不同途径感染猪血球凝集性脑脊髓炎病毒的比较病理学

用传代鼠血球凝集性脑脊髓炎病毒接种液，经不同途径感染12头5日龄仔猪作比较病理学研究。结果表明，经脑内注射、前后肢肌肉注射、口服、口服兼滴鼻等途径接种的仔猪全部感染发病，并在接种后36～98h死亡。发病猪出现脑、脊髓的小静脉和毛细管充血、出血、水肿，神经原水肿、急性肿胀、急性液化及中央染色质溶解，胶质细胞轻微增生等非化脓性脑脊髓炎特征。口服兼滴鼻接种猪脑脊髓出血、脑水肿最明显，神经原变质也最重。脑内接种猪脑膜的血管周围出现明显的免疫细胞反应。     

猪血凝性脑脊髓炎病毒抗体的调查

应用血凝和血凝抑制试验检测了从吉林省部分地区采集的猪血清中血凝性脑脊髓炎病毒（HEV）抗体。结果，212份样品中有94份呈现HEV抗体阳性反应，阳性率高达44．3％。被采集血清的猪未表现临床症状，说明该地区的猪群中存在HEV隐性感染。     

Neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells on mice with experimental autoimmune encephalomyelitis

AIM: To explore the neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells (BM-NSCs) on the mice with experimental autoimmune encephalomyelitis (EAE). METHODS: Female C57BL/6 mice (8~10 weeks old, n=32) were immunized with myelin oligodendrocyte glycoprotein 35-55 (MOG_35-55) to establish chronic EAE model. The mice were randomly divided into control (ddH₂O) group, fasudil group, BM-NSCs group, and fasudil+BM-NSCs group. The clinical score and body weight were recorded every other day. The expression of neurotrophic factors was determined by immunofluorescence staining. RESULTS: In comparison with ddH₂O group, fasudil combined with BM-NSCs delayed onset and ameliorated severity of EAE. The numbers of brain-derived neurotrophic factor, glial cell-derived neurotrophic factor, nerve growth factor, neurotrophin-3 and ciliary neurotrophic factor positive cells in fasudil group, BM-NSCs group and fasudil+BM-NSCs group were all increased in various extents. In particularly, the expression of these neurotrophic factors in fasudil+BM-NSCs group was significantly higher than that in the mice treated with fasudil or BM-NSCs alone (P<0.01). CONCLUSION: Fasudil combined with BM-NSCs promotes the expression of neurotrophic factors and improves microenvironment of central nervous system, thus playing a positive role in neural restoration and regeneration through a synergistic and superimposed effect.     

1株高致病性血凝性脑脊髓炎病毒的分离与鉴定

利用细胞培养方法从临床表现神经症状的死亡仔猪脑组织内分离获得1株病毒,同时对该病毒进行电镜负染观察、昆明种小鼠致病性试验、RT-PCR检测和部分基因测序分析。电镜观察可见细胞培养物中有大量的直径约115 nm的冠状病毒样粒子;接种病毒的小鼠均出现典型的神经症状,死亡率100%;血凝性脑脊髓炎病毒(HEV)特异性引物RT-PCR扩增结果阳性;其S基因序列与GenBank中登录的HEV-67 N相应基因序列的同源性高达99.6%。结果表明,分离的病毒为猪血凝性脑脊髓炎病毒,分离株暂命名为HEV-CC-2007。     

A study of experimental autoimmune encephalomyelitis in dogs as a disease model for canine necrotizing encephalitis

In the present study, the use of dogs with experimental autoimmune encephalomyelitis (EAE) as a disease model for necrotizing encephalitis (NE) was assessed. Twelve healthy dogs were included in this study. Canine forebrain tissues (8 g), including white and grey matter, were homogenized with 4 mL of phosphate-buffered saline for 5 min in an ice bath. The suspension was emulsified with the same volume of Freund''s complete adjuvant containing 1 mg/mL of killed Mycobacterium tuberculosis H37Ra. Under sedation, each dog was injected subcutaneously with canine brain homogenate at four sites: two in the inguinal and two in the axillary regions. A second injection (booster) was administered to all the dogs using the same procedure 7 days after the first injection. Clinical assessment, magnetic resonance imaging, cerebrospinal fluid analyses, necropsies, and histopathological and immunohistochemical examinations were performed for the dogs with EAE. Out of the 12 animals, seven (58%) developed clinically manifest EAE at various times after immunization. Characteristics of canine EAE models were very similar to canine NE, suggesting that canine EAE can be a disease model for NE in dogs.     

Effect of idazoxan on permeability of blood-brain barrier and expression of MMP-9/TIMP-1 in mouse experimental autoimmune encephalomyelitis

AIM：To study the effect of idazoxan (IDA) on the permeability of blood-brain barrier (BBB) and the expression of matrix metalloproteinase 9 (MMP-9) and tissue inhibitor of metalloproteinase 1 (TIMP-1) in mouse experimental autoimmune encephalomyelitis (EAE).METHODS：Female C57BL/6 mice (n=36) were randomly divided into control group, EAE group and IDA group, with 12 mice in each group. EAE was induced by myelin oligodendrocyte glycoprotein 35-55 (MOG35-55). IDA (2 mg/kg, ip, bid) was administered for 15 d after immunization. The neurological defects of the mice were observed daily and scored. The pathological changes were observed under microscope with HE staining and LFB myelin staining. The BBB permeability was detected by Evans blue extravasation. The expression of MMP-9 and TIMP-1 in the brain of EAE mice was determined by Western blotting.RESULTS：Compared with EAE group, the score of neurological defects in IDA group was decreased, the inflammation was relieved, the BBB permeability was reduced, and the expression MMP-9 and the ratio of MMP-9/TIMP-1 were decreased (P<0.05).CONCLUSION：The neuroprotective effect of IDA on mouse EAE might be related to the down-regulation of MMP-9 and the ratio of MMP-9/TIMP-1, thus reducing the degradation of BBB and the permeability of BBB, and ameliorating the pathologic process of EAE.     

