面部原发性皮肤隐球菌病1例

报道1例自身免疫功能正常的原发性皮肤隐球菌病。患者因"面部红斑进行性加重半年,破溃伴结痂2个月余"来我科就诊,皮肤科检查可见左侧面颊部3 cm×5 cm大小红斑,边界清楚,中央可见数片浅溃疡,覆盖灰褐色痂皮。病理检查示真皮浅层及深层可见大量炎性细胞浸润,PAS染色见大量紫红色孢子聚集在多核巨细胞内外;阿申蓝染色阳性;粘蛋白卡红染色阳性;免疫组化染色(抗体为兔抗隐球菌抗体)可见棕色圆形孢子。根据其临床症状、病理检查、特殊染色及免疫组化染色,确诊为原发性皮肤隐球菌病。经伊曲康唑治疗痊愈。     

肝移植后播散性隐球菌病1例及其实验研究

目的播散性隐球菌病临床及实验研究。方法患者女,47岁,肝移植术后2 d,面部、肩部、四肢皮肤出现多发溃疡,伴昏迷。通过脑计算机断层扫描、皮损组织病理检查、PAS染色、皮损组织真菌培养及激光俘获显微切割结合PCR扩增序列分析确诊,并对获得菌株进行尿素酶试验、API试验、PCR扩增测序等实验研究。结果皮损组织病理可见大量圆形和椭圆形菌体,PAS染色阳性。血液和脑脊液真菌镜检均为阴性。皮损组织真菌培养可见酵母样菌落生长,菌株尿素酶试验阳性,API试验鉴定为新生隐球菌。ITS区序列分析鉴定为新生隐球菌grub ii变种。激光俘获显微切割结合PCR扩增,序列分析与培养获得的菌株直接PCR扩增后序列分析结果一致。脑脊液特异性隐球菌抗原（＋＋）,血液特异性隐球菌抗原（＋＋＋＋）。脑CT显示为多发结节灶。依据临床及实验室检查确诊为播散性隐球菌病,致病菌为新生隐球菌grubii变种。结论通过对该病例的深入研究,为临床明确诊断播散性隐球菌病奠定基础,确立了显微切割技术在皮肤真菌感染中的应用价值。     

2000～2008年原发性皮肤隐球菌病回顾性分析

目的了解2000～2008年间原发性皮肤隐球菌病(PCC)的流行病学及临床特征。方法检索并回顾性分析2000～2008年文献报道的原发性皮肤隐球菌病病例资料,对比分析2000～2008年与1985～2000年报道的PCC病例及免疫正常与免疫受损PCC病例的特征是否存在差异。结果共检索出2000～2008年间符合PCC诊断标准的病例28例。近8a来PCC仍好发于年龄较大者,但男性更多见;免疫正常或受损宿主均可发生PCC;HIV感染患者发生PCC几率更小;外伤可能为PCC重要的诱发因素;皮疹部位仍以四肢等暴露部位为主;皮疹性质由以前较特异的化脓性指头炎或蜂窝织炎为主变迁到以溃疡、结节、红斑、肿块等非特异性皮疹为主;治疗仍以抗真菌药物口服或与手术联合为主,氟康唑仍为一线药物;预后一般较好。结论近年来PCC表现出一些新的特点。宿主的免疫状态可能不影响其发生PCC的机率或影响较小,而仅影响隐球菌感染后的转归。     

深在型皮肤念珠菌病1例报告

深在型皮肤念珠菌病一般包括念珠菌肉芽肿（Candidal granuloma）和慢性皮肤黏膜念珠菌病两种。前者临床表现为增生、结节、溃疡或肉芽肿形成。国内1964年张永圣[1]等报道1例后,陆续有病例报道。该病有两种类型：Houser-Rothman型和Busse-Buschke型。Houser-Rothman型也被认为属于慢性皮肤黏膜念珠菌病的一种较严重表现[2]。本文报告1例面部的深在型皮肤念珠菌病。     

念珠菌应激传导通路研究进展

念珠菌的无性阶段是属于隐球菌酵母目,隐球酵母科的一种临床上重要的致病真菌,其有性阶段属于子囊菌亚门。在临床上,念珠菌致病病谱广,它可引起皮肤、黏膜及内脏系统的感染。近年来,由于肿瘤、AIDS患者的增多,念珠菌感染的发病率均呈上升趋势。在美国的一项调查中,发现念珠菌引起的院内感染居第四位,其引起的死亡率高。由于上述特点,所以念珠菌越来越引起人们的重视。     

