Experimental rabies in skunks: oral, nasal, tracheal and intestinal exposure.

Striped skunks (Mephitis mephitis) were exposed to challenge virus standard rabies virus by feeding infected mouse brain in suspension or as intact brain free choice, by forced feeding of suspension, and by intranasal, intratracheal and intraintestinal instillation of suspension. All of five skunks exposed intranasally, two of five exposed intratracheally and two of ten exposed by forced feeding developed rabies. None of the skunks exposed to challenge virus standard virus, by other methods, became rabid. Most of the survivors, when challenged intramuscularly with street rabies virus at six months, developed rabies. The results indicate that the skunk is much more susceptible to challenge virus standard rabies virus given intranasally than by the other methods used. When disease occurs following oral administration, infection may be associated with prolonged contact with buccal mucosa or accidental contact with nasal mucosa. Survivors had little or no protection when challenged intramuscularly with street rabies virus.     

Comparative Analysis of the Pathogenic Mechanisms of Street Rabies Virus Strains with Different Virulence Levels

Objective To characterize two strains of street rabies virus （RABV） isolated from the brain tissue of cattle from Inner Mongolia. Differences in the histopathological and ultrastructural changes in the brain tissue of infected mice were determired to reveal variation in the pathogenesis of infection between street rabies virus strains. Methods Ten-day-old mice were intracranially inoculated with one of three virus strains and brain tissue harvested when the mice were moribund. Various histopathological and ultrastructural markers of disease were then compared between the groups. Results Infection with the street virus strain CNMllO1C resulted in severe neuronal dendrites damage, but only mild cell apoptosis, T lymphocyte infiltration and microglial activation. Infection with the other street virus strain, CNMll03C, was characterized by cell apoptosis, T lymphocyte infiltration and microglial activation as well as dendrites damage. However, in comparison, infection with the attenuated virus strain CTN caused ~evere T lymphocyte infiltration, microglial activation and cell apoptosis, but left the neuronal dendrites intact. Conclusion The two street rabies virJs strains isolated from cattle from Inner Mongolia had different levels of virulence and caused distinct pathological changes in infected mice. Therefore, we concluded that different pathogenic mechanisms exist between different RABV strains.     

风疹病毒在BALB/c小鼠体内的感染与分布

用BALB/c小鼠形成风疹病毒感染模型,小鼠经尾静脉感染RVGOS_(-10)株10~6TCID_(50),每只0.5ml,病毒血症持续约一周,感染后第4天达最高滴度,为10~(6.3)TCID_(50)/0.25ml,间接免疫荧光及间接酶标染色法观察小鼠组织切片,感染后不同时间,病毒对小鼠各脏器可形成广泛的侵害,抗原主要分布于胞桨,病毒分离结果与之吻合,感染后第18天,心、眼、肺中的病毒滴度分别为10~(5.2),10~(5.6),10~(5.5)TCID_(50)。     

狂犬病病毒街毒株对体外培养小鼠海马神经元的损伤作用及其机制

目的：探讨狂犬病病毒街毒株（RV）感染神经细胞的形态学表现,揭示狂犬病病毒致神经功能异常的机制。方法：体内实验,20只C57/BL小鼠随机分为实验组和对照组,每组10只,实验组小鼠脑内接种30 μL　RV病毒液（10TCID50）,对照组小鼠脑内接种等量细胞维持液,利用抗RV抗体检测病毒抗原在小鼠脑组织的分布。体外实验,培养原代海马神经细胞,培养1周后,感染狂犬病病毒,免疫荧光检测感染72、96和120 h后病毒的增殖情况。结果：体内实验,狂犬病病毒感染4 d后,在海马CA1区锥状神经元胞体和树突中均能检测到狂犬病毒抗原;狂犬病病毒感染7 d后,仅在海马CA1区胞体中检测到狂犬病病毒抗原。体外实验,狂犬病RV感染体外培养的原代神经细胞120 h后,感染的神经元数量最多,且感染的神经元树突数量减少。结论：狂犬病RV可感染、损伤海马CA1区树突,从而引起小鼠神经功能异常。     

Experimental rabies in skunks: effects of immunosuppression induced by cyclophosphamide

