AIM: To investigate the relationship betweenhepatocarcinogenesis and the expression of connexin32(cx32), connexin43 (cx43) mRNAs and proteins in vitro.METHODS Gap junction genes cx32 and cx43 mRNA inhepatocellular car...AIM: To investigate the relationship betweenhepatocarcinogenesis and the expression of connexin32(cx32), connexin43 (cx43) mRNAs and proteins in vitro.METHODS Gap junction genes cx32 and cx43 mRNA inhepatocellular carcinoma call lines HHCC, SMMC-7721 andnormal liver call line QZG were detected by in situhybridization (ISH) with digoxin-labeled cx32, and cx43cDNA probes. Expression of Cx32 and Cx43 proteins in thecell lines was revealed by indirect immuno-fluorescence andflow cytornetry (FCM).RESULTS Blue positive hybridization signals of cx32 andcx43 mRNAs detected by ISH with cx32 and cx43 cDNAprobes respectively were located in cytoplasm of cells ofHHCC, SMMC-7721 and QZG. No significant difference ofeither cx32 mRNA or cx43 mRNA was tested among HHCC,SMMC-7721 and QZG (P = 2.673, HHCC vs QZG; P =1.375, SMMC-7721 vs QZG). FCM assay showed that thepositive rates of Cx32 protein in HHCC, SMMC-7721 and QZGwere 0.7%, 1.7% and 99.0%, and the positive rates of Cx43protein in HHCC, SMMC-7721 and QZG were 7.3%, 26.5%and 99.1% respectively. Significant differences of both Cx32and Cx43 protein expression existed between hepatocellularcarcinoma cell lines and normal liver cell line ( P = 0.0069,HHCC vs QZG; P = 0.0087, SMMC-7721 vs QZG).Moreover, the fluorescent intensities of Cx32 and Cx43proteins in HHCC, SMMC-7721 were lower than that in QZG.CONCLUSION Hepatocellular carcinoma cell lines HHCCand SMMC-7721 exhibited lower positive rates andfluorescent intensities of Cx32, Cx43 proteins compared withthat of normal liver cell line QZG. lt is suggested that lowerexpression of both Cx32 and Cx43 proteins in hepatocellularcarcinoma cells could play pivotal roles in thehepatocarcinogenesis. Besides, genetic defects of cx32 andcx43 in post-translational processing should be considered.展开更多
AIM: To investigate gap junctional intercellular communication (GJIC) in hepatocellular carcinoma cell lines, and signal transduction mechanism of gap junction genes connexin32(cx32),connexin43(cx43) in human hepatoca...AIM: To investigate gap junctional intercellular communication (GJIC) in hepatocellular carcinoma cell lines, and signal transduction mechanism of gap junction genes connexin32(cx32),connexin43(cx43) in human hepatocarcinogenesis.METHODS: Scarped loading and dye transfer (SLDT) was employed with Lucifer Yellow (LY) to detect GJIC function in hepatocellular carcinoma cell lines HHCC, SMMC-7721and normal control liver cell line QZG. After Fluo-3AM loading, laser scanning confocal microscope (LSCM) was used to measure concentrations of intracellular calcium [Ca2+]i in the cells. The phosphorylation on tyrosine of connexin proteins was examined by immunoblot.RESULTS: SLDT showed that ability of GJIC function was higher in QZG cell than that in HHCC and SMMC-7721 cell lines. By laser scanning confocal microscopy, concentrations of intracellular free calcium [Ca2+]i was much higher in QZG cell line (108.37 nmol/L) than those in HHCC (35.13nmol/L) and SMMC-7721 (47.08 nmol/L) cells. Western blot suggested that only QZG cells had unphosphorylated tyrosine in Cx32 protein of 32 ku and Cx43 protein of 43ku; SMMC-7721 cells showed phosphorylated tyrosine Cx43 protein.CONCLUSION: The results indicated that carcinogenesis and development of human hepatocellular carcinoma related with the abnormal expression of cx genes and disorder of its signal transduction pathway, such as decrease of [Ca2+]i,post-translation phosphorylation on tyrosine of Cx proteins which led to a dramatic disruption of GJIC.展开更多