第一届隐球菌与隐球菌病学术研讨会会议纪要

隐球菌病是由隐球菌属中桌些种或变种引起的一种深部真菌病,主要侵犯中枢神经系统,预后严重,死亡率高,也可侵犯肺、皮肤、骨骼等其他器官．．近年来,隐球菌感染发生率呈明显上升的趋势,已成为一种严重危害人类健康的疾病,对隐球菌与隐球菌病的研究已受到全球真菌学者的高度重视,目前已召开了6届“隐球菌与隐球菌病”国际专题会议商讨对策类似的专题会议在国内目前还尚未开展.     

ATB FUNGUS2法在念珠菌属和新生隐球菌抗真菌药敏试验中应用的评价

目的评价ATBFUNGUS2半固体培养基法在测定念珠菌属和新生隐球菌对4种常用抗真菌药物敏感性中的应用价值。方法利用CLSIM27．A2微量液基稀释法和ATBFUNGUS2法同时测定131株念珠菌和20株新生隐球菌对两性霉素B（AmB）、氟康唑（FLC）、氟胞嘧啶（5-Fc）和伊曲康唑（ITC）的敏感性。结果①两种方法对于AmB、5-FC、FLC和ITC的一致性分别为98％、89．4％、78．8％和78．1％;②所有受试菌株中两种方法的一致性为80％,但ATBFUNGUS2法将2／5株M27-A2法检查为FLC耐药的白念珠菌判断为敏感或剂量依赖,将8／10株M27-A2法检查为FLC剂量依赖的白念珠菌判断为敏感或耐药。③ATBFUNGUS2法中AmB的MIC值判读范围偏高,以致于实际工作中不能读出准确的值。结论ATBFUNGUS2半固体培养基法在测定念珠菌属和新生隐球菌对4种常用抗真菌药物的敏感性时不失为简单、快速而且重复性好的方法。     

1557株酵母菌的鉴定及其药敏试验分析

目的 研究临床分离的酵母菌种类及其对药物的敏感性。方法 采用API 20C、显色培养基对2004-2005年北京协和医院细菌室分离到的1557株念珠菌进行菌株鉴定,并对2005年收集的酵母菌采用ATB FUNGUS 2对氟康唑,伊曲康唑,两性霉素B,5-氟胞嘧啶进行药物敏感性试验;并以whonet 5．3软件进行结果分析。结果 在1557株酵母菌中自念珠菌为57．5％,热带念珠菌为17．0％,光滑念珠菌为14．8％,克柔念珠菌为2．8％,近平滑念珠菌为2．3％,葡萄牙念珠菌为0．9％,新生隐球菌为0．7％,季也蒙念珠菌0．4％,其他菌株共为3．6％。药敏结果显示：近平滑念珠菌、白念珠菌、热带念珠菌、光滑念珠菌、克柔念珠菌和其他念珠菌对氟康唑的敏感率分别为100％、97．6％、94．5％、81．1％、9．1％和75．0％;对5-氟胞嘧啶的敏感率分别为100％、98．0％、98．0％、99．0％、9．5％和88．0％。伊曲康唑对克柔念珠菌和未鉴定到种的念珠菌的MIC90为1μg／ml,光滑念珠菌为4μg／ml,热带念珠菌为0．5μg／ml,白念珠菌和近平滑念珠菌均为0．125μg／ml;两性霉素B对所有念珠菌的MIC50和MIC90均为0．5μg／ml。结论 白念珠菌比例最高,其次为热带念珠菌及光滑念珠菌。5-氟胞嘧啶和氟康唑对除克柔念珠菌之外的所有念珠菌敏感度都很高,两性霉素B对所有念珠菌的MIC90都很低,伊曲康唑对除克柔念珠菌和光滑念珠菌之外的其他菌株的MIC90较低。     

