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。     

复发性外阴阴道念珠菌病的阴道微生态特征分析

目的拟从阴道菌群的微生态特征入手,旨在探讨阴道微生物群落结构与外阴阴道念珠菌病(Vulvovaginal Candidiasis,VVC)发病的潜在联系。方法 2013年1月~2014年12月,126名处于VVC急性发作且以往有RVVC史的非孕育龄妇女作为研究对象,另有120名正常育龄期体检女性作为对照组。测定阴道p H值,并采集阴道分泌物行革兰染色镜检,行Nugent评分。同时,通过阴道菌群密集度、菌群多样性、优势菌等指标对阴道微生态进行评价。结果患者组的阴道p H值(4.51±0.3 Vs 4.11±0.2,P0.05)和Nugent评分(4.21±0.81 Vs 1.31±1.19,P0.05)显著高于对照组。患者组乳酸杆菌密集度显著低于对照组,而阴道加特纳菌/普雷沃菌、革兰阳性球菌和其他细菌的密集度显著高于对照组(P0.05)。同时,患者组菌群多样性显著高于正常组;且患者组乳酸杆菌优势比例显著低于对照组(55.56%Vs 95.00%,P0.05),而阴道加特纳菌/普雷沃菌(23.02%Vs 1.67%,P0.05)和革兰阳性球菌优势比例显著高于对照组(17.46%Vs 1.67%,P0.05)。结论RVVC患者阴道微生态特征与正常育龄妇女存在显著差异,生物屏障的削弱可能是VVC反复发作的风险因素。新型阴道乳酸菌制剂的研发和应用可能在重建阴道微生态,预防VVC复发方面获得突破。     

细菌性阴道病患者阴道分泌物16SrDNA序列分析

目的分析健康妇女及细菌性阴道病(Bacterial vaginosis,BV)患者阴道分泌物16S rDNA序列。方法提取20例健康妇女及40例BV患者阴道分泌物标本中的总DNA,针对细菌16S rDNA保守区设计通用引物进行PCR扩增、克隆、测序,将获得的16S rDNA序列与美国国立生物技术信息中心(NCBI)数据库中的发表序列进行比对,分析克隆群中细菌种类和比例。结果通过阴道分泌物16S rDNA序列分析,发现健康妇女阴道分泌物中以卷曲乳酸杆菌(Lactobacillus crispatus),惰性乳酸杆菌(Lactobacillus iners),加氏乳酸杆菌(Lactobacillus gasseri)为优势菌种,而BV患者阴道菌群种类繁多,以加德纳菌属(Gardnerella)和奇异菌属(Atopobium vaginae)克隆子占较大比例,仅4例患者可见卷曲乳酸杆菌(Lactobacillus crispatus),其他患者均未见有乳酸杆菌克隆子且奇异菌属阴道病患者甲硝唑治疗疗效较差。结论健康妇女和BV患者阴道分泌物菌群种类有较大区别,BV患者在治疗前进行16S rDNA序列分析检测有较大的临床意义。     

基于高通量测序分析健康人 咀嚼槟榔前后口腔菌群多样性

目的为揭示健康人咀嚼槟榔前后的菌群结构及其多样性,分析咀嚼槟榔前、咀嚼5 min后和咀嚼30min后口腔内菌群结构变化以及多样性特征。方法通过收集8个人咀嚼槟榔前后唾液,用宏基因组DNA进行高通量测序。结果咀嚼槟榔5min后,链球菌属(Streptococcus)、普雷沃菌_7属(Prevotella_7)、韦荣球菌属(Veillonella)、纤毛菌属(Leptotrichia)、嗜血杆菌属(Haemophilus)、拟普雷沃菌属(Alloprevotella)、普雷沃菌属(Prevotella)、卟啉单胞菌属(Porphyromonas)、梅毒螺旋体_2(Treponema_2)、普雷沃菌_6属(Prevotella_6)和兼性双球菌属(Gemella)较咀嚼前相对丰度明显下降,分别下降了16.57%、12.94%、9.38%、23.08%、54.55%、52.63%、30.00%、42.86%、30.00%、16.67%和42.86%,纤毛菌属(Leptotrichia)降幅最大。而咀嚼30min后,纤毛菌属(Leptotrichia)、嗜血杆菌属(Haemophilus)、拟普雷沃菌属(Alloprevotella)、卟啉单胞菌属(Porphyromonas)、梅毒螺旋体_2(Treponema_2)和罗思菌属(Rothia)的相对丰度较咀嚼5 min后均增大。结论链球菌属(Streptococcus)、韦荣球菌属(Veillonella)作为主要变化的菌属,在咀嚼槟榔的过程中,对口腔产生一定的作用,可能是口腔龋齿等牙周疾病的直接或间接影响因子。     

