猪伪狂犬病病毒抗体检测的研究进展

伪狂犬病(PR)又名Aujeszky病,是OIE规定的通报疫病,由疱疹病毒引起的严重传染病,具有传播速度快、传播途径多和流行范围广等特点,成为仅次于口蹄疫和猪瘟的主要疫病之一。一些猪场伪狂犬病发病和流行比较严重,而且还有逐渐扩大的趋势,给养猪行业带来了巨大的挑战和经济损失。因此掌握伪狂犬病病毒的临床诊断和防控措施很有必要。本文则对猪伪狂犬病病毒的抗体检测进行了阐述,希望对猪伪狂犬病的诊断有所帮助。     

家兔人工感染疑似猪伪狂犬病病科的病理学观察

采用石河子地区某猪场疑似伪狂犬病病猪脑悬 液人工接种于家兔,从被感染家兔表现不可忍受的奇痒,发热典型症状,剖检和病理组织学及包涵体的观察结果表明,该猪场的患猪可诊断为伪狂犬病。     

猪伪狂犬病的防制体会

近几年随着养猪业的迅猛发展，猪的各种疾病也此起彼伏，猪伪狂犬病就是其中的一种，其感染率和发病率有扩大蔓延之势。     

猪伪狂犬病的诊断和控制

对广东某集约化猪场1993年10月流行的以母猪流产死胎和2周龄内仔猪以神经症状为主要表现的疫病进行诊断,经流行病学调查、临床诊断、动物试验和中和试验等方法确诊为伪狂犬病,应用哈尔滨兽医研究所生产的猪伪狂犬病弱毒疫苗作紧急免疫接种,2周内控制疫情.     

PCR检测伪狂犬病病毒的研究

用ＰＣＲ的方法，扩增了伪狂犬病病毒ｇｐ５０基因的一段较为保守区。检测了不同ＰＲＶ毒株感染的ＢＨＫ细胞以及接种的家兔，并对ＰＣＲ检测的灵敏性作了初步研究。结果表明，ＰＣＲ法扩增ｇｐ５０基因具有较高特异怀和灵敏性，是检测ＰＲＶ的有效方法。     

具有年龄结构的伪狂犬病模型分析

根据伪狂犬病在猪的各年龄段传播特性的不同,建立了具有成年和幼年两个年龄结构的伪狂犬病模型,并分析了其动力学行为,寻求决定疾病绝灭与否的基本再生数.当基本再生数小于或等于1时,模型仅有唯一的无病平衡点,当基本再生数大于1时,模型还存在唯一的正平衡点,利用线性化方法和Liapunov函数方法,讨论了两个平衡点的全局渐近稳定性.     

PRV闽A株gE基因抗原编码区片段在昆虫细胞中的表达研究

伪狂犬病病毒(PRV)糖蛋白E是一种在伪狂犬病的根除计划中具有重要作用的糖蛋白.将伪狂犬病病毒闽A株gE基因抗原编码区片段克隆到杆状病毒Bac-to-Bac系统表达载体pFastBacHTa中,获得的重组表达载体pFastBacHTa-FS转化大肠杆菌DH10BAc感受态细胞后,得到的重组Bacmid DNA在脂质体介导下转染粉蚊夜蛾Hi5细胞,通过细胞内同源重组,获得了含目的片段的重组病毒.此重组病毒感染Hi5细胞后72 h,细胞总蛋白的SDS-PAGE与Western-Blot结果表明,gE基因抗原编码区片段在Hi5细胞中得到了正确表达,表达产物约35000,占细胞总蛋白的9.31%.溶解性分析显示该片段在Hi5细胞中为不可溶性表达.     

伪狂犬病毒致病机理的研究进展

伪狂犬病是一种危害多种野生动物和家畜的急性病毒性传染病,猪为病毒的原始宿主和传染源,一旦感染则成潜伏感染状态,终身带毒.本病属于典型的且极难预防的自然疫源性传染病之一,给畜牧业发展带来很大的损失.文章就伪狂犬病毒的主要特性、致病机理等进行综述,并对其免疫机理和防制的研究前景做一展望.     

