Aim To investigate roles of ROS in oxaliplatin-induced PUMA expression and apoptosis in colon cancer cells.Methods ROS was used as an oxidative stress in vitro and PUMA expression was determined by Western blot analysis,the apoptosis induced by ROS was assessed by Hoechst 33258 dye staining,the proliferation of the colon cancer cells treated with oxaliplatin and antioxidant was determined by MTT assay.Results ROS induced apoptosis and PUMA expression in colon cancer cells;suppression of PUMA expression by stable transfecting PUMA anti-sense vector decreased apoptosis induced by ROS;oxaliplatin-induced PUMA expression was abrogated by antioxidant and the proliferation of colon cancer cells treated with oxalipla-tin was increased by antioxidant.Conclusions Our data suggests that PUMA and ROS play important roles in oxaliplatin-induced apoptosis.Oxaliplatin induces PUMA expression and apoptosis partly through ROS in colon cancer cells.

Objective To investigate the effects of catalase on expression of cytokines and activation of nuclear factor-?B in the intestinal mucosa in rat with ulcerative colitis(UC). Methods UC was reproduced in rats with the oral innoculation of dextran sulfate sodium (DSS).The expressions of TNF-?, IL-1? and IL-8 in the intestinal mucosa of rats were respectively determined by a semi-quantatitive assay, RT-PCR. The activation of NF-?B in the intestinal mucosa were assessed with electrophoretic mobility shift assay (EMSA). Results Comparing the mice challenged with DSS alone and to those treated with catalase, both symptoms and the lesions in the colonic mucosa were milder in the animals pretreated with catalase during the induction of colitis than that of control group and catalase treatment after induction of colitis group. Furthermore, the expressions of TNF-?, IL-1?, IL-8 were significantly down-regulated(P

Objective To evaluate the expression of Fas Ligand (FasL) on gastric T cell during H.pylori infection and its cytotoxicity to gastric epithelial cells. Methods FasL protein expressions on mono nuclear cells of gastric mucosa with and without H.pylori infection were examined by immunohistochemistry assay. FasL mRNA expressions were detected in gastric T cell lines isolated from H.pylori infected and uninfected gastric biopsies by RT PCR. The function of FasL expressed by gastric T cells to induce apoptosis in Fas bearing cells was determined by a co culture bioassay (JAM), while the Fas mediated apoptosis of gastric epithelial cells induced by gastric T cells were also evaluated by the same assay. Results FasL was expressed by the mononuclear cells accumulated in gastric mucosa with H.pylori infection, whereas the few mononuclear cells presented in uninfected gastric tissues were negative for FasL. FasL mRNA was detected in gastric T cells isolated from H.pylori infected gastric biopises, while that of uninfected gastric T cell lines was either negative or weak positive. Gastric T cells were capable of inducing apoptosis of Fas positive jurkat cells, while this could be blocked by Fas blocking antibody, ZB4. Conclusions T cells presented in gastric mucosa during H.pylori infection could damage gastric epithelium through Fas/FasL interaction.

Objective To construct a highly expresses catalase of Helicobacter pylori (H.pylori) and to test it's activity. Methods The catalase DNA was amplified from H.pylori chromosomal DNA by PCR techniques, inserted into the prokaryotie expression vector pET 22b(+) and then transformed into the BL21(DE3) E.coli strain which expressed catalase recombinant protein. Then the activity of H.pylori catalase was assayed by the Beers and Sizers. Results DNA sequence analysis showed that the sequence of catalase DNA was the same as that published by GenBank. The catalase recombinant protein amounted to 24.4% of the total bacterial protein after inducing with IPTG for 3 hours at 37?C and the activity of H.pylori catalase was high in the BL21 (DE3) E.coli strain. Conclusions A clone of expressing high activity H.pylori catalase has been obtained, and thus a good foundation for further study has been laid.

Objective To study the changes of serum soluble tumor necrosis factor receptor Ⅰ (sTNFR Ⅰ) and immunosuppressive acidic protein (IAP) of patients with pancreatic cancer before and after intra arterial chemotherapy and evaluate their significance. Methods The levels of sTNFR Ⅰ and IAP of 55 cases with pancreatic cancer before and after intra arterial chemotherapy were measured by enzyme linked immunosorbent assay (ELISA) and one direction immunodiffusion test respectively and compared them with those of healthy controls. Results The levels of sTNFR Ⅰ and IAP of patients with pancreatic cancer before intra arterial chemotherapy were higher than those of healthy controls ( P

