抗生素诱导小鼠阴道菌群失衡及乳杆菌调节作用的研究

目的 构建抗生素诱导的小鼠阴道菌群失衡模型,并研究乳杆菌对假丝酵母菌的阴道菌群失调的调整作用.方法 以IRC小鼠为实验对象,用头孢曲松造成阴道菌群失调,并将白假丝酵母接种到小鼠阴道内,造成小鼠阴道白假丝酵母过度增殖模型.对抗生素作用后的小鼠,采用乳杆菌预防、同步、治疗3种方式,阴道接种德氏乳杆菌DM8909菌液(2×108CFU/ml);取其阴道分泌物进行假丝酵母菌及乳杆菌的阴道菌群分析,并同时进行分泌物镜检,取阴道组织标本做病理组织学检查.结果 模型组小鼠阴道内细菌总数显著下降(P<0.01),出现阴道红肿、分泌物涂片镜检查到白细胞及假丝酵母.乳杆菌对小鼠抗生素导致的阴道菌群失衡的调节作用明显,受损小鼠阴道菌群的多样性和数量恢复至正常(P>0.05),其中乳杆菌预防组阴道菌群破坏程度最小(P<0.01);乳杆菌干预后的小鼠都可以显著性清除阴道内的假丝酵母菌(P<0.01),以乳杆菌预防组效果最好.结论 德氏乳杆菌DM8909能阻止白假丝酵母对阴道上皮的黏附作用,能调整阴道的菌群,对阴道黏膜有一定的修复功能.     

乳杆菌优良菌株的筛选及对小鼠阴道白假丝酵母菌过度增殖干预

目的探讨乳杆菌优良菌株的筛选及其对小鼠阴道白假丝酵母菌过度增殖干预作用。方法对德氏乳杆菌DM8909菌株进行诱变筛选,得到耐抗生素突变株208,以ICR小鼠为实验对象,用甲硝唑进行小鼠阴道菌群脱污染,将白假丝酵母菌接种到小鼠阴道内,构建小鼠阴道白假丝酵母菌过度增殖模型。采用阴道接种德氏乳杆菌DM8909突变株208菌液（2×108CFU/mL）处理,取其阴道冲洗液进行阴道菌群分析,并进行阴道上皮细胞的电镜检查。结果乳杆菌干预后显著清除阴道内的假丝酵母菌,肠杆菌恢复正常水平,与模型组比较,差异有统计学意义（P〈0.05）。结论德氏乳杆菌DM8909抗性菌株208能调整白假丝酵母菌在小鼠阴道内定植,对小鼠阴道上皮细胞有恢复作用。     

妊娠妇女阴道菌群的研究

本文对31例非妊娠健康妇女、31例早期妊娠妇女、31例中期妊娠妇女和31例晚期妊娠妇女的阴道菌群,就乳杆菌、酵母菌、葡萄球菌和肠杆菌进行定性、定量分析,同时测定阴道分泌物的PH。结果表明,妊娠妇女的阴道乳杆菌和酵母菌多于非妊娠妇女,而葡萄球菌和肠杆菌却少于非妊娠妇女。乳杆菌是阴道菌群中最重要的成员,妊娠时的生态优势地位更加突出。乳杆菌在阴道的生物屏障和酸性环境的维持方面起主要作用。乳杆菌可能是预防和治疗产道感染及新生儿感染的理想的生态制剂。     