浅谈猪呼吸道疾病的诊断与防治

(1)病毒：猪流行性感冒病毒、猪瘟病毒、伪狂犬病病毒、猪繁殖-呼吸综合症病毒、包涵体鼻炎病毒、猪巨大细胞病毒、猪断奶肺炎综合症病毒。     

国内狂犬病传播的数学模型及控制策略

通过对目前狂犬病流行现状的研究，在考虑中国实际情况的基础上，建立了新的数学模型．本模型不仅涉及到狂犬病在犬类中的传播，还考虑了暴露和患病犬以及携带病毒的“健康犬”使人感染狂犬病的现象．通过寻找“基本再生数”对比了捕杀、免疫、捕杀和免疫相结合的3种不同策略在控制狂犬病传播中的有效性．分析和模拟结果表明，3种控制狂犬病的方法中捕杀的效果最好，免疫的效果次之，捕杀和免疫相结合的效果最差．同时根据中国目前城市和农村发展不平衡的现状，提出了在城市以免疫为主，在农村采用捕杀和免疫相结合的控制狂犬病的措施，从而为中国目前控制狂犬病的流行提供了理论依据．     

CpG ODN对初生仔猪外周血CD4+和CD8+T淋巴细胞的影响

以猪伪狂犬病毒为模式病毒,研究了CpG ODN对仔猪外周血中CD4^＋,CD8^＋T淋巴细胞的影响．实验结果表明：疫苗组初生仔猪血液中CD4^＋／CD8^＋T淋巴细胞比例随年龄的增加逐渐降低,而CpG ODN能有效抑制其降低（P〈0．05）;CpG ODN的使用也可以阻止伪狂犬病毒导致的血液中CD4^＋T淋巴细胞比例的降低;联合免疫CpG ODN和猪伪狂犬病毒活疫苗,可以诱导初生仔猪血清中猪伪狂犬病毒特异性抗体迅速产生并达到较高的水平（P〈0．05）;这表明CpG ODN能显著增强动物的免疫应答能力．     

新疆分离株伪狂犬病病毒的致病机理研究

采用从本地发病猪采集的伪狂犬病病毒组织对家兔进行了人工感染,每天观察感染家兔临床表现。20h后每隔一段时间处死1只兔子,取其扁桃体、淋巴结、肝脏、肾脏、脾脏、脑等组织制成石蜡切片。用免疫酶组织化学染色法检测病毒在组织中分布情况。结果表明：病毒首先在扁桃体内出现,淋巴结次之;40h后病毒分布于全身各组织脏器中,尤其在淋巴结、心脏、肾脏、肝脏中病毒大量存在。     

Cloning and Characterization of UL34 Gene of Pseudorabies Virus HB Isolate

In this study, China isolate HB of pseudorabies virus(PRV) was confirmed and genotypically characterized by amplifying and sequencing of partial UL34, a conservative gene involved in the egress of nucleocapsids from the nucleus, for phylogenetic analysis. The open reading frame(orf) of UL34 of PRV HB isolate is composed of 786 nucleotides, which encoded 262 amino acids. In addition, a potential transmembrane domain(241-260 aa) and 11 potential phosphorylation sites were also found in the UL34 of PRV HB isolate. Multiple amino acids alignment indicated that UL34 proteins of PRV strains derived from different geographic origins were highly conservative, but some mutations were also found. Phylogenetic analysis based on UL34 protein indicated that PRV HB strain was evolutionarily distinct from other recent China strains sequenced so far, forming a single clade within the phylogeny. Moreover, PRV HB isolate had close evolutionary relationship with Bo HV-1 and Bo HV-5 within the Alphaherpesvirinae. Taken together, these results indicated that PRV strains were in the progress of evolution. This study has expanded the knowledge of genetic profiles of PRV strains.     

Thymidine kinase gene mutation leads to reduced virulence of pseudorabies virus

To explore correlation between the tk gene structure of pseudorabies virus (PRV) and its virulence, to study the effect of the gene mutation on PRV biological properties, and to investigate mechinism of reduced virulence, thymidine kinase (TK)-deficient mutant of pseudorabies virus strain Hubei (PRV HB) was isolated by selection for resistance to 5-bromodeoxyuridine. The tk genes of PRV HB and its TK^― mutant were cloned and sequenced. 1587 base pairs of the tk gene and flanking regions of wild-type (wt) virus were sequenced, which included an open reading frame (ORF) of 1098 bp encoding a protein of 366 amino acids. The ORF contained two 137-bp repeated sequences, which were connected by an adenosine. 1458 bp of the tk and flanking regions of TK^― mutant were sequenced. Analysis of the tk gene sequence of TK^― mutant indicated that one of 137 bp repeated sequence and the connecting adenosine in the tk gene of the wt virus was deleted and a repeated sequence of 8 nucleotides (GCGCGCC) was inserted. All other nucleotides of TK^―mutant were identical to that of wt virus. Deletion and insertion of the nucleotide sequence resulted in a frameshift and a premature chain termination, and the resultant TK protein was not active. Analysis of the amino acid sequence revealed that TK protein of PRV HB contained the conserved consensus sequence of herpesviral TKs and an additional conserved-DHR-motif. The results of this work also indicated that TK^― mutant was genetically stable. Compared to PRV HB, virulence of TK^― mutant was greatly decreased. Mice vaccinated with TK^― mutant were completely protected against a lethal challenge with virulent PRV (HB).     

