1.Expression of connexin 43 in ovarian cancer and its relationship with chemoresistance
Chinese Journal of Obstetrics and Gynecology 2009;44(1):50-55
Objective To examine the expression of protein kinase C (PKC), connexin43(Cx43) and non-phosphorylated Cx43 in ovarian cancer, and discuss the role of phosphorylated of Cx43 in chemoresistance in ovarian cancer. Methods We examined the expression of Cx43, non-phosphorylated Cx43 and PKC in ovarian cancer tissue by immunohistochemistry, and compared their expression in chemosensitivity group and ehemoresistance group. Cisplatin resistant ovarian cancer cell line SKOV3/DDP cells were treated by staurosporine (a kind of PKC inhibitors). Then expression of Cx43, non-phosphorylated Cx43 and PKC were tested. Meanwhile, we tested chemosensitivity of SKOV3/DDP cells by ATP bioluminescence tumor chemosensifivity assay (ATP-TCA). Results (1) Immunohistochemically,the rates of positive expression of Cx43 and non-phospharylated Cx43 were 54%, 14% respectively in the chemoresistance group, which were 83%, 59% in the chemosensitivity group respectively (P<0.05). The rate of positive expression of PKC in 28 chemoresistance ovarian cancer cases (64%) was higher than that in 29 chemosensitivity cases (31% ,P<0.05). Both of them were significantly lower in ehemoresistanee group than in chemosensitivity group (P<0.05). In addition, the expression of PKC was negatively correlated with the expression of Cx43 and non-phosphorylated Cx43. The correlation coefficients were -0. 626 and -0. 714, respectively (P<0.05). (2) Immunohistochemically, PKC was down regulated, and Cx43 and non-phosphorylated Cx43 were up regulated in SKOV3/DDP cells after staurosporine treatment. The longer the staurosporine worked, the more expression of Cx43 was. (3) By ATP-TCA, SKOV3/DDP cells were resistant to paclitaxel and cisplatin. The tumor growth inhibition was higher in the group of paclitaxel or cisplatin combined staurosporine than in the group of paclitaxel or cisplatin alone. The sensitivity was intermediate in the group combined with low concentration staurosporine (1×10-8 moL/L), and the sensitivity was high in the group combined with high concentration staurosporine (1×10-7 mol/L). Conclusions Phosphorylation of Cx43 caused by PKC leads to decrease in the expression of Cx43. This effect makes ovarian cancer cells less chemosensitive. Phosphorylation of Cx43 caused by PKC can be inhibited by staurosporine.
2.Expression and function of cyclooxygenase-2 in endometrial carcinoma
Journal of Peking University(Health Sciences) 2009;41(6):657-663
Objective: To evaluate the cyclooxygenase-2 ( COX-2) expression at mRNA and protein levels , as well as its clinical significance, and to study the correlation of COX-2 with angiogenesis, apoptosis and estrogen or progestron receptors. Methods: Forty-one samples of normal endoraetrium tissues and 52 ones of endometrial carcinomas( EC) were collected,together with their corresponding clinical information. RT-PCR was adopted to determine the expression of COX-2 mRNA. Immunohistochemical staining was engaged to evaluate the protein expression of COX-2, VEGF, Bcl-2 and the MVD marked by CD105. And TUNEL was used to count the apoptosis cells. Results: (1) Significantly higher mRNA and protein levels of COX-2 were found in endometrial carcinomas than in normal endometrial tissues ( P<0.001) . (2) At protein level ,COX-2 expression was related to the grade(P =0.020) , but had nothing to do with the stage, pathological type, depth of myometrium invasion and lymph node metastasis. Neither did the level of COX-2 mRNA with all the factors above. (3)The level of COX-2 mRAN had nothing to do with that of protein(P =0. 125, r =0. 222). (4) The higher the expression of COX-2 protein was, the higher that of VEGF,MVD or Bcl-2 was,and the lower the count of apoptosis cells was P<0.05. (6)The upregulation of COX-2 protein had no correlation with ER-α and ER-β . But it had positive correlation with PRA and PRB,P was 0.031 and 0.007 .respectively. Conclusion: (1)The expression of COX-2 is significantly higher in EC than in normal endometrium tissue, not only at the mRNA level, but also at protein level. ( 2 ) At protein level, COX-2 expression is related to the grade, but has nothing to do with the stage, pathological type, depth of myometrium invasion and lymph node metastasis. Neither does the level of COX-2 mRNA with all the factors above. (3)The upregulation of COX-2 protein in EC correlates with the increase of VEGF protein and MVD. (4) The upregulation of COX-2 protein in EC correlates with the increase of Bcl-2 protein and the decrease of apoptosis cells. (5)The upregulation of COX-2 protein in EC has no significant correlation with ER expression, but has something to do with the increase of PRA and PRB.
