Genome Organization and Expression of the <Emphasis Type="Italic">Penicillium stoloniferum</Emphasis> Virus F

The complete sequences of three double-stranded (ds) RNAs (referred to F1, F2 and F3) of Penicillium stoloniferum virus F (PsV-F) were established. The F1 dsRNA was 1677 bp in length, and it contained one open reading frame (ORF) of 538 amino acids (molecular weight of 63 kDa, referred to P63), The F2 dsRNA was 1500 by in length, and also it contained one ORF of 420 amino acids (molecular weight of 46 kDa, referred to P46). The F3 dsRNA was 677 bp in length, but contained a small ORF with unknown function. A sequence motif of (5′-CGTAAAA-3′) was found only at the 5′ termini of the F1 and F2 dsRNAs, and a sequence motif of (5′-TAAAAAAAAA-3′) was found at the 3′ termini of all three dsRNA segments. The predicted amino acid sequence of F1 showed 38–48% sequence homology with the putative dsRNA-dependent RNA polymerases (RdRp) of dsRNA viruses, but the predicted amino acid of F2 showed no homology. Phylogenetic analysis using the RdRp sequences of the various Partitiviruses and Alphacryptoviruses revealed that PsV-F clustered well with Partitiviruses, but showed remote relationship with PsV-S. Near full-length and positive-sense single-stranded (ss) RNAs derived from the Fl, F2 and F3 dsRNAs were detected from the PsV-infected host cell. The expressed proteins of P63 and P46 showed a positive reaction against PsV-F antiserum, indicating P63 and P46 as RdRp and capsid protein, respectively. These results suggest that PsV-F can be a member of Partitivirus, but it is quite distinct from PsV-S electrophoretically, serologically and genetically, though both viruses coexist in the same cell.     

AST高于正常值上限2倍且大于ALT对AIDS合并青霉菌病的筛查价值

目的评价拟诊标准(血清AST水平高于正常值上限2倍且大于ALT)对AIDS合并马尔尼菲青霉菌病(penicilliosis marneffei,PSM)的筛查价值。方法对广西地区1045例疑似AIDS合并PSM的患者同时进行"金标准"(血、骨髓、组织培养或组织病理发现马尔尼菲青霉菌)及拟诊标准所需检查,评价拟诊标准对AIDS合并PSM的诊断价值。结果拟诊标准的灵敏度为45.79%、假阴性率42.11%、特异度90.64%、假阳性率9.36%、准确度82.49%、阳性似然比4.89、阴性似然比0.60、阳性预测值52.10%、阴性预测值88.27%。结论 AIDS患者如出现血清AST水平高于正常值上限2倍且大于ALT时,尚不能作为筛查PSM的常规指标,但应高度警惕PSM。     

艾滋病合并马尔尼菲青霉菌感染二例的病理特点

目的了解艾滋病合并马尔尼菲青霉菌感染的临床表现及组织病理学特点，探讨马尔尼菲青霉菌病的诊断及鉴别诊断。方法对2例活检标本进行常规苏木精一伊红染色及过碘酸一希夫染色、Gomori六胺银特殊染色，观察马尔尼菲青霉菌在组织中的形态特点，并结合文献进行讨论。结果在组织内可见酵母状真菌，过碘酸一希夫染色、Gomori六胺银真菌染色阳性，可见呈腊肠样结构并具有横隔。结论组织病理学检查，马尔尼菲青霉菌具有较典型的形态结构。     