先兆早产、胎膜早破、妊娠期糖尿病及正常妊娠女性阴道菌群分布的比较

目的:比较先兆早产、胎膜早破、妊娠期糖尿病及正常妊娠女性阴道菌群分布情况。方法:选择2016年6月至2018年6月在苏州大学附属第二医院妇产科住院的妊娠女性806例,其中先兆早产组206例,胎膜早破组234例,妊娠期糖尿病组156例,正常妊娠组210例。记录四组女性异常阴道菌群检出率及异常阴道菌群分布情况。结果:四组女性的年龄、孕周比较无统计学差异(P>0.05)。先兆早产组、胎膜早破组异常阴道菌群检出率高于妊娠期糖尿病组、正常妊娠组(P<0.05),而妊娠期糖尿病组、正常妊娠组异常阴道菌群检出率比较无统计学差异(P>0.05)。先兆早产组、妊娠期糖尿病组白色假丝酵母菌检出率高于胎膜早破组、正常妊娠组(P<0.05),先兆早产组、胎膜早破组阴道加德纳菌检出率高于妊娠期糖尿病组、正常妊娠组(P<0.05),先兆早产组无乳链球菌检出率高于胎膜早破组、妊娠期糖尿病组、正常妊娠组(P<0.05),胎膜早破组大肠埃希菌检出率高于先兆早产组、妊娠期糖尿病组、正常妊娠组(P<0.05)。结论:妊娠女性阴道感染以白色假丝酵母菌、大肠埃希菌、无乳链球菌、阴道加德纳菌为主,且先兆早产、胎膜早破女性阴道致病菌感染率较高,妊娠期糖尿病女性阴道白色假丝酵母菌的感染率较高。     

阴道常见乳杆菌及相关研究进展

近年来,随着全基因组测序等分子生物技术的应用,人们对阴道微生态系统的结构和功能的认识逐渐清晰。研究发现很多妇科疾病的出现伴随着阴道菌群失调,因此研究正常的阴道菌群结构和功能对预防和治疗菌群失调引起的疾病有重要意义,这也是目前人体微生态研究领域的热点。本文总结了近年关于健康女性阴道菌群的组成和类型的研究,阐述了阴道中常见乳杆菌如卷曲乳杆菌、惰性乳杆菌、格氏乳杆菌、詹氏乳杆菌等在功能研究方面的最新进展,以期为乳杆菌在防治妇科疾病如细菌性阴道病(BV)、外阴假丝酵母菌病(VVC)并推进其产品开发及临床应用等方面提供一些理论参考。     

不同生理阶段、避孕方式和感染状态对阴道乳酸杆菌影响的研究

目的调查不同生理阶段、避孕方式、感染因素对女性阴道乳酸杆菌数量的影响。方法收集南昌大学第二附院妇产科就诊的176例妇女的主要资料并采用qPCR方法定量分析阴道分泌物中乳酸杆菌数量,分泌物涂片和革兰染色后在显微镜下观察记录其感染状态。结果非绝经期组不同年龄段乳酸杆菌数量差异无统计学意义,绝经期组乳酸杆菌数量减少;不同避孕方式组的阴道乳酸杆菌数量差异无统计学意义;细菌性阴道病和假丝酵母菌感染组阴道乳酸杆菌数量较正常组差异均有统计学意义。结论生理阶段、细菌性阴道病和假丝酵母菌感染是影响女性阴道乳酸杆菌数量的主要因素。     

阴道白假丝酵母菌过度增殖的小鼠模型建立

目的建立阴道白假丝酵母菌过度增殖的小鼠模型。方法甲硝唑溶液（浓度为12．5mg／mL）30μL注入小鼠阴道内,1次／d,连续5d,白假丝酵母菌菌液（1×10^8CFU／mL）30μL接种到小鼠阴道内,1次／d,连用5d。取阴道冲洗液进行阴道菌群分析,并做阴道组织标本电镜观察。结果模型组小鼠阴道内白假丝酵母菌活菌数显著增加,乳杆菌活菌数量显著下降（P〈0．01）,出现阴道红肿、分泌物多和黏膜充血等白假丝酵母菌过度增殖的典型症状。结论通过抗生素脱污染后,小鼠阴道内接种白假丝酵母菌,能在小鼠阴道内定植,成功建立阴道白假丝酵母菌过度增殖的小鼠模型。     

外阴阴道假丝酵母菌病患者阴道菌群多样性研究

目的探讨外阴阴道假丝酵母菌病患者阴道菌群多样性的变化。方法采集育龄期女性健康受试者(control,CN,n=50)、细菌性阴道病患者(Bacterial vaginosis,BV,n=50)、外阴阴道假丝酵母菌病患者(Vulvovaginal candidiasis,VVC,n=48)、细菌性阴道病合并外阴阴道假丝酵母菌病患者(Bacterial vaginosis in combination of vulvovaginal candidiasis,BVC,n=32)的阴道拭子,提取细菌基因组DNA,而后应用25%~55%变性梯度的PCR-DGGE进行指纹图谱分析。结果阴道菌群指纹图谱结果显示,VVC组中阴道优势菌群与CN组基本一致,但与BV组和BVC组比较差异均有统计学意义。结论外阴阴道假丝酵母菌病阴道菌群多样性无明显变化。     

