SYBR Green Ⅰ实时荧光定量RT-PCR测定猴免疫缺陷病毒RNA拷贝数方法的建立

.学者论坛·在动物实验中解决临床难题···························，·······································································……顾玉东(3)基因〕:程     

The comparative evaluation of cellular localization of viral antigens with microscopic changes in the ileum of cattle infected with bovine viral diarrhea

The correlation between microscopic changes with cellular localization of viral antigens was studied in the ileum of 16 cases infected with bovine viral diarrhea virus (BVDV). Microscopic lesions in the ileum included multifocal erosive and ulcerative ileitis, severe congestion and hemorrhage, crypt dilation and mucus engorgement, epithelial debris and leukocytes, lymphoid depletion of Peyer’s patches, herniation of mucosal epithelium into depleted Peyer’s patches, and fibrinoid vasculitis of submucosal vessels. BVDV antigen was detected by immunohistochemistry in macrophages, dendritic cells, smooth muscle cells, endothelial cells, epithelial cells of crypts, and mucosal epithelium, together with other mononuclear cells including lymphocytes, plasma cells, fibroblasts, and intramural ganglial cells. No consistent correlation between the presence of BVDV antigen and vascular lesions in the ileum was identified. The intensity and distribution of the immunoperoxidase stain in the ileum was graded as highly positive (18.7%), moderately positive (56.3%), and mildly positive (25%). In conclusion, the pattern and density of distribution and localization of BVDV antigen in the ileum was not consistently correlated with the severity of microscopic lesions.     

载氧血红蛋白纯化工艺研究

目的改善心脑血管缺氧载氧药物是一种创新药物。由于它半径比红细胞小400~1 000倍,易于通过毛细血管,给缺血组织及时供氧,迅速缓解或纠正缺氧状态,达到治疗抢救目的。血红蛋白的纯化工艺是载氧药物研制的重要工艺步骤。方法本研究建立了一套通过热敏法分离纯化人脐带血血红蛋白的工艺以及较为完善的纯化血红蛋白质量检测指标。结果与现有的纯化方式相比,热敏法操作简便,仪器设备造价低廉,纯化与病毒灭活同时进行,得到的纯化产品损失少,纯度高,各项理化指标达到国际水平。结论本工艺适用于规模制备纯化血红蛋白,为进一步研制治疗心脑血管缺氧载氧药物创造了有利条件。     

病毒感染与小儿短暂性滑膜炎关系的研究

探讨小儿短暂性滑膜炎与病毒感染 ,特别是柯萨奇B组病毒 (CVB)感染的关系。方法 :采用酶联免疫吸附试验 (ELISA)对 16 2例小儿髋关节短暂性滑膜炎患儿静脉血和其中的 2 1例关节腔液进行病毒IgM抗体检测 ,柯萨奇病毒B组 (CVB)阳性者进行血清学分型及病毒分离。结果 :在 16 2例静脉血样本中病毒IgM阳性 6 2例 ,占总检数的 38.2 7% ,CVB -IgM2 6例占 16 .0 5 % ,AdV -IgM 19例占 11.73% ,CMV -IgM 10例占 6 .17% ,RSV -IgM 4例占 2 .4 7% ;在关节腔液中检出 9例CVB阳性 ,血清学分型 8例为CVB3,1例为CVB4 ,并分离出 1例CVB3。结论 :CVB和腺病毒 (AdV)是小儿髋关节短暂性滑膜炎感染的主要病毒 ,而且以CVB3为主。     

病毒性心肌炎心肌肌钙蛋白Ⅰ检测的临床意义

目的:对病毒性心肌炎患儿血中肌钙蛋白I(cTn I)含量进行检测,探讨与临床的关系。方法:采用放射免疫法对24例病毒性心肌炎患儿血中cTn I进行定量检测,同时检测血清心肌酶谱的改变情况,并就两种指标进行相关性探讨。结果:病毒性心肌炎患儿血中cTn I含量明显增高(4.5±2.2 )vs(2.3±0.3)ng/ml,P<0.01),并与CKMB,LDH_1的含量呈明显的正相关。结论:cTnl I是诊断病毒性心肌炎的敏感指标,可以反映出心肌受损的程度。     

