[Analysis of homology and drug sensitivity of vaginal isolates of 10 patients with recurrent vulvovaginal candidiasis in recurrent episodes].

Objective: To detect karyotype homology of vaginal isolates from patients with recurrent vulvovaginal candidiasis (RVVC) in recurrent episodes, and to discuss changes of susceptibility of Candida strains to antifungal drugs with clinical progress. Method: s Ten patients were recruited from Beijing Obstetrics and Gynecology Hospital, Capital Medical University from September 2018 to June 2019, who were firstly diagnosed with RVVC. Vaginal discharges were collected before first treatment and after first relapse. Vaginal strains were isolated, purificated and identificated. Then karyotype of 20 strains isolated from 10 patients were detected by restriction endonuclease analysis of genomic DNA (REAG) using enzyme BssHⅡand pulsed field gel electrophoresis (PFGE) methods, and sensitivity of clinical isolates to 5 antifungal drugs (clostridium, fluconazole, miconazole, itraconazole and nystatin) was also detected using disk diffusion method. Result: s (1) All 20 strains of 10 patients with RVVC were Candida albicans, and their chromosomes were extremely similar after BssHⅡ enzyme digestion. The gene bands of isolated strains from the same patient were completely identical. (2) After clinical medication, the sensitivity of vaginal isolates to azoles was generally decreased, but remained highly sensitive to nystatin, nystatin (first and second clinical isolates: 100% sensitivity and 100% sensitivity)>clotrimazole (100% sensitivity and 90% sensitivity)>fluconazole (80% sensitivity and 70% sensitivity)>itraconazole (60% sensitivity and 50% sensitivity)>miconazole (30% sensitivity and 20% sensitivity). Conclusions: (1) The latency of the same colonized strain in the vagina may be the cause of repeated RVVC episodes. (2) Antifungal agents could selectively induce drug resistance to Candidas, and Candidas show cross-resistance to antifungal agents. Repeated fungal culture and drug sensitivity test in patients with RVVC are very necessary for correct selection of antifungals.

[Changes of local vaginal immune regulation in rats infected with vulvovaginal candidiasis].

Objective: To study the disease process of vulvovaginal candidiasis (VVC) infection in rat model of VVC, and to study the immuno-repairing effect of different treatments on vaginal epithelium and the ultra-structural changes of vaginal epithelial cells. Methods: The VVC model of female rats were established. After successful modeling, the rats were treated with no treatment (model control group), nystatin and Kangfu Xiaoyan suppository. The vaginal epithelium was observed by transmission electron microscopy and immunohistochemical staining. The ultra-structural changes of epithelial cells and the expression of cytokines interferon γ (IFN-γ), interleukin (IL) 4, IL-17 and IgG in epithelial cells were observed and analyzed statistically. Results: The negative conversion rate of model control group was 0, and that of nystatin group was 6/6, and that of Kangfu Xiaoyan suppository group was 5/6; significant difference existed between nystatin, Kangfu Xiaoyan suppository group and model control group (P<0.05). The ultrastructures of vaginal epithelial cells were damaged obviously after VVC infection, and the ultrastructures were repaired by nystatin and Kangfu Xiaoyan suppository under transmission electron microscope. Immunohistochemical staining showed, the expressions of IFN-γ and IgG in the four cytokines which played a protective role increased after Kangfu Xiaoyan suppository treatment, significantly different from that of model control group (P<0.05), but there were no significant differences of the IFN-γ and IgG expression between Kangfu Xiaoyan suppository group and those of nystatin group (P>0.05); the expression of IL-17 was increased after nystatin treatment, but decreased after Kangfu Xiaoyan suppository treatment, and the difference between the two groups had statistical significance (P<0.05). Conclusions: The ultrastructure of vaginal epithelial cells after VVC infection could be damaged obviously, the local immune state is disordered, and the antifungal drug nystatin has a good therapeutic effect on VVC, it could significantly repair the damaged vaginal epithelium structure after VVC infection and strengthen the protective immune function of vaginal epithelium. Kangfu Xiaoyan suppository, one of Chinese medicine, has similar therapeutic effect with nystatin.

[Effects of metformin and adiponectin on endometrial cancer cells growth].

