健康妇女和外阴阴道假丝酵母菌病患者阴道乳杆菌对阴道假丝酵母菌抑制作用的实验研究

目的:探讨正常健康和外阴阴道假丝酵母菌病(VVC)妇女阴道分泌物中分离的乳杆菌对假丝酵母菌生长的抑制。方法:健康正常(正常组)和VVC初发(VVC组)妇女各30名,通过定量采集阴道分泌物培养乳杆菌,观察阴道乳杆菌数量及菌种分布情况,并将获得的乳杆菌及VVC患者阴道分泌物中假丝酵母菌分离纯化及鉴定后,采用牛津杯法观察乳杆菌对假丝酵母菌生长的抑制作用,同时比较VVC组和正常组乳杆菌产H2O2能力的差异。结果:①乳杆菌检出率及菌种分布组间无统计学差异(P>0.05);②VVC组阴道乳杆菌数量的平均对数值与正常组相比显著减少(P<0.01);③阴道内乳杆菌BV试剂盒检测为阴性者,正常组显著高于VVC组(P<0.01);④正常组阴道分离的乳杆菌与VVC组比较,对白色和热带假丝酵母菌的生长抑制有统计学差异(P<0.05),对克柔假丝酵母菌的抑制无统计学差异(P>0.05);正常组分离的嗜酸乳杆菌与VVC组的比较,对白色和热带假丝酵母菌的生长抑制均有统计学差异(P<0.01,P<0.05),对克柔假丝酵母菌的生长抑制无统计学差异(P>0.05);但是正常组和VVC组分离的詹氏乳杆菌对上述3种假丝酵母菌的生长抑制均无统计学差异(P>0.05)。结论:①正常妇女与VVC患者阴道内乳杆菌检出率、菌种分布相似,均以嗜酸、詹氏乳杆菌为优势菌种;②VVC患者阴道乳杆菌的数量显著减少;③VVC组产生H2O2的能力下降,可能与VVC发生相关;④正常状态下阴道乳杆菌对假丝酵母菌抑制作用优于VVC感染状态,其中以嗜酸乳杆菌的抑菌作用最为明显。     

乳杆菌功能改变与外阴阴道假丝酵母菌病

目的研究VVC患者阴道乳杆菌菌种构成与功能特点。方法从VVC患者和健康女性阴道分泌物中分离乳杆菌并鉴定。测定乳杆菌抑制白假丝酵母菌的能力。结果对照组女性阴道分泌物分离出64株乳杆菌,81.3%能抑制白假丝酵母菌增殖,VVC组分离出51株乳杆菌,70.6%能抑制白假丝酵母菌增殖。对照组常见菌种为卷曲乳杆菌、加氏乳杆菌和詹氏乳杆菌,VVC组常见菌种为阴道乳杆菌、发酵乳杆菌和德氏乳杆菌。对照组所有患者均能分离到具有抑制白假丝酵母菌功能的乳杆菌,VVC组仅42%的患者能分离到具有抑制白假丝酵母菌作用的乳杆菌。结论 VVC患者阴道乳杆菌菌种构成与健康女性存在差别。VVC组患者阴道乳杆菌抑制白假丝酵母菌能力较对照组弱。     

