须癣毛癣菌肉芽肿株与体癣株形态学、致病性及比较蛋白质组学的差异性研究

目的 筛选须癣毛癣菌肉芽肿株和体癣株的差异表达蛋白,探讨须癣毛癣菌致深部和浅部感染的发病机制.方法 选取4株肉芽肿株和4株体癣株须癣毛癣菌,行27℃和37℃平皿及钢圈法小培养,观察形态变化.取8只豚鼠,糖皮质激素干预改变豚鼠的免疫状态,每只豚鼠对称部位分别采用表面和皮下两种接种方式接种同一种菌,制备体癣模型和肉芽肿模型,接种后2周左右直接镜检及培养、组织病理检查,验证感染效果.筛选肉芽肿株强毒株和体癣株弱毒株各1株,行双向电泳,质谱检测和NCBI数据库分析.结果 须癣毛癣菌37℃培养,4株肉芽肿株较4株体癣株生长良好;27℃培养,两者形态差别不明显.豚鼠感染实验,所有菌株均构建了体癣模型,4株肉芽肿株较4株体癣株炎症反应明显.3株肉芽肿株形成皮下结节,直接镜检及培养、组织病理检查均显示感染成功.比较蛋白质组学,肉芽肿株和体癣株分别检测到蛋白点(463±20)个与(398±17)个,肉芽肿株有62个蛋白上调表达,体癣株有21个蛋白上调表达,肉芽肿株中有致病意义的蛋白包括烯醇化酶、热休克蛋白家族、丝-苏氨酸蛋白酶、细胞信号转导相关蛋白、能量相关蛋白、细胞骨架蛋白及一些假想蛋白.结论 须癣毛癣菌肉芽肿株较体癣株耐热,易感染宿主;两者蛋白质水平有明显差异.     

须癣毛癣菌和犬小孢子菌共同致面癣1例

报道罕见由须癣毛癣菌和犬小孢子菌共同感染引起的面癣1例。患者女性,70岁,反复左面部红斑伴瘙痒1年。取皮损鳞屑直接真菌镜检可见菌丝,培养分离鉴定为须癣毛癣菌和犬小孢子菌。给予口服特比萘芬及外用抗真菌软膏3周后治愈。随访3月无复发。     

167份指趾甲真菌感染直接镜检和培养结果分析

目的了解本地区指趾甲真菌感染的情况。方法对皮肤性病科门诊采集的167份指趾甲真菌感染标本进行直接镜检和真菌培养。结果真菌直接镜检指甲(68/94)、趾甲(53/73)阳性;培养阳性指甲(73/94)、趾甲(58/73),其中红色毛癣菌81(61.83%);须癣毛癣菌21(16.03%);絮状表皮癣菌4(3.05%);犬小孢子菌2(1.53%);假丝酵母菌18(13.74%);铁锈色小孢子菌2(1.53%);玫瑰色毛癣菌2(1.53%);短帚霉1(0.76%)。结论本地区指趾甲真菌感染以红色毛癣菌为优势菌占第一位,第二位须癣毛癣菌,第三位假丝酵母菌。以皮肤癣菌感染为主,酵母样菌感染较少。     

196例儿童头癣病原菌分析

目的了解湖北仙桃地区近16年来儿童头癣的发病及病原菌分布状况。方法对拟诊为头癣的患儿、取病发及鳞屑作真菌直接镜检及真菌培养。结果共分离出病原菌13种,致病菌株196株,主要病原菌为堇色毛癣菌、石膏样毛癣菌、红色毛癣菌、羊毛状小孢子菌、断发毛癣菌等。病原菌主要分布于5-12岁。     

开远地区10年浅部真菌及病原菌种类分析

目的为掌握开远地区的浅部真菌病情况及病原菌菌种分布特点。方法对临床拟诊浅部真菌病的患者3358人次，取标本行10％KOH涂片镜检、分离培养及菌种鉴定。分析浅部真菌病的发病情况。结果直接涂片查见菌丝和／或孢子的2249份，阳性率67％。培养出真菌1356株，阳性率40．4％。其中红色毛癣菌754株、须癣毛癣菌154株、念珠菌210株、犬小孢子菌62株、铁锈色小孢子菌104株、石膏样小孢子菌18株、疣状毛癣菌14株、玫瑰色毛癣菌2株、紫色毛癣菌2株、断发毛癣菌4株。结论开远地区的浅部真菌病病原菌以红色毛癣菌为优势菌，其次为念珠菌，再次为须癣毛癣菌。     

