Objective: To investigate the differences in enolase and laminin levels in vaginal epithelial tissues between mice successfully infected withGardnerellaand mice not infected with Gardnerella, providing information for further exploration of the correlation between enolase and laminin levels and the incidence of bacterial vaginosis. Methods: Gardnerella strains isolated, purified, and identified from vaginal secretions of patients with bacterial vaginosis were used to infect the vagina of mice and establish a mouse model of bacterial vaginosis. Successful and failed mice was defined as successful and failed groups, respectively. Differential expression of enolase and laminin in the vaginal epithelial tissue of two groups of mice was detected by Western blot. Modeling success rate was statistically analyzed, and the expression differences of enolase and laminin was compared between two groups. Results: One strain of Gardnerella vaginalis infected 10 SPF grade KM mice, 7 mice met the diagnostic criteria for bacterial vaginosis, and 3 mice failed to model, with a success rate of 70%. Western blot was used to detect protein expression levels, and the levels of laminin and enolase in the successfully modeled mouse vaginal epithelial tissue were significantly higher than those in the failed modeling group, with statistical differences between the two groups(P<0.05). Conclusion Enolase and laminin may be involved in the occurrence of bacterial vaginosis, however, further research is needed to determine the mechanisms through which they trigger the occurrence and development of the disease.

Objective:Using clotrimazole vaginal tablet as a positive control, to evaluate the results of clotrimazole vaginal expansion suppository in the treatment of mild and moderate vulvovaginal candidiasis in terms of efficacy, patient satisfaction, side effects, and recurrence rate.Methods:This study was jointly conducted by 5 hospitals from August 2017 to October 2018, patients with mild and moderate vulvovaginal candidiasis confirmed by fungal culture and symptoms scores were selected. They were randomized to experimental group and control group as 1∶1 ratio. In the experimental group ( n=105), the subjects applied clotrimazole vaginal expansion suppository (150 mg) daily at night for 7 days. In the control group ( n=106), the subjects used a single dose of clotrimazole vaginal tablet (500 mg). Follow-ups were performed at (8±3) and (30±5) days after the discontinuation of the drugs, respectively. The difference in clinical symptoms and signs scores was used to evaluate the improvement of clinical symptoms, and the patient′s satisfaction and side effects were recorded. Results:At the first follow-up, the experimental group and control group were followed up by fungal culture on the cure rate [66.7% (70/105) versus 63.2% (67/106), P>0.05] and total effective rate [98.1% (103/105) versus 99.1% (105/106), P>0.05], the differences were not statistically significant. At the second follow-up, the recurrence rates of the experimental group and the control group were 5.7% (4/70) and 14.9% (10/67), respectively, with no significant difference ( P>0.05). In the evaluation of patient satisfaction, the leakage of the drug in the experimental group was significantly better than that in the control group ( P<0.01). The side effects mainly included vaginal stimulation, itching and burning sensation, and there was no statistical difference between the two groups ( χ2=1.070, P=0.586). Conclusions:In the treatment of mild and moderate vulvovaginal candidiasis, clotrimazole vaginal expansion suppository is no less effective than clotrimazole vaginal tablet, and there is no significant difference in the recurrence rate between the two. In terms of patient satisfaction, clotrimazole vaginal expansion suppository is superior to clotrimazole vaginal tablet.

Reproductive tract infection is an important part of systemic infection,and it has become a major social and public health problem in the world.Recent studies have shown that reproductive tract infection is closely related to tumor occurrence, infertility, ectopic pregnancy, premature rupture of membranes and premature delivery and so on.Almost all reproductive tract infections are accompanied by changes of genital tract microenvironment.The development of the vaginal microecological detection system and the application of the diagnosis platform for the lower genital tract make it possible toaccurately treat female genital tract infection.Therefore, more and more attention was also paid to the standardized test of female reproductive tract infection and vaginal microecology by experts and scholars in the related fields. (Chin J Lab Med,2018,41:251-253)