健康女性阴道乳杆菌抑菌功能的初步研究

目的:研究健康女性阴道乳杆菌对阴道常见致病菌的抑制作用及其机制。方法:选取北京大学第一医院体检中心18～45岁的育龄期健康女性17例,从其阴道分泌物中分离、纯化乳杆菌并进行鉴定。采用spot on lawn和双层牛津杯法筛选出对粪肠球菌、微小消化链球菌、大肠埃希菌和白假丝酵母菌有抑制作用的阴道乳杆菌菌株。测定乳杆菌的生长曲线及代谢物内的乳酸含量。将乳杆菌与白假丝酵母菌共培养,观察乳杆菌菌体和菌液粗提物在抑制白假丝酵母菌的过程中各自所起的作用。结果:17例健康女性阴道分泌物中分离出23株乳杆菌,其中6株具有抑菌作用,分别是加氏乳杆菌、詹氏乳杆菌、阴道乳杆菌、卷曲乳杆菌、发酵乳杆菌和德氏乳杆菌,其中加氏乳杆菌增殖能力较强,詹氏乳杆菌最弱。6株乳杆菌代谢物内乳酸的含量不同,发酵乳杆菌最高为(2.72±0.035)mg/ml,卷曲乳杆菌和德氏乳杆菌较低,分别为(2.14±0.035)mg/ml和(2.13±0.025)mg/ml。加氏乳杆菌、詹氏乳杆菌、阴道乳杆菌的菌液、菌体及菌液粗提物均可以抑制白假丝酵母菌的生长(P≤0.05);发酵乳杆菌和德氏乳杆菌的菌液和菌体可以抑制白假丝酵母菌的生长(P≤0.05);卷曲乳杆菌对白假丝酵母菌则无抑制作用。结论:健康女性阴道乳杆菌的多个菌株均具有抑制大肠埃希菌、粪肠球菌、微小消化链球菌或白假丝酵母菌的功能。詹氏乳杆菌粗提物中细菌素类物质和过氧化氢的联合作用可以抑制白假丝酵母菌,加氏乳杆菌和阴道乳杆菌主要依靠过氧化氢起作用。乳杆菌产生的乳酸没有发挥抑菌作用。     

常用抗假丝酵母菌药物对阴道常见乳杆菌生长影响的研究

目的:探讨治疗外阴阴道假丝酵母菌病(VVC)的药物对阴道常见乳杆菌生长的影响。方法:VVC患者随机分为3组,分别应用咪康唑(咪康唑组)、伊曲康唑(伊曲康唑组)、制霉菌素栓(制霉菌素组)进行治疗,治疗前、后分别进行乳杆菌Nugent评分,治疗后观察患者症状缓解时间、阴道pH值、乳杆菌数量及H2O2阳性乳杆菌的检出率等。结果:患者症状缓解时间、总有效率3组间差异无统计学意义(P>0.05)。3组乳杆菌Nugent评分均较治疗前有所降低,且伊曲康唑组乳杆菌Nugent评分显著高于咪康唑组和制霉菌素组,差异有统计学意义(P<0.05)。咪康唑组、伊曲康唑组、制霉菌素组中H2O2阳性的乳杆菌分别占30.0%、60.0%和25.0%,差异有统计学意义(P<0.05),伊曲康唑组H2O2阳性的乳杆菌的检出率显著高于其余2组,差异有统计学意义(P<0.05)。结论:伊曲康唑治疗VVC疗效确切,对阴道乳杆菌生长的影响较小,有助于维护阴道正常菌群,是一种治疗VVC较理想的抗真菌药物。     

外阴阴道假丝酵母菌病123例临床分析

目的:探讨外阴阴道假丝酵母菌病(VVC)与致病菌种的变迁、药物敏感性的变化和性伴侣感染之间的关系.方法:对妇产科门诊123例VVC患者的阴道分泌物及其性伴侣龟头皮屑进行真菌培养和药敏试验.结果:VVC致病菌株白假丝酵母菌78.9%,光滑假丝酵母菌占18.7%,克柔假丝酵母菌占2.4%.对酮康唑药物敏感率高达75.6%,对特比奈芬耐药率高达75.6%.在随访的102例性伴侣中,有症状者44.1%,镜检阳性率为26.5%,培养阳性率为51.0%,菌种分布与性伴侣一致率为98.0%.结论:非白假丝酵母菌占VVC致病菌比例增加,而假丝酵母菌对唑类药物耐药性有增加趋势,性伴侣的感染可能是VVC复发的重要原因.     

