Objective To evaluate the treatment and apoptosis effect of adenovirus-mediated herpes simplex virus thymidine kinase (HSV 1-tk) gene transference followed by administration of ganciclovir (GCV) and acyclovir (ACV) on ovarian epithelial cancer cells. Methods Recombinant adenovirus was amplificated and purified by routine method. The expression of HSV 1-tk gene was assayed by polymerase chain reaction (PCR). The efficiency of recombinant Advtk transference was evaluated. The cytotoxicity efficacy of TYK cells that carry HSV 1-tk gene was evaluated followed by GCV、ACV administration after the transference of Advtk. The changes and apoptosis of TYK cells that carry HSV 1-tk gene were observed by means of analysis of DNA fragmentation and electronic microscopy. Results Adenovirus was amplificated and purified in large amount. PCR assay showed 404 bp special band. When the multiplicities of infection (MOI) was 100, the transduction rate was 98 9%. The inhibition rates of TYK cells that carry HSV 1-tk gene increased with the increase of MOI when the same concentration of GCV、 ACV were given. When the MOI was same, the inhibition rates were also increased with the increase concentration of GCV、ACV. Apoptosis of TYK cells that carry HSV 1-tk gene after administration of GCV was observed by means of analysis of DNA fragmentation and electronic microscopy. Conclusions The HSV 1-tk gene can effectively transfered into TYK cells by recombinant replicated-deficient adenovirus vector, GCV、ACV can effectively kill TYK cells that carry HSV 1-tk gene in vitro. Apoptosis may be the mechanism of the killing effect.

Objective To investigate the proliferation and cytotoxicity of cytotoxic T lymphocyte (CTL) induced by ovarian cancer cells transfected with a CD 80 gene containing retroviral vector Methods The ovarian cancer cell line TYK cells were transfected with retro virus vector PLXSN hCD 80 After geneticin (G418) selection, the CD 80 expression of the transfectants was confirmed by flow cytometry CTL was induced by co culture of CD 80 expressing TYK cells (TYK hCD 80 ) and peripheral blood mononuclear cells (PBMC) in the presence of anti CD 3 monoclonal antibody (McAb) Proliferation of PBMC was measured using 3?H Thymidine incorporation assay The lysis activity of CTL toward tumor cells was determined using methyl thiazolyl tetrazolium (MTT) assay Results After transfection and G418 selection, the CD 80 expression was proved by flow cytometry The highest rate of CD 80 expression was 84 9% The TYK cell line expressing high CD 80 was named TYK hCD 80 In the presence of anti CD 3 McAb,TYK hCD 80 significantly enhanced proliferation of PBMC than TYK cells ( 3H Thymidine incorporation, (40 604? 842) vs (8 000? 594) cpm ( P

Objective:To investigate the expression of NKG2D in peripheral blood of patients with ovarian benign or malignant tumors, and the expression of the human MHC class Ⅰ chain-related gene A(MICA) on the correspondent tumor tissues. To analyze and discuss the function of NKG2D in anti-ovarian cancer mechanism and its relation with immune escaping of cancer.Methods:Flow cytometry analysis was used to detect NKG2D in the peripheral blood of 42 ovarian carcinoma patients, 23 ovarian tumor patients and 20 health persons. The expressions of MICA in part of the correspondent tissues were examined by means of reverse transcription-polymerase chain reaction (RT-PCR).Results:The expression of NKG2D were (94 23?6 02)%?(98 70?0 98)%?(98 61?1 59)% respectively, compared with the other two groups, the NKG2D expression in malignant group was significantly low; The rate of MICA mRNA expression in ovarian carcinomas was significantly higher than that in benign and normal tissues. No evident relation was found between the expression of MICA mRNA and the clinical factors.Conclusion:The activity of NK cell and the anti-cancer cellular immunity level reduce in patients with ovarian cancer; The decrease of the receptor NKG2D is a reason for the descend of the activity of NK cells; MICA mRNA expression is relative to malignant transformation; the immune-escape of ovarian cancer probably is relative to the down-regulation of NKG2D and the up-regulation of its ligand MICA.

