阴道液中唾液酸酶活性、阴道加德纳菌及其IgA水平的相关性研究

目的探讨阴道液中唾液酸酶活性、阴道加德纳菌及抗阴道加德纳菌的溶血素(anti-Gvh)IgA水平在细菌性阴道病(BV)的相关性。方法对15例健康人和60例临床诊断为BV的患者,取阴道分泌物分别进行唾液酸酶活性和anti-Gvh IgA水平测定及阴道加德纳菌(Gv)分离培养。唾液酸酶活性利用从底物-5溴-4氯-3吲哚基-α-D-N乙酰基神经氨酸(X-Neu5Ac)转化而得到的甲氧基苯酚的纳摩尔数来表示;anti-Gvh IgA通过ELISA法测定。结果BV组Gv活菌数(6.96 log CFU/g)显著高于健康对照组(2.58 log CFU/g)(P<0.01)。BV组anti-GvhIgA(238.0±220.6)显著高于健康对照组(175.0±40.16)(P<0.01)。BV组Gv分离率(86.7%)显著高于健康对照组(33.3%)(P<0.01)。45例临床诊断为BV的患者中,其中26例Gvh IgA(+),19例Gvh IgA(-);Gvh IgA(-)组阴道液中唾液酸酶活性(5.00±1.29)显著高于Gvh IgA(+)组(1.58±1.22)(P<0.01),2组的Gv的分离率和活菌数差异却没有显著性(P>0.05)。结论BV患者阴道内高活性的唾液酸酶与低水平anti-Gvh IgA和黏膜IgA破坏程度具有关联性。     

BV蓝在细菌性阴道炎病诊断中的应用

目的:运用BV蓝快速诊断试剂检测阴道分泌物唾液酸酶活性,观察其在细菌性阴道炎病(BV)诊断中的价值.方法:以无菌棉试子采集符合标准的患者阴道分泌物,分别用BV蓝试剂盒、传统Amsel临床诊断标准进行判断.结果:BV蓝试剂盒在诊断BV中与传统Amsel法符合率为92.6%,其敏感性为96.9%,特异性为88.9%,阳性预测值为88.3%,阴性预测值为97.1%.结论:BV蓝试剂盒检测阴道分泌物唾液酸酶活性是诊断BV的可靠方法,与传统方法比,快速、简便、易于推广.     

BV三联法在细菌性阴道病诊断中的应用与评价

目的探讨BV三联法在诊断细菌性阴道病中的临床价值方法用BV三联法（检测患者阴道分泌物中的过氧化氢浓度、唾液酸苷酶、白细胞脂酶）和Amsel法（检测阴道分泌物并对阴道分泌物进行涂片革兰染色镜检）。结果BV三联法与Amsel法结合涂片革兰染色结果差异无显著性（P〈0．05）。结论BV三联法是一种新型、简便、快速的方法,可用于临床辅助诊断细菌性阴道病。     

650例疑似细菌性阴道病BV患者阴道分泌物检测分析

目的探讨生化检测法在细菌性阴道病（BV）诊断中的应用价值。方法对绍兴市人民医院妇科门诊650例阴道分泌物进行常规及生化检测。结果在650例疑似细菌性阴道病患者中,以“唾液酸苷酶阳性＋过氧化氢阳性”为诊断标准,共检出BV患者483例（占总数的74．31％）;使用Amsel法检出细菌性阴道病患者481例（占总数的74％）。使用生化检测法和Amsel法检出细菌性阴道病的阳性率差异无统计学意义（x2=0．53,P〉O．05）,二者的符合率为94．15％（612／650）,具有很高的一致性。结论生化检测准确、快速,具有较高的临床应用价值。     

BV三联法与Amsel法对检查结果的比较和分析

目的探讨细菌性阴道病联合检测在阴道炎诊断中的临床应用价值。方法对张家港市妇幼保健所妇科就诊的12306例患者进行阴道分泌物常规检查同时进行BV三联联合测定，并与传统的Amsel法作比较。结果H2O2浓度异常〈2μmol／L、唾液酸苷酶阳性和白细胞酯酶阳性率分别61．1％、19．7％和33．3％，诊断细菌性阴道炎的灵敏度分别为94．2％、89．4％和83．6％，特异性分别为64．1％、95．8％和98．9％，两方法差异有统计学意义（P〈0．01）。结论BV三联检测对传统的Amsel法有互补和替代作用，可作为妇产科白带检查的常规筛查项目。     

五联检检测3988例阴道分泌物结果分析

目的 探讨过氧化氢浓度、唾液酸酶、白细胞酯酶、脯氨酸氨基肽酶和乙酰氨基葡萄糖苷酶检测对阴道微生态的改变所致妇科疾病的诊断意义.方法 对妇科门诊3 988例阴道分泌物进行五联检检测及Amsel法的检测.结果 五联检法检测3 988例阴道分泌物的结果阳性率:细菌性阴道病(BV)为30.1%,念珠菌为40.1%,滴虫为21....     

