高危型HPV感染与阴道微生物组关系

目的 通过分析高危型人乳头瘤病毒（HPV）感染患者的阴道菌群,探讨HPV感染与阴道微生物组的关系。方法 选取于国际和平妇幼保健院体检中心体检的63例HPV阴性女性（对照组）及39例高危型HPV感染者（HPV感染组）为研究对象。对细菌16S rDNA的V1V2域扩增后,使用高通量测序法分析两组对象阴道微生物组的组成与结构差异。结果 在门水平上,两组对象均以厚壁菌门（Firmicutes）为主要优势门,其在感染组中占55.0%,在对照组中占88.4%。Alpha多样性指数分析表明感染组患者的阴道菌群多样性均高于对照组,差异有统计学意义（P<0.05）。LEfSe结果显示加德纳菌属在感染组低级别鳞状上皮内病变（LSIL）型患者中为优势菌,乳杆菌属为对照组优势菌。结论 高危型HPV感染者阴道菌群多样性增加。加德纳菌属为LSIL与HPV阴性女性的差异菌,乳杆菌属为HPV阴性女性阴道特征菌。     

Longitudinal analysis of the vaginal microflora in pregnancy suggests that L. crispatus promotes the stability of the normal vaginal microflora and that L. gasseri and/or L. iners are more conducive to the occurrence of abnormal vaginal microflora

Background  

Despite their antimicrobial potential, vaginal lactobacilli often fail to retain dominance, resulting in overgrowth of the vagina by other bacteria, as observed with bacterial vaginosis. It remains elusive however to what extent interindividual differences in vaginal Lactobacillus community composition determine the stability of this microflora. In a prospective cohort of pregnant women we studied the stability of the normal vaginal microflora (assessed on Gram stain) as a function of the presence of the vaginal Lactobacillus index species (determined through culture and molecular analysis with tRFLP).     

Spatial relationship of the vaginal microflora to the vaginal epithelium of female rats: scanning electron microscopy.

The vaginal mucosa of ovariectomized female rats has been examined by scanning electron microscopy before and after estrogen [1,3,5(10)-estratriene-3,17beta-diol] treatment. Without estrogen stimulation vaginal colonization is minimal and the epithelium is characterized by a layer of epithelial cells covered with small microvillous-like projections. Progressive changes that were consistent with estrogenic cytoproliferative effects were seen after estrogen treatment. By post-treatment day 3 bacterial colonization was maximal and the epithelium was comprised of flat squamous cells that tended to become detached from the underlying tissue layers. Bacteria were seen in association with the intercellular borders of this tissue and occurred singly or as microcolonies. No distinct physical attachment structures were identified, although an amorphous extracellular material that may serve to attach the bacteria to the squamous epithelial cells was frequently seen.     

门诊阴道镜检查患者阴道菌群构成与HPV16感染的关系

目的 探讨门诊阴道镜检查患者阴道菌群构成与HPV16感染的关系。 方法 选取我院2016年3月-2018年12月门诊阴道镜检查的高危HPV感染患者96例为研究对象,通过阴道镜检查结果分析其HPV16感染情况、阴道菌群构成及菌群多样性。 结果 HPV16阳性组人数41例,阴性组55例,感染率为42.7%。HPV16阳性组患者阴道菌群中占前5位的是乳酸杆菌属、加德纳菌属、嗜胨菌属、纤毛菌属及普氏菌属,阴性组占前5位的分别为乳酸杆菌属、加德纳菌属、奇异菌属、嗜胨菌属及卟啉单胞菌属。HPV16阳性组乳酸杆菌属构成比显著小于HPV阴性组,差异具有统计学意义(P=0.007,χ2=7.195),两组加德纳菌属、嗜胨菌属构成比无差异(P=0.073,χ2=3.217;P=0.763,χ2=0.091)。HPV16阳性组菌群Shannon指数、ACE指数、Chaol指数显著大于阴性组,差异具有统计学意义(P=0.014,χ2=2.461;P=0.002,χ2=3.025;P2=4.534);Simpson指数比较差异无统计学意义(P=0.310,χ2=0.406)。 结论 阴道菌群结构的改变主要表现为乳酸杆菌属数量的减少及加德纳菌属、嗜胨菌属、普氏菌属及纤毛菌属的增加,这可能与HPV16感染有一定关系。     

The characteristics of the vaginal microflora in intrauterine contraception

The results of the study of vaginal microbiocenosis in women using intrauterine contraceptives are presented. Inflammatory complications in intrauterine contraception were shown to be linked with vaginal dysbiosis. Disturbances in vaginal microbiocenosis were characterized by the deficiency of lactoflora and the presence of enterobacteria, Staphylococcus aureus, Gardnerella of fungi of the genus Candida. The problem of the possibility of complications of microbial etiology in women using intrauterine contraceptives are discussed.     