阴道乳杆菌胶囊调整产后阴道菌群失衡的临床研究

目的研究产后6~8周妇女阴道微生物环境的变化特征及阴道菌群失衡的发病率,探讨应用阴道乳杆菌胶囊恢复产后妇女阴道内环境,预防生殖道感染发生的临床意义。方法收集1 053例产妇产后6~8周回访就诊时阴道分泌物,采用革兰染色法检查,进行Nugent评分,按Nugent评分系统对菌群分类,进一步诊断细菌性阴道病(BV),并与其孕早期产检时情况比对。结果 1 053例产后6~8周妇女Nugent评分正常521例,临界242例,有290例发生阴道炎症,细菌性阴道病205例,阴道假丝酵母菌74例和滴虫11例。疾病构成比以非特异性阴道炎为主,特异性阴道炎较为少见。所有妇女阴道p H均在5.0以上。在所有涂片中,被认为在维持正常阴道微生态平衡中起关键作用的乳杆菌数量和比例明显不足。产后BV的发病率(31.36%)高于孕早期产检时,产后妇女BV发病率较高,差异有统计学意义(P0.05)。予300例患者阴道内放置乳酸杆菌活菌胶囊,结果显示研究组阴道p H明显降低,接近正常女性阴道p H水平(P0.05),且阴道乳酸杆菌数量明显增多(P0.05);同时产后3个月、6个月阴道炎的发生率均明显低于对照组(P0.05)。结论产后妇女阴道乳酸杆菌量减少,菌群比例明显失调。阴道炎症以非特异性阴道炎为主,细菌性阴道病、真菌性阴道炎和滴虫性阴道炎较少见。乳杆菌胶囊作为生态治疗的活菌制剂,既能预防、治疗阴道感染,又无不良反应,还可以减少产后妇女阴道炎的复发率,值得临床推广应用。     

乳杆菌代谢物脂肪酸组分对白假丝酵母菌的抑制及临床疗效观察

目的观察乳杆菌DM9811代谢物中脂肪酸组分（以下称抑菌液）在体内外对白假丝酵母菌的抑制效果,探索外阴阴道假丝酵母菌病（VVC）的安全、有效、无副作用的生态制剂疗法。方法不同浓度抑菌液对白假丝酵母菌的作用及作用不同时间的抑菌效果;对2003年9月～2004年1月57例VVC患者分别给予抑菌液（试验组）及达克宁栓（对照组）进行治疗,并做相关细菌培养及疗效判定。结果（1）体外试验中抑菌液对白假丝酵母菌的抑制作用随着浓度的降低而降低;在1：2浓度（乙酸含量0．8mg／ml）抑制作用随着时闻的延长而增强,12h达100％;对乳杆菌无抑制作用。（2）体内试验中抑菌液使阴道白假丝酵母菌数量明显减少（P〈0．05）,使阴道pH明显降低（P〈0．05）,阴道内乳杆菌数量差异无显著性（P〉0．05）;达克宁栓使阴道内白假丝酵母菌数量明显减少,乳酸杆菌数量明显下降（P〈0．05）,而阴道pH无变化。（3）2组1疗程治愈率及1个月复查治愈率均差异无显著性。（4）副反应：两者比较差异有显著性（P〈0．05）。结论抑菌液在体内外对白假丝酵母茵均有明显的抑杀作用;降低阴道pH;对阴道乳杆菌数量无影响;无副作用;可用于VVC的治疗。     

Analysis of vaginal lactic acid producing bacteria in healthy women

Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.     

