微量稀释法检测申克孢子丝菌酵母相体外抗真菌药物敏感性

目的用液基微量稀释法观察双相真菌申克孢子丝菌酵母相体外抗真菌药物敏感性。方法将54株申克孢子丝菌临床株于脑心浸液琼脂培养基连续传代获得酵母相,参考美国临床实验室标准化委员会(CLSI)的微量稀释法M27-A2检测菌株酵母相对碘化钾、氟康唑、伊曲康唑和特比萘芬的体外敏感性,并观察碘化钾对伊曲康唑和特比萘芬体外抑菌作用的影响。质控株为克柔念珠菌ATCC6258。结果碘化钾体外无抑菌作用;氟康唑最小抑菌浓度(MIC)几何均数大于64μg/mL;伊曲康唑和特比萘芬MIC几何均数分别为0.98μg/mL和0.17μg/mL。伊曲康唑及特比萘芬的MIC值分别高于伊曲康唑+碘化钾及特比萘芬+碘化钾(P均<0.05)。来源皮肤固定型的菌株与来源皮肤淋巴管型菌株相比MIC值差异无统计学意义(P>0.05)。结论改良的M27-A2方法适用于检测申克孢子丝菌酵母相体外敏感性;酵母相时碘化钾对伊曲康唑、特比萘芬体外抑菌作用有一定的加强效应。     

应用微量稀释法对伊曲康唑于皮肤癣菌MIC影响因素分析

测定抗真菌药物 MIC的方法和程序急待标准化 ,多种实验因素可影响 MIC值。我们用微量稀释法测定了 2 0株红色毛癣菌对伊曲康唑的敏感性 ,用正交设计表 L9( 34 )法分析比较了不同 p H值、培养温度、接种密度及结果判读时间等对 MIC值的影响。结果显示在 RPMI 1640液基上 ,p H7.0、2 7℃培养、1× 10 5个孢子 /ml接种量以及 8天培养期为适宜实验条件。这些为今后测定丝状真菌对抗真菌药物敏感性实验的标准化打下了基础。     

流式细胞计数用于念珠菌的快速药敏试验研究

目的 探索应用流式细胞计数(FCM)进行念珠菌快速药敏试验.方法 参照M27-A标准化方案,应用FCM检测氟康唑(FCZ)对21株念珠菌的抗菌活性,药物作用时间3h,非渗透性荧光染料碘化丙啶(PI)染色0.5h.根据不同药物浓度作用下各药物浓度管荧光强度的梯度变化趋势,将平均荧光均道值大于生长对照管60%的最小药物浓度设定为最小抑菌浓度,即MIC值.并将结果与M27-A标准化方案进行比较.结果 FCM所测MIC值范围在0.125~64μg/mL之间,M27-A标准化方案所测MIC值在0.25~64μg/mL之间,统计学检验两者差异无显著性(P>0.05),结果有很好的一致性.FCM试验全过程仅需4h左右.结论 FCM药敏试验具有快速、敏感、可靠的优势.     

肉桂醛体外抗真菌作用初探

本文测定肉桂醛对部分常见真菌的体外抗菌谱.发现其对8种酵母及酵母样真菌的MIC值范围为1.95-62.51μg/ml,以克柔假丝酵母和热带假丝酵母最敏感;对皮癣菌及深部真菌的M工C范围为7.82-125.00μg/ml,以紫色癣菌最敏感.同时作了药物对10株白念菌的时间抑菌动态观察,抑菌曲线表明孵育8小时时,出现抑菌最低点.采用酮康唑对照,发现该药的MIC值普遍较酮康唑高.扫描和透射电镜也证实该药对白念菌、石膏毛癣菌和申克氏孢子丝菌有较强破坏作用,见菌外形改变及细胞壁破损.     

茶皂甙体外抗真菌作用和治疗皮肤癣菌病的疗效观察

本文报告茶皂甙的体外抗真菌作用和茶皂甙酊治疗体癣、股癣的疗效观察。体外试验显示茶皂甙有较强的抑制真菌作用,对红色癣菌、石膏样癣菌、断发癣菌、黄癣菌、紫色癣菌、絮状表皮癣菌的MIC为0.125～1g/L,对白念珠菌为0.25～0.0625g/L,应用茶皂甙酊治疗皮肤癣菌病的结果是十分有效的。     