细菌性阴道病的诊治进展

正常阴道内有需氧菌和厌氧菌寄居形成阴道正常菌群．阴道与这些菌群之间形成生态平衡．因此并不致病。一旦生态平衡被打破或外源性病原体侵入,即可导致炎症的发生。长期应用抗生素抑制乳酸杆菌的生长或机体免疫力低下亦可使其他致病菌成为优势菌,引起炎症。大量研究表明。细菌性阴道病(曾被命名为嗜血杆菌性阴道炎)已成为生育期妇女发病率最高的阴道感染性疾病。     

细菌性阴道病微生态与益生菌

细菌性阴道病(BV)是育龄妇女阴道炎的最常见原因,其特征是栖居在阴道内的乳酸杆菌减少导致阴道菌群平衡失调的复杂变化。用乳酸杆菌替代抗生素是治疗BV的一种有效的治疗措施。乳酸杆菌产生过氧化氢、乳酸、杆菌素能抑制引起BV的细菌生长。多胺在BV的病理机制中有重要作用。本研究拟综述BV患者阴道菌群的变化,免疫防御反应,阴道乳酸杆菌对BV的影响,多胺的意义,益生菌在治疗选择中的作用。     

1500例女性阴道微生态状况分析

目的女性阴道微生态状况调查与分析。方法采用阴道pH、阴道涂片Gram染色镜检(菌群密集度、菌群多样性、优势菌、真菌孢子、滴虫和炎性反应等)、阴道微生物功能测定,评估1 500例体检者阴道微生态状况。结果微生态正常830例(55.7%),微生态失调者670例(44.6%)。在670例微生态失调患者中,需氧菌阴道炎156例(23.3%),细菌性阴道病265例(39.6%),外阴阴道假丝酵母菌病31例(4.63%),滴虫性阴道炎2例(0.29%),混合感染216例(32.2%)。结论阴道微生态评价有助于筛查无症状阴道微生态失调患者,对指导临床评价感染及预防阴道感染具有重要的临床意义。     

阴道炎患者乳酸杆菌数量变化与临床的研究

目的探讨阴道炎患者的阴道乳酸杆菌数量的变化,以指导临床治疗。方法用LBS培养基分离乳酸杆菌,观察阴道炎患者乳酸杆菌变化的情况。结果172例阴道炎病例中,乳酸杆菌数量平均数明显减少。结论乳酸杆菌数量的减少、阴道菌群的改变是阴道炎发病的重要原因,重新构建阴道正常菌群是阴道炎治疗的重点。     

唾液菌群特征分析对儿童龋齿复发的预测价值

目的探究唾液菌群特征对儿童龋齿复发的预测价值。方法收集2017年1月至2018年1月在中铁十七局集团有限公司中心医院诊断及治疗的龋齿患儿94例,在进行龋齿治疗后第4周搜集唾液样本,并根据12个月后的龋齿发生情况分为复发组及未复发组,对比两组微生物检测结果,并使用ROC曲线评估微生物指标对龋齿复发的预测效能。结果本研究随访结束时,龋齿复发23例(25.84%)。龋齿复发与未复发组的唾液微生物组成存在显著差异:复发组的变形链球菌(Streptococcus mutans)、内氏放线菌(Actinomyces naeslundii)、普雷沃氏菌属(Prevotella)、二氧化碳噬纤维菌属(Capnocytophaga)、普雷沃氏菌科(Prevotellaceae)、放线菌属(Actinobacillus)的相对丰度显著高于未复发组(均P<0.05),未复发组在纤毛杆菌属(Leptotrichia)、奈瑟氏菌属(Neisseria)、链球菌属(Streptococcus)的相对丰度显著高于复发组(均P<0.05)。纤毛杆菌属、内氏放线菌、普雷沃氏菌属、变形链球菌载量预测龋齿复发的AUC值分别为0.753、0.715、0.680、0.940,其中变形链球菌的AUC值显著高于其他三种菌属(均P<0.05)。结论分析唾液菌群对儿童龋齿的复发具有一定的预测价值,其中变形链球菌的预测效能较好。     