OBJECTIVE: To determine the effect of metformin and adiponectin on the proliferation of EC cells and the relationship between metformin and adiponectin. METHODS: The proliferation impact of different concentrations of metformin and adiponectin on two types of EC cells ishikawa (IK) and HEC-1B was confirmed by CCK-8 method. qRT-PCR and Western blot were used to detect the effect of different concentrations of metformin on the changes of adiponectin receptors (AdipoR1 and AdipoR2) of the EC cells both in mRNA and protein level and the role of compound C, an adenosine monophosphate-activated protein kinase (AMPK) inhibitor, on the above effects. RESULTS: (1) Both metformin and adiponectin could significantly promote the proliferation of endometrial cancer (EC) cells in a time and concentration dependent manner (P<0.05).(2)Metformin and adiponectin had synergy anti-proliferative effect on EC cells and the combination index (CI) value of IK cells was 0.906 34 and of HEC-1B cells was 0.827 65. (3)qRT-PCR was used to detect the mRNA levels of AdipoR1 and AdipoR2 after 5 mmol/L and 10 mmol/L metformin, respectively, stimulating IK and HEC-1B cells for 48 hours and the mRNA expressions of AdipoR1 and AdipoR2 were significantly increased when compared with the control group (0 mmol/L)(IK: AdipoR1 of 5 mmol/L and 10 mmol/L group: P<0.001,AdipoR2 of 5 mmol/L group: P<0.001; HEC-1B: AdipoR1 of 5 mmol/L group: P<0.001, 10 mmol/L group: P=0.023, AdipoR2 of 5 mmol/L group: P<0.001, 10 mmol/L group: P=0.024). When combined with compound C, the RNA levels of AdipoR1 and AdipoR2 were not different compared with the control group (0 mmol/L, P>0.05). (4) Western blot was used to detect the protein levels of AdipoR1 and AdipoR2 after 5 mmol/L and 10 mmol/L metformin, stimulating IK and HEC-1B cells for 48 hours and the protein level was significantly increased when compared with the control group (0 mmol/L)(IK: AdipoR1 of 5 mmol/L group: P=0.04, 10 mmol/L group: P=0.033, AdipoR2 of 5 mmol/L group: P=0.044, 10 mmol/L group: P=0.046; HEC-1B: AdipoR1 of 5 mmol/L group: P=0.04, 10 mmol/L group: P=0.049, AdipoR2 of 5 mmol/L group: P=0.043, 10 mmol/L group: P=0.035). When combined with compound C,the protein levels of AdipoR1 and AdipoR2 were not different compared with the control group (0 mmol/L, P>0.05). CONCLUSION: We find that metformin and adiponectin have synergy anti-proliferative effect on EC cells. Besides, metformin can increase adiponectin receptors expressions of EC cells both in mRNA and protein levels and this effect is accomplished by the activation of AMPK signaling pathway.

Molecular analysis of the relationship between specific vaginal bacteria and bacterial vaginosis metronidazole therapy failure.

Bacterial vaginosis frequently persists, even after treatment. The role of some strains of bacteria associated with bacterial vaginosis treatment failure remains poorly defined. The aim of our study was to define the risk of bacterial vaginosis treatment failure, including pre-treatment detection of specific vaginal bacteria. Bacterial vaginosis is present when the Nugent score is ≥7 and the modified Amsel criteria is positive. Women with bacterial vaginosis were treated with intravaginal metronidazole gel nightly for 5 nights. The 454 pyrosequencing method was used to detect bacteria in vaginal fluid. By univariate analysis, a history of bacterial vaginosis, intrauterine device use and the presence of Facklamia, Corynebacterium and Veillonella were significantly associated with bacterial vaginosis treatment failure. Lactobacillus crispatus, Lactobacillus pentosus and Megasphaera were significantly associated with curing bacterial vaginosis. After logistic regression analysis and detection of these bacteria for test-of-cure, we found that women who had a history of bacterial vaginosis had a higher incidence of bacterial vaginosis treatment failure, whereas women with L. crispatus had a lower incidence of treatment failure. Post-treatment sexual activity was not associated with the treatment effect. Our data suggested that treatment failure may be not caused by drug resistance. Rather, it has a closer relationship with the failed restoration of lactobacilli.