念珠菌属的分布及其对五种抗真菌药物的耐药性分析

目的检测念珠菌对5种抗真菌药物体外敏感性，了解念珠菌耐药性变化趋势。方法用沙氏培养基进行真菌分离，采用念珠菌显色培养基作菌种鉴定，用ATBFUNGUS3药敏试剂盒测定抗真菌药物对临床分离的念珠菌最低抑菌浓度（MIC）。结果白色念珠菌的耐药率为最低，它除对伊曲康唑耐药率为10．8％外，其余均低于5．4％；而以克柔念珠菌的耐药率最高，它对氟康唑和伊曲康唑的耐药率大于33.3％。结论不同念珠菌对常用抗真菌药物耐药性存在差异，白色念珠菌对抗真菌药物的耐药性要低于非白色念珠菌。因此真菌的培养鉴定和药敏试验对临床合理选择抗真菌药物十分重要。     

白色念珠菌对常用抗真菌药物的耐药性分析

近年来,随着大量广谱抗菌药、免疫抑制剂和皮质激素及介入性检查的广泛应用,导致临床标本中酵母样真菌的分离率显著增加,由酵母样真菌引起的感染率逐渐升高,加之抗真菌药物的开发和在临床上的广泛使用,从而又导致耐药株的产生。目前,酵母样真菌（白色念珠菌）对临床上常用的几种抗真菌药物的耐药率已达到相当高的比例,这给临床上抗真菌感染治疗带来一定困难。为了解白色念珠菌对抗真菌药物的敏感性。以便为临床正确选择抗真菌药物和有效控制白色念珠菌引起的感染提供依据,黄石市四医院测定了五种常用抗真菌药物对89株白色念珠菌最低抑菌浓度（MIC）,现报告如下。     

耐氟康唑阴道白色念珠菌ERG基因家族突变研究及唑类药物抑菌分析

目的研究耐氟康唑阴道白色念珠菌ERG11、ERG3和ERG5基因突变,及分离株对酮康唑、克霉唑、硝酸咪康唑的抑菌分析。 方法收集221株经质谱仪鉴定为白色念珠菌的菌株,微量肉汤稀释法进行氟康唑、酮康唑、克霉唑、硝酸咪康唑药敏试验,得出四种药物对白色念珠菌最小抑菌浓度（MIC）值;根据CLSI M27-A3筛选出氟康唑非敏感白色念珠菌,提取DNA后扩增ERG11、ERG3与ERG5基因,进行双向测序,与GenBank中的标准菌株基因序列（SC5314和AF069752）进行BLAST比对分析。 结果白色念珠菌对氟康唑耐药率为9.05%;对白色念珠菌MIC值进行比较,酮康唑的MIC值均低于克霉唑和硝酸咪康唑,差异有统计学意义（P＜0.05）。克霉唑与硝酸咪康唑比较,差异无统计学意义（P＞0.05）。ERG11、ERG3、ERG5分别发现18、17、12种错义突变。 结论酮康唑对白色念珠菌抑菌率总体比克霉唑、硝酸咪康唑强。耐氟康唑白色念珠菌ERG11、ERG3、ERG5基因出现若干错义突变位点,某些位点突变导致氨基酸变异可能与耐药产生有关。     

探讨五倍子提取物没食子酸抑制白色念珠菌作用机制

目的 探讨五倍子提取物没食子酸对抑制白色念珠菌所发挥的作用。方法 通过水提取法及正交设计安排实验方法分析五倍子提取物没食子酸提取含量相关影响因素并分析对白色念珠菌的抑菌效果。结果 随着提取次数增加以及单次提取时间延长,没食子酸含量随之增加,pH值下降,抑菌群直径增大,试验号1号、2号、3号、4号实验效果较试验号5号及6号明显。没食子酸在490nm处吸光度（OD490nm）与浓度线性关系良好,测定五倍子提取物没食子酸吸光度值并依照没食子酸标准曲线回归方程为y=1.3799χ-0.0163可得五倍子提取物没食子酸浓度为4.17g/L时抗白色念珠菌效果最为理想。结论 固定加水倍数并增加提取次数,延长提取时间,可有效提高提取液中没食子酸含量且pH值越低,没食子酸针对白色念珠菌的抑菌作用可得到增强。     