Cloning and characterization of <Emphasis Type="Italic">UL</Emphasis>34 gene of pseudorabies virus HB isolate

In this study, China isolate HB of pseudorabies virus (PRV) was confirmed and genotypically characterized by amplifying and sequencing of partial UL34, a conservative gene involved in the egress of nucleocapsids from the nucleus, for phylogenetic analysis. The open reading frame (orf) of UL34 of PRV HB isolate is composed of 786 nucleotides, which encoded 262 amino acids. In addition, a potential transmembrane domain (241-260 aa) and 11 potential phosphorylation sites were also found in the UL34 of PRV HB isolate. Multiple amino acids alignment indicated that UL34 proteins of PRV strains derived from different geographic origins were highly conservative, but some mutations were also found. Phylogenetic analysis based on UL34 protein indicated that PRV HB strain was evolutionarily distinct from other recent China strains sequenced so far, forming a single clade within the phylogeny. Moreover, PRV HB isolate had close evolutionary relationship with BoHV-1 and BoHV-5 within the Alphaherpesvirinae. Taken together, these results indicated that PRV strains were in the progress of evolution. This study has expanded the knowledge of genetic profiles of PRV strains.     

小鼠巨细胞病毒荧光定量PCR检测方法的建立及应用

目的 建立小鼠巨细胞病毒(Mouse Cytomegalovirus,MCMV)荧光定量PCR方法并对其进行初步应用。方法 选取NCBI发表的MCMV Smith株DNA polymerase基因保守序列设计引物探针,建立MCMV的荧光定量PCR方法,对方法的特异性、敏感性、重复性及稳定性进行验证,并应用该方法检测掺入MCMV的小鼠血液样品及2018年度送检的409份小鼠血液样本。结果 建立的MCMV荧光定量PCR方法标准曲线Slope为-3. 418,R2值为0. 999,扩增效率为96. 137%,可定量检测到的MCMV最低含量为47 copies/μL。以大鼠巨细胞病毒,猴巨细胞病毒,人单纯疱疹病毒,伪狂犬病毒及猫疱疹病毒I型为模板均无扩增曲线,特异性良好。方法组内和组间变异系数分别为0. 39%～0. 68%和0. 48%～1. 01%,重复性和稳定性好。可检测到掺入小鼠血液样品中MCMV病毒的最大稀释度为1∶1000(100. 75 TCID50/0. 1 m L),409份小鼠血液样品经检测均为阴性。结论 建立的小鼠巨细胞病毒荧光定量PCR方法有很好的敏感性、特异性及稳定性,可有效地检测小鼠中MCMV,为实验小鼠MCMV的监测及相关标准的补充完善提供了技术参考。     

Specific inhibition of herpesvirus ribonucleotide reductase by synthetic peptides

Ribonucleotide reductase is an essential enzyme for DNA synthesis in all prokaryotic and eukaryotic cells; it catalyses the reductive conversion of ribonucleotides to deoxyribonucleotides. Several herpesviruses including herpes simplex virus type 1 (HSV-1), HSV-2, pseudorabies virus (PRV), equine herpesvirus type 1 (EHV-1) and Epstein-Barr virus (EBV) have been found to induce novel ribonucleotide reductase activities. There is evidence that the HSV-1 ribonucleotide reductase activity is virus-encoded and essential for virus replication. This makes herpesvirus ribonucleotide reductases potential targets for antiviral chemotherapy. The HSV-1-encoded enzyme consists of two subunits: V136, the large subunit of relative molecular mass (Mr) 136,000 (136K) (RR1), which has been shown to be essential for enzyme activity, and V38, the small subunit (RR2) which forms a complex with the large subunit and is also likely to be essential for enzyme activity. Two particular features of the enzyme make it an attractive antiviral target. First, there is evidence for a common, highly conserved herpesvirus ribonucleotide reductase and second, the interaction between the large and small subunits may itself be exploitable. Here we identify a synthetic peptide which specifically inhibits the activity of virus-induced enzyme. We deduce that the mechanism of inhibition involves interference with the normal interaction between the two types of subunit.     

《梁山伯与祝英台》与《罗密欧与朱丽叶》调式比较

《梁山伯与祝英台》与《罗密欧与朱丽叶》两部作品的内容都取材于民间故事,都属于标题性音乐作品,作品曲式结构基本相同。不同之处是对造成悲剧原因的展示有着明显区别,主、副部主题的表现和运用截然相反,音乐的主基调有较大差异,作品的整体风格迥异。     

道教与魏晋南北朝文人的功利欲望

欲望是作家创作的内驱力,道教对中国古代文人的欲望有什么影响,这是研究中国古代文学和文学理论不能不思考的问题。本文从道教的功利性,文人纷纷走近道教,文学观念中的道教因素等方面,对道教与魏晋南北朝文人的功利欲望作些专门探讨。这无疑会拓展人们研究的新视界。