乙型肝炎甲胎蛋白检测与重型肝炎预后指数评估的意义

给慢性活动肝炎患者输注人胎肝细胞悬液引起血清甲胎蛋白增高反应的初浓度为２０２±３８．３μｇ／Ｌ，其半衰期为１．７天；分布池为６．９一１０．２升。重型肝炎生存组血清甲胎蛋白显著高于死亡组，生存组甲胎蛋白与血清总胆红素呈非常显著相关，而死亡组两者无相关性。重型肝炎患者输注胎肝悬液后其血清甲胎蛋白进一步升高者，预后较好。取重型肝炎预后指数０．１为重型肝炎生死的截断点，判断预后的灵敏度、特异度、准确度和阳性预告值分别为７９．５％、８９．６％、８４．８％和８７．５％。重型肝炎预后指数的计算可作为判断预后与新疗法疗效的指标。     

Human spumaretrovirus antibody reactivity in multiple sclerosis

The role of human spumaretrovirus (HSRV) infections in the pathogenesis of multiple sclerosis (MS) was investigated with recombinant HSRV env-specific enzyme-linked immunosorbent assay. The presence of HSRV antibodies was determined in pairs of serum and cerebrospinal fluid (CSF) samples from 60 MS patients. In 7 of these patients serial serum and CSF samples were obtained in relation to the clinical activity of the disease during a period of 2 years. No increased antibody reactivity was demonstrable in the MS population compared with 14 aseptic meningitis patients, 50 blood donors and 16 healthy controls. Slightly elevated levels of antibodies were demonstrable in serum and/or CSF in 4 MS patients but also in 1 patient with aseptic meningitis, 1 blood donor and 1 child. No marked serum or CSF HSRV antibody fluctuation was observed in the MS patients followed longitudinally. Thus, this study does not support the involvement of HSRV in the pathogenesis of MS.     

病毒性心肌炎心肌线粒体结构功能变化及大剂量维生素C干预作用

目的：探讨病毒性心肌炎（ＶＭＣ）小鼠心肌线粒体结构和功能，并用大剂量维生素Ｃ进行干预。方法：雄性Ｂａｌｂ／ｃ小鼠随机分为柯萨奇Ｂ３病毒（ＣＶＢ３）感染组（ＩＧ）、ＣＶＢ３感染加大剂量维生素Ｃ治疗组（ＩＶＧ）和对照组（ＣＧ）。采用电镜和形态计量学方法观察心肌线粒体形态、数量和膜磷脂定位，用酶细胞化学法分析心肌线粒体细胞色素氧化酶（ＣＣＯ）和琥珀酸脱氢酶（ＳＤＨ）活性。结果：ＩＧ小鼠心肌线粒体大量破坏，ＣＣＯ和ＳＤＨ活性降低，膜磷脂严重缺失、定位改变（Ｐ　＜０．０５～　０．０１）。不同时相点ＩＶＧ上述各项指标均较ＩＧ显著改善（Ｐ　＜０．０５～０．０１）。结论：ＶＭＣ心肌线粒体结构严重破坏、功能明显下降，大剂量维生素Ｃ能有效保护心肌线粒体结构和功能。     

A macrophage inflammatory protein homolog encoded by guinea pig cytomegalovirus signals via CC chemokine receptor 1

Cytomegaloviruses encode homologs of cellular immune effector proteins, including chemokines (CKs) and CK receptor-like G protein-coupled receptors (GPCRs). Sequence of the guinea pig cytomegalovirus (GPCMV) genome identified an open reading frame (ORF) which predicted a 101 amino acid (aa) protein with homology to the macrophage inflammatory protein (MIP) subfamily of CC (β) CKs, designated GPCMV-MIP. To assess functionality of this CK, recombinant GPCMV-MIP was expressed in HEK293 cells and assayed for its ability to bind to and functionally interact with a variety of GPCRs. Specific signaling was observed with the hCCR1 receptor, which could be blocked with hMIP −1α in competition experiments. Migration assays revealed that GPCMV-MIP was able to induce chemotaxis in hCCR1-L1.2 cells. Antisera raised against a GST-MIP fusion protein immunoprecipitated species of ∼12 and 10 kDa from GPCMV-inoculated tissue culture lysates, and convalescent antiserum from GPCMV-infected animals was immunoreactive with GST-MIP by ELISA assay. These results represent the first substantive in vitro characterization of a functional CC CK encoded by a cytomegalovirus.