念珠地丝菌对五种抗真菌药的体外敏感性及电镜观察

目的 探讨5种临床常见抗真菌药对念珠地丝菌的体外抑制作用,并观察用药后念珠地丝菌的形态学变化。方法 5种临床常见抗真菌药为特比萘芬、两性霉素B、氟胞嘧啶、氟康唑和伊曲康唑。参照美国国家临床试验标准委员会修订的M27?A2方案及相关文献,测定各药物对念珠地丝菌的MIC值。应用扫描电镜观察特比萘芬作用后念珠地丝菌的形态学变化。结果 5种药物对念珠地丝菌的MIC值依次为：特比萘芬0.01 μg/ml、两性霉素B 0.4 μg/ml、氟胞嘧啶2 μg/ml、氟康唑2.69 μg/ml、伊曲康唑 0.25 μg/ml。念珠地丝菌对特比萘芬、两性霉素B、氟胞嘧啶、氟康唑均敏感,对伊曲康唑为剂量依赖性敏感。扫描电镜可见,经特比萘芬作用后念珠地丝菌菌体表面出现粗糙、皱缩、不规则缺损及孔洞,甚至呈碎片状。结论 特比萘芬对念珠地丝菌的抗菌活性最显著,在一定范围内,特比萘芬浓度越高,念珠地丝菌被破坏的程度越严重。     

In vitro susceptibility of Trichomonas vaginalis to metronidazole and treatment outcome in vaginal trichomoniasis

We have identified Trichomonas vaginalis strains resistant in vitro to metronidazole, especially under aerobic conditions. Since little is known about the relationship of treatment outcome to metronidazole susceptibility of T. vaginalis, we studied the aerobic and anaerobic susceptibility to metronidazole of 310 clinical isolates of T. vaginalis. Of 199 patients with known outcomes after metronidazole treatment for vaginal trichomoniasis, the geometric mean minimal lethal concentration (MLC) under aerobic conditions for trichomonads associated with cases cured by a single 2-g dose was 24.1 micrograms/ml (n = 146), while that of treatment-resistant isolates (n = 53) was 195.5 micrograms/ml. The corresponding mean anaerobic MLC values were 1.6 and 5.05 micrograms/ml, respectively. The average aerobic:anaerobic MLC ratio was about twofold higher for the resistant isolates. Treatment resistance was more frequent at aerobic MLC values of greater than 25 micrograms/ml or anaerobic values of greater than 1.6 micrograms/ml. Although there was overlap of the metronidazole susceptibility distribution of susceptible and resistant isolates, significant resistance to treatment was common when isolates of T. vaginalis had aerobic MLC values of greater than 100 micrograms/ml or anaerobic MLC values of greater than 3.1 micrograms/ml.     

IN VITRO SUSCEPTIBILITY OF DERMATOPHYTES TO ANTIFUNGAL DRUGS: A COMPARISON OF TWO METHODS

Background. There is an evidence that some fungal strains are resistant to certain antimycotic drugs with resulting therapeutic failures. Unlike testing of antibacterial agents, the activity of these drugs against the given fungal strains is not tested routinely. The paper presented is an approach to such a procedure. Methods. The minimum inhibitory concentration (MIC) values obtained using the dilution method were compared with the diameters of growth inhibition zones using the disk-diffusion method. Seven antifungal drugs (graiseofulvin, natamycin, naftifine, clotrimazole, bifonazole, miconazole, ketoconazole) were tested with 80 fungal strains, including Trichophyton rubrum, T. mentagrophytes, Microsporum canis, and Epidermophyton floccosum. Results. The majority of fungal strains were inhibited by griseofulvin at concentrations not exceeding 10 mg/L. The least susceptible was T. mentagrophytes. The most effective drug was naftifine, and the least effective was natamycin. Naftifine inhibited 98.75% of the strains tested at a concentration not exceeding 1.0 mg/L. An increase of the MIC value was accompanied by a decrease in the growth inhibition diameter. The relation was statistically significant. Conclusions. T. rubrum strains are more susceptible to antifungal drugs than T. mentagrophytes. Naftifine proved to be the most effective drug in dermatophytoses. Natamycin is a useful drug against yeast-like fungi but not against dermatophytes. The disk-diffusion method of fungal susceptibility assessment yields data consistent with results obtained from the dilution method and is recommended for routine use because of its simplicity.     