阴道加德纳菌检出率及唾液酸酶A基因携带与细菌性阴道病的关系

目的探究阴道加德纳菌(Gardnerella vaginalis)检出率及唾液酸酶A基因携带与细菌性阴道病(BV)的关系。方法选择2017年1月至2019年8月确诊的BV患者82例作为BV组,并随机选择同时期健康女性82例作为健康组,比较2组人群G. vaginalis检出率和唾液酸酶A基因携带情况,相关统计学资料分析其对BV发生的影响。结果BV组人群G. vaginalis阳性检出率高于健康组(χ²=11.511,P<0.05)。BV组人群共检出G. vaginalis 1、2、3、4、5、6和7型,其中2型占比最高;BV组人群G. vaginalis 2、3、4型占比率高于健康组(χ²=4.148,17.009,9.973,均P<0.05)。BV组人群唾液酸酶A基因携带率高于健康组(χ²=39.234,P<0.05)。较健康组人群,BV组PCR DGGE宽度更窄,肠道菌群条带数少(t=9.217,P<0.05)。结论G. vaginalis检出率和唾液酸酶A基因携带情况与BV发生相关,有待成为相关生物学治疗靶点。     

BV三项在阴道分泌物检测中的应用及临床结果分析

目的探讨阴道分泌物BV三项检测在妇科患者诊断细菌性阴道病中的临床价值及BV阳性患者与妇女盆腔炎症的相互关系。方法对520例阴道分泌物进行BV三项检测（唾液酸苷酶、过氧化氢酶、白细胞酯酶）和显微镜检测,并对其中正常阴道组、滴虫性阴道炎和霉菌性阴道炎与BV进行对照比较。结果根据临床表现和BV三项检测,BV的发病率高达50．96％,滴虫性阴道炎占8．65％,霉菌性阴道炎占17．10％,BV的盆腔炎症明显高于各类阴道病组。结论BV在妇科门诊的发病率为最高,并与妇科盆腔炎症呈正相关性,在妇科门诊进行BV常规筛选是必需的。     

妊娠中晚期阴道微生态失调及乳酸菌胶囊干预对不良妊娠结局的作用

目的 观察妊娠中晚期妇女阴道微生态状况,探讨应用乳杆菌活菌胶囊纠正阴道微生态失调对不良妊娠结局的预防价值。方法 选择孕13~36周单胎妊娠期妇女560例,取其阴道分泌物,经革兰染色后油镜下观察,进行阴道微生态（阴道菌群的密集度、多样性、优势菌、炎症反应等）状况评价,检测阴道分泌物成分、阴道病原菌类型。对阴道微生态失调孕妇,根据是否接受乳杆菌活菌胶囊治疗分为治疗组和对照组,治疗组给予乳杆菌活菌胶囊,对照组不采用药物干预。追踪随访所有孕妇的妊娠情况,比较阴道微生态正常组、微生态失调治疗组及微生态失调对照组的不良妊娠结局。结果 560例研究对象中,阴道微生态正常 335 例（59.82%）,微生态失调225例（40.18%）。225例微生态失调孕妇中,细菌性阴道病（bacterial vaginosis,BV）32例（14.22%）,阴道假丝酵母菌病（vulvovaginal candidiasis,VVC）56例（24.89%）,滴虫性阴道炎（triehomonal vaginitis,TV）11例（4.89%）,BV和VVC混合感染4例（1.78%）,BV和TV混合感染3例（1.33%）,菌群增殖过度75例（33.33%）,菌群抑制44例（19.56%）。微生态失调组pH值>4.5、过氧化氢、白细胞酯酶、唾液酸苷酶、脯氨酸氨基肽酶、乙酰氨基葡萄糖苷酶阳性比例均明显高于微生态正常组（χ2=55.59~340.06,Ps0.05）。结论 妊娠中晚期容易导致阴道微生态失调,造成不良妊娠结局,乳杆菌活菌胶囊纠正阴道微生态失调对于改善不良妊娠结局有较好的预防作用。     

乳杆菌活菌制剂治疗细菌性阴道病疗效观察

细菌性阴道病（Bacteral Vaginosis ,BV）是妇产科常见病,多发病,严重困扰着妇女的身体健康。以往多以抗菌消炎治疗为主,疗效不甚满意,该文以深圳市万泽医药有限公司提供的乳杆菌活菌制剂（定君生）治疗40例细菌性阴道病及用甲硝唑栓治疗40例细菌性阴道病,观察对比评价其疗效。     

Genetic and biochemical diversity of Gardnerella vaginalis strains isolated from women with bacterial vaginosis

