaluate their clinicopathologic relationship with PHC. Methodology: Wild-type KLF6 and KLF6 SV2 mRNA expression was determined by RT-PCR in 27 cases of PHC tissues and cell strains of HepG2, SMMC7721 and LO2. Western blotting and immunohistochemical staining were adopted to detect KLF6 protein expression. Positive area ratio of wild-type KLF6 protein expression and its relationship with clinicopathological parameters of PHC was analyzed. Results: Wild-type KLF6 expression in PHC tissues was lower than that in paracancerous tissues. In contrast, KLF6 SV2 mRNA expression was higher in PHC tissues and hepatoma cell strains (p<0.05). Positive area ratio of wild-type KLF6 protein expression was positively correlated with cellular differentiation degree of PHC (p<0.01), but negatively correlated not only with liver cirrhosis, tumor size and extrahepatic metastases (p<0.01), but also with portal vein thrombus and the number of lymph nodes with metastasis (p<0.05). Conclusions: Wild-type KLF6 deletion and inactivation was involved in the growth, cell differentiation and other physiological processes of PHC. The upregulation of KLF6 splice variant might counterbalance the wild-type KLF6 and contribute to the occurrence and development of PHC.

D-ribose Induces Cellular Protein Glycation and Impairs Mouse Spatial Cognition

PloS One. 2011  |  Pubmed ID: 21966363

D-ribose, an important reducing monosaccharide, is highly active in the glycation of proteins, and results in the rapid production of advanced glycation end products (AGEs) in vitro. However, whether D-ribose participates in glycation and leads to production of AGEs in vivo still requires investigation.

Gossypin Induces G2/M Arrest in Human Malignant Glioma U251 Cells by the Activation of Chk1/Cdc25C Pathway

Cellular and Molecular Neurobiology. Oct, 2011  |  Pubmed ID: 21984341

Gossypin is a flavone that was originally isolated from Hibiscus vitifolius and has traditionally been used for the treatment of diabetes, jaundice, and inflammation. Recently, gossypin was found to have potent anticancer properties; however, its effect on human gliomas still remain unknown. To investigate the potential anticancer effects of gossypin on malignant gliomas and analyze the associated molecular mechanisms, we treated human glioma U251 cells with gossypin. Our study showed that the treatment of U251 cells with gossypin inhibited cell proliferation in a dose- and time-dependent manner and was observed to be minimally toxic to normal human astrocytes. Gossypin's effect on cell cycle distribution was observed, and we found that it induced G2/M-phase arrest in U251 cells. An analysis of cell-cycle regulatory proteins indicated that the arresting effect of gossypin on the cell cycle at G2/M phase was involved in the phosphorylation of cell division cycle 25C (Cdc25C) tyrosine phosphatase via the activation of checkpoint kinase 1 (Chk1). These findings indicate that gossypin is a potential treatment of gliomas because of gossypin's potential to regulate the proliferation of U251 cells via the cell-cycle regulatory proteins Chk1 and Cdc25C.

Inhibitory Activity of YKL-40 in Mammary Epithelial Cell Differentiation and Polarization Induced by Lactogenic Hormones: a Role in Mammary Tissue Involution

PloS One. 2011  |  Pubmed ID: 21991364

We previously reported that a secreted glycoprotein YKL-40 acts as an angiogenic factor to promote breast cancer angiogenesis. However, its functional role in normal mammary gland development is poorly understood. Here we investigated its biophysiological activity in mammary epithelial development and mammary tissue morphogenesis. YKL-40 was expressed exclusively by ductal epithelial cells of parous and non-parous mammary tissue, but was dramatically up-regulated at the beginning of involution. To mimic ductal development and explore activity of elevated YKL-40 during mammary tissue regression in vivo, we grew a mammary epithelial cell line 76N MECs in a 3-D Matrigel system in the presence of lactogenic hormones including prolactin, hydrocortisone, and insulin. Treatment of 76N MECs with recombinant YKL-40 significantly inhibited acinar formation, luminal polarization, and secretion. YKL-40 also suppressed expression of E-cadherin but increased MMP-9 and cell motility, the crucial mechanisms that mediate mammary tissue remodeling during involution. In addition, engineering of 76N MECs with YKL-40 gene to express ectopic YKL-40 recapitulated the same activities as recombinant YKL-40 in the inhibition of cell differentiation. These results suggest that YKL-40-mediated inhibition of cell differentiation and polarization in the presence of lactogenic hormones may represent its important function during mammary tissue involution. Identification of this biophysiological property will enhance our understanding of its pathologic role in the later stage of breast cancer that is developed from poorly differentiated and highly invasive cells.

