国内多中心甲真菌病病原菌流行病学调查

目的了解国内甲真菌病病原菌的种类和构成情况,掌握流行病学资料。方法对真菌镜检阳性的1 428例甲真菌病患者进行真菌培养和临床分析。结果真菌培养阳性率为53.9%,分离出病原菌800株。其中皮肤癣菌占84.0%,以红色毛癣菌为主(80.9%),其次为指(趾)间毛癣菌和絮状表皮癣菌。酵母菌占11.4%,以念珠菌属为主(10.1%),尤以近平滑念珠菌为主,其次为白念珠菌和热带念珠菌。其他霉菌占4.6%,以枝顶孢霉为主(2.3%),其次曲霉属、青霉属、毛壳菌属、镰刀菌属和帚霉属等。结论本研究显示甲真菌病病原菌以皮肤癣菌为主,其次为酵母菌和霉菌。     

真菌性肉芽肿的研究进展

真菌性肉芽肿在感染性肉芽肿中占第二位。引起肉芽肿的真菌种类繁多,包括球孢子菌、组织孢浆菌、芽生菌、隐球菌、副球孢子菌、孢子丝菌、着色芽生菌、暗色丝孢菌、链格孢霉、霉菌性足分支菌、洛博芽生菌、鼻孢子菌、结合菌、斑替支孢霉等,此外红色毛癣菌、须癣毛癣菌等皮肤癣菌和白念珠菌等也可引起肉芽肿。该文综述真菌性肉芽肿的病因及发病机制、临床表现、诊断及鉴别诊断、治疗等研究进展。     

参照CLSIM38-A方案测定皮肤癣菌临床株对4种抗真菌药物的敏感性

目的测定并比较临床分离的皮肤癣菌对4种常用抗真菌药物的敏感性,探讨CLSIM38-A方案用于皮肤癣菌药敏试验的可行性。方法实验菌株为31株临床近期分离的皮肤癣菌。其中红色毛癣菌14株,须癣毛癣菌14株,犬小孢子菌1株,铁锈色小孢子菌1株,絮状表皮癣菌1株。4种抗真菌药物为益康唑、伊曲康唑、特比萘芬、伏立康唑。采用M38-A方案微量稀释法,并适当调整试验参数进行体外抗真菌药物敏感性试验。结果红色毛癣菌和须癣毛癣菌对以上4种药物的敏感性无明显差异。犬小孢子菌、铁锈色小孢子菌、絮状表皮癣菌对所有4种抗真菌药物的敏感性都低于红色毛癣菌和须癣毛癣菌。结论M38-A方案经过调整适用于皮肤癣菌药敏试验。     

致病真菌对甲侵袭性的体外初步研究

目的观察不同致病真菌在体外对甲板的侵袭能力。方法将分离自甲真菌病患者的致病真菌包括白念珠菌、红色毛癣菌和短帚霉,接种到沙堡培养基中,同时将灭菌甲板埋植入接种处。第28d时,取出甲板,进行病理切片,观察真菌对甲板的侵袭能力。结果病理显示接种于红色毛癣菌及短帚霉的甲板内可见菌丝生长,而接种于白念珠菌的甲板内无菌丝生长。结论皮肤癣菌和霉菌可以在甲板内侵袭生长,而白念珠菌对甲板无侵袭能力。     

红色毛癣菌和枝孢样枝孢霉混合感染导致银屑病患者甲真菌病1例

目的报道1例银屑病患者出现红色毛癣菌和枝孢样枝孢霉混合感染导致的甲真菌病。方法报告病例,对甲标本进行真菌镜检和培养,对病原菌进行形态学及分子生物学鉴定。结果该病例经临床、真菌镜检和真菌培养鉴定,确诊为红色毛癣菌和枝孢样枝孢霉导致的甲真菌病。病原菌通过菌落和显微镜下形态特征结合rRNA内转录间隔区序列分析证实。结论通过形态学及分子生物学鉴定,证实为真菌红色毛癣菌和枝孢样枝孢霉混合感染导致的甲真菌病。     