外阴阴道假丝酵母菌病的致病菌研究

目的　了解中国外阴阴道假丝酵母菌病(VVC)的致病菌种,并评价阴道应用硝酸咪康唑 400mg3日疗法与 200mg7日疗法对VVC的疗效。方法　采用多中心大样本开放性研究,对 2003年 1月至 7月全国 6家医院 250例VVC患者进行阴道假丝酵母菌的培养,并比较硝酸咪康唑 400mg3日疗法与 200mg7日疗法的疗效及副反应。结果　 (1)白色假丝酵母菌仍是造成VVC最常见的致病菌,占 88.4%,而其他一些非白色假丝酵母菌依次为光滑假丝酵母菌 ( 5. 2% )、伪热带假丝酵母菌 ( 1.6% )、星型假丝酵母菌 ( 1.2% )、近平滑假丝酵母菌(1.2% )、平滑假丝酵母菌(0.8% )及季也蒙假丝酵母菌 (0.8% )等。(2)停药 10 ~14d时, 400mg组真菌阴转率为 89 9%, 200mg组真菌阴转率为 89 7% (P>0.05)。停药 28~35d时, 400mg组真菌阴转率为 96.1%, 200mg组真菌阴转率为 93 1% (P>0.05)。结论　(1)VVC的最常见致病菌是白色假丝酵母菌,机体的免疫异常及阴道环境的改变是VVC发生的重要原因。(2)硝酸咪康唑 400mg3日疗法与 200mg7日疗法在治疗效果上差异无显著意义。     

外阴阴道假丝酵母菌病的菌种与耐药性研究

目的:对复发性外阴阴道假丝酵母茵病(RVVC)、外阴阴道假丝酵母菌病(VVC)患者阴道分泌物进行培养、菌种分型及药物敏感试验,探讨其治疗策略.方法:用沙氏培养基35℃培养194例患者阴道分泌物,其中RVVC 97例,VVC 97例.用API 32℃假丝酵母茵鉴定板及ATBTM FUNGUS3 药敏板接种,在ATB Expression全自动微生物鉴定仪判读结果.结果:在培养检测的194例阴道分泌物中白假丝酵母菌156例,占80.41%(156/194);RVVC组中73例为白假丝酵母菌,占75.26%(73/97),VVC组中83例为白假丝酵母菌,占85.57%(83/97).RVVC组非白假丝酵母菌比例高,与VVC组比较差异有统计学意义(P＜0.05).RVVC组假丝酵母菌对氟康唑药物敏感率(68.04%)低于VVC组(86.60%)(P＜0.05).结论:VVC、RVVC的主要致病菌仍是白假丝酵母菌,RVVC组非白假丝酵母菌比例高于VVC组.RVVC组假丝酵母菌对氟康唑的敏感性明显低于VVC组.     

外阴阴道假丝酵母菌病临床分离株的基因型和药物敏感性分析

目的:探讨外阴阴道假丝酵母菌病(VVC)患者阴道白假丝酵母菌分离株的基因型别特征及不同基因型别的阴道白假丝酵母菌的药物敏感情况。方法:(1)对211株阴道白假丝酵母菌进行25S rDNA基因分型;(2)采用NCCLS推荐的M27-A微量法测定阴道白假丝酵母菌对5-氟胞嘧啶、咪康唑、伊曲康唑、克霉唑、氟康唑和制菌霉素等6种药物的敏感性。结果:(1)211株阴道白假丝酵母菌分为3种基因型,A型189株(89.6%),B型19株(9.0%),C型3株(1.4%);(2)不含内含子组(A型)白假丝酵母菌对5-氟胞嘧啶、咪康唑、伊曲康唑、克霉唑和氟康唑的最低抑菌浓度值(MIC)高于含内含子组(B、C型)(P0.05);不含内含子组(A型)与含内含子组(B、C型)白假丝酵母菌对5-氟胞嘧啶和咪康唑的敏感率有显著差异(P0.01),前者显著低于后者。结论:不同基因型别的白假丝酵母菌在阴道内的分布具有独特性,A型为主要基因型;VVC白假丝酵母菌基因型别对不同药物的敏感率和MIC有影响。     

