肾综合征出血热病毒结构蛋白致病作用的研究

为阐明肾综合征出血热（ＨＦＲＳ）发病机制，用免疫组化法检查了３０例患者ＨＦＲＳ病毒的膜蛋白（ＭＰ）、核蛋白（ＮＰ）在外周血单个核细胞（ＰＢＭＣ）中的表达情况，同时用放免等技术检测了血浆内皮素（ＥＴ）、Ｐ物质（ＳＰ）和肾功能相关指标。发现ＨＦＲＳ患者从４～５病日起至１３病日，ＰＢＭＣ中均有ＭＰ和ＮＰ表达，但随着病情的好转，其表达强度逐渐减弱。经染色鉴定ＭＰ与ＮＰ阳性细胞主要是单核细胞。入院时ＰＢＭＣ中ＭＰ和ＮＰ表达强度并不相同，ＭＰ的表达强度与血浆ＥＴ／ＳＰ比值、病情及肾脏损伤轻重呈平行关系。提示ＨＦＲＳ病毒能侵犯ＰＢＭＣ并在其中复制，表达病毒结构蛋白，而ＭＰ与ＨＦＲＳ病毒的致病作用相关；病程中血浆ＥＴ／ＳＰ比值升高是使肾损加重的重要因素。     

恶性疟原虫裂殖子表面蛋白1-17区基因的原核表达、纯化及表达产物鉴定

目的　为进一步研究和应用恶性疟原虫裂殖子表面蛋白１ 的１７区基因（ＭＳＰ１－ １７）,本文原核表达了ＦＵＰ株ＭＳＰ１ 的１７ 区基因,并纯化及鉴定了表达蛋白。方法　将ＭＳＰ１－ １７ 基因片段克隆入原核表达载体ｐＧＥＸ－ ４Ｔ－ １,形成ｐＧＥＸ－ ４Ｔ－ １／ＭＳＰ１－ １７。ＩＰＴＧ诱导ｐＧＥＸ－ ４Ｔ－ １／ＭＳＰ１－ １７ 转化的ＢＬ２１菌,纯化并用ＭＳＰ１－ １７ 特异性单克隆抗体鉴定表达产物。结果　成功地构建了ｐＧＥＸ－ ４Ｔ－ １／ＭＳＰ１－ １７,转化菌经诱导表达出与ＧＳＴ融合的蛋白（约４０ＫＤ）,纯化后纯度达７２％ ,ＭＳＰ１－ １７ 特异性单克隆抗体可识别表达蛋白。结论　成功表达并纯化了ＭＳＰ１－ １７ 蛋白,为深入研究和应用ＭＳＰ１－ １７ 打下基础     

流行性出血热并发神经系统损害机理的临床研究

对４６例流行性出血热（ＥＨＦ）伴神经系统（ＮＳ）损害者进行了临床和实验室研究，结果表明，ＥＨＦ伴ＮＳ损害的病因以ＥＨＦ脑炎最多。根据１例患者的脑脊液（ＣＳＦ）中分离到ＥＨＦ病毒（ＥＨＦＶ），９３．１％患者ＣＳＦ中抗－ＥＨＦＶ－ＩｇＭ阳性，而无ＮＳ症状的对照组全部阴性等结果，认为ＥＨＦＶ的直接致病作用为ＥＨＦ脑炎的主要病因，提示早期抗病病毒治疗可能预防ＥＨＦ脑炎。低蛋白血症是脑水肿、脑疝的原因，及时     

病毒胸苷激酶基因治疗人胃癌的实验研究

为观察单纯疱疹病毒脱氧胸苷激酶（ＨＳＶ－ＴＫ）／丙氧鸟苷（ＧＣＶ）基因治疗系统对胃癌细胞的杀伤作用，将ＨＳＶ－ＴＫｃＤＮＡ定向克隆入逆转录病毒载体ｐＤＯＲ－ｎｅｏ中，用ＬＩＰＯＦＥＣＴＡＭＩＮＥ法将该ＨＳＶ－ＴＫ基因转染胃癌细胞系ＳＧＣ－７９０１，测定阳性转染细胞（ＳＧＣ－７９０１／ＴＫ）在体内外对ＧＣＶ的敏感性。结果示ＧＣＶ在体外对ＳＧＣ－７９０１／ＴＫ细胞有明显的杀伤作用，其作用呈现剂量和时间依赖性特点，且同时表现出对周围亲代ＳＧＣ－７９０１细胞的杀伤效应（旁观者效应）。体内实验表明ＧＣＶ可抑制ＨＳＶ－ＴＫ阳性胃癌细胞的生长，使已形成的肿瘤逐渐消失。提示逆转录病毒载体介导的ＨＳＶ－ＴＫ基因转移胃癌细胞，配合以ＧＣＶ治疗，是胃癌基因治疗的又一途径。     