聚六亚甲基双胍体外抗真菌作用研究

目的探讨聚六亚甲基双胍(PHMB)对浅部真菌病常见致病菌的杀灭作用。方法参照标准的微量液基稀释法和琼脂稀释法检测PHMB粉剂和溶液对红色毛癣菌、须癣毛癣菌、犬小孢子菌及白念珠菌临床分离株的体外最低抑菌浓度(MIC)和最低杀菌浓度(MFC)。结果 PHMB对红色毛癣菌、须癣毛癣菌、犬小孢子菌和白念珠菌临床分离株的MIC范围分别为1.0～4.0μg/mL、2.0～4.0μg/mL、2.0～4.0μg/mL和0.3～5.0μg/mL;MFC范围分别为2.0～8.0μg/mL、 4.0μg/mL、2.0～4.0μg/mL和5.0～10.0μg/mL。琼脂稀释法结果表明PHMB溶液浓度为1.25%时,红色毛癣菌、犬小孢子菌和须癣毛癣菌无菌落生长;PHMB溶液浓度为0.32%时,白念珠菌无菌落生长。加入透皮剂氮酮后,杀菌效果并无改变。结论 PHMB对浅部真菌病常见的致病真菌皮肤癣菌及白念珠菌具有较好的抑制和杀灭作用。     

建立真菌透甲模型及红色毛癣菌和短帚霉透甲时间的比较

目的通过建立真菌透甲模型,观察真菌透甲时间,来探讨红色毛癣菌与短帚霉的相互作用。方法建立真菌透甲模型,分为红色毛癣菌组、短帚霉组、短帚霉-红色毛癣菌混合组,分析各组透甲时间。结果透甲模型中,红色毛癣菌的透甲时间为（9.46±1.89）d,短帚霉的透甲时间为（2.62±0.96）d,混合组中短帚霉的透甲时间为（2.54±0.78）d。结论体外透甲模型中,短帚霉比红色毛癣菌先穿透甲板。混合组中,红色毛癣菌对短帚霉的透甲时间无明显影响。     

不同地区浅部真菌病病原菌流行状况比较

目的了解近半年来华东、华南、华北地区浅部真菌病病原菌的种类、构成情况以及菌种变迁。方法以近半年来在上海、广州、北京3家教学医院皮肤科就诊的浅部真菌病患者为3个地区的代表,对具有典型浅部真菌病临床表现且真菌镜检阳性的患者进行致病真菌的分离培养。结果共分离出致病真菌926株,其中红色毛癣菌544株（58.7%）,念珠菌132株（14.3%）,犬小孢子菌119株（12.9%）,须癣毛癣菌23株（2.5%）,石膏小孢子菌16株（1.7%）。华东地区浅部真菌构成比复杂,华南及华北地区相对简单。结论不同地区浅部致病真菌菌种及构成比存在差异;三地红色毛癣菌均仍占优势;而念珠菌和犬小孢子菌所占比例有上升趋势。     

皮肤癣菌体外蛋白水解酶活性测定

目的观察皮肤癣菌的体外蛋白水解酶活性;比较分离自不同感染部位的红色毛癣菌的体外蛋白水解酶活性。方法实验菌株包括来自不同感染部位的红色毛癣菌22株、须癣毛癣菌3株、犬小孢子菌5株,进行体外培养,并利用9-羟基乙酚噻唑标识的酪蛋白和酶标仪检测真菌细胞外蛋白水解酶的活性。结果须癣毛癣菌的体外蛋白水解酶活性高于红色毛癣菌和犬小孢子菌（P〈0.05）,而红色毛癣菌和犬小孢子菌之间无差异（P〉0.05）。红色毛癣菌的细胞外蛋白水解酶活性在分离自浅部感染部位的菌株之间无差异（P〉0.05）,但高于引起毛癣菌肉芽肿的菌株（P〈0.05）。结论不同的皮肤癣菌体外蛋白水解酶活性可能不同;分离自不同感染部位的同一菌种的体外蛋白水解酶活性也有可能不同。     

卡泊芬净、米卡芬净对8种皮肤癣菌体外抑菌活性的研究

目的评价棘白菌素类抗真菌药物卡泊芬净（caspofungin）、米卡芬净（micafungin）针对常见致病性皮肤癣菌的体外抗菌活性。方法参考CLSI制定的M38-A2方案。测定82株常见皮肤癣菌的最低有效浓度（minimal effective concentrations,MECs）。结果按照MEC90浓度从高到低,米卡芬净对紫色毛癣菌和断发毛癣菌的MEC90是0.25μg/mL;对犬小孢子菌、疣状毛癣菌的MEC90为0.06μg/mL;对红色毛癣菌、须癣毛癣菌、石膏小孢子菌、絮状表皮癣菌的MEC90均在0.03μg/mL。②卡泊芬净对红色毛癣菌、紫色毛癣菌和断发毛癣菌的MEC90为1μg/mL;对须癣毛癣菌、犬小孢子菌、石膏小孢子菌、絮状表皮癣菌和疣状毛癣菌的MEC90为0.5μg/mL。③根据中位数检验,米卡芬净对几种皮肤癣菌的MEC值均低于卡泊芬净的MEC值,统计学比较有显著性差异（P〈0.05）。结论米卡芬净和卡泊芬净对皮肤癣菌有较强的抑菌作用,米卡芬净的MEC值低于卡泊芬净。     

