Objective To investigate the vaginal flora in patients with recurrent vulvovaginal candidiasis (RVVC).Methods Vaginal swabs were collected at different time points from 6 RVVC patients and 5 healthy women of child-bearing age.The dynamic changes,microbiota composition,alpha diversity and beta diversity in the two groups were assessed by analyzing the 16S rRNA V4 hypervariable region amplified from the total genomic DNA from the swabs.Results Lactobacillus was the predominant species in healthy women with similar proportions of Liners and L.crispatus;small proportions of Gardnerella,Prevotella and other genus were also detected.In some healthy women,the vaginal flora showed a high relative abundance of anaerobic bacteria such as Gardnerella,Prevotella,Atopobium,Sneathia.Compared with the healthy women,patients with RVVC showed a significantly reduced diversity of vaginal flora,where Liners was the predominant species and the content of L.crispatus decreased significantly.In healthy women,the vaginal flora fluctuated with the menstrual cycle,and the fluctuation was the most prominent during menstruation;the dominant species either alternated regularly or maintain an absolute superiority in the menstrual cycle.The vaginal flora showed attenuated fluctuation in women with RVVC,were highly conserved within the menstrual cycle,and maintained a similar composition in the episodes and intermittent periods.Conclusion The vaginal flora of RVVC patients do not undergo regular variations with the menstrual cycle and shows a similar composition between the episodes and intermittent periods.Promoting the production of L.iners or inhibiting the colonization of L.crispatus to restore the composition of the vaginal flora may help in the treatment of RVVC.

Objective To investigate the vaginal flora in patients with recurrent vulvovaginal candidiasis (RVVC).Methods Vaginal swabs were collected at different time points from 6 RVVC patients and 5 healthy women of child-bearing age.The dynamic changes,microbiota composition,alpha diversity and beta diversity in the two groups were assessed by analyzing the 16S rRNA V4 hypervariable region amplified from the total genomic DNA from the swabs.Results Lactobacillus was the predominant species in healthy women with similar proportions of Liners and L.crispatus;small proportions of Gardnerella,Prevotella and other genus were also detected.In some healthy women,the vaginal flora showed a high relative abundance of anaerobic bacteria such as Gardnerella,Prevotella,Atopobium,Sneathia.Compared with the healthy women,patients with RVVC showed a significantly reduced diversity of vaginal flora,where Liners was the predominant species and the content of L.crispatus decreased significantly.In healthy women,the vaginal flora fluctuated with the menstrual cycle,and the fluctuation was the most prominent during menstruation;the dominant species either alternated regularly or maintain an absolute superiority in the menstrual cycle.The vaginal flora showed attenuated fluctuation in women with RVVC,were highly conserved within the menstrual cycle,and maintained a similar composition in the episodes and intermittent periods.Conclusion The vaginal flora of RVVC patients do not undergo regular variations with the menstrual cycle and shows a similar composition between the episodes and intermittent periods.Promoting the production of L.iners or inhibiting the colonization of L.crispatus to restore the composition of the vaginal flora may help in the treatment of RVVC.

Objective To observe the therapeutic effect of NF-κB gene short hairpin RNA (shRNA) on endometriosis and identify the function of NF-κB on the maintenance and development of endometriosis in Macaca fascicularis.Methods The Macaca fascicularis model of endometriosis was developed,which divided into experimental group,negative control group and simple model group.The high specificity adenovirus vector mediated shRNA targeting NF-κB gene and negative control shRNA adenovirus with no-load NF-κB gene were synthesised.The experimental group injected the adenovirus which carried the NF-κB shRNA into the endometriosis lesions under laparoscopy surgery,the negative control group with no-load shRNA adenovirus and the simple models group injected with normal saline.Four weeks later after the injection,an observed operation was performed through laparoscopy and some lesions were collected.The CD34 immunohistochemistry of these lesions were done to detect the microvessel density,then the variation of the microvessel density among each group were observed.The expression of the NF-κB and proliferating cell nuclear antigen (PCNA) were detected through western blot.Results First,the Macaca fascicularis model of endometriosis was successful developed,and the experimental group has an evident atrophy in ectopic lesions compared with the previous.The lesions' microvessel density in experimental group decreased evidently compared with the negative control group and simple model group (0.002 0±0.000 3 versus 0.021 9±0.002 6 versus 0.024 5±0.003 3),and the differences was statistically significant (P＜0.01).The expression of PCNA (0.37±0.17 versus 0.57±0.26 versus 0.57±0.28) and NF-κB (0.338 ± 0.174 versus 0.678 ± 0.021 versus 0.645 ±0.098) in experiment group was lower than the negative control group and simple model group,the differences were statistically significant (all P＜0.01).Conclusion Through targeting suppressed the NF-κB gene expression by NF-κB shRNA,we can inhibit the development of endometriosis through reducing the ability of angiogenesis and cell proliferation of ectopic endometrial cells.

OBJECTIVETo assess the effect of a high specific adenovirus vector-mediated shRNA targeting nuclear factor-κB (NF-κB) on cell proliferation of the endometrium of Macaca fascicularis.

METHODSThe adenoviral vector NF-κB-p65-shRNA and the empty vector were separately trasnfected in cultured endometrial cells of Macaca fascicularis. The changes in the expression of the target gene protein and apoptotic proteins, cell proliferation, and cell cycle distribution were observed after the transfection.

RESULTSCompared with the control cells, infection of the endometrial cells with the NF-κB-p65-shRNA adenovirus significantly increased the expression levels of apoptotic proteins, promoted apoptosis of the endometrial cells, and reduced the cells in division?stage.

CONCLUSIONSNF-κB-p65 shRNA adenovirus can effectively promote apoptosis of endometrial cells and inhibit the proliferation of endometrial cells of Macaca fascicularis.