抗结核药物联合使用对耐多药结核分枝杆菌抗菌效力的研究

目的 研究常用抗结核药物与其它抗结核药物联合使用对多耐药结核菌（ＭＤＲ－ＭＴＢ）的抗菌效力。方法 按照部分耐药浓度（ＦＩＣ）方法测定（ＭＤＲ－ＭＴＢ）单独用药及联合用药时标准结核分枝杆菌的最低抑菌浓度（ＭＩＣ）,临床分离株的最低抑菌浓度（ＭＩＣｓ）,按照ＦＩＣ公式,计算其是否有协同作用。结果 ＩＮＨ、ＲＦＰ、ＥＭＢ、ＣＡＭ和ＩＮＨ、ＲＦＰ、ＰＺＡ、ＯＦＬＸ组合对１３株临床分离的ＭＤＲ－ＭＴＢ的ＦＩ     

恶性疟原虫裂殖子表面蛋白1—17区基因的原核表达,纯化及表？…

为进一步研究和应用恶性疟原虫裂殖子表面蛋白１的１７区基因,本文原核表达了ＦＵＰ株ＭＳＰ１的１７区基因,并纯化及鉴定了表达蛋白。方法将ＭＳＰ１－１７基因片段克隆入原核表达载体ｐＧＥＸ－４Ｔ－１,形成ｐＧＥＸ－４Ｔ－１／ＭＳＰ１－１７,ＩＰＴＧ诱导ｐＧＥＸ－４５－１／ＭＳＰ１－１转化的ＢＬ２１菌,纯化并用ＭＳＰ１－１７特异性生有达产物。结果成功地构建了ｐＧＥＸ－４Ｔ－１／ＭＳＰ１－１７,转化菌经诱导表     

弓形虫病诊断方法的对比研究

应用５种不同方法测定了６例弓形虫病人不同病期的血清标本。用ＩＨＡＴ和ＩＦＡＴ测定弓形虫总免疫球蛋白，ＥＨＡＴ在发病后第３周的２份血清标本中有１份已呈阳性，１４－１７周达高峰。滴度升高非常明显；ＩＦＡＴ第３周的标本都呈阳性；１７－２４周达高峰。用ＩｇＧ－ＥＬＩＳＡ和ＩｇＭ－ＥＬＩＳＡ，ＩｇＭ－ＩＦＡＴ分别测定抗弓形虫特异性抗体ＩｇＧ和ＩｇＭ＜ＩｇＧ－ＥＬＩＳ从第６周呈阳性，随病情延长，滴度明显升高，     

9株艾滋病病毒的分离及生物学特性的研究

目的 从艾滋病病毒（ＨＩＶ）感染者血液中分离ＨＩＶ,进行ＨＩＶ分离株的生物学特征研究。方法 采集１０份福建艾滋病病毒感染者肝素抗凝血,分离外周血单核细胞（ＰＢＭＣ）,与健康人ＰＢＭＣ共培养进行ＨＩＶ－１的分离,使用含神经氨酸酶（ＮＡ）的Ｔ细胞培养液提高病毒分离率,通过检测Ｐ２４抗原、间接免疫荧光试验（ＩＦＡ）及电镜观察等确定病原分离结果。用Ｈ９及ＭＴ４细胞对分离的病毒进行细胞嗜性的研究。结果 从１     

抗肾综合征出血热病毒独特型人源性单克隆抗体的制备

分离肾综合征出血热（ＨＦＲＳ）恢复期患者外周静脉血淋巴细胞，经分离的Ｂ淋巴细胞ＥＢ病毒转化后分别与人─鼠种间骨髓瘤细胞系ＳＨＭ－Ｄ３３和小鼠骨髓瘤细胞系Ｘ６３－Ａｇ８．６５３融合，融合率分别为９６％和９０％。杂交瘤生长孔分泌抗ＨＦＲＳＶ抗体阳性率达３３％和２３％。而且均有一孔为抗ＨＦＲＳＶ独特型抗体阳性，经克隆化筛选各得到一株杂交瘤细胞系，命名为Ｃ８和Ｆ３。经鉴定两株杂交瘤细胞系分泌的人源性单克隆抗体均为ＩｇＭ型。Ｆ３抗体产量小于１ｍｇ／ｍｌ，Ｃ８抗体产量为３０～５０ｍｇ／ｍｌ，且已稳定传代两年以上。抗ＨＦＲＳＶ独特型人源性单克隆抗体的成功制备，为ＨＦＲＳＶ疫苗的研制开辟了一条新路。     