艾滋病合并马尔尼菲青霉病感染率调查和实验室检查及药敏分析

目的探究艾滋病（AIDS）合并马尔尼菲青霉病（PSM）的感染率,以及常规实验室检查结果和5种抗真菌药物对马尔尼菲青霉菌（PM）的体外抗菌活性。方法 2006年1月2009年11月间确诊AIDS患者326例,从其血液、骨髓培养出65株PM,检测该65例患者的血常规、肝功能和肾功能,并对20株酵母相PM进行体外药敏进行分析。结果 AIDS合并PSM的感染率为19.94%;外周血常规：WBC〈4.0×109/L者48例,HB〈100g/L者51例,PLT〈100×109/L者46例。肝功能检查：ALT〉40U/L者49例,AST〉40U/L者51例,GGT〉60U/L者44例,ALP〉150U/L者36例,ALB〈35g/L者53例,A/G倒置者50例。肾功能检查：BUN〉7.2mmol/L者9例,Cr〉150μmol/L者4例。20株酵母相PM对5-氟胞嘧啶（5FC）、两性霉素B（AMB）、氟康唑（FCA）、伊曲康唑（ITR）、伏立康唑（VRC）的敏感率分别为75%、90%、80%、90%、90%。结论 AIDS合并PSM感染率较高;患者感染后外周血WBC、HB、PLT通常低于正常人,肝功能多表现异常,肾功能的改变较少;对PM的治疗以AMB、ITR、VRC为首选。     

PAS联合六胺银染色诊断局限性马尔尼菲青霉菌感染的应用

目的探讨建立快速诊断局限性马尔尼菲青霉菌感染的方法,以期早期诊断,降低死亡率。方法采用高碘酸一无色品红（PAS）染色联合六胺银染色对4例非AIDS马尔尼菲青霉菌感染者活检组织进行马尔尼菲青霉菌的特殊染色,分析感染者的临床表现和病理组织学诊断特点。结果患者均为广东本地居民,属局限性马尔尼菲青霉菌感染。除1例有发热外,临床上未见以发热、失重、贫血、白细胞增高为主的马尔尼菲青霉菌感染症状以及多器官损害症状。HE染色显示病理组织学类型为肉芽肿型和化脓型。病变组织经PAS染色与六胺银染色后,在油镜下增生的泡沫状巨噬细胞胞浆内,呈现大量红色或黑色的卵圆形及腊肠状酵母样青霉菌孢子。无荚膜,在腊肠状菌体中可见马尔尼菲青霉菌特有的裂殖时出现的横隔膜。结论PAS联合六胺银染色清晰显示组织中马尔尼菲青霉菌的特征性结构和形态,结合病理组织学特点、中国南方地区生活或旅游史,可以快速诊断局限性马尔尼菲青霉菌感染。     

广泛播散型马尔尼菲青霉病患者的护理

报告了1例广泛播散型马尔尼菲青霉病患者的护理。主要内容包括:心理护理、皮肤护理、药物不良反应的观察、发热护理、营养护理、标本采集及配合诊断、控制感染、严密观察病情等。经过近3个月的精心治疗与护理,患者康复出院。     

艾滋病合并马尔尼菲青霉菌感染的护理

目的探讨艾滋病合并马尔尼菲青霉菌感染患者的护理及体会。方法通过临床诊断,对66例艾滋病合并马尔尼菲青霉菌感染的患者实施药物治疗并结合药物护理、心理护理等方法,观察患者临床症状的改善情况。结果66例患者中8例症状改善后放弃治疗,有13例由于入院时病情危重,合并多种耐药菌感染,多器官功能衰竭死亡,其余好转。结论对艾滋病患者合并马尔尼菲青霉菌感染,药物护理和心理护理具有重要的意义。     

Integrative transformation of the mycotoxin-producing fungus,Penicillium paxilli

A high frequency transformation system has been developed for Penicillium paxilli using pAN7-1. Up to 44% of the primary transformants were heterokaryons. Loss of hygromycin resistance was observed in primary transformants that were sub-cultured on non-selective media, but single spores of these primary transformants were mitotically stable on both selective and non-selective media. A molecular analysis of the transformants generated showed that 78% had single-site integrations, with half of these containing a single copy of pAN7-1. CHEF-gel electrophoresis showed that P. paxilli has at least six chromosomes with a total genome size of about 23.4 Mb.