浅部真菌病668例病原菌种类分析

目的分析我地区浅部真菌病致病菌及菌种构成情况。方法通过对近2年我院浅部真菌病患者临床送检标本进行直接镜检、培养及真菌鉴定,鉴定到种。结果 1 600份临床送检标本,668份直接涂片镜检阳性,阳性率41.75%,其中甲真菌病214例、体癣176例、足癣166例、手癣78例;将阳性标本进行真菌培养,共培养分离207株致病菌,培养阳性率30.98%,其中红色毛癣菌居首位,须癣毛癣菌第2位,犬小孢子菌居第3位。结论河南地区浅部真菌病主要为甲真菌病、体癣、足癣,致病菌种以红色毛癣菌为主。     

疣状毛癣菌致母女共患体癣分析

母亲上肢出现皮损1年，女儿面部皮损4个月，皮损表现为大小不一的环形红斑，表面覆有鳞屑及少许痂皮，境界清楚，边缘稍隆起。患者家中养牛。两例皮损真菌镜检均可见菌丝，真菌培养均阳性，分离菌株经表型及分子鉴定，两株菌均为疣状毛癣菌，体外药敏试验显示对伊曲康唑及特比萘芬均敏感。 2例患者经口服和外用抗真菌药物联合治疗获痊愈。     

氮掺杂碳纳米酶联合近红外光抗真菌体外试验研究

目的：评价氮掺杂碳纳米酶（N-Carbon nanozyme）联合近红外光对白念珠菌和红色毛癣菌体外抗菌的效果。方法：将白念珠菌和红色毛癣菌标准菌株常规接种培养，分别分为四组：(1)空白对照组：真菌不作任何处理；(2)光照组：真菌予以2.5 W/cm2近红外光照射8 min；（3）材料组：真菌予以250 μg/mL的N-Carbon nanozyme处理30 min；（4）实验组：真菌予以250 μg/mL的N-Carbon nanozyme及2.5 W/cm2近红外光照射8 min。SYTO 9/PI染色判断白念珠菌和红色毛癣菌的菌体活性；扫描电镜观察和平板计数法判定白念珠菌抗菌形态和菌落形成单位（CFU）的数量。结果：实验组中SYTO9/PI染色示红色荧光染色信号显著增加；与空白对照组比较，光照组、材料组间菌落数均未见明显差异，而实验组可见菌落明显减少。生物扫描电镜显示白念珠菌的细胞形态发生明显的变化。结论：氮掺杂碳纳米酶联合近红外光对白念珠菌及红色毛癣菌均有明显的抑制作用。     

北京地区儿童头癣的病原菌分析

为了解北京及其周边地区的头癣流行状况及病原菌分布，对508例头癣患者病发均做了真菌直接镜检和真菌培养，结果直接镜检阳性423例（83．3％），培养阳性357例（70．3％），分离出犬小孢子菌263例（73．7％），须癣毛癣菌72株（20．1％），红色毛癣菌8株（2．2％），石膏样小孢子菌7株（2．0％），紫色毛癣菌6株（1．7％），铁锈色小孢子菌1株（0．3％）。结果示头癣是儿童常见的真菌感染性皮肤病，在北京地区主要致病菌为犬小孢子菌和须癣毛癣菌，未发现黄癣菌。     

狐狸饲养者母女共患须癣毛癣菌所致体癣的临床与实验研究

目的探讨狐狸饲养者母女先后出现同形皮疹的病因及病原菌的实验研究。方法刮取两名患者皮屑直接镜检;皮屑与其饲养的狐狸被毛同时做真菌培养,并对病原菌行PCR扩增测序。结果三株菌在沙堡氏葡萄糖琼脂培养基﹙SDA﹚上生长为白色颗粒状菌落;尿素酶试验均阳性;扫描电镜可见分隔菌丝及大分生孢子;菌落PCR扩增测序结果与须癣毛癣菌100%同源。结论两患者体癣病原菌与狐狸被毛所携带的须癣毛癣菌为同一来源。     