Gene Expression Profiling Reveals Ki-67 Associated Proliferation Signature in Human Glioblastoma

Chinese Medical Journal. Sep, 2011  |  Pubmed ID: 22040407

Everlasting cellular proliferation is the fundamental feature during gliomagenesis and Ki-67 is one of the classical proliferation markers in human glioblastoma multiforme (GBM). However, the driver genes or core pathways for cellular proliferation in GBM have not been elucidated systematically.

Relationship Between Magnetic Resonance Imaging and Molecular Pathology in Patients with Glioblastoma Multiforme

Chinese Medical Journal. Sep, 2011  |  Pubmed ID: 22040408

Glioblastoma multiforme (GBM) is the most common and lethal primary brain tumor in adults. Magnetic resonance imaging (MRI) is routinely used in the diagnosis, characterization and clinical management of GBM. The diagnosis and treatment of GBM is largely guided by histopathology and immunohistochemistry. This study aimed to identify the relationship between magnetic resonance features and molecular pathology of GBM.

[Biological Effects of Low Dose X-irradiation on Human Bone Marrow Mesenchymal Stem Cells]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Oct, 2011  |  Pubmed ID: 22040974

Recent studies have shown that low dose X-irradiation shows specific effect different from high dose exposures. However, the biologic responses of bone marrow mesenchymal stem cells (BM-MSC) to low dose X-irradiation have rarely been described in the literature. This study was purposed to investigate the biologic responses of human bone marrow-derived MSC to low dose X-irradiation. The proliferation of cells was tested with MTT assay, so that the cell growth curve was drawn at 1 to 7 days. The expression of survivin mRNA was detected by RT-PCR assay; the BM-MSC DNA damage induced by X-irradiation were detected with mononuclear cell gel electrophoresis. The results indicated that the proliferative ability of BM-MSC exposed to low doses of X rays was obviously enhanced as compared with control group. The low dose X-irradiation caused the damage of DNA in X-ray dose dependent manner. X-irradiation enhanced expression of survivin in MSC. It is concluded that the low dose below 20 cGy of X-irradiation has a promoting effect on suvivin expression in BM-MSC. Whether the high expression of survivin plays an important role to resist ionizing radiation needs to be further studied.

Berberine and Itraconazole Are Not Synergistic in Vitro Against Aspergillus Fumigatus Isolated from Clinical Patients

Molecules (Basel, Switzerland). 2011  |  Pubmed ID: 22051933

The incidence of Aspergillus fumigatus infections has become more frequent as a consequence of widespread immunosuppression. At present, the number of available antifungal agents in the clinic is limited, and most of them, such as itraconazole (ICZ), are toxic and show resistance. Berberine (BER) is a plant alkaloid used in the clinic mainly for alimentary infections. We have used BER and ICZ to measure in vitro resistance in A. fumigatus isolated from clinical patients. The minimum inhibitory concentration ranges of BER and ICZ were 4-256 and 0.031-0.250 μg/mL, respectively. In addition, against A. fumigatus IFM 40808 strain, the MIC₅₀ values of BER and ICZ were 8 and 0.125 μg/mL. Using this strain, we compared the giant colonies with or without BER, and concluded that BER could restrain A. fumigatus mycelial growth and conidial pigment production. Combinations of the two drugs were also tested by the checkerboard assay to identify any functional interactions between them. Thirty-two out of 42 isolates had FICI values > 4.0, indicating that two drugs were mutually antagonistic. In conclusion, it is not advised that BER and ICZ be used in the clinic at the same time. Our results indicated that BER may inhibit A. fumigatus through the ergosterol biosynthesis pathway, like ICZ.

Structural Basis of γH2AX Recognition by Human PTIP BRCT5-BRCT6 Domains in the DNA Damage Response Pathway

FEBS Letters. Dec, 2011  |  Pubmed ID: 22064073

Human Pax2 transactivation domain-interacting protein (hPTIP), containing six BRCT domains, is an essential protein required for the IR induced DDR process with an unclear role. Here we report that the tandem BRCT5-BRCT6 domain of hPTIP recognizes the γH2AX tail, and this interaction depends on the phosphorylation of H2AX Ser139 and binding with the carboxyl ending peptide to the aminoacyl ending peptide. The 2.15 Å crystal structure of hPTIP BRCT5/6-γH2AX complex and mutation analysis provide molecular evidence for direct interactions between PTIP and γH2AX. This interaction proffers a new clue to identify the role of PTIP in DDR pathways.