Gardnerella vaginalis is considered a substantial player in the progression of bacterial vaginosis (BV). We analysed 17 G. vaginalis strains isolated from the genital tract of women diagnosed with BV to establish a potential link between genotypes/biotypes and the expression of virulence factors, vaginolysin (VLY) and sialidase, which are assumed to play a substantial role in the pathogenesis of BV. Amplified ribosomal DNA restriction analysis revealed two G. vaginalis genotypes. Gardnerella vaginalis isolates of genotype 2 appeared more complex than genotype 1 and were subdivided into three subtypes. Biochemical typing allowed us to distinguish four different biotypes. A great diversity of the level of VLY production among the isolates of G. vaginalis may be related to a different cytotoxicity level of the strains. We did not find any correlation between VLY production level and G. vaginalis genotype/biotype. In contrast, a link between G. vaginalis genotype and sialidase production was established. Our findings on the diversity of VLY expression level in different clinical isolates and linking sialidase activity with the genotype of G. vaginalis could help to evaluate the pathogenic potential of different G. vaginalis strains.     

Response of Gardnerella vaginalis biofilm to 5 days of moxifloxacin treatment

Polymicrobial communities are often recalcitrant to antibiotics. We tested whether the polymicrobial Gardnerella vaginalis biofilm can be eradicated with moxifloxacin. Twenty women with bacterial vaginosis were treated with 400 mg moxifloxacin for 5 days. The changes in the occurrence and proportions of Gardnerella, Atopobium and Lactobacillus spp. were assessed using FISH. The bacterial biofilm was investigated using desquamated epithelial cells of spontaneously voided urine and sections of vaginal biopsies. Fifteen of 20 women showed a significant and sustained clinical response to moxifloxacin according to Amsel and Nugent criteria. The concentrations of adherent bacteria decreased significantly. The incidence and proportion of Atopobium declined sustainably. The proportions of Lactobacillus in the biofilm mass increased following therapy. Initially, Gardnerella was the main component of the polymicrobial biofilm. Following treatment, Gardnerella was not accessible to FISH in the urine and vaginal samples of 75% of all women. Ten to 12 weeks after the end of therapy, Gardnerella biofilm was cumulatively present in 40%. This was not due to newly acquired disease, but due to reactivation of the persisting, but biochemically inactive biofilm. Despite clear clinical efficacy, and initially definite suppression of the biofilm, moxifloxacin was, similar to metronidazole, not able to eradicate the Gardnerella vaginalis biofilm in all patients.     

Fluorescent cytochemical detection of sialidase activity using 5-bromo-4-chloroindol-3-yl-α-D-N-acetylneuraminic acid as the substrate

A novel fluorescent cytochemical method for sialidase activity was developed using 5-bromo-4-chloroindol-3-yl-alpha- D- N-acetylneuraminic acid (X-Neu5Ac) as the substrate. Intact nuclei were isolated from porcine liver and incubated at 37 degrees C for 3 h with 1 mM X-Neu5Ac at pH 4.8. The nuclei were stained with blue color that was derived from the oxidized compound of the reaction product X (5-bromo-4-chloro-3-hydroxyindole). A specific sialidase inhibitor, 2,3-dehydro-2-deoxy- N-acetylneuraminic acid, suppressed the staining in a dose-dependent manner. Despite the specificity of the cytochemical reaction, the staining was too weak to analyze the staining distribution and pattern of individual nuclei. To attain more sensitive detection of sialidase activity, the nuclei were incubated with X-Neu5Ac in the presence of Fast Red Violet LB. Individual nuclei of porcine liver were clearly stained with fluorescence that was produced by the conjugated compound of product X with Fast Red Violet LB. This fluorescent cytochemical method was also employed successfully for detection of sialidase activity of intact GOTO neuroblastoma cells in culture. The present method should provide a useful tool for investigating the localization and stage-specific expression of sialidase activity in tissues and cells.     

健康妇女及3例细菌性阴道病患者阴道菌群的非培养分析

根据Amsel标准及Nugent标准,确诊筛选健康妇女及细菌性阴道病(bacterial vaginosis,BV)患者各3例.提取其阴道分泌物样本的总DNA,构建16S rRNA基因克隆文库,并对阳性克隆进行ARDRA和测序分析.结果表明,健康妇女样本的基因文库中,分别以卷曲乳酸杆菌(L.crispatus)和惰性乳酸杆菌(L. iners)的克隆子占较大比例,另外存在少量的阴道乳酸杆菌(L.vaginalis)或詹氏乳酸杆菌(L.jensenii)的克隆子.BV患者样本的基因文库中,克隆子所代表的菌种类型明显增多,但均以阴道加德纳氏菌(Gardnerella vaginalis)和阴道阿托波氏菌(Atopobium vaginae)的克隆子占较大比例,且无乳酸杆菌克隆子.说明健康妇女阴道菌群的种类单一,以乳酸杆菌占优势,L. iners为优势菌种之一;BV患者阴道菌群的种类复杂多样,但均以Gardnerella vaginalis及Atopobium vaginae共同占优势.     

