934株自血培养分离致病菌的病原学分析

目的了解中山大学附属第一医院2002年到2005年自血培养标本中分离的致病细菌的耐药状况和菌群分布特征。方法采用全自动血培养仪进行培养,血培养报阳后转种血平板,用VITEK-60鉴定所分离致病菌,药敏试验用K-B法。结果4年间血培养共分离出934株病原细菌,其中大肠埃希菌、金黄色葡萄球菌、肺炎克雷伯菌,是该院引起血液感染的主要致病细菌。药物敏感试验显示革兰阴性杆菌对亚胺培南、哌拉西林/他唑巴坦、头孢哌酮/舒巴坦的敏感性最高,而革兰阳性球菌对万古霉素及替考拉宁的敏感性最高。结论血液感染患者的死亡率较高,应重视导致血液感染致病菌的病原学研究,以利于疾病的治疗。     

颅内感染患者细菌分布及耐药性分析

目的分析神经外科颅内感染感染患者细菌的分布及其耐药性,指导合理应用抗生素及感染管理。方法回顾分析2009年至2010年神经外科颅内感染患者的细菌分离株及耐药性。结果细菌共92株,革兰阳性球菌40株,革兰阴性杆菌52株;排前6位的病原菌分别是鲍曼不动杆菌(15.22%)、表皮葡萄球菌(13.04%)、金黄色葡萄球菌(10.87%)、铜绿假单胞菌(10.87%)、肺炎克雷伯菌(8.7%)和粘质沙雷菌(8.7%)。结论神经外科颅内感染中革兰阴性菌与阳性菌比例相当,多重耐药性比例高;依据细菌及耐药性监测结果指导抗生素的合理应用,是治疗颅内感染的重要手段。     

拮抗菌株Kc-t99的鉴定及其抑菌活性研究

通过传统分类与分子生物学技术相结合的方法对从京郊菜园土壤中分离筛选的拮抗菌株Kc-t99进行鉴定,并采用生物测定的方法评价其抑菌活性.结果表明,菌株Kc-t99的形态学特征和生理生化特性与枯草芽孢杆菌基本一致,其16S rDNA序列与GenBank中已鉴定的枯草芽孢杆菌的16S rDNA序列同源性高达98.06%,据此初步确定其为枯草芽孢杆菌.抑菌试验表明该菌株对供试的5种病原真菌和4种细菌均具抑菌活性,其中对甘蓝枯萎病菌、黄瓜角斑病菌和桃褐腐病菌抑制作用显著.     

实验动物皮肤病原真菌检测方法的改良

目的对实验动物皮肤病原真菌2种培养方法进行了比较。方法将采集到的3只皮肤真菌感染病兔样品经由沙氏平皿法和沙氏试管斜面培养法分别进行培养。结果在3只真菌感染病兔中应用试管斜面法我们只检测到1例皮肤病原真菌阳性,而采用沙氏平皿法3例阳性全部检出。结论结合临床检测经验,我们认为本研究的沙氏平皿法优于沙氏试管斜面法,在实验动物皮肤病原真菌常规检测中具有推广应用价值。     

儿童活体肝移植术后感染常见致病菌及其耐药性分析

目的分析儿童活体肝移植术后常见致病菌的分布特点及其对常用抗菌药物的耐药性情况,为合理使用抗生素提供参考。方法回顾性分析重庆医科大学附属儿童医院自2006年6月至2009年12月术后监护的42例儿童活体肝移植患儿送检共152份标本进行菌株鉴定及药敏试验。结果经培养共分离出66株致病菌,阳性率为43.4%,以呼吸道来源为主,占71.2%。66株致病菌中革兰阴性菌（G^-）63.6%,革兰阳性菌（G^＋）27.3%,真菌9.1%。常见G^-菌为铜绿假单胞菌、阴沟肠杆菌、大肠埃希菌和肺炎克雷伯菌肺炎亚种。常见G^＋菌为金黄葡萄球菌和肺炎链球菌。真菌主要为白色假丝酵母菌。药敏结果显示细菌耐药性明显增强,G^-菌中阴沟肠杆菌、大肠埃希菌和肺炎克雷伯菌肺炎亚种对美洛培南、亚胺培南100%敏感,对环丙沙星、左旋氧氟沙星、阿米卡星和氯霉素敏感在85.7%以上;超广谱β-内酰胺酶（ESBLs）菌株检出率为66.7%100%。铜绿假单胞菌仅对环丙沙星、左旋氧氟沙星、庆大霉素、阿米卡星和多粘菌素敏高度感度,敏感度在91.6%以上。G^＋菌中金黄色葡萄菌对万古霉素、替考拉宁、利奈唑胺、呋喃妥因、阿米卡星和环丙沙星100%敏感,β-内酰胺酶检出率为100%。肺炎链球菌对万古霉素、利奈唑胺、头孢西丁、美洛培南、左旋氧氟沙星和头孢吡肟均有83.3%以上的敏感性,对于阿莫西林仍有66.7%的敏感性,但对于大环内酯类100%耐药。白色假丝酵母菌对两性霉素B 100%敏感。结论儿童活体肝移植术后常见致病菌以G^-菌为主,多为多重耐药菌株,含有β-内酰胺酶抑制剂及碳青霉烯类抗生素是治疗该类细菌的有效抗生素。万古霉素是G^＋菌感染的有效抗生素。两性霉素B是真菌感染的有效药物。为避免耐药率上升,临床应合理使用抗生素。     

