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针对当前很多医院处于高负荷运转状态,内涵质量建设存在不少薄弱环节的实际,创新提出以“三个核心”为突破口,聚焦质量建设基础,狠抓“核心制度”落实,紧扣质量建设主旨,强化“核心指标”监管,扭住质量建设关键,深挖“核心员工”潜能,不断提升内涵质量建设水平,夯实可持续发展根基,为推进医院质量建设开拓了新的思路,为医院可持续发展奠定了基础。  相似文献   

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杜勇  周建军 《病毒学报》1998,14(4):307-314
设计了利用大扬杆菌鞭毛蛋白递呈的随机十二肽库研究HCV核心蛋白B细胞抗原位的实验程序:1利用大肠杆有达质粒pQE-30有达并纯化HCV核心蛋白P19;2利用P19蛋白亲和层析纯化HCV感染者血清的抗HCV核心蛋白多克隆抗体;  相似文献   

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大量的跨膜受体能靶向并定位于细胞核内, 但细胞表面的受体是如何转运到细胞核内尚不清楚. 报道了磷酸化的TrkA在人脑胶质瘤细胞株U251中的定位. 利用免疫细胞化学和免疫荧光技术, 发现磷酸化的TrkA主要定位于一系列的运输泡和细胞核内, 这些膜泡包括: 细胞膜侧的环状泡、核周的大核心泡和小核心泡; 同时还观察到大核心泡出芽成小核心泡, 以及小核心泡与核膜相互作用. 基于上述结果认为这些膜泡可能与跨膜受体TrkA转位到核的通路相关.  相似文献   

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HCV核心区与HBV核心区融合基因的DNA免疫   总被引:6,自引:0,他引:6  
构建了在CMV启动子控制下在真核系统表达丙肝核心蛋白基因全长(HCc1 91 )及其氨端片段 (HCc6 9及HCc40 )的表达克隆 ,以及这 3种不同长度的HCc分别与乙肝核心区基因 (HBc1 44 )在羧端融合的表达克隆 .在COS细胞中实现了暂时表达 .ELISA和Westernblot分析表明 ,表达产物具HCc抗原性或同时具有HBc的抗原性 .非融合和融合基因的DNA直接免疫小鼠 ,能有效地产生抗HCc抗体或同时产生抗HBc抗体 .融合形式要比非融合形式产生抗HCc抗体的持续时间长 .结果表明不同长度的HCc的融合并不影响HBc免疫原性的表现 ,却有利于HCc免疫原性的表现 .  相似文献   

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为提高种质资源在育种中的利用效率,建立了我国绿豆(Vigna radiata)应用型核心样本。该样本既包括了资源库中具特异性状的种质和曾经在生产上大面积种植的品种,也包括了在育种中使用频繁的亲本及苗头品系等。农艺性状变异分析表明,该核心样本具有丰富的表型变异,是绿豆种质资源的代表性样本。聚类分析可将核心样本分为4大类,但类别内种质与其地理来源相关不明显。不同来源表型数据的分析发现,不同性状间的一致性存在差异。但产量相关性状的表现均与当前育种目标相接近,说明该核心样本具有较高的实用性。  相似文献   

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肠杆菌基因间重复共有序列及ERIC-PCR   总被引:6,自引:0,他引:6  
ERIC序列是近年来发现的存在于原核生物基因组中一类短的重复序列。该序列在染色体上的分布和拷贝数具种间特异性,根据序列中心高度保守的44bp的ERIC核心序列设计反向引物,可扩增出反映细菌基因组结构特征的谱带。由于ERIC-PCR快速简便,图谱重复性好,并可作为分子标记用于细菌的分类鉴定,所以,该方法已广泛应用到了科研和生产实践中;  相似文献   