The Population and Evolutionary Dynamics of Vibrio Cholerae and Its Bacteriophage: Conditions for Maintaining Phage-limited Communities

The American Naturalist. Dec, 2011  |  Pubmed ID: 22089867

Although bacteriophage have been reported to be the most abundant organisms on earth, little is known about their contribution to the ecology of natural communities of their host bacteria. Most importantly, what role do these viral parasitoids play in regulating the densities of bacterial populations? To address this question, we use experimental communities of Vibrio cholerae and its phage in continuous culture, and we use mathematical models to explore the population dynamic and evolutionary conditions under which phage, rather than resources, will limit the densities of these bacteria. The results of our experiments indicate that single species of bacterial viruses cannot maintain the density of V. cholerae populations at levels much lower than that anticipated on the basis of resources alone. On the other hand, as few as two species of phage can maintain these bacteria at densities more than two orders of magnitude lower than the densities of the corresponding phage-free controls for extensive periods. Using mathematical models and short-term experiments, we explore the population dynamic processes responsible for these results. We discuss the implications of this experimental and theoretical study for the population and evolutionary dynamics of natural populations of bacteria and phage.

Identification of Estrogen-associated Intrinsic Aging Genes in Chinese Han Female Skin by CDNA Microarray Technology

Biomedical and Environmental Sciences : BES. Aug, 2011  |  Pubmed ID: 22108325

Estrogens play an important role in intrinsic skin aging. The associated changes in global gene expression are poorly understood.

Construction of Porcine Growth Hormone Eukaryotic Expression Vector and Its Transfection Mediated by Cationic Liposome in Mice

Animal Biotechnology. Oct, 2011  |  Pubmed ID: 22132815

The present study was designed to construct the eukaryotic expression vector for pGH mature peptide (mpGH) and to investigate its transfection mediated by cationic liposome (CLs) in COS-7 cells and mice. The cDNA of mpGH ORF was successfully cloned by reverse transcription-PCR (RT-PCR) using the adult pig pituitary gland RNA. The recombinant eukaryotic expression vector, VmpGH, was constructed by ligating the cDNA fragment to the vector VR1020. The successful construction was confirmed by restriction enzyme digestion, and the expression of mpGH was confirmed by RT-PCR, immunofluorescence analyses (IFA), and ELISA in COS-7 cells. The VmpGH and VR1020 plasmids were entrapped with CLs, and four experimental groups of male Kunming mice were administrated with VmpGH / lipoplex or naked VmpGH plasmids at two dosages (0.5 and 1.0 mg/kg), while the mice injected with VR1020-lipoplex at the dosage of 0.5 mg/kg body weight (BW) were used as control. The BWs of the mice administrated with VmpGH-lipoplex at both dosages were significantly higher than not only those of the control (P < 0.01) but also those of mice injected with naked plasmids (P < 0.01), from 30 to 60 days post-transfection. The transcription of VmpGH was detected by RT-PCR in six tissues, including the liver, kidney, spleen, heart, muscle, and blood, of the mice injected with VmpGH-lipoplex, but not in the same tissues of control mice. Furthermore, the mice injected with VmpGH-lipoplex showed higher plasma GH contents than the control mice (P < 0.05), although their IgG contents did not show much difference. Our study demonstrates that the VmpGH plasmids' transfection mediated by CLs can significantly promote the growth of mice, which may be used to improve the livestock production.

Specific Recognition of Phosphorylated Tail of H2AX by the Tandem BRCT Domains of MCPH1 Revealed by Complex Structure

Journal of Structural Biology. Dec, 2011  |  Pubmed ID: 22154951

MCPH1 is especially important for linking chromatin remodeling to DNA damage response. It contains three BRCT (BRCA1-carboxyl terminal) domains. The N-terminal region directly binds with chromatin remodeling complex SWI-SNF, and the C-terminal BRCT2-BRCT3 domains (tandem BRCT domains) are involved in cellular DNA damage response. The MCPH1 gene associates with evolution of brain size, and its variation can cause primary microcephaly. In this study we solve the crystal structures of MCPH1 natural variant (A761) C-terminal tandem BRCT domains alone as well as in complex with γH2AX tail. Compared with other structures of tandem BRCT domains, the most significant differences lie in phosphopeptide binding pocket. Additionally, fluorescence polarization assays demonstrate that MCPH1 tandem BRCT domains show a binding selectivity on pSer +3 and prefer to bind phosphopeptide with free COOH-terminus. Taken together, our research provides new structural insights into BRCT-phosphopeptide recognition mechanism.