Human Exposure to Rabid Free‐Ranging Cats: A Continuing Public Health Concern in Pennsylvania

Rabid free‐ranging cats have been a public health concern in Pennsylvania since raccoon variant rabies first was recognized in the state in the early 1980s. Over the last decade, between 1.5 and 2.5% of cats submitted to Pennsylvania's state laboratories for rabies testing have been positive. In this report, we describe the extent of rabies in free‐ranging cats in Pennsylvania. We also present two examples of human exposure to rabid free‐ranging cats that occurred in Pennsylvania during 2010–2011 and the public health actions taken to address rabies exposure in the humans and animals. We then describe the concerns surrounding the unvaccinated and free‐ranging cat population in Pennsylvania and possible options in managing this public and animal health problem.     

免疫禽脑脊髓炎弱毒疫苗后肉种鸡琼扩抗体的动态变化

为探讨肉种鸡免疫禽脑脊髓炎(AE)弱毒疫苗的抗体变化规律及其与后裔雏鸡对禽脑脊髓炎病毒(AEV)易感性的相互关系，本研究用琼脂扩散试验对14周龄免疫禽脑脊髓炎(AE)弱毒疫苗的5群艾维茵父母代肉种鸡，其中1群于34周龄加免AE油乳剂灭活苗，从20至60周龄对其AE抗体进行动态监测。结果发现：1)单免弱毒疫苗的4群种鸡，其AE抗体阳性率呈双低谷曲线变化规律，32周龄之前较高，为95％～83.4％；36～40周龄降至67.5％～71．1％，最低可至56．7％；44周龄转而升至80％以上，48～52周龄再度降至59．1％～67．5％：56周龄以后又呈上升(84．2％)。2)AE抗体低谷期间种鸡产蛋率明显降低，其后裔雏鸡AE母源抗体明显降低，而且衰减快。14日龄颈部皮下注射AEV Van Roekel，50％的雏鸡在攻毒后13d发生AE。有些雏鸡在14～21日龄自然发生AE。3)加免AE油乳剂灭活苗可避免种鸡AE抗体低谷期的出现。以上结果表明，在我国养鸡实际生产中种鸡开产前免疫1次AE弱毒疫苗，不足以为整个产蛋期提供有效保护，AE弱毒疫苗和灭活油苗联合应用是预防AE的有效措施。     

基因芯片技术检测5种马病毒

通过分子克隆技术获得马疱疹病毒1型(Equineherpesvirus1,EHV1)、马动脉炎病毒(Equinearteritisvirus,EAV)、马流感病毒(Equineinfluenzavirus,EIV)、马传染性贫血病毒(Equineinfectiousanaemiavirus,EIAV)和东部马脑脊髓炎病毒(East-ernequineencephalomyelitisvirus,EEEV)等5种病毒各一段高度保守的特异性基因片段,用芯片点样仪逐点分配到处理过的玻片上,制备成检测芯片。提取样品中的RNA,进行反转录和荧光标记后滴加到芯片上进行特异性杂交,对杂交结果进行扫描检测和计算机软件分析。结果显示,制备的基因芯片可同时检测和鉴别上述5种病毒,可检测到阳性杂交信号的最高稀释度为10-6的病毒液,约25个病毒DNA拷贝,但其它病毒材料未见红色荧光信号,证明了本方法的特异性。在进口马的隔离检疫期间,采集马鼻肺炎、马动脉炎中和抗体阳性但病毒分离阴性马匹的白细胞悬液,分别在EHV1和EAV位点处可检测到阳性杂交信号。证明基因芯片技术不但快速、准确和敏感,而且可同时进行多种病毒的检测。     

Host immune responses after administration of inactivated Venezuelan equine encephalomyelitis virus vaccines—III. Kinetics for neutralizing antibody immunoglobulin class responses in donors and adoptively-immunized recipients

C57B16 mice were immunized with either live, attenuated TC-83 strain VEE virus vaccine or formalin-inactivated VEE vaccine combined with Bordetella pertussis. The kinetics of specific donor and adoptively-immunized recipient anti-VEE neutralizing antibody responses were studied. Donor mice immunized with either live or inactivated VEE virus vaccine combined with potent adjuvants develop specific anti-VEE IgM and IgG responses as early as 7 days post-immunization. Anti-VEE IgM antibody responses comprise the majority of anti-VEE neutralizing antibody at this early time period. By 14 to 21 days post-immunization, anti-VEE IgG responses predominated. When adoptively-immunized recipients were studied, the anti-VEE IgM to IgG predominance seen in donors early after administration was reversed, and for each time-period studied, recipients' serum anti-VEE antibody class responses consisted principally of IgG rather than IgM antibody. Since T-cells cooperation with B-cells is critical in the IgM-IgG antibody shift, these studies support the critical role T-cells exert in adoptive transfer in a murine model of experimental VEE infection. Furthermore, immunization with either live or inactivated VEE vaccine coupled to a potent adjuvant induce comparable donor and adoptively-immunized recipient anti-VEE antibody class responses.