不同条件下小鼠白念珠菌性阴道炎易感性的研究

目的研究不同条件下小鼠阴道粘膜对白念珠菌的易感性。方法建立免疫系统正常小鼠(雌激素化)与免疫抑制小鼠模型,建立初发与复发白念珠菌性阴道炎模型,对比观察不同干预措施下小鼠的一般生长情况、阴道灌洗液涂片细胞学检查结果、阴道组织病理学改变并测定其阴道灌洗液真菌载量。结果免疫抑制组各时点平均CFU水平均高于雌激素化组,且峰值后移,自第4天起两组相比有显著性差异(P<0.05)。雌激素化组、免疫抑制组各时点平均CFU分别与正常对照组相比均显示极显著性差异(P<0.01)。结论处于免疫抑制状态的小鼠阴道粘膜对白念珠菌更具有易感性;再次感染与初次感染不存在易感性差异。     

Local IL-23 Expression in Murine Vaginal Candidiasis and Its Relationship with Infection and Immune Status

IL-12 is a heterodi meric cytokine which con-sist of p40 and p35 and plays a key role in the dif-ferentiation of T helper1 ( Th1)cells[1].Its func-tions in murine vaginal candidiasis have beenrepor-ted[2 ,3].IL-23 is a new member inthe IL-12 familyof heterodi meric cytokines which is composed ofp40 and p19[4]. IL-23 shares some features withIL-12 but they also differ ,functionally and struc-turally . Many of its biological features ,such as itsroleinthe development of murine vaginal candid…     

The Effect of Itraconazole on the Vaginal Candidiasis under Different Immunity Conditions in Mice

To study the effect of itraconazole on the vaginal candidiasis caused by Candida under different immunity conditions, the fungal vaginitis model was established in female ICR mice by in- travaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol or dexamethasone. Mice were divided at random into different groups and then treated with itracona- zole or IFN-γ given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the beginning of the drug treatment. The difference in the effect of itraconazole on the vaginal candidiasis between normal immune system group (group A) and control group (group D) was statistically significant (P<0.01). The difference in the efficacy of itraconazole among immunosuppressed group (group E), immuno-regulated group (group F) and the control group (group G) was statistically significant (P<0.01). But on the 5th, 6th, 7th, 9th, 11th day after the inoculation the average level of colony forming unit (CFU) of groups A, E and F showed no statistically significant difference (P>0.05). It is concluded that the efficacy of itraconazole in the treatment of the vaginal candidiasis under different immunity conditions (groups A, E and F) in mice were all good, but there was no difference in the anti-fungal effect of itraconazole among the three groups.     

Local Expression of Vaginal Th1 and Th2 Cytokines in Murine Vaginal Candidiasis under Different Immunity Conditions

To investigate the expression of vaginal Th1 and Th2 cytokines in rats with experimental vaginal candidiasis under different immune conditions, ICR murine vaginal candidiasis model was established and immno-suppressed murine models of vaginal cadidiasis were established in estrogen-treated mice. Non-estrogen-treated mice were used as controls. The mRNA level of Th1 （IL-2）/Th2 （IL-4, IL-10, TGF-β1） cytokines in murine vaginal tissues was determined by RT-PCR. The cykotine in local tissues was increased to different extent under normal immune condition. IL-2 mRNA was increased during early stage of infection, while IL-10 was increased transiently during late stage of infection. TGF-β1 production was found to be increased persistently. At same time, the expression of IL-2 mRNA was suppressed in immno-suppressed group, and the level of IL-4, IL-10, and TGF-β1 were higher than the normal immunity group to different degree during infection. The high level of IL-2 mRNA during early stage of infection was associated with clearance of mucosal Candidia albicans （C. albicans）, and its expression suppressed leading to decreased clearance of mucosal C. albican in immuno-suppression. The over-expression of IL-4 and IL-10 could significantly enhance the susceptibility to C. albicans infection in mice.     

Expression and Significance of NF-κB, IL-1β and COX-2 in the Murine Model of Estrogen-dependent Experimental Vulvovaginal Candidiasis

Objective To investigate the possible role of estrogen in the pathogenesis of vulvovaginal candidiasis(VVC).Methods Estrogen-dependent experimental murine model of C. albicans vaginal infection was established by injecting subcutaneously with estradiol benzoate and then 5 × 106 stationary-phase C. albicans blastoconidia was inoculated intravaginally to mice (group EI),and other 3 groups were set up: estrogen-treated but not infected (group E) ;estrogen-untreated but infected (group Ⅰ);normal control (group C).The dynamic change of colony-forming unit (CFU) of cervivovaginal lavage fluid was observed. Vaginal tissues at different time points (d 2,d 4,d 7 and d 14) after inoculation of C.albicans were obtained.In situ hybridization staining was used to detect expression of on d 4 and d 7 (P＜0.01).Conclusions In the murine model of estrogen-dependent experimental VVC,estrogen promotes the infection establishment by up-regulating expression of CO X-2 via activating NF-κB signal pathway,and the high expression of COX-2 promoted by the interaction of IL-1β and NF-κB after infection formation was involved in persistence of infection.     