健康妇女及3例细菌性阴道病患者阴道菌群的非培养分析

根据Amsel标准及Nugent标准, 确诊筛选健康妇女及细菌性阴道病(bacterial vaginosis, BV)患者各3例。提取其阴道分泌物样本的总DNA, 构建16S rRNA基因克隆文库, 并对阳性克隆进行ARDRA和测序分析。结果表明, 健康妇女样本的基因文库中, 分别以卷曲乳酸杆菌(L. crispatus)和惰性乳酸杆菌(L. iners)的克隆子占较大比例, 另外存在少量的阴道乳酸杆菌(L. vaginalis)或詹氏乳酸杆菌(L. jensenii)的克隆子。BV患者样本的基因文库中, 克隆子所代表的菌种类型明显增多, 但均以阴道加德纳氏菌(Gardnerella vaginalis)和阴道阿托波氏菌(Atopobium vaginae)的克隆子占较大比例, 且无乳酸杆菌克隆子。说明健康妇女阴道菌群的种类单一, 以乳酸杆菌占优势, L. iners为优势菌种之一; BV患者阴道菌群的种类复杂多样, 但均以Gardnerella vaginalis及Atopobium vaginae共同占优势。     

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

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。     

Detection of bacterial vaginosis-related organisms by real-time PCR for Lactobacilli,Gardnerella vaginalis and Mycoplasma hominis

The aim of this study was to determine the feasibility of detecting bacterial vaginosis (BV)-related organisms in stored genital tract specimens using real-time PCR. Frozen cervicovaginal lavage (CVL) samples from 21 women were analyzed by real-time PCR for the numbers of Mycoplasma hominis, Gardnerella vaginalis and lactobacilli. Lactobacilli organisms were detected in all CVL samples, G. vaginalis was detected in all but one sample, while M. hominis was detected in only six samples. Using the Amsel criteria to define BV, the samples from women with BV had significantly higher numbers of G. vaginalis organisms than samples from women without BV (P=0.004). In contrast, the number of lactobacilli organisms in BV samples was significantly lower (P=0.013). The number of M. hominis organisms was not significantly different between BV-positive and BV-negative samples. A striking relationship was observed where most of the samples contained high numbers of either lactobacilli or G. vaginalis but not both. These results show that it is possible to determine the presence of BV-related organisms in stored genital tract samples by PCR, suggesting that this could be developed into an objective method that could be useful for certain applications.     

Evidence for Gardnerella vaginalis uptake and internalization by squamous vaginal epithelial cells: implications for the pathogenesis of bacterial vaginosis

Bacterial vaginosis (BV), a common condition seen in premenopausal women, is associated with preterm labor, pelvic inflammatory disease, and delivery of low birth weight infants. Gardnerella vaginalis is the predominant bacterial species associated with BV, although its exact role in the pathology of BV is unknown. Using immunofluorescence, confocal and transmission electron microscopy, we found that VK2 vaginal epithelial cells take up G. vaginalis after exposure to the bacteria. Confocal microscopy also indicated the presence of internalized G. vaginalis within vaginal epithelial cells obtained from a subject with BV. Using VK2 cells and (35)S labeled bacteria in an invasion assay, we found that a 1 h uptake of G. vaginalis was 21.8-fold higher than heat-killed G. vaginalis, 84-fold compared to Lactobacillus acidophilus and 6.6-fold compared to Lactobacillus crispatus. Internalization was inhibited by pre-exposure of cells to cytochalasin-D. In addition, the cytoskeletal protein vimentin was upregulated in VK2 cells exposed to G. vaginalis, but there was no change in actin cytoskeletal polymerization/rearrangements or vimentin subcellular relocalization post exposure. Cytoskeletal protein modifications could represent a potential mechanism for G. vaginalis mediated internalization by vaginal epithelial cells. Finally, understanding vaginal bacteria/host interactions will allow us to better understand the underlying mechanisms of BV pathogenesis.     

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。     

乳杆菌泡腾片治疗细菌性阴道病

目的观察10例BV患者采用了信谊制药总厂生产的阴道嗜酸乳杆菌泡腾片1个疗程(7 d)后阴道菌群的变化。方法采用7种选择或非选择培养基对阴道分泌物的细菌总数、乳杆菌等7个指标进行检测。结果治疗后阴道中乳杆菌显著增多,厌氧革兰阴性杆菌显著减少(两者P<0.001)。细菌总数、肠杆菌、酵母菌和阴道加德纳菌均减少(P<0.01)。结论信谊阴道嗜酸乳杆菌泡腾片具有较好的调整阴道微生态平衡的作用。