Objective To study the vaginal microecology of the patients in the outpatient department of Obstetrics and gynecology in China.Methods A multicenter cross-sectional study was conducted in gynecologic clinic of 9 collaborative hospitals in China.200 consecutive patients were collected in each hospital and their vaginal microecology combined with related factors were analyzed.Results A total of 2 093 specimens were collected in this study.The detection rate of Trichomonas was 5.5%(115/2 093). The detection rate of Candida mycelia was 15.9%(333/2 093), with germinal spores was 4.1%(86/2093).The detection rate of bacterial vaginosis was 18.8%(394/2 093).The distribution results of vaginal flora in patients showed that the normal flora accounted for only 27.3%(571/2 093).The normal flora with the insufficiency of H2O2 accounted for 23%(480/2 093).The bacteria inhibiting flora accounted for 3.8%(79/2 093).The abnormal microflora(non BV type)accounted for 14.9%(312/2 093).The abnormal microflora(BV intermediate type)accounted for 13.4%(280/2 093).The abnormal microflora(BV type)accounted for 17.6%(369/2 093).The average pH of vaginal discharge was 4.58 ±0.495.There was no significant difference of the incidence of trichomonas and bacterial vaginosis between north and south of the Yangtze river, while the detection rate of fungal hyphae and the fungal spores is significantly higher in the south than that in the north.The analysis results of factors affecting the microecology showed that age and contraception methods were two important factors.The patients′age from bacteria inhibition group was 49.64 +16.68 which was significantly higher than that of the other microecology groups.The proportion of abnormal microflora of patients from the oral contraceptive group was 40%(20/50).The proportion of abnormal microflora of patients from IUD group was 36.6%(63/172).Compared with these two contraception methods, the proportion of abnormal microflora of patients from condoms usage group was 27.8%(91/327)which was significantly lower.The incidence of abnormal leucorrhea in the normal group was 37.7%, which was significantly lower than that of other abnormal groups.Conclusion This study showed the vaginal microecology status of the Chinese outpatient ′s clinic and found that the vagina microecology was related to age, region and contraceptive methods.The typical manifestation of microecological abnormality is the increase of leucorrhea.(Chin J Lab Med,2018, 41:287-291)

Objective To evaluate the efficacy,bleeding profile and safety of low-dose levonorgestrel-releasing intrauterine system (LNG-IUS 8) in Chinese healthy women of childbearing age.Methods A multi-center,open-label,single-arm clinical trial conducted at 16 centres in China enrolled 773 healthy women of childbearing age (mean age 31.6 years old,range 18 to 40 years old),who demanded contraception,from April 2006 to June 2013.All women placed LNG-IUS 8 for 3 years and then been followed up at 3,6,9,12,18,24,30,36 months.The efficacy variables including pregnancy rate and expulsion rate were analyzed using life table,while observing adverse events (AE) to evaluate the safety.The bleeding profile happened during the study was assessed using 90-day reference intervals (World Health Organization criteria).Results Eight pregnancies occurred among 773 women,resulting in a overall Pearl index of 0.42 per 100 women years.The 3-year cumulative pregnancy rate was 0.37 per 100 women years and the 3-year cumulative expulsion rate was 1.99 per 100 women years.The number of women with bleeding/spotting reduced and the bleeding/spotting days declined over time.Totally 219 AE were reported related to LNG-IUS 8 placements.The most common AE were vaginal bleeding (8.2％,63/773)and the ovarian cyst (6.2％,52/773).LNG-IUS 8 had an improving effect on dysmenorrhea that the percentage of women with dysmenorrhea as well as the days of dysmenorrhea decreased over time.The percentage of women satisfied or very satisfied with LNG-IUS 8 was 87.2％ (622/713).Conclusion LNG-IUS 8 is highly effective and safe for Chinese healthy women of childbearing age.

Objective To investigate the variants presence and intratype lineages distribution of Human papillomavirus type 52 (HPV52) in Chinese women.Methods A total of 1 244 cervical specimens were collected.After DNA extraction and genotyping,79 HPV52 single positive DNA specimens were analyzed for E7 oncogenes and Long Control Region (LCR) fragments sequence variations.Finally,50 samples were sequenced for both of the two fragments.Constituent ratio of HPV52 Lineage distribution was compared between mainland China and other regions of Asia.Results DNA variation of E7 and LCR were detected in 92.0％ (46/50) and 96.0％ (48/50) of the cases respectively.Five and fourteen variants of HPV52 E7 and LCR were found respectively.Lineage B was the most prevalent lineage across all of the HPV52 single infection cases (90.0％,45/50),lineage A accounted for 8.0％ (4/50),and lineage C accounted for 2.0％ (1/50),and no lineage D was found.Constituent ratio of HPV52 Lineage distribution was different in different regions of Asia.Conclusions There is a high frequency of mutation in HPV52 E7 and LCR in China.Constituent ratio of HPV52 Lineage distribution of Asia has the character of geographical dependence.