Metalloproteinase-mediated Shedding of Integrin β2 Promotes Macrophage Efflux from Inflammatory Sites

The Journal of Biological Chemistry. Feb, 2012  |  Pubmed ID: 22170060

Macrophage exiting from inflammatory sites is critical to limit the local innate immune response. With tissue insult, resident tissue macrophages rapidly efflux to lymph nodes where they modulate the adaptive immune response, and inflammatory macrophages attracted to the site of injury then exit during the resolution phase. However, the mechanisms that regulate macrophage efflux are poorly understood. This study has investigated soluble forms of integrin β2 whose levels are elevated in experimental peritonitis at times when macrophages are exiting the peritoneum, suggesting that its proteolytic shedding may be involved in macrophage efflux. Both constitutive and inducible metalloproteinase-dependent shedding of integrin β2 from mouse macrophages are demonstrated. Soluble integrin β2 is primarily released as a heterodimeric complex with αM that retains its ability to bind its ligands intracellular adhesion molecule-1, fibrin, and collagen and thus may serve as a soluble antagonist. In a model of accelerated exiting, administration of a metalloproteinase inhibitor prevents macrophage efflux by 50% and impedes loss of macrophage integrin β2 from the cell surface. Exiting of peritoneal macrophages in mice lacking integrin β2 is accelerated, and antibody disruption of integrin β2-substrate interactions can reverse 50% of the metalloprotease inhibitor blockade of macrophage exiting. Thus, our study demonstrates the ability of metalloproteinase-mediated shedding of integrin β2 to promote macrophage efflux from inflammatory sites, and the release of soluble integrin heterodimers may also limit local inflammation.

Interferon-gamma Induces Autophagy with Growth Inhibition and Cell Death in Human Hepatocellular Carcinoma (HCC) Cells Through Interferon-regulatory Factor-1 (IRF-1)

Cancer Letters. Jan, 2012  |  Pubmed ID: 22056812

Interferon-gamma (IFN-γ) is a pleiotropic cytokine with immunomodulatory, anti-viral, and anti-proliferative effects. In this study, we examined the effects of IFN-γ on autophagy and cell growth in human hepatocellular carcinoma (HCC) cells. IFN-γ inhibited cell growth of Huh7 cells with non-apoptotic cell death. IFN-γ induced autophagosome formation and conversion/turnover of microtubule associated protein 1 light chain 3 (LC3) protein. Furthermore, overexpression of IRF-1 also induced autophagy in Huh7 cells. Silencing IRF-1 expression with target small hairpin RNA blocked autophagy induced by IFN-γ. Silencing of the autophagy signals Beclin-1 or Atg5 attenuated the inhibitory effect of IFN-γ on Huh7 cells with decreased cell death. Additionally, IFN-γ activated autophagy in freshly cultured human HCC cells. Together, these findings show that IFN-γ induces autophagy through IRF-1 signaling pathway and the induction of autophagy contributes to the growth-inhibitory effect of IFN-γ with cell death in human liver cancer cells.

Formation Mechanism of Monodisperse, Low Molecular Weight Chitosan Nanoparticles by Ionic Gelation Technique

Colloids and Surfaces. B, Biointerfaces. Feb, 2012  |  Pubmed ID: 22014934

Chitosan nanoparticles have been extensively studied for drug and gene delivery. In this paper, monodisperse, low molecular weight (LMW) chitosan nanoparticles were prepared by a novel method based on ionic gelation using sodium tripolyphosphate (TPP) as cross-linking agent. The objective of this study was to solve the problem of preparation of chitosan/TPP nanoparticles with high degree of monodispersity and stability, and investigate the effect of various parameters on the formation of LMW chitosan/TPP nanoparticles. It was found that the particle size distribution of the nanoparticles could be significantly narrowed by a combination of decreasing the concentration of acetic acid and reducing the ambient temperature during cross-linking process. The optimized nanoparticles exhibited a mean hydrodynamic diameter of 138 nm with a polydispersity index (PDI) of 0.026 and a zeta potential of +35 mV, the nanoparticles had good storage stability at room temperature up to at least 20 days.

