绵羊进行性肺炎病毒与山羊关节炎一脑炎病毒的血清学交叉反应的研究

本实验用琼脂凝胶免疫扩散试验（ＡＧＩＤＴ）和免疫印迹试验（ＩＢＡ）对实验感染绵羊进行性肺炎病毒（ＯＰＰＶ）的山羊血清与山羊关节炎—脑炎病毒（ＣＡＥＶ）抗原以及实验感染ＣＡＥＶ的绵羊血清与ＯＰＰＶ抗原的交叉反应进行了研究。４只接种ＯＰＰＶ的山羊中有一只山羊的血清可与ＣＡＥＶ琼扩抗原发生交叉反应,并在免疫印迹试验中可识别ＣＡＥＶ的ｇｐ４４、ｐ３５和ｐ２８。２只接种ＣＡＥＶ的绵羊中有一只绵羊的血清可与ＯＰＰＶ琼扩抗原发生交叉反应,并在免疫印迹试验中可识别ＯＰＰＶ的ｇｐ４４和ｐ２８。以上的交叉反应结果表明ＯＰＰＶ与ＣＡＥＶ的抗原之间具有密切的相关性,这对于ＯＰＰＶ通过山羊和ＣＡＥＶ通过绵羊的传代研究是非常重要的,并对将来的免疫预防策略具有重要的指导意义。     

山羊关节炎—脑炎与其相关病的血清学关系

用琼脂扩散法检测，山羊关节炎－脑炎病毒（ＣＡＥＶ）抗原与马传染性贫血（ＥＩＡ）阳性血清不发生反应，ＥＩＡＶ抗原与ＣＡＥ阳性血清亦不发生反应，但ＯＰＰＶ抗原与ＣＡＥ阳性血清发生反应。     

山羊关节炎－脑炎与其相关病的血清学关系

用琼脂扩散法检测，山羊关节炎－脑炎病毒（ＣＡＥＶ）抗原与马传染性贫血（ＥＩＡ）阳性血清不发生反应，ＥＩＡＶ抗原与ＣＡＥ阳性血清亦不发生反应．ＣＡＥＶ抗原与绵羊进行性肺炎（ＯＰＰ）阳性血清不发生反应．但ＯＰＰＶ抗原与ＣＡＥ阳性血清发生反应．     

山羊关节炎脑炎间接ELISA诊断方法的建立

山羊关节炎脑炎间接ＥＬＩＳＡ诊断方法的建立赵亚光王振漪沈荣显（中国农科院生物技术研究中心）（中国农科院研究生院）（中国农科院哈尔滨兽医研究所）山羊关节炎脑炎（ＣＡＥ）是８０年代新发现的一种由反转录病毒科慢病毒亚科成员ＣＡＥ病毒引起的山羊慢病毒传染病。...     

山羊关节炎-脑炎病毒对山羊人工感染试验

山羊关节炎－脑炎病毒对山羊人工感染试验王治才龚成润李建军马文戈胡泽渊（新疆畜牧科学院兽医所）山羊关节炎－脑炎（ＣＡＥ）是由反录病毒科慢病毒亚科的ＣＡＥ病毒引起的奶山羊的一种慢性传染病。该病在世界各地广为流行。欧美一些国家奶山羊ＣＡＥ阳性率达６５～８０...     

实验感染山羊关节炎—脑炎病毒的绵羊抗体应答反应的研究

本实验用琼脂凝胶免疫扩散试验和免疫印迹试验对实验感染山羊关节炎—脑炎病毒（ＣＡＥＶ）的绵羊抗体应答反应进行了研究，用两种方法都可在接毒绵羊的血清中检测到ＣＡＥＶ的抗体。琼脂凝胶免疫扩散试验最早可于接毒后的第７周时检测到抗体，免疫印迹试验最早可于接毒后的第６周时检测到抗ＣＡＥＶ的ｇｐ１２５、ｇｐ４４、ｐ３５、ｐ２８和ｐ１４的抗体，这说明免疫印迹试验更为敏感一些。本实验的结果表明ＣＡＥＶ可在绵羊体内诱生明显的体液免疫应答反应，因此用ＣＡＥＶ通过绵羊体传代的方法可能会得到具有良好的抗原性的ＣＡＥＶ毒株，这对于人工培养ＣＡＥＶ强毒是非常重要的。此外，本实验还为ＣＡＥＶ通过绵羊体传代的研究提供了非常实用的检测手段     