Antifungal Activities of Different Extracts of Marine Macroalgae Against Dermatophytes and Candida Species

Algae are bioactive natural resources, and due to the medical importance of superficial mycoses, we focused the action of macroalgae extracts against dermatophytes and Candida species. Seaweed obtained from the Riacho Doce beach, Alagoas (Brazil), were screened for the antifungal activity, through crude extracts using dichloromethane, chloroform, methanol, ethanol, water and chloroform and hexane fractions of green, brown and red algae in assays with standard strains of the dermatophytes Trichophyton rubrum, T. tonsurans, T. mentagrophytes, Microsporum canis, M. gypseum and yeasts Candida albicans, C. krusei, C. guilliermondi and C. parapsilosis. The M44-A and M27-A2/M38A manuals by CLSI were followed, and the minimum inhibitory concentration (MIC) ranged from 0.03 to 16.00 μg ml(-1), and an inhibition halo of 10.00-25.00 mm was observed for dermatophytes, while for yeast, it was from 8.00 to 16.00 μg ml(-1) and 10.00-15.00 mm. M. canis showed MIC of 0.03 μg ml(-1) and the largest inhibition halo in T. rubrum (25.00 mm) through the use of the methanol extract. For C. albicans, dichloromethane, methanol and ethanol extracts formed the largest inhibition halo. The ethanol extract was shown to be the best inhibiting fungi growth, and chloroform and hexane fractions of H. musciformis inhibited the growth of all dermatophytes and C. albicans, yielding the conclusion that apolar extracts obtained from algae presented the best activity against important pathogenic fungi.     

Antifungal activity of some mediterranean algae

Summary The antifungal activity of 15 mediterranean algae species on some dermatophyte strains (Epidermophyton floccosum, Microsporum canis, M. gypseum and Trichophyton mentagrophytes) and pathogenic yeasts (Candida albicans, C. guillermondii, C. krusei, C. tropicalis and Torulopsis glabrata) has been tested following a modification of Aubert's technique.Among the algae species studied, Falkenbergia rufolanosa is the most active in front of all the fungi tested.     

Occurrence ofCandida strains in cases of paronychia

A total of 43 patients, 11 males and 32 females, with paronychia of the fingernails were examined for the presence of Candida spp. The yeast species isolated were identified using standard laboratory methods, including germ-tube production, morphology on rice agar with Tween 80, and mainly fermentation and assimilation of saccharides. In the male group, two Candida species were detected: C. albicans as the dominant species in 9 patients and C. parapsilosis in 2 cases. Similarly, C. albicans was the prevalent species also in females (n = 17); other Candida species detected were C. parapsilosis (n = 7), C. tropicalis (5) and C. krusei (3). In addition to the genus Candida, the following anaerobic and aerobic microorganisms were isolated from patients of both groups: Fusobacterium spp., Bacteroides spp., Staphylococcus aureus, alpha-hemolytic streptococci, group A beta-hemolytic streptococci, Klebsiella pneumoniae, Neisseria spp. and Pseudomonas aeruginosa.     

Diabetes mellitus and candidiases

Patients in various clinical states of diabetes mellitus (according to the recommendation of the American Diabetes Association) as a primary diagnosis were examined for fungal infections by Candida species. Candida spp. were detected in urine, in the material taken from the mouth cavity, nails, skin lesions, ears and eyes, by cultivation on the Sabouraud agar, CHROMagar Candida, and by saccharide assimilation. In the group of diabetics with symptoms of oral candidiasis and denture stomatitis C. albicans was identified in 8 cases, C. tropicalis in 3, C. parapsilosis in 2; 1 strain of C. guilliermondii was also isolated. In patients with urinary tract infections the presence of C. albicans was shown in 12 cases; C. parapsilosis was detected in 6 cases and two strains of each C. tropicalis and C. krusei were also isolated. In patients with leg ulcers C. albicans (25 cases), C. parapsilosis (5), C. tropicalis (3) and one strain of each C. krusei and C. robusta were isolated. Otomycosis was associated with one strain of C. albicans, C. parapsilosis, C. tropicalis and C. guilliermondii. C. albicans was most frequently associated with onychomycosis, paronychia and endophthalmitis; C. parapsilosis was the second most rated yeast.     

