健康人群口腔酵母菌的分离及菌种分布

目的调查健康人群口腔酵母菌的分布。方法将来源于健康人的口腔拭子接种于改良SDA培养基,37℃培养14d,分到的酵母菌通过菌落形态、革兰染色作初步鉴定．芽管形成试验、厚膜孢子生成及假菌丝产生试验、YBC酵母鉴定卡作菌种鉴定。结果酵母菌在健康人口腔中总分离率为8．78％(86／979),其中较多的为白色假丝酵母菌37株,占43．02％,近平滑假丝酵母菌和季也蒙假丝酵母菌各9株,各占10．47％,葡萄牙假丝酵母菌6株,占6．98％,其他25株。结论健康人口腔中有多种条件致病酵母菌的寄生。     

复发性外阴阴道假丝酵母菌病菌种分型及药敏

目的对RVVC、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RVVC发生的原因。方法用沙保弱琼脂培养基培养阴道分泌物,分离纯化菌株;VITEK 2全自动微生物分析仪鉴定菌种;同时进行药物敏感试验。结果共分离出101株假丝酵母菌,其中RVVC组45株,VVC组51株,健康组5株。101株中白色假丝酵母菌86株,占85.1%(86/101),RVVC组45株中35株为白色假丝酵母菌,占77.8%(35/45);VVC组51株中47株为白色假丝酵母菌,占92.2%(47/51),健康组5株中4株为白色假丝酵母菌,占80%(4/5)。RV-VC组非白色假丝酵母菌比例高,与VVC组比较差异有显著性(P<0.05)。RVVC组假丝酵母菌对唑类药物敏感性低于VVC组。结论VVC、RVVC的主要致病菌仍是白色假丝酵母菌,RVVC组非白色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率明显低于VVC组。     

医院内深部真菌感染分布及耐药性分析

目的探讨大连医科大学附属第一医院深部真菌感染的常见菌种,临床病区分布及其耐药性。方法对2008年1月1日至2008年12月31日749例住院患者深部真菌感染进行分析。结果常见菌种为白色假丝酵母菌,热带假丝酵母菌,近平滑假丝酵母菌,克柔假丝酵母菌,光滑球拟酵母菌。主要病区为ICU。从标本的来源看,痰标本中真菌检出率最高,占61.1%。两性霉素B抗菌作用最强,耐药率只有3.6%。结论医院内深部真菌感染分布广泛,非白色假丝酵母菌感染明显增多,耐药菌株增多,部分治疗需联合用药。     

慢性阻塞性肺疾病患者继发真菌性医院肺炎的病原菌分布及耐药性分析

目的探讨慢性阻塞性肺疾病（Chronic obstructive pulmonary diseases,COPD）患者继发真菌性医院肺炎的病原菌分布及耐药情况。方法回顾性分析2011年1月至2012年12月间31例COPD患者继发肺部真菌感染的情况。结果 31例COPD继发肺部真菌感染的患者送检标本中共检到34株真菌,以曲霉菌属居首位,占29.41%,其次为白色假丝酵母菌,占26.41%。7种常用抗真菌药物对真菌显示有不同的抗菌活性,尤以伊曲康唑、氟康唑、5-氟胞嘧啶为佳。结论 COPD患者继发肺部真菌感染多为曲霉菌属和白色假丝酵母菌。临床疑为COPD患者继发肺部真菌感染病例须及时采集标本培养,以早期发现病原性真菌和选择有效药物治疗。     

海南岛4市县人群口腔假丝酵母菌分布情况的初步调查

目的：调查海南岛海口、三亚市、五指山市和保亭县四市县人群口腔假丝酵母菌的分布情况,为口腔假丝酵母菌分子生物学和遗传学特点研究提供基础资料。方法：分离出的假丝酵母菌经过菌落特征、制片乳酸酚棉蓝染色、芽管形成试验、厚膜孢子形成试验等初步鉴定,并用科玛嘉显色培养基培养确定。结果：海口、三亚市、五指山市和保亭县人群口腔假丝酵母菌总分离率为25．23％．其中白假丝酵母菌占54．48％,光滑假丝酵母菌占14．04％,热带假丝酵母菌占11．06％,克柔假丝酵母菌占9．36％。住院病人和健康人口腔假丝酵母菌的分离率分别为39．95％和11．61％,差异有统计学意义（P〈0．01）,结论：海南地区人群口腔假丝酵母菌的总分离率高于国内其它省份,分离出的假丝酵母菌以白假丝酵母菌为首位,病人口腔假丝酵母菌分离率明显高于健康人。     