山羊胎肺细胞增殖山羊关节炎：脑炎病毒制备琼扩抗原的试验

试图用山羊胎肺细胞增殖山羊关节炎－脑炎病毒获得成功，将含毒培养液用ＰＥＧ沉淀制成琼扩抗原，经与羔羊滑膜细胞增残ＣＡＥＶ制备的抗原比较，主要蛋白成分和性能一致，具有特异性强，稳定性好等特点。结果表明，用山羊胎肺细胞增殖ＣＡＥＶ制备琼扩抗原是一种可供选择的有效方法。     

绵羊进行性肺炎病毒与山羊关节炎一脑炎病毒的血清学交以应的研究

本实验用琼脂凝胶免疫扩散试验和免疫印迹试验对实验感染绵羊进行肺性炎病毒的山羊血清与山羊关节炎－脑炎病毒抗原以及实验感染ＣＡＥＶ的棉羊血清与ＯＰＰＶ抗原的交叉反应进行了研究。     

一日龄雏鸡感染鸡盆血病病毒及鸡呼肠病毒的协同作用

１日龄ＳＰＦ来航鸡经口双重感染鸡贫血病病毒（ＣＡＶ）Ｃｕｘ－１株及鸡呼肠病毒（ＲＥＯＶ）Ｓ１１３３或Ｕｃｈｉｄａ株。于接种后第１４天，检查血细胞压积（ＰＣＶ）、称体重、观察骨髓、胸腺及法氏囊病变。结果，双重感染ＣＡＶ及Ｓ１１３３株ＲＥＯＶ的雏鸡与感染其中之一病毒的雏鸡相比，其体重增重少，病变重，而且比仅感染ＣＡＶ的雏鸡在平均ＰＣＶ上也明显低。双重感染ＣＡＶ及Ｕｃｈｉｄａ株ＲＥＯＶ的雏鸡，不加重发病     

ELISAA法检测犬腹泻粪样中的犬冠状病毒

用ＦＥ细胞增殖犬冠状病毒（ＣＣＶ）参考株，分别免疫家兔和ＢＡＬＢ／ｃ小鼠制备ＣＣＶ多抗和单抗，建立了夹心ＥＬＩＳＡ及Ｄｏｔ－ＥＬＩＳＡ诊断方法。在检测的８４例犬腹泻粪样中，多抗、单抗夹心法显示ＣＣＶ阳性１６例，Ｄｏｔ－ＥＬＩＳＡ阳性１３便，后１３例包括在前１６例中，从８４例腹泻犬粪样中随机取３例作ＣＣＶ、犬细小病毒（ＣＰＶ）双项检测，ＣＣＶ阳性１６例，ＣＰＶ阳性６例，ＣＣＶ、ＣＰＶ混合感染４例。结     

Immunomodulation of caprine lentiviral infection by interleukin-16

Interleukin-16 (IL-16) is a proinflammatory cytokine produced by a variety of cells including lymphocytes, macrophages, mast cells, and eosinophils. We have shown in our previous studies increased expression of IL-16 mRNA and protein in caprine arthritis-encephalitis virus (CAEV)-infected goats blood. In this study, we determined the immunomodulatory effects of IL-16 in vitro using cells derived from CAEV infected and uninfected goats. Human recombinant IL-16 (rhIL-16) significantly increased chemotaxis of peripheral blood mononuclear cells (PBMCs) of both control and CAEV-infected goats. Pretreatment of PBMC with anti-goat CD4 monoclonal antibody inhibited IL-16-induced chemotaxis of PBMC of control and infected goats suggesting that IL-16 exerts its action in goats primarily by binding to CD4. The CAEV proviral DNA was less in caprine monocytes treated with rhIL-16 infected in vitro with CAEV. These data suggest inhibitory effect of IL-16 on viral integration. Flow cytometric studies indicated a trend toward IL-16-induced increased expression of lymphocyte activation markers. Combined with our previously reported data, these experiments suggest that increased IL-16 expression during CAEV infection may inhibit viral integration.     