基于实时定量PCR技术的育龄期细菌性阴道病患者 阴道菌群结构分析及其临床应用

摘要：目的 探讨育龄期女性细菌性阴道病（bacterial vaginosis,BV）患者阴道优势菌群变化及其与临床指标的相关性。方法 选择本院门诊确诊的育龄期BV患者35例和同期入院体检的健康体检女性37例,无菌拭子采集阴道中段壁分泌物,提取细菌基因组DNA,采用实时定量PCR技术（real-time quantitative PCR,qPCR）进行阴道优势细菌检测,并进行其与临床指标如阴道pH和Nugent评分相关性分析。结果 乳杆菌属细菌及其种水平的细菌如惰性乳杆菌、卷曲乳杆菌、詹氏乳杆菌等在BV患者中均显著下降,BV相关阴道致病细菌如加德纳菌属、奇异菌属、埃格特菌属、巨型球菌I型菌属、纤毛菌属和普氏菌属显著升高（P<0.05）。阴道致病菌群与阴道pH和Nugent评分呈显著正相关,而阴道卷曲乳杆菌和惰性乳杆菌与其呈负相关。结论 育龄期BV患者阴道优势菌群显著失衡,并与阴道pH和Nugent评分显著相关,提示阴道优势菌群改变参与BV发生发展。     

Characterization of some strains from human clinical sources which resemble "Leptotrichia sanguinegens": description of Sneathia sanguinegens sp. nov., gen. nov

Three strains of a gram-negative, blood or serum requiring, rod-shaped bacterium recovered from human clinical specimens were characterised by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing showed the unknown rod-shaped strains are members of the same species as some fastidious isolates recovered from human blood specimens and previously designated "Leptotrichia sanguinegens". Based on phylogenetic and phenotypic evidence, it is proposed that the isolates from human sources be classified in a new genus Sneathia, as Sneathia sanguinegens gen. nov., sp. nov. The type strain of Sneathia sanguinegens is CCUG 41628T.     

不同年龄组阴道炎患者阴道分泌物检查结果分析

目的对比分析不同年龄组女性阴道炎患者阴道清洁度和霉菌、滴虫、纤毛菌、线索细胞检出情况。方法对2 295例女性阴道炎患者按照年龄分组进行阴道分泌物涂片染色检查。结果 2 295例中阴道清洁度为Ⅲ～Ⅳ度占66.19%,霉菌、线索细胞、滴虫和纤毛菌的检出率分别为25.22%、15.69%、0.74%和0.48%;50岁以上年龄组阴道清洁度Ⅲ度检出率较高;20岁以下年龄组Ⅳ度检出率较高;21～30年龄组霉菌检出率较高,41～50年龄组线索细胞和滴虫检出率较高,纤毛菌检出率在各组之间差异无统计学意义。结论大连地区女性阴道炎患者防治形势不容乐观;与年龄相关的生理原因及个人行为因素导致不同年龄组患者阴道感染病因的差异性。     

Study of acute vulvovaginitis in sexually active adult women, with special reference to candidosis, in patients of the Francisco J. Muñiz Infectious Diseases Hospital