MicroRNA-125b-2 Confers Human Glioblastoma Stem Cells Resistance to Temozolomide Through the Mitochondrial Pathway of Apoptosis

International Journal of Oncology. Jan, 2012  |  Pubmed ID: 21879257

MicroRNAs (miRNAs) are small, non-coding RNA molecules that regulate protein expression by cleaving or repressing the translation of target mRNAs. miR-125b, one of the neuronal miRNAs, was recently found to be necessary for stem cell fission and for making stem cells insensitive to chemotherapy signals. Temozolomide (TMZ) is a promising chemotherapeutic agent for treating glioblastomas. However, resistance develops quickly and with a high frequency. Given the insensitivity of some glioblastomas to TMZ and the hypothesis that glioma stem cells cause resistance to drug therapy, exploring the functions and mechanisms of miR-125b action on TMZ-treated glioblastoma stem cells would be valuable. In this study, we found that miR-125b-2 is overexpressed in glioblastoma multiforme tissues and the corresponding stem cells (GBMSC); downregulation of miR-125b-2 expression in GBMSC could allow TMZ to induce GBMSC apoptosis. Additionally, the expression of the anti-apoptotic protein Bcl-2 was decreased after the TMZ+miR-125b-2 inhibitor treatment, while the expression of the proapoptotic protein Bax was increased. Further research demonstrated that the induction of apoptosis in GBMSC is also associated with increased cytochrome c release from mitochondria, induction of Apaf-1, activation of caspase-3 and poly-ADP-ribose polymerase (PARP). Taken together, these results suggest that miR-125b-2 overexpression might confer glioblastoma stem cells resistance to TMZ.

A Prospective, Randomized, Multicenter Acceptability and Safety Study of Direct Buprenorphine/naloxone Induction in Heroin-dependent Individuals

Addiction (Abingdon, England). Jan, 2012  |  Pubmed ID: 21749526

To provide controlled data on direct induction with buprenorphine/naloxone (BNX) versus indirect buprenorphine (BPN)-to-BNX induction.

The Novel Lupus Antigen Related Protein Acheron Enhances the Development of Human Breast Cancer

International Journal of Cancer. Journal International Du Cancer. Feb, 2012  |  Pubmed ID: 21387291

Acheron (Achn) is a new member of the Lupus antigen family of RNA binding proteins. Previous studies have shown that Achn controls developmental decisions in neurons and muscle. In the human mammary gland, Achn expression is restricted to ductal myoepithelial cells. Microarray analysis and immunohistochemistry have shown that Achn expression is elevated in some basal-like ductal carcinomas. To study the possible role of Achn in breast cancer, we engineered human MDA-MB-231 cells to stably express enhanced green fluorescent protein-tagged wild-type Achn (AchnWT), as well as Achn lacking either its nuclear localization signal (AchnNLS) or its nuclear export signal (AchnNES). In in vitro assays, AchnWT and AchnNES, but not AchnNLS, enhanced cell proliferation, lamellipodia formation, and invasive activity and drove expression of the elevated expression of the metastasis-associated proteins MMP-9 and VEGF. To determine if Achn could alter the behavior of human breast cancer cells in vivo, Achn-engineered MDA-MB-231 cells were injected into athymic SCID/Beige mice. AchnWT and AchnNES-expressing tumors displayed enhanced angiogenesis and an approximately 5-fold increase in tumor size relative to either control cells or those expressing AchnNLS. These data suggest that Achn enhances human breast tumor growth and vascularization and that this activity is dependent on nuclear localization.

Spatial Learning and Memory Impairment and Pathological Change in Rats Induced by Acute Exposure to Microcystin-LR

Environmental Toxicology. Jan, 2012  |  Pubmed ID: 22223477

Microcystin-LR (MCLR) is a commonly encountered blue-green algal hepatotoxin and a known inhibitor of cellular protein phosphatase. However, little is known about its neurotoxicity. By using Morris water maze, histopathological and biochemical analysis, we investigated MCLR-induced neurotoxicity on the hippocampus of rat brain. After rats were intrahippocampally injected with MCLR (1 and 10 μg/L), their learning and memory function was greatly impaired, suggesting the neurotoxic potential of MCLR. Meanwhile, obvious histological and ultrastructural injuries and serious oxidative damage were also observed in the hippocampus. These results suggested that oxidative stress might be involved in the MCLR-induced pathological damage in hippocampus, subsequently leading to the spatial learning and memory deficit of rat. Taken together, our results highlighted the MCLR-induced neurotoxicity in the rat, as well as the importance of oxidative stress and pathological impairment in this procedure. © 2011 Wiley Periodicals, Inc. Environ Toxicol, 2011.