Adult neurogenesis in the mammalian dentate gyrus

Earlier observations in neuroscience suggested that no new neurons form in the mature central nervous system. Evidence now indicates that new neurons do form in the adult mammalian brain. Two regions of the mature mammalian brain generate new neurons: (a) the border of the lateral ventricles of the brain (subventricular zone) and (b) the subgranular zone (SGZ) of the dentate gyrus of the hippocampus. This review focuses only on new neuron formation in the dentate gyrus of the hippocampus. During normal prenatal and early postnatal development, neural stem cells (NSCs) give rise to differentiated neurons. NSCs persist in the dentate gyrus SGZ, undergoing cell division, with some daughter cells differentiating into functional neurons that participate in learning and memory and general cognition through integration into pre-existing neural networks. Axons, which emanate from neurons in the entorhinal cortex, synapse with dendrites of the granule cells (small neurons) of the dentate gyrus. Axons from granule cells synapse with pyramidal cells in the hippocampal CA3 region, which send axons to synapse with CA1 hippocampal pyramidal cells that send their axons out of the hippocampus proper. Adult neurogenesis includes proliferation, differentiation, migration, the death of some newly formed cells and final integration of surviving cells into neural networks. We summarise these processes in adult mammalian hippocampal neurogenesis and discuss the roles of major signalling molecules that influence neurogenesis, including neurotransmitters and some hormones. The recent controversy raised concerning whether or not adult neurogenesis occurs in humans also is discussed.     

pGh9:ISS1 transpositional mutations in Streptococcus uberis UT888 causes reduced bacterial adherence to and internalization into bovine mammary epithelial cells

Streptococcus uberis is an important mastitis pathogen that affects dairy cows worldwide. In spite of the economic impact caused by the high prevalence of S. uberis intramammary infections (IMI) in many well-managed dairy herds, pathogenic strategies and associated virulence factors of S. uberis are not well understood. It has been shown that S. uberis attaches to and internalizes into mammary epithelial cells and can survive inside cells for extended periods of time. We hypothesize that early attachment to and internalization into mammary epithelial cells is a critical step for the establishment of intramammary infection. The aim of this study is to identify and characterize chromosomally encoded virulence factors of S. uberis that allow early bacterial attachment to and internalization into mammary epithelial cells. A common approach used to identify virulence factors is by generating random insertion mutants that are defective in adherence to and internalization into mammary epithelial cells using pGh9:ISS1 mutagenesis system. A random insertion mutant library of S. uberis strain UT888 was created using a thermo-sensitive plasmid pGh9:ISS1 carrying ISS1 insertion sequence. Integration of the insertion sequence into the chromosome of these mutant clones was confirmed by PCR and Southern blot. Southern blot analysis of mutant clones also showed that insertional integration was random. Of 1000 random chromosomal insertion mutants of S. uberis strain UT888 screened, 32 had significantly reduced ability to adhere to and internalize into mammary epithelial cells. Chromosomal mapping of insertion sequence integration sites in some of these defective mutants showed integration into penicillin binding protein 2A (pbp2A), sensor histidine kinase, tetR family regulatory protein, phosphoribosylaminoimidazole carboxylase catalytic subunit (purE), lactose phosphotransferase, phosphoribosylamine glycine ligase (purD), and other genes involved in metabolic activities. These proteins may have a significant role in early bacterial colonization of the mammary gland during infection.     

猪瘟病毒E0和E2蛋白融合表达的重组腺病毒构建及免疫保护性研究

为评价重组腺病毒融合表达猪瘟病毒(CSFV)E0和E2蛋白的免疫保护效果,本研究以CSFV基因组为模板,应用RT-PCR扩增E0和E2蛋白的编码基因,通过pET-32a载体将E0和E2基因串联,形成pET-E0-E2重组质粒。用KpnⅠ和NotⅠ双酶切pET-E0-E2得到E0-E2融合基因,定向亚克隆于穿梭载体pAdTrack-CMV中,采用"两步转化法"在细菌内同源重组,构建携带E0-E2基因的重组腺病毒转移载体质粒pAdEasy-E0-E2,经PacⅠ酶切线性化后转染人胚胎肾细胞(HEK293),成功包装出重组腺病毒(rAd-E0-E2),PCR和western blot检测表明,E0-E2基因已重组于腺病毒基因组中并获得表达。将rAd-E0-E2接种于小鼠和猪,并通过ELISA进行抗体检测。另外,将rAd-E0-E2经肌肉2次(间隔7d)免疫接种6周龄～7周龄猪,3周后用103TCID50CSFV石门株攻毒。结果表明,rAd-E0-E2免疫组6/7头存活,各组织器官带毒时间不超过10d,而非重组腺病毒rAd-CMV免疫组和空白对照组全部死亡,各组织器官均能分离到CSFV。结果提示,rAd-E0-E2能使免疫猪抵抗CSFV强毒攻击,为进一步研制猪瘟基因工程疫苗提供了实验依据。     

