申克孢子丝菌基因差异、致病力与孢子丝菌病临床型别的关系

目的 探讨申克孢子丝菌基因差异、致病力与孢子丝菌病不同临床型别的关系。方法 ①收集不同临床型别孢子丝菌病的申克孢子丝菌分离株并提取DNA,进行随机扩增多态DNA(RAPD)扩增。②BALB/c小鼠接种不同临床型别孢子丝菌病的分离株菌悬液,观察实验动物发病及病变情况。③发病小鼠皮肤及内脏组织病理学检查,观察接种不同临床型别孢子丝菌病的分离株菌悬液后小鼠病变内申克孢子丝菌孢子数量及分布。结果 ①不同临床型别孢子丝菌病的申克孢子丝菌分离株聚合酶链反应产物电泳带型差异较明显:播散型分离株可见1800bp、850bp、500bp、180bp,皮肤淋巴管型分离株见1400bp、800bp、700bp、500bp,皮肤固定型分离株见2500bp、1400bp、1000bp、700bp。②注射播散型孢子丝菌病分离株菌悬液的BALB/c小鼠比注射皮肤淋巴管型分离株小鼠发病早、病变部位广且死亡率高;注射皮肤淋巴管型分离株的小鼠较注射固定型孢子丝菌病分离株小鼠皮损出现早、病变范围广且严重。③实验BALB/c小鼠病变皮肤及内脏组织病理学检查显示:注射播散型孢子丝菌病分离株的小鼠病变内孢子数量明显多于注射皮肤淋巴管型分离株小鼠病变内孢子数量,而后者较注射固定型孢子丝菌病分离株的小鼠病变内孢子数量多。结论 不同临床型别孢子丝菌病的申克孢子丝菌的基因差异、致病力与孢子丝菌病不同临床型别的关系密切。     

碘化钾与伊曲康唑联合治疗孢子丝菌病的实验研究

目的观察碘化钾与伊曲康唑单独用药和联合用药对实验性孢子丝菌感染的治疗作用。方法建立小鼠孢子丝菌皮内和系统感染模型,分别设立对照组、碘化钾组、伊曲康唑组及碘化钾和伊曲康唑联合用药组,检测各组真菌培养阳性率、组织中孢子数目及炎症浸润细胞的变化。结果与对照组比较,用药组皮内感染皮损中孢子数目减少,系统感染组织器官真菌培养阳性率降低,联合用药组与单独用药组上述两指标比较均有显著性差异。皮内感染三个用药组都可引起炎症细胞的浸润增加,联合用药组以巨噬细胞的增加尤为明显。结论碘化钾与伊曲康唑联合治疗小鼠孢子丝菌皮内感染和系统感染均优于两种药物单独应用。     

申克孢子丝菌野生株致病性的实验研究

目的 研究申克孢子丝菌野生株的致病性。方法 制备申克孢子丝菌野生株的细胞悬液,注入小鼠体内,于接种后第1周开始至第10周,视发病情况每周分批处死、剖检,观察发病情况。结果 从自然环境中分离的申克孢子丝菌通过不同注射途径进入小鼠体内后均可导致小鼠致病;其产生色素的黑色菌株与不产生色素的白色菌株致病性无差异(P>0.05);腹腔组与尾静脉组比较致病性有差异(P<0.05).结论 申克孢子丝菌的野生株均具有致病性;黑白菌株致病性无显着差异;不同的致病途径致病性不同。     

申克孢子丝菌对小鼠感染后的临床特征与其感染途径及菌本身的致病性有关

孢子丝菌病主要以皮肤及淋巴管型为主,感染肺等内脏甚少见。作者观察了由皮肤(4株)及肺部(3株)孢子丝菌病中分离的菌株对BALB/c小鼠分别进行皮下、静脉及腹腔内攻击以研究其感染机理。     

结合珠蛋白在阿萨希毛孢子菌小鼠皮肤感染中的表达与调控

目的探讨结合珠蛋白(haptoglobin,Hp)在阿萨希毛孢子菌皮肤感染过程中的免疫调节作用。方法 80只BALB/c小鼠分为免疫抑制组和非免疫抑制组,两组小鼠皮下均接种3.2×107cfu/ml的阿萨希毛孢子菌菌悬液,分别于接种后0.5d、1d、3d、5d处死小鼠,取皮损组织,提取总RNA,RT-PCR检测Hp mRNA表达,基因芯片技术研究免疫相关基因差异表达。结果两组小鼠在第0.5d、1d、3d、5d后皮损组织均扩增出Hp mRNA,其中免疫抑制组皮损组织Hp mRNA表达水平明显高于非免疫抑制对照组,差异有显著统计学意义(P<0.01)。免疫抑制组感染1d后皮损组织Hp mRNA表达明显高于0.5d、3d、5d,组间比较均有显著性差异(P<0.01)。筛选出的11条免疫相关差异表达基因中编码结合珠蛋白基因表达上调。结论结合珠蛋白参与了皮肤阿萨希毛孢子菌感染的过程。     

