Background and purpose: Bladder cancer is the most common urological malignancy in our country which seriously threatens human health, and its incidence increased year by year. This study aimed to investigate the effects of hinokitiol on the proliferation, apoptosis and autophagy in human bladder cancer cell lines. Methods:CCK-8 assays were performed to analyze the effects of hinokitiol on the proliferation of J82 cells. Apoptosis rate was determined by lfow cytometry. Cleaved caspase 3, LC3 and P62 protein expression was determined using Western blot. EGFP-LC3 microscopy assay was performed to assess autophagy. Results: Hinokitiol significantly inhibited the proliferation of J82 cells and induced cell apoptosis via caspase pathway. The apoptosis effect of hinokitiol could be partially antagonized by Z-VAD-FMK. Hinokitiol induced autophagy of J82 cells, increased LC3 expression and down-regulated P62 expression. 3-MA is able to rescue Tet-induced cell death. Conclusion:Hinokitiol can inhibit the proliferation of J82 cells and induce cell apoptosis via autophagy activation.

Objective To clone cDNA of trichosanthin (TCS) and purify TCS, and to study its influence on apoptosis and growth inhibition of colorectal carcinoma LoVo cells in vitro. Methods MTT assay was adopted to measure the growth inhibition ratio of LoVo cells treated with TCS, and apoptosis was assayed by agarose gel eletrophoresis. Results The results showed that the higher concentration of TCS and the longer testing time, the stronger growth inhibition of LoVo cells. DNA agarose gel eletrophoresis showed a gradient, which confirmed that TCS could induce the apoptosis of LoVo cells. Conclusions TCS can inhibit the growth of LoVo cells in vitro and induce its apoptosis. Our study provides evidence for the application of TCS in clinical treatment of human colorectal carcinoma.

Objective:To investigate the effect of calcium ionorphore on B7 costimulatory molecules of chronic myeloid leukemia cells line K562.Methods:Well growing K562 cells were cultured in the medium containing calcium ionorphore(375 ng/ml),with K562 cells without CI treatment as control.The cells′ viability and number were calculated by Trypan Blue exclusion at 0,48 and 96 h.Before and after 96 h of cultured,B7-1 and B7-2 expression was assayed by flow cytometry.The proliferation of allogeneic human T cells was measured by MTT colorimetry.Results:B7 costimulatory molecules were abcent or lowly expressed on K562 cells.After 96 h of CI treatment,B7 costimulatory molecules of K562 cells were markedly upregulated and marked activation of allogeneic T cells occurred.No notable morphological change was found during the culture.K562 cells cultured in medium with CI grow slowly than that without CI.Conclusion:B7 costimulatory molecules expression on chronic myeloid leukemia cells line K562 surface was defective.These costimulatory molecules on K562 cells can be upregulated by calcium ionorphore.Calcium ionorphore may inhibit the growth of K562 cells.

To obtain the functional fusion protein of rhoptry protein 2, compound rhoptry protein2 and surface antigen 1 of Toxoplasma gondii. the ROP2 and P30 genes from genomic DNA of T.gondii RH strain were amplified by PCR, and were inserted into pMD18-T cloning vector. Then the ROP2 fragment was subcloned to pET-30a(+) plasmid digested by EcoRⅠand Hind Ⅲ to construct plasmid pET-ROP2. Furthermore,the P30 fragment was subcloned into pET-ROP2 digested by BglⅡand EcoRⅠto create plasmid pET-ROP2-P30, the resulting recombinant plasmids , transformed into E.coli BL21 (DE3), were induced with IPTG. and the proteins identified by SDS-PAGE were further purified and refolded. The biological activity was analyzed by Western blot with specific antibody. It was found that the sizes of ROP2 and ROP2-P30 were 1212 and 1896bp with corresponding molecular weight 50- kDa and 75-kDa, respectively. The recombinant protein ROP2 (50-kDa) could specifically react with rabbit-polyclonal antiserum, and complex fusion protein ROP2-P30 (75- kDa) could react with P30 monoclonal antibody.