Effect of SJAMP on apoptosis of human hepatocellular carcinoma cell line HepG2 and the expression of Bcl-2, nm23-H1 in vitro / 国际外科学杂志

Objective Through studying the apoptosis induced by stichopus japonicus acid mucopoly saccharide in the hepatocellular carcinoma cell line HepG2 in vitro, analysing the expression of Bcl-2 and nm-23in HepG2, to provide the theory foundation and its feasibility on whether it can be used for the chemotherapy of hepetocellular carcinoma. Methods The cells of HepG2 were cultured in vitro and treated with SJAMP at different doses(0.25,0. 5,1.0,2.0,4.0 g/L). MTT was used to observe the inhibitory effects of SJAMP on cell growth, Western blotting was used to detect apoptosis, and the apoptosis related change of expression of protein Bcl-2 and nm23-H1. Results (1) MTT identified that SJAMP produced an obvious time-and-dose-dependent inhibitory effect on the HlepG2 cells. (2) Western blot showed that SJAMP could induce the apoptosis of HepG2 cells through changing the expression of the protein of Bcl-2 and nn23-H1 (P<0.05). Conclusion (1)SJAMP produced obvious inhibitory effects on HepG2 cells and induce HepG2 apoptosis. (2)SJAMP can enduce the anti-tumor function in the method of changing the expression of protein Bcl-2 and nm23-H1.

Co-culture of mouse blastocysts and their epigenetic modification / 生物工程学报

To discuss the effect of co-culture on the quality of mouse blastocysts and their epigenetic modification. We divided mouse zygotes into three co-culture experiment groups : with granular cells (group I ), oviduct epithelium cells (group II) and oviduct tissue (group III). Meanwhile, we set up control A (cultured in vitro, only KSOM (KCl+ simplex optimized medium)) and control B (cultured in vivo). Then we compared cleavage rate and blastocyst rate among different groups. After that we evaluated the quality of blastocysts by using ICM/TE (Inner cell mass/Trophectoderm cells) ratio via staining with propidium iodide and Hoechest333258, and analyzed the level of genome methylation and histone acetylation by immunofluorescence. Compared with the control group A, the co-culture groups had increased cleavage rate and blastocyst rate (P < 0.05), blastocyst cells and the ICM/TE ratio of co-culture groups were higher (P < 0.05), the level of genome methylation and histone acetylation had no significant difference between groups in vitro (P > 0.05), but the level of genome methylation in vivo was significantly higher than that of in vitro (P < 0.05). The co-culture methods can successfully promote the development rate of embryos in vitro, and improve the quality of the blastocyst. However, the methods have drawbacks in changing the abnormal genome methylation with in vitro culture.

Construction of HBD-3 gene mammary-specific expression vector and eukaryotic expression / 生物工程学报

To establish human beta-defensin-3 gene transgenic cell lines as competent donor cells for the production of transgenic animals using somatic cell nuclear transfer (SCNT). Firstly, we obtained human beta-defensin-3 by RT-PCR from human placenta, and subsequently inserted the fragment hBD into the corresponding site of the plasmid pBCP. Then we moved the combined fragment BCD (including 5' and 3' regulating region of beta-casein and hBD) into the corresponding site of the plasmid pEGFP-C1. Finally we successfully constructed mammary-specific expression vector pEBCD. We transected pEBCD into Holstein Fetal fibroblast cells by Lipofectamine TM-2000 and selected in medium with G418 for three to four weeks. We identified G418 resistant transfectants by PCR, RT-PCR and EGFP detection. Our results indicated that human beta-defensin-3 gene stably was integrated into the open region of the chromatin in G418 resistant fibroblast cells. Meanwhile we identified the expression of human beta-defensin-3 in the supernatant of stable transfected mammary epithelial cells by Western blotting. This study may provide competent transgenic donor cells for the production of transgenic animals by SCNT and improve the efficiency of transgenic cloning.

Cause of death after TACE in China during the past 14 years / 国际外科学杂志

Objective To study the cause of death and mechanism after(TACE)in China during the past 14 years.Methods Related repots in Chinese Medical Current Content(CBM)and National Knowledge lnfrastruc ture(CNKI)from January 1994 to June 2008 were retrieved.The cause of death and mechainsm after TACE wer e analyzed.Results A total of 150 patients who died after TACE were reposed in China during the past 14 ye ar s.84%eases were caused by liver lunction failure,upper gastrointestinal bleeding and rupture of liver cancer. 78.7%cases died one month postoperation.Conclusion Liver function failure.upper gastrointestinal bleeding and rupture of liver cancer are the main complications which Can cause death and the majority cases died early.

The effect of KLT on apoptosis of HepG2 cells and expression of Bcl-2 and Caspase-8 / 国际外科学杂志

Objective To investigate the effects of KLT on apoptosis of HepG2 cells and expression of Bcl-2 and Capase-8. Methods The cell fine HepG2 was induced by diverse density of KLT, and HepG2 cell was collected respectively after induction of 12 h, 24 h, 48 h. The control group was installed simulta-neon]y. The cell apoptosis and the expression of Bcl-2 and Caspase-8 were detected by flow cytometry (FCM). Results KLT can induce the apoptosis of HepG2 cell significantly, and the longer time past, the more apoptosis of HepG2 was. KLT can increase the expression of Caspase-8, but ineffective to Bcl-2. Con-clusion KLT can significantly induce the apoptosis of HepG2 cell through regulating the expression of Caspase-8.