病毒性肝炎患者血清HGF和TGF—β1水平的临床意义

目的：探讨血清肝细胞生长因子（ｓＨＧＦ）和转化生长因子－β＿１（ＴＧＦ－β１）水平在病毒性肝炎患者中的临床意义。方法：选１３９例病毒性肝炎患者，以双抗体夹心ＥＬＩＳＡ检测ｓＨＧＦ，以改良ＭＶ１ＬＵ细胞生长抑制ＭＴＴ法检测ＴＧＦ－β＿１活性，同时检测其肝功能、肝纤维化、甲胎蛋白等指标。结果：各型病毒性肝炎患者ｓＨＧＦ水平及ＴＧＦ－β＿１活性均明显高于正常人水平（Ｐ值均＜０．０１）。ＴＧＦ－β１活性增高以肝硬化和重症肝炎最为显著（分别为９．４４±２．１７ｎｇ／ｍｌ和８．４２±２．５４ｎｇ／ｍｌ）。ｓＨＧＦ水平与ＴＧＦ－β＿１活性相关（Ｐ值＜０．０１）。结论：ｓＨＧＦ水平能反映患者肝细胞损伤及肝功能障碍程度，并与肝纤维化程度也可能有关。     

Crimean-congo hemorrhagic fever, Thogoto, dugbe, and Jos viruses isolated from ixodid ticks in Ethiopia.

From 1974 through 1976, U.S. Naval Medical Research Unit No. 5 isolated 25 strain of tick-borne virus in infant mice from 410 pools containing over 6,000 ticks, and one strain from a bird and one strain from a rodent collected in central and southern Ethiopia. Of these, 17 were identified as known viruses previously found in West Central and East Africa. There were 8 strains of Jos virus from Amblyomma ticks; 7 strains of Dugbe virus from a bird, a rodent and from ticks; 1 strains of Crimean-Congo hemorrhagic fever virus and 1 strain of Thogoto virus from ticks.     

首次从新疆的蜱和病牛血清分离到新环状病毒

1990年3～8月从新疆北部和西部地区采集全沟硬蜱(Ixodes persulcatus)、银盾革蜱(Dermacentor niveus)、森林革蜱(D.silvarum)、边缘革蜱(D.marginatus)、草原革蜱(D.nuttalli)、刻点血蜱(Haemaphysalis punctata)、残缘璃眼蜱(Hyalomma detritum)等共2132只,分离病毒17株;采集病牛血清3份,分离病毒1株。从两地区共分离病毒18株。经血清学鉴定,该类病毒与披膜病毒科(Togaviridae)、黄病毒科(Flavirdae)、布尼亚病毒科(Bunyavirdae)某些病毒的免疫腹水及免疫血清都不发生反应。与新环状病毒(BANNA)的免疫腹水、免疫血清及酶标单克隆抗体(McAb)发生反应,中和指数为4.22～5.03。电镜观察病毒为球形无囊膜颗粒,直径为63～69nm,核衣壳直径为49～54nm。超薄切片在感染细胞中见到大量颗粒状包涵体,磷钨酸负染可见到排列整齐的壳粒结构,显示了典型的环状病毒形态特征。新分离的病毒对酸、氯仿、胰蛋白酶及热敏感;抵抗乙醚和5-碘脱氧尿苷。该病毒可引起白纹伊蚊(Aedes albopictus)传代细胞 C_6l_(36)细胞收缩、融合、脱落及颗粒增多等病变;感染传代细胞Vero株及BHK-21株未见明显的细胞病变。感染2～4日龄乳小白鼠及2～4周龄小白鼠,均无明显发病症状。初步鉴定证实,新分离病毒属新环状病毒(BANNA)。     