男性生殖道真菌感染菌种构成及耐药分析

目的探讨男性生殖道真菌感染病原菌的种类构成及耐药性,为临床治疗提供依据。方法采集2367份男性患者生殖道分泌物立即送检采用法国生物梅里埃公司Vitek2-Compact进行菌种鉴定以及ATB FUNGUS3真菌药敏试剂条进行药敏试验。结果 2 367例男性生殖道炎症患者中,共检出真菌410例,检出率为17.32%,其中白色假丝酵母菌303株,占73.90%;近平滑假丝酵母菌75株,占18.29%;热带假丝酵母菌16株,占3.90%;4种假丝酵母菌对两性霉素B的敏感率为100.00%;白色假丝酵母菌和热带假丝酵母菌对伏立康唑、氟康唑、依曲康唑耐药率分别为1.65%、1.32%、7.59%和12.50%、12.50%、18.75%。结论假丝酵母菌在男性生殖道感染中占有一定比例;菌种以白色假丝酵母菌为主,近平滑假丝酵母菌次之。因为它们对唑类抗真菌药物有不同程度的耐药,所以临床应加强假丝酵母菌的检测和药敏分析,合理选用药物。     

慢性前列腺炎患者假丝酵母菌感染及耐药性分析

目的 了解慢性前列腺炎与假丝酵母菌感染的相关性及假丝酵母菌的耐药性。方法 采用常规沙堡平板分离360例慢性前列腺炎患者的前列腺液标本中的假丝酵母菌,疑似菌落用ATB Expression鉴定仪进行鉴定。采用ATB-Fungus真菌药敏板,对假丝酵母菌株进行药敏试验。结果 11．7％前列腺液标本（42／360）似丝酵母菌阳性,其中自假丝酵母菌23例（54．7％）,近平滑假丝酵母菌13例（30．9％）,其它6例（14．3％）。假丝酵母菌菌株对两性霉素B和制霉菌素敏感率均为100％,其次为酮康唑,敏感率为97．0％;对5-氟胞嘧啶耐药性最强,其耐药率为56．5％。结论 白假丝酵母菌和近平滑假丝酵母菌是慢性前列腺炎假丝酵母菌感染的优势菌种,假丝酵母菌株最敏感的药物是两性霉素B和制霉菌素。     

2型糖尿病患者真菌性泌尿系感染的真菌菌群分布及耐药性分析

目的探讨2型糖尿病患者真菌性泌尿系感染的真菌菌群分布及耐药性。方法回顾性分析在台州市恩泽医疗中心(集团)路桥医院就诊的1 000例2型糖尿病患者,所有患者均送检清洁中段尿进行检查,对判断为真菌性泌尿系感染患者的尿液进行真菌培养,观察菌群分布及耐药性。结果 2型糖尿病患者泌尿系真菌感染率达8.70%,共培养出103株真菌,其中排名前六的真菌分别是白假丝酵母菌、光滑假丝酵母菌、热带假丝酵母菌、克柔假丝酵母菌、法氏假丝酵母菌和近平滑假丝酵母菌;对主要致病菌耐药性较低的药物是5-氟胞嘧啶(2.91%)和两性霉素B(0.97%)。两性霉素B和5-氟胞嘧啶的耐药率均明显低于氟康唑、伊曲康唑与伏立康唑(χ~2=21.58、31.93;17.94、16.43;26.30、13.03,P0.01)。结论 2型糖尿病患者真菌性泌尿系感染的发生率较高,以白假丝酵母菌、光滑假丝酵母菌和热带假丝酵母菌最为多见,临床医师应根据病原学、耐药性检查及安全性选择合适的抗生素,对无法进行耐药性检查的患者可首选两性霉素B。     

