麻疹病人咽拭子标本的麻疹病毒分离结果

目的收集麻疹病人的咽拭子标本进行麻疹病毒分离,为我国麻疹病毒流行毒株基因型的鉴定提供更多的分离株。方法2005年5￣8月,选择南京市第二医院收治入院的疑似麻疹病人109例采集了咽拭子标本,接种EB病毒转化的狨猴淋巴母细胞(B95a细胞)。结果分离到麻疹病毒46株,分离阳性率为42.20%。结论此次南京第二医院与江苏省CDC合作分离到麻疹野病毒,标本量大,阳性率高,分析原因,与采集的时间、操作人员的技术及冷链储运和及时运输密切相关。     

Oncolytic measles viruses for cancer therapy

New strategies using biological agents are being developed to treat cancer. Live viruses are among these new agents. Virotherapy uses replication-competent viral vectors with strong oncolytic properties. With the use of molecular virology techniques, viruses have been genetically engineered to replicate selectively in tumour cells and are under preclinical and clinical investigation at present. Measles virus (MV) is being used for this purpose. Replication-competent attenuated Edmonston B measles vaccine strain (MV-Edm) is non-pathogenic and has potent antitumour activity against several human tumours. The virus is selectively oncolytic in tumour cells, eliciting extensive cell-to-cell fusion and ultimately leading to cell death. Therefore, MV-Edm is a safe and efficient means to kill tumour cells. Further improvements in existing MV vectors may increase tumour selectivity and oncolytic activity. This review discusses the discovery and development of replication-competent oncolytic MV for cancer therapy.     

广东省东莞市1株麻疹野病毒核蛋白基因分析

目的分析2008年广东省东莞市疫情中分离到的1株麻疹病毒的核蛋白基因序列,了解毒株基因型别及其核蛋白基因变异情况。方法采集麻疹疫情中病例的尿液标本,用VERO-SLAM细胞进行病毒分离培养,对分离到的病毒株通过反转录-聚合酶链反应（RT-PCR）扩增麻疹病毒核蛋白基因片段,对扩增产物进行测序并与麻疹各基因型代表株及沪191疫苗株进行序列比对和构建进化树分析。结果分离到的麻疹病毒株属于H1基因型H1a亚型。与China93-2毒株同源性最高,为99.1%,与China94-7毒株同源性为97.1%,与其他几株H1亚型的同源性则在97.6%～98.0%之间。与沪191疫苗株同源性为92.9%。结论此次疫情中的毒株属于H1基因型H1a亚型,与中国其他地区的流行情况一致。与其同亚型的毒株相比变异较小。     

浙江地区2012年麻疹病毒流行株基因特征研究

摘要:目的：探讨浙江地区2012年3～9月麻疹病毒（MV）的基因特征。 方法：用逆转录-聚合酶链反应（RT-PCR）检测麻疹患者的咽拭子标本,对部分PCR扩增阳性的片段进行基因测序并绘制系统进化树进行同源性分析。 结果：88例麻疹患者咽拭子标本中经RT-PCR检测阳性76例,阴性12例;随机选择13例阳性标本进行基因测序,结果显示均为H1a亚型;系统进化树分析结果表明,13株均为H1a基因亚型,与该型参考株AF045205、AF045206核苷酸同源性为98.0%～99.0%。 结论：H1a基因型是浙江地区2012年MV流行的优势亚型,无输入性的其他亚型出现。     

孕妇静脉血麻疹IgG抗体水平的影响因素研究

目的研究不同年龄、麻疹病史及麻疹疫苗接种史对孕妇静脉血麻疹病毒(MV)IgG抗体水平的影响,为预测母传抗体对其婴幼儿的保护作用提供科学的理论依据。方法选取2013年2月至2015年2月来广东省深圳市福田区妇幼保健院产科分娩,且符合纳入排除标准的217例孕妇作为研究对象,采用酶联免疫吸附试验(ELISA)定量检测孕妇静脉血中MV IgG抗体水平。结果不同年龄组之间的麻疹IgG抗体几何平均浓度(GMC)均高于阳性值同时均低于有效保护值,且差异均无统计学意义(P0.05);不同年龄组之间的MV IgG抗体的阳性率与保护率经比较差异均无统计学意义(P0.05)。有MV疫苗接种史的孕妇其血清MV IgG抗体的GMC明显低于有MV病史的孕妇其血清MV IgG抗体的GMC,且两组GMC差异有统计学意义(P0.05);MV疫苗接种史组的保护率(37.9%)明显低于MV病史组的保护率(85.7%),且差异具有统计学意义(P0.05);其余指标检测差异均无统计学意义(P0.05)。结论年龄对孕妇体内MV IgG抗体无显著影响;不论是具有麻疹病史或麻疹疫苗接种史,其孕妇静脉血麻疹IgG抗体GMC对婴儿的保护率低,建议育龄妇女孕前再次接种麻疹疫苗。     

Inter‐operator variation in ELISPOT analysis of measles virus‐specific IFN‐γ‐secreting T cells

The ELISPOT assay is a highly sensitive technique used for the detection of individual cytokine releasing cells. We have developed an IFN‐γ ELISPOT assay utilizing unfractionated frozen peripheral blood mononuclear cells (PBMC) to quantify the frequency of measles virus (MV)‐specific IFN‐γ‐secreting T cells in 117 healthy children who had been previously immunized with two doses of the measles‐mumps‐rubella vaccine. We have also estimated the variability associated with the quantification of ELISPOT plates and compared the number of MV‐specific IFN‐γ‐secreting T cells for each subject as determined by two different operators of an ELISPOT reader. The median frequency of MV‐specific IFN‐γ‐producing memory T cells detected by this assay was 0.005 % and 0.01 % as determined by an in‐house and commercial operator, respectively. Although we found a significant correlation (r = 0.83, p<0.0001) between the number of spots counted by the commercial and in‐house operators of an ELISPOT reader, the median number of spots counted by the commercial operator was twice the number of spots counted by an in‐house operator (p<0.001). This demonstrates the importance of using a common ELISPOT reader and operator, among other parameters, to quantify the number of spots when a large volume of plates are being scanned and analyzed.     

Measles virus induces apoptosis in uninfected bystander T cells and leads to granzyme B and caspase activation in peripheral blood mononuclear cell cultures

Background Measles causes lymphopenia and depresses cell‐mediated immunity, but the mechanisms of immunosuppression and cell loss are poorly known. Methods We have used an in vitro model of measles virus (MV)‐infected peripheral blood mononuclear cells (PBMCs) and phytohaemagglutinin‐stimulated PBMCs in order to assess MV–leucocyte interactions. Cell population undergoing apoptosis was measured by flow cytometry and Annexin‐V‐fluos staining. The expression of Fas, FasL, TNRF1, and Bcl‐2 was analyzed by flow cytometry and Western blotting, and activation of caspase cascade was measured using a colourimetric caspase substrate set. The effects of caspