Replication-independent expression of genome components and capsid protein of brome mosaic virus in planta: a functional role for viral replicase in RNA packaging

To begin elucidation of the relationship between Brome mosaic virus (BMV) replication and encapsidation, we used a T-DNA-based Agrobacterium-mediated transient expression (agroinfiltration) system in Nicotiana benthamiana leaves to express either individual or desired pairs of the three genomic RNAs. The packaging competence of these RNAs into virions formed by the transiently expressed coat protein (CP) was analyzed. We found that in the absence of a functional replicase, assembled virions contained non-replicating viral RNAs (RNA1 or RNA2 or RNA3 or RNA1 + RNA3 or RNA2 + RNA3) as well as cellular RNAs. By contrast, virions assembled in the presence of a functional replicase contained only viral RNAs. To further elucidate the specificity exhibited by the functional viral replicase in RNA packaging, replication-defective RNA1 and RNA2 were constructed by deleting the 3' tRNA-like structure (3' TLS). Co-expression of TLS-less RNA1 and RNA2 with wt RNA3 resulted in efficient synthesis of subgenomic RNA4. Virions recovered from leaves co-expressing TLS-less RNA1 and RNA2 and either CP mRNA or wt RNA3 exclusively contained viral RNAs. These results demonstrated that packaging of BMV genomic RNAs is not replication dependent whereas expression of a functional viral replicase plays an active role in increasing specificity of RNA packaging.     

临床病毒性脑炎标本的博尔纳病病毒核酸检测

目的 探讨临床病毒性脑炎(viral encephalitis,VE)患者与博尔纳病病毒(Borna disease virus,BDV)感染的关系,分析BDV感染的病毒性脑炎患者临床特征.方法 用荧光定量巢式逆转录聚合酶链反应(FQ-nRT-PCR)方法检测病毒性脑炎患者及非感染性疾病施行蛛网膜下腔阻滞麻醉的手术患者脑脊液单个核细胞(cerebrospinal fluid mononuclear cell,CSFMC)中BDV p24基因片段,同时用β-肌动蛋白(β-actin)作为内参照,脑脊液(CSF)阳性标本基因测序分析并总结出临床特征.结果 32例病毒性脑炎脑脊液标本BDV p24基因片段检出率为12.5%(4/32),拷贝数＞102/μl.对其中一份CSF阳性标本测序后,与BDV标准病毒株Ⅴ和马源的BDV病毒株H1766序列比较同源性分别为95.35%和98.84%.在4个位点出现基因突变(nt1649 T→C、nt1656 G→A、nt1670 C→T、nt1676 C→T).该目的基因片段与马源的BDV病毒株亲缘关系最近.阳性脑炎患者主要以精神行为异常为临床特征.结论 贵州省遵义市部分病毒性脑炎的发生与BDV感染有关,主要以精神行为异常为临床特征.     

重型肝炎中甲、乙型肝炎病毒合并感染的探讨

报告经血清免疫学抗HAVIgM、抗-HBcIgM、HBV-DNA、HBsAg/IgM复合物以及乙肝三种抗原抗体系统检测48例重型肝炎中甲、乙型肝炎病毒感染情况。结果:31例为HBV感染,1例为HAV感染,1例未定型,15例(31.25％)为甲、乙型合并感染(混合感染7例、重叠感染8例)。15例中亚急性10例、慢性5例;死亡或恶化10例,与同期“单纯”感染的重肝比较,包括主要临床生化指标均无显著差异。有7例患者病程中有近期出现“急黄肝”症状的历史,随后病情突然加重;1例发生在住院期间,且血清抗-HAVIgM转阳,最后死亡。认为合并感染可使部分急、慢性肝炎病情加重,甚至发展成重型。应注意发现。     

乙型肝炎病毒相关肾炎临床与病毒血清学特点关联研究

目的 研究乙型肝炎病毒（HBV）感染与肾小球肾炎的关系,探讨HBV复制在HBV相关肾炎发病中的作用。方法 对338例经肾穿刺证实的肾炎患者的临床病理和病毒血清学特点进行分析。结果 HBV相关肾炎占肾炎总数的5．3％。在血HBsAg( )的肾炎患者中,HBV相关肾炎发病与HBeAg相关（P＜0．05）,与血清HBV DNA含量（bDNA)显著相关（P＜0．01）。分支链DNA（bDNA)阳性率明显高于HBeAg阳性率（P＜0．05）。结论 bDNA是反映HBV复制的最佳指标。HBV相关肾炎与体内HBV复制程度密切相关,HBV在体内大量复制,可能通过循环中的病毒抗原沉积于肾脏和肾脏中该抗原原位表达而致病。     

Third-generation smallpox vaccine strain-based recombinant vaccines for viral hemorrhagic fevers

Vaccinia virus has been used as a smallpox vaccine. Now that smallpox has been eradicated, the vaccinia virus is expected to be used as a bioterrorism countermeasure and a recombinant vaccine vector for other infectious diseases, such as viral hemorrhagic fevers. Many vaccinia virus strains were used as smallpox vaccines in the smallpox eradication campaign coordinated by the World Health Organization. These strains can be classified into generations, according to the history of improving production methods and efforts to reduce the adverse reactions. Significantly, the third-generation of smallpox vaccine strains, which include modified vaccinia Ankara (MVA) and LC16m8, are currently popular as recombinant vaccine vectors due to their well-balanced safety and immunogenicity profiles. The present review firstly focuses on the characteristics of the smallpox vaccine generations. The historical background of the development of the third-generation smallpox vaccine strains is detailed, along with the history of the transition of the vaccinia virus generation used as vectors for hemorrhagic fever vaccines to the third generation. Among the vaccinia viruses, MVA is currently the most commonly used vector for developing hemorrhagic fever vaccines, including dengue fever, yellow fever, Ebola viral disease, Lassa fever, Rift Valley fever, and Crimean-Congo hemorrhagic fever. LC16m8 is a vaccine candidate for severe fever with thrombocytopenia syndrome. The current status and recent advances in the development of these hemorrhagic fever vaccines using third-generation vaccinia strains are discussed.     

Applications of platform technologies in veterinary vaccinology and the benefits for one health

The animal-human interface has played a central role in advances made in vaccinology for the past two centuries. Many traditional veterinary vaccines were developed by growing, attenuating, inactivating and fractioning the pathogen of interest. While such approaches have been very successful, we have reached a point where they have largely been exhausted and alternative approaches are required. Furthermore, although subunit vaccines have enhanced safety profiles and created opportunities for combined discrimination between vaccinated and infected animal (DIVA) approaches, their functionality has largely been limited to diseases that can be controlled by humoral immunity until very recently. We now have a new generation of adjuvants and delivery systems that can elicit CD4 + T cells and/or CD8 +  T cell responses in addition to high-titre antibody responses. We review the current vaccine platform technologies, describe their roles in veterinary vaccinology and discuss how knowledge of their mode of action allows informed decisions on their deployment with wider benefits for One Health.     

病毒性肝炎患者血清IL-6和T细胞亚群的关系

采用ＭＴＴ比色法和抗体致敏的红细胞花环试验（间接法）　检测了９１例病毒性肝炎患者血清ＩＬ－６和Ｔ细胞亚群。　结果显示，病毒性肝炎患者血清ＩＬ－６活性均明显高于正常对照组（Ｐ＜０．０１），以重型肝炎为最高；且与肝细胞损害程度呈正比（Ｐ＜０．０１）；ＣＤ４~＋细胞、ＣＤ４＋／ＣＤ８~＋均明显降低，与血清ＩＬ－６呈负相关；ＣＯ８~＋细胞明显增高，与血清ＩＬ－６呈正相关（Ｐ＜０．０１）。表明，血清ＩＬ－６与机体细胞免疫功能密切相关，二者互为因果，共同参与病毒性肝炎的免疫病理损害，可作为判断病情和预后及免疫调节治疗的监测指标。     

北京地区病毒性脑炎患者病原体的分离和初步鉴定

１９９１年夏秋季，我们从４１例北京儿童医院临床诊断为病毒性脑炎（非乙脑）患者的７５份急性期血和／或脑脊液中分离到３３株病毒。对其中１２株进行电镜观察，电镜下均见大小相同的球形病毒颗粒，毒粒的直径约为４４．７８±１．３５ｕｍ，无囊膜，具双层蛋白外壳，视野内多见空心颗粒。超薄切片显示病毒在感染细胞的胞浆内发生，直径约为４７．６±２．３ｕｍ。正常细胞中未见病毒颗粒。对其中３株病毒进行理化性状分析，多次试验均显示该病毒抵抗５＇－碘脱氧尿苷，抵抗乙醚，耐酸，能在地鼠肾ＢＨＫ－２１和白纹伊蚊Ｃ６／３６细胞上增殖并出现细胞病变。该病毒与本室制备的披膜病毒科甲组、乙脑病毒和布尼亚病毒科的组特异性免疫腹水均不反应。脑内接种３日龄乳小白鼠可引起不规律的发病和死亡。上述结果表明，该病毒是一类４５ｕｍ左右、无囊膜、耐酸的ＲＮＡ病毒。     

小儿喉乳头状瘤HPV-DNA及体液免疫检测

目的：探讨小儿喉乳头状瘤（ＪＬＰ）人乳头瘤病毒（ＨＰＶ）感染途径及发病机理。方法：采用ＰＣＲ及ＰＣＲ产物斑点杂交技术检测ＪＬＰ组织ＨＰＶ－ＤＮＡ;散射免疫比浊法测定血清Ｉｇ及补体Ｃ３。结果：ＪＬＰ组织ＨＰＶ总感染率为９５％（１９／２０）,其中ＨＰＶ。型为５５％（１１／２０）,ＨＰＶ１１为３０％（６／２０）,ＨＰＶ６＋１１型为１０％（２／２０）;ＪＬＰ患者血清ＩｇＧ、ＩｇＡ、ＩｇＭ、Ｃ３值正常,对照     

诱导型一氧化氮合酶在实验性病毒性心肌炎心肌内的表达及其与心肌损害的关系

目的 探讨诱导型一氧化氮合酶（ｉＮＯＳ）在急性病毒性心肌炎心肌内的表达及其与心肌损害的关系。方法 ６～８周龄ＢＡＬＢ／Ｃ雄性小鼠２４只,随机分两组：感染组１６只以柯萨奇病毒Ｂ３ｍ（ＣＶＢ３ｍ）腹腔内注射,另８只为正常对照组。采用免疫组化法检测两组小鼠心肌组织内ｉＮＯＳ表达,并将其半定量与心肌病理变化程度进行相关分析,用比色法测定了血清中ＮＯ间接含量。结果 ①感染组小鼠心肌标本病理组织学检查均有不同