Publications (38) View all
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Article: Effect of neuronal excitotoxicity on Munc18-1 distribution in nuclei of rat hippocampal neuron and primary cultured neuron.
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ABSTRACT: Munc18-1 has an important role in neurotransmitter release, and controls every step in the exocytotic pathway in the central nervous system. In the present study, whether epileptic seizure causes a change of Munc18 localization in neuronal nuclei was analyzed. Epilepsy models were established by injection of kainic acid (KA) solution into hippocampus of Sprague-Dawley (SD) rats or intraperitoneal injection of KA in Kunming mice. The hippocampal neurons were prepared from embryonic day 18 SD rats, and cultured in neurobasal medium, followed by treatment with glutamate for 3 h. Neuronal and glial nuclei of hippocampus were separated by sucrose density gradient centrifugation. The nucleus-enriched fractions were stained with 0.1% Cresyl Violet for morphological assay. Immunochemistry and immunoelectron microscopy with anti-Munc18-1 antibody were used to determine the nuclear localization of Munc18-1. Immunoblotting was used to detect the protein level of Munc18-1. The localization of Munc18-1 in nucleus of rat hippocampal neuron was confirmed by immunochemistry, immunoelectron microscopy, and immunoblotting detection of neuronal nucleus fraction. In animals receiving intrahippocampal or intraperitoneal injection of KA, immunostaining revealed that the expression of Munc18-1 decreased in pyramidal cell layer of CA regions, as well as in hilus and granular cell layer of dentate gyrus in hippocampus. Moreover, immunoblotting analysis showed that the expression level of Munc18-1 in nucleus fraction of hippocampus significantly decreased in KA-treated animals. The relationship between the change of Munc18-1 expression in neuronal nuclei and neuronal over-activation was also tested in primary cultured neurons. After treatment with 50 μmol/L glutamate acid for 3 h, Munc18-1 level was decreased in nucleus fraction and increased in cytoplasmic fraction of primary cultured neurons. These results suggest that excitatory stimulation can induce the distribution change of Munc18-1 in neuron, which may subsequently modulate neuronal functions in brain.Neuroscience Bulletin 06/2011; 27(3):163-72. · 1.31 Impact Factor -
Article: Biodiesel production from rubber seed oil using poly (sodium acrylate) supporting NaOH as a water-resistant catalyst.
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ABSTRACT: Poly (sodium acrylate) supporting NaOH (NaOH/NaPAA) was prepared by in situ polymerization of aqueous solution of acrylic acid with an over-neutralization by adding excess of NaOH. NaOH/NaPAA presented a promising selectivity for water absorbency and good water retention with negligible swelling capacity in the organic solvents of methanol, glycerol, rubber seed oil methyl esters, and rubber seed oil. NaOH/NaPAA catalysts showed a basic strength of 15.0<H_<18.4 and their basicity increased with the increase of the NaOH loading amount. NaOH/NaPAA catalysts exhibited almost the same catalytic activity in the transesterification of rubber seed oil with methanol under the optimized reaction conditions compared to conventional homogeneous NaOH catalyst. Furthermore, the functional absorbent/catalyst system presented a good water resistance in the transesterification which retained high catalytic activity when a water concentration in the reaction system was less than 2 wt.%.Bioresource technology 02/2011; 102(3):2665-71. · 4.25 Impact Factor -
Article: Functional characterization of a plasma membrane Na+/H+ antiporter from alkali grass (Puccinellia tenuiflora).
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ABSTRACT: We have cloned a Na(+)/H(+) antiporter gene (GenBank accession no EF440291, PtNHA1) from Puccinellia tenuiflora (so-called alkali grass in Chinese) roots under NaCl salt stress. Its cDNA is 3775 bp and contains a 3414 bp open reading frame. The amino acid sequences of PtNHA1 show high identities with a putative plasma membrane Na(+)/H(+) antiporter from wheat. PtNHA1 was predicted to contain 11 hypothetical transmembrane domains in the N-terminal part and to localize in the plasma membrane. Genomic DNA gel blot analysis shows that PtNHA1 is a single-copy gene in the alkali grass genome. PtNHA1 is highly expressed in leaves, roots and shoots by RNA gel blot analysis. Furthermore, PtNHA1 gene expression of alkali grass was clearly up-regulated by NaCl salt stress. Overexpression of PtNHA1 in Arabidopsis resulted in enhanced tolerance of transgenic plants to NaCl stress. The ion contents analysis shows that, compared with the wild-type (WT), less Na(+) and more K(+) were accumulated in transgenic plants under NaCl stress. The results indicate that PtNHA1 play an important role in NaCl salt stress. Additionally, compared with the WT, total activities of ascorbate peroxidase (APX) and catalase (CAT), two key reactive oxygen species (ROS) detoxifying enzymes were high in transgenic plants under salt stress, respectively. The transcript levels of two APX genes (Apx1, s/mApx) and two CAT genes (Cat1, Cat2) in transgenic plants were higher than those in WT. This suggests that overexpression of PtNHA1 results in enhanced ROS-scavenging enzymes of transgenic plants under NaCl salt stress.Molecular Biology Reports 12/2010; 38(7):4813-22. · 2.93 Impact Factor -
Article: Selective blockade of cytoskeletal actin remodeling reduces experimental choroidal neovascularization.