AIM: To explore the transcriptional gene expression profiles of HGF/SF-met signaling pathway in colorectal carcinoma to understand mechanisms of the signaling pathway at so gene level.METHODS: Total RNA was isolated f...AIM: To explore the transcriptional gene expression profiles of HGF/SF-met signaling pathway in colorectal carcinoma to understand mechanisms of the signaling pathway at so gene level.METHODS: Total RNA was isolated from human colorectal carcinoma cell line LoVo treated with HGF/SF (80 ng/L)for 48 h. Fluorescent probes were prepared from RNA labeled with cy3-dUTP for the control groups and with cy5-dUTP for the HGF/SF-treated groups through reversetranscription. The probes were mixed and hybridized on the microarray at 60 ℃ for 15-20 h, then the microarray was scanned by laser scanner (GenePix 4000B). The intensity of each spot and ratios of Cy5/Cy3 were analyzed and finally the differentially expressed genes were selected by GenePix Pro 3.0 software. 6 differential expression genes (3 up-regulated genes and 3 down-regulated genes) were selected randomly and analyzed by β-actin semiquantitative RT-PCR.RESULTS: The fluorescent intensities of built-in negative control spots were less than 200, and the fluorescent intensities of positive control spots were more than 5000.Of the 4004 human genes analyzed by microarray, 129 genes (holding 3.22 % of the investigated genes) revealed differential expression in HGF/SF-treated groups compared with the control groups, of which 61 genes were up-regulated (holding 1.52 % of the investigated genes) and 68 genes were down-regulated (holding 1.70 % of the investigated genes), which supplied abundant information about target genes of HGF/SF-met signaling.CONCLUSION: HGF/SF-met signaling may up-regulate oncogenes, signal transduction genes, apoptosis-related genes, metastasis related genes, and down-regulate a number of genes. The complexity of HGF/SF-met signaling to control the gene expression is revealed as a whole by the gene chip technology.展开更多
AIM:To study the effect of exposure to 8 MV X ray irradiation on heparanase mRNA expression in human colorectal carcinoma HT29 cells.METHOD:In situ hybridization technique was used to study the expression of heparanas...AIM:To study the effect of exposure to 8 MV X ray irradiation on heparanase mRNA expression in human colorectal carcinoma HT29 cells.METHOD:In situ hybridization technique was used to study the expression of heparanase mRNA in HT29 cells.RESULTS:After exposure to the dose of 2 Gy,the heparanase mRNA expression in HT29 cells was significantly weakened.CONCLUSION:Radiation may inhibit the metastasis ability of HT29 cells.展开更多
To provide the theoretical fundation for the further clinical application of mifepristone and anordrin compound. Materials & Methods Ribonuclease protection assay was used for the detection and q...To provide the theoretical fundation for the further clinical application of mifepristone and anordrin compound. Materials & Methods Ribonuclease protection assay was used for the detection and quantitation of estrogen and progesterone receptor mRNAs in human decidua from the termination of early pregnancy.Three groups, each of which had 6～8 cases, were studied. Results Compared to the normal control group, estrogen and progesterone receptor mRNAs increased significantly (P<0.05) in the mifepristone group, whereas the changes in the group administrated mifepristone compound which contains anordrin were not obvious. Conclusions The result suggests that with the similar clinical effect, mifepristone compound has less side effect on the patients, thus being more suitable for the anti early pregnancy drug.展开更多
Objective To explore the effects of mifepristone on the growth of human gastric cancer cell line MKN-45 and its possible mechanisms. Methods In situ hybridization was used to detect the expression of progesterone rece...Objective To explore the effects of mifepristone on the growth of human gastric cancer cell line MKN-45 and its possible mechanisms. Methods In situ hybridization was used to detect the expression of progesterone receptor (PR) mRNA in MKN-45 cells. Proliferation, cell cycle distribution, and the expression of Bcl-xL and vascular endothelial growth factor (VEGF) of MKN-45 cells incubated with various concentrations of mifepristone (1, 5, 10, and 20 μmol/L) were analyzed using MTT reduction assay, flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunoab-sorbent assay (ELISA), respectively. After transplantation of MKN-45 cells underneath the skin of athymic mice, mifepristone was administrated with the dose of 50 mg/(kg·d) for 6 weeks to evaluate the tumor growth. Apoptosis and the expression of proliferating cell nuclear antigen (PCNA) in xenografted tumors were detected using transmission electron microscopy and immunohistochemical staining, respectively. Results PR mRNA was highly expressed in cultured MKN-45 cell. Mifepristone dose-dependently inhibited the pr-oliferation of MKN-45 cells, and the inhibitory rate was dramatically increased from 7.21% to 47.23%. The inhibitory effect was accompanied by a dose-dependent increase in the percentage of cells in G 0 /G 1 phase, and with a concurrent decrease in the proportion of S- and G 2 /M-phase cells and the proliferative index from 57.65% to 24.54%. Meanwhile, mifepristone down-regulated the expression of Bcl-xL and VEGF in a dose-dependent manner. In vivo, mifepristone effectively inhibited the growth of xenografted tumors in nude mice (55.14% for inhibitory rate), induced apoptosis, and down-regulated PCNA expression in gastric cancer. Conclusion Mifepristone exerts significant growth inhibitory effects on PR-positive human MKN-45 gastric cancer cells via multiple mechanisms, and may be a beneficial agent against the tumor.展开更多
Objective To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1(MDR1) mRNA. Methods Two MDR1 cDNA vectors, pT7TS_MDR...Objective To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1(MDR1) mRNA. Methods Two MDR1 cDNA vectors, pT7TS_MDR1 and pGEM5Zf(+)_MDR1, were constructed and transcripted in vitro. Vector pGEM5Zf(+)_MDR1 only contained the coding region of mdr1 cDNA, and pT7TS_MDR1 also included Xeponus β-globin 5’ and 3’ untranslated region. MNCs were prepared from peripheral blood of parvicellular lung cancer patient. The two human mdr1 mRNAs were then transferred into human MNCs in vitro by DOTAP. And the expression efficiency and pump function of P-gp were measured with flow cytometry. Results Expression of P-gp significantly elevated in both transferred cells compared with untransferred cells (P < 0.01). And pT7TS_MDR1 showed higher capability in elevating the expression of P-gp than pGEM5Zf(+)_MDR1 (P < 0.01). The P-gp function was elevated in both pT7TS_MDR1 and pGEM5Zf(+)_MDR1 groups. The survival ratio of MNCs in erythrocyte-lysis-solution (ELS, 86.07%) and lymphocyte-isolation-solution (LIS, 83.67%) had no significant difference. The CD34+ cells content of the MNCs used for transfection was 2.65% and 1.01% in ELS and LIS group, respectively (P < 0.01).Conclusions It is a feasible approach to improve P-gp expression in human MNCs by transfection of MDR-1 mRNA. And the ELS may be more suitable for purifing MNCs for mRNA transfection than LIS.展开更多
Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles i...Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.展开更多
Objective To determine whether formation of the nucleolar channel system (NCS) in human endometrium depends on the presence of progesteronal steroids. Materials & Methods Tissues of late proliferative endometrium ...Objective To determine whether formation of the nucleolar channel system (NCS) in human endometrium depends on the presence of progesteronal steroids. Materials & Methods Tissues of late proliferative endometrium were obtained from 5 normally cycling women of reproductive age. Half of each tissue was cultured in the DMEM medium containing diethylstilbesterol (25 μg/mL) plus medroxyprogesterone acetate (25 μg/mL) (E + P culture). As a control, the other half was cultured in the medium alone. After 100 h incubation, the tissues were assessed for the formation of NCS with transmission electron microscope.Results NCS was observed in the endometrial epithelium treated with E + P or the medium alone. Moreover, giant mitochondria and glycogen accumulation were both seen in epithelia derived from both types of cultures.Conclusion Progesterone would be not indispensable for the formation of NCS in human endometrium. Transition of proliferative endometrium to the secretory stage in vitro could occur even in the absence of both estrogen and progesterone.展开更多
