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Objective To investigate the biological effects of Wnt gene in kidney cancer Caki?2 cells. Methods The Wnt gene was silenced in kidney cancer Caki?2 cells by lentivirus vector. The cell proliferate ability of cells in each group were assayed by CCK?8 kit at different time points. The apoptosis of Caki?2 cells was observed after silencing Wnt gene by transmission electron microscope. The invasion ability of each group cells was tested using Transwell chambers. The genes expression changes of Wnt/β?catenin signaling pathway and apoptosis related gene were determined by realtime PCR. Results Compared with the other two groups,the cell proliferate ability of the cells after silencing Wntgene was suppressed,and the difference was statistically significant(P<0.05). Apoptosis increased significantly in shRNA+Caki?2+Wnt group cells with silencing of Wntgene, and apoptotic body appeared in these cells. In invasive experiment,the number of emigrated cells in shRNA+Caki?2+Wnt group were significantly lower than other groups(P<0.05). The expression of Wnt mRNA,β?catenin mRNA and Bcl?2 mRNA in shRNA+Caki?2+Wnt group cells was lower than other groups(P<0.05). Conclusion Silencing of Wnt gene of kidney cancer Caki?2 cells can affect the proliferation rate of the cells, promote the cell apoptosis,and inhibit the invasion ability,which provide certain theoretical basis for the research and development of new drugs and new therapeutic targets.
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Objective To study to take the oxidized Mannan?modified 786?0 in renal clear cell carcinom as tumor cells antigen to sensitize Dendrit?ic cells(DC)and to observe the its killing effect on renal clear cell carcinoma of CTLs induced. Methods Getting the peripheral blood mononucle?ar cells from the volunteers,and then to be stimulated to turn to be maturation by GM?CSF and IL?4 in vitro. Taking the clear renal carcinoma cell as the tumor antigen,and then making it to be modified by oxidized Mannan to acquire the tumor cell vaccine.Experimental groups include:blank group:DC?PBS group,control group:control?DC?786?0,experimental group:DC?Ox?Mannose?786?0 group. Taking the flow cytometry to detect the changes of DC phenotype,then taking the ELISA to detect the sencretion levels of supernatant of IL?12 of DC,then taking the CCK to detecte the cytotoxicity of lymphocytes(CTL)induced by DC of these experiment groups. Results Results by flow cytometry:the mature phenotype of DCs sensitized by Ox?Mannose?786?0 group included CD80,CD83,CD86 and HLA?DR expressed significantly higher than the other groups. As well as the secretion levels of IL?12. Meanwhile the cytotoxicity activity of lymphocytes(CTLs)induced by DCs which are sensitized by Ox?Mannose?786?0 increased more significantly than the other groups. Conclusion Glycosylated Antigens can be more effective in sensitizing antigen?presenting cells DC,and stimulating them to be maturation,while the killing effect to tumor cells also have noticeably improved.
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Objective To extract exosomes from dendritic cell (DC)and investigate antitumor effect of the special cytotoxic T lymphocytes activated by exosomes against 769-P.Methods Monocytes from peripheral blood of healthy donors were cultured to induce DC in intro and their phenotypes were analyzed by FACS.The exosomes were extracted from DC by ultrafiltration and ultracentrifugation and observed under electric microscope.Proliferation of T cells and the cytotoxicities of CTL against 769-P induced by exosome were measured by MTT.Results The exosomes could be separated from culture supernatant of DC by ultrafiltration and ultracentrifugation.The combination of the exosomes from DC and the 769-P antigen could effectively stimulate T cell proliferation and enhance their cytotoxicity, the absorption value of T cell proliferation(1.68±0.22), the cytotoxicity of CTL(38.23±2.16)%.Conclusion The exosomes extracted by ultrafiltration and ultracentrifugation can present tumor antigen to T cells and induce the cytotoxicity of CTL.
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Objective To analyze the relationship between the expression of MHC class I chain-related gene A/B(MICA/B) on different human cancer cells and their sensitivity to NK cell cytotoxicity.Methods Hie expression MICA/B in K562,Bcap37,769P and A549 cells was measured by FACS.Isolation of NK cells were obtained by the modified RosetteSep~((R)) procedure.Cytotoxicity of NK cells at different ef0.50)%,(90.72±0.64)%,(55.59±0.55)%,and(3.84±0.10)% respectively.The purity of NK cells obtained by the modified RosetteSep~((R)) procedure was(96.52±2.42)%,whereas the cytotoxicity of NK cells against K562,Bcap39 and 769P was much higher than that of A549(P<0.01).The expression of MICA/B was significantly correlated with the cytotoxicity of NK cells at E:T ratios of 5:1,10:1 and 20:1 respectively(P<0.01).Conclusion The MICA/B expression on cancer cell lines was correlated with NK cell-mediated cytolysis and influenced the cytotoxicity of NK cells.
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Objective To observe whether hepatitis B vaccine enhance the treating effect of cyto-kine induced kill(CIK) cells on hepatitis B virus transgenic(HBV-Tg) mice. Methods The HBV-Tg mice were treated with CIK cells by peritoneal injection and hepatitis B vaccine by hypodermic injection. The HBV DNA level were tested by real-time PCR,T lymphocyte subgroup were detected by flow cytometry and the pathological diversify of hepatic tissue were observed by HE staining. Results The HBV DNA loading in peripheral blood of HBV-Tg mice decreased after CIK cells were treated and CD3~+ , CD4~+ and CD8~+ cells increased which were enhanced after CIK cells combined with hepatitis B vaccine. Conclusion Hepa-titis B vaccine enhanced the treating effect of CIK on HBV-Tg mice which may be implemented by increased the blood level of CD3~+, CD4~+ and CD8~+ cells, especially CD8~+ cells level.
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Objectives To evaluate the reliability and validity of SF-36 in old people in community. Methods 378 old people in community were investigated by SF-36 by face-to-face interview, and the score of items was analyzed.Reliability and validity of SF-36 were evaluated by item-internal consistency,item discriminant vadility,Cronbach's alpha coefficient, et al. Results The clustering and ordering of the item means of SF-36 were the same as that hypothesized, except for items PF2,PF10,RP3,GH4. The correlation between an item and its hypothesized scale was 0.4 or above for all items. The Cronbach's alpha coefficient were above the standard of 0.7 for all except SF scale. Conclusions SF-36 Health Survey Questionnaire is available for the elderly people. The present evaluation proposes some minor word and item changes which is more suitable for old people.