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ABSTRACT: The efficacy of the peptide Ac-EEED on reducing cell adhesion and proliferation in vitro and choroidal neovascularization (CNV) in vivo was examined. The peptide chimera containing the Ac-EEED sequence was chemically linked to the N terminus of the XMTM delivery peptide from the E(rns) viral surface protein. Ac-EEED or scrambled control peptide (SCRAM) was added to cultures of vascular smooth muscle cells, pericytes, endothelial cells, and fibroblasts, and adhesion, growth, and matrix production was assessed. Ac-EEED or SCRAM was injected into the vitreous of mice undergoing laser rupture of Bruch's membrane to induce CNV and lesion volume, neovascularization and lesion fibrosis were assessed. Ac-EEED-induced changes in the morphology of the actin cytoskeleton by inhibiting polymerization of G-actin and disrupting the formation of stress fibers. Pretreatment with Ac-EEED resulted in endothelial cells becoming less responsive to the mitogenic and pro-adhesive effects of VEGF. Ac-EEED treatment in fibroblasts reduced TGF-β-induced fibrosis as assessed by decreased levels of connective tissue growth factor, cysteine-rich 61, collagen I (COL1A2), and collagen III (COL3A1). CNV lesion size and fibrosis were reduced in a concentration-dependent manner by up to 60%. In vitro studies showed that Ac-EEED affects a broad range of mechanical properties associated with cytoskeletal actin to reduce growth factor effects. The utilization of Ac-EEED in vivo may offer a novel therapeutic strategy by both suppressed neovessel growth and curtailing fibrosis typically associated with the involutional stage of CNV.Investigative ophthalmology & visual science 12/2010; 52(5):2490-6. · 3.43 Impact Factor -
Article: Glutamate enhances the surface distribution and release of Munc18 in cerebral cortical neurons.
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ABSTRACT: Munc18 is considered as an intracellular protein that plays an important role in exocytosis of neurotransmitters. Previous studies have demonstrated the presence of autoantibodies against Munc18 in a subgroup of Rasmussen's encephalitis patients. However, the machinery of Munc18 autoimmunity is still elusive. The present study was aimed to investigate Munc18 release from primary cultured neurons, Munc18 distribution on the outer plasma membrane of neurons, and the neurotoxicity of Munc18 antibody. The cerebral cortical neurons from embryonic day 17 Sprague-Dawley rats were prepared and cultured in neurobasal medium. The proteins in culture medium were precipitated with 10 % trichloroacetic acid, and analyzed by immunoblotting. The proteins on neuronal surface were biotinylated with EZ-Link-sulfo-NHS-LC-Biotin, and collected with avidin-conjugated agarose beads followed by immunoblotting analysis. For cell surface immunofluorescent staining, the living neurons were labeled with anti-Munc18 antibody at 4 degrees C. Neuronal injury was assessed by lactate dehydrogenase(LDH) release. Munc18 was detected in culture medium by immunoblotting analysis. After treatment with 50 micromol/L glutamate for 1 h, Munc18 content in medium was increased. Meanwhile, beta-actin and syntaxin1 were not detected in culture medium, and LDH release was not significantly increased. Moreover, glutamate enhanced Munc18 distribution on outer plasma membrane. Living neuron staining also demonstrated the localization of Munc18 on neuronal surface after glutamate treatment, especially at contacting regions between neurons. Glutamate-induced increase of surface Munc18 distribution was suppressed by NMDA receptor antagonist MK801, but not by AMPA receptor antagonist NBQX. Moreover, compared with c-Fos antibody, Munc18 antibody could induce neuronal injury, when culture medium contained the components of serum. A portion of Munc18 can be released from neurons or distributed on neuronal surface, which can be enhanced by glutamate treatment via activation of NMDA receptors. Besides, Munc18 antibody-induced neuronal injury depends on the serum components.Neuroscience Bulletin 08/2010; 26(4):273-81. · 1.31 Impact Factor
