Objective:To study the growth characteristics of Lactobacillus and Enterococcus faecalis co-cultured in vaginal flora, and to explore the effect of the supernatant of Lactobacillus and Enterococcus faecalis co-cultured on the formation of Gardnerella vaginalis biofilm.Methods:Enterococcus faecalis and Gardner vaginalis isolated, purified and identified from vaginal secretions of patients with clinical diagnosis of BV,and Lactobacillus curlicus isolated, purified and identified from vaginal secretions of patients with abnormal flora diagnosed in the gynecological vaginal microecology evaluation system.The growth and lactic acid production ability of Lactobacillus and Enterococcus faecalis co-cultured for 24h, 48h and 72h were observed.Observe the biofilm formation of Gardnerella vaginalis after being cultured alone and co-cultured with the supernatant of Lactobacillus, Enterococcus faecalis and their mixed cultures for 24h, 48h and 72h by detecting the OD value.Results:After mixed culture of Lactobacillus and Enterococcus faecalis, the production of L-lactic acid was less than that of Lactobacillus alone and Enterococcus faecalis alone（P＜0.05）. With the increase of time, the number of bacteria co-cultured with Lactobacillus and Enterococcus faecalis increased gradually, but more than that of Enterococcus faecalis alone（P＜0.05）. With the increase of time, the film forming ability of co-culture of Lactobacillus and Enterococcus faecalis was better than that of single culture of Lactobacillus and Enterococcus faecalis（P＜0.05）.At 24h, the supernatant of Lactobacillus had no significant inhibition on the membrane forming ability of Gardner bacteria, while the supernatant of Enterococcus faecalis and the supernatant of co-culture of Lactobacillus and Enterococcus faecalis had significant inhibition on the biofilm formation of Gardner bacteria（P＜0.05）.At 48h and 72h, the supernatant of Lactobacillus, Enterococcus faecalis, and the supernatant co-cultured with Lactobacillus and Enterococcus faecalis had a strong inhibitory effect on the membrane forming ability of Gardnerella, and the supernatant co-cultured had a stronger inhibitory effect than the supernatant cultured alone, with a statistically significant difference（P＜0.05）.Conclusions:Lactobacillus and Enterococcus faecalis co-culture have antagonistic effect on lactic acid production, and co-culture can promote the formation of biofilm, but not the growth of bacteria.With the increase of culture time, the supernatant of Lactobacillus and Enterococcus faecalis co-culture has a stronger inhibitory effect on Gardnerella vaginalis biofilm than that of Lactobacillus or Enterococcus faecalis, indicating that Lactobacillus and Enterococcus faecalis co-culture have a synergistic effect on Gardnerella vaginalis biofilm inhibition.

目的 分析稽留流产患者阴道菌群情况.方法 选取 2020年 9月至 2021年 6 月首都医科大学附属北京妇产医院就诊的患者,将稽留流产患者 30 例纳入试验组,正常早孕拟行人工流产患者 30 例纳入对照组,并将稽留流产患者进一步根据胚胎绒毛染色体检查结果分为染色体正常组 16 例、染色体异常组 6 例、染色体不详组8 例,填写调查问卷,行阴道分泌物微生态检测,并将四组基线资料、阴道微生态指标进行单因素比较,同时采用16S rRNA高通量测序技术分析四组患者阴道菌群的差异.结果 四组基线资料与微生态指标比较结果显示,四组既往是否有盆腔炎病史、菌群多样性、优势菌比较,差异有统计学意义(P＜0.05),其中染色体异常组既往盆腔炎病史比例、初次性生活年龄高于对照组(P＜0.05),而既往流产史比例低于对照组(P＜0.05).OTU聚类分析结果显示,相较于稽留流产各组,对照组OTU数目更多,菌群更丰富;物种分类分析结果显示,与对照组比较,染色体正常组存在不同乳杆菌菌株比例失调(惰性乳杆菌增加,未分类乳杆菌减少)及除乳杆菌以外的其他细菌(加德纳菌、普雷沃菌、埃希氏-志贺菌等)相对丰度增加,而染色体异常组在乳杆菌丰度及菌株比例比较,差异无统计学意义(P＞0.05);Alpha多样性分析结果显示,四组菌群丰度比较,差异有统计学意义(P＜0.05),其中染色体正常组菌群丰度低于对照组(P＜0.05),染色体异常组与对照组菌群丰度比较,差异无统计学意义(P＞0.05);β多样性分析及线性判别分析结果显示,四组阴道菌群比较,差异无统计学意义(P＞0.05).结论 稽留流产患者存在阴道菌群失衡,可能与阴道菌群中存在乳杆菌相对丰度降低和/或不同乳杆菌菌株比例失衡及除乳杆菌以外的其他细菌(加德纳菌、普雷沃菌、埃希氏-志贺菌等)相对丰度增加有关,但其直接病因及机制有待进一步研究.