Objective To study the outcome of tension-free vaginal tape (TVT) for the treatment of stress urinary incontinence (SUI) in women with cystocele. Methods Forty-two patients with SUI confirmed by urodynamics underwent ...Objective To study the outcome of tension-free vaginal tape (TVT) for the treatment of stress urinary incontinence (SUI) in women with cystocele. Methods Forty-two patients with SUI confirmed by urodynamics underwent the TVT procedure under local anesthesia. A prolapse repair was done simultaneously. Results Mean TVT operation time was 26.29 minutes. Mean blood loss was 29.86 mL. Eighty-eight percent of the patients were able to micturate spontaneously within 12 hours and residual urine was less than 100 mL. And 12% of the patients had to use indwelling catheter for 3-11 days. Average hospital stay was 2.91 days. Eighty-eight percent of patients were discharged within 2 days. All patients were followed up (an average of 10.26 months). According to subjective and objective assessment of the outcome, 39 patients (93%) were cured, another 3 patients (7%) were significantly improved and none was failed. There were no major complications such as bladder injury occurred. Conclusion TVT procedure is a minimal invasive, effective, and safe surgery for treatment of SUI.展开更多
Thepathogenesisof prem ature ovarian failure (POF) rem ains unknow n. Accu- m ulated evidences indicate that abnorm ality in the im m une system m ay be one of the m ajor reasonsand theim balanceof Thelp typeI(Th1) ...Thepathogenesisof prem ature ovarian failure (POF) rem ains unknow n. Accu- m ulated evidences indicate that abnorm ality in the im m une system m ay be one of the m ajor reasonsand theim balanceof Thelp typeI(Th1) and Thelp typeII(Th2) m ay play an importantrolein theprocess. Study of thedistribution and functionalstatusof Th1/Th2 m ay behelpfulforevaluating thepathogenesisof POF.13 patientsw ith id- iopathic POFand 7 w om en of reproductiveagew ith norm alcyclew ereenrolled in this study.The percentage of Th1 and Th2 cellsfrom peripheralblood of thepatientsand the controls w as studied using FITC labelled CD4 m Ab to separate CD4+ cells by FACS, in com bination w ith in situ hybridization using Dig-labelled IL2, IL6 cDNA probes.Results:The patients w ith POF had significantly higher percentage of Th1 cellsascompared w ith the controls(49.76±9.22,20.06±7.10respectively, P< 0.001).The ratio of Th1/Th2 in thepatientsw ith POFw assignificantly higherthan thatof thecontrols(1.15±0.17, 0.63±0.09, P< 0.001). Conclusions: Thefindings of thisstudy suggestthatpatientsw ith POFhaveincreased Th1 cellactivation, w hich m ay be related to the pathogenesisof POF. Corresponding author: WANG Yi-li E-m ail: w yl@ irix. xam u. edu. cn展开更多
文摘AIM: To investigate the relationship betweenhepatocarcinogenesis and the expression of connexin32(cx32), connexin43 (cx43) mRNAs and proteins in vitro.METHODS Gap junction genes cx32 and cx43 mRNA inhepatocellular carcinoma call lines HHCC, SMMC-7721 andnormal liver call line QZG were detected by in situhybridization (ISH) with digoxin-labeled cx32, and cx43cDNA probes. Expression of Cx32 and Cx43 proteins in thecell lines was revealed by indirect immuno-fluorescence andflow cytornetry (FCM).RESULTS Blue positive hybridization signals of cx32 andcx43 mRNAs detected by ISH with cx32 and cx43 cDNAprobes respectively were located in cytoplasm of cells ofHHCC, SMMC-7721 and QZG. No significant difference ofeither cx32 mRNA or cx43 mRNA was tested among HHCC,SMMC-7721 and QZG (P = 2.673, HHCC vs QZG; P =1.375, SMMC-7721 vs QZG). FCM assay showed that thepositive rates of Cx32 protein in HHCC, SMMC-7721 and QZGwere 0.7%, 1.7% and 99.0%, and the positive rates of Cx43protein in HHCC, SMMC-7721 and QZG were 7.3%, 26.5%and 99.1% respectively. Significant differences of both Cx32and Cx43 protein expression existed between hepatocellularcarcinoma cell lines and normal liver cell line ( P = 0.0069,HHCC vs QZG; P = 0.0087, SMMC-7721 vs QZG).Moreover, the fluorescent intensities of Cx32 and Cx43proteins in HHCC, SMMC-7721 were lower than that in QZG.CONCLUSION Hepatocellular carcinoma cell lines HHCCand SMMC-7721 exhibited lower positive rates andfluorescent intensities of Cx32, Cx43 proteins compared withthat of normal liver cell line QZG. lt is suggested that lowerexpression of both Cx32 and Cx43 proteins in hepatocellularcarcinoma cells could play pivotal roles in thehepatocarcinogenesis. Besides, genetic defects of cx32 andcx43 in post-translational processing should be considered.
