念珠菌性外阴阴道炎患者阴道分泌物中白念珠菌分泌型天冬氨酸蛋白酶表达

目的 探讨白念珠菌分泌型天冬氨酸蛋白酶(SAP)家族在白念珠菌无症状性阴道寄居向症状性阴道炎发展过程中的表达.方法 分别从10例无症状携带者、14例念珠菌性外阴阴道炎患者和10例复发性念珠菌性外阴阴道炎患者的阴道分泌物标本中直接提取RNA,逆转录成cDNA后进行RT-PCR.结果 所有无症状携带者和患者均表现出对SAP2和SAP4-6的阳性扩增.10例无症状携带者中无一个标本表现出对SAP1和SAP3的阳性扩增,而念珠菌性外阴阴道炎患者和复发性念珠菌性外阴阴道炎患者阴道分泌物标本提取的RNA可见9个SAP基因的不同表达,并且均不同程度地表现出对SAP1和SAP3的阳性扩增.结论 念珠菌性外阴阴道炎的发病可能与SAP基因的不同表达有关.     

白念珠菌分泌型天冬氨酸蛋白酶基因在急性念珠菌性外阴阴道炎中的表达

目的 了解白念珠菌分泌型天冬氨酸蛋白酶(Sap)基因在人类阴道白念珠菌感染时的表达情况。方法 收集9例阴道念珠菌检测阴性及20例阴道念珠菌感染者的阴道分泌物,通过用特异引物系列进行RT-PCR反应来评价Sap1~Sap6在人体阴道环境中的表达情况。结果 Sap2和Sap5在阴道白念珠菌感染者中是最主要的表达基因;Sap3和Sap4在所有检查者中均被检测到;所有6种Sap基因在某些阴道念珠菌病患者中同时被检出。结论 Sap基因可能与阴道念珠菌病的发病机制有关。     

微卫星基因分型法分析生殖器白念珠菌感染的传播特点

目的 建立适用于株间鉴别的白念珠菌快速微卫星基因分型方法,探索生殖器白念珠菌感染特点.方法 采集39例女性和27例男性生殖器念珠菌病患者生殖器、肛管和口腔的白念珠菌分离株.以三色荧光标记引物,PCR扩增白念珠菌保守基因CDC3、EF3和HIS3微卫星序列,经聚丙烯酰胺凝胶电泳,行微卫星多态性基因分型.结果 三基因联合分析显示,男女生殖器白念珠菌感染者共获18种基因型,主要致病菌株基因型为116:124、122:131、160:200,占感染者的50%以上.3种共有基因型占71%.女性患者中肛管部位菌株基因型与生殖器部位完全一致的占80%,男性则仅占3.8%.女性患者口腔部位的菌株基因型与生殖器部位完全一致的仅占2.7%,男性未见口腔和生殖器一致的菌株基因型.71%的夫妻共患者间生殖器白念珠菌基因型完全相同,其中80%的致病菌株基因型为两性皆感染的主要致病性白念珠菌.结论 改良的白念珠菌微卫星多态性基因分型法能够特异、准确、稳定和快速地进行菌株间鉴别.生殖器白念珠菌感染存在优势基因型.     

Oligo芯片技术分析阴道念珠菌病患者阴道分泌物中致病相关因子的表达谱

目的 借助Oligo基因芯片的优势,整体地分析探讨阴道念珠菌病患者阴道分泌物中各致病相关因子的表达。方法 分别提取10例阴道念珠菌病患者及3例无症状带菌者阴道分泌物标本的RNA,将其与疾病相关因子Oligo表达谱芯片杂交,筛选出表达改变超过2倍（比值≥2或≤0.5）的因子,并绘制成差异基因表达谱进行分析。结果 与正常人对照组相比,在患者组中表达上调的基因有44个,其中,MIP-1α、NF-κB、TNF-α、IFN-γ、TLR4、HWP1、SAP2、SAP5、LIP4、EFG1、CPH1在80%以上的标本中表达上调（平均比值为4.013）;表达下调的基因有17个,其中,LIP6、WH11在80%以上的标本中表达下调（平均比值为0.326）。分析表达差异明显因子的生物信息功能,MIP-1α、NF-κB、TNF-α、IFN-γ、TLR4与机体天然免疫相关,HWP1与菌丝黏附及形成相关,SAP2、SAP5、LIP4、LIP6与菌株胞外水解酶相关,EFG1、CPH1、WH11与菌株表型转换有关。结论 宿主适应性免疫功能受限以及致病菌株毒力增强均参与阴道念珠菌病的发病机制,TLR4在该病局部宿主免疫机制中可能起一定作用。     