The results of microbiological vaginal secretions samples obtained from 749 women (from July 2001 to July 2002) were studied in the Bacteriology Unit of the Francisco Javier Mu?iz Hospital from Buenos Aires. All patients suffered acute vulvovaginitis were child bearing and sexually active women, 334 of them were HIV-positive. The following are the results of the microbiological studies: Lactobacillus spp 50.6%, Gardnerella vaginalis 25.6%, Candida spp 17.4%, Trichomonas vaginalis 5.3%, Neisseria gonorrhoeae 0.3% and B group Streptococcus 0.8%. Candida vaginitis was significantly more frequent in HIV-positive patients, (21.6% vs 14%; p = 0.0086); meanwhile, trichomoniasis was less common although the difference was not statistically significant (3.6 vs 6.7%, p = 0.0810). The following Candida species were isolated in this study: Candida albicans 76.8%, Candida glabrata 15.6%, Candida parapsilosis 2.9%, Candida tropicalis 1.5% and Candida krusei 0.7%. Eight cases (6.2%) of vaginitis were produced by two Candida species (C. albicans and C. glabrata), and in three cases (2.17%) Saccharomyces cerevisiae were isolated. Five women suffering acute vaginitis with Candida spp presented another etiologic agent of vaginal infection, three cases T. vaginalis and two cases G. vaginalis. The following are some of the most important findings of this study: 1) Half of the patients presented a normal microbial biota; 2) Candida spp vaginitis was significantly more frequent among HIV-positive women; 3) we observed a high incidence of Candida glabrata infections (15.9%), 4) 6.2% of vaginal candidiasis were caused by more than one Candida species and, 5) the susceptibility pattern of C. albicans and C. glabrata isolates against fluconazole was similar to the one observed in other studies. The majority of C. albicans isolates were susceptible to fluconazole (MIC90 = 0.5 microg/ml) meanwhile C. glabrata strains were much less susceptible to this drug (MIC50 and MIC90 = 32 microg/ml).     

The range of antagonistic effects of Lactobacillus bacterial strains on etiologic agents of bacterial vaginosis

Bacterial vaginosis is caused by uncontrolled sequential overgrowth of some anaerobic bacteria: Gardnerella vaginalis, Prevotella bivia, Bacteroides spp., Peptostreptococcus spp., Mobiluncus sp. usually occurring in stable numbers in the bacterial flora of healthy women. On the other hand, different species of bacteria belonging to the genus Lactobacillus, most frequently L. plantarum, L. rhamnosus and L. acidophilus, form a group of aerobic bacteria dominating in the same environment. The diversity and density of their populations depend on the age and health conditions. Thanks to their antagonistic and adherence properties bacteria of the genus Lactobacillus can maintain a positive balance role in this ecosystem. The aim of this study was to assess the antagonistic properties of Lactobacillus strains isolated from the vagina of healthy women against most common agents of bacterial vaginosis. It was found that nearly all of the tested Lactobacillus strains exerted distinct antagonistic activity against anaerobic bacteria: Gardnerella vaginalis, Prevotella bivia and Peptostreptococcus anaerobius and quite a number also against Gram-negative rods, while only some of them were able to inhibit Gram-positive aerobic cocci as Enterococcus faecalis or Staphylococcus aureus.     

Vaginitis: current microbiologic and clinical concepts.

Infectious vaginitis occurs when the normal vaginal flora is disrupted; it may arise when saprophytes overwhelm the host immune response, when pathogenic organisms are introduced into the vagina or when changes in substrate allow an imbalance of microorganisms to develop. Examples of these types of vaginitis include the presence of chronic fungal infection in women with an inadequate cellular immune response to the yeast, the introduction of trichomonads into vaginal epithelium that has a sufficient supply of glycogen, and the alteration in bacterial flora, normally dominated by Lactobacillus spp., and its metabolites that is characteristic of "nonspecific vaginitis". The authors review microbiologic and clinical aspects of the fungal, protozoal and bacterial infections, including the interactions of bacteria thought to produce nonspecific vaginitis, that are now recognized as causing vaginitis. Other causes of vaginitis are also discussed.     

Anaerobic bacteria cultured from the tongue dorsum of subjects with oral malodor

The bacteria on the dorsum of the tongue are the most frequent cause of oral malodor; however, the bacterial flora of the tongue has not been well defined. Although recent studies have used DNA probes to detect the presence of certain periodontal pathogens, cultural studies have been limited because of the complexity of the flora of the tongue dorsum. The purpose of this study was to grow and to identify maximum numbers of capnophylic Gram-negative bacilli and anaerobic micro-organisms by culturing tongue samples on to several selective and non-selective media. The most frequently isolated species included Peptostreptococcus anaerobius, Collinsella aerofaciens, Eubacterium group, Actinomyces spp., Eikenella corrodens, Veillonella spp., Fusobacterium nucleatum, pigmented Prevotella spp. and Selenomonas spp. Reported for the first time are Actinomyces turicensis, Collinsella aerofaciens, Eubacterium saburreum, E. timidum, Prevotella tannerae, Campylobacter concisus, Campylobacter mucosalis, Leptotrichia buccalis, Selenomonas flueggei, and Centipeda periodontii. Species not previously reported in studies that used only molecular techniques were identified in the present study.     