The effect of itraconazole on the vaginal candidiasis under different immunity conditions in mice

Summary To study the effect of itraconazole on the vaginal candidiasis caused by Candida under different immunity conditions, the fungal vaginitis model was established in female ICR mice by intravaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol or dexamethasone. Mice were divided at random into different groups and then treated with itraconazole or IFN-γ given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the beginning of the drug treatment. The difference in the effect of itraconazole on the vaginal candidiasis between normal immune system group (group A) and control group (group D) was statistically significant (P<0.01). The difference in the efficacy of itraconazole among immunosuppressed group (group E), immuno-regulated group (group F) and the control group (group G) was statistically significant (P<0.01). But on the 5th, 6th, 7th, 9th, 11th day after the inoculation the average level of colony forming unit (CFU) of groups A, E and F showed no statistically significant difference (P>0.05). It is concluded that the efficacy of itraconazole in the treatment of the vaginal candidiasis under different immunity conditions (groups A, E and F) in mice were all good, but there was no difference in the anti-fungal effect of itraconazole among the three groups. This project was supported by grants from the Science Research Foundation of Health Department of Hubei Province (No. JXIB048) and the Janssen Research Foundation.     

Study of Candida Albicans Vaginitis Model in Kunming Mice

The model of vaginal candidiasis in Kunming mice was constructed in order to search for the optima construction conditions and provide an economic animal model of Candida albicans (C. albicans) vaginitis. Estrogen benzoate (E2) was given to mice at different concentrations ranging from 0.0 to 0.05 mg/mouse (4 levels) beginning 72 h prior to vaginal inoculation, then mice were in- oculated intravaginally with various concentrations of stationary-phase C. albicans blastoconidia (ATCC90028) (5 levels) in 20 μL of phosphate-buffered saline (PBS) in each E2 level. General state, scores of genital pathology, the hyphae and vaginal fungal burden (CFU) in vaginal lavage fluid, the hydrops rate of uterus and vaginal tissues for pathological section in mice were observed and ob- tained at day 2, 4, 7, 14 and 21 after inoculation. The results showed the infection rate in mice was related to the dosage of E2 and concentration of C. albicans blastoconidia. Additionally there was better cross-effect between the two treated factors. The infection rate was about 80% on the day 4, and could reach 100% on the day 7 until the end of experiment after inoculated intravaginally in groups of E2I3, E2 0.025 mg/mouse injected hypodermically and inoculated intravaginally with 5×104 C. albicans blastoconidia, and large amount of hyphae and blastoconidia could be observe in superfi- cial layer tissue and canal of vaginal by PAS. From the results in our experiment it was concluded that E2I3 was the optima construction condition in kunming mice.     

Comparison of the Effects of Three Different Anti-fungus Drugs on Candida Albicans of Murine Vaginal Mucosa

To compare the therapeutic effects of three different anti-fungal drugs (i.e., terbinafine, fluconazole and intraconazole) in the treatment of experimental vaginitis caused by Candida albicans (C. albicans) in mice, the fungal vaginitis model was established in female ICR mice by intravaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol. Mice were divided at random into different groups and then respectively treated with terbinafine, flucona- zole and intraconazole given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the begin- ning of the drug treatment. The fungal burdens in the vaginal lavage fluids taken at different time points from the mice treated with terbinafine were significantly higher than those taken at corre- sponding time points from mice treated with fluconazole or itraconazole (P<0.01). The fungal bur- dens in the vaginal lavage fluids taken from mice 1 week after the beginning of the treatment with terbinafine remained at a relatively high level. A dramatic drop in the fungal burden was noted in the vaginal lavage fluids taken on the 2nd day of the treatment from mice treated with itraconazole or flu- conazole group and the fungal burden on the 3rd day of the treatment in these mice were at a very low level, suggesting that fluconazole or itraconazole were highly effective for the treatment. How- ever, the difference in the therapeutic effect between the two drugs was not significant (P>0.05). Itraconazole or fluconazole, but not terbinafine, is very effective for the treatment of fungal vaginitis caused by C. albicans in mice.     