细胞溶解性阴道病的诊治进展

乳杆菌是女性阴道正常菌群中的重要成员,可通过产生乳酸、H2O2和其他抑制物质如嗜酸乳菌素,刺激免疫系统等方式来抑制大肠埃希菌、假丝酵母菌、阴道加德纳菌的过度生长,维持阴道微生态平衡。但乳杆菌的过量生长可造成阴道上皮鳞状细胞的破裂,导致一些类似外阴阴道假丝酵母菌病(VVC)的临床症状,即细胞溶解性阴道病(CV)。通过对CV的诊断、临床鉴别、治疗等方面进行文献回顾,提示本病发病率低于VVC、复发性VVC(RVVC)等,临床中易误诊误治。对于长期阴道分泌物异常、实验室真菌检查反复阴性、经验使用治疗真菌感染类药物无效的患者,应积极考虑本病的可能。     

复发性外阴阴道假丝酵母菌病的菌种及药敏分析197例

目的:了解复发性外阴阴道假丝酵母菌病(RVVC)临床分离菌株构成及其对常见抗真菌药物的敏感性。方法:对确诊的197例RVVC患者和466例外阴阴道假丝酵母菌病(VVC)患者的阴道采集标本进行致病假丝酵母菌株培养分离、菌种鉴定和药敏试验。结果:①RVVC与VVC组在假丝酵母菌组成上差异无统计学意义;②RVVC组假丝酵母菌对制菌霉素、咪康唑、酮康唑和氟康唑的敏感性(51.6%、27.5%、30.8%、37.9%)显著低于VVC组(79.7%、47.5%、47.5%、55.1%);对两性霉素B、制菌霉素、咪康唑、益康唑的耐药性(12.1%、10.4%、50.0%、42.9%)显著高于VVC组(3.3%、1.7%、33.1%、22.0%)。结论:RVVC的致病菌株对抗真菌药物的敏感性全面降低而耐药性全面升高,应引起临床的重视。     

In vitro antagonistic effect of Lactobacillus on organisms associated with bacterial vaginosis

OBJECTIVE: To assess antagonistic properties of Lactobacillus strains isolated from the vaginas of healthy women as compared to the most common bacterial agents related to vaginosis. STUDY DESIGN: Antagonistic activity of different Lactobacillus strains isolated from the vaginas of healthy women not treated for infections with an antibiotic for the previous three months was screened using an agar slab method. The activity was tested against test organisms associated with bacterial vaginosis and/or urinary tract infections: Staphylococcus aureus, Enterococcus faecalis, Streptococcus agalactiae, Escherichia coli, Gardnerella vaginalis, Peptostreptococcus anaerobius and Prevotella bivia. RESULTS: Many of the 146 Lactobacillus strains tested exerted apparent antagonistic activities against gram-positive aerobic cocci and gram-negative rods, such as S aureus and E coli, and a marked number of Lactobacillus strains inhibited facultative bacteria, such as Gardnerella vaginalis and the anaerobes P anaerobius and P bivia. Only a few lactobacilli were able to inhibit growth of E faecalis and S agalactiae. Indicator bacteria growth inhibition probably relies upon several different complementary mechanisms. The specific indicator bacteria species determines which mechanism predominates. CONCLUSION: Lactobacillus strains taken from normal vaginal flora demonstrated antagonistic activity against a variety of bacteria related to vaginal and urinary tract infections. The specific occurrence rates of active Lactobacillus strains are different, and this difference is dependent on the indicator bacteria species.     

The in vitro activity of vaginal Lactobacillus with probiotic properties against Candida

Lactobacilli, the predominant vaginal microorganisms in healthy premenopausal women, control other members of the vaginal microflora and thus protect against bacterial vaginosis and urinary tract infections. It has been claimed that some lactobacilli are also protective against Candida vaginitis. Little is known, however, about the mechanisms by which these lactobacilli can control vaginal populations of Candida and prevent vaginitis. To address this question, vaginal Lactobacillus strains with known antagonistic properties against bacteria were tested for their cell surface properties, adhesion to vaginal cell lines in vitro and antagonistic activities against Candida. A small proportion of the lactobacilli tested adhered strongly to cultured vaginal epithelial cells and inhibited growth of Candida albicans but not of C. pseudotropicalis. This anticandidal activity was in some Lactobacillus strains related to hydrogen peroxide (H2O2) production, but catalase treatment did not suppress this activity in other Lactobacillus strains, suggesting alternative mechanism(s). Moreover, tested vaginal Candida strains were resistant to relatively high concentrations of H2O2 that markedly exceeded those produced by even the most active Lactobacillus strains.     