Objective To construct a recombinant strain which expresses adhesin AlpA of Helicobacter pylori(H.pylori) and to study the immunogenicity of adhesin AlpA. Methods The adhesin AlpA DNA was amplified by PCR and inserted into the prokaryotie expression vector pET-22b(+)and expressed in the BL21(DE3) E.coli strain. Adhesin AlpA immunogenicity was studied by Western blot test. Results DNA sequence analysis showed that the sequence of adhesin AlpA DNA was almost the same as that of the published by GenBank. The adhesin AlpA recombinant protein accounted for 31.9% of the total bacterial protein. Western blot analysis of rAlpA confirmed that it could be specially recognized by serum from rabbit immunized with AlpA itself and H.pylori infected patients. Conclusion Cloning and high-expression of adhesin AlpA and primary confirmation of its immunogenicity lay the foundation for H.pylori gene engineering vaccine preparation and adhering mechanism research.

To study the changes in serum immunosuppressive acidic protein (IAP) and T lymphocytes rDNA transcription activity in peripheral blood of patients with pancreatic cancer before and after operation. The level of IAP and T lymphocytes rDNA transcription activity in peripheral blood of 58 patients with pancreatic cancer were measured before and after opreration by one directional immunodiffusion test and a cell image analysis of Ag NORs, respectively, and the results were compared with that of healthy controls. The level of IAP was higher while T lymphocytes rDNA transcription activity was lower before operation compared with healthy controls( P 0 05). The levels of IAP and T lymphocytes rDNA transcription activity were closely related to tumor invasion, metastasis and clinical stages (TNM). Serial determination of the levels of IAP and T lymphocytes rDNA transcription activity might serve as an important index for evalution of tumor invasion and metatasis, and also prognosis for the patients with pancreatic cancer

To study adhesion of human bifidobacterial strains to cultured human intestinal epithelial cells in vitro mediated by purified adhesin. Results showed that the binding of human bifidobacterial strains to human intestinal epithelial cells appeared to be specific. The adhesion was time and dose-dependent.These data suggested adhesion of human bifidobacterial strains to cultured human intestinal epithelial cells in vitro can be mediated specifically by purified adhesin.

To find novel antihepatitis drugs, a series of nitrate-oleanolic acid (OA) hybrids (10a, 10b, 11a-11e and 12a-12c) were designed and synthesized on the basis of previous studies using OA as lead compound, which is widely found in natural plants and liver-specific metabolism. In the present study, ten novel NO-releasing derivatives of OA were synthesized by connecting nitrate to the OA-3-OH through varying lengths of linkers containing antioxidants which were designed to increase the ability of these target compounds to scavenge free radicals. The structures of these objective compounds were determined by IR, MS, 1H NMR and elemental analysis. Their protective effects on anti-Fas mediated HepG2 cell apoptosis were in vitro evaluated by LDH assay. Compound 12a is the most potent inhibitor. Its effect on anti-Fas mediated HepG2 cell apoptosis and amount of NO-releasing in vitro are similar to those of positive control NCX-1000.

Background and purpose:The Smac/DIABLO(the second mitochondrial derived activator of caspase/direct IAP binding protein with low pI)is a new kind of mitochondria protein,it inhibits IAPs(inhibitors of apoptosis protein),including Survivin,and activates caspase-9 and caspase-3,accordingly promotes apoptosis.In this study,we investigated the effect of over-expressing mitochondrial protein-Smac/DIABLO while silencing inhibitor of apoptosis protein-Survivin on the cell growth,cell cycle and apoptosis of human colon cancer cells through cotransfecting both genes.Methods:Constructed survivin-shRNA-EGFP plasmid was transected with Smac-pcDNA3.1 plasmid into Lovo cells at either half each dose or total dose,respectively.Western blot was used to survey the protein level of Smac and Survivin;Hoechst 33258 staining was used to detect the karyomorphological diversity of apoptotic cells and evaluate apoptotic ratio roughly;PI staining and flow cytometry analysis were used to examine cell cycle;caspase-3 Detection Kits was used to detect the activity of caspase-3.Results:The expression of Smac protein increased but Survivin protein decreased 48 hr after transfection.Karyomorphological diversify of apoptotic cells were obviously observed by hoechst 33258 staining.The apoptosis rate in Smav+Survivin shRNA group was(18.5?1.7)%,which was higher than that in Smac group(9.6?1.8)% and Survivin shRNA group(15.0?0.3)%,and all of three groups were significantly higher than the control groups;The cells of G0/G1 phase increased to(51.0?6.2)% in Smac group,and the cells of S stage increased to(53.3?1.3)% in Survivin shRNA group(P