银屑I号对咪喹莫特诱导小鼠银屑病模型核转录因子-κB的调控作用及机制

目的观察银屑I号对咪喹莫特(imiquimod,IMQ)诱导小鼠银屑病模型核转录因子-κB(nuclear factor,NF-κB)炎症通路的影响及其可能的作用机制。方法建立IMQ诱导小鼠银屑病模型,随机分为模型组、银屑I号组、雷公藤多甙组,每组10只,另选正常小鼠10只作为空白对照组,空白对照组及模型组予等量生理盐水灌胃,银屑I号组、雷公藤多甙组分别给予银屑I号、雷公藤多甙灌胃,连续10天,HE染色观察各组小鼠皮损组织形态学变化;免疫组化法检测皮损组织NF-κB p65蛋白水平,RT-PCR法检测NF-κB mRNA表达情况,订制血液蛋白芯片检测血清白介素6(interleukin 6,IL-6)、白介素10(interleukin 10,IL-10)、白介素17(interleukin 17,IL-17)、干扰素γ(interferon gamma,INF-g)含量。结果与空白对照组比较,模型组小鼠出现明显的红斑、鳞屑及皮肤增厚。与模型组比较,银屑I号组、雷公藤多甙组小鼠红斑、鳞屑及皮肤增厚程度明显减轻;银屑I号组、雷公藤多甙组血清IL-6、IL-10、IL-17、INF-γ水平明显降低(P均0.01);银屑I号组、雷公藤多甙组皮损组织NF-κB mRNA、NF-κB p65表达也明显降低(P均0.01)。结论银屑I号能够明显抑制IMQ诱导小鼠模型皮肤增殖,这可能与抑制NF-κB基因表达及减轻炎症反应有关。     

经皮肤损伤感染马尔尼菲青霉菌致病力动物实验研究

目的研究马尔尼菲青霉菌经皮肤损伤途径感染小鼠致病力情况。方法将马尔尼菲青霉菌野生株和人感染株孢子悬液分别注入鼠尾皮内,观察接种后发病情况,于第15,50 d分批处死、解剖。结果马尔尼菲青霉菌导致小鼠皮肤感染发病率为100%,而85%小鼠皮损可自行消退痊愈,15%小鼠出现皮肤播散性感染;组织病理示:皮损处细胞性炎症反应在皮损中显著。从发病时间和早期病变严重程度比较,野生株致病力显著强于人感染株(P<0.05);但是从后期病变严重程度和自行痊愈率比较,野生株和人感染株致病力差异无显著性(P>0.05)。结论马尔尼菲青霉菌可以经皮肤损伤引起小鼠致病,其引起机体剧烈细胞免疫应答反应是致病力重要因素之一;野生株和人感染株感染早期致病力有差异,预后无差异。     

伊曲康唑治疗孢子丝菌病的疗效及抗真菌药敏试验

目的:观察伊曲康唑对孢子丝菌病的临床疗效,检测孢子丝菌对伊曲康唑的体外敏感性.方法:对48例孢子丝菌病患者给予伊曲康唑胶囊口服治疗;另外30例患者给予碘化钾溶液口服治疗,并采用Etest法检测其中10株孢子丝菌对伊曲康唑的最小抑菌浓度(MIC).结果:伊曲康唑对孢子丝菌病的总有效率91.67％,碘化钾溶液的总有效率86.67％,两组总有效率比较无显著性差异,(P＞0.05).采用Etest法检测10株孢子丝菌对伊曲康唑MIC值,范围为0.012-8μg/mL.结论:虽然伊曲康唑对孢子丝菌病的疗效与碘化钾的疗效相当,但副作用小.Etest法可以作为一种孢子丝菌的体外抗真菌药敏试验方法.     