血流感染真菌的分布及耐药性分析

目的了解真菌性血液感染常见病原真茵的菌群分布及耐药性状况,为临床真菌感染性疾病提供病原学诊断和合理使用抗真菌药物的依据。方法回顾性分析浙江大学医学院附属第一医院5年（2008-2012年）间导致真菌性血液感染的217株真菌的菌群分布及药物敏感性状况。结果217株真菌标本中以假丝酵母菌属为主,占88. 5%;其中又以白假丝酵母菌比例最高,占35.4%。菌种分布与患者的性别和年龄存在相关性。60-79岁年龄组感染白假丝酵母菌多见,占49% ,这一比例明显高于其他真菌。男性更容易感染白假丝酵母菌（40.1% vs 24.6%,P〈0.05）,女性感染近平滑假丝酵母菌更常见（24.6% vs 11.8%,P〈0.05）.假丝酵母菌和新生隐球菌对两性霉素B、伏立康唑、氟康唑、伊曲康唑、5-氟胞嘧啶的敏感性均较高。光滑假丝酵母菌对伊曲康唑敏感率较低,仅为60.0%。结论真菌血流感染在临床呈上升趋势,抗真菌药物均体现较高抗菌活性,对真菌感染进行地区层面流行病学调查和耐药监测很重要。     

泌尿系感染真菌菌种分布及耐药性分析

目的 了解引起泌尿系感染真菌菌种分布及其耐药特点。方法 用沙保弱培养基进行真菌分离培养,鉴定用API20CAUX生化系列,药敏用ATBFUNGUS药敏板。结果427株真菌中前5位为白色假丝酵母菌215株、热带假丝酵母菌69株、伪热带假丝酵母菌38株、克柔假丝酵母菌27株、光滑球拟酵母菌23株。5-氟胞嘧啶、两性霉索B、制霉菌素、咪康唑、益康唑、酮康唑的敏感性分别为90．2％、90．2％、89．2％、73．8％、71．4％和62．9％。临床感染真菌种类及其耐药性有逐年上升趋势。结论 引起泌尿系感染的真菌广泛存在,但仍以白色假丝酵母菌为主。对5-氟胞嘧啶、两性霉索B、制霉菌素敏感率较高;唑类抗真菌药的敏感性呈明显下降趋势。     

N-cadherin mediates endocytosis of Candida albicans by endothelial cells

Candida albicans is the most common cause of fungal bloodstream infections. To invade the deep tissues, blood-borne organisms must cross the endothelial cell lining of the vasculature. We have found previously that C. albicans hyphae, but not blastospores, invade endothelial cells in vitro by inducing their own endocytosis. Therefore, we set out to identify the endothelial cell receptor that mediates the endocytosis of C. albicans. We determined that endocytosis of C. albicans was not mediated by bridging molecules in the serum and that it was partially dependent on the presence of extracellular calcium. Using an affinity purification procedure, we discovered that endothelial cell N-cadherin bound to C. albicans hyphae but not blastospores. N-cadherin also co-localized with C. albicans hyphae that were being endocytosed by endothelial cells. Chinese hamster ovary (CHO) cells expressing human N-cadherin endocytosed significantly more C. albicans hyphae than did CHO cells expressing either human VE-cadherin or no human cadherins. The expression of N-cadherin by the CHO cells resulted in enhanced endocytosis of hyphae, but not blastospores, indicating the selectivity of the N-cadherin-mediated endocytosis. Down-regulation of endothelial cell N-cadherin expression with small interfering RNA significantly inhibited the endocytosis of C. albicans hyphae. Therefore, a novel function of N-cadherin is that it serves as an endothelial cell receptor, which mediates the endocytosis of C. albicans.     

Septin function in Candida albicans morphogenesis

The septin proteins function in the formation of septa, mating projections, and spores in Saccharomyces cerevisiae, as well as in cell division and other processes in animal cells. Candida albicans septins were examined in this study for their roles in morphogenesis of this multimorphic, opportunistically pathogenic fungus, which can range from round budding yeast to elongated hyphae. C. albicans green fluorescent protein labeled septin proteins localized to a tight ring at the bud and pseudohyphae necks and as a more diffuse array in emerging germ tubes of hyphae. Deletion analysis demonstrated that the C. albicans homologs of the S. cerevisiae CDC3 and CDC12 septins are essential for viability. In contrast, the C. albicans cdc10Delta and cdc11Delta mutants were viable but displayed conditional defects in cytokinesis, localization of cell wall chitin, and bud morphology. The mutant phenotypes were not identical, however, indicating that these septins carry out distinct functions. The viable septin mutants could be stimulated to undergo hyphal morphogenesis but formed hyphae with abnormal curvature, and they differed from wild type in the selection of sites for subsequent rounds of hyphal formation. The cdc11Delta mutants were also defective for invasive growth when embedded in agar. These results further extend the known roles of the septins by demonstrating that they are essential for the proper morphogenesis of C. albicans during both budding and filamentous growth.     