白色念珠菌CDR基因表达与氟康唑体外药敏试验的关系研究

目的调查白色念珠菌临床株对氟康唑的耐药情况,研究白色念珠菌临床耐药基因的表达与氟康唑耐药之间的关系。方法采用M27-A2微量肉汤稀释法,对某院微生物实验室2010年分离保存的221株来自就诊患者的痰、中断尿、粪便和咽拭子的白色念珠菌进行氟康唑MIC值测定。以白色念珠菌18S RNA为内参照,采用RT-PCR技术观察比较耐药株和敏感株组在不同氟康唑浓度下CDR1和CDR2基因的转录水平。结果 221株白色念珠菌中,有57.47%（127株）对氟康唑敏感,10.86%（24株）剂量依赖敏感,31.67%（70株）耐药。不同药物浓度作用下,耐药株组CDR1和CDR2的表达量相对要高于敏感株组（P〈0.05）。结论白色念珠球菌对氟康唑耐药率较高,耐药株组CDR1和CDR2表达量高于敏感株组。     

不同培养时间与培养基pH值对体外实验中喹诺酮类抗解脲脲支原体药物敏感性的影响

目的研究不同培养时间及培养基pH值对左氧氟沙星、司帕沙星及加替沙星在体外抗解脲脲支原体（Uu）MIC的影响。方法用微量液体倍比稀释法,测定了35株临床Uu对3种药物在pH值6.0、6.5、7.0的培养基中,分别培养24、36、48、60 h时的MIC并统计分析。结果60 h的MIC与48 h相同,pH值〉6.5时,Uu对加替沙星100.0%敏感;司帕沙星耐药率较高;在不同pH值时,培养时间由24 h延长到36 h及48 h,左氧氟沙星、司帕沙星及加替沙星的MIC分别可增加1-32、1-8及1-16倍（各组P〈0.05）,耐药率也逐渐升高;pH值由6.0升高到6.5及7.0时,在不同培养时间,司帕沙星及加替沙星的MIC均可降低1-8倍（各组P〈0.05）,耐药率均逐渐降低;而左氧氟沙星的MIC50、MIC90及耐药率均升高。结论体外实验中,司帕沙星及加替沙星的MIC随培养时间延长而升高,随培养基pH值升高而降低;左氧氟沙星的MIC随培养时间延长及pH值升高而升高;加替沙星有良好的抗Uu活性;培养48 h可以作为测定MIC的终点时间。     

科玛嘉念珠菌显色培养基的临床应用及评价

科玛嘉念珠菌显色培养基是一种新型分离培养基,根据培养基对不同酵母样真菌菌落颜色及形态差异,可以选择性地分离念珠菌并同时对白色念珠菌、热带念珠菌、光滑念珠菌和克柔念珠菌作出鉴定.我们对605例真菌培养标本分离出的329株酵母样真菌同时使用科玛嘉念珠菌显色培养基和梅里埃YBC鉴定卡进行鉴定,以评价科玛嘉念珠菌显色培养基在临床酵母样真菌分离与鉴定方面的应用价值.     

用于白色念珠菌快速鉴定的两种培养基

白色念珠菌为临床上最为常见、致病力也最强的酵母样真菌,近年来由其引起的感染逐渐增加,成为当前较棘手的医学问题之一,因而从食品、环境及各种临床标本中进行白色念珠菌的快速分离和鉴定显得甚为重要。我们采用TTC-玉米吐温琼脂培养基（TCTA）和苯胺蓝酵母培养基（ABYP）用于该菌的快速鉴定,取得较满意效果,报告如下。1 材料与方法1.1 菌株 共156株酵母样真菌,系从临床标本或环境中分离而来,其中白色念珠菌85株,其它念珠菌66株,新生隐球菌5株,均经系统鉴定无误。1.2 TCTA培养基 玉米吐温培养基（玉米粉5％,琼脂1.5％,吐温-801％）经 121℃灭菌20min后加入终含量为0.02％的     