脾虚小鼠伴口腔白念珠菌感染唾液中sIgA含量改变

目的:通过测定脾虚小鼠伴口腔白念珠菌感染后唾液中sIgA的含量变化,探讨脾虚小鼠与白念珠菌易感性的关系。方法:采用饮食失节加劳倦过度的复合因素制备脾虚小鼠模型,采用局部接种的方法于正常对照组以及脾虚小鼠建立口腔黏膜白念珠菌感染模型,检测小鼠唾液中活白念珠菌数,应用酶联免疫吸附法(ELISA)测定各组小鼠唾液中sIgA的含量。结果:脾虚伴口腔白念珠菌感染组小鼠的唾液中活白念珠菌数增高,唾液中sIgA含量明显低于其他各组(P<0.01)。结论:小鼠在脾虚状态下对口腔白念珠菌的易感性增强。     

口腔扁平苔藓患者黏膜脱落上皮中人β防御素-2和白色念珠菌的研究

目的 研究口腔扁平苔藓（OLP）患者口腔黏膜脱落上皮中的人β防御素-2（HBD-2）及白色念珠菌的情况。方法 运用RT-PCR和沙堡琼脂培养基细菌培养方法分析10例健康人、70例口腔扁平苔藓患者（35例充血糜烂型、35例斑纹型）口腔黏膜脱落上皮细胞中HBD-2、白色念珠菌的情况。结果 10例健康对照组中HBD-2和白色念珠菌的实验结果均为阴性;70例病例组患者中,HBD-2在充血糜烂型组阳性表达率为45.71%（16/35）,在斑纹型组阳性表达率为22.86%（8/35）,HBD-2在充血糜烂组与在斑纹组中的表达结果比较,差异具有显著性意义（P<0.05）,HBD-2在病例组与在健康对照组的表达结果比较,差异具有显著性意义（P<0.05）;白色念珠菌在充血糜烂型组阳性检测率为37.14%（13/35）,在斑纹型组阳性检测率为14.29%（5/35）,白色念珠菌在充血糜烂组与在斑纹组中的检测结果比较,差异具有统计学意义（P<0.05）;白色念珠菌在病例组中阳性检测率为25.71%（18/70）,与在正常组中的阳性检测结果（0/10）比较,差异具有统计学意义（P<0.05）;病例组中HBD-2与白色念珠菌的相关关系比较,差异无统计学意义（P >0.05）。结论 口腔扁平苔藓患者口腔黏膜脱落上皮中HBD-2的表达情况和白色念珠菌的检测情况与扁平苔藓的病变程度有密切的关系,并随病情的加重所得结果均有增高表现。     

IgY与双歧杆菌防治小鼠白色念珠菌感染的研究

目的 观察特异性抗白色念珠菌鸡蛋黄抗体IgY和双歧杆菌的体内生物学效应，探讨将二者复合应用的可能性。方法 建立无特殊病原菌(Specific pathogen free,SPF)小鼠烧伤后口服白色念珠菌感染模型，应用活菌计数方法检测其回肠膜粘附白色念珠菌量、盲肠内容物白色念珠菌量以及白色念珠菌脏器移位率。结果 烧伤SPF小鼠喂服IgY和双歧杆菌后，均能明显抑制肠道内白色念珠菌生长和白色念珠菌粘附肠上皮细胞；二者复合应用后还能明显降低脏器白色念珠菌移位率。结论 特异性鸡蛋黄抗体IgY具有良好的体内生物学效应且优于单独使用双歧杆菌，二者复合应用效果最佳。     

HIV阳性者唾液sIgA与白色念珠菌黏附能力的关系

目的:研究HIV阳性者易感染白色念珠菌的原因。方法:以HIV阳性者为研究对象,以HIV阴性者为对照,采用放射免疫法测定两者唾液中sIgA含量,用体外黏附方法测定其唾液中白色念珠菌临床株和标准株对颊黏膜上皮细胞的黏附能力,并观察唾液sIgA水平对白色念珠菌黏附颊黏膜上皮细胞能力的影响。结果:HIV阳性者唾液sIgA水平明显低于HIV阴性者唾液sIgA水平(P<0.01);白色念珠菌黏附颊黏膜上皮细胞能力在HIV阳性组唾液中高于在HIV阴性者唾液中的能力(P<0.05);随着HIV阳性者唾液sIgA含量降低,白色念珠菌黏附能力明显增高,呈负相关(r=-0.971)。白色念珠菌临床株和标准株对颊黏膜上皮细胞的黏附能力无显著差异(P>0.05)。结论:HIV阳性者易感染白色念珠菌与唾液sIgA水平密切相关,唾液sIgA水平下降是HIV阳性者易感染白色念珠菌的原因之一,与白色念珠菌菌株来源无关。     