Objective:To investigate the female vaginal Lactobacillus crispatus biofilm by using confocal laser scanning microscopy (CLSM),thus revealing the formation of biofilm. Methods:The cover slide biofilm culture approach in vitro was employed for induction of the vaginal Lactobacillus crispatus biofilm formation. Following the culture for 2, 4, 8, 12, 16, 20, 24, 48, 72, 96 and 120 hours, the cover slide was removed for subsequent staining with the fluoresce in isothiocyanate-conjugated concanavalin A ( FITC-ConA) and propidium ( PI) . This was followed by determination of the formation and characteris-tics of the vaginal Lactobacillus crispatus biofilm by using CLSM. Results:The CLSM images of biofilm formation at different time points were captured, suggesting that the vaginal Lactobacillus crispatus adhe-sion occurred at h 4, which was in reversible attachment, then more and more Lactobacillus crispatus ag-gregated at h 8 to h 20, which was in irreversible attachment. Lactobacillus crispatus clustered at h 20, with early development of biofilm architecture. Then the biofilm with extracellular matrix around the bacte-ria was set up at h 24,with gradual matureation at h 24 to h 48. The biofilm dispersed at h 72. The bio-film density of cultivating for 20 hours was 42 . 7 × 10 -3 ± 6 . 8 × 10 -3 , and for 24 hours increased to 102. 5 × 10 -3 ± 23. 1 × 10 -3 , suggesting a significant difference, P<0. 05. This meant that mature bio-film was formed at h 24 . Conclusion:The vaginal Lactobacillus crispatus is able to form typical biofilm with distinct developmental phases and architecture characteristics. Mature biofilm is formed at h 24 to h 48, then the biofilm begins to disperse.

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.

Objective To investigate the effects of postoperative adjuvant chemotherapy (CT) and chemoradiotherapy (CRT) or radiotherapy(RT) for Ⅰ b-Ⅱ a cervical cancer with risk factors.Methods From March 1995 to June 2010,there were 137 patients underwent radical hysterectomy and systematic pelvic lymphadenectomy for stage Ⅰ b-Ⅱ a cervical cancer admitted at Peking University First Hospital.These patients had risk factors,intermediate risk factors including bulky tumor (＞4 cm),lymph vascular space invasion,deep stromal invasion; high risk factors including positive surgical margin,parametrial invasion,lymph node involvement.Of the all patients,79 cases of them were treated with CT,58 of them were treated with RT or CRT.The 5-year survival and prognosis factors were analyzed retrospectively,the prognosis was compared between two adjuvant therapy groups.Results The univariate analysis shown that types of pathology,different grade of risk factors,stroma invasion and lymph node involvement were prognostic factors of 5-year overall survival Patients with squamous cell carcinoma,intermediate risk factors,no parametrial invasion,and no lymph node involvement had better prognosis (P ＜ 0.05).Whether patients with high-risk factors or intermediate-risk factors,the 5-year overall survival and 3-year disease-free survival had no difference between CT and RCT or RT groups respectively.Cox regression multivariate analysis of survival indicated that clinical stages,types of histology,different grade of risk factors were independent prognostic indicator.Patients with early stage,squamous cell carcinoma,intermediate risk factors had better prognosis.Univariate and multivariate analysis indicated that different postoperative adjuvant therapies had no effects on the prognosis.The 5-year overall survival was 88.6％ in patients treated with CT,and 89.7％ in patients treated with RT or CRT (P =0.455).Conclusion There are equivalent therapeutic results between CT and RT or CRT for patients with risk factors after radical surgery,CT may be as one choice of postoperative adjuvant therapy for stage Ⅰ b-Ⅱ a cervical carcinoma with risk factors.

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.