Purpose:To study the effects of arsenic trioxide on the different types of human ovarian cancer cells growth in vitro.Methods:Methyl thiazolyl tetrazolium (MTT)was used to observe the growth inhibition rates of human ovarian cancer cell lines 3AO,SKOV 3 and TYK cells by various concentration arsenic trioxide at different times; Cell apoptosis percentage and cell cycles phase distribution of 3AO were measured by flow cytometry(FCM) assays; Apoptotic phenotype of SKOV 3 was observed by acridine dying. Results:Arsenic trioxide could inhibit the growth of 3AO?SKOV 3 and TYK effectively, depended on the action time and concentration of the medicine(P0.05). Within a certain range, 3AO cells apoptotic percentage induced by arsenic trioxide were enhanced in concentration- and time-dependent patterns(P

Objective: To observe the synergistic inhibitory effect of angiostatin gene combined with antisense hypoxia inducible factor-1? (aHIF-1?) gene on implanted human ovarian carcinoma in nude mice. Methods: BALB/C nude mice were subcutaneously transplanted with SKOV3 tumor cells and the tumors were allowed to grow till the diameter reached 0.4 cm, then the mice were randomly divided into 4 groups: PcDNA3 control group, PcDNA3-Angiostatin group, PcDNA3B-aHIF-1? group and PcDNA3-Angiostatin+PcDNA3B-aHIF-1? group; plamids PcDNA3, PcDNA3-Angiostatin, PcDNA3B-aHIF-1? and PcDNA3-Angiostatin+PcDNA3B-aHIF-1? were injected intra-tumorally in the above groups, respectively. The tumor samples were harvested on the 7 th day after gene transfer. Angiostatin, HIF-1?, vascular endothelial growth factor (VEGF) and microvessel density (MVD) of tumors were determined by immunohistochemical methods. Tumor cell apoptosis was determined with TUNEL staining. Results:The growth of tumors of PcDNA3-Angiostatin+PcDNA3B-aHIF-1? group was significantly inhibited, with local low expression of HIF-1? and VEGF (lower than those of the other 3 groups). MVD in combined transfection group(13.6?2.3) was lower than that of Angiostain group (24.5?2.7); the apoptosis index in combined transfection group (5.32?0.62)was higher than those of Angiostatin group(2.89?0.45), aHIF-1? group(2.98?0.51)and contrl group(1.56?0.41). Conclusion: Our results suggest a synergestic effect between Angiostain gene and aHIF-1? gene in inhibiting implanted human ovarian tumors in nude mice, which may contribute to drug resistance in antiangiogenic therapy of tumors.

Objective To investigate the targeted killing effect of mucin 1 single chain antibody targeting, lentivirus mediated suicide gene therapy and ganciclovir(GCV) in mucin 1 + ovarian epithelial carcinoma cells Methods Mucin 1 single chain antibody targeting lentivirus produced by packaging cell line 293T transduced herpes simplex virus thymidine kinase (HSV tk) gene into the ovarian epithelial cancer cell line SKOV3 (MUC1 +) The infection effect was observed through fluorescence microscopy Polymerase chain reaction (PCR) and reverse transcription PCR (RT PCR) were resorted to demonstrate the successful transduction and transcription of the HSV tk gene After administration of GCV, changes of those cells were observed through optical microscopy The cytotoxicity efficacy of HSV tk/GCV system was evaluated by methyl thiazolyl tetrazolium method Results It was observed through fluorescence microscopy that anti MUC1 directed lentivirus can specificly infect MUC1 + ovarian cancer cells A fragment of 600 bp was generated through PCR and RT PCR which indicated successful transduction and transcription of the HSV tk gene in SKOV3 cells Changes of cells followed by administration of GCV were observed with optical microscopy Significant cytotoxicity efficacy of GCV to SKOV3 was observed Conclusions The HSV tk gene can be targetedly transducted into MUC1 + ovarian cancer cell line under the mediation of anti MUC1 directed lentivirus, and such HSV tk/GCV system has targetingly killing effect on MUC1 + cancer cells

Objective To explore the expression and significance of chemokine CXC reeeptor (CXCR)3 and CXCR4 and their ligands(CXCL)at the early pregnancy decidua and villi.Methods Decidual mononuclear cells were isolated from the normal decidua of 5-8 weeks pregnant women by lymphocyte separation medium in vitro.CD56+natural killer(NK)cells were purified by dynabeads cell sorter kiL Purity and phenotype of CD56+decidua NK cells were analyzed by fluorescence-activated eell sorter (FACS).Gene expression of CXCR3 and CXCR4 in decidua NK cells and CXCL9,CXCL10 and CXCL12 in early pregnancy decidua and villi was assessed bv RT.PCIZ Protein expression of CXCL9,CXCL10 in normal endometrium and early pregnancy decidua was characterized and quantified by streptavidin-biotin pemxidase chain reaction(SP)immunohistochemistry and computered image analysis system.Correlations between the gray degree of CXCL9 and CXCL10 and the number of CD56+NK cells in upper tissue were analyzed by Spearman's correlation ceefficient rank tesL Results The phenotype of 98.7%decidua NK cells was CD56bright.The genes of CXCR3 and CXCR4 were expressed in decidua NK cells and that of CXCL9 and CXCL1O were expressed in early pregnancy decidua and CXCLI2 in early pregnancy villi.CXCL9 and CXCL10 were expressed in the cytoplasm of surface epithelia,glandular epithelia and stromal cells of early pregnancy deeidua and were not expressed in villi by immunohistochemistry.The gray degree of CXCL9 and CXCL10 in the secretory phase endometrium(56±43,59±47)was stronger than that in the proliferative phase(16±18,8±14,P<0.05)and reached the highest(143±35,158±29,P<0.05)in the early pregnancy decidua.The number of cD+56 NK cell in the secretory phase endometrium(60±20)was more than that in the proliferative phase endometrium(23±4,P<0.05)and was the most in the early pregnancy decidua(114±15,P<0.05).The gray degree of CXCL9 in upper tissue had a positive correlation with the number of CD+56 cells(r=0.88,P<0.05)and that of CXCL10 had a similar pattern to CXCL9(r=0.86,P<0.05).Condusion The interactions between CXCL9,CXCL10 and CXCL12 expressed in decidua and villi and CXCR3,CXCR4 expressed in CD+56 decidua NK cells may influence the CD+56 NK cell recruitment at the maternal-fetal interface.