目的:研究卷曲乳杆菌与植物乳杆菌联合共培养后对白假丝酵母菌抑制作用的差异.方法:选取首都医科大学附属北京妇产医院就诊患者的阴道分泌物,通过分离纯化鉴定筛选得到2 株卷曲乳杆菌(乳杆菌A、B)和1 株植物乳杆菌(乳杆菌C).比较乳杆菌A、B、C单独作用时对10 株白假丝酵母菌临床株的抑制作用差异.比较乳杆菌A与C、B与C按1 ∶ 1 混合共培养后对10 株白假丝酵母菌临床株的抑制作用差异.比较混合乳杆菌与单一乳杆菌对10 株白假丝酵母菌临床株的抑制作用差异.结果:单一乳杆菌A、B对白假丝酵母菌(实验浓度)的抑制作用差异无统计学意义(P>0.999).乳杆菌C对白假丝酵母菌的抑制作用明显优于单一乳杆菌A、B,差异有统计学意义(P<0.001,P<0.001).混合乳杆菌A+C(1 ∶ 1)、乳杆菌B+C(1 ∶ 1)对白假丝酵母菌的抑制作用明显优于乳杆菌A、B、C任意单一菌,差异均有统计学意义(P<0.001、P<0.001、P=0.016;P<0.001、P<0.001、P=0.018).混合乳杆菌A+C(1 ∶ 1)与乳杆菌B+C(1 ∶ 1)对不同白假丝酵母菌的抑制作用差异无统计学意义(P>0.999).结论:对于单菌来说,植物乳杆菌C对白假丝酵母菌的抑制效果优于卷曲乳杆菌A、B.对于混合菌,即乳杆菌A+C或乳杆菌B+C按1:1 混合,对白假丝酵母菌的抑制效果明显优于乳杆菌A、B、C单独作用.

目的:研究阴道菌群中粪肠球菌上清液对阴道加德纳菌、阴道阿托波菌和两种菌混合生长的生物膜形成的影响.方法:选取初次诊断为BV患者阴道分泌物中分离纯化鉴定的阴道加德纳菌和阴道阿托波菌,观察加德纳菌单独培养及加德纳菌与阿托波菌共培养24、48、72 h的生长情况,通过检测OD值观察生物膜形成情况.通过检测OD值观察加德纳菌单独培养及其与阿托波菌共培养时加入含粪肠球菌代谢产物的上清液24、48、72 h的生物膜形成情况.结果:加德纳菌生物膜形成较好,阿托波菌几乎无生物膜形成.总菌量相等的加德纳菌单独培养与加德纳菌和阿托波菌共培养时的OD值无显著差异(P=0.259,P>0.05).加德纳菌及其与阿托波菌共培养时加含粪肠球菌代谢产物的上清液较不加上清液时OD值显著降低(P<0.05).结论:加德纳菌和阿托波菌有相互促进生物膜形成作用.加入含粪肠球菌代谢产物的上清液对加德纳菌和加德纳菌与阿托波菌共培养形成的生物膜有显著抑制作用.