虫霉病

虫霉病是一种少见的深部真菌病,主要由蛙粪霉(固孢、林和裂孢蛙粪霉)、耳霉(冠状和异孢耳霉)两类致病菌引起,多发生在热带或亚热带,在临床上主要表现为皮肤和皮下组织肿块,质地硬而无触痛；胃肠道蛙粪霉病则有胃肠不适,还可有Crohn病症状及腹部肿块：全身无发热。组织真菌培养可见到宽、有或无隔的菌丝,孢子囊、孢子囊冠及接合孢子。组织病理呈肉芽肿改变,并可见有不规则分枝的菌丝,壁薄偶尔有隔。治疗应用碘化钾、磺胺及其他抗真菌药或联合用药均取得较好的疗效,但疗程比较长。     

葡萄糖消耗试验用于体外抗真菌药物敏感性试验

目的：评价葡萄糖消耗试验应用于体外抗真菌药物敏感性试验的可行性。方法：分析8种抗真菌药物与40株临床分离菌株在仅含葡萄糖为碳源的酵母氮培养基(YNBG)上孵育后葡萄糖的浓度变化，反映真菌细胞的受抑制程度，同时与NCCLS M27-A液基微量稀释法对比。结果：葡萄糖消耗试验在念珠菌和隐球菌的药敏测定中至少可以节省20小时，对须癣毛癣菌则至少可节省40小时；除两种方法测得的联苯苄唑MIC符合程度较低外，两种方法测得的两性霉素B、特比萘芬、伊曲康唑、酮康唑、咪康唑、氟康唑、环吡酮胺的MIC符合程度均较高。在评价由较低或较高浓度药物诱导的真菌细胞受抑制程度时，葡萄糖消耗试验更精确。结论：葡萄糖消耗试验具有客观量化、快捷、精确的优点，具有推广和应用价值。     

In vitro antifungal susceptibility of dermatophyte strains causing tinea pedis and onychomycosis in patients with non‐insulin‐dependent diabetes mellitus: a case‐control study

Background The efficacy of antifungal treatment may be reduced and/or delayed in diabetic patients. To date, no study has investigated the in vitro antifungal susceptibility of dermatophytes in this patient group. Objective We aimed to determine the dermatophyte species causing tinea pedis and onychomycosis, and in vitro susceptibility of these dermatophytes to terbinafine, itraconazole, and fluconazole in patients with non‐insulin‐dependent diabetes mellitus. We compared the findings in diabetic patients with those in non‐diabetic individuals. Materials and methods One hundred patients with non‐insulin‐dependent diabetes mellitus and 100 otherwise healthy controls clinically suspected with tinea pedis and/or onychomycosis were included. Skin scrapings and/or nail clippings were taken and cultured on Sabouraud dextrose agar, mycobiotic agar, and dermatophyte test medium. In vitro antifungal susceptibility tests were carried out according to the Clinical and Laboratory Standards Institute (CLSI) M‐38P protocol with some modifications. Results Fifty‐seven samples of 54 diabetics and 50 samples of 50 controls grew dermatophytes. In both groups, Trichophyton rubrum was the most common isolate. Mean MIC values of terbinafine, itraconazole, and fluconazole for all of the isolated dermatophyte strains were similar in two groups (P > 0.05). The difference in mean MIC values of three antifungals for T. rubrum and T. mentagrophytes between two groups was not statistically significant (P > 0.05). Conclusions Dermatophyte types causing tinea pedis and onychomycosis, their frequency patterns, and in vitro activity of three antifungals against dermatophytes in diabetics are similar to the non‐diabetics. Terbinafine is the most active agent in vitro in both groups.     

孢子丝菌病药敏和治疗进展

孢子丝菌病是一种深部真菌病,其致病菌申克孢子丝菌为一种双相真菌,双相真菌的体外药敏试验至今仍无统一标准.饱和碘化钾溶液、伊曲康唑、特比萘芬为治疗孢子丝菌病的常用药物,临床中选择合适的抗真菌药物对治疗此病具有重要意义.体外药敏试验是评价抗真菌药物活性的重要方法,也是选择药物的依据之一.     