健康妊娠妇女阴道乳酸杆菌及pH变化的研究

目的研究健康妊娠妇女阴道乳酸菌数量及pH的变化与妊娠时间的关系。方法选择20例健康非妊娠妇女,60例健康妊娠妇女(其中早期妊娠、中期妊娠及晚期妊娠各20例)阴道分泌物进行乳酸杆菌数量检测及pH测定,并对乳酸杆菌产生过氧化氢的情况进行检测。结果随着妊娠时间的增加,乳酸杆菌的数量明显增高,伴随阴道分泌物的pH逐渐降低。结论乳酸杆菌与健康妊娠妇女阴道的生物屏障和酸性环境的维持有重要关系。     

The correction of dysbiotic disorders of the vaginal microflora by using a preparation made from highly adhesive lactobacteria

The effectiveness of a new bacterial preparation obtained from highly adhesive lactobacteria and intended for the correction of dysbiotic disturbances of vaginal microflora was studied in the treatment of 60 pregnant women with dysbacteriosis of the maternal passages. 30 pregnant women were simultaneously treated by the vaginal application of Lactobacterin. The study showed that the use of the preparation of highly adhesive lactobacteria caused the pronounced and stable correction of the microflora of the maternal passages. This correction was manifested by the domination of lactic acid bacterial flora and a decrease in the number of opportunistic microorganisms.     

多次人工流产后妊娠期阴道病患者致病微生物检测及药敏分析

目的检测多次人工流产后妊娠期阴道病患者的致病微生物,并提出治疗建议。方法选取102例多次人工流产后妊娠期阴道病患者作为研究对象,所有患者均采用无菌棉拭子收集2份阴道分泌物样本,其中1份采用高倍镜检查阴道清洁度和滴虫感染情况,另1份检测病原微生物感染情况,统计病原微生物分布情况并检测常见病原微生物药敏情况。结果阴道清洁度检测显示,清洁度4度者构成比最高,为60.78%,其次为3度,1度者构成比最低。共12例患者存在滴虫感染,构成比为11.76%。人乳头瘤病毒(HPV)感染12例,感染率为11.76%。支原体感染率、衣原体感染率分别为38.24%、33.33%。92例患者阴道分泌物共检出225株病原菌,其中革兰阳性菌占60.44%,以棒状杆菌、金黄色葡萄球菌、表皮葡萄球菌为主;革兰阴性菌占25.78%,以肺炎克雷伯菌、阴道加德纳菌、大肠埃希菌为主;真菌占13.78%,以白假丝酵母、光滑假丝酵母、克柔假丝酵母为主。药敏试验显示,棒状杆菌、金黄色葡萄球菌、表皮葡萄球菌、乳杆菌和肠球菌属对青霉素的耐药率均较低,分别为0.00%、0.00%、5.00%、5.88%、6.67%;肺炎克雷伯菌、阴道加德纳菌、大肠埃希菌对亚胺培南的耐药率均较低,分别为4.55%、12.50%、9.09%;白假丝酵母、光滑假丝酵菌、克柔假丝酵母对两性霉素B、5-氟胞嘧啶的耐药率均为0.00%,且光滑假丝酵母菌对酮康唑、克霉唑的耐药率也均为0.00%。结论多次人工流产后妊娠期阴道病患者常见的致病微生物有滴虫、支原体、衣原体、细菌、真菌等,需要根据检测结果选择安全、合理的药物进行治疗。     

Diagnostic value of Gram stain for assessment of vaginal smears during pregnancy

The main aim of this study was to compare the diagnostic value of Giemsa stained method with Gram stained method for the evaluation of vaginal smears among pregnant women. A study population comprised 111 pregnant between 6 and 30 weeks of gestation. The vaginal smears from every subject was diagnosed according to Giemsa and Gram stained method and micro-organisms were isolated by culture. In 29.3% cases diagnosed as normal flora (2a) on the basis of Giemsa method bacterial vaginosis was detected in Gram stains according to Spiegel's criteria and pathological microflora in concentration > or = 10(5) CFU/ml was cultured among 75.9% of them. Among 31.7% women who had grade 3a (abnormal) in Giemsa stains method normal flora was diagnosed on the basis on Gram's method and from 17.1% pregnant women from this group we did not isolated any pathogens. For evaluation of vaginal smears during pregnancy the Giemsa method should be replaced by Gram stained method.     