几种沙氏培养基真菌生长情况比较

目的了解沙氏培养基对真菌检测结果的影响,为一次性使用卫生用品实验室检测提供依据。方法选7种不同厂家生产的沙氏培养基（含氯霉素与否）用0.9%生理盐水倍比稀释后进行真菌菌落总数检测。结果白色念珠菌（CMCC10231）、黑曲霉菌（ATCC16404）和酵母菌（ATCC18790）在7种培养基各稀释度平皿上菌落生长速度、菌落形态及大小、菌落总数基本一致;金黄色葡萄球菌（ATCC260039）和大肠杆菌（ATCC25922）在含氯霉素沙氏培养基上生长均受抑制。结论一次性使用卫生用品真菌菌落总数检测应选用含氯霉素的沙氏琼脂培养基,抑制细菌生长,提高实验室的准确度。     

435株真菌的临床分布及药敏分析

目的了解临床分离的真菌分布,分析其药敏性,为临床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。方法收集2010年1-12月中南大学湘雅三医院住院患者送检标本,经血培养基或沙保罗培养基培养后,挑取酵母样菌落,进行菌种鉴定和药敏试验。结果分离到435株真菌,居前三位者分别为白色念珠菌（54.9%）,近平滑念珠菌（20.5%）,葡萄牙念珠菌（11.3%）。这些真菌主要来自重症监护室（ICU）（32.6%）和痰标本中（74.8%）。药敏结果显示,分离的酵母样真菌对5-氟胞嘧啶、两性霉素B和酮康唑敏感性较高,分别为97.5%、96.8%和95.8%。对氟康唑、伊曲康唑和益康唑耐药性较高,分别为47.4%、43.0%和21.2%。结论真菌临床分离菌株以白色念珠菌和近平滑念珠菌为主,临床真菌对5-氟胞嘧啶、两性霉素B和酮康唑具有较高的敏感性。     

80株生殖道白念珠菌的药物敏感性研究

目的 为了解生殖道白念珠菌对五种抗真菌药物的敏感性及是否存在交叉耐药性，了解对抗真菌药物的耐药率，使临床医师能选择敏感的药物控制白念珠菌感染。方法 采用NCCLS M27-A推荐方法检测80株临床分离的生殖道白念珠菌对五种抗真菌药物的最低抑菌浓度(MIC80)。结果 80株临床分离生殖道白念珠菌对五种抗真菌药物的敏感率分别从41．3％至87．5％不等，而耐药率从0％至26．2％不等。且10株同时对氟康唑、咪康唑耐药，2株同时对氟康唑、酮康唑耐药，有1株同时对咪康唑、酮康唑耐药。结论 发现白念珠菌对两性霉素B最敏感，而对唑类药物耐药率较高，且唑类药物之间存在交叉耐药性。     

Synthesis of novel triazole derivatives as inhibitors of cytochrome P450 14alpha-demethylase (CYP51)

A series of 1-(1H-1,2,4-triazol-1-yl)-2-(2,4-difluorophenyl)-3-[(4-substitutedphenyl)-piperazin-1-yl]-propan-2-ols have been designed and synthesized on the basis of the structure-activity relationships and antimycotic mechanism of azole antifungal agents. Their structures were confirmed by elemental analysis, IR, MS and (1)H NMR. Results of preliminary antifungal tests against eight human pathogenic fungi (Candida albicans, Candida parapsilosis, Candida tropicalis, Cryptococcus neoformans, Aspergillus fumigatus, Trichophyton rubrum, Fonsecaea compacta, and Microsporum gypseum) in vitro showed that all title compounds exhibited activity against fungi tested to some extent. Among the compounds tested, all compounds showed higher activity against C. albicans than fluconazole in vitro. Compounds 3, 6-8, 28, 29, and 32 exhibited the same activities against C. albicans as voriconazole (with the MIC value of 0.0152microg/mL). Compounds 3, 6, and 7 showed higher activity against C. parapsilosis than all five positive controls.     