Endemicity of spotted fever group rickettsiae in Connecticut

To compare rickettsial infectivity and seropositivity rates against spotted fever group (SFG) rickettsiae, ticks and wild mammals were collected from three areas where Rickettsia rickettsii was thought to be enzootic in Connecticut during 1978-1979, and from four additional sites (with no reported human cases) between 1976 and 1979. Of the 1,001 Dermacentor variabilis examined by the hemolymph test, 59 (5.9%) contained rickettsia-like organisms; direct immunofluorescence tests verified the presence of SFT rickettsiae in 14 specimens. Prevalence of rickettsiae-infected ticks at Newtown, an area where human cases of Rocky Mountain spotted fever probably originated, was 2.2%. Rates for six other areas ranged between 0 and 6.3%. Isolations included Rickettsia montana from four ticks collected at Branford and Woodbridge, and R. rickettsii (R-like strain) from the blood of an acutely ill person. Microagglutination (MA) tests indicated that 15 (14.9%) of 101 Peromyscus leucopus (white-footed mice) from Newtown had agglutinins in titers greater than or equal to 1:8 against R. rickettsii, whereas five of 92 white-footed mice (5.4%) from Brandord, West Hartford, Woodbridge, and Sharon were considered MA-positive. Indirect microimmunofluorescence tests of Procyon lotor (raccoon) sera revealed antibodies to R. rickettsii in 33 of 69 (47.8%) samples from Newtown and in two of 60 (3.3%) from Guilford. Additionally, 17 raccoons had sera specific to R. montana (n = 8) or to the 369-C rickettsia strain (n = 9). Since rickettsia-positive ticks are high-titered seropositive mammals occurred at widely separated sites in Connecticut there are probably several foci of SFG rickettsiae distributed throughout the D. variabilis range.     

West Nile and Usutu Virus Introduction via Migratory Birds: A Retrospective Analysis in Italy

The actual contribution of migratory birds in spreading West Nile (WNV) and Usutu virus (USUV) across Europe and from Africa to old countries is still controversial. In this study, we reported the results of molecular and serological surveys on migrating birds sampled during peaks of spring and autumn migration at 11 Italian sites located along important flyways, from 2012 to 2014. A total of 1335 specimens made of individual or pooled sera, and organs from 275 dead birds were tested for WNV and USUV RNA by real time PCR (RT-PCR). Furthermore, sera were tested by serum neutralization assay for detecting WNV and USUV neutralizing antibodies. Molecular tests detected WNV lineage 2 RNA in a pool made of three Song Thrush (Turdus philomelos) sera sampled in autumn, and lineage 1 in kidneys of six trans-Saharan birds sampled in spring. Neutralizing antibodies against WNV and USUV were found in 5.80% (n = 72; 17 bird species) and 0.32% (n = 4; 4 bird species) of the tested sera, respectively. Our results do not exclude the role of migratory birds as potential spreaders of WNV and USUV from Africa and Central Europe to Mediterranean areas and highlight the importance of a more extensive active surveillance of zoonotic viruses.     

云南首次分离到辛德毕斯(Sindbis)、巴泰(Batai)和Colti病毒

目的了解云南省虫媒病毒分布情况,为防治提供依据。方法在云南省思茅地区和西双版纳州采集蚊虫以及发热病人血清,液氮冻存。标本常规处理,接种C6/36细胞和乳鼠以分离病毒,并用血清学和分子生物学方法对分离到的病毒进行鉴定。同时采集发热病人和健康人血清,用ELISA和血凝抑制试验检测病毒抗体。结果从西双版纳发热病人血清中分离到1株辛德毕斯(Sindbis)病毒,从澜沧县菲律宾按蚊中分离到1株巴泰(Batai)病毒,从思茅市翠云区和澜沧县三带喙库蚊、中华按蚊、菲律宾按蚊、迷走按蚊等蚊虫中分离到10株Colti病毒。血清抗体检查,西双版纳发热病人血清中辛德毕斯病毒抗体阳性率为2.50%(3/120),巴泰病毒抗体阳性率为4.17%(5/120);思茅和西双版纳地区健康人血清中辛德毕斯病毒抗体阳性率为1.93%(11/571),其中澜沧、思茅、景洪、勐腊和勐海的阳性率依次为4.55%(6/132)、0.89%(1/112)、1.25%(2/160)、1.96%(2/102)和0.00%(0/65);西双版纳发热病人和脑炎病人血清中还检测出Colti病毒抗体。结论云南省分布有经蚊虫传播的辛德毕斯、巴泰和Colti病毒,当地人群中也存在该病毒自然感染。今后应加强这3种虫媒病毒病的调查研究和防治工作。     