3.Re-evaluation on perinatal screening of TORCH
Chinese Journal of Laboratory Medicine 2008;31(7):742-746
TORCH infection during pregnancy is a group of infectious diseases resulting from viruses and other microorganisms. According to the past over 20-year experiences in serum screening in China, several infections which should be performed in the first trimester, or even much earlier in pregnant period, are hepatitis B, syphilis, HIV, and these three infections are confirmed to be harmful to the fetus or infants and also diagnostic methods and treatment are available. Screening for rubella should be better performed before conception and no specific management is required with IgM+, while immunization is recommended before pregnancy with IgM-. Screenings for cytomegalovirus, toxoplasmosis and herpes virus are not recommended. Scnography is warranted if fetal abnormalities were resulted from the above infections and amniocentesis or cordocentesis might be required for further investigations.
4.Isolation and culturing of highly purified glandular epithelial cells from human endometrial specimens
Journal of Medical Postgraduates 2003;0(05):-
Objective: To develop an improved method for the primary isolation and culture of highly purified endometrial glandular epithelial cells in vitro.Methods: Normal endometrial tissues were isolated and cultured in vitro by double collagenase digestion and selective wall-pasting.Results: Highly purified endometrial epithelial cells were successfully isolated and cultured.Conclusion: Epithelial and interstitial cells of the endometrium can be successfully isolated with high purity by double collagenase digestion and selective wall-pasting.
5.Role of human leukocyte antigen-G small interference RNA in a choriocarcinoma cell line
Chinese Journal of Obstetrics and Gynecology 2001;0(08):-
Objective To study the role of human leukocyte antigen-G (HLA-G) in the carcinogenesis and development of choriocarcinoma. Methods We designed and synthesized a double strand small interference RNA (siRNA) of HLA-G, then transfected it into a HLA-G overexpressed choriocarcinoma cell line, JEG-3 by lipofectine; HLA-G mRNA level was detected by real time RT-PCR; HLA-G protein level was detected by western blot. The living cells of JEG-3 were counted under microscope after transfection by siRNA. Results Double strand siRNA of HLA-G effectively downregulated the mRNA level and the protein level of HLA-G. The mRNA levels by Ct value at different concentration (1.0, 2.5, 5.0 ?g/L) were 20.67?0.02, 21.37?0.03,21.43?0.02, respectively. They were significantly different compared with the control groups which were 20.33?0.01, 20.37?0.02,20.40?0.03, respectively ( P
6.Microecological investigation of vaginal microflora in women with varying degree gynecologic symptoms in clinics
Bingbing XIAO ; Zhaohui LIU ; Qinping LIAO
Chinese Journal of Obstetrics and Gynecology 2009;44(1):6-8
Objective To investigate the microecological status of vaginal microflora in the women with different vaginal symptoms. Methods From March 2006 to October 2007, 6982 cases with varying degree vaginal symptoms including pruritus, increaseed leucorrhea, the leucorrhea having unusual smell, in the gynecology outpatient department were studied. The vagina secretions were examined in terms of the pH value, the hydrogen peroxide test, and Gram dyeing inspection of vaginal bacteria and microecology appraisal for colony's density, the multiplicity, the superiority fungus, and the inflammatory response. Results Among 6982 patients, normal vaginal microecology was identified in 750 (10. 74%, 750/6982 );abnormal mieroeeology was found in 6232 ( 89. 26%, 6232/6982 ) ; bacterial vaginosis (BV) was detected in 729 (10. 44%, 729/6982);vulvovaginal candidiasis (VVC) was in 1527 ( 21.87%, 1527/6982 ). Ninety five patients (1.36% ,95/6982) were with both BV and VVC. Abnormal bacteria colonies were found in 1229(17. 60% ,1229/6982),and others were found in 2652(37. 98% ,2652/6982). Conclusions The vaginal microecology in the women with different vaginal symptoms can be either normal or abnormal. Microecology clinical evaluation system can assess the vaginal microecosystem.