Striped skunks (Mephitis mephitis) were inoculated with street rabies virus and immunosuppressed with several doses of cyclophosphamide. Control skunks were inoculated with street virus only. The skunks were killed in terminal stages of the disease and several tissues were collected for examination by immunofluorescence, light microscopy and viral titration. Sera collected at euthanasia from most of the principals did not contain detectable rabies neutralizing antibodies, whereas high titers occurred terminally in controls. Immunofluorescence was much more entensive in submandibular salivary glands of cyclophosphamide-treated than control skunks. Similarly, virus was isolated from this tissue more consistently and at higher titer from principals than from controls. Immunofluorescence was extensive in brains of all skunks (both groups), but virus was isolated consistently only from brains of cyclophosphamide-treated skunks. Most of the cyclophosphamide-treated skunks had very few inflammatory cells in brain and cerebrospinal ganglia. Neuronal degeneration occurred in dorsal root ganglia of both principals and controls. The results suggest that the immune response has no effect on the development of rabies-induced aggressive behavior, that the immune response may inhibit salivary gland infection and that it is not essential for the development of neuronal degeneration in dorsal root ganglia.     

口康含漱液治疗急性白血病并口腔感染的临床观察及体外抑菌实验研究

目的：研究口康含漱液的体外抑菌作用及临床疗效。方法：用琼脂稀释法测定口康含漱液对49株口腔病菌的最低抑菌浓度，并对60例急性白血病并口腔感染患者中40例采用中药口康含漱液治疗，20例采用口泰含漱液治疗。结果：49株口腔病菌中，27株革兰氏阴性杆菌MIC范围0．03－1．0g/ml,MIC50均为0．125g/ml,MIC90为0．125－0．50g/ml；21株革兰氏阳性球菌MIC范围0．008－0．50g/ml,MIC50 09.015-0.125g/ml,MIC90为0．015－0．50g/ml,1株白色念珠菌MIC 0．015g/ml.40例急性白血病并口腔感染患者采用中药口康含漱液治愈率为82．5％，总有效率92．5％，较口泰对照组明显升高（P＜0．01），结论：口康含漱液对各类不同口腔病菌株均有抑菌作用，临床应用对急性白血病并口腔感染患者疗效明显优于口泰对照组，是一种较好的抗口腔感染的药物。     

用浓缩地鼠肾细胞狂犬病毒抗原免疫马匹获得高效价血浆

目的制备高效价马抗狂犬病免疫血浆。方法通过细胞制备、病毒的接种、培养、收获、浓缩等步骤，研制了浓缩地鼠。肾细胞狂犬病毒抗原，并与羊脑狂犬病毒抗原作马匹免疫对比试验。结果浓缩地鼠。肾细胞抗狂犬病免疫血浆的效价明显高于羊脑抗狂犬病免疫血浆。试验组血浆平均效价为157IU／ml，对照组为113IU／ml，两组差异有统计学意义。结论已成功制备高效价抗狂犬病免疫血浆。     

流行性出血热病毒感染黑线姬鼠有效剂量的观察

用Hantaan病毒(76-118株)悬液以不同感染剂量采用灌胃和浸泡毒米任实验鼠自行取食两种方法,对捕自非疫区的黑线姬鼠进行实验感染。结果表明:实验鼠未能通过完整的消化道粘膜感染,但可通过破损的消化道粘膜感染,感染剂量与感染率呈正相关,最低感染剂量为10ID_(so)/ml。     

西咪替丁、维生素C小鼠体内抗狂犬病效果研究

目的 旨在寻找一种安全有效的抗狂犬病毒药物,为狂犬病的治疗提供参考.方法 采用小鼠动物模型的随机对照实验,实验组腹腔注射0.4 mg/只、3.2 mg/只西咪替丁及50 mg/只维生素C,用生存分析比较实验组和对照组生存差异,评价药物的疗效.结论 0.4 mg/只和3.2 mg/只的西咪替丁药量治疗狂犬病毒感染的小鼠,实验组和对照组生存率均无显著统计学差异(P>0.05);维生素C 50 mg/只剂量经腹腔给药,其实验组与对照组生存率也无显著的统计学差异.结论在小鼠狂犬病模型中并没有发现西咪替丁及维生素C具有抗狂犬病毒的作用,目前仍然没有单一有效的抗狂犬病毒药物.     