肝肾移植术后患者隐球菌感染的临床特征分析

目的研究肝、肾移植术后患者隐球菌感染的临床特征。方法选取2010年1月到2015年7月来浙江大学医学院附属第一医院就诊的肝、肾移植术后并确诊为隐球菌感染的患者,对其临床特征进行回顾性分析。结果研究周期内有23例患者符合入组要求,其中肾移植术后21例、肝移植术后2例。对23例患者进行分析,发现单纯隐球菌肺炎6例(占26.0%)、单纯隐球菌性脑膜炎6例(占26.0%)、隐球菌性脑膜炎合并隐球菌肺炎8例(占34.7%)、隐球菌败血症2例(占8.6%),皮肤隐球菌感染1例(占4.3%)。所有隐球菌肺炎均经肺穿刺病理确诊,临床表现以发热,咳嗽咳痰,气急症状居多。胸部CT表现为结节、空洞、肿块、渗出等。所有隐球菌脑膜炎患者中9例经脑脊液培养出新生隐球菌、7例脑脊液墨汁染色见隐球菌,其中3例培养及涂片均为阳性。临床表现以头痛、发热、呕吐症状居多,1例并发癫痫,1例并发意识障碍。所有患者分别给予氟康唑、两性霉素B、氟胞嘧啶针、伏立康唑等抗真菌治疗,其中3例隐球菌脑膜炎患者予两性霉素B鞘内注射。经1~6个月治疗后,总体预后情况良好(好转22例,死亡1例)。结论肝、肾移植术后患者因免疫抑制剂的长期使用,隐球菌感染值得重视,其临床症状不典型,易误诊及漏诊,通过对其主要症状及影像学特点判断,结合肺穿刺活检、脑脊液检查、血培养等检查手段,可明显提高隐球菌感染的检出率,从而做到早诊断,早治疗,降低病死率。     

以皮肤受累为首发表现的播散性念珠菌病1例并文献复习

目的：播散性念珠菌病是一种致命性真菌感染性疾病,在免疫缺陷患者中发病率逐年增多,报道1例以双下肢多发皮下结节为首发表现,伴有肺及脑受累的播散性念珠菌病,并文献复习播散性念珠菌病的皮肤受累临床表现。方法患者女,37岁。因双下肢多发皮下结节6个月余就诊。有局灶节段性肾小球硬化病史,口服强的松及他克莫司2a余。取患者皮损组织行病理学检查,皮损组织、脓液、血、痰、尿、粪、脑脊液进行真菌镜检及真菌培养,并文献检索统计播散性念珠菌病皮肤受累患者临床特点。结果皮损组织病理见假菌丝,皮损组织、脓液、痰、尿、粪标本直接涂片均见假菌丝并培养出白念珠菌,CT显示肺受累,诊断为播散性念珠菌病,予抗真菌治疗,患者皮损愈合及肺部病灶部分吸收,但因自行停药,最终出现颅内播散。结论以皮损为首发表现的播散性念珠菌病临床罕见,临床诊疗中应重视应用免疫抑制剂患者皮损的组织病理及微生物检查,及早进行诊断和治疗,防止出现系统性播散,从而降低死亡率。     

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.     

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。     

热带念珠菌引起唇部念珠菌性肉芽肿1例

报告1例由热带念珠菌引起的唇部感染。患者女,22岁,下唇部斑块1 a。皮损组织病理学检查及真菌培养见热带念珠菌生长。给予伊曲康唑200 mg/d口服等治疗1个月后皮损完全消失。     

Degradation of phenol by aerobic granules and isolated yeast Candida tropicalis

Aerobic granules effectively degrade phenol at high concentrations. This work cultivated aerobic granules that can degrade phenol at a constant rate of 49 mg-phenol/g x VSS/h up to 1,000 mg/L of phenol. Fluorescent staining and confocal laser scanning microscopy (CLSM) tests demonstrated that an active biomass was accumulated at the granule outer layer. A strain with maximum ability to degrade phenol and a high tolerance to phenol toxicity isolated from the granules was identified as Candida tropicalis via 18S rRNA sequencing. This strain degrades phenol at a maximum rate of 390 mg-phenol/g x VSS/h at pH 6 and 30 degrees C, whereas inhibitory effects existed at concentrations >1,000 mg/L. The Haldane kinetic model elucidates the growth and phenol biodegradation kinetics of the C. tropicalis. The fluorescence in situ hybridization (FISH) and CLSM test suggested that the Candida strain was primarily distributed throughout the surface layer of granule; hence, achieving a near constant reaction rate over a wide range of phenol concentration. The mass transfer barrier provided by granule matrix did not determine the reaction rates for the present phenol-degrading granule.     