In vitro activity of fluconazole,itraconazole, voriconazole and terbinafine against fungi causing onychomycosis

Background. Onychomycosis is one of the commonest dermatological diseases worldwide. The antifungal activity of current medications varies, and treatment failure occurs in 25–40% of treated patients. Aims. To evaluate the in vitro antifungal activity of itraconazole, fluconazole, terbinafine and voriconazole against isolates taken from patients with onychomycosis. Methods. Nail isolates were evaluated according to methods described in the protocols of the Antifungal Susceptibility Testing Subcommittee of the European Committee on Antimicrobial Susceptibility Testing (AFST‐EUCAST) and the Clinical and Laboratory Standards Institute (CLS M38‐A), and a CLSI M38‐A modified technique for dermatophytes. Antifungal agents tested included terbinafine, itraconazole, voriconazole and fluconazole. Results. In total, 103 isolates of Candida species (n = 58), Fusarium species (n = 10), Fusicoccum dimidiatum (n = 4), Scytalidium hyalinum (n = 1) and dermatophytes (n = 30) were evaluated. Itraconazole and voriconazole were the most active agents against Candida species, whereas terbinafine and voriconazole were most potent against dermatophytes. Fusarium species had the highest minimum inhibitory concentration (MIC) values with all antifungal agents. Conclusions. The aetiological agents of onychomycosis that we found differ from those found in other countries, suggesting that the heat and humidity of the Colombian climate could favour yeast nail infections. The lowest MICs for Candida species (obtained with voriconazole, followed by itraconazole) may be explained by emerging resistant strains. Against dermatophytes, the lowest MICs were obtained with terbinafine, followed by voriconazole. MIC values for the evaluated agents were higher for non‐dermatophyte filamentous fungi than for other fungi. As MIC breakpoints have not yet been established for onychomycosis therapies, it remains unclear if in vitro activities of antifungal drugs are predictive of clinical outcome. Well‐designed clinical studies are necessary to assist clinicians in choosing the best antifungal agents.     

结缔组织病患者咽部定植的白念珠菌毒力测定

目的了解结缔组织病患者咽部定植的白念珠菌芽管生成长度和分泌型天冬氨酸蛋白酶活性。方法采用病例对照方法对病例组30株和健康组4株白念珠菌芽管生成长度和分泌型天冬氨酸蛋白酶活性进行测定。结果病例组菌株芽管长度为(8.425±1.513)μm,显著高于健康组的(6.495±0.375)μm(P<0.05);病例组分泌型酸性蛋白酶活性Pa值为0.490±0.053,低于健康组的0.610±0.056(P<0.01)。白念珠菌生成的芽管长度与其分泌的酸性蛋白酶活性Pa值呈负性相关(r=-0.874,P<0.01)。结论与健康人相比,结缔组织病患者咽部定植的白念珠菌毒力较强,有更强的潜在致病力。     

国产氟康唑抗真菌药效学研究

对国产氟康唑作体内外药效学研究中发现不论在体外或体内均具有较好的广谱抗真菌作用。体外对３１株白念珠菌（白念ＭＩＣ为０．８～２００ｍｇ／Ｌ）、２４株新型隐球菌（新隐ＭＩＣ为２．２４～３５．４ｍｇ／Ｌ）及部份皮癣菌等均有较好的抑菌作用，其ＭＩＣ范围及ＭＩＣ５０、ＭＩＣ９０的结果均与国外同类制品（０．２～＞１００ｍｇ／Ｌ及３．１３～１２．５ｍｇ／Ｌ）相似，且对国外尚未报道的马尔尼菲青霉菌均有一定抑菌作用。体内对白念与新型隐球菌脑膜炎的５０％致死保护量（ＥＤ５０）为０．０５７～０．９４９ｍｇ／ｋｇ，与国外同类产品相当或更佳（０．０８～１．２５ｍｇ／ｋｇ）。提示国产氟康唑与进口制品相似，可用于人类的系统性真菌感染，尤其是对难治的新型隐球菌脑膜炎可获得较好疗效。     

盐酸小檗碱抗白念珠菌体外药敏实验研究

目的 探讨盐酸小檗碱( berberine,BBR)对白念珠菌的体外抗真菌活性。方法 采用美国临床实验室标准化委员会(现称美国临床实验室标准化研究所,CLSI)制订的M27 -A方案中的酵母菌微量稀释法,测定BBR、氟康唑、BBR联合氟康唑对30株临床分离白念珠菌的抗菌活性。结果 盐酸小檗碱对白念珠菌MIC值的几何均数(G)为248.18 mg/L。与320、160、80 mg/L BBR联合后,氟康唑MIC值的几何均数从2.64 mg/L降为0.03、0.35、1.35 mg/L。两药对26.7％的白念菌株有协同抗菌作用,其中包括3株耐氟康唑菌,对56.7％的菌株表现为相加作用,对16.7％的菌株为无关作用,未观察到拮抗作用。结论 BBR在体外对白念珠菌具有抗真菌作用并能增加其对氟康唑的敏感性。     