Culture medium alkalinization by dermatophytes

95 dermatophyte strains (12 of Trichophyton mentagrophytes, 12 of T. tonsurans, 11 of T. rubrum, 12 of T. megninii, 12 of T. violaceum, 2 of T. schoenleinii, 1 of T. soudanense, 12 of M. canis, 8 of Microsporum gypseum, 1 of M. ferrugineum and 12 of Epidermphoyton floccosum). 1 of Aspergillus niger, 1 of A. ochraceus, 1 of Paecilomyces sp., 1 of Penicillium sp. and 1 of Candida albicans were grown in Sabouraud liquid medium for the study of pH variation over 6 weeks at room temperature and after 4 weeks at 37 degrees C. All the dermatophyte strains alkalinized the medium. The highest pH values after 2-3 weeks' development in room temperature were produced by T. mentagrophytes, M. gypseum and M. canis, and the lowest by T. rubrum and T. violaceum. After the 4th week the alkalinizing activity became more marked for T. mentagrophytes and remained stationary for T. violaceum, In the 5th week the values produced by T. tonsurans were higher than those for M. gypseum, and those for E. floccosum and T. megninii were higher than those of M. canis. A similar behaviour was observed for T. rubrum and T. megninii and for M. canis, M. gypseum and M. ferrugineum. There seems to be a relation between the alkalinizing capacity and the rapidity and amount of the growth. At 37 degrees C both alkalinization and range of variation of the pH values of the medium became more noticeable for the strains of each species.     

The ability of Candida spp. strains to induce production of tumor necrosis factor and interleukin-6 by whole blood cells

The aim of this study was to investigate the ability of C. albicans, C. glabrata, C. kefyr, C. krusei, and C. parapsilosis to induce production of TNF and IL-6 by whole blood cells of healthy subjects. The highest secretion of TNF and IL-6 was obtained with the 10(7) cells mL(-1) and 10(5) mL(-1) heat killed Candida cells, respectively, for all the species of Candida tested. C. albicans was a better stimulator of TNF and C. glabrata was a better stimulator of IL-6 than other Candida spp. C. krusei was the weakest inductor of IL-6 production.     

口腔黏膜真菌鉴定方法的比较

比较常见用于黏膜真菌菌种鉴别的多种方法,探寻最佳的鉴别方法。采集230例普通人群口腔黏膜样本,分别用玉米吐温-80培养观察厚膜孢子法、糖发酵生化反应法、CHROMagar假丝酵母菌显色培养基法、ITS基因的PCR-RFLP(聚合酶链反应-限制性片段长度多态性)法、ITS测序菌种鉴定法,鉴别真菌各菌株。结果显示:有56例菌株至少通过1种方法检出真菌;玉米吐温-80分离培养假丝酵母菌37株;50例菌株ITS基因测序共鉴定出8个菌种,白假丝酵母菌(C.albicans)29株,近平滑假丝酵母菌(C.parapsilosis)10株,热带假丝酵母菌(C.tropicalis)5株,Candida metapsilosis 1株,Lodderomyces elongisporus 1株,克柔假丝酵母菌(Candida krusei)1株,乙醇假丝酵母菌(C.ethanolica)1株,季也蒙毕赤酵母菌(Pichia guilliermondii)2株;CHROMagar假丝酵母菌显色培养基法鉴定出3种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌;PCR-RFLP法检出5种菌株,分别是白假丝酵母菌、热带假丝酵母菌、近平滑假丝酵母菌、季也蒙毕赤酵母菌、克柔假丝酵母菌,与基因的测序鉴定一致率为91%;糖发酵生化反应法阳性标本占被检出真菌例数的46.4%(26/56)。结果表明:ITS基因的测序法可以准确鉴定真菌各个菌种;PCR-RFLP法能鉴定常见的菌种,但操作繁琐;CHROMagar假丝酵母菌显色培养基法能快速准确鉴别3种常见假丝酵母菌菌种;玉米吐温-80可以准确培养鉴别白假丝酵母菌;糖发酵生化反应法,缺乏足够的敏感度和特异性,难以准确鉴别各个菌种。     

Cell-surface hydrophobicity of Candida species as determined by the contact-angle and hydrocarbon-adherence methods

Cell-surface hydrophobicities of six Candida species were studied by two methods: measurement of the contact angle, and partitioning with aqueous-hydrocarbon (n-octane, n-hexadecane and p-xylene) mixtures. C. tropicalis, C. glabrata and C. krusei adhered better to the hydrocarbons than did C. albicans, C. stellatoidea and C. parapsilosis. Contact angles for the less adherent species were smaller than those for the more adherent species. Thus the two methods gave results that were similar overall and indicated that C. tropicalis, C. glabrata and C. krusei have greater cell-surface hydrophobicities than C. albicans, C. stellatoidea and C. parapsilosis.     