文摘AIM: To investigate gap junctional intercellular communication (GJIC) in hepatocellular carcinoma cell lines, and signal transduction mechanism of gap junction genes connexin32(cx32),connexin43(cx43) in human hepatocarcinogenesis.METHODS: Scarped loading and dye transfer (SLDT) was employed with Lucifer Yellow (LY) to detect GJIC function in hepatocellular carcinoma cell lines HHCC, SMMC-7721and normal control liver cell line QZG. After Fluo-3AM loading, laser scanning confocal microscope (LSCM) was used to measure concentrations of intracellular calcium [Ca2+]i in the cells. The phosphorylation on tyrosine of connexin proteins was examined by immunoblot.RESULTS: SLDT showed that ability of GJIC function was higher in QZG cell than that in HHCC and SMMC-7721 cell lines. By laser scanning confocal microscopy, concentrations of intracellular free calcium [Ca2+]i was much higher in QZG cell line (108.37 nmol/L) than those in HHCC (35.13nmol/L) and SMMC-7721 (47.08 nmol/L) cells. Western blot suggested that only QZG cells had unphosphorylated tyrosine in Cx32 protein of 32 ku and Cx43 protein of 43ku; SMMC-7721 cells showed phosphorylated tyrosine Cx43 protein.CONCLUSION: The results indicated that carcinogenesis and development of human hepatocellular carcinoma related with the abnormal expression of cx genes and disorder of its signal transduction pathway, such as decrease of [Ca2+]i,post-translation phosphorylation on tyrosine of Cx proteins which led to a dramatic disruption of GJIC.
基金National Natural Science-Foundation of China,No.30170486 and No.30170423
文摘AIM: To explore the transcriptional gene expression profiles of HGF/SF-met signaling pathway in colorectal carcinoma to understand mechanisms of the signaling pathway at so gene level.METHODS: Total RNA was isolated from human colorectal carcinoma cell line LoVo treated with HGF/SF (80 ng/L)for 48 h. Fluorescent probes were prepared from RNA labeled with cy3-dUTP for the control groups and with cy5-dUTP for the HGF/SF-treated groups through reversetranscription. The probes were mixed and hybridized on the microarray at 60 ℃ for 15-20 h, then the microarray was scanned by laser scanner (GenePix 4000B). The intensity of each spot and ratios of Cy5/Cy3 were analyzed and finally the differentially expressed genes were selected by GenePix Pro 3.0 software. 6 differential expression genes (3 up-regulated genes and 3 down-regulated genes) were selected randomly and analyzed by β-actin semiquantitative RT-PCR.RESULTS: The fluorescent intensities of built-in negative control spots were less than 200, and the fluorescent intensities of positive control spots were more than 5000.Of the 4004 human genes analyzed by microarray, 129 genes (holding 3.22 % of the investigated genes) revealed differential expression in HGF/SF-treated groups compared with the control groups, of which 61 genes were up-regulated (holding 1.52 % of the investigated genes) and 68 genes were down-regulated (holding 1.70 % of the investigated genes), which supplied abundant information about target genes of HGF/SF-met signaling.CONCLUSION: HGF/SF-met signaling may up-regulate oncogenes, signal transduction genes, apoptosis-related genes, metastasis related genes, and down-regulate a number of genes. The complexity of HGF/SF-met signaling to control the gene expression is revealed as a whole by the gene chip technology.
文摘AIM:To study the effect of exposure to 8 MV X ray irradiation on heparanase mRNA expression in human colorectal carcinoma HT29 cells.METHOD:In situ hybridization technique was used to study the expression of heparanase mRNA in HT29 cells.RESULTS:After exposure to the dose of 2 Gy,the heparanase mRNA expression in HT29 cells was significantly weakened.CONCLUSION:Radiation may inhibit the metastasis ability of HT29 cells.
基金This work was supported by a grant from the National Laboratory of Contraceptives and Devices Research of China
文摘To provide the theoretical fundation for the further clinical application of mifepristone and anordrin compound. Materials & Methods Ribonuclease protection assay was used for the detection and quantitation of estrogen and progesterone receptor mRNAs in human decidua from the termination of early pregnancy.Three groups, each of which had 6～8 cases, were studied. Results Compared to the normal control group, estrogen and progesterone receptor mRNAs increased significantly (P<0.05) in the mifepristone group, whereas the changes in the group administrated mifepristone compound which contains anordrin were not obvious. Conclusions The result suggests that with the similar clinical effect, mifepristone compound has less side effect on the patients, thus being more suitable for the anti early pregnancy drug.
