Culture and identification of human and rabbit corpus cavernosum smooth muscle cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate an effective method to produce large numbers of pure corporal smooth muscle cells in vitro according to the requirement of study.</p><p><b>METHODS</b>In this study, we used the primary tissue culture technique to isolate and culture the corpus cavernosum smooth muscle cells (CCSM) from human males with normal erectile function and New Zealand white rabbits. The cells were identified in regard to morphological and growing characteristics via immunohistochemical methods (including alpha-smooth muscle actin, desmin, myocin and factor VIII related antigen), special dye techniques (including Masson and Van Gieson) and transmission electron microscope.</p><p><b>RESULTS</b>CCSM were isolated and cultured successfully with high purity. Morphologically, the cells were spindle shaped and grow on top of each other, resembling a "hill and valley" in appearance. When characterized in immunohistochemistry, the cells were stained with alpha-smooth muscle actin, desmin and myocin, but not with anti-factor VIII, an endothelial marker.</p><p><b>CONCLUSION</b>The CCSM, which can be isolated and cultured successfully, may be used for further studying their biological function. The CCSM cultured in vitro was proved to be useful to evaluate and investigate the effect of some new medicine for penile erection. There is also a clinical and theoretical significance in further studying the experimental mechanisms of erectile dysfunction.</p>

The infectivity of the recombinant vaccinia virus expressing human interleukin-2 and its effect on MKN45 gastric cancer cells in vitro / 中华消化杂志

Objective To study the effect of the recombinant vaccinia virus expressing human interleukin 2 on MKN45 gastric cancer cells in vitro. Methods Recombinant vaccinia virus expressing hIL 2 (VMJ601hIL 2) was constructed by homologous recombination using molecular virology, and VMJ601hIL 2 was detected by DNA hybridization technique and the recombinant gene product was analyzed by SDS PAGE. In addition, MKN45 gastric cancer cells was infected by VMJ601hIL 2 and the effect of VMJ601hIL 2 on the gastric cancer cells was evaluated in vitro. Results hIL 2 protein with biological activity can be secreted by MKN45 gastric cancer cells after heavily infected by VMJ601hIL 2. Conclusions It is one of the crucial steps that VMJ601hIL 2 has been constructed and identified since it forms essential prerequisite for further in vivo gene therapy of gastric cancer.

Effects of Helicobacter pylori on expression of DNA mismatch repair genes in AGS cell line / 中华消化杂志

Objective To investigate the effects of Helicobacter priori (H.prlori) strains from the patient with gastric cancer on the expression of DNA mismatch repair (MMR) genes in AGS cells in vitro. Methods AGS cells were co-cultured with ten strains of H.prlori from five patients with gastric cancer and five patients with gastritis respectively.The expressions of DNA MMR genes (hMSH2 and hMLH1) in mRNA and protein levels were determined by RT-PCR and Western blot.The enteropathogenic E.coli served as a bacterial control.Results E.coli and H.priori strains from gastritis have no effects on the expression of hMSH2 and hMLH1 in both protein and mRNA levels on the whole,while all the H.prlori strains from gastric cancer reduced expression levels of hMSH2 and hMLH1.Conclusions There are different effects between H. prlori strains from gastric cancer and those from gastritis on DNA MMR in AGS cell line,indicating that infec- tion of some H.prlori strains might lead to the inhibition of DNA MMR,and that in turns increases the risk of gastric carcinogenesis during chronic H.prlori infection.

Expression of hepatocyte growth factor and c-met stimulated by high glucose in human kidney fibroblast and its significance / 中华内分泌代谢杂志

Objective To observe high glucose induced expression of hepatocyte growth factor (HGF) and c met in human kidney fibroblast. Methods The effects of glucose concentrations on expression of HGF, c met and plasminogen activator inhibitor (PAI) 1 in cultured human kidney fibroblasts were observed by RT PCR. In the same system, the effect of exogenous HGF on the expression of PAI 1 was investigated. Results Human kidney fibroblasts cultured in high glucose concentration (25 mmol/L) showed higher HGF and c met expressions in the early stage and then manifested a gradient decrease of HGF and c met expressions, but PAI 1 expression was gradiently increased. Exogenous HGF resulted in inhibiting PAI 1 expression. Conclusion HGF is a potential anti fibrogenic factor and activates matrix degradation pathways in diabetic kidney by reducing PAI 1 expression.