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目的探讨不同种雏鸭建立鸭乙肝病毒感染模型的影响因素,观察应用该模型抗病毒的效果。方法采集鸭血清,应用PCR方法定性检测鸭血清中病毒DNA;定量PCR方法检测鸭血清中病毒DNA载量变化;用抗病毒药物处理,观察其在鸭DHBV感染模型中的抗病毒效果。结果不同种鸭DHBV自然感染率不同,樱桃谷鸭为8.75%,湖北麻鸭两个批次分别为17.80%和10.68%;静脉注射和腹腔注射两途径均能致雏鸭感染DHBV,静脉注射感染率80%,腹腔注射感染率65%;鸭感染DHBV后,体内病毒载量维持在106~108copies/mL,可持续20 d以上;抗病毒药物处理后,在不同DHBV模型中其抗病毒效果变化趋势一致。结论鸭的种类和人工感染途径可影响DHBV感染率;雏鸭感染DHBV后其体内有持续性的病毒血症;DHBV感染模型是药物抗病毒研究较好模型。  相似文献   

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鸭乙型肝炎病毒前S抗原决定簇的研究   总被引:1,自引:0,他引:1  
闻玉梅  刘寅曾 《病毒学报》1990,6(2):145-150
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T Saito  K Tachibana  K Mogi  H Mizuo  Y Ito  M Imai 《Uirusu》1989,39(1):55-60
Mice were immunized against duck hepatitis B virus core (DHBc) particles isolated from the liver of asymptomatic carrier ducks of duck hepatitis B virus (DHBV) by ultracentrifugation. Their spleen cells were fused with mouse myeloma (NS-1) cells, and 12 clones of hybridoma cells secreting antibodies against DHBc (anti-DHBc) were isolated. According to the reactivity to core particles and core peptide obtained from DHBc particles treated with SDS-2ME, the 12 antibodies were classified into two groups. Two monoclonal antibodies reacted against both core particles and core peptide (B-type), the other ten monoclonal antibodies reacted against core particles but did not react against core peptide obtained from DHBc particles treated with SDS-2 ME. (A-type). Solid phase enzyme immuno assay (EIA) using these two types of antibodies could detect core antigenisity not only in the liver homogenate but also in the DHBV infected serum. Sucrose gradient analysis and gel filtration analysis revealed this DHBc antigenisity in the serum is not carried by core particles but carried by core peptide, equivalent to HBe antigen in the serum of Hepatitis B virus (HBV) carrier. This EIA may provide sensitive test monitoring both serum DHBe antigen levels and DHBc antigen levels in the liver during DHBV infection.  相似文献   

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The induction of humoral response in ducks by DNA-based immunization against duck hepatitis B virus (DHBV) core protein (DHBc) was investigated. In addition, the amino acid specificity of the induced response was compared by using peptide scanning to that elicited either by protein immunization or during chronic DHBV infection. Immunization of ducks with a plasmid expressing DHBc protein led to the induction of a long-lasting antibody response able to specifically recognize viral protein in chronically infected duck livers. Peptide scanning analysis of anti-DHBc response induced during chronic DHBV infection allowed us to identify six major antigenic regions (AR1 to AR6). The reactivity spectrum of duck sera elicited by protein immunization appeared narrower and was restricted to only four of these antigenic regions in spite of higher anti-DHBc antibody titers. Interestingly, anti-DHBc antibodies induced by DNA-based immunization recognized five of six antigenic regions, and the epitope pattern was broader and more closely related to that observed in chronic viral infections. To gain more insight into the location of antigenic regions, we built a three-dimensional (3-D) model of DHBc protein based on human and duck core sequence alignment data and the HBc 3-D crystal structure. The results suggest that two identified antigenic regions (AR2, amino acids [aa] (64)T-P(84), and AR5, aa (183)A-R(210)) are located at positions on the protein surface equivalent to those of the two HBc major epitopes. Moreover, we identified another antigenic region (AR3, aa (99)I-I(112)) that was recognized by all sera from chronically infected, DNA- or protein-immunized ducks within the large 45-aa insertion in DHBc protein, suggesting that this region, which lacks HBc, is externally exposed.  相似文献   

