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Objective To investigate the regulation of insulin VGLUT2 gene expression in pancreatic β cells and its exactly mechanism.Methods The rat pancreatic β cell line RIN-5F was treated by different insulin concentrations and different siRNA.The changes of VGLUT2 mRNA and protein expression levels were detected by adopting real-time qPCR and Western blot.Results In-sulin with a concentration of 100,200 nmol/L significantly inhibited the expressions of VGLUT2 mRNA and protein in RIN-5RF cells(P<0.05),moreover the inhibiting effect was most significant at 100 nmol/L.After 100 nmol/L insulin treatment,the expres-sions of VGLUT2 mRNA and protein at 12,18,24 h were significantly inhibited compared with that at 0 h(P<0.05).Compared with the Blank group,Lip2000 group and Control-siRNA group,after interfering RIN-5F by using IR-siRNA,IRS1-siRNA and IRS2-siRNA,the inhibition situation of VGLUT2 mRNA and protein expressions by 100 nmol/L insulin in each group was signifi-cantly recovered(P<0.05).Conclusion Insulin at low concentration could inhibit VGLUT 2 gene expression in rat pancreatic β cell line RIN-5F.
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Objective To study the effect of 20-HETE on apoptosis in cultured neonatal rat cardiomyo-cytes and investigate its mechanism. Methods Neonatal rat cardiomyocytes were cultured in vitro.CCK-8 method was used to detect the cell activity and TUNEL assay was performed to analyze the cell apoptosis. Flou-3/AM la-belled assay was applied to measure the concentration of intracellular calcium([Ca2+]i). Western blot was per-formed to measure the expressions of RyR2,SERCA2a,CaMKII and phospho-CaMKII. Results Treatment with 20-HETE reduced the activity of cardiomyocytes and induced cell apoptosis obviously,while KN-93,an inhibitor of CaMKII,blocked the effects of 20-HETE. Treatment with 20-HETE significantly increased cardiomyocytes [Ca2+]i,up-regulated the expression of RyR2,and down-regulated the expression of SERCA2a,which could be blocked by KN-93. 20-HETE also increased the expressions of CaMKII and phospho-CaMKII in cardiomyocytes, indicating 20-HETE played a role in activating the CaMKII signaling pathway. Conclusions 20-HETE leads to altered functions of cardiac sarcoplasmic reticulum calcium-transport protein RyR2 and SERCA2a via activating the CaMKII signaling pathway,which causes calcium overload and induces apoptosis in neonatal rat cardiomyocytes.
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Objective To explore the application of scene type trace method in physiology teach-ing reform. Methods 2009 grade clinical medicine students in Zunyi Medical College were randomly di-vided into two groups:control group (n=111) and experiment group (n=108). Students in control group were taught by traditional teaching method while those in experimented group by scene type trace method. According to the questionnaire survey and the final exam statistical analysis,the effects of teach-ing were evaluated. SPSS 15.0 software was used to do statistical analysis. Qualitative data were analyzed by chi-square test of completely randomized design and two sets of frequency distribution while quantita-tive data by independent samples t test. P<0.05 means that difference is statistically significant. Results There were statistically differences in satisfaction degree of two teaching methods. More students were sat-isfied with scene type trace method than traditional teaching method. Final exam achievement in scene type trace method group(79.90±14.25,90.74%)were higher than that in the traditional teaching method group(72.35±17.09,83.78%). Conclusions Physiology teaching scene type trace method is helpful to improve the students' learning initiative and significantly improve students' learning achieve-ment.
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In theory teaching,we let students know the knowledge of some important discovery and invention in the history of physiology and while in experiment teaching,we added the experiments designed by students in addition to traditional experiments,so as to enhance the students,ability of innovation.
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Aim The aim of the present study is to in-vestigate the effect of the specific antagonist NVP-AAM077 and Ro25-6981 of NMDA receptor subunit NR2A and NR2B on global cerebral ischemia-induced neuronal injury in the CA1 region. Methods Male C57BL/6 mice subjected to global ischemia by the three-vessel occlusion( 3-VO) method,were divided into four groups: sham-operated group,ischemia/reperfusion control group,NVP-AAM077 treatment group,and Ro25-6981 treatment group. Hippocampal sections were processed for Fluoro-Jade B staining to detect degenerating neurons and for Nissl staining to identify surviving neurons. The expression of brain derived neurotrophic factor( BDNF) were measured by Western blot. Results ① Transient brain ischemia induced selective and delayed neuron death in the CA1 region of the hippocampus at 12-minute ischemia after reperfusion 3 days. NR2A subtype specific antagonist NVP-AAM077 enhanced neuronal death after transient global ischemia ( P