文摘Objective To explore the effects of mifepristone on the growth of human gastric cancer cell line MKN-45 and its possible mechanisms. Methods In situ hybridization was used to detect the expression of progesterone receptor (PR) mRNA in MKN-45 cells. Proliferation, cell cycle distribution, and the expression of Bcl-xL and vascular endothelial growth factor (VEGF) of MKN-45 cells incubated with various concentrations of mifepristone (1, 5, 10, and 20 μmol/L) were analyzed using MTT reduction assay, flow cytometry, reverse transcription-polymerase chain reaction (RT-PCR), and enzyme-linked immunoab-sorbent assay (ELISA), respectively. After transplantation of MKN-45 cells underneath the skin of athymic mice, mifepristone was administrated with the dose of 50 mg/(kg·d) for 6 weeks to evaluate the tumor growth. Apoptosis and the expression of proliferating cell nuclear antigen (PCNA) in xenografted tumors were detected using transmission electron microscopy and immunohistochemical staining, respectively. Results PR mRNA was highly expressed in cultured MKN-45 cell. Mifepristone dose-dependently inhibited the pr-oliferation of MKN-45 cells, and the inhibitory rate was dramatically increased from 7.21% to 47.23%. The inhibitory effect was accompanied by a dose-dependent increase in the percentage of cells in G 0 /G 1 phase, and with a concurrent decrease in the proportion of S- and G 2 /M-phase cells and the proliferative index from 57.65% to 24.54%. Meanwhile, mifepristone down-regulated the expression of Bcl-xL and VEGF in a dose-dependent manner. In vivo, mifepristone effectively inhibited the growth of xenografted tumors in nude mice (55.14% for inhibitory rate), induced apoptosis, and down-regulated PCNA expression in gastric cancer. Conclusion Mifepristone exerts significant growth inhibitory effects on PR-positive human MKN-45 gastric cancer cells via multiple mechanisms, and may be a beneficial agent against the tumor.
文摘Objective To evaluate the expression and function activity of P-glycoprotein (P-gp) in human mononuclear cells (MNCs) in vitro transfected by multidrug resistance-1(MDR1) mRNA. Methods Two MDR1 cDNA vectors, pT7TS_MDR1 and pGEM5Zf(+)_MDR1, were constructed and transcripted in vitro. Vector pGEM5Zf(+)_MDR1 only contained the coding region of mdr1 cDNA, and pT7TS_MDR1 also included Xeponus β-globin 5’ and 3’ untranslated region. MNCs were prepared from peripheral blood of parvicellular lung cancer patient. The two human mdr1 mRNAs were then transferred into human MNCs in vitro by DOTAP. And the expression efficiency and pump function of P-gp were measured with flow cytometry. Results Expression of P-gp significantly elevated in both transferred cells compared with untransferred cells (P < 0.01). And pT7TS_MDR1 showed higher capability in elevating the expression of P-gp than pGEM5Zf(+)_MDR1 (P < 0.01). The P-gp function was elevated in both pT7TS_MDR1 and pGEM5Zf(+)_MDR1 groups. The survival ratio of MNCs in erythrocyte-lysis-solution (ELS, 86.07%) and lymphocyte-isolation-solution (LIS, 83.67%) had no significant difference. The CD34+ cells content of the MNCs used for transfection was 2.65% and 1.01% in ELS and LIS group, respectively (P < 0.01).Conclusions It is a feasible approach to improve P-gp expression in human MNCs by transfection of MDR-1 mRNA. And the ELS may be more suitable for purifing MNCs for mRNA transfection than LIS.