Anti-Candida activity of Mentha arvensis and Turnera ulmifolia

Candidiasis is the most frequent infection by opportunistic fungi, frequently caused by Candida albicans, Candida tropicalis, Candida parapsilosis, Candida glabrata, and Candida krusei. Mentha arvensis L. is a herbaceous plant that occurs throughout South America and is used as a tea and in the folk medicine. Turnera ulmifolia L. is already known to be of medicinal value. Ethanol extracts from M. arvensis and T. ulmifolia were assayed for antifungal activity against strains of C. albicans, C. tropicalis, and C. krusei. No clinically relevant antifungal activity was demonstrated by the extracts; however, a potentiation effect was observed when the extracts were applied with metronidazole against C. tropicalis. M. arvensis and T. ulmifolia could represent a source of natural products with modifying antifungal activity.     

铜绿假单胞菌代谢产物体外对白假丝酵母菌等的抑菌活性研究

目的观察铜绿假单胞菌代谢产物对假丝酵母菌属的体外抑菌活性。方法用交叉条带实验方法测定铜绿假单胞菌对54株假丝酵母菌属的体外抑制活性。结果铜绿假单胞菌产生的抗菌物质对白假丝酵母菌及光滑假丝酵母菌的抑菌作用最强,抑菌带最宽。产蓝绿色素的第6株铜绿假单胞菌对白假丝酵母菌和热带假丝酵母菌的抑制率均达100%,产黄绿色素的第8株铜绿假单胞菌对白假丝酵母菌、热带假丝酵母菌和克柔假丝酵母菌的抑制率也均达100%,铜绿假单胞菌产色素菌株的抗真菌活性优于不产色素的菌株。结论铜绿假单胞菌产生的抗菌物质对白假丝酵母菌、光滑假丝酵母菌具有较强的抗真菌活性。     

Testing chemical germicides against Candida species using quantitative carrier and fingerpad methods

Six disinfectants were tested against Candida albicans, C. parapsilosis and C. tropicalis using quantitative carrier tests based on glass (QCT-1) and metal (QCT-2) surfaces. C. albicans was also used to test four topical agents by a fingerpad method. Hard water (200 ppm as CaCO(3)) was the product diluent. In preliminary tests with QCT-1 and QCT-2, the testing was with or without a soil load; subsequent tests and fingerpad tests included soil. In QCT-1 and QCT-2, each carrier received 10 microL (5.0 x 10(6) - 1.0 x 10(7)colony forming units) of Candida, and was air dried for 1 h, then exposed to 1 mL or 50 microL of test product at 22 +/- 2 degrees C for up to 10 min. Controls received an equivalent volume of saline. For fingerpad tests, each digit received 10 microL of inoculum, which was allowed to dry and exposed to 1 mL of test product for 20 s. Inoculated plates of Sabouraud's dextrose agar were held for 48 h at 30 degrees C and colonies counted to determine reductions in colony forming units. In tests on both hard surfaces and fingerpads, ethanol and products based on ethanol reliably and rapidly inactivated all the Candida species tested. Products with sufficient potency to have tuberculocidal claims produced substantial reductions in the titre of C. albicans, although some showed a lesser reduction in titre of C. tropicalis and C. parapsilosis. This may reflect differences in cell hydrophobicity between Candida species, and highlights the need for care in selecting a suitable surrogate for disinfectant tests. The quantitative carrier and fingerpad protocols are suitable for assessing the activity of disinfectants and topical antiseptics against candida.     

Synthesis, antimicrobial activity, pharmacophore analysis of some new 2-(substitutedphenyl/benzyl)-5-[(2-benzofuryl)carboxamido]benzoxazoles