Intestinal microflora of the newborn rat as related to mammary, faecal, and vaginal Staphylococci strains isolated from the dam

To determine the relative importance of maternal microflora (faeces, vagina, and teats) in the contamination of newborn rats, strains of staphylococci from six different families (dam + litter) were isolated. These strains were identified, and by means of numerical profiles analyzed for their degree of similarity for each litter and (or) biotope. The staphylococci strains found in the gut of the newborn rat originated first from the teats and thereafter from the faeces. Concomitant observation of some identical strains, however, suggested a certain degree of similarity between these two maternal biotopes in this animal.     

生殖道支原体定植与阴道菌群结构的关联性

目的 探索女性性工作者生殖道中支原体定植与否与菌群结构的关联。方法 研究对象为2015年12月于云南红河州河口地区接受横断面调查的女性性工作者,采用PCR技术检测阴道中解脲脲原体（Uu）、人型支原体（Mh）和生殖支原体（Mg）的定植情况。将三种支原体任意一种检测阳性定义为支原体阳性,并依此将人群分为两组。在两组研究对象中各随机抽取9份样本行细菌16S rDNA基因的V3‒V4区高通量测序,通过生物信息学方法,比较两组研究对象阴道菌群结构差异。结果 支原体阳性组微生物群落的Shannon与Simpson指数均高于阴性组（Ps<0.05）,通过主成分分析图可以直观显示两组研究对象菌群结构具有明显差异,阳性组菌群结构更加多样化,没有集中趋势。支原体阳性组的乳杆菌属相对丰度低于阴性组（P<0.001）,而链球菌属（P=0.015）、普氏菌属（P=0.047）的相对丰度高于阴性组。结论 生殖道中支原体的定植与菌群结构有关,菌群结构的改变可能是支原体致病的重要条件之一。     

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora.     

Spatial relationship of the genital microflora to the vaginal epithelium of femal rats: transmission electron microscopy

Bacterial associations with vaginal epithelium were examined by transmission electron microscopy in castrate rats before and after estrogen treatment. Bacteria were associated with keratinized epithelium and appeared to be most abundant on exfoliated vaginal cells.     

Methods for quantitative and qualitative evaluation of vaginal microflora during menstruation.

The quantitative and qualitative changes in the bacterial flora of the vagina during menstruation have received inadequate study. Similarly, the effect of vaginal tampons on the microbial flora as well as the relationship between the microbial flora of the vagina and that of the tampon has not been adequately evaluated. The purposes of the present study were (i) to develop quantitative methods for studying the vaginal flora and the flora of tampons obtained during menstruation and (ii) to determine whether there were differences between the microflora of the tampon and that of the vaginal vault. Tampon and swab samples were obtained at various times from eight young healthy volunteers for 8 to 10 menstrual cycles. Samples consisted of swabs from women wearing menstrual pads compared with swab and tampon samples taken at various times during the menstrual cycle. Samples were analyzed for total facultative and anaerobic bacterial counts, and the six dominant bacterial species in each culture were identified. Statistical evaluation of the results indicates that total bacterial counts decreased during menstruation and that swab and tampon samples yielded similar total counts per unit weight of sample. The numbers of bacteria in tampons tended to be lower than in swabs taken at the same time. Overall, during menstruation, the concentrations of lactobacilli declined, but otherwise there was little difference among the species found during menstruation compared with those found in intermenstrual samples. Cotton tampons had little discernible effect on the microbial flora.     

Spatial relationship of the genital microflora to the vaginal epithelium of femal rats: transmission electron microscopy.

Bacterial associations with vaginal epithelium were examined by transmission electron microscopy in castrate rats before and after estrogen treatment. Bacteria were associated with keratinized epithelium and appeared to be most abundant on exfoliated vaginal cells.     