白色念珠菌分泌型酸性蛋白酶和细胞外磷脂酶活性与其毒力的关系

目的： 探讨临床白色念珠菌感染者不同标本中分离的白色念珠菌分泌型酸性蛋白酶和细胞外磷脂酶活性与其毒力的关系。方法： 分别采用牛奶平板和卵黄培养基法检测白色念珠菌感染者不同标本（痰、血液、鹅口疮、伤口、阴道分泌物）中分离的190株白色念珠菌分泌型酸性蛋白酶和细胞外磷脂酶的活力，分别将产酶株的菌丝相和孢子相菌悬液（5×10⁶ CFU•mL^-1）注射小鼠尾静脉，1个月内观察小鼠死亡率及平均存活时间，以平均存活时间评价菌株毒力。结果：190株白色念珠菌分泌型酸性蛋白酶和细胞外磷脂酶阳性检出率分别为83.68%和85.26%。动物实验结果表明，菌丝相白色念珠菌分泌型酸性蛋白酶和细胞外磷脂酶的活力均显著高于孢子相(P<0.01，P＜0.05)；注射菌丝相白色念珠菌的小鼠死亡率高于注射孢子相的小鼠(P<0.05)，且平均存活期短于注射孢子相的小鼠(P<0.01)。结论：分泌型酸性蛋白酶和细胞外磷脂酶是白色念珠菌重要毒力因子，酶活力可直接地反映其毒力和致病性。     

中老年人义齿基托承托区口腔微生物感染研究及影响因素分析

目的 调查中老年人义齿基托承托区可检出口腔微生物感染情况,分析与主要致病菌白假丝酵母菌感染相关的因素。 方法 选择2017年1-9月在杭州师范大学附属医院口腔科就诊的72例患者作为研究对象,分为2组:不戴义齿组(A组)36例和戴义齿组(B组)36例。采集唾液和菌斑培养并初步鉴定菌种,运用单因素和Logistic多因素回归分析可能与白假丝酵母菌感染相关的影响因素。 结果 72例患者标本中可培养微生物206株:厌氧菌176株,其中A组82例,B组94例;白假丝酵母菌30株,其中A组8例,B组22例。B组白假丝酵母菌、黑色素普氏菌及乳杆菌构成比均显著高于A组(P<0.05)。B组变形链球菌构成比显著低于A组(P<0.05)。2组牙龈卟啉单胞菌构成比组间比较差异无统计学意义(P>0.05)。单因素分析证实了吸烟、夜间戴用义齿、义齿卫生和义齿使用年限与白假丝酵母菌检出的相关性,差异均具有统计学意义(P<0.05)。多因素Logistic回归分析提示吸烟、夜间戴用义齿、义齿卫生和义齿使用年限是影响白假丝酵母菌感染的主要因素。 结论 戴用义齿的中老年患者白假丝酵母菌检出例数显著高于不戴义齿中老年患者,其中夜间戴用义齿和义齿卫生是影响白假丝酵母菌感染最重要的因素,更容易引起义齿性口炎。      

卡介苗免疫预防小鼠白色念珠菌感染的实验研究

目的探讨卡介苗（BCG）预先免疫对小鼠白色念珠菌感染的影响。方法采用BCG或灭菌生理盐水皮内注射预先免疫2周,然后由尾静脉注射白色念珠菌进行攻击,观察小鼠死亡率及计算存活小鼠肾组织带菌量。结果BCG免疫组存活时间长于生理盐水免疫组（P〈0.01）;存活小鼠肾组织菌落计数：BCG免疫组远少于省里盐水免疫组（P〈0.01）。结论BCG对小鼠白色念珠菌感染具有很好的保护作用。     