抗白念珠菌芽管单抗MAb03.2C1-C2特异性及临床初步应用研究

目的分析抗白念珠菌芽管胞壁外膜抗原单克隆抗体MAb03.2C1-C2的特异性及其应用于实验室检测的可行性。方法用临床分离白念珠菌、致病非白念的念珠菌的孢子、菌丝及常见的酵母菌、细菌做抗原包被,用间接免疫荧光(IIF)方法,对单抗特异性进行分析。取临床口腔念珠菌病患者的真菌涂片菌丝阳性的标本,用IIF方法检测。结果MAb03.2C1-C2仅与白念珠菌芽管或菌丝特异性地结合,与白念珠菌的孢子不发生结合。13种非白念的念珠菌孢子和菌丝、新型隐球菌、大肠杆菌、金黄色葡萄球菌、绿脓杆菌均不能与该单抗相结合。59例口腔念珠菌病真菌涂片IIF试验阳性的标本最终鉴定为白念珠菌(100%)。结论MAb03.2C1-C2对白念珠菌芽管有高度的特异性,并可用于口腔念珠菌病真菌涂片中白念珠菌的早期诊断。     

白念珠菌保护性单抗的研究

目的 探讨抗白念珠菌单克隆抗体在系统性念珠菌感染动物中的保护作用。方法 制备抗白念珠菌单抗,观察单抗对系统性念珠菌感染小鼠存活时间,组织病理改变以及组织中菌落形成单位的影响。结果 制备出3株抗白念珠菌胞壁外膜抗原单克隆抗体-1B5、3E8、4C7;1B5、3E8两株单抗能显著延长致死量白念珠菌感染小鼠存活时间,减少感染小鼠主要脏器组织中白念珠菌菌落形成单位,减轻组织病理改变;1B5单抗能识别白念珠菌胞壁外膜上相对分子质量约为32000抗原;并在体外能抑制白念珠菌孢子对人颊粘膜上皮细胞,胎儿脐静脉内皮细胞的粘附。结论 1B5、3E8是具有保护作用的抗白念珠孢子对人颊粘膜上皮细胞,胎儿脐静脉内皮细胞的粘附。结论 1B5、3E8是具有保护作用的抗白念珠菌单抗;其中1B5是抗白念珠菌胞壁外膜上相对分子质量为32000的抗原的单抗;1B5单抗可通过抑制白念珠菌对上皮细胞,内皮细胞的粘附,降低该菌的侵袭力。     

CHRONIC HYPERPLASTTC CANDIDIASIS—CANDIDAL LEUKOPLAKIA

SUMMARY. Twelve cases of chronic hyperplastic candidiasis of the oral mucosa are described.
Clinically the lesions could not be reliably differentiated from "leukoplakia" due to other causes. All but 2 of the patients were otherwise healthy.
The histological appearances were distinctive: there was invasion by hyphae of C. albicans of the superficial epithelium of an acanthotic and parakeratotic plaque, associated with a characteristic intraepithelial inflammatory reaction.
In 9 cases the antibody titres to C. albicans in serum and saliva supported a diagnosis of candidiasis.
The response of the lesions to oral Nystatin was good in 3 cases and incomplete in 4.
The evidence that C. albicans plays an essential causative rather than a secondary role is critically discussed.
Recognition of chronic hyperplastic candidiasis from other "leukoplakias" is of practical importance since this condition is potentially reversible by medical treatment.     

Oligo基因芯片和小鼠模型研究外阴阴道念珠菌病的发病机制

目的 探讨外阴阴道念珠菌病的发病机制。方法 将来自感染组、阴性对照组及空白对照组小鼠模型的标本分别与自制的疾病相关因子基因表达谱芯片杂交,将感染组与对照组中各疾病相关因子信号值进行比较,筛选出表达升高或降低2倍的因子,并绘制成差异基因表达谱进行分析。结果 与空白对照组相比在感染组中可以出现表达上调的基因有39个,可以出现表达下调的基因有4个。在宿主免疫方面,炎性趋化因子普遍表达增高,适应性免疫调节因子IL-1、IL-4、IL-6、IL-10、IL-12、TNF-α、NF-κB以及TGF-β均有不同程度增强,全部标本天然免疫成分TLR4的表达增高,TLR2仅表达增高于1/3的标本。在致病菌株毒力方面,菌丝生成调控因子EFG1、分泌型天冬氨酸蛋白酶（SAP）2、4、5、6、10、脂酶（LIP）2、LIP4和菌丝胞壁蛋白（HWP）1表达显著增强。其中,单核细胞趋化蛋白（MCP）-1、巨噬细胞炎性蛋白（MIP）-1α和MIP-2、IL-1、Toll样受体（TLR）4、LIP4、HWP1在全部感染组标本中显著增高,这些因子涉及白念珠菌胞外水解酶、菌丝形成、表型转换及宿主天然免疫过程。结论 适应性免疫的功能不足以及致病菌株的毒力增强均参与小鼠阴道念珠菌病的发病机制,TLR4在该病局部宿主免疫机制中可能起较为重要的作用。     