2010—2019年吉林省某三甲医院儿童和成人孢子丝菌病的临床特征比较

【摘要】 目的 比较2010—2019年吉林大学第一医院确诊的儿童和成人孢子丝菌病的临床特征差异。方法 回顾性分析2010年1月1日至2019年12月31日吉林大学第一医院皮肤科3 124例孢子丝菌病患者的临床资料，其中儿童（< 18岁）911例，成人（≥ 18岁）2 213例。所有患者均经皮损真菌培养确诊为孢子丝菌病。采用χ2检验比较儿童与成人孢子丝菌病的临床特征差异。结果 911例儿童患者中，男女比例为1.4∶1，720例（79.0%）发生于寒冷季节（1 - 3月及10 - 12月），790例（86.7%）为皮肤固定型孢子丝菌病，835例（91.7%）皮损发生于面部。2 213例成人患者中，男女比例为1∶2.5，1 450例（65.5%）发生于寒冷季节，1 523例（68.8%）为皮肤固定型孢子丝菌病，1 090例（49.3%）皮损累及四肢，975例（44.1%）累及面部。儿童孢子丝菌病在寒冷季节发病的比例明显高于成人（χ2 = 55.55，P < 0.001），皮肤固定型孢子丝菌病比例亦显著高于成人（χ2 = 107.55，P < 0.001）。两组皮损分布差异有统计学意义（χ2 = 599.91，P < 0.001）。结论 儿童与成人孢子丝菌病的流行病学特征和临床表现存在差异。     

真菌病

林生地霉皮损分离株和血液分离株致病性的实验研究;联苯苄唑乳膏治疗皮肤、黏膜浅部真菌病临床疗效观察;无皮肤损害的幼儿马尔尼菲青霉病2例;利拉萘酯乳膏治疗手足癣和体股癣40例疗效观察;上肢皮肤固定型孢子丝菌病2例     

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。     

申克孢子丝菌酵母相对人急性单核细胞白血病细胞NF-κB信号通路及TNF-α的影响

目的:明确申克孢子丝菌酵母相对人急性单核细胞白血病细胞(THP-1)NF-κB信号通路激活和肿瘤坏死因子α(TNF-α)分泌的影响。方法:实时荧光定量PCR分析申克孢子丝菌酵母相刺激THP-1细胞TNF-αmRNA的表达,酶联免疫吸附法检测TNF-α分泌量,免疫印迹法分析申克孢子丝菌酵母相体外作用于THP-1细胞后IκBα和磷酸化IκBα的水平,免疫荧光法观察NF-κB-p65核转位。同时设置地塞米松(NF-κB抑制剂)抑制组,并检测100 n M地塞米松预处理THP-1细胞后TNF-αmRNA水平的变化。结果:申克孢子丝菌酵母相刺激THP-1细胞后6 hTNF-αmRNA水平显著高于空白对照组(P0.001)。申克孢子丝菌酵母相刺激THP-1细胞后24 h,TNF-α蛋白水平为(4610.419±121.501)pg/mL,显著高于空白对照组(186.964±98.073)pg/m L,差异具有统计学意义(P0.001)。申克孢子丝菌酵母相作用THP-1细胞后30~60 min,磷酸化IκBα蛋白水平显著升高,为时间依赖性。申克孢子丝菌酵母相组细胞核内NF-κB-p65较空白对照组荧光强度增强。100 nM地塞米松预处理各组THP-1细胞后,TNF-αmRNA水平较前明显降低。结论:人THP-1细胞体外与申克孢子丝菌酵母相作用后激活NF-κB信号通路并上调TNF-α分泌。     

Zoonotic sporotrichosis of lymphocutaneous type in a man acquired from a domesticated feline source: report of a first case in southern Karnataka, India

Background  Sporotrichosis is commonly encountered due to traumatic implantation of thorns or decayed vegetation with the dimorphic fungi, Sporothrix schenckii . Zoonotic spread of Sporotrichosis is rare and we describe here the first case of feline transmission of lymphocutaneous sporotrichiosis encountered in India.
Methods  An excision biopsy of nodulo-ulcerative lesion from the patients right elbow and forearm were collected for histopathology and portion of the specimen processed for mycological work up. Animal pathogenicity test performed in Swiss albino mice with intraperitoneal & foot pad inoculation. In addition an investigation of the ulcerative skin lesion from the domesticated cat was carried out.
Results  Histopathology examination of tissue sample from the patient and feline lesion revealed granulomatous reaction and a few slender elongated yeast cells consistent with Sporotrichosis. The diagnosis was confirmed by culturing Sporothrix schenkii and demonstration of thermal dimorphism. Pathogenicity testing in mice lead to orchitis in 12–15 days and the organism was re-isolated in pure culture. The patient was treated with oral saturated potassium iodide solution with complete resolution of the lesions.
Conclusion  Close contact with infected domesticated feline can be a potential source of transmission for Sporotrichosis as evidenced in this report.     