In-vitro killing of Candida species by murine immunoeffectors and its relationship to the experimental pathogenicity

Killing of yeast cells of several species of Candida by murine phagocytic cells was assessed in vitro by a radiolabel release microassay and measurement of colony forming units. The most effective candidacidal phagocytes, i.e. polymorphonuclear and bone marrow cells, were able to kill equally well cells of any species or isolate tested, given sufficient time (4 h) and an appropriate effector: target ratio. However, C. guilliermondii, C. krusei and C. parapsilosis were killed by polymorphonuclear and bone marrow cells much more promptly (1 h) and at a significantly lower effector:target ratio than C. albicans, C. tropicalis and C. viswanathii. Moreover, there were immune effectors such as peritoneal resident macrophages and, mostly, spleen cells which were practically ineffective against C. albicans and C. tropicalis but showed significant activity against C. guilliermondii, C. krusei and C. parapsilosis, even in mice immuno-depressed with cyclophosphamide. Three isolates of C. albicans, differing in the capacity to form germ tubes, also differed in mouse virulence: the germ-tube forming isolate was the most virulent. However, they showed an identical pattern of susceptibility to killing by mouse immunoeffectors, suggesting that virulence is probably not due to the resistance of hyphal cell to phagocytosis.     

In-vitro activity of 5-fluorocytosine against 1,021 Spanish clinical isolates of Candida and other medically important yeasts

The aim of this study was to determine the prevalence of primary resistance to 5-fluorocytosine (5FC) among clinical isolates of yeasts in Spain where this drug is not currently available for therapy. We have tested the in vitro activity of 5FC against 1,021 recent yeast clinical isolates, including 522 Candida albicans, 140 Candida parapsilosis, 68 Candida glabrata, 41 Candida dubliniensis, 50 Candida guilliermondii, 34 Candida tropicalis, 28 Candida krusei, 20 Candida famata, 11 Cryptococcus neoformans, 5 Cryptococcus albidus, 43 Rhodotorula spp., 24 Trichosporon spp., 5 Saccharomyces cerevisiae, 9 Pichia spp., and 21 isolates from other 11 yeast species. The MICs were determined by the ATB Fungus agar microdilution test (bioMerieux, France) and the following interpretive breakpoints were used: susceptible, > 4 microg/ml; intermediate, 8 to 16 microg/ml; resistant, > 32 microg/ml. 5FC was very active against Candida spp. and other medically important yeasts as 852 (83.4%) of the studied isolates were susceptible (MIC < 4 microg/ml). The species most susceptible to 5FC were C. dubliniensis (100%of isolates; MIC90, 0.25 microg/ml), C. famata (100% of isolates; MIC90, 0.25 microg/ml), C. guilliermondii (98%of isolates; MIC90, 0.25 microg/ml), C. glabrata (95.5% of isolates; MIC90, 0.25 microg/ml), and C. neoformans (90.9% of isolates; MIC90, 2 microg/ml). Primary resistance to 5FC was very uncommon, and a MIC > 32 microg/ml, indicator of in vitro resistance, was observed in 106 isolates (10.4%): 77 C. albicans (16.5% of isolates; MIC90, > 128 microg/ml), 9 C. parapsilosis (6.4% of isolates; MIC90, 8 microg/ml), 4 C. albidus (80% of isolates, MIC50, > 128 microg/ml), 3 C. glabrata (4.4% of isolates; MIC90, 0.25 microg/ml), 3 C. tropicalis (8.8% of isolates; MIC90, 4 microg/ml), 2 C. krusei (7.1% of isolates; MIC90, 8 microg/ml), 2 Rhodotorula spp. (4.6% of isolates, MIC90, 1 microg/ml), 8 Trichosporon spp. (33.3% of isolates; MIC90, 64 microg/ml), and 1 C. lipolytica (50% of isolates). Interestingly, most C. albicans (67 out of 77 isolates) resistant to 5FC were serotype B isolates.