白念珠菌分泌型天冬氨酸蛋白酶研究进展

念珠菌病是一种常见的真菌病,大多数是由念珠菌属中最常见的条件致病真菌白念珠菌引起。分泌型天冬氨酸蛋白酶是与白念珠菌毒力有关的重要因素之一,对其生化特性、编码基因的表达及调控,与宿主的作用加以综述,从而帮助我们更好的了解白念珠菌的致病机制并寻找治疗念珠菌病的新途径。     

念珠菌性阴道炎模型小鼠阴道黏膜中CD11c+树突状细胞的变化

目的:研究白念珠菌感染后,小鼠阴道黏膜树突状细胞(DC)的变化。方法:8~10周龄ICR系小鼠,建立激素依赖型小鼠念珠菌性阴道炎模型,于感染后第2、4、7、14、21d取材,进行阴道灌洗液真菌载量分析及组织病理观察并用免疫组化方法检测小鼠阴道黏膜DC的变化。结果:实验组小鼠阴道灌洗液真菌载量显示,自接种后第2天起即有高水平的CFU计数,并持续到观察期末;阴道组织病理学检查可见大量菌丝附着黏膜表面和黏膜组织内,并有炎性细胞浸润;白念珠菌感染后小鼠阴道黏膜内CD11c DC平均光密度值明显高于对照组(P<0.05)。结论:白念珠菌感染后小鼠阴道黏膜CD11c DC的表达增高,CD11c DC可能参与宿主抗念珠菌感染的免疫机制。     

Vulvovaginal Candida infection: current perspectives

Candida Vulvovaginitis remains the subject of considerable research attention. Yeast species other than Candida albicans are being incriminated with increasing frequency as causes of vaginitis in some recent publications, while analysis of C. albicans strain types by DNA fingerprinting has provided preliminary evidence that the fungus may be able to make minor adaptations of its genotype that facilitate vaginal colonization. The current perception of C. albicans as a potentially virulent microbe relates several molecular factors to invasive processes. These include secretion of hydrolytic enzymes, in particular a proteolytic enzyme, and an inherent tendency to rapid phenotypic switching. Molecular biological experimentation with the genes determining secretion of enzymes should be able to demonstrate definitively the role played by such proteins in the pathogenetic process.     

纤维粘连蛋白在介导白念珠菌与宿主细胞粘附中的作用

为了弄清纤维粘连蛋白（Fn）在介导白念珠菌（简称白念）与宿主细胞粘附中的作用,把人颊粘膜上皮细胞与白念孢子共同振荡孵育2小时,显微镜下测定上皮细胞表面粘附的白念数目,粘附20个以上的上皮细胞记为+,共检测100个,计算百分比为44．75±6．82,再将纤维粘连蛋白（Fn）与上皮细胞和白念共同孵育,具体方法同上,百分比为26.25±5.06,P＜0.01。验证了Fn可能介导上皮细胞与白念的粘附。从受体水平探讨了白念与宿主细胞的粘附机制,为防治白念感染提供了新的途径。     

Antibody monstration in blastomyces infections]

The evidence of humoral antibodies against yeasts can be a valuable help in the diagnosis of such infections. It is however necessary to use several sensitive methods, which complete one another in the detection of the different immunoglobulins. In the interpretation of the results the titer dynamic is decisive, especially in those patients whose antibody answer is suppressed by their disease and/or their therapeutic treatment. In most of the yeast infections Candida albicans can be used as antigen for the detection of antibodies because there is a strong relationship between the antigens of most of the yeasts important in human infections. C. krusei, C. pseudotropicalis and C. guilliermondii do not display enough common antigens to make a diagnosis against these yeasts possible with C. albicans. In that case homologous antigens have to be used. Cross reactions with other microorganisms are less frequent than usually supposed. Therefore so called false positive Candida titers have to be examined very carefully by clinical signs and culture methods.     

Pathomechanisms in Candida albicans infections

The adherence of the pathogen to the host cell is the first and decisive step in Candida albicans infections of the human organism. The mechanisms involved are mostly examined in human epithelial cells which are newly exfoliated and which therefore have a limited life expectancy. In this case, the glycoproteins on the cell surface of Candida albicans--mainly the mannan--are of central importance. They are the main component of the fungus to adhere to the host cell as well as to bind and activate the serum complement. Finally, the role of mannan in the interaction with host defence mechanisms is stressed.     