汉防己甲素对联苯苄唑抗须癣毛癣菌活性增效作用的体内实验研究

目的研究汉防己甲素(tetrandrine,TET)对联苯苄唑(bifonazole,BFZ)治疗须癣毛癣菌所致豚鼠皮肤癣菌病是否有增效作用。方法以须癣毛癣菌构建豚鼠皮肤癣菌病模型,将造模成功的20只豚鼠按随机数字表法等分为4组,进行双盲实验,分别涂用复方BFZ(含1%BFZ和1%TET)、1%BFZ、1%TET和基质,1次/d,共14天,进行皮损评分、病原学和病理学检查,分析4组药物间疗效差异。结果用药期间,4组皮损评分差异有统计学意义(F=93.49,P=0.000);从用药第3天起,复方BFZ组和1%BFZ组皮损评分逐日下降,前者皮损评分持续低于后者,差异有统计学意义(P=0.000);1%TET组和基质组皮损评分差异无统计学意义(P=1.000)。复方BFZ组皮损真菌学检查比1%BFZ组转阴早2天,炎症反应轻于其他3组。结论TET对BFZ治疗须癣毛癣菌所致豚鼠皮肤癣菌病有增效作用。     

Experimental systemic dermatophytosis

Albino guinea pigs with abraded skin were inoculated cutaneously with 27 strains of dermatophytes from animals and humans. The same strains were inoculated i.v. in guinea pigs with intact skin. Zoophilic dermatophytes and human isolates of Trichophyton mentagrophytes var. granulare produced ringworm after cutaneous application. After i.v. inoculation, the fungus was reisolated from skin samples from a considerable number of animals with and without clinical ringworm lesions, and also from lungs, liver, and kidneys. In the lungs, hyphal aggregates were noted. Some strains produced generalized dermatophytosis affecting all parts of the skin and internal organs. T. mentagrophytes B32663, selected for further study, was inoculated i.v. in the guinea pig, rabbit, rat, mouse, and chicken. Ringworm lesions occurred in the guinea pig and rabbit; in the mouse, rat, and chicken other organs were involved. Administration by other routes did not produce a generalized infection. The infection was not self-limited and the deep-seated lesions may be responsible for the recurrence of infection.     

皮肤癣菌感染动物模型的建立及抗真菌治疗的评价

目的 建立皮肤癣菌感染动物模型并评价抗真菌药物对该模型的体内疗效。方法 选择38只健康豚鼠,随机分为须癣毛癣菌感染组和犬小孢子菌感染组,用磨砂法建立皮肤癣菌感染的动物模型,每组各有1只为阴性对照。每组18只感染动物再随机分为伊曲康唑治疗组、特比萘芬治疗组、未治疗组3个组。在治疗后的第8、11、14天分别对动物模型进行皮肤病变评分和真菌学检查。结果 未进行治疗的感染动物均出现明显的皮损,且真菌学检查为阳性。须癣毛癣菌感染的伊曲康唑治疗组动物皮损评分和真菌学治愈率在第8、11、14天分别为9、1、0,66.7%、83.3%、83.3%;特比萘芬治疗组分别为8、5、1,83.3%、83.3%、83.3%;未治疗组分别为48、52、40,0%、0%、0%。犬小孢子菌感染的伊曲康唑治疗组动物皮损评分和真菌学治愈率在第8、11、14天分别为3、0、0,83.3%、83.3%、83.3%,特比萘芬治疗组分别为9、2、0,83.3%、83.3%、83.3%,未治疗组分别为46、47、39,0%、0%、0%。两组与未治疗组比较,差异均有统计学意义（P < 0.01）,但伊曲康唑治疗组与特比萘芬治疗组比较差异无统计学意义（P > 0.05）。结论 伊曲康唑治疗须癣毛癣菌和犬小孢子菌感染的动物模型有良好的疗效,特比萘芬有相似的结果。     

Trichophyton mentagrophytes spores differ from mycelia in their ability to induce pustules and activate complement

To determine whether the morphology of dermatophytes plays a role in the clinical manifestation of dermatophytosis, we isolated spores and mycelia from colonies of killed Trichophyton mentagrophytes and tested their ability to induce gross and microscopic cutaneous changes when applied to guinea pig skin. The skin of normal guinea pigs failed to develop any cutaneous changes after inoculation with either dead spores or mycelia. In contrast, guinea pigs that had recovered from a T. mentagrophytes infection reacted to topically applied spores by producing pustules but failed to develop pustules either grossly or microscopically when inoculated with mycelia. Because differences in the activation of C' and the subsequent generation of the polymorphonuclear leukocyte (PMN) chemoattractant, C5a, might account for our in vivo findings, we measured the amount of C5a activated in vitro when serum was incubated with equivalent amounts of spores or mycelia. Spores consistently activated more C' to C5a than mycelia, although each could activate serum C'. The results support previous studies that show a direct correlation between the clinical manifestations of a dermatophytic infection and the development of immunity against the dermatophyte. Furthermore, the data suggest that a dermatophyte's predominant morphology during an infection may play a critical role in producing pustules by activating C' and chemoattracting PMNs into the fungus-laden stratum corneum.     