重组逆转录病毒介导的neo~R基因在公鼠生殖系细胞的转移

将感染滴度为 10 6cfu/ m L、携带新霉素磷酸转移酶 (neo R)基因的重组逆转录病毒与台盼蓝、polybrene按一定比例配制成注射液 ,单侧睾丸注入 14～ 16日龄公鼠曲细精管以体内感染精原干细胞 ,进行重组逆转录病毒 -精原干细胞介导的基因转移研究。注射后 45 d,采集公鼠精液 ,进行精液品质与 PCR检测。结果表明 ,在优化的注射条件下 ,曲细精管内注射重组逆转录病毒后 ,不会干扰小鼠精子的正常发育能力 ,并能够将外源基因整合到精子基因组 ,从而首次通过重组逆转录病毒成功地实现了 neo R基因在公鼠生殖系细胞的转移     

精子载体法获得转人her2基因猪

将带有外源基因的载体酶切纯化后与去精清的猪新鲜精液按比例混合，17℃静置处理使外源DNA进入精子头部，通过人工授精使受体母猪怀孕后产下20头仔猪，经PCR特异性片段扩增，特异片段序列测定和比对，共4头PCR结果为阳性。初步证明人her2基因已在猪染色体上整合，其整合率约为20％。本研究利用精子载体法成功获得了转基因阳性猪，为大动物的转基因研究提供了理论与实践的参考。     

家蚕粗线期染色体核型模式的建立

家蚕染色体核型分析是研究家蚕遗传、变异和基因定位等的重要方法之一。用Luc ia染色体分析软件对家蚕品种大造卵母细胞粗线期染色体的长度和染色粒分布情况作了调查,结果表明:不同细胞的染色体总长度差异较大,同一细胞内各染色体绝对长度也互不相同,而且不同细胞相同序号染色体之间的长度差异也很大,但不同细胞相同序号染色体的相对长度比较接近,差异较小,适合作为建立核型模式的参数。对家蚕粗线期染色体的染色粒分析表明:染色粒在整个染色体群的各条染色体中含量均较高,染色粒相对长度在1%~3%之间的染色体比例最大。综合4种参数,建立了一个家蚕粗线期染色体的核型模式图,初步反映粗线期每条染色体各自的特征。     

牧场整合下的牧户博弈结构分析与牧场治理反思——以法治与“公地”治理理论结合为视角

面对当前牧场家庭承包经营困境,得到广泛支持的治理策略是牧场整合。得到经验、政策和理论研究支持的牧场整合方式有家庭牧场、联合牧场、合作牧场和股份牧场。本文以法治和"公地"治理理论结合的角度深入分析了各种整合方式,认为家庭牧场、联合牧场没有超越家庭承包经营的范围,并非理想的牧场整合方式,股份牧场适用牧场整合的范围很有限,只有牧户自主组成的合作社为整合主体的牧场整合才是适宜当前法律框架的牧场治理模式,但家庭牧场、联合牧场仍是不可或缺的过渡替代方案。     

碱胁迫对羊草克隆整合的影响

采用盆栽试验研究了碱对克隆植物羊草(Leymus chinensis)生长的影响。将子株分别置于4个水平梯度的碱胁迫处理下,母株保持无任何碱胁迫处理,并对子株与母株之间的根茎进行连接和切断处理。结果发现,克隆整合明显增加了高碱处理子株的存活数、生物量、分蘖数、高度和叶水势,但低碱处理影响并不明显。克隆整合使地下组织器官根茎芽数量明显增加,根重有所下降,根茎长没有显著变化。受到碱胁迫的子株虽然明显受益克隆整合,但母株也有一定的损耗。克隆整合是克隆植物适应非均一碱胁迫的重要适应对策。     

Origin of the neuro‐sensory system: new and expected insights from sponges

The capacity of all cells to respond to stimuli implies the conduction of information at least over short distances. In multicellular organisms, more complex systems of integration and coordination of activities are necessary. In most animals, the processing of information is performed by a nervous system. Among the most basal taxa, sponges are nerveless so that it is traditionally assumed that the integrated neuro‐sensory system originated only once in Eumetazoa, a hypothesis not in agreement with some recent phylogenomic studies. The aim of this review is to show that recent data on sponges might provide clues for understanding the origin of this complex system. First, sponges are able to react to external stimuli, and some of them display spontaneous movement activities. These coordinated behaviors involve nervous system‐like mechanisms, such as action potentials and/or neurotransmitters. Second, genomic analyses show that sponges possess genes orthologous to those involved in the patterning or functioning of the neuro‐sensory system in Eumetazoa. Finally, some of these genes are expressed in specific cells (flask cells, choanocytes). Together with ultrastructural data, this gives rise to challenging hypotheses concerning cell types that might play neuro‐sensory‐like roles in sponges.