Adenoviridae ; Animals ; Apoptosis ; Cell Proliferation ; Cells, Cultured ; Endometrium ; cytology ; Female ; Genetic Vectors ; Macaca fascicularis ; RNA, Small Interfering ; genetics ; Transcription Factor RelA ; genetics ; Transfection

Objective To assess the effect of a high specific adenovirus vector-mediated shRNA targeting nuclear factor-κB (NF-κB) on cell proliferation of the endometrium of Macaca fascicularis. Methods The adenoviral vector NF-κB-p65-shRNA and the empty vector were separately trasnfected in cultured endometrial cells of Macaca fascicularis. The changes in the expression of the target gene protein and apoptotic proteins, cell proliferation, and cell cycle distribution were observed after the transfection. Results Compared with the control cells, infection of the endometrial cells with the NF-κB-p65-shRNA adenovirus significantly increased the expression levels of apoptotic proteins, promoted apoptosis of the endometrial cells, and reduced the cells in division stage. Conclusions NF-κB-p65 shRNA adenovirus can effectively promote apoptosis of endometrial cells and inhibit the proliferation of endometrial cells of Macaca fascicularis.

Objective To assess the effect of a high specific adenovirus vector-mediated shRNA targeting nuclear factor-κB (NF-κB) on cell proliferation of the endometrium of Macaca fascicularis. Methods The adenoviral vector NF-κB-p65-shRNA and the empty vector were separately trasnfected in cultured endometrial cells of Macaca fascicularis. The changes in the expression of the target gene protein and apoptotic proteins, cell proliferation, and cell cycle distribution were observed after the transfection. Results Compared with the control cells, infection of the endometrial cells with the NF-κB-p65-shRNA adenovirus significantly increased the expression levels of apoptotic proteins, promoted apoptosis of the endometrial cells, and reduced the cells in division stage. Conclusions NF-κB-p65 shRNA adenovirus can effectively promote apoptosis of endometrial cells and inhibit the proliferation of endometrial cells of Macaca fascicularis.

OBJECTIVETo study the effect of interleukin-1β (IL-1β) on the expressions activin A, follistatin, and cripto in cultured human endometrial stromal cells (HESCs) form patients with endometriosis.

METHODSCultured HESCs were stimulated with 250, 500, and 750pg/ml IL-1β, and the mRNA and protein expressions of activin A, follistatin, and cripto were assayed using real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay.

RESULTSIL-1β treatment caused significant dose-dependent increments of the mRNA and protein expressions of activin A and follistatin and of the mRNA expression of cripto in cultured HESCs.

CONCLUSIONIL-1β can affect the expressions of activin A, follistatin and cripto in HESCs from patients with endometriosis.

Activins ; metabolism ; Cells, Cultured ; Endometriosis ; metabolism ; Endometrium ; cytology ; Female ; Humans ; Interleukin-1beta ; pharmacology ; Stromal Cells ; drug effects ; metabolism

Objective To study the effect of interleukin-1β(IL-1β) on the expressions activin A, follistatin, and cripto in cultured human endometrial stromal cells (HESCs) form patients with endometriosis. Methods Cultured HESCs were stimulated with 250, 500, and 750 pg/ml IL-1β, and the mRNA and protein expressions of activin A, follistatin, and cripto were assayed using real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay. Results IL-1βtreatment caused significant dose-dependent increments of the mRNA and protein expressions of activin A and follistatin and of the mRNA expression of cripto in cultured HESCs. Conclusion IL-1βcan affect the expressions of activin A, follistatin and cripto in HESCs from patients with endometriosis.

Objective To study the effect of interleukin-1β(IL-1β) on the expressions activin A, follistatin, and cripto in cultured human endometrial stromal cells (HESCs) form patients with endometriosis. Methods Cultured HESCs were stimulated with 250, 500, and 750 pg/ml IL-1β, and the mRNA and protein expressions of activin A, follistatin, and cripto were assayed using real-time reverse transcriptase-polymerase chain reaction and enzyme-linked immunosorbent assay. Results IL-1βtreatment caused significant dose-dependent increments of the mRNA and protein expressions of activin A and follistatin and of the mRNA expression of cripto in cultured HESCs. Conclusion IL-1βcan affect the expressions of activin A, follistatin and cripto in HESCs from patients with endometriosis.

BACKGROUND: Lentivirus can infect divided and undivided cells. It remains uncertain whether the lentivirus can successful y infect primary ovarian granulosa cells. OBJECTIVE: To investigate infecting ratio and cel apoptosis of lentivirus carrying bcl-2 gene in primary human ovarian granulose cells cultured in vitro. METHODS: The lentiviral vector carrying bcl-2 gene was constructed using molecular biology, and packaged into lentivirus with high titer. The resulting recombinant lentivirus carrying bcl-2 genes were then used to infect primary human ovarian granulosa cells in vitro at different multiplicity of infection, 10, 50, 100, 200, and 400. Infection efficiency and cel proliferation were observed at 24, 48, 72, and 96 hours fol owing infection. Cel apoptosis was detected by flow cytometry, and bcl-2 gene transcription was assessed using reverse transcription PCR. RESULTS AND CONCLUSION: Primary human ovarian granulosa cells adhered at 24 hours, and exhibited polygon- or fusiform-shape and colony-like growth. When multiplicity of infection was 100, cel appearance and growth remained unchanged, and infection efficiency was high, which reached the peak up to 72 hours. Moreover, the positive rate was up to 60% in granulosa cells. Lentivirus carrying bcl-2 gene could increase expression of Bcl-2 protein and inhibit apoptosis of primary ovarian granulosa cells.