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Antisense therapy of hepatitis B virus infection   总被引:2,自引:0,他引:2  
Chronic infection with the hepatitis B virus (HBV) is a major health problem worldwide. The only established therapy is interferon-a with an efficacy of only 30–40% in highly selected patients. The discovery of animal viruses closely related to the HBV has contributed to active research on antiviral therapy of chronic hepatitis B. The animal model tested and described in this article are Peking ducks infected with the duck hepatitis B virus (DHBV). Molecular therapeutic strategies aimed at blocking gene expression include antisense DNA. An antisense oligodeoxynucleotide directed against the 5′-region of the preS gene of DHBV inhibited viral replication and gene expression in vitro in primary duck hepatocytes and in vivo in Peking ducks. These results demonstrate the potential clinical use of antisense DNA as antiviral therapeutics.  相似文献   

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鸭乙型肝炎病毒实验感染后在外周血和肝脏中的动态   总被引:3,自引:0,他引:3  
鸭乙型肝炎病毒(DHBV)与人乙型肝炎病毒(HBV)同属嗜肝病毒,两者的病毒大分子结构和复制过程有很多相似之处,了解DHBV实验感染规律和病毒在血液和肝脏内的动态,有助于研究病毒复制特点和抗肝炎药物的效果。 本文用DHBV-DNA杂交阳性和DHBV-DNA多聚酶阳性的上海鸭血清,静脉注射1~3  相似文献   

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选取3份DHBV阳性广东麻鸭血清提取DHBVDNA,设计一对扩增DHBV全基因序列引物Q1和Q2,扩增并克隆DHBV全基因序列,测序并运用相关计算机软件及方法分析获得的全基因序列。结果表明,广东麻鸭DH-BV全长为3027bp。提交GenBank后获得的收录号分别为:AY433937、AY521226、AY521227(来自1号血清);AY392760、AY536371(分别来自2、3号血清)。AY521227的PreS/SORF出现了单碱基突变,在PreC/CORF之前发现一个新的ORF,暂命名为HORF,HORF也同时存在于GenBank中储存的另外8个DHBV全基因序列中。系统发育分析表明,广东麻鸭DHBV在分化程度上是目前储存于GenBank中的DHBV全序列中最高的。成功克隆了广东麻鸭DHBV全基因序列,序列分析为进一步研究广东麻鸭DHBV提供了有益的信息。  相似文献   

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为建立鸭乙型肝炎病毒LJ-76的转染细胞系,将LJ-76病毒DNA插入到pUC19的EcoRⅠ位点上,分离得到含有双拷贝LJ-76DNA的重组质粒.通过磷酸钙沉淀方法,将经CsCl等密度离心纯化的LJ-76DNA双体导入到人肝癌细胞BEL7402中.收集转染细胞的培养液进行蔗糖密度梯度离心,所得沉淀经检测发现含有LJ-76DNA并具有特异性DHBV内源性DNA多聚酶活性;对上述样品通过DotEIA检测DHBV核心抗原及表面抗原结果为阳性.Southernblot分析表明转染细胞内存在病毒DNA复制中间体cccDNA、ssDNA和rcDNA,而cccDNA被认为是复制活动较为活跃的标志.电镜观察转染细胞的上清发现有病毒颗粒的存在.  相似文献   

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用鸭乙型肝炎病毒(DHBV)阳性的安徽庐江鸭血清感染DHBV阴性的北京雏鸭,扩增病毒,将提取的DHBV-DNA插入pUC18质粒,转化E.coli JM 105。酶切重组质粒及South-ern转膜杂交结果证实,质粒pLJ76的插入片段为DHBV全基因组。用EcoR Ⅰ等11种限制性内切酶对pLJ76进行酶谱分析,并与美国,西德的已知DHBV基因组比较。定向克隆该株病毒不同基因编码区片段,构建正负单链探针,将斑点杂交和单链电泳检出的M13阳性重组子与已知序列的DHBV基因组作比较,提示获得了该株病毒基因组的S、Pre-S、P和X/C等蛋白编码区的正、负单链克隆株。  相似文献   

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