Crystal Structures of Trypanosomal Histidyl-tRNA Synthetase Illuminate Differences between Eukaryotic and Prokaryotic Homologs

Crystal structures of histidyl-tRNA synthetase (HisRS) from the eukaryotic parasites Trypanosoma brucei and Trypanosoma cruzi provide a first structural view of a eukaryotic form of this enzyme and reveal differences from bacterial homologs. HisRSs in general contain an extra domain inserted between conserved motifs 2 and 3 of the Class II aminoacyl-tRNA synthetase catalytic core. The current structures show that the three-dimensional topology of this domain is very different in bacterial and archaeal/eukaryotic forms of the enzyme. Comparison of apo and histidine-bound trypanosomal structures indicates substantial active-site rearrangement upon histidine binding but relatively little subsequent rearrangement after reaction of histidine with ATP to form the enzyme's first reaction product, histidyladenylate. The specific residues involved in forming the binding pocket for the adenine moiety differ substantially both from the previously characterized binding site in bacterial structures and from the homologous residues in human HisRSs. The essentiality of the single HisRS gene in T. brucei is shown by a severe depression of parasite growth rate that results from even partial suppression of expression by RNA interference.     

Acanthamoeba castellanii: Morphological analysis of the interaction with human cornea

The present study demonstrates that when Acanthamoeba castellanii trophozoites are co-cultivated with isolated human corneas, the amoeba can be invasive and cause damage to the intact corneal epithelium without the requirement of previous corneal abrasion. After adhesion, A. castellanii trophozoites migrate between cells forming bumps on the corneal cell layers and reaching Bowman´s membrane in 3 h, although no evidence of cell damage was observed until the phagocytic process was detected. Likewise, conditioned medium produced damage to the corneal cells that was proportional to the time of incubation, but this cytophatic effect involved only the most superficial layer of the human cornea and was not enough to explain amoebic invasion of Bowman´s membrane. As a result of our observations, we suggest that the mechanical action of the trophozoites and phagocytosis of corneal cells during the process of corneal invasion are more important than previously suggested.     

Toward resolving family-level relationships in rust fungi (Uredinales)

Rust fungi (Basidiomycota, Uredinales) consist of more than 7000 species of obligate plant pathogens that possess some of the most complex life cycles in the Eumycota. Traditionally, a limited number of synapomorphic characters and incomplete life-cycle and host-specificity data have hampered phylogenetic inference within the Uredinales. The application of modern molecular characters to rust systematics has been limited, and current contradictions, especially in the deeper nodes, have not yet been resolved. In this study, two nuclear rDNA genes (18S and 28S) were examined across the breadth of the Uredinales to resolve some systematic conflicts and provide a framework for further studies of the group. Three suborders of rusts are recovered. Of the 13 rust families most widely accepted, 8 are supported in full or in part (Coleosporiaceae, Melampsoraceae, Mikronegeriaceae, Phakopsoraceae p.p., Phragmidiaceae, Pileolariaceae, Pucciniaceae, Raveneliaceae), 3 are redundant (Cronartiaceae, Pucciniastraceae, Pucciniosiraceae), and the status of 2 (Chaconiaceae, Uropyxidaceae) could not be resolved. The Mikronegeriaceae and Caeoma torreyae are the most basal rusts sampled. It is concluded that morphology alone is a poor predictor of rust relationships at most levels. Host selection, on the other hand, has played a significant role in rust evolution. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the US Department of Agriculture.     

病原真菌感染与TOLL样受体

TOLL样受体(TLR)是参与天然免疫的主要模式识别受体之一,与许多微生物病原体及其产物的病原相关分子模式PAMP结合后通过MyD88依赖性或非依赖性途径启动宿主胞内信号传导途径,引发一系列生物学效应。白色念珠菌表面的特征性糖磷脂甘露聚糖可被TLR2、TLR4识别,诱导前炎性细胞因子的释放及促进中性粒细胞的聚集等来介导宿主的抗真菌免疫反应。烟曲霉则可能利用表型转换(酵母样与菌丝态),通过不同TLRs逃避宿主天然免疫系统的识别。新型隐球菌的多糖荚膜成分葡糖醛氧化甘露聚糖GXM可与TLR2、TLR4、CD14结合,在单核细胞、巨噬细胞对GXM的内化、吞噬中起重要作用,而不是诱导细胞因子的分泌;酿酒酵母胞壁成分酵母多糖则可激活TLR2、TLR6异源二聚体。总之,TLR与真菌配体相互作用的具体机制及其活化后胞内信号传导调控机制的深入研究与分析,对临床真菌病的免疫调节及治疗具有重要意义。