Candida albicans infection inhibits macrophage cell division and proliferation

The pathogenicity of the opportunistic human fungal pathogen Candida albicans depends on its ability to inhibit effective destruction by host phagocytes. Using live cell video microscopy, we show here for the first time that C. albicans inhibits cell division in macrophages undergoing mitosis. Inhibition of macrophage cell division is dependent on the ability of C. albicans to form hyphae, as it is rarely observed following phagocytosis of UV-killed or morphogenesis-defective mutant Candida. Interestingly, failed cell division following phagocytosis of hyphal C. albicans is surprisingly common, and leads to the formation of large multinuclear macrophages. This raises question as to whether inhibition of macrophage cell division is another virulence attribute of C. albicans or enables host macrophages to contain the pathogen.     

Identification of salivary components that induce transition of hyphae to yeast in Candida albicans

Candida albicans , the major human fungal pathogen, undergoes a reversible morphological transition from single yeast cells to pseudohyphae and hyphae filaments. The hyphae form is considered the most invasive form of the fungus. The purpose of this study is to investigate the effect of saliva on hyphae growth of C. albicans. Candida albicans hyphae were inoculated in Roswell Park Memorial Institute medium with whole saliva, parotid saliva or buffer mimicking the saliva ion composition, and cultured for 18 h at 37 °C under aerobic conditions with 5% CO₂. Whole saliva and parotid saliva induced transition to yeast growth, whereas the culture with buffer remained in the hyphae form. Parotid saliva was fractionated on a reverse-phase C8 column and each fraction was tested for inducing transition to yeast growth. By immunoblotting, the salivary component in the active fraction was identified as statherin, a phosphoprotein of 43 amino acids that has been implicated in remineralization of the teeth. Synthetically made statherin induced transition of hyphae to yeast. By deletion of five amino acids at the negatively charged N-terminal site (DpSpSEE), yeast-inducing activity and binding to C. albicans were increased. In conclusion, statherin induces transition to yeast of C. albicans hyphae and may thus contribute to the oral defense against candidiasis.     

布拉氏酵母菌治疗小儿轮状病毒肠炎的Meta分析

目的系统评价布拉氏酵母菌治疗小儿轮状病毒肠炎的疗效和安全性。方法在万方数据知识平台、中国知网、中国生物医学文献数据库、维普中文科技期刊数据库、PubMed、Web of Science、Embase、Cochrane图书馆等中检索布拉氏酵母菌治疗小儿轮状病毒肠炎的相关文献,并追索已纳入的参考文献。对文献进行评价与资料提取,使用Stata 14.0软件进行Meta分析。结果纳入12篇文献,共1 233例患儿(626例为试验组,607例为对照组),7篇文献采用止泻时间进行评价,显示布拉氏酵母菌能够缩短小儿轮状病毒肠炎腹泻时间[SMD=-0.73,95%CI(-1.05,-0.41),I~2=75.7%];5篇文献以住院时间进行评价,显示布拉氏酵母菌能够缩短小儿轮状病毒肠炎的住院时间[SMD=-0.99,95%CI(-1.24,-0.74),I~2=56.5%];10篇文献以总有效率进行评价,显示布拉氏酵母菌能够提高小儿轮状病毒肠炎的总有效率[RR=1.26,95%CI(1.19, 1.34),I~2=26.2%];仅有1篇文献报道了轻微的不良反应。结论布拉氏酵母菌治疗小儿轮状病毒肠炎疗效显著,且不良反应轻微,值得临床推广应用。     

Structural characterization of (1-->3)-beta-D-glucans isolated from blastospore and hyphal forms of Candida albicans