The synthesis and antimicrobial activity of a new series of 2-(substitutedphenyl/benzyl)-5-[(2-benzofuryl)carboxamido]benzoxazole derivatives 3-12 were described. The in vitro antimicrobial activity of the compounds was determined against some Gram-positive, Gram-negative bacteria and fungi and their drug-resistant isolates in comparison with standard drugs. Antimicrobial results indicated that the synthesized compounds possessed a broad spectrum of activity with MIC values 500-15.625 microg/ml. In the series, the most active compound against Candida krusei and Candida albicans isolate is 8 with MIC value 31.25 microg/ml. However, it is one dilution less potent than the compared fluconazole. Some of the screened compounds exhibit significant activity, having MIC value as 31.25 microg/ml in Pseudomonas aeruginosa having same activity as Rifampicin. Furthermore, considering the worth of developing new antibacterial agents against drug-resistant P. aeruginosa the present study explores the structure-activity relationship analysis of 2-(substitutedphenyl/benzyl)-5-[(2-benzofuryl)carboxamido]benzoxazoles using 3D-common features pharmacophore hypotheses approach.     

A survey of the in vitro antifungal activity of heather (Erica sp.) organic honey

Monofloral heather (Erica sp.) honey samples (n=89), harvested in Portugal according to European organic beekeeping rules, were analyzed to test their antifungal effect against Candida albicans, Candida krusei, and Cryptococcus neoformans. A synthetic honey solution was also tested to determine antifungal activity attributable to sugars. The specific growth rate (μ) values showed that growth of all the yeasts was reduced in the presence of honey. The honey concentration (% wt/vol) that inhibited 10% of the yeast growth (X(min)) was 13.5% for C. albicans, 20.5% for C. krusei, and 17.1% for C. neoformans. The respective concentrations of heather honey and synthetic honey in the C. krusei culture medium above 60% (wt/vol) that inhibited 90% of the yeast growth (X(max)) and X(min), respectively, were established, whereas C. albicans and C. neoformans were more resistant because X(max) values were not reached over the range tested (10-60%, wt/vol). Heather honey might be tapped as a natural resource to look for new medicines for the treatment of mycotic infections. Further studies are now required to demonstrate if this antifungal activity has any clinical application.     

Synthesis and evaluation of novel 1-(1H-1,2,4-triazol-1-yl)-2-(2,4-difluorophenyl)-3-[(4-substitutedphenyl)-piperazin-1-yl]-propan-2-ols as antifungal agents

A series of 1-(1H-1,2,4-triazol-1-yl)-2-(2,4-difluorophenyl)-3-[(4-substitutedphenyl)-piperazin-1-yl]-propan-2-ols have been designed and synthesized on the basis of the structure-activity relationships and antimycotic mechanism of azole antifungal agents. Their structures were confirmed by elemental analysis, IR, MS, (1)H NMR and (13)C NMR. Results of preliminary antifungal tests against six human pathogenic fungi (Candida albicans, Candida parapsilosis, Cryptococcus neoformans, Candida tropicalis, inherently fluconazole-resistant Candida krusei, Candida glabrata) in vitro showed that all title compounds exhibited activity against fungi tested to some extent except against C. tropicalis. Compound 5b showed higher activity against C. albicans, C. parapsilosis and C. krusei than fluconazole, and its MIC values were determined to be 0.5microg/mL, 1microg/mL and 4microg/mL, respectively. Compound 5k showed higher activities against Torulopsis glabrata than fluconazole (with the MIC value of 2microg/mL). Compounds 5a, 5c, 5f, 5g, 5i exhibited higher activities against C. parapsilosis than fluconazole (with the MIC values of 2microg/mL, 2microg/mL, 2microg/mL, 1microg/mL and 2microg/mL, respectively).     

Synthesis and structure-activity relationships of new antimicrobial active multisubstituted benzazole derivatives

A series of multisubstituted benzoxazoles, benzimidazoles, and benzothiazoles (5-7) as non-nucleoside fused isosteric heterocyclic compounds was synthesized and tested for their antibacterial activities against various Gram-positive and Gram-negative bacteria and antifungal activity against the fungus Candida albicans. Microbiological results indicated that the synthesized compounds possessed a broad spectrum of activity against the tested microorganisms at MIC values between 100 and 3.12 microg/ml. Structure-activity relationships (SAR) studies revealed that benzothiazole ring system enhanced the antimicrobial activity against Staphylococcus aureus. In these sets of non-nucleoside fused heterocyclic compounds electron withdrawing groups at position 5 of the benzazoles increased the activity against C. albicans.