7.Expression of Notch1, Jagged1 and NICD in epithelial ovarian carcinomas and a preliminary study on the activity of gamma-secretase in epithelial ovarian carcinoma cell lines
Zhaoyi FENG ; Hongwu WEN ; Qinping LIAO
Chinese Journal of Obstetrics and Gynecology 2014;49(10):780-786
Objective To study the expression of Notch 1,Jagged1 and Notch intracellular domain (NICD) in epithelial ovarian carcinoma tissues and analyze the clinical significance.To explore the activity of γ-secretase in epithelial ovarian carcinoma cell line SKOV3 and the effect of N-[N-(3,5-dil uorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester (DAPT),a γ-secretase inhibitor on the activity of γ-secretase in SKOV3.Methods Immunohistochemistry staining method was performed in 43 patients with epithelial ovarian carcinoma and 11 patients with benign epithelial ovarian tumor to detect the expression of Notch1,Jagged1 and NICD.The differences of expressionof Notch1,Jagged1 and NICD between malignant and benign ovarian tumors was compared and alsoanalyzed the correlation with clinicopathological parameters of ovarian carcinoma.Human serous ovarian cancer cell line SKOV3 and immortalized nontumorigenic ovarian epithelial cell line T29 were incubated in vitro.The activities of γ-secretase in SKOV3 and T29 with dimethyl sulfoxide (DMSO) and DAPT were detected respectively by Gal4VP16/UAS and dual luciferase reporter assay system.Results (1) The immunohistochemical composite scores (ICS) of Notch1 in epithelial ovarian carcinoma (6.7±2.2) were not significantly different with those in benign epithelial ovarian tumor (5.4± 2.7,P=0.153),while the ICS of Jagged 1 and NICD in epithelial ovarian carcinoma (5.3± 2.4,5.3± 2.3) were higher than those in benign epithelial ovarian tumor (1.6± 1.4,3.1± 1.7; all P<0.01).The expression of Notch 1,Jagged 1 and N ICD had no correlation with patients' aged,history of carcinoma,ascites,the level of serum CA125,maximum length of ovarian tumor,Federation International of Gynecology and Obstetrics (FIGO) stage,grade and pathology subtypes (all P>0.05).The hazard ratio between the high expression of Notch1,Jagged1,or NICD and the moderate to low expression of Notch1,Jagged1,or NICD,and Jagged1 were 0.771,1.648 and 1.316,respectively (all P>0.05).The 5-year survival rate and median survival time between the high expression of Notch,Jagged 1 or NICD in subgroup and moderate to low expression in subgroup were of no difference (all P>0.05).The activity of γ-secretase in SKOV3 was significantly higher than that in T29 [(12.2± 1.4)%,P=0.019].(2)After DAPT treated,the relative activity of γ-secretase in SKOV3 (50 μmol/L) was declined from (100.0±5.3)% to (6.6±0.8)% (P=0.001).Conclusions Jagged1 and NICD in Notch1 pathway may play a key role in the occurrence of ovarian carcinoma.The activity of γ-secretase in epithelial ovarian carcinoma was higher than that in ovarian epithelial cell which suggest that DAPT,γ-secretase inhibitor,may become the target of ovarian carcinoma treatment.
8.Survey of sexual activity in pregnant women during the third trimester
Miao ZHANG ; Qinping LIAO ; Fengxia WANG
Chinese Journal of Perinatal Medicine 1998;0(02):-
Objective To investigate the sexual activity of pregnant women in the third trimester and its influencing factors. Methods A retrospective investigation was conducted among 338 postpartum women in our hospital who delivered between June and August in 2003. The questionnaires was completed by every subjects supervised by special trained persons on the 2nd to 4th day postpartum. Results More than a quarter (96/338, 28.4%) of the pregnant women had coitus during the third trimester. The increase of sexual activity was positively related to their partners who were less educated (Wald ?~2=9.68,P
9.Investigate the Pathogen Flora and the Treatment of Recurrent Vulvovaginal Candidiasis
Zhaohui LIU ; Xiaoli WANG ; Qinping LIAO
Journal of Practical Obstetrics and Gynecology 2009;25(12):730-732
Objective:To investigate the pathogen flora of recurrent vulvovaginal candidiasis(RWC),and to discuss its treatment. Methods: Fungal culture and susceptibility tests of vaginal discharge from 108 RWC patients were performed. Sensitive drugs were selected for effective consolidating treatment for half year. Fungal smears were examined at the 3rd and 6th months after the treatment respectively. Results:①Among RWC patients,there were 81 cas es of Candida albicans(75.0%), 19 cases of C. Glabrata(17.6%), 4 cases of C. Parapsilosis(3.7%) ,2 cases of C.tropicalis (1.9%) and 2 cases of M. Krusei (1.9%). The drug susceptibility test showed that 80.6% of these 108 patients were sensitive to nystatin; 66.7% were sensitive to clotrimazole; 65.7% were sensitive to miconazole; 57.4% were sensitive to fluconazole;8.3% were sensitive to ketoconazole;6.5% were sensitive to itraconazole; 7.4% were resistant to all of the above drugs.②The score of symptom and sign was significantly improved compared with the first visit at the 3rd and 6th month after the treatment( P<0.0001). The negative rate was 76.2%(80/105) at the 3rd month after the treatment and 64.8%(68/105)at the 6th month after the treatment. Conclusions:①The rate of non Candida albicans is increased to 25% in RWC patients, and 17.60% is Candida glabrata.②During the treatment of RWC, the drugs should be selected according to fungal culture, results of susceptibility test, medication experience of patients,and attack character. The research of immune abnormality of the local vagina will be helpful to improve the cure rate of RWC.