人季节性H1N1流感病毒小鼠感染模型的建立

目的制备人季节性H1N1流感病毒的小鼠感染模型,为研究流感病毒致病性、研发抗病毒药物提供模型动物。方法将人季节性H1N1流感病毒在鸡胚尿囊腔扩增后,滴鼻接种小鼠,4 d后将小鼠处死,挑选感染体征严重者进行实时荧光PCR(FQ-PCR)检测肺中的流感病毒,将检测阳性的肺上清在鸡胚尿囊腔扩增,接种于下一代小鼠。比较各代小鼠对流感病毒的适应情况,直至小鼠出现明显的感染体征,取肺研磨制成匀浆,获得流感病毒鼠肺适应株并检测其半数致死量(LD50)。将10 LD50的病毒液接种于小鼠,建立人季节性H1N1流感病毒的小鼠感染模型,观察模型小鼠的一般活动状态、体质量变化、肺部病变,HE染色观察肺部病理切片,计算肺指数,FQ-PCR检测病毒RNA。结果人季节性流感病毒在小鼠体内传代4次后,鼠肺适应株制备完成,其经鼻的LD50为10-2.41/0.05 mL。人季节性流感病毒的小鼠感染模型,一般状态差,体质量明显减轻,肺指数增大,70%出现死亡。病理切片观察病变明显,FQ-PCR显示流感病毒阳性。结论成功建立了人季节性H1N1流感病毒的小鼠感染模型。     

pH影响丁卡因麻醉作用实验研究

不同pH丁卡因溶液抑制离体坐骨神经动作电位和对家兔鼻粘膜、角膜的麻醉作用。pH7.4和4.8丁卡因溶液抑制离体坐骨神经动作电位IC50分别为0.34mg/ml和0.67mg/ml。对鼻粘膜麻醉作用ED50分别为0.39±0.03mg和1.1±0.15mg。对角膜麻醉作用ED50分别为0.45±0.05mg和1.2±0.18mg。上述三种实验均有非常显著差别。无论pH7.4还是4.8丁卡因溶液鼻粘膜给药1/5LD50后60min内血压、呼吸和心率均无显著影响     

狂犬病毒非临床型感染初探

目的：探讨狂犬病毒非临床型感染的可能性。方法：收集病例资料和检测暴露人群血清中的狂犬病毒抗体。结果：(1)收集到的2例患者，在被犬咬伤，愈后又复发不良反应，同时伴有血清中狂犬病毒抗体4倍增长的现象；(2)检测的暴露人群49例中(无疫苗接种史)，检出狂犬病毒抗体7例(14．28％)。结论：狂犬病毒可以引起非临床型感染即隐性感染(亚临床感染)。     

小鼠鼻气道阻力检测方法的建立

目的建立简单易行的小鼠鼻气道阻力检测方法及鼻灌洗方法。方法雌性小鼠16只,随机分为两组,每组8只,变应性鼻炎组（AR组）采用卵清蛋白＋氢氧化铝生理盐水混悬液腹腔注射致敏及滴鼻激发致敏,建立变应性鼻炎小鼠模型,对照组用生理盐水腹腔注射致敏和滴鼻激发。致敏后小鼠麻醉分离气管,近喉部切开气管插入18 G插管至鼻咽部,插管连接＂T＂形管,再分别连接实验精密注射泵和压力传感器,应用灌注/吸引模型模拟呼气/吸气,检测鼻气道的压力,计算鼻阻力。对小鼠鼻腔进行组胺雾化激发,检测鼻气道反应性。采用一侧鼻腔插管灌洗,对侧鼻腔收集灌洗液的方法,对鼻腔灌洗液进行中性粒细胞、嗜酸性粒细胞计数;处死小鼠后取鼻黏膜组织进行病理组织学观察。结果对照组小鼠的基础鼻气道阻力、反应性及鼻激发后的鼻气道阻力、反应性均低于AR组（P均〈0.01）;AR组鼻腔灌洗液中性粒细胞、嗜酸性粒细胞、细胞总数均显著高于对照组（P均〈0.01）;AR组的鼻黏膜有明显的嗜酸性粒细胞浸润,血管轻度充血水肿。对照组小鼠鼻黏膜无明显异常。结论成功建立了简单易行的小鼠鼻气道阻力检测方法及鼻灌洗方法。     