Inhibitory activity of vaginal Lactobacillus bacteria on yeasts causing vulvovaginal candidiasis

Growing frequency of therapeutical failures of vulvovaginal candidiasis, resulting from resistance of certain species of Candida to imidazole agents, raises interest in the use of probiotics from Lactobacillus genera as prophylaxis. Unfortunately, little is known about inhibitory mechanisms of Lactobacillus on Candida. The aim of this study was to compare the activity of selected Lactobacillus species, representing the physiological vaginal flora, against Candida as well as investigation whether their inhibitory activity against Candida is related strictly to hydrogen peroxide and lactic acid production. 125 strains from vaginal smears of healthy women were classified by making use of phenotypic and genotypic methods. The majority of strains belonged to L. acidophilus: L. acidophilus sensu stricto, L. crispatus, L. gasseri and L. johnsonii as well as L. fermentum and L. plantarum species. Culture supernatants of selected 25 strains representing the isolated species were examined for their inhibitory activity against the growth of Candida albicans and C. glabrata. The results showed that the strongest and the fastest activity against C. albicans was demonstrated by L. delbrueckii strains, producing the largest quantities of hydrogen peroxide. On the other hand, extended activity, demonstrable after 24 hours, was shown by non-H2O2 producing L. plantarum supernatants. Growth of C. glabrata was not inhibited by any of the examined strains of Lactobacillus. Comparison of activity of live active cultures of Lactobacillus strains and their mixtures with this of pure H2O2 and lactic acid has shown that pure chemical compounds were less active than the cultures. This suggests that mixtures of Lactobacillus strains are in cooperation with each other using many different metabolites.     

An Evaluation of Risk Factors in Pregnant Women with <Emphasis Type="Italic">Candida</Emphasis> Vaginitis and the Diagnostic Value of Simultaneous Vaginal and Rectal Sampling

In this study, we investigated the epidemiological characteristics of VVC among pregnant women. We conducted a prospective survey among 372 pregnant women to investigate the prevalence, clinical forms, etiological agents, and predisposing factors of VVC. In addition, we determined the relationship between vaginal and rectal flora by simultaneously obtaining one high vaginal swab and one rectal swab from each patient using sterile cotton-tipped swabs. Furthermore, we compared the recovery and identification performances of chromID Candida agar to Sabouraud dextrose agar with gentamicin and chloramphenicol. Clinically and mycologically confirmed cases of VVC were detected in 139 (37.4%) and vaginal colonization described in 42 (11.3%) of 372 pregnant women. Rectal cultures were also positive in 98 of the 139 (70.5%) VVC cases. Candida albicans and C. glabrata were identified in vaginal samples in 58.0 versus 19.0% and from rectal samples in 49.0 versus 13.5%, respectively. Increases in gestational week and gravidae were identified to be statistically significant in patients with acute VVC (AVVC) and symptomatic recurrent VVC (RVVC), and asymptomatic RVVC (P = 0.04 and P = 0.03, respectively). In the laboratory diagnosis of VVC, specifically tailored chromogenic media are reliable tools for both the recovery and rapid identification of common Candida spp., particularly C. albicans, as well as for the detection of polyfungal populations in vaginal samples (P > 0.05). In addition, rectal colonization is a common finding in cases of AVVC and symptomatic-RVVC cases and corresponds well with the presence of the same yeast species in the vagina.     

妇科门诊育龄妇女与孕妇阴道菌群状态及常规感染的比较分析

目的比较妇科门诊育龄妇女与孕妇的阴道菌群状态及常规感染,分析阴道菌群改变对阴道常规感染影响。方法用革兰染色法分别检查普通育龄妇女和孕妇的阴道分泌物,并进行Nugent评分。结果妇科门诊育龄妇女和孕妇的假丝酵母菌阳性率分别为28．6％和9．2％,滴虫阳性率分别为2．4％和0．6％;细菌性阴道病（BV）阳性率分别为32．4％和9．6％;乳杆菌3＋到4＋的比例分别为74．4％和44．8％。2组之间比较差异均有显著性（P〈0．05）。结论相对于妇科门诊育龄妇女而言,孕妇的阴道微生态状况更健康,对各种病原微生物有更强的抵抗力,受假丝酵母菌、滴虫和细菌性阴道病等常规疾病感染的机会明显降低。