Role of phosphoinositide pathway in the formation of cardiac hypertrophy induced by pressure overload in rats / 中国病理生理杂志

AIM: To investigate the role of phosphoinositide pathway in the formation of pressure-overload cardiac hypertrophy. METHODS: Cardiac hypertrophy was induced in male Sprague-Dawley rats with coarctation of abdominal aorta, whole heart weight/body weight ratio was tested after 10 or 30 days of operation. Content of G?q/11 protein in left ventricle was detected by immunoblot analysis and concentration of IP 3 was measured by radioimmunoassay. RESULTS: At 10 and 30 days, whole heart weight/body weight ratio of coarctation aorta (CA) group was higher than that of sham-operated (SO) rats ( P 0.05). At 10 days, the level of IP 3 significantly increased in left ventricle of CA rats compared with the control animals ( P

Cloning of human sperm protein(SP17) and expression in escherichia coli DH5? / 中国病理生理杂志

AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmid for expression in E. coli. METHODS: Total fragment of hSP17 cDNA gene were amplified by RT-PCR, then subcoloned into pGEX-3b to generate recombinant hSP17/pGEX. Right orientation of insert are identified by restricted enzyme digestion. Transform the correct recombinant plasmid into the E. coli DH5a. The expression of fusion proteins hSP17-GST were induced by adding isopropylthiogalactoside (IPTG). RESULTS and CONCLUSION: The recombinant plasmid hSP17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected.

Interleukin-6 induced upregulation of basic fibroblast growth factor isoform through phorbol ester-sensitive protein kinase C in rat vascular smooth muscle cells / 中国病理生理杂志

AIM: The role of protein kinase C(PKC) in the effect of Interleukin-6(IL-6) on basic fibroblast growth factor(bFGF) expression was investigated in rat vascular smooth muscle cells(VSMC). METHODS: Western-blotting was adopted to observe the variation of bFGF and its receptor type I isoforms expression. RESULTS: IL-6 increased all the three basic fibroblast growth factor isoforms in a dose-dependent (0-10.0 ?g/L) manner. The upregulatory activities peaked at 24 h as demonstrated . In addition, after exhaustion of intracellular phorbol ester-sensitive PKC, the upregulatory effects of IL-6 on bFGF exprssion in VSMC declined ( P

SURFACE EXPRESSION AND COLOCALIZATION OF NMDA RECEPTOR AND AMPA RECEPTOR ON DENDRITE OF HIPPOCAMPAL NEURONS IN CULTURE / 解剖学报

Objective To investigate the developmental profiles on surface expression and colocalization of NMDA receptor and AMPA receptor on dendritic structures in cultured hippocampal neurons of rats. Methods Two vectors expressing green fluorescent protein tagged GluR1 subunit(GFP-GluR1) and FLAG tagged NR2B subunit(FLAG-NR2B) were co-transfected into cultured hippocampal neurons at 5 days in vitro(DIV5).Surface expressed GFP-GluR1 containing AMPA receptor clusters and FLAG-NR2B containing NMDA receptor clusters were then labeled in living neurons by using anti-GFP primary antibody followed by Alex488-conjugated secondary antibody,and anti-FLAG primary antibody followed by Cy3-conjugated secondary antibody.Furthermore,distribution of the surface clusters was observed on the detailed dendritic structures visualized with co-expression of cyan fluorescent protein tagged actin(CFP-Actin). Results The numbers for GluR1 containing AMPA receptor clusters,NR2B containing NMDA receptor clusters,and colocalized receptor clusters per 100*!?m dendrite in DIV7 and DIV14 neurons were 59.2?5.6 and 74.8?3.1(P