喷昔洛韦体内抗单纯疱疹病毒的作用

目的 :研究喷昔洛韦 (PCV)对单纯疱疹病毒 (HSV)感染小鼠的疗效。方法 :每只分别用病毒滴度为 10 0LD50 ·(0 .1ml) - 1 HSV 1、HSV 2腹腔注射 0 .0 3ml,2 4h后皮下注射给药 ,PCV剂量分别为 2 0 ,5 0 ,10 0 ,2 5 0mg·kg- 1 ·d- 1 ,疗程 5d。观察动物的反应 ,出现死亡时间和死亡数 ,所有动物用药后观察 15d。同时收集各组动物治疗后的标本 ,进行病毒分离 ,按常规测定病毒滴度。另设病毒对照组、0 .9%NaCI对照组、阳性药阿昔洛韦对照组 (ACV)。结果 :PCV对HSV感染鼠有保护作用 ,5 0mg·kg- 1 ·d- 1 时 ,HSV 1和HSV 2感染鼠的存活率分别为 6 0 %和 75 % ,平均存活时间 (d)分别为 11.3± 3.6和 12 .9± 2 .8,随着药物剂量增加 ,疗效增强 ,该药还能降低组织内病毒滴度 ,阻止体内病毒复制 ,抗HSV作用类似于相同剂量的ACV。结论 :PCV有抗HSV作用。     

Terminal dissemination of rabies virus in selected rat tissues.

Fifty rats were divided into 3 groups and challenged via the foot pad route with a fixed and 2 different strains of street rabies virus in order to study the dissemination of the virus and the affinity for certain tissues of the rat.The incubation period for rats inoculated with fixed rabies is shorter than with street virus, being 5 to 7 days compared with 10 to 12 days.Rats inoculated with the fixed strain were less aggressive and irritable than rats inoculated with street virus.The fixed strain demonstrated a greater affinity for the tissues studied as compared to the street strains of virus.Both the fixed and street strains revealed a low affinity for the parotid gland since no virus could be demonstrated in 14 of 20 in the fixed virus group and 27 of 30 in the street virus group.     

Laboratory studies on the transmission of western equine encephalitis virus by Saskatchewan mosquitoes. I. Culex tarsalis

A Saskatchewan strain of the mosquito Culex tarsalis, transmitted a local strain of western equine encephalitis virus from chick to chick, between four and 44 days after an infective blood meal. At incubation temperatures of 69 and 75 degrees F, 120 transmissions occurred out of a possible 141, and all but seven of these were by single infected mosquitoes. At 75 degrees F virus titers in individual mosquitoes were more uniform and transmission was more efficient, than at 69 degrees F, although infection rates were similar at both temperatures. The minimum concentration of virus required to infect 50% of C.tarsalis was 10(2.5) intracerebral three-week old mouse LD(50) per 0.03 ml of donor blood. These findings provide direct evidence that C. tarsalis of Saskatchewan is a highly efficient vector of western equine encephalitis virus.     

不同品系小鼠感染甲型流感病毒肺小血管内血栓形成

目的本实验旨在观察不同品系小鼠感染甲型流感病毒后肺组织内血栓形成的情况。方法使用H1N1病毒A/California/7/2009(CA7)株和H3N2病毒A/Brisbane/10/07株,对BALB/C小鼠、Scid小鼠、NOD/LTJ小鼠、BALB/C-nu小鼠、NOD-Scid小鼠和icosl-KO小鼠经乙醚麻醉后进行滴鼻攻毒。检测小鼠感染后肺组织病毒拷贝数并观察肺组织病理学改变。结果 H1N1和H3N2滴鼻攻毒的各组小鼠均染毒,病理表现为程度略有差异的间质性肺炎。13只H1N1病毒感染小鼠和6只H3N2感染小鼠在肺组织中观察到多个小血管内有血栓形成,血栓成分主要为纤维素和血小板。结论各品系小鼠感染H1N1和H3N2流感病毒后均可能出现肺组织内血栓形成。     