阴道白假丝酵母菌酶活性影响因素的研究

目的:探讨外环境改变对阴道白假丝酵母菌酵母相和菌丝相的酶活性的影响。方法:收集标本50株菌株,用牛奶培养基和卵黄培养基法检测白假丝酵母菌酵母相与菌丝相分泌型酸性蛋白酶(SAP)和磷脂酶(PL)的酶活性;从中随机挑选20株菌株,分别用不同pH值的培养基检测其酵母相和菌丝相酶活性的改变;在不同乳杆菌浓度的环境下检测阴道白假丝酵母菌酵母相和菌丝相酶活性的变化。结果:酵母相和菌丝相磷脂酶的活性(PZ值)分别为0.546±0.132和0.471±0.119;分泌型酸性蛋白酶的活性(PA值)分别为0.605±0.125和0.568±0.086;菌丝相分泌型酸性蛋白酶和磷脂酶活性均显著高于酵母相(P<0.001)。pH=4时,白假丝酵母菌酶活性最低,随着pH值增加,酶活性升高,至pH=7时酶活性降低。未添加乳杆菌组白假丝酵母菌酶活性最高,逐渐增加乳杆菌浓度,酶活性随之降低。结论:菌丝相酶活性均高于酵母相;pH值可影响白假丝酵母菌酶活性的改变,过酸或过碱的环境均可抑制白假丝酵母菌酶活性;乳杆菌对白假丝酵母菌的酶活性有抑制作用,且对菌丝相的抑制作用较酵母相更强。     

Antibacterial properties of human amnion and chorion in vitro

OBJECTIVE: The purpose of the present study was to explore the direct effects of amnion and chorion on bacterial growth in vitro including the antibacterial spectrum. Chorioamniotic membranes were obtained under sterile conditions from 13 healthy women undergoing elective cesarean section at term. Likewise, chorioamniotic membranes were obtained from 10 healthy women with spontaneous vaginal delivery at term. Five strains of Hemolytic streptococci group B (GBS) were tested and one clinical isolate of the following species or bacterial groups: Hemolytic streptococcus group A, Staphylococcus aureus, Staphylococcus saprophyticus, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter calcoaceticus and Lactobacillus species. Bacteriological media included (1) blood-agar medium; (2) a transparent agar medium for submerged cultures; and (3) a nutrient broth medium. RESULTS: An inhibitory effect of fetal membranes against a range of bacteria was found. Consistent results were obtained in experiments with cultures on agar and cultures suspended in agar (membranes from eight women in both studies). In experiments with liquid cultures (seven women) only chorion showed a marginal inhibitory effect. All strains were inhibited, but the most pronounced inhibition was obtained for streptococcus group A, S. aureus and S. saprophyticus by both chorion and amnion. CONCLUSION: This study demonstrated an inhibitory effect of the fetal membranes on a diverse panel of bacteria     

嗜血菌、念珠菌和支原体组合培养基诊断生殖道感染的研究

目的:探讨嗜血菌、念珠菌和支原体组合培养基对妇女生殖道感染的诊断价值。方法:取36例临床标本,分别接种于嗜血菌、念珠菌和支原体组合培养基及传统单一的嗜血菌培养基、科玛嘉显色培养基和支原体培养基上培养,于24 h、48 h观察培养结果。结果:在36例临床标本中,①嗜血菌、念珠菌和支原体组合培养基培养出革兰氏阳性菌9株、革兰氏阴性菌4株;白色念珠菌14株、克柔念珠菌4株、光滑念珠菌3株、其他念珠菌1株;解脲脲原体(ureaplasma urealyticum,Uu)17株,解脲脲原体合并人型支原体(Mycoplasma hominis,Mh)3株。②嗜血菌、念珠菌和支原体组合培养基与传统单一的嗜血菌培养基、科玛嘉显色培养基和支原体培养基对比,对临床标本的检测符合率均高于88%,分离阳性率差异无统计学意义(P>0.05)。结论:嗜血菌、念珠菌和支原体组合培养基可同时培养出细菌、支原体、念珠菌3种微生物,有利于对妇女生殖道感染进行诊断及鉴别诊断,具有指导临床用药的重要意义。     