阴道白念珠菌毒力因子表达水平及与耐药的相关性

目的 了解复发性念珠菌性阴道炎（RVVC）患者与阴道白念珠菌定植者的阴道白念珠菌的生物膜、水解酶和菌丝壁蛋白1（HWP1）基因等毒力因子的体外表达水平及其耐药性,初步探究白念珠菌致病性和耐药性与毒力因子之间的关系,为进一步探明白念珠菌引起的RVVC的致病和耐药机制提供资料。方法 前瞻性收集分离来源于成年女性RVVC患者和阴道白念珠菌定植者的100株阴道白念珠菌,采用酵母样真菌药物敏感性试剂盒检测对5种常用抗真菌药物的敏感性,采用结晶紫染色法检测其生物膜形成能力,采用牛血清白蛋白平板、卵黄琼脂平板和三丁酸甘油脂平板检测阴道白念珠菌分泌型天冬氨酸蛋白酶（SAP）、磷脂酶（PL）和脂肪酶（Lip）的活性,采用实时荧光PCR检测HWP1基因的表达。结果 5种常用抗真菌药物中至少1种耐药的白念珠菌RVVC患者来源有13株,阴道定植者来源有10株;RVVC患者来源菌株的生物膜、3种分泌型水解酶和HWP1基因体外表达水平均高于阴道定植者来源菌株（均P<0.05）,两组在生物膜形成强弱、3种水解酶表达高低等定性角度,差异均有统计学意义（均P<0.05）;耐药性菌株生物膜表达水平高于非耐药性菌株外（P<0.05）,对3种分泌型水解酶和HWP1基因体外表达水平上两组间没有差异（均P>0.05）;Pearson相关性分析结果显示,除HWP1基因和生物膜-OD之间呈弱相关性（r=0.211,P=0.039）外,其他各项指标之间没有相关性（均|r|<0.2,P>0.05）。结论 白念珠菌的毒力因子生物膜、SAP、 PL、Lip和HWP1都可能在RVVC的致病过程中发挥了重要作用,HWP1与生物膜的形成呈弱相关性,生物膜的形成与阴道白念珠菌的耐药性产生可能存在关联。     

重组白细胞介素23抗小鼠系统性白假丝酵母菌感染的研究

目的 探讨重组白介素23在抗小鼠系统性白假丝酵母菌感染中的作用.方法 建立环磷酰胺诱导的免疫抑制小鼠系统性白假丝酵母菌感染动物模型,设置对照组(单纯白假丝酵母菌感染组)与处理组(白假丝酵母菌感染前注射重组白介素23).用平皿稀释法检测肾脏、脾脏组织菌落形成单位(CFU)数目;制作肾、脾脏组织病理学标本,评估其病理学分级;并通过酶联免疫吸附试验(ELISA)检测脾脏干扰素γ分泌水平.结果感染后2、3、7 d,处理组肾脏CFU值明显低于对照组(均P＜0.01);处理组脾脏组织CFU值也低于对照组,但差异无统计学意义(P＞0.05).肾脏组织病理学评分处理组低于对照组,处理组较对照组感染程度减轻,差异有统计学意义(P＜0.01);脾脏组织病理学评分处理组也低于对照组,但差异无统计学意义(P＞0.05).同时检测脾脏干扰素γ分泌水平处理组明显高于对照组,差异具有统计学意义(P＜0.01). 结论 重组白介素23在小鼠系统性白假丝酵母菌感染中具有保护作用.     

大蒜素对白色念珠菌感染的免疫抑制小鼠IL-4和IgG产生水平的影响

目的:探讨大蒜素对白色念珠菌感染免疫抑制小鼠体液免疫功能的影响。方法:将实验鼠随机分为预防组、治疗组和感染组。3组小鼠于实验前3 d腹腔一次性注射环磷酰胺200 mg/kg。实验时预防组小鼠灌服大蒜素7 d,其余两组灌服等量生理盐水;第8天各组小鼠尾静脉接种2×107/ml白色念珠菌菌液,同时给予预防组和治疗组分别灌服大蒜素7 d,感染组灌服等量生理盐水7 d;继之,收集3组小鼠脾细胞悬液和血清,测定其IL-4和IgG的表达。结果:与感染组相比,大蒜素可提高预防组和治疗组小鼠IL-4、IgG产生水平(P<0.01),且以预防组作用明显。结论:大蒜素可提高白色念珠菌感染模型鼠体液免疫的功能。     

抗真菌药物治疗外阴阴道念珠菌病敏感性的临床研究

目的探讨外阴阴道念珠菌病(VVC)的临床特征及抗真菌药物治疗VVC的敏感性。方法对我院自2008年1月至2011年8月收治的1000例妇科门诊患者的VVC发病情况进行回顾性分析,观察念珠菌对抗真菌药物的敏感性。结果通过分离1000株念珠菌,白念珠菌为610例(61%),非白念珠菌为390例(39%)。白念珠菌及非白念珠菌对氟康唑和伊曲康唑的敏感度比较差异有统计学意义(P<0.05);对酮康唑和两性霉素B的敏感度比较,差异无统计学意义(P>0.05)。结论白念珠菌是VVC的主要致病菌,非白念珠菌的药物敏感性较白念珠菌低,白念珠菌对抗真菌药物的耐药性不常见。