High Mobility Group Box 1 Activates Caspase-1 and Promotes Hepatocellular Carcinoma Invasiveness and Metastases

Hepatology (Baltimore, Md.). Jan, 2012  |  Pubmed ID: 22234969

Hypoxia is often found in solid tumors and is associated with tumor progression and poor clinical outcomes. The exact mechanisms related to hypoxia-induced invasion and metastasis remain unclear. We elucidated the mechanism by which the nuclear damage associated molecular pattern molecule, high mobility group box 1 (HMGB1), released under hypoxic stress can induce an inflammatory response to promote invasion and metastasis in hepatocellular carcinoma (HCC) cells. Caspase-1 activation was found to occur in hypoxic HCC cells in a process that was dependent on the extracellular release of HMGB1 and subsequent activation of both TLR4 and RAGE signaling pathways. Downstream from hypoxia induced caspase-1 activation, cleavage and release of proinflammatory cytokines, IL-1β and IL-18 occurred. We further demonstrate that overexpression of HMGB1 or treatment with recombinant HMGB1 enhanced invasiveness of HCC cells while stable knockdown of HMGB1 remarkably reduced HCC invasion. Moreover, in a murine model of HCC pulmonary metastasis, stable knockdown of HMGB1 suppressed HCC invasion and metastasis. Conclusion: These results suggest that in hypoxic HCC cells, HMGB1 activates TLR4 and RAGE signaling pathways to induce caspase-1 activation with subsequent production of multiple inflammatory mediators which in turn promotes cancer invasion and metastasis. (HEPATOLOGY 2012.).

Resemblance of Symptoms for Major Depression Assessed at Interview Versus from Hospital Record Review

PloS One. 2012  |  Pubmed ID: 22247760

Diagnostic information for psychiatric research often depends on both clinical interviews and medical records. Although discrepancies between these two sources are well known, there have been few studies into the degree and origins of inconsistencies.

Splenocyte Apoptosis and Autophagy is Mediated by Interferon Regulatory Factor-1 During Murine Endotoxemia

Shock (Augusta, Ga.). Jan, 2012  |  Pubmed ID: 22266972

ABSTRACT: Sepsis induced lymphocyte and dendritic cell apoptosis contributes to immunosuppression, which results in an inability to eradicate the primary infection as well as a propensity to acquire new, secondary infections. Another cellular process, autophagy, is also activated in immune cells and plays a protective role. In the present study, we demonstrate that interferon regulatory factor-1 (IRF-1) regulates both immune cell apoptosis and autophagy in a murine endotoxemia model. IRF-1 is activated at an early phase through a Toll-like receptor 4 (TLR4)-dependent, myeloid differentiation primary response gene 88 (MyD88)-independent manner in splenocytes. Further, IRF-1 knockout (KO) mice are protected from a lethal endotoxemia model. This protection is associated with decreased apoptosis and increased autophagy in splenocytes. IRF-1 KO mice experience decreased apoptotic cell loss, especially in CD4+ T lymphocytes and myeloid antigen presenting cells (APC). Meanwhile, IRF-1 KO mice demonstrate increased autophagy and improved mitochondrial integrity. This increased autophagy in KO mice is attributable, at least in part, to deactivation of mammalian target of rapamycin (mTOR)/P70S6 signaling - a main negative regulator of autophagy. Therefore, we propose a novel role for IRF-1 in regulating both apoptosis and autophagy in splenocytes in the setting of endotoxemia with IRF-1 promoting apoptosis and inhibiting autophagy.

