应用CLSI M38-A2方案测定须癣毛癣菌对抗真菌药物敏感性

目的:对我国代表地区须癣毛癣菌临床分离株作抗真菌药物敏感性测定,进一步验证CLSI的M38-A2方案.方法:选取我国南北方8个省市地区经表型和分子生物学鉴定的趾间型毛癣菌38株和苯海姆节皮菌6株,采用M38-A2方案测定氟康唑、伊曲康唑、伏立康唑、特比萘芬、灰黄霉素、联苯苄唑、环吡酮胺和阿莫罗芬等8种常见抗真菌药物的最小抑菌浓度(minimal inhibitory concentration,MIC).结果:氟康唑、伊曲康唑、伏立康唑、特比萘芬、灰黄霉素、联苯苄唑、环吡酮胺和阿莫罗芬对趾间型毛癣菌株的MIC值(μg/mL)范围分别为0.25-32、0.0312-1、0.0156-0.0625、0.000937-0.00781、0.0625-1、0.0312-2、1-2、0.00781-0.0625;对苯海姆节皮菌株的MIC值(μg/mL)范围分别为≥64、2、0.25-0.5、0.000937-0.00381、1、2-4、1-2、0.0312-0.0625.不同抗真菌药物对趾间型毛癣菌及苯海姆节皮菌的药敏有明显差别(P<0.001);趾间型毛癣菌和苯海姆节皮菌对伊曲康唑、灰黄霉素、环吡酮胺、伏立康唑和氟康唑的药敏差异有统计学意义,对特比萘芬、阿莫罗芬和联苯苄唑的药敏差异无统计学意义.结论:趾间型毛癣菌和苯海姆节皮菌之间对伊曲康唑、灰黄霉素、环吡酮胺、伏立康唑和氟康唑的药敏有明显差异.M38-A2方案有较好的重复性和稳定性,适合用来体外测定须癣毛癣菌对抗真菌药物的敏感性.     

外用和口服抗真菌药治疗足癣疗效评价

目的分别评价外用和口服抗真菌药治疗足癣的疗效,为足癣治疗提供参考。方法收集国内外已公开发表的有关外用和口服抗真菌药治疗足癣的临床随机对照试验进行分析评价。结果共有24篇文献入选,总病例数为3059例。外用联苯苄唑乳膏的疗效优于咪康唑和克霉唑乳膏;特比萘芬的疗效明显优于克霉唑乳膏;缺乏特比萘芬和联苯苄唑乳膏疗效比较的相关文献。口服特比萘芬(250mg/d,治疗2周)或伊曲康唑(100mg/d,治疗4周)均明显优于安慰剂和灰黄霉素(500mg/d,治疗4周或6周)。结论外用特比萘芬和联苯苄唑乳膏疗效均较理想;口服特比萘芬和伊曲康唑均有较好的临床疗效。     

1％联苯苄唑霜治疗足癣长期疗效观察

联苯苄唑（Bifonazole）属于咪唑类广谱抗真菌药物。对皮肤癣菌、酵母菌、非皮肤癣菌性霉菌具有较高的抗菌活性;对革兰阳性球菌和棒状杆菌亦有效;还有明显的抗炎作用,其抗炎作用相当于1％氢化可的松霜。本研究采用多中心、随机开放、平行对照研究评价由拜耳医药保健有限公司生产的1％联苯苄唑霜（美克）治疗足癣的长期疗效,并与2％硝酸咪康唑霜（达克宁）和1％特比萘芬霜（丁克）进行对照。     

1%盐酸布替萘芬溶液治疗趾间型足癣的疗效和安全性

本研究评价以1％盐酸布替萘芬（butenafine hydrochloride）溶液治疗趾间型足癣的疗效和安全性,并与市售1％联苯苄唑溶液的作用进行比较。     

萘替芬酮康唑乳膏治疗皮肤真菌病安全性与有效性的meta分析

目的综合评价萘替芬酮康唑乳膏治疗皮肤真菌病的有效性及安全性。方法在The Cochrane Library、Medline、Embase、CBM、中国知网(CNKI)、维普和万方数据库上检索萘替芬酮康唑乳膏与其他抗真菌药物单药治疗皮肤真菌病的随机对照研究,用NoteExpress进行文献筛选及资料提取,使用RevMan5.3进行系统评价。结果最终纳入43篇文献,共5413名患者,治疗有效率为85.27%。分析结果显示萘替芬酮康唑乳膏疗效优于硝酸咪康唑[RR=1.32,95%CI (1.26,1.38)],优于联苯苄唑治疗4周组[RR=1.31,95%CI (1.12,1.54)],优于酮康唑[RR=1.25,95%CI (1.14,1.36)]。萘替芬酮康唑在复发率及不良反应发生率上低于其他药物。结论萘替芬酮康唑乳膏可有效治疗皮肤真菌病,且更加安全。     