Objective To explore the effects of ursodeoxycholic acid (UDCA) on the fluidity of rat visceral sac and placental glutathione (GSH) concentration in rats with intrahepatic cholestasis. Methods Sixty rats were randomly divided into 3 groups (20 in each). Refined vegetable oil 2.5 ml/(kg?d) was given to the control group since the 13 days of pregnancy. The ICP treatment and non treatment group received either progesterone 75 mg/(kg?d) or 17? ethynylestradiol 1.25 mg/ (kg?d) from the 13th to 17th day, respectively. From the 17th day, the control and non treatment group were fed with 0.9% nitrachloride solution 5 mg/(kg?d) and the treatment group with UDCA 50 mg/(kg?d). All rats were sacrificed on the 21st day. The visceral yolk sac cell membrane and GSH concentration were measured Results The concentration of GSH in the ICP non treatment group (1.12?0.02 mmol/g protein) was significantly lower than that of the treatmentgroup (1.38?0.03 mmol/g protein) and the control group (1.56?0.07 mmol/g protein) ( P 0.05). The fetal death rate in treatment group (9.55%) and control group (1.97%) was significantly lower than that of the non treatment group (20.47%) ( P

Objective To investigate the influences of active immunotherapy on T helper cell (Th)1/Th2 type cytokines in women with unexplained habitual abortion(UHA) Methods A total of 55 patients with UHA were studied, including 30 cases after active immunotheraphy (AIT) and 25 cases without any therapy(NAIT). Fifteen cases of normal nonpregnant (NNP) women were selected as control group. Supernatants from trophoblast activated peripheral blood mononuclear cells (PBMC) of the three groups were tested by enzyme linked immunosorbent assay (ELISA) for interferon gamma (IFN ?), interleukin 2 (IL 2), IL 4, IL 10. Results (1) The levels of IL 2 and IFN ? in AIT group [(108?37) ng/L and (110 ?52) ng/L, respectively] were lower significantly than those in NAIT group[(223?85) ng/L and (326 ?92) ng/L, respectively]( P 0.05). (2) Twenty six women in AIT group got pregnant, but 8 women experienced pregnancy loss repeatedly whose IL 2?IFN ? levels were higher than those in other 18 women got successful pregnancy and IL 4,IL 10 levels lower than the latter. Conclusions UHA patients have Th1 type immunity to trophoblast and produce high level Th1 type cytokines which probably result in pregnancy loss. Active immunotheraphy could make a shift from Th1 to Th2 immunity, thus favoring the maintenance of pregnancy.

Objective:To study the cooperated effect of Human Embryonic Lung Fibroblast(HELF) feeder layer and LIF in the culture of human Embryonic Stem(hES) cells and establish the method of identifying hES cells from Primordial Germ Cells(PGCs).Methods:Embryonic lungs were mechanically disaggregated,then cultured to establish HELF feeder layer.Gonadal ridges and mesenteries of embryos were mechanically disaggeregated,then cultured and passaged in vitro.Comparing the growth characters of hES cells in different conditions.To identify hES cells through their biological characteristics.Results:The coactions of HELF feeder layer and LIF play an important role in proliferation and undifferentiation of hES cells in vitro.High levels of AKP and telomerase activity are associated with hES cells. The cultrued cells have been continuously passaged for more than two months and found to be karyotypically normal and stable.When differentiating,they form embryoid bodies(EBs).Conclusion:To avoid indection of the heterogeneous protein,HELF feeder layer, in the presence of LIF,can be used in culture of hES cells;hES cells from PGCs can be identified through their biological characteristic. [