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Coxsackievirus B3-induced apoptosis and Caspase-3

Cell Research 2003年第3期13卷 203-209页

作者： jian ping yuan,wei zhao,hong tao wang,kai yu wu,tao li,xiao kui guo,shan qing tongdepartment of microbiology and parasitology,shanghai second medical university,shanghai 200025,china Department of microbiology and parasitology Shanghai Second Medical University Shanghai China

Cell death can be classified into two categories: apoptosis and necrosis. Apoptotic pathway can beeither caspase-dependent or caspase-independent. Caspase-independent cytopathic effect (CPE) has beendescribed. In order to evaluate the pattern of HeLa cell death induced by Coxsackievirus B3 (CVB3)and whether apoptosis involves caspase activation, we co-cultivated HeLa cells with CVB3 and detectedthe cytopathic changes, the alteration of mRNA and protein expression of caspase-3 gene plus caspase-3activity, as well as analyzing DNA fragmentation before and after caspase-3 activity inhibition. Accordingto the results, we propose that CVB3 may induce apoptosis and necrosis in HeLa cells, the latter appearingmuch earlier. Caspase-3 is activated at the levels of both transcription and translation, and procaspase-3 isproteolytically cleaved, thus leading to the continuous increasing of both caspase-3 precursor protein and itssubunit. However, besides CPE, apoptosis induced by CVB3 is not a direct consequence of

关键词： Coxsackievirus B3 (CVB3) apoptosis. caspase-3.

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Deletion of Helicobacter pylori vacuolating cytotoxin gene by introduction of directed mutagenesis

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World Journal of Gastroenterology 2003年第10期9卷 2251-2257页

作者： jian-ping yuan,tao li,xiao-Dong Shi,Bao-yu Hu,Gui-Zhen Yang,shan-qing Tong,xiao-kui guo,Department of microbiology and parasitology,shanghai second medical university,shanghai 200025,china. Department of microbiology and parasitology Shanghai Second Medical University Shanghai 200025 China.

AIM: To construct a vacA-knockout Helicobacter pylori mutant strain, whose only difference from the wild strain is its disrupted vacA *** AND RESULTS: A clone containing kanamycin resistance gene used for homologous recombination was constructed in a directional cloning procedure into pBluescript ⅡSK, and then transformed into vacA+ H pylori by *** growing on the selective media containing kanamycin were harvested for chromosomal DNA extraction,and the allelic exchange was determined by polymerase chain reactions and sequencing. Loss of vacuolating activity of the vaci-knockout strain was confirmed by examining the gastric cells co-cultured with cell-free supernatants from H pylori wild strain or the ***: We constructed a vacA-knockout strain of H pylori through direct mutagenesis, which creates an important precondition for the future research on virulence comparison with gene expression analysis.

关键词： HP感染基因缺失基因突变细胞毒素克隆

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