利用可重复使用的URA3标记基因建立热带假丝酵母基因敲除系统

热带假丝酵母(Candida tropicalis)在发酵工业中具有重要的应用潜力,但二倍体遗传结构和较低的遗传转化效率限制了其代谢工程育种技术的应用。建立可靠的遗传转化技术并高效的删除目的基因是代谢工程改造热带假丝酵母的重要前提。文章以C. tropicalis ATCC 20336为出发菌株,通过化学诱变筛选获得了尿嘧啶缺陷型突变株C. tropicalis XZX(ura3/ura3)。以丙酮酸脱羧酶(Pyruvate decarboxylase,PDC)基因作为靶基因构建了两端包含同源臂并在选择性标记C. tropicalis URA3(Orotidine-5′-phosphate decarboxylase,乳清酸核苷-5-磷酸脱羧酶)基因两侧同向插入源于沙门氏菌(Salmonella typhimurium)的hisG序列的基因敲除盒PDC1-hisG-URA3-hisG- PDC1(PHUHP),并转化宿主菌株C. tropicalis XZX,筛选获得PHUHP片段正确整合到染色体的PDC基因位点的转化子XZX02。在此基础上,将转化子XZX02涂布于5-FOA(5-氟乳清酸)选择培养基上,筛选得到URA3基因从PHUHP片段中丢失的营养缺陷型菌株XZX03。进一步构建了第2个PDC等位基因的删除表达盒PDCm- URA3-PDCm,并转化C. tropicalis XZX03菌株,获得转化子C. tropicalis XZX04。经PCR和DNA测序确认转化子C. tropicalis XZX04细胞染色体上的两个PDC等位基因被成功敲除。文章建立了一种营养缺陷型标记可重复使用的热带假丝酵母遗传转化技术,利用该技术成功敲除了细胞的PDC基因,为进一步利用代谢工程改造热带假丝酵母奠定了基础。     

A PCR-based approach to sequence the Candida tropicalis HSP90 gene

The heat shock protein 90 (hsp90) gene sequence is known to be highly conserved across the species barrier. A PCR-based method was thus utilised in an attempt to sequence the Candida tropicalis hsp90 gene. Primers for PCR were designed from conserved regions of the gene, which were identified by comparing the Saccharomyces cerevisiae and Candida albicans hsp90 gene sequences. Different sets of primers were designed to amplify and obtain overlapping DNA sequences of the C tropicalis gene. PCR was carried out on genomic DNA of Candidca tropicalis and the PCR products were cloned into suitable vector molecules for sequencing. In this way, a 2,070-basepair sequence of the C. tropicalis hsp90 gene was obtained. The PCR-based approach proved to be an easier method of obtain the sequence of a highly conserved gene, as compared to more conventional methods.     

1例热带念珠菌脓肿误诊分析

报告1例热带念珠菌引起的腹腔脓肿瘘管真菌感染。患者女,78岁,因十二指肠球部穿孔并发腹膜炎,行腹腔镜术,术后伤口反复未完全愈合,有少量分泌物溢出。发现腹壁瘘管及腹腔脓肿入院,两次细菌培养均无菌生长,再次取分泌物直接镜检阳性,真菌培养鉴定为热带念珠菌。手术清理组织,病理示感染性肉芽肿改变,查见大量假菌丝及孢子。经口服伊曲康唑和两性霉素B溶液纱条引流治疗3个月后基本痊愈。     

Salt-tolerant phenol-degrading microorganisms isolated from Amazonian soil samples

Two phenol-degrading microorganisms were isolated from Amazonian rain forest soil samples after enrichment in the presence of phenol and a high salt concentration. The yeast Candida tropicalis and the bacterium Alcaligenes faecoalis were identified using several techniques, including staining, morphological observation and biochemical tests, fatty acid profiles and 16S/18S rRNA sequencing. Both isolates, A. faecalis and C. tropicalis, were used in phenol degradation assays, with Rhodococcus erythropolis as a reference phenol-degrading bacterium, and compared to microbial populations from wastewater samples collected from phenol-contaminated environments. C. tropicalis tolerated higher concentrations of phenol and salt (16 mM and 15%, respectively) than A. faecalis (12 mM and 5.6%). The yeast also tolerated a wider pH range (3-9) during phenol degradation than A. faecalis (pH 7-9). Phenol degradation was repressed in C. tropicalis by acetate and glucose, but not by lactate. Glucose and acetate had little effect, while lactate stimulated phenol degradation in A. faecalis. To our knowledge, these soils had never been contaminated with man-made phenolic compounds and this is the first report of phenol-degrading microorganisms from Amazonian forest soil samples. The results support the idea that natural uncontaminated environments contain sufficient genetic diversity to make them valid choices for the isolation of microorganisms useful in bioremediation.