孕晚期GDM孕妇阴道菌群与阴道上皮细胞的

目的 探究孕晚期妊娠期糖尿病（GDM）孕妇阴道菌群与阴道上皮细胞的自噬和应激水平的相关性。方法 选取陆军军医大学第一附属医院2018年9月至2019年1月诊断及治疗的孕晚期GDM孕妇23例作为GDM组,选取同时段、同年龄段产检正常的孕妇23例作为对照组。对两组孕妇阴道菌群及自噬和应激水平进行检测,使用Pearson相关性分析探究菌群指标与自噬和应激水平间的相关性。结果 两组阴道细菌构成存在差异,GDM组在毛螺菌科、双歧杆菌目、普雷沃菌属、詹氏乳杆菌、双歧杆菌科、加德纳菌属的相对丰度大于对照组,对照组在乳杆菌属、卷曲乳杆菌、惰性乳杆菌、假单胞菌目、假单胞菌属、气球菌科的相对丰度大于GDM组。GDM组p62相对表达量显著低于对照组（t=3.903,P<0.001）,hsp70相对表达量显著高于对照组（t=5.704,P<0.001）。Pearson相关性分析结果显示,双歧杆菌目、加德纳菌属与p62水平呈显著负相关（r=-0.407、-0.482,P=0.009、0.002）,双歧杆菌科与hsp70呈显著正相关（r=0.448,P=0.004）;乳杆菌属与p62呈显著正相关（r=0.437,P=0.011）,与hsp70水平呈显著负相关（r=-0.475,P=0.005）。结论 孕晚期GDM孕妇阴道菌群与阴道上皮细胞自噬和应激水平存在一定相关性。     

Longitudinal study of the dynamics of vaginal microflora during two consecutive menstrual cycles

Background

Although the vaginal microflora (VMF) has been well studied, information on the fluctuation of the different bacterial species throughout the menstrual cycle and the information on events preceding the presence of disturbed VMF is still very limited. Documenting the dynamics of the VMF during the menstrual cycle might provide better insights. In this study, we assessed the presence of different Lactobacillus species in relation to the BV associated species during the menstrual cycle, assessed the influence of the menstrual cycle on the different categories of vaginal microflora and assessed possible causes, such as menstruation and sexual intercourse, of VMF disturbance. To our knowledge, this is the first longitudinal study in which swabs and Gram stains were available for each day of two consecutive menstrual cycles, whereby 8 grades of VMF were distinguished by Gram stain analysis, and whereby the swabs were cultured every 7^th day and identification of the bacterial isolates was carried out with a molecular technique.

Methods

Self-collected vaginal swabs were obtained daily from 17 non pregnant, menarchal volunteers, and used for daily Gram staining and weekly culture. Bacterial isolates were identified with tDNA-PCR and 16 S rRNA gene sequencing.

Results

Nine women presented with predominantly normal VMF and the 8 others had predominantly disturbed VMF. The overall VMF of each volunteer was characteristic and rather stable. Menses and antimicrobials were the major disturbing factors of the VMF. Disturbances were always accompanied by a rise in Gram positive cocci, which also appeared to be a significant group within the VMF in general.

Conclusions

We observed a huge interindividual variability of predominantly stable VMF types. The importance of Gram positive cocci in VMF is underestimated. L. crispatus was the species that was most negatively affected by the menses, whereas the presence of the other lactobacilli was less variable.     

Diethylstilbestrol-induced cervical and vaginal adenosis using the neonatal mouse model

The relevance of diethylstilbestrol (DES) administration to neonatal mice as a model for human pathology attributed to the use of DES in high-risk pregnancies has been investigated, particularly with respect to cervical and vaginal changes in female offspring. Neonatal DES treatment of mice results in tonic pituitary gonadotropin release and continuous estrogen secretion by the ovary. Studies were designed to determine the effect of this altered ovarian endocrine activity on cervical and vaginal histopathology. Ovariectomy of DES-treated mice, with or without estradiol replacement, did not eliminate the lesions, nor did estrogen and progesterone administered in a regimen intended to mimic estrous cycle changes. Induction of the constant estrus state by neonatal estradiol benzoate or testosterone propionate administration or by exposure to constant light did not produce the type of vaginal or cervical changes seen in DES mice. Thus, altered ovarian function is apparently not required for the vaginal and cervical changes appearing in later life. A role for endogenous (or exogenous) ovarian hormones in the developmental progression toward normality is suggested.     

Comparison between Gram stain and culture for the characterization of vaginal microflora: Definition of a distinct grade that resembles grade I microflora and revised categorization of grade I microflora

Background  

The microbiological diagnosis of bacterial vaginosis is usually made using Nugent's criteria, a useful but rather laborious scoring system based on counting bacterial cell types on Gram stained slides of vaginal smears. Ison and Hay have simplified the score system to three categories and added a fourth category for microflora with a predominance of the Streptococcus cell type. Because in the Nugent system several cell types are not taken into account for a final score, we carried out a detailed assessment of the composition of the vaginal microflora in relation to standard Gram stain in order the improve the diagnostic value of the Gram stain. To this purpose we compared Gram stain based categorization of vaginal smears with i) species specific PCR for the detection of Gardnerella vaginalis and Atopobium vaginae and with ii) tDNA-PCR for the identification of most cultivable species.