低浓度他克莫司与抗真菌药物联合的体外抗真菌效应

目的测定他克莫司(FK-5066)与抗真菌药物对浅部致病真菌的联合效应。方法参考CLSI的M27-A3和M38-A2方案分别测定酮康唑对念珠菌、特比萘芬对皮肤癣菌的最低抑菌浓度(MIC);利用棋盘法,测定他克莫司与酮康唑或特比萘芬的联合抗真菌效果。结果酮康唑单独对念珠菌MIC50和MIC90分别为:4μg/m L和8μg/m L;与他克莫司联合应用时,对53.3%的菌株有联合效应(n=15),对其余菌株无协同效应。特比萘芬单独对皮肤癣菌MIC50和MIC90分别为:0.06μg/m L和0.125μg/m L;与他克莫司联合应用时,对46.7%菌株(n=15)有协同效应,对其余菌株表现为无关。结论低浓度他克莫司在体外与抗真菌药物联合应用时,能够降低药物对部分菌株的MIC,未发现拮抗效应。     

硝酸舍他康唑的临床应用研究进展

硝酸舍他康唑(sertaconazole)是一种新型、广谱、高效的外用唑类抗真菌药。本文就其药理作用、抗炎、止痒、抗真菌临床疗效及安全性评价等方面加以综述。新的剂型-微乳凝胶,增强皮肤渗透性和存留时间,而且能减少对皮肤的刺激。外用硝酸舍他康唑治疗皮肤癣菌病和浅表念珠菌感染见效快,治愈率高,疗程短。根据药代动力学,舍他康唑只需每日用药一次,加之其止痒活性,增加了患者依从性,从而提高了临床治愈率。最新临床研究发现外用硝酸舍他康唑乳膏对脂溢性皮炎、特应性皮炎有良好的疗效。     

8种抗真菌药物对浅表真菌病致病菌株体外敏感性检测

目的确定8种抗真菌药物对100株浅表致病真菌的体外药物敏感性。方法参考美国临床和实验室标准化研究所(Clinical and Laboratory Standards Institute,CLSI)微量液基稀释法M27-A3和M38-A2方案,采用8种抗真菌药物对100株常见浅表致病真菌进行体外药物敏感性测定。结果体外药敏实验结果显示,8种药物对皮肤癣菌表现出良好的体外药物敏感性,其中特比萘芬(GM MIC,0.011μg/mL,MIC范围,0.004~0.06μg/mL)具有最强的抗真菌活性。咪康唑(GM MIC,0.147μg/mL,MIC范围0.015~1μg/mL)具有最强的抗念珠菌活性。酮康唑(GM MIC,0.097μg/mL,MIC范围0.03~1μg/mL)则对马拉色菌具有最佳体外抗真菌效果。结论体外药敏实验结果表明常见浅表致病真菌对8种常用抗真菌药物表现出不同的敏感性。酮康唑具有广谱高效的特点,对临床常见皮肤癣菌、念珠菌、马拉色菌均具有良好的体外抗真菌活性。特比萘芬和萘替芬对皮肤癣菌具有最佳的抗真菌活性。     

球形孢子丝菌致婴儿固定型孢子丝菌病1例及相关实验研究

报道1例由球形孢子丝菌所致的婴儿固定型孢子丝菌病。患儿女,3个月,因左眼下内侧皮损2个月就诊,皮损脓液标本进行真菌培养,对培养获得菌株进行形态学、生理学和分子生物学鉴定,并进行药物敏感性检测。真菌培养阳性,镜下可见典型的套袖样菌丝。钙调蛋白基因序列分析鉴定为球形孢子丝菌。药敏试验显示特比萘芬和伊曲康唑对该菌株的菌丝相最低抑菌浓度（minimal inhibitorycon centration,MIC）分别为0．5μg／mL和0．5μg／mL;对该菌株的酵母相的MIC值分别为0．25μg／mL和0．5μg／mL。给予患者口服特比萘芬32．5mg／d治疗10周后皮损消退呈瘢痕化修复。依据临床及实验室检查确诊该病例为球形孢子丝菌所致固定型孢子丝菌病,特比萘芬治疗本病例显示较好疗效。     