The Effect of Aqueous Garlic Extract on the Growth of Dermatophytes

ABSTRACT: The effect of aqueous garlic extract on the growth of different medically important dermatophytes was studied. The extract proved to have an inhibitory effect on the growth of Microsporum gypseum, Trichophyton verrucosum, Trichophyton violaceum, Trichophyton rubrum, Trichophyton schoenleini, Trichophyton mentagrophytes and Epidermophyton floccosum. The inhibitory concentration ranged from 130-200 mg/1. Experimental infection was induced in guinea pigs and rabbits. Treatment of the produced lesions included topical application or injection of the extract. The topical treatment was very effective, producing complete healing in 14 to 17 days following twice daily application of the extract for one week.     

自噬对水痘-带状疱疹病毒感染豚鼠背根神经节模型中病毒结构及功能蛋白表达的影响

【摘要】 目的 探讨自噬诱导剂雷帕霉素及抑制剂3-甲基腺嘌呤对水痘-带状疱疹病毒（VZV）感染豚鼠背根神经节模型中病毒结构及功能蛋白生物合成的影响及机制。方法 在人胚肺成纤维细胞（HELF）中培养扩增VZV,同时分离提取豚鼠外周血单个核细胞（PBMC）,VZV-HELF与PBMC共培养18 ~ 20 h,离心收集VZV-PBMC。32只豚鼠随机平均分为4组,每组8只：空白对照组于内眦静脉丛注射自体PBMC;自噬抑制组、自噬诱导组、VZV感染组分别先腹腔注射3 mg/kg 3-甲基腺嘌呤溶液、0.5 mg/kg雷帕霉素溶液、相同体积的0.9% NaCl溶液,2 h后均于内眦静脉丛注射VZV-PBMC 50 μl。14 d后,处死各组豚鼠,取背根神经节组织,透射电镜观察病毒颗粒形态及自噬囊泡形态及数目,Western印迹检测VZV核衣壳蛋白（NCP）和立即早期蛋白62（IE62）及自噬相关蛋白Beclin-1和p62的表达,免疫组化检测VZV感染的背根神经节中抗VZV抗体的表达。统计分析采用两独立样本t检验、单因素方差分析、LSD-t检验或Kruskal-Wallis H检验。结果 透射电镜下,VZV感染组背根神经节中可见核衣壳病毒粒子及散在的自噬体结构。空白对照组、VZV感染组、自噬诱导组及自噬抑制组自噬囊泡数量［M（Q1,Q3）］分别为0、5（4,6）、7（5,9）、0个,差异有统计学意义（H = 135.60,P＜0.01）,VZV感染组显著高于空白对照组及自噬抑制组（均P＜0.05）,但与自噬诱导组差异无统计学意义（P＞0.05）,而自噬诱导组显著高于自噬抑制组（P＜0.05）。Western印迹显示,VZV感染组IE62蛋白表达水平（1.49 ± 0.06）显著高于空白对照组（0.50 ± 0.09）,t = 9.17,P＜0.05;自噬抑制组抗VZV抗体表达显著低于自噬诱导组和VZV感染组（t值分别为9.24、7.78,均P < 0.01）,而自噬诱导组与VZV感染组抗VZV抗体表达差异无统计学意义（P > 0.05）。结论 VZV感染豚鼠背根神经节后发生了自噬;通过抑制自噬,豚鼠背根神经节中部分VZV结构及功能蛋白的表达水平下降。     