D-ribose in Glycation and Protein Aggregation

Biochimica Et Biophysica Acta. Jan, 2012  |  Pubmed ID: 22274132

BACKGROUND: d-ribose is a naturally occurring pentose monosaccharide present in all living cells and their microenvironments and is a key component of numerous biomolecules involved in many important metabolic pathways. It also participates in the glycation of proteins producing advanced glycation end products (AGEs) that lead to cell dysfunction and death. As recent studies show, ribosylation, a rapid process, causes protein aggregation in vitro and in vivo. SCOPE OF REVIEW: This review summarizes the relationship between ribosylation, protein aggregation, cell death and cognitive impairments. MAJOR CONCLUSION: d-ribose is active in glycation and induces protein aggregation, rapidly producing AGEs in vitro and in vivo. GENERAL SIGNIFICANCE: Ribosylation, leading to the production of significant amounts of AGEs both extracellularly and intracellularly, may be involved in cell dysfunction and subsequent cognitive impairments. This review may be a useful reference for studies on the pharmacokinetics of d-ribose action.

Enrofloxacin Sorption on Smectite Clays: Effects of PH, Cations, and Humic Acid

Journal of Colloid and Interface Science. Jan, 2012  |  Pubmed ID: 22285099

Enrofloxacin (ENR) occurs widely in natural waters because of its extensive use as a veterinary chemotherapeutic agent. To improve our understanding of the interaction of this emerging contaminant with soils and sediments, sorption of ENR on homoionic smectites and kaolinite was studied as a function of pH, ionic strength, exchangeable cations, and humic acid concentration. Batch experiments and in situ ATR-FTIR analysis suggested multiple sorption mechanisms. Cation exchange was a major contributor to the sorption of cationic ENR species on smectite. The decreased ENR sorption with increasing ionic strength indicated the formation of outer-sphere complexes. Exchangeable cations significantly influenced the sorption capacity, and the observed order was Cs

Correlation of IDH1 Mutation with Clinicopathologic Factors and Prognosis in Primary Glioblastoma: A Report of 118 Patients from China

PloS One. 2012  |  Pubmed ID: 22291938

It has been reported that IDH1 (IDH1R132) mutation was a frequent genomic alteration in grade II and grade III glial tumors but rare in primary glioblastoma (pGBM). To elucidate the frequency of IDH1 mutation and its clinical significance in Chinese patients with pGBM, one hundred eighteen pGBMs were assessed by pyro-sequencing for IDH1 mutation status, and the results were correlated with clinical characteristics and molecular pathological factors. IDH1 mutations were detected in 19/118 pGBM cases (16.1%). Younger age, methylated MGMT promoter, high expression of mutant P53 protein, low expression of Ki-67 or EGFR protein were significantly correlated with IDH1 mutation status. Most notably, we identified pGBM cases with IDH1 mutation were mainly involved in the frontal lobe when compared with those with wild-type IDH1. In addition, Kaplan-Meier survival analysis revealed a highly significant association between IDH1 mutation and a better clinical outcome (p = 0.026 for progression-free survival; p = 0.029 for overall survival). However, in our further multivariable regression analysis, the independent prognostic effect of IDH1 mutation is limited when considering age, preoperative KPS score, extent of resection, TMZ chemotherapy, and Ki-67 protein expression levels, which might narrow its prognostic power in Chinese population in the future.

A Naturally Occurring Nonapeptide Functionally Compensates the CP1 Domain of Leucyl-tRNA Synthetase to Modulate Aminoacylation Activity

The Biochemical Journal. Feb, 2012  |  Pubmed ID: 22292813

Aminoacyl-tRNA synthetases (aaRSs) establish the rules of the genetic code by catalyzing the formation of aminoacyl-tRNA. The quality control for aminoacylation reaction is achieved by editing activity, which is usually carried out by a discrete editing domain. For leucyl-tRNA synthetase (LeuRS), the connective peptide 1 (CP1) domain is the editing domain responsible for hydrolyzing mis-charged tRNA. The CP1 domain is universally present in LeuRSs except LeuRS from Mycoplasma mobile (MmLeuRS). The substitute of CP1 in MmLeuRS is a nonapeptide (MmLinker). We show here that the MmLinker, which is critical for aminoacylation activity of MmLeuRS, could confer remarkable tRNA charging activity to the inactive CP1-deleted LeuRS from Escherichia coli (EcLeuRS) and Aquifex aeolicus (AaLeuRS). Furthermore, CP1 from EcLeuRS could functionally compensate the MmLinker and endow MmLeuRS with post-transfer editing capability. These investigations provide a mechanistic framework for the modular construction of aaRSs and their coordination to achieve catalytic efficiency and fidelity. These results also show that the pre-transfer editing function of LeuRS originates from its conserved synthetic domain, and shed light on future mechanism study.