Glucans are (1-->3)-beta-linked linear and branched polymers containing anhydroglucose repeat units. They comprise a major portion of the cell wall of saprophytic and pathogenic fungi. Glucans activate a wide range of innate immune responses. They are also released from the fungal cell wall as exopolymers into the blood of patients with fungal infections. Extensive studies have been done on glucans isolated from saprophytic fungi, such as Saccharomyces cerevisiae; however, much less is known about the glucans produced by the polymorphic fungal pathogen Candida albicans. We have undertaken an extensive structural characterization and comparison of glucans isolated from C. albicans blastospores and hyphae using high-resolution, solution-state proton nuclear magnetic resonance spectroscopy (NMR). In addition, we developed a simple and straightforward method for the production of Candida hyphae that resulted in gram quantities of hyphal mass. Also, we compared and contrasted the Candida glucans isolated by two different protocols with those isolated from S. cerevisiae. Isolation protocols provide high purity glucans with source-based structural differences. Structural details provided by this NMR analysis included the degree of polymerization, molecular weight, degree and type of branching, and structural composition. We observed that Candida glucans, derived from blastospores or hyphae, are different compared to those isolated from S. cerevisiae with regard to side-chain branching along the backbone and at the reducing terminus. These structural details are an important prerequisite for biomedical studies on the interaction of isolated fungal cell wall glucans with the innate immune system.     

Arbuscular mycorrhizal structure and fungi associated with mosses

We investigated the colonization and diversity of arbuscular mycorrhizal (AM) fungi associated with 24 moss species belonging to 16 families in China. AM fungal structures, i.e. spores, vesicles, hyphal coils (including intracellular hyphae), or intercellular nonseptate hyphae, were found in 21 moss species. AM fungal structures (vesicles, hyphal coils, and intercellular nonseptate hyphae) were present in tissues of 14 moss species, and spores and nonseptate hyphae on the surface of gametophytes occurred in 15 species. AM fungal structures were present in 11 of the 12 saxicolous moss species and in six of the ten terricolous moss species, but absent in two epixylous moss species. AM fungal structures were only observed in moss stem and leaf tissues, but not in rhizoids. A total of 15 AM fungal taxa were isolated based on trap culture with clover, using 13 moss species as inocula. Of these AM fungi, 11 belonged to Glomus, two to Acaulospora, one to Gigaspora, and one to Paraglomus. Our results suggest that AM fungal structures commonly occur in most mosses and that diverse AM fungi, particularly Glomus species, are associated with mosses.     

LAAE-14, a new anti-inflammatory drug, increases the survival of Candida albicans-inoculated mice

LAAE-14, a lipidic acid-amido ether derivative, has been recently described as a new anti-inflammatory drug. We have studied the effect of treatment with this compound on the susceptibility of mice to in vivo experimental Candida albicans infection. ICR mice orally treated with LAAE-14 (25 mg kg(-1)) and experimentally intravenously infected showed a significantly increased survival as compared to control mice. In vitro, the compound did not inhibit the growth of C. albicans yeast cells or the yeast-to-hyphal transition. The in vitro production of prostaglandin E2 by peritoneal macrophages in response to the yeasts and hyphae of C. albicans was significantly decreased upon treatment with LAAE-14, in a dose-dependent manner. Thus, reduced prostaglandin production during fungal infection could be an important factor in controlling fungal colonisation and infection.     

Effects ofN,N-bis(3-aminopropyl)dodecylamine on antioxidant enzyme activities, mitochondrial morphology and metabolism inAspergillus niger

The activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSP) as well as of succinate dehydrogenase (SDG), NADH dehydrogenase (NDG) and fumarate hydratase (FHT) were examined in relation to mitochondrial ultrastructure changes in Aspergillus niger exposed to N,N-bis(3-aminopropyl)dodecylamine (Apd) that was shown to exhibit fungicidal activity. There was a progressive increase in SOD, CAT and GSP activities 1 and 4 h after 0.05 and 0.1 % Apd application. However, this was followed by a pronounced activity decrease when 0.05 % Apd treatment was prolonged by 1 d. The destructive effect on fungal morphology was observed when this fungicidal agent was applied at the concentration of 0.1 % for 1 d. In the treated hyphae mitochondria degenerated after all organelles. The morphological malformations of mitochondria had an impact on their metabolic state; however, the activities of SDG, NDG and FHT were affected to a different extent. In A. niger the fungicidal effect of Apd could be mediated by oxidative stress impairing the vital mitochondria-related cellular functions.