Potassium iodide remains the most effective therapy for cutaneous sporotrichosis

Sporotrichosis is a subcutaneous fungal infection caused by the dimorphic fungus Sporothrix schenckii. Itraconazole has largely replaced older therapies, but we present a case of lymphocutaneous sporotrichosis that failed to respond to an adequate course of itraconazole yet responded dramatically to treatment with saturated solution of potassium iodide (SSKI).     

Sporotrichosis in Nepal

The first case of sporotrichosis from Nepal is reported in a 25-year-old man from a village about 60 km east of Kathmandu. He never travelled outside of Nepal before and had acquired the lymphocutaneous form of the disease after an accidental injury to the right foot while cutting wood. The diagnosis of the case was made by culturing Sporothrix schenckii from the lesions, proving the dimorphic character of the fungus in vitro, its pathogenicity in mice, and its serology. Oral potassium iodide therapy resulted in complete cure.     

Sporotrichosis in a husband and wife

Familial occurrence of sporotrichosis is rare. We report two patients, a husband and wife, with sporotrichosis. A 67-year-old man and a 63-year-old woman, who were farmers, had erythematous plaques and nodules on their faces and his forearm. They had noticed their cutaneous lesions during nearly the same period without any traumatic history. Fragments of biopsy specimens from both of them were submitted for mycological cultures and yielded pure cultures of Sporothrix schenckii (S. schenckii). Restriction fragment length polymorphisms in the mitochondrial DNA of the S. schenckii isolated from the biopsy specimens of their lesions were investigated. The isolates were identified as type 5, which is comparatively abundant in the Kanto area in Japan. The husband was treated with potassium iodide and itraconazole. His wife was treated with itraconazole alone. We failed to isolate a causative fungus from the soil. Our case is the second case in the literature of sporotrichosis in a husband and wife during the same period.     

Sporotrichosis

Sporotrichosis is the most common subcutaneous mycosis. It is usually acquired by traumatic inoculation, and it is caused by one of the species of the Sporothrix schenckii complex. More than 6 species, such as S schenckii sensu stricto, Sporothrix brasiliensis, Sporothrix globosa, Sporothrix mexicana, and Sporothrix albicans, have been identified by molecular techniques. The most common presentation is cutaneous disease, which is classified into fixed and lymphocutaneous forms. Osteoarticular, pulmonary, mucosal, disseminated, and systemic infections are less common and usually occur in immunosuppressed individuals. The diagnosis is suggested by biopsy specimen and confirmed by tissue culture. Itraconazole is considered the treatment of choice, although in some undeveloped countries potassium iodide is still used, owing to its safety and low cost. For systemic or disseminated cases, amphotericin B is the treatment of choice.     

“ITS/CAL"靶位分子鉴定结合表型分析确诊狭义申克孢子丝菌致淋巴管型孢子丝菌病一例

患者女,54岁,农民.左上肢近手腕部外伤后出现豌豆大小的单一红色结节15d,后肿大、破溃,并在30 d内发展为多个成串状分布的结节.皮肤科检查:患者左上肢可见多个呈线性排列的紫红色结节,质硬;部分结节破溃,伴少许脓性分泌物.组织病理提示,感染灶呈混合炎性细胞浸润为主的化脓性肉芽肿炎症;过碘酸雪夫染色阴性,未见真菌孢子、菌丝及星状体.活检组织真菌培养阳性.根据培养物形态学分析和内部转录间隔区(ITS)及钙调蛋白(CAL)编码区靶位的分子生物学鉴定结果,确诊该病例为狭义申克孢子丝菌致淋巴管型孢子丝菌病.给予患者10％碘化钾溶液10 ml/次每日3次口服;治疗2个月后,患者自觉症状明显改善,后失访.本例报道提示联合应用表型鉴定和“ITS/CAL”靶位的基因分析能够准确将孢子丝菌复合体鉴定到种水平.     

碘化钾治疗孢子丝菌病的实验研究

对不同免疫状态小鼠造成孢子丝菌实验性感染，观察碘化钾的治疗作用和病损发展过程中病原体与炎症浸润细胞的变化及两者间的关系。结果进一步证实碘化钾对孢子丝菌病的治疗作用与宿主的免疫力有关，并发现碘化钾具有促进巨噬细胞在感染部位的浸润和增强小鼠对孢子丝菌的抵抗力，还表明碘化钾对小鼠内脏感染无效，而对血源播散型皮肤感染的作用因小鼠的免疫状态而异。