文摘Aim: To evaluate the fertilization competence of spermatozoa from ejaculates and testicle when the oocytes were matured in vitro following intracytoplasmic sperm injection (ICSI). Methods: Fifty-six completed cycles in 46 women with polycystic ovarian syndrome were grouped according to the semen parameters of their male partners. Group 1 was 47 cycles that presented motile and normal morphology spermatozoa in ejaculates and Group 2 was the other nine cycles where male partners were diagnosed as obstructive azoospermia and spermatozoa could only be found in testicular tissue fragment. All female patients received minimal stimulation with gonadotropin. Immature oocytes were matured in vitro and inseminated by ICSI. The spermatozoa from testes were retrieved by testicular fine needle aspiration. Results: A total of 449 and 78 immature oocytes were collected and cultured for 48 hours, 75.5 % (339/449) and 84.6 % (66/78) oocytes were matured in Groups 1 and 2, respectively. The percentage of oocytes achieving normal fertilization was significantly higher in Group 1 than that in Group 2 (72.9 % vs. 54.5 %, P < 0.05). There were no significant differences in the rates of oocytes cleavage and clinical pregnancies in these two groups [87.4 % (216/247) vs. 88.9 % (32/36); 21.3 % (10/47) vs. 44.4 % (4/9)]. A total of 15 babies in the two groups were healthy delivered at term. Conclusion: It appears that IVM combined with ICSI using testicular spermatozoa can produce healthy infants, while the normal fertilization rate of in vitro matured oocytes after ICSI using testicular spermatozoa was significantly lower than using the ejaculated spermatozoa.
基金This study was supported by the National Science Fund of P.R.China (No.39970765)
文摘Objective To determine whether formation of the nucleolar channel system (NCS) in human endometrium depends on the presence of progesteronal steroids. Materials & Methods Tissues of late proliferative endometrium were obtained from 5 normally cycling women of reproductive age. Half of each tissue was cultured in the DMEM medium containing diethylstilbesterol (25 μg/mL) plus medroxyprogesterone acetate (25 μg/mL) (E + P culture). As a control, the other half was cultured in the medium alone. After 100 h incubation, the tissues were assessed for the formation of NCS with transmission electron microscope.Results NCS was observed in the endometrial epithelium treated with E + P or the medium alone. Moreover, giant mitochondria and glycogen accumulation were both seen in epithelia derived from both types of cultures.Conclusion Progesterone would be not indispensable for the formation of NCS in human endometrium. Transition of proliferative endometrium to the secretory stage in vitro could occur even in the absence of both estrogen and progesterone.
文摘Objective To study the outcome of tension-free vaginal tape (TVT) for the treatment of stress urinary incontinence (SUI) in women with cystocele. Methods Forty-two patients with SUI confirmed by urodynamics underwent the TVT procedure under local anesthesia. A prolapse repair was done simultaneously. Results Mean TVT operation time was 26.29 minutes. Mean blood loss was 29.86 mL. Eighty-eight percent of the patients were able to micturate spontaneously within 12 hours and residual urine was less than 100 mL. And 12% of the patients had to use indwelling catheter for 3-11 days. Average hospital stay was 2.91 days. Eighty-eight percent of patients were discharged within 2 days. All patients were followed up (an average of 10.26 months). According to subjective and objective assessment of the outcome, 39 patients (93%) were cured, another 3 patients (7%) were significantly improved and none was failed. There were no major complications such as bladder injury occurred. Conclusion TVT procedure is a minimal invasive, effective, and safe surgery for treatment of SUI.
文摘Thepathogenesisof prem ature ovarian failure (POF) rem ains unknow n. Accu- m ulated evidences indicate that abnorm ality in the im m une system m ay be one of the m ajor reasonsand theim balanceof Thelp typeI(Th1) and Thelp typeII(Th2) m ay play an importantrolein theprocess. Study of thedistribution and functionalstatusof Th1/Th2 m ay behelpfulforevaluating thepathogenesisof POF.13 patientsw ith id- iopathic POFand 7 w om en of reproductiveagew ith norm alcyclew ereenrolled in this study.The percentage of Th1 and Th2 cellsfrom peripheralblood of thepatientsand the controls w as studied using FITC labelled CD4 m Ab to separate CD4+ cells by FACS, in com bination w ith in situ hybridization using Dig-labelled IL2, IL6 cDNA probes.Results:The patients w ith POF had significantly higher percentage of Th1 cellsascompared w ith the controls(49.76±9.22,20.06±7.10respectively, P< 0.001).The ratio of Th1/Th2 in thepatientsw ith POFw assignificantly higherthan thatof thecontrols(1.15±0.17, 0.63±0.09, P< 0.001). Conclusions: Thefindings of thisstudy suggestthatpatientsw ith POFhaveincreased Th1 cellactivation, w hich m ay be related to the pathogenesisof POF. Corresponding author: WANG Yi-li E-m ail: w yl@ irix. xam u. edu. cn