The inhibitory effect of clindamycin on Lactobacillus in vitro

OBJECTIVE: To evaluate the in vitro effect of varying concentrations of clindamycin on Lactobocillus spp. METHODS: Concentrations of clindamycin ranging from 1.95-20,000 microg/ml were studied for their effect on the growth of six strains of Lactobacillus. RESULTS: Clindamycin concentrations between 1.95-31.25 microg/ml had no statistically significant effect on growth of lactobacilli (p > 0.05). Concentrations 125 and 250 microg/ml had a bacteriostatic effect. The mean minimum inhibitory concentration (MIC) for studied Lactobacillus strains was determined as 1,000 microg/ml. CONCLUSION: High concentrations of clindamycin achieved in the vagina by intravaginal application might be inhibitory for Lactobacillus.     

Phage infection in vaginal lactobacilli: an in vitro study

Objective: During bacterial vaginosis, an unexplained decrease of vaginal lactobacilli occurs. To identify whether these lactobacilli could be infected by phages, we isolated phages from vaginal lactobacilli and analyzed their potential virulence in attacking vaginal lactobacilli in vitro.Methods: Vaginal samples were obtained from 39 reproductive-aged women. The selective Rogosa SL agar was used to isolate lactobacilli, from which phages induced by mitomycin C or released spontaneoulsy were analyzed by the agar spot method.Results: Of 20 samples from women with vaginal infections, 12 did not have lactobacilli. From the remaining 8 infection samples and the 19 samples from healthy women, 37 Lactobacillus strains were isolated, from which 7 temperate phages were identified. Upon analysis, all 7 phages infected vaginal lactobacilli from the same and/or different women in vitro. Two phages, Phikc005 and Phikc007, had a broad host range, infecting 7 of 8 species tested. A control intestinal Lactobacillus phage also lysed several vaginal strains. One vaginal phage, Phikc039, was apparently lytic against vaginal lactobacilli from 7 other women. This phage was characterized as follows: plaque morphology, small and clear; burst size, 300 phages per cell; spontaneous induction rate, 1 per 10(6) cells; DNA, double-stranded and linear, 41 kb; and shape, a hexogonal head and a non-contractile tail.Conclusions: Bacteriophages were isolated from vaginal lactobacilli of some women and were shown in vitro to lyse vaginal Lactobacillus strains from the same and/or different women. It was suggested that vaginal lactobacilli might be suppressed by phages.     

Effect of metronidazole on the growth of vaginal lactobacilli in vitro

OBJECTIVE: To determine whether metronidazole has an adverse effect on the growth of Lactobacillus. METHODS: Hydrogen peroxide- and bacteriocin-producing strains of Lactobacillus were used as test strains. Concentrations of metronidazole used ranged from 128 to 7000 microg/ml. Susceptibility to metronidazole was conducted by the broth microdilution method recommended by the National Committee for Clinical Laboratory Standards. RESULTS: Growth of Lactobacillus was partially inhibited at concentrations between 1000 and 4000 microg/ml (p = 0.014). Concentrations > or = 5000 microg/ml completely inhibited growth of Lactobacillus. Concentrations between 128 and 256 microg/ml stimulated growth of Lactobacillus (p = 0.025 and 0.005, respectively). Concentrations of metronidazole between 64 and 128 microg/ml or > or = 512 microg/ml did not have an inhibitory or a stimulatory effect on the growth of Lactobacillus compared to the control. CONCLUSIONS: High concentration of metronidazole, i.e. between 1000 and 4000 microg/ml, partially inhibited the growth of Lactobacillus. Concentrations > or = 5000 microg/ml completely suppressed the growth of Lactobacillus. Concentrations between > or = 128 and < or = 256 microg/ml stimulated the growth of Lactobacillus. Further investigation to determine the ideal concentration of metronidazole is needed in order to use the antimicrobial agent effectively in the treatment of bacterial vaginosis.