甘草酸软膏治疗豚鼠慢性湿疹模型的实验研究

目的 观察甘草酸软膏抗炎、止痒和对豚鼠慢性湿疹的治疗作用。方法 雄性昆明种小鼠、雌雄昆明种小鼠、白色豚鼠各60只,分别随机分为模型对照组（10只）、基质对照组（10只）、醋酸地塞米松乳膏组（10只）、甘草酸软膏高（10只）、中（10只）、低（10只）剂量组,各组分别建立二甲苯致小鼠耳廓肿胀模型,右旋糖酐40致小鼠搔抓模型,以及2,4-硝基氯苯反复刺激的慢性湿疹豚鼠模型,分别观察甘草酸软膏对小鼠耳廓肿胀和搔抓反应以及慢性湿疹豚鼠耳廓肿胀的影响。结果 甘草酸软膏高、中剂量可以抑制二甲苯致小鼠耳廓肿胀（P < 0.05或0.01）;甘草酸软膏高剂量可以延长右旋糖酐致小鼠搔抓的潜伏期,甘草酸软膏高、中、低剂量可减少搔抓次数（P < 0.05或0.01）,减轻慢性湿疹豚鼠模型耳廓的肿胀（P < 0.05或0.01）,并对耳廓皮肤的病理改变有一定的改善作用。结论 甘草酸软膏对慢性湿疹豚鼠模型有一定的治疗作用。     

Response of guinea pigs to intravaginal inoculation with guinea pig cytomegalovirus

Female guinea pigs were inoculated intravaginally with guinea pig cytomegalovirus (GPCMV) propagated either in guinea pig embryo fibroblast cultures (GPEF) or salivary glands. The incidence of infection was higher with GPEF virus. Rare instances of isolation of GPCMV from cervical swabs 9-48 hr after inoculation was attributed to survival of inoculum in the genital tract. Neither immunofluorescence microscopy nor histopathologic examination showed evidence for active infection of genital tract tissue examined up to day 5 after inoculation. At necropsy on days 30-49, GPCMV was isolated from salivary glands and occasionally from pancreas and lymph nodes. Seroconversion following intravaginal inoculation was demonstrated by an enzyme-linked immunosorbent assay (ELISA) test, and titers were comparable to those after intraperitoneal or subcutaneous inoculation. However, titers of neutralizing antibody, determined by plaque-reduction assay, were significantly lower in the group inoculated intravaginally. These findings are relevant to consideration of cytomegalovirus as a sexually transmitted agent in humans.     

成纤维细胞生长因子10单克隆抗体对豚鼠银屑病样模型的作用研究

目的 探讨成纤维细胞生长因子（FGF10）单克隆抗体局部外用对银屑病豚鼠模型的治疗作用。方法 应用5%盐酸普萘洛尔搽剂外涂豚鼠耳背部皮肤,诱导银屑病样动物模型。分别设置空白组、模型组、丁酸氢化可的松治疗组、FGF10抗体高（0.188 g/L）、中（0.094 g/L）、低（0.063 g/L）剂量治疗组。治疗2周后观察银屑病样模型的病理变化。采用SPSS 13.0统计软件进行统计学分析,Baker评分比较应用秩和检验,单一核细胞计数及表皮厚度比较应用方差分析（ANOVA）,多个样本均数的两两比较应用最少显著差法（LSD）。 结果 各治疗组上述各项检测指标均低于模型组（均P < 0.05）。FGF10抗体高剂量组炎细胞计数与空白组差异无统计学意义（t = 0.77,P > 0.05）,其余各治疗组炎细胞计数高于空白组及FGF10抗体高剂量组（均P < 0.05）。FGF10抗体各治疗组表皮厚度均高于丁酸氢化可的松治疗组,差异均有统计学意义（均P < 0.05）。FGF10抗体各治疗组组间表皮厚度比较,差异无统计学意义（均P > 0.05）。结论 FGF10单克隆抗体对银屑病样豚鼠模型中异常病理改变有显著调节作用,能影响角质形成细胞增殖分裂,还能显著抑制银屑病模型中炎症反应。     

The dynamics of dermatomycetes in the Bashkir ASSR in 1979-1987

Analysis of changes in dermatomycetes incidence in Bashkiria in 1979-1987 has shown the predominance of zoophilic fungi in the latest 9 years, while the share of anthroponotic fungi has made up but 8.6%. The major agent of dermatophytosis in the republic was Microsporum canis. The incidence of Trichophyton verrucosum and Trichophyton mentagrophytes v. gypzeune has grown of late, this necessitating the detection of new epidemiologic factors, contributing to dissemination of these mycoses and the development of appropriate prophylactic measures.