Platelet interactions with viruses and parasites

Abstract

While the interactions between Gram-positive bacteria and platelets have been well characterized, there is a paucity of data on the interaction between other pathogens and platelets. However, thrombocytopenia is a common feature with many infections especially viral hemorrhagic fever. The little available data on these interactions indicate a similarity with bacteria-platelet interactions with receptors such as FcγRIIa and Toll-Like Receptors (TLR) playing key roles with many pathogens. This review summarizes the known interactions between platelets and pathogens such as viruses, fungi and parasites.     

2016至2018年某医院真菌血流感染者流行病学特征及耐药性分析

目的分析真菌性血流感染的病原菌分布以及耐药特征，为真菌血流感染的早期合理用药提供理论依据。 方法回顾性分析武汉大学人民医院2016年1月至2018年12月收治的真菌性血流感染者的菌群、科室分布以及耐药性。 结果入组192例真菌血流感染者的血培养样本中共分离192株真菌，其中白色念珠菌检出率为31.77%（61/192），其次热带念珠菌检出率为18.75%（36/192）；重症医学科检出率最高为33.85%（65/192）。所有菌株均对两性霉素B敏感，对其他抗菌药物耐药率分别为5-氟胞嘧啶4.49%（9/192）、伊曲康唑5.73%（11/192）、氟康唑10.94%（21/192）和伏立康唑11.46%（22/192）；除两性霉素B外，2016至2018年真菌对其他抗菌药物的耐药率均逐年上升，其中2018年所分离192株光滑念珠菌对伊曲康唑耐药菌率达46.7%。 结论真菌血流感染病原菌以念珠菌属为主，对目前抗真菌药物具有较高敏感性，但耐药率逐年上升，加强监测血培养病原菌变化及耐药趋势对指导临床用药至关重要。     

广东省中山市头癣现状及其病原体分离结果分析

目的:了解广东省中山地区头癣现状及其病原体分布、流行情况。方法:对107例镜检阳性病发进行真菌培养、鉴定和临床分析。结果:107例患者中白癣81例(占75.7％),脓癣24例(22.4％),黑癣2例(1.9％),未发现黄癣。107例病发分离出5种共101个菌株,其中犬小孢子菌90株(占89.1％),须癣毛癣菌5株(4.9％),红色毛癣菌3株(2.79％),断发毛癣菌2株(1.98％),紫色毛癣菌1株(0.99％)。结论:广东省中山地区头癣种类以白癣为主,脓癣次之,黑癣少见,黄癣基本消失,流行的病原菌以亲动物性的犬小孢子菌占绝对多数。发病年龄以1岁～15岁的少年儿童为主,成年人少见。     

呼吸道真菌感染核酸胶体金试纸条快速检测方法的建立和评价

目的 采用聚合酶链式反应（PCR）技术和胶体金技术建立一种呼吸道真菌感染核酸胶体金试纸条检测方法,并对其检测效果进行评价。 方法 根据GenBank数据库选择真菌内转录间隔区（ITS）基因片段作为靶基因,应用DNAMAN软件对真菌ITS基因片段进行分析,并分别在真菌通用引物5'端用生物素（Biotin）和异硫氰酸荧光素（FITC）进行修饰标记;建立PCR体系,进行体系的最佳条件优化;确定胶体金免疫层析试纸条标记链霉亲和素的最佳标记量和质控线及检测线最佳包被物浓度,组装核酸胶体金试纸条,对试纸条的特异度、灵敏度、重复性和稳定性进行验证。 结果 水煮法提取白假丝酵母菌、新生隐球菌和毛霉菌DNA,浓度为180 μg·L^－1以上,纯度为1.60~2.10;在pH 7.0条件下,每100 μL胶体金溶液中制备胶体金标记链霉亲和素最佳标记量为 3.3 μg,质控线包被Biotin化牛血清白蛋白（BSA-Biotin）浓度为2.00 g·L^－1,检测线包被抗FITC抗体浓度为1.00 g·L^－1。核酸试纸条的特异度与电泳结果一致,仅白假丝酵母菌、新生隐球菌和毛霉菌出现阳性结果,与金黄色葡萄球菌、肺炎链球菌、乙型溶血性链球菌、铜绿假单胞菌、肺炎克雷伯菌、鲍曼不动杆菌和大肠埃希菌等常见呼吸道感染细菌无交叉反应。灵敏度检测,白假丝酵母菌、新生隐球菌和毛霉菌的DNA浓度分别为10^－4、10^－2和10^－3 mg·L^－1时核酸试纸条仍可准确检出,而普通PCR电泳结果显示白假丝酵母菌、新生隐球菌和毛霉菌最低检测浓度分别为0.01、1.00和0.10 mg·L^－1;重复性检测,在不同实验室不同操作人员对核酸试纸条进行验证,结果一致,重复性良好;稳定性检测,核酸试纸条在第6、9和12个月进行检测,结果符合预期,稳定性良好。 结论 本研究建立的呼吸道真菌感染核酸胶体金试纸条可以检测白假丝酵母菌、新生隐球菌和毛霉菌等真菌,特异度和灵敏度均较高,操作简便且快速。     