Vaginal yeast colonisation, prevalence of vaginitis, and associated local immunity in adolescents

OBJECTIVES: To evaluate point prevalence vaginal yeast colonisation and symptomatic vaginitis in middle adolescents and to identify relation of these yeast conditions with reproductive hormones, sexual activity, sexual behaviours, and associated local immunity. METHODS: Middle adolescent females (n = 153) were evaluated for sexually transmitted infections (STIs), asymptomatic yeast colonisation, and symptomatic vulvovaginal candidiasis (VVC) by standard criteria. Also evaluated were local parameters, including vaginal associated cytokines, chemokines, and antibodies, vaginal epithelial cell antifungal activity, and Candida specific peripheral blood lymphocyte responses. Correlations between yeast colonisation/vaginitis and local immunomodulators, reproductive hormones, douching, sexual activity, condom use, and STIs were identified. RESULTS: Rates of point prevalence asymptomatic yeast colonisation (22%) were similar to adults and similarly dominated by Candida albicans, but with uncharacteristically high vaginal yeast burden. In contrast with the high rate of STIs (18%), incidence of symptomatic VVC was low (<2%). Immunological properties included high rates of Candida specific systemic immune sensitisation, a Th2 type vaginal cytokine profile, total and Candida specific vaginal antibodies dominated by IgA, and moderate vaginal epithelial cell anti-Candida activity. Endogenous reproductive hormones were in low concentration. Sexual activity positively correlated with vaginal yeast colonisation, whereas vaginal cytokines (Th1, Th2, proinflammatory), chemokines, antibodies, contraception, douching, or condom use did not. CONCLUSION: Asymptomatic vaginal yeast colonisation in adolescents is distinct in some ways with adults, and positively correlates with sexual activity, but not with local immunomodulators or sexual behaviours. Despite several factors predictive for VVC, symptomatic VVC was low compared to STIs.     

小鼠口腔黏膜白念珠菌感染模型局部ToII样受体2和4 mRNA表达的研究

目的：研究小鼠口腔黏膜白念珠菌感染后局部组织Toll样受体(TLR)2、4mRNA表达的变化规律,探讨其在口腔黏膜抗白念珠菌感染早期免疫中的作用。方法：采用局部接种的方法建立小鼠口腔黏膜白念珠菌感染模型。在不同时间取口腔组织．用半定量逆转录(RT)-PCR检测口腔组织TLR2、TLR4 mRNA表达水平。结果：接种前口腔组织内有微量的TLR2、TLR4 mRNA表达．接种后6h组织中TLR2、TLR4 mRNA表达开始增加,12～24h表达水平达峰值。TLR4 mRNA表达于24～48h开始下降．72h趋于接种前水平,而TLR2 mRNA表达则持续增加至72h。结论：白念珠菌感染可迅速上调局部组织TLR2、TLR4的基因表达,TLR2、TLR4在口腔黏膜抗白念珠菌感染早期免疫中可能起重要作用。     

阴道念珠菌病患者体内白念珠菌抗氧化基因的表达

目的检测阴道念珠菌病患者体内感染状态下白念珠菌抗氧化基因的表达,初步了解抗氧化机制在阴道念珠菌病发病机制中的关系。方法选取7例阴道念珠菌病患者阴道分泌物分离株,培养鉴定为白念珠菌,选取PDA斜面培养3天(体外培养静止期)及临床阴道分泌物检体,用Trizol法提取总RNA反转录为cDNA后进行荧光定量PCR,检测体外培养静止期与体内检体抗氧化基因(CAT,SOD2,SOD5)及抗氧化转录因子(CaSkn7,CaHog1,CAP1)的基因表达情况,运用Ct值比较法进行表达量的相对定量分析。结果白念珠菌阴道病患者体内标本中CaSkn7,CAP1,SOD2,SOD5基因表达量比体外培养静止期少,分别为体外表达量的0.273,0.635,0.039,0.632;而CaHog1及CAT基因表达比体外培养静止期高,分别为体外表达量的3.5倍及1.67倍。经Wilcoxon检验,显示阴道分泌物标本中仅CaSkn7基因表达量较体外培养静止期低,差异有统计学意义P0.05,其余基因与体外培养静止期差异无统计学意义,P均0.05。结论白念珠菌黏膜感染状态下抗氧化相关的基因表达较少,抗氧化机制在念珠菌性阴道炎发病过程中作用不明显。     

白念珠菌基因组结构与遗传、致病相关基因

白念珠菌是一种重要的机会致病菌,在人的多个系统或器官与宿主共栖生存,可导致口腔和阴道念珠菌病,在免疫力低下或患严重疾病的患者,可引起系统疾病.由于早期准确诊断系统性念珠菌病存在困难,合适有效的抗真菌药物数量有限,病原体的耐药性增加,导致系统性念珠菌病病死率居高不下.因此,对白念珠菌基因组序列及结构、功能的研究对指导临床的诊断和治疗显得尤为重要.