陕西延安地区肾综合征出血热流行情况研究报告

目的 明确陕西延安地区为肾综合征出血热疫源地及病毒基因型别。方法 择有疫情报告地,进行人群疫情调查及健康人群特异性抗体检测;用夹夜法捕鼠,调查鼠种及鼠密度,采集鼠肺、血标本。人血、鼠血分别用IFA和ELISA法检测IgG抗体;鼠肺采用RT-PCR检测汉坦病毒(HV)RNA,测定PCR产物并与HV代表株76-118和Seoul序列进行比较。结果 对延安市防疫站2001年统计的7例HFRS患者进行流行病学调查,均属在当地感染;检测延安市洛川县交口河镇245例人血标本HV特异性IgG抗体,阳性50例,人群隐性感染率为20.41％。22份鼠肺及鼠血标本,IFA抗HV IgG阳性3例,ELISA阳性5例。从一只鼠血抗体阳性的鼠肺中检测到HV RNA,序列测定符合Seoul S片段序列(94％)。结论 延安市洛川县交口河镇为HFRS疫源地及新疫区,洛川交口河镇健康人群、褐家鼠、小家鼠中有HV感染,病毒型别为SEO型。     

The search for Ixodes dammini and Borrelia burgdorferi in Nova Scotia

Twenty-four Ixodes dammini ticks (23 adults and one nymph) have been recovered in Nova Scotia since 1984. There has not been a systematic search for larvae and none has been identified. The recovery of the nymph from a road-killed yellow throat bird, Geothypis trichas, in late May 1990 supports the contention that migrating birds are bringing deer ticks into the province every spring. In March and April 1991, four adult deer ticks were identified, suggesting that these ticks had overwintered. These deer tick specimens indicate that it is possible that I dammini is becoming established in Nova Scotia, if it is not already established. There has been no evidence for the existence of Borrelia burgdorferi in the province. The spirochete was not cultured from 650 Dermacentor variabilis ticks, nor were antibodies detected in a small sample of feral rodents using an indirect fluorescent antibody test. A survey of 137 dog sera samples, analyzed by enzyme-linked immunosorbent assay, also proved negative. There has been no confirmed indigenous case of Lyme disease in Nova Scotia to date.     

Interference between the viruses of tick-borne encephalitis complex in the Ixodes ricinus ticks.

The interference of two closely related viruses of the tick-borne encephalitis complex (TBE) in Ixodes ricinus ticks has been observed. 42.9 to 65.0% of dually infested ticks developed virus neutralization antibodies in mice without signs of illness, which contrasts with the control ticks infected with 198 strain only. Other explanations of this interference could include alteration of viral receptors on host cells or induction of antiviral activity in the infected ticks and/or other vectors such as mosquitoes.     

Utility of a Sequence-Independent,Single-Primer-Amplification (SISPA) and Nanopore Sequencing Approach for Detection and Characterization of Tick-Borne Viral Pathogens

Currently, next generation sequencing (NGS) is the mainly used approach for identification and monitorization of viruses with a potential public health threat in clinical and environmental samples. To facilitate detection in NGS, the sequence-independent, single-primer-amplification (SISPA) is an effective tool for enriching virus sequences. We performed a preliminary assessment of SISPA-nanopore sequencing as a potential approach for screening tick-borne viruses in six specimens with detectable Crimean-Congo hemorrhagic fever virus (CCHFV) and Jingmen tick virus (JMTV) sequences. A comparison of unbiased NGS and SISPA followed by nanopore sequencing was carried out in 4 specimens with single and pooled ticks. The approach was further used for genome sequencing in culture-grown viruses. Overall, total/virus-specific read counts were significantly elevated in cell culture supernatants in comparison to single or pooled ticks. Virus genomes could be successfully characterized by SISPA with identities over 99%. Genome coverage varied according to the segment and total read count. Base calling errors were mainly observed in tick specimens and more frequent in lower viral loads. Culture-grown viruses were phylogenetically-related to previously-reported local viruses. In conclusion, the SISPA + nanopore sequencing was successful in generating data comparable to NGS and will provide an effective tool for broad-range virus detection in ticks.     

新疆准噶尔盆地南缘地区新疆出血热病毒自然疫源地的发现

1990年4月下旬我们在准噶尔盆地古尔班通古特沙漠的南缘荒漠地区,从采集的亚洲璃眼蜱(Hyalomm asiaticum asiaticum)中分离到2株可引起乳鼠发病和在细胞培养中复制的病原。经血清学鉴定,包括交叉补体结合,间接免疫荧光和抗新疆出血热McAb(14B_7株)致敏血球的RPHA和RPHI试验证实为新疆出血热病毒。并采集了在荒漠牧场中放牧的羊群血清148份及放牧人员血清21份。用新疆出血热McAb RPHI法从羊血清中查到抗体阳性血清107份(72.3％)人群血清抗体阳性者2份(9.5％)。 上述结果首次证实了新疆北疆的准噶尔盆地南缘荒漠地带存在新疆出血热病毒的自然疫源地,该地区是北疆农牧业生产的主要地区和石油工业的新开发区,也是北疆主要的冬季牧场。