Prevalence of Candida albicans and Trichomonas vaginalis in pregnant women in Havana City by an immunologic latex agglutination test

We aimed to estimate the prevalence of Candida albicans and Trichomonas vaginalis in immunocompetent pregnant women living in Havana City, Cuba, with or without symptoms of vaginitis, using a sample of 640 women from 6 Gyneco-obstetrics hospitals, which represents 2.5% of total yearly pregnant women. Diagnosis was made using a new latex agglutination kit (Newvagin C-Kure, La Habana, Cuba). Clinical sensitivity and specificity of this assay were validated against culture method, with 467 and 489 clinical specimens for Candida albicans and Trichomonas vaginalis, respectively. Results showed that the kit clinical sensitivity was 100% for Candida albicans and 86.7% for Trichomonas vaginalis compared with a clinical specificity of 93.3% for Candida albicans and 95.1% for Trichomonas vaginalis by culture. The prevalence of candidiasis was determined to be 42.3% (95% confidence interval [CI] 3.8%); the prevalence of trichomoniasis was 9.84% (95% CI 2.3%). In our sample, 48.7% of the women tested negative with respect to both candidiasis and trichomoniasis. Only 6.41% of the cases yielded inconclusive results. The test has high sensitivity, and our results indicate a relatively high prevalence of both infections. However, a significant difference (P < .001) was also observed in candidiasis and trichomoniasis prevalence among hospitals corresponding to the quantity of women with clinical vaginitis. No difference was observed between diabetics and nondiabetics, probably due to the special care of diabetic pregnant women. We conclude that the method is useful for this kind of vaginitis prevalence study and that candidiasis and trichomoniasis prevalences in pregnant women of Havana are 38.5% to 46.2 % (95% CI) and 7.5% to 12.1% (95% CI), respectively.     

乳杆菌治疗女性阴道炎相关研究进展

健康妇女的阴道中存在黏膜免疫系统及乳杆菌与其它微生物群的动态平衡,对维持其自净及宿主健康起重要作用。乳杆菌在阴道内具有抵抗内源性和外源性病原菌生长繁殖的作用。当这种平衡被打破时,乳杆菌减少而其他微生物大量繁殖,造成阴道炎的发生。对于阴道炎的治疗,临床可以采用微生态治疗——合理使用抗生素和微生态调节剂的应用,从而起到杀菌、修复阴道上皮和恢复阴道微生态环境的作用。     

Equine stomachs harbor an abundant and diverse mucosal microbiota

Little is known about the gastric mucosal microbiota in healthy horses, and its role in gastric disease has not been critically examined. The present study used a combination of 16S rRNA bacterial tag-encoded pyrosequencing (bTEFAP) and fluorescence in situ hybridization (FISH) to characterize the composition and spatial distribution of selected gastric mucosal microbiota of healthy horses. Biopsy specimens of the squamous, glandular, antral, and any ulcerated mucosa were obtained from 6 healthy horses by gastroscopy and from 3 horses immediately postmortem. Pyrosequencing was performed on biopsy specimens from 6 of the horses and yielded 53,920 reads in total, with 631 to 4,345 reads in each region per horse. The microbiome segregated into two distinct clusters comprised of horses that were stabled, fed hay, and sampled at postmortem (cluster 1) and horses that were pastured on grass, fed hay, and biopsied gastroscopically after a 12-h fast (cluster 2). The types of bacteria obtained from different anatomic regions clustered by horse rather than region. The dominant bacteria in cluster 1 were Firmicutes (>83% reads/sample), mainly Streptococcus spp., Lactobacillus spp. and, Sarcina spp. Cluster 2 was more diverse, with predominantly Proteobacteria, Bacteroidetes, and Firmicutes, consisting of Actinobacillus spp. Moraxella spp., Prevotella spp., and Porphyromonas spp. Helicobacter sp. sequences were not identified in any of 53,920 reads. FISH (n = 9) revealed bacteria throughout the stomach in close apposition to the mucosa, with significantly more Streptococcus spp. present in the glandular region of the stomach. The equine stomach harbors an abundant and diverse mucosal microbiota that varies by individual.     

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。