Inhibition of elastase activity by essential oils in vitro

Summary Background Essential oils are widely used, for example in aromatherapy and aroma massage. In aroma massage, essential oil, diluted with vegetable oil, is rubbed onto the skin. Components of essential oil penetrate into the skin and have an influence on the dermis. Elastase is an enzyme which degenerates dermal elastin. Elastase activity is believed to contribute to cutaneous wrinkling and ageing. Aim To investigate the inhibitory effect of essential oils on elastase activity. Methods Inhibition of elastase activity by various essential oils was assessed using two elastase enzymes: porcine pancreatic elastase (PPE) and human neutrophil elastase (HNE). Results Elastase activity was inhibited by various essential oils, especially by those oils derived from lemons, juniper and grapefruit. Although the specific inhibitory component was not determined, lemon oil had the greatest inhibitory effect on PPE. Some essential oils also inhibited HNE. Conclusions These studies demonstrate a possible rationale for the use of essential oil massage as a preventive treatment for cutaneous wrinkling and ageing.     

伏立康唑与其他5种抗真菌药体外抗念珠菌活性的比较研究

目的比较伏立康唑与其他5种抗真菌药在体外抗深部致病念珠菌的活性。方法按NCCLS推荐的M27A方案(肉汤微量稀释法)检测伏立康唑等6种抗真菌药对6种共52株深部致病念珠菌临床分离株在体外的抗真菌活性。结果对受试菌株总体而言,伏立康唑的活性最高(MIC90≤0.5μg/ml)。伏立康唑对6种念珠菌的MIC90从小到大依次为白念珠菌、热带念珠菌、近平滑念珠菌、克柔念珠菌、光滑念珠菌和法氏念珠菌。伏立康唑对各受试菌种的MIC90均低于氟康唑和特比萘芬;对白念珠菌、热带念珠菌、光滑念珠菌和近平滑念珠菌的MIC90低于伊曲康唑;对克柔念珠菌和法氏念珠菌的MIC90低于两性霉素B。结论伏立康唑在体外能有效抑制深部致病念珠菌的生长,其抗菌谱较广,尤其对唑类药耐药的两种念珠菌——克柔念珠菌和光滑念珠菌也具有较好的抗真菌活性,部分菌种抗真菌活性甚至优于氟康唑和伊曲康唑。     

盐酸小檗碱联合抗真菌药物对红色毛癣菌的体外药敏实验

目的观察盐酸小檗碱联合不同浓度抗真菌药物对红色毛癣菌的体外抗菌活性。方法参照美国临床实验室标准化研究所(CLSI)颁布的M38-A2方案和相关文献对临床分离的30株红色毛癣菌进行体外药敏实验。其中盐酸小檗碱的MIC值定义为与生长对照相比,肉眼观察出现80%生长抑制的最低药物浓度。采用分数抑菌浓度(FIC)评定两药联合效果。FIC≤0.5,两药为协同作用;0.54,两药为拮抗作用。结果盐酸小檗碱和不同浓度伊曲康唑联合,主要为无关作用。盐酸小檗碱和不同浓度特比奈芬、伏立康唑联合,均为拮抗作用。16.0μg/mL氟康唑组与盐酸小檗碱联合时FIC≤0.5所占比例为83.33%,协同作用最强;2.0～8.0μg/mL卡泊芬净组与盐酸小檗碱联合时,FIC≤0.5所占比例最高,均为63.33%,协同作用最强,差异均有统计学意义(P均<0.05)。结论盐酸小檗碱与高浓度氟康唑、卡泊芬净联合,体外对红色毛癣菌有较强的协同作用。     

女性痤疮患者卵泡期血清六项性激素水平的测定

目的探讨女性寻常痤疮患者体内性激素水平的改变。方法采用电化学发光免疫分析法对30例青春期女性痤疮患者及20例迟发性女性痤疮患者卵泡期血清六项性激素水平进行检测,并分别与相应年龄段的正常女性各15人作对照。结果女性青春期痤疮患者血清雌二醇水平明显低于同龄正常对照组(P<0.05);睾酮/雌二醇比值明显高于正常对照组(P<0.01);促卵泡素明显高于正常对照组(P<0.05);女性迟发性痤疮患者血清睾酮水平较相应年龄的正常对照组显著升高(P<0.05)。结论青春期女性痤疮发病的主要原因可能是由于雌激素分泌不足,使血清内睾酮水平相对增多所致。女性迟发性痤疮发病的主要原因则可能与雄激素分泌增多有关。