Ribosomal control of translational fidelity in Podospora anserina: A suppressor and an antisuppressor affecting the paromomycin-induced misreading in vitro

Summary The hypersensitivity to paromomycin dis played by the su _1–60 suppressor, which was thought to be a ribosomal ambiguity mutation, was used to isolate a new antisuppressor, AS ₇₁. This antisuppressor is not specific for ribosomal suppressors, since it is able to act on putative tRNA-type suppressors. Compared to the wild-type, the AS _7-1 mutation produces an increased resistance to paromomycin. In addition, it was found to stop the sporulation process. In the polyuridylic acid-dependant assay, the paromomycin induced misreading was shown to be higher on su _1–60 ribosomes and lower on ribosomes from AS _7–1 mutant and AS _7S-1-su _1–60 double mutant than on wild-type ribosomes. These data confirm that su _1–60 and AS _7-1 mutants are ribosomal mutants involved in the control of translational fidelity.     

基因芯片技术检测鉴定临床常见致病真菌的初步研究

目的为了快速、简便、高通量地鉴定临床常见致病真菌,建立了一种采用基因芯片技术对临床常见的致病真菌鉴定的分子生物学方法。方法以5.8S rDNA与28S rDNA间的内转录间区2(internal transcribed spacer-2,ITS-2)为靶标,针对待检的临床常见致病真菌设计合成一系列寡核苷酸探针,制成寡核苷酸芯片。待检真菌DNA经通用引物扩增标记后,与芯片杂交,对杂交图谱分析归纳,得到一套种特异性的典型杂交图谱。待检的样品菌与基因芯片杂交,得到的杂交结果与典型图谱比对即可判断出样品的种类。结果以涉及8个属20个种的标准致病真菌菌株对芯片的特异性、重复性、灵敏度进行考察,结果表明,该研究建立的基因芯片技术可以有效地区分20种临床常见致病真菌,特异性良好,重复性良好(信噪比CV<10%),灵敏度为15 pg/ml真菌DNA。收集从临床分离的84株致病真菌菌株对基因芯片进行试用,结果显示基因芯片的鉴定结果与常规鉴定方法的鉴定结果一致。结论这项技术的建立可以稳定、特异性地实现临床常见致病真菌的高通量鉴定,为进一步检测研究奠定了基础。     

Recombinational stability of replicating plasmids in Aspergillus nidulans during transformation,vegetative growth and sexual reproduction

Plasmids containing the AMA1 replicon are capable of autonomous maintenance in Aspergillus nidulans. It has been reported previously that these plasmids can form concatenates by recombination in a transformed mycelium, and up to 10% of molecules are involved in such events. The present study demonstrates that plasmid recombination, although frequent during transformation, rarely occurs during vegetative growth. As a result, the structure and phenotypic stability of AMA1 plasmids generally remains unaltered for many asexual (conidial) generations. It is also evident that plasmid replication does not require specific recombination events in the AMA1 palindrome. However, during sexual reproduction, autonomous plasmids exhibit increased recombination, which results in both plasmid concatenation and integration into the chromosome.     

Transformation of the cultivated mushroom Agaricus bisporus (Lange) using T-DNA from Agrobacterium tumefaciens

Agrobacterium tumefaciens is known to transfer parts of its tumor-inducing plasmid, the T-DNA, to plants, yeasts and filamentous fungi. We have used this system to transform germinating basidiospores and vegetative mycelium of a commercial strain of the cultivated basidiomycete Agaricus bisporus. Analysis of transformants shows that the T-DNA integrates at random sites into the host genome and that the selection marker is stable during mitosis and meiosis. The Agrobacterium system allows the transformation of both homokaryons and heterokaryons of A. bisporus. Also, both karyotypes of an heterokaryon can be transformed simultaneously. Furthermore, this is the first report on the transformation of vegetative mycelium of a commercial strain of A. bisporus. Received: 6 June 2000 / Accepted: 10 October 2000     