An Analysis of Nonlinear Vibrations of Coupled Thickness-shear and Flexural Modes of Quartz Crystal Plates with the Homotopy Analysis Method

IEEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control. Jan, 2012  |  Pubmed ID: 22293733

We investigated the nonlinear vibrations of the coupled thickness-shear and flexural modes of quartz crystal plates with the nonlinear Mindlin plate equations, taking into consideration the kinematic and material nonlinearities. The nonlinear Mindlin plate equations for strongly coupled thickness- shear and flexural modes have been established by following Mindlin with the nonlinear constitutive relations and approximation procedures. Based on the long thickness-shear wave approximation and aided by corresponding linear solutions, the nonlinear equation of thickness-shear vibrations of quartz crystal plate has been solved by the combination of the Galerkin and homotopy analysis methods. The amplitudefrequency relation we obtained showed that the nonlinear frequency of thickness-shear vibrations depends on the vibration amplitude, thickness, and length of plate, which is significantly different from the linear case. Numerical results from this study also indicated that neither kinematic nor material nonlinearities are the main factors in frequency shifts and performance fluctuation of the quartz crystal resonators we have observed. These efforts will result in applicable solution techniques for further studies of nonlinear effects of quartz plates under bias fields for the precise analysis and design of quartz crystal resonators.

Glioblastoma with an Oligodendroglioma Component: Distinct Clinical Behavior, Genetic Alterations, and Outcome

Neuro-oncology. Feb, 2012  |  Pubmed ID: 22326863

Glioblastomas (GBMs) containing foci that resemble oligodendroglioma are defined as GBM with oligodendroglioma component (GBMO). However, whether GBMO is a distinct clinicopathological variant of GBM or merely represents a divergent pattern of differentiation remains controversial. We investigated 219 consecutive primary GBMs, of which 40 (18.3%) were confirmed as GBMOs. The clinical features and genetic profiles of the GBMOs were analyzed and compared with the conventional GBMs. The GBMO group showed more frequent tumor-related seizures (P= .027), higher frequency of IDH1 mutation (31% vs. <5%, P= .015), lower MGMT expression (P= .016), and longer survival (19.0 vs. 13.2 months; P= .022). In multivariate Cox regression analyses, presence of an oligodendroglioma component was predictive of longer survival (P= .001), but the extent of the oligodendroglial component appeared not to be linked to prognosis (P= .664). The codeletions of 1p/19q, somewhat surprisingly, were infrequent (<5%) in both GBMO and conventional GBM. In addition, the response to aggressive therapy differed: the GBMO group had no survival advantage associated with aggressive treatment protocols, whereas a clear treatment effect was observed in the conventional GBM group. Collectively, the clinical behavior and genetic alterations of GBMO thus differs from those of conventional GBM. Presence of an oligodendroglial component may therefore be a useful classification and stratification variable in therapeutic trials of GBMs.

Seizure Characteristics and Outcomes in 508 Chinese Adult Patients Undergoing Primary Resection of Low-grade Gliomas: a Clinicopathological Study

Neuro-oncology. Feb, 2012  |  Pubmed ID: 22187341

Seizure is a common presenting manifestation and plays an important role in the clinical presentation and quality of life for patients with low-grade gliomas (LGGs). The authors set out to identify factors that influence preoperative seizure characteristics and postoperative seizure control. Cases involving adult patients who had undergone initial surgery for LGGs in a single institution between 2005 and 2009 were retrospectively reviewed. Univariate and multivariate logistic regression analyses were used to identify factors associated with preoperative seizures and postoperative seizure control. Of the 508 patients in the series, 350 (68.9%) presented with seizures. Age less than 38 years and cortical involvement of tumor were more likely to be associated with seizures (P = .003 and .001, respectively, multivariate logistic analysis). For the cohort of 350 patients with seizures, Engel classification was used to evaluate 6- and 12-month outcome after surgery: completely seizure free (Engel class I), 65.3% and 62.5%; not seizure free (Engel classes II, III, IV), 34.7% and 37.5%. After multivariate logistic analysis, favorable seizure prognosis was more common in patients with secondary generalized seizure (P = .006) and with calcification on MRI (.031). With respect to treatment-related variables, patients achieved much better seizure control after gross total resection than after subtotal resection (P < .0001). Ki67 was an independent molecular marker predicting poor seizure control in the patients with a history of seizure if overexpressed but was not a predictor for those without preoperative seizures. These factors may provide insight into developing effective treatment strategies aimed at prolonging patients' survival.