85株皮肤癣菌感染的病原菌分析及标准化体外药物敏感性研究

目的分析深圳地区教学医院门诊皮肤癣菌感染的病原菌特点,测定7种抗真菌药物对85株皮肤癣菌的体外抗真菌活性。方法收集疑诊为皮肤癣菌感染的患者的皮屑(或甲板刮屑、毛发)进行KOH直接镜检和培养。并通过测定皮肤癣菌临床分离菌株的核糖体DNA内转录间隔区(ITS)区序列鉴定菌种。参考CLSI M38-A2方案,测定7种抗真菌药物对85株鉴定为皮肤癣菌的临床分离菌株的体外抗真菌活性。结果 161例疑诊为皮肤癣菌感染的患者中KOH直接镜检皮肤癣菌的阳性率为67.7%,真菌培养皮肤癣菌的阳性率为52.8%。菌种鉴定结果为红色毛癣菌68例、犬小孢子菌7例、石膏样小孢子菌3例、趾间毛癣菌5例、紫色毛癣菌1例、断发毛癣菌1例。体外药敏试验显示特比萘芬(GM MIC,0.032μg·mL~(-1), MIC范围, 0.001～0.125μg·mL~(-1))对6种皮肤癣菌表现出良好的体外抗真菌活性,而伏立康唑(GM MIC, 0.041μg·mL~(-1), MIC范围, 0.032～0.125μg·mL~(-1))在唑类药物中的体外MIC值相对较低。结论本研究中皮肤癣菌感染的致病菌以红色毛癣菌为主。标准化药敏试验证实特比萘芬和伏立康唑对分离的皮肤癣菌具有较强体外抗菌活力,适用于本地区皮肤癣菌感染的治疗。     

美浮特皮肤抗菌液对致足癣真菌抗真菌后效应的体外研究

目的：评价美浮特^R皮肤抗菌液对致足癣真菌皮肤癣菌和白念珠菌的体外抗真菌活性及抗真菌后效应。方法采用美国 CLSI M27-A3和 M38-A2方案测定美浮特皮肤抗菌液对足癣常见致病真菌的最低抑菌浓度（ MIC）;并以白念珠菌（ATCC90028）为指示菌测定美浮特皮肤抗菌液测定时间-杀菌曲线,同时测定其对白念珠菌的抗真菌后效应（post-antifungal effect,PAFE）。结果美浮特皮肤抗菌液对4属6种57株 MIC 的范围为1：40-1：160、MIC50为1：80、MIC90为1：40;对白念珠菌的 MIC 范围为1：40-1：80、对皮肤癣菌的 MIC 范围为1：40-1：160。该抗菌液具有很强的杀菌作用,且随着药物浓度的降低,杀菌速度和程度随之变化。该抗菌液对白念珠菌0.5MIC、MIC、2MIC 的 PAFE 分别为0.85 h、2.1 h、3.59 h;且 PAFE 时间的延长与药物浓度呈正相关。结论美浮特^R皮肤抗菌液对致病真菌皮肤癣菌、白念珠菌具有快速、有效、持续的杀菌作用,该抗菌液对皮肤癣菌较白念珠菌具有更强的抗真菌作用。且该抗菌液对白念珠菌具有较长时间的后效应,可以广泛应用于临床治疗皮肤癣菌及白念珠菌所致的感染。     

In vitro susceptibility of dermatomycoses agents to six antifungal drugs and evaluation by fractional inhibitory concentration index of combined effects of amorolfine and itraconazole in dermatophytes

To investigate the antifungal drug susceptibility of fungi responsible for dermatomycoses, minimum inhibition concentration (MIC) tests were performed in 44 strains of dermatophytes, including Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton verrucosum, Trichophyton tonsurans, Microsporum canis, Microsporum gypseum and Epidermophyton floccosum, with six antifungal drugs (amorolfine, terbinafine, butenafine, ketoconazole, itraconazole and bifonazole) by broth microdilution assay according to Clinical Laboratory Standard Institute protocols. Six possible dermatomycosis‐causing non‐dermatophytic fungi were also tested. The two major causes of tinea, T. rubrum and T. mentagrophytes, showed significantly different sensitivities to ketoconazole and bifonazole. Clinically derived dermatophytes were sensitive to the six antifungal drugs tested. However, non‐dermatophytes, especially Fusarium spp., tended to be resistant to these antifungal drugs. In Trichophyton spp., the MICs of non‐azole drugs had narrower distributions than those of azoles. To evaluate the effects of antifungal drug combinations, the fractional inhibitory concentration index was calculated for the combination of amorolfine and itraconazole as representative external and internal drugs for dermatophytes. It was found that this combination had synergistic or additive effects on most dermatophytes, and had no antagonistic effects. The variation in susceptibility of clinically derived fungal isolates indicates that identification of causative fungi is indispensable for appropriately choosing effective antifungal drugs in the early stages of infection. The results of combination assay suggest that multiple drugs with different antifungal mechanisms against growth of dermatophytes should be used to treat refractory dermatomycoses, especially onychomycosis.     