10.Effect of 17α-hydroxyprogesterone caproate and medroxyprogesterone acetate on inflammation-induced preterm birth of mouse model
Xiaoxiao ZHANG ; Chunyan SHI ; Qinping LIAO
Chinese Journal of Perinatal Medicine 2011;14(10):618-623
Objective To study the effect of 17α-hydroxyprogesterone caproate (17P) and medroxyprogesterone acetate (MPA) on expression of tumor necrosis factor-α (TNF-α)and cyclooxygenase-2 (COX-2) in placenta and uterine myometrium of inflammation-induced preterm birth mouse model to investigate the mechanism of preventing inflammation-induced preterm birth by progestogen.Methods Thirty clean CD-1 mice were divided into 6 groups (5 mice in each group) at 15th day of gestation:control group,lipopolysaccharides (LPS) group,17P 1 mg+LPS group,17P 2 mg+ LPS group,MPA 1 mg+LPS group and dimethyl sulfoxide (DMSO) + LPS group.Progestogens at different dosage were administered 1 h before LPS and 6 h after LPS administration.After these mice were sacrificed,TNF-α and COX-2 levels in the myometrium and placenta were detected by real-time PCR and immunohistochemistry.Data were analyzed by ANOVA,and comparisons between groups were adopted LSD method.Results 1.The comparison of relative expression of COX-2 mRNA and TNF-α mRNA in myometrium and placenta among groups:(1) Expressions of COX-2 mRNA and TNF-α mRNA in myometrium and placenta in the study groups were obviously higher than those of control group (P<0.05).(2) COX-2 mRNA expression in myometrium of 17P 1 mg+LPS group (11.410±3.931),17P 2 mg+LPS group (8.352±3.209) and MPA 1 mg+LPS group (11.920± 2.905) were obviously lower than that of LPS group (20.540± 4.147) and DMSO+ LPS group (18.620 ± 4.156) (P<0.05,respectively) ; although TNF-α mRNAexpression had similar trends among these groups,there were no statistical significance (P>0.05,respectively).(3) COX-2 mRNA expression in placenta of 17P 1 mg+LPS group (10.864±3.777),17P 2 mg+LPS group (7.084±1.667) and MPA 1 mg+LPS group (11.830±3.652) were obviously lower than that of LPS group (18.920±4.106) and DMSO+LPS group (23.820±7.554)(P<0.05,respectively).(4) TNF-α mRNA expression in placenta in 17P 1 mg+LPS group (14.340±1.618),17P 2 mg+ LPS group (11.488 ± 2.910) and MPA 1 mg+ LPS group (13.040 ± 2.982) were obviously lower than that of LPS group (24.240±7.059) and DMSO+LPS group (23.040±5.896) (P<0.05,respectively).2.The comparison of protein expression of COX-2 and TNF-α in placenta among groups:(1) The expression of COX-2 protein in placenta of the study groups was significantly higher than that of control group (P<0.05).(2) There were no differences among the COX-2 protein expression of 17P 1 mg+ LPS group (14 360.92± 1766.01),17P 2 mg+ LPS group (13 340.18±965.35) and MPA 1 mg+LPS group (12 870.81±1521.97)(P>0.05),while the COX-2 protein expressions of them were significantly lower than that of LPS group (16 426.64 ± 1823.87) and DMSO+LPS group (16 761.23±2388.17)(P<0.05,respectively).(3) There were no differences among the TNF-α protein expression of 17P 1 mg+LPS group (22 750.96±4656.68),17P 2 mg+LPS group (22766.24± 3500.34) and MPA 1 mg+LPS group (20770.01±3318.48)(P>0.05),while the TNF-α protein expressions of them were significantly lower than that of LPS group (26204.49±5090.34) and DMSO+LPS group (27346.18±3269.30)(P<0.05,respectively).Conclusions 17P and MPA might prevent the preterm parturition of inflammation-induced mouse model by inhibiting inflammation cytokines and prostaglandins.