Objective To construct a mifepristone inducible recombinant adenovirus carrying enhanced green fluorecence protein for future gene therapy in vivo.Methods Full length red fluorescent gene and mifepristone regulator cassette was subcloned into PDC313 shuttle plasmid.The positive clone was identified by sequencing.Recombinant adenovirus were produced after cotransfection of the shutter vector PDC-EGFP and adenovirus DNA helper plasmid pBHGloxE1,3Cre into 293 cells.The recombined adenovirus was purified by CsCl density gradient centrifugation and its titer was determined by end point dilution assay.Fluorescence microscopy and FCM were confirmed by the activation of this regulatable recombinant adenovirus vector in infected cells.Results The adenovirus vector containing enhanced green fluorecence protein gene was identify by PCR.The virus titer of Ad-RUEGFP was 6.6?1010pfu/ml.Without the absence of mifepristone,no significant enhanced green fluorecence protein activation was observed,whereas in the presence of mifepristone activation of the enhanced green fluorecence protein was positive correlation with the inducer under definite range.Conclusion A replication-defective recombinant adenovirus vector containing enhanced green fluorecence protein was successfully constructed,which can be inducible by mifepristone.

Objective: To study the effect of arsenic trioxide (As2O3) on human pancreatic cancer cell proliferation and apoptosis (mainly early stage) in vitro. Methods: SW1990 cells line were trea ted with As2O3 at different concentration. Cell proliferation was evaluated by MTT and apoptosis by Annexin-Ⅴ-fluostaining, electron-microscopy, flow cytometry and immunocytochemical staining of Bcl-2 and Bax. Results: As2O3 and cisplatin had the same cytotoxity on SW1990. The cytotoxic effe ct on tumor cell was produced by induction of apoptosis. Twelve hours after cult ure with 10 μg/ml As2O3, much more SW1990 cells went into apoptosis than t he control. The apoptosis rate reached 24% after 48 h with the similar concentra tion of As2O3. Immunohistochemical study revealed that the expression of Bcl -2 was decreased after treated with As2O3. Conclusion: As 2O3 can depress the proliferation of SW1990 in vitro, mainly through the i nduction of apoptosis, and it is a potential agent for pancreatic cancer chemoth erapy.

Objective: To study the effect of arsenic trioxide (As 2O 3) on human pancreatic cancer cell proliferation and apoptosis (mainly early stage) in vitro . Methods: SW1990 cells line were treated with As 2O 3 at different concentration. Cell proliferation was evaluated by MTT and apoptosis by Annexin Ⅴ fluostaining, electron microscopy, flow cytometry and immunocytochemical staining of Bcl 2 and Bax. Results: As 2O 3 and cisplatin had the same cytotoxity on SW1990. The cytotoxic effect on tumor cell was produced by induction of apoptosis. Twelve hours after culture with 10 ?g/ml As 2O 3, much more SW1990 cells went into apoptosis than the control. The apoptosis rate reached 24% after 48 h with the similar concentration of As 2O 3. Immunohistochemical study revealed that the expression of Bcl 2 was decreased after treated with As 2O 3. Conclusion: As 2O 3 can depress the proliferation of SW1990 in vitro , mainly through the induction of apoptosis, and it is a potential agent for pancreatic cancer chemotherapy.

NKT细胞是一类特殊的T淋巴细胞亚群，既表达NK 细胞受体，也表达T细胞的相关受体。NKT细胞在多种免疫应答 的调节中发挥重要作用，包括感染、自身免疫性疾病、代谢性疾病和癌症，其通过连接固有免疫系统和适应性免疫系统显示出强大 的抗肿瘤活性。NKT细胞不仅能杀伤肿瘤细胞，也可激活其他抗肿瘤免疫细胞间接地发挥抗肿瘤作用，还能在肿瘤微环境中激活 衰竭的免疫细胞，在抗肿瘤免疫中发挥重要的作用。本文就NKT细胞的生物学特性及其在抗肿瘤免疫中的作用作一综述。

[Abstract] Objective: To investigate the short-term clinical efficacy and safety of intraperitoneal perfusion of natural killer (NK) cells in the treatment of ovarian cancer with ascites. Methods: The clinical data of 15 ovarian cancer patients with ascites effusion, who received NK cell perfusion in the Qinhuai Medical District of the General Hospital of Eastern Theater Command from November 2016 to January 2019, were analyzed. The peripheral blood was collected to isolate the peripheral blood mononuclear cells, and to further obtain the NK cells after culture. NK cell suspension was intraperitoneally perfused into the abdominal cavity (no less than 2×109 cells/ time). The volume of peritoneal effusion, the level of serum tumor marker CA-125, the level of serum cytokines IL-2, INF-γ and TNF-α as well as the changes in peripheral blood lymphocyte subsets were detected before and after the treatment; Moreover, the clinical efficacy and adverse reactions were observed. Results: The effective rate of intraperitoneal perfusion of NK cells was 66.7%, and there were no obvious treatment-related adverse reactions. Compared with before treatment, the serum tumor marker CA-125 level significantly decreased after treatment (P<0.05), and the levels of IL-15, IFN-γ and TNF-α increased significantly (P<0.05 or P<0.01), while there was no significant changes in peripheral blood lymphocyte subsets (all P>0.05). Conclusion: Intraperitoneal infusion of NK cells in the treatment of ovarian cancer associated peritoneal effusion has a good short-term clinical efficacy with little adverse reactions, which is a promising method for the treatment of cancerous peritoneal effusion.