甲型H1N1流感病毒感染不同免疫缺陷小鼠的比较

目的宿主免疫系统的功能状态在病毒的感染中起着至关重要的作用,本实验观察了不同免疫缺陷小鼠感染甲型H1N1流感病毒的差异。方法使用六个品系的近交系小鼠,经乙醚麻醉后进行滴鼻攻毒,分析其在病毒感染后存活率、体重变化和肺组织病理改变的异同。结果感染H1N1病毒的6种小鼠在观察的14 d内,野生型的C57BL/6小鼠感染开始体重缓慢下降,感染后期有所回升,有半数存活;BALB/c小鼠和四种免疫缺陷品系小鼠感染病毒后体重随病情发展快速下降,死亡率均为100%。野生型C57BL/6小鼠感染初期为较弥漫的间质性肺炎,后期病变逐渐局限;BALB/c小鼠和四种免疫缺陷品系小鼠感染病毒后出现弥漫的中重度间质性肺炎,细支气管上皮有变性坏死,但炎症细胞明显少于C57BL/6小鼠。结论在甲型H1N1流感病毒的初次感染中固有免疫和特异性免疫分别在感染的初期和后期起主要作用,宿主免疫系统的功能状态影响着甲型H1N1病毒感染和预后。     

安徽省阜阳市狂犬病街毒株G基因序列分析

目的了解安徽省阜阳市流行的狂犬病毒株与人用、兽用狂犬病疫苗株在G基因核苷酸和氨基酸水平的差异,为有效控制我国狂犬病疫情提供初步科学依据。方法在安徽省阜阳市收集犬脑组织162份,用酶联免疫吸附试验（ELISA）和小鼠颅内接种试验（MIT）检测样品带毒情况,对阳性样品用RT—PCR扩增G基因并测序,以TOPALi和DNAStar软件对G基因序列进行分析。结果从安徽省阜阳市162份犬脑样品中检出阳性样品15份;这15株病毒与我国现在使用的各种疫苗株在G基因的核苷酸和氨基酸水平上均存在不同程度的变异,与我国人用疫苗株CTN同源性较高。结论15株狂犬病毒为基因Ⅰ型狂犬病毒,在核苷酸或氨基酸水平上,与疫苗株CTN之闻的同源性要高于与其它疫苗株之间的同源性。     

MSI和HPV16在鼻腔鼻窦肿瘤组织中的作用

目的初步探讨微卫星不稳定性(MSI)和16型人乳头状瘤病毒(HPV16)在鼻腔鼻窦肿瘤发生过程中的作用。方法应用聚合酶链式反应、变性聚丙烯酰胺凝胶电泳、银染等方法,检测57例鼻窦鼻腔肿瘤组织MSI的情况,采用免疫组织化学对HPV16进行检测。结果HPV16抗原表达的阳性细胞在凹空细胞处分布较多,恶性肿瘤组HPV16感染率(28.0%)低于IP组(58.3%)和IP伴鳞状上皮不典型增生(62.5%)组(P<0.05);IP伴鳞状上皮不典型增生组HPV16感染率与IP组比较无显著差异(P>0.05);空白对照组无阳性表达。恶性肿瘤组MSI的阳性表现率(36.0%)高于内翻性乳头状瘤组(0),差异有显著性(P<0.01);IP伴鳞状上皮不典型增生组MSI的阳性表现率(12.5%)低于恶性肿瘤组(36.0%),差异有显著性(P<0.05);IP伴鳞状上皮不典型增生组MSI的阳性表现率高于内翻性乳头状瘤组(P<0.05)。结论HPV16可能是鼻窦鼻腔肿瘤的重要致病因素,MSI亦可能与鼻窦鼻腔恶性肿瘤的发生密切相关,但HPV16可能不是通过MSI诱发鼻窦鼻腔恶性肿瘤的发生。