急性爆发性真菌性鼻窦炎

目的　探讨急性爆发性真菌性鼻窦炎 (acutefulminantfungalsinusitis ,AFFS)的临床特征、诊断标准及治疗原则。方法　回顾收治的 6例较典型的AFFS病例诊断和治疗过程。其中结合全身疾病病史 ,发热伴颜面部、眼部及鼻部症状 ,影像学 ,鼻内镜及鼻腔分泌物真菌涂片检查 ,在患者就诊的 2 4h内做出初步诊断 5例。在此基础上鼻内镜下行全组鼻窦开放术 ,切除全部坏死组织至露出新鲜创面 5例。根据临床及影像学表现 ,进行眶内容物切除 1例。术后同时进行原发病治疗以及全身抗真菌治疗 5例。结果　 6例病例经过病理、真菌涂片及培养证实为鼻窦黏膜侵袭性真菌感染 ,其中毛霉菌 1例 ,根霉菌 2例 ,链隔孢霉菌 1例 ,曲霉菌 1例 ,毛霉菌根霉菌混合 1例。 1例未在第一时间诊断 ,且未经抗真菌治疗的患者于住院第 7天死亡 ,1例抗真菌治疗及清创术后 88d死亡 ,2例分别于治疗后 3 2个月和 6个月死于白血病 ,2例经上述治疗后分别随访 9个月和 11个月无复发 ,后者为本组唯一进行眶内容物摘除者。结论　根据病史 ,眼或颜面症状 ,鼻腔内干痂以及影像学尤其是磁共振成像 ,分泌物真菌涂片检查 ,可以在患者到鼻科就诊的 2 4h内做出AFFS的诊断。包括及时彻底的根治性清创 ,足量静脉内二性霉素B注射 ,完全控制原发病以及足够的全身支     

Molekulare Aspekte und diagnostische Wertigkeiten von Pilzallergenen

Zusammenfassung. Die Klonierung, Sequenzierung und Produktion von hochreinen Allergenen bietet die Möglichkeit, perfekt standardisierte Allergenpräparate herzustellen. Die Entwicklung eines neuen Klonierungssystems, das auf filamentösen Phagen basiert, führte zu einer schnellen Isolierung und Charakterisierung von Aspergillus fumigatus-Allergenen. Die auf diesem Weg rekombinant hergestellten Proteine wurden serologisch und klinisch geprüft und ihr routinemä-ßiger Einsatz im ImmunoCAP-System evaluiert. Es gelang eine quantitative Übereinstimmung zwischen Hauttestergebnissen und Serologie nachzuweisen, welche das Potential rekombinanter Allergene in der Diagnostik allergischer Krankheiten aufzeigt. Darüber hinaus trägt die Charakterisierung der Pilzallergene wesentlich zum Verständnis der moiekularen Natur der allergieauslösenden Komponenten bei Zum jetzigen Zeitpunkt können, abgesehen von Proteinen mit unbekannten biologischen Funktionen, die Pilzallergene in zwei Klassen eingeteilt werden: 1. Spezies-spezifische sezcrnierte Allergene und 2. cytoplasmatische, hoch konservierte Proteine. Diese letztgenannten Pilzallergene zeigen auch zu Proteinen aus phylogenetisch weit entfernten Organismen weitreichende Sequenzhomologien. Neben der daraus zu erwartenden IgE-Kreuzreaktivität findet man in einigen Fällen auch eine Kreuzreaktivität mit den homologen humanen Proteinen, was auf Autoimmunreaktionen, bei Pilzalleigien hindeutet. Summary. Cloning, sequencing and production of highly pure recombinant allergens allows to produce perfectly standardised allergen preparations. The development of a new cloning system based on filamentous phage allowed the fast isolation and characterisation of allergens from the fungus Aspergillus fumigatus. The produced recombinant allergens were tested in serological and clinical studies as well as for their performance for routine assessments in the ImmunoCAP-system. Thereby, a perfect correlation between skin test results and serology was found showing the potential of recombinant allergens for the diagnosis of allergic diseases. Moreover, the characterisation of fungal allergens substantially contributes to our understanding of the molecular nature of proteins involved in the elication of allergic reactions. Apart from allergenic proteins with unknown biological function, fungal allergens can be subdivided into two classes: 1. Species-specific, secreted proteins and 2. cytoplasmic, even in phylogenetically distant organisms, well conserved proteins. These fungal allergens show extended sequence similarity, a high level of IgE cross-reactivity and in some cases also cross-reactivity with homologous human proteins indicating autoimmune reactions involved in fungal allergy.