覆盆子提取物联合唑类药物抗真菌活性研究

目的 探讨中药覆盆子提取物联合唑类药物的体外抗真菌作用.方法 采用CLSI公布的M27-A方案微量液基稀释法和棋盘式微量稀释法,测定覆盆子提取物单用及联合唑类药物对不同念珠菌的MIC值和FICI指数.结果 覆盆子不同溶液提取物与氟康唑均表现出协同关系,以覆盆子醇提物为例,单用对念珠菌的MIC80测定值范围主要集中在0.16～1.25 mg/mL,与氟康唑合用后表现出协同关系(FICI≤0.5),且MIC80测定值范围降至0.01 ～0.04 mg/mL;合用后的氟康唑抗真菌活性也明显增强.另外,覆盆子醇提物与不同唑类药物合用后均有协同关系,其MIC80测定值由单用时大于10 mg/mL降至0.04 mg/mL.结论 覆盆子醇提物和唑类药物单用时对耐药念珠菌的抑菌作用较弱,但二者合用后表现出明显的协同关系,对耐药念珠菌的抑菌作用明显增强.     

Synthesis and activities of naphthalimide azoles as a new type of antibacterial and antifungal agents

Naphthalimide-derived azoles as a new type of antimicrobial agents were synthesized and evaluated for their efficiency in vitro against eight bacteria and two fungi by two fold serial dilution technique. Most title compounds exhibited good antimicrobial potency with low MIC values ranging from 1 to 16 μg/mL. Notably, some synthesized compounds displayed comparable or even better antibacterial and antifungal activities against some tested strains than the reference drugs Orbifloxacin, Chloromycin and Fluconazole, respectively.     

In-vitro studies with four new antifungal agents: BAY n 7133, bifonazole (BAY h 4502), ICI 153,066 and Ro 14-4767/002

Four new antifungal agents were compared in vitro with miconazole and ketoconazole. The agents were BAY n 7133 and ICI 153,066, two orally active triazoles, and bifonazole (BAY h 4502) and Ro 14-4767/002, both topical agents. While all four were found to be broad spectrum antifungal agents they also demonstrated certain gaps in their spectra. In general, Ro 14-4767/002 was the most active agent tested whereas bifonazole and BAY n 7133 were the least active. Noteworthy activities included that of Ro 14-4767/002 against Candida albicans, the dermatophytes and Sporothrix schenckii and that of ICI 153,066 against Torulopsis glabrata.     

Elucidation of antifungal metabolites produced by Pseudomonas aurantiaca IB5-10 with broad-spectrum antifungal activity

Antifungal metabolites were isolated from a culture of Pseudomonas aurantiaca IB5-10. Chemical structures of the metabolites were elucidated as phenazine-1-carboxylic acid (PCA; 1), 2-hydroxyphenazine (2-OH-PHZ; 2), and cyclo-(L-Pro-L-Val; 3), respectively, based on spectroscopic methods. Among them, 3 was isolated for the first time from this strain. The antifungal activities of 1-3 were evaluated against a variety of plant pathogens. To the best of our knowledge, the antifungal activities of 3 against plant fungal pathogens have been evaluated for the first time in this work. PCA (1) showed the most potent antifungal activities against Phytophthora capsici, Rhizoctonia solani AG-1(IA), and Pythium ultimum with MICs (microgram/ml) of less than 1.0, 1.3, and 2.0, respectively. On the other hand, 2-OH-PHZ (2) showed potent antifungal activity against R. solani AG-1(IA) with the MIC (microgram/ml) of 2.0, whereas it showed moderate antifungal activity against P. ultimum with the MIC (microgram/ml) of 50.0. In addition, 3 showed antifungal activity against only R. solani AG- 1(IA).     

Carboxylic acid and phosphate ester derivatives of fluconazole: synthesis and antifungal activities

Two classes of fluconazole derivatives, (a) carboxylic acid esters and (b) fatty alcohol and carbohydrate phosphate esters, were synthesized and evaluated in vitro against Cryptococcus neoformans, Candida albicans, and Aspergillus niger. All carboxylic acid ester derivatives of fluconazole (1a-l), such as O-2-bromooctanoylfluconazole (1g, MIC=111 microg/mL) and O-11-bromoundecanoylfluconazole (1j, MIC=198 microg/mL), exhibited higher antifungal activity than fluconazole (MIC > or = 4444 microg/mL) against C. albicans ATCC 14053 in SDB medium. Several fatty alcohol phosphate triester derivatives of fluconazole, such as 2a, 2b, 2f, 2g, and 2h, exhibited enhanced antifungal activities against C. albicans and/or A. niger compared to fluconazole in SDB medium. For example, 2-cyanoethyl-omega-undecylenyl fluconazole phosphate (2b) with MIC value of 122 microg/mL had at least 36 times greater antifungal activity than fluconazole against C. albicans in SDB medium. Methyl-undecanyl fluconazole phosphate (2f) with a MIC value of 190 microg/mL was at least 3-fold more potent than fluconazole against A. niger ATCC 16404. All compounds had higher estimated lipophilicity and dermal permeability than those for fluconazole. These results demonstrate the potential of these antifungal agents for further development as sustained-release topical antifungal chemotherapeutic agents.     

Ibuprofen, a unique anti-inflammatory compound with antifungal activity against dermatophytes

Ibuprofen showed significant antifungal activity in vitro against dermatophytes at pH 5 (MIC: 5–40 μg ml^-1). In this respect it is comparatively more efficient than two well known and medically used antifungal compounds, benzoic and salicylic acids. This compound with anti-inflammatory activity which is not found in any other conventional antifungal organic acids, may have clinical prospects.     

Antifungal activities of the essential oils in Syzygium aromaticum (L.) Merr. Et Perry and Leptospermum petersonii Bailey and their constituents against various dermatophytes

This study was carried out in order to investigate the potential of using plant oils derived from Leptospermum petersonii Bailey and Syzygium aromaticum L. Merr. Et Perry as natural antifungal agents. The antifungal effects of essential oils at concentrations of 0.05, 0.1, 0.15, and 0.2 mg/ml on the dermatophytes Microsporum canis (KCTC 6591), Trichophyton mentagrophytes (KCTC 6077), Trichophyton rubrum (KCCM 60443), Epidermophyton floccosum (KCCM 11667), and Microsporum gypseum were evaluated using the agar diffusion method. The major constituents of the active fraction against the dermatophytes were identified by gas chromatography-mass spectrometry and high-performance liquid chromatography analysis. The antifungal activities of S. aromaticum oil (clove oil) against the dermatophytes tested were highest at a concentration of 0.2 mg/ml, with an effectiveness of more than 60%. Hyphal growth was completely inhibited in T. mentagrophytes, T. rubrum, and M. gypseum by treatment with clove oil at a concentration of 0.2 mg/ml. Eugenol was the most effective antifungal constituent of clove oil against the dermatophytes T. mentagrophytes and M. canis. Morphological changes in the hyphae of T. mentagrophytes, such as damage to the cell wall and cell membrane and the expansion of the endoplasmic reticulum, after treatment with 0.11 mg/ml eugenol were observed by transmission electron microscopy (TEM). At a concentration of 0.2 mg/ml, L. petersonii oil (LPO) was more than 90% effective against all of the dermatophytes tested, with the exception of T. rubrum. Geranial was determined to be the most active antifungal constituent of L. petersonii oil. Taken together, the results of this study demonstrate that clove and tea tree oils exhibited significant antifungal activities against the dermatophytes tested in this study.     

中药提取物对酵母菌抗真菌活性研究

目的探讨6味中药2种方法提取成分对酵母菌的抑菌和杀菌作用。方法采用药基琼脂稀释法,测定6味中药水提和醇提成分对白念珠菌和糠秕马拉色菌的MIC和MFC。结果对白念珠菌：水提黄连、醇提黄柏、醇提土槿皮MIC范围分别为0．625—1．25mg／mL、0．625～1．25mg/mL、0．313—0．625mg／mL;均值均为0．625mg／mL;对糠秕马拉色菌：水提和醇提黄连MIC范围分别为0．625～1．25mg／mL和1．25mg／mL,均值均为1．25mg／mL。对白念珠菌：醇提土槿皮MFC范围0．625～2．5mg/mL,均值0．625rag／mL。结论水提黄连、醇提黄柏和土槿皮对白念珠菌有较强抑菌作用,其中醇提土槿皮有较强杀菌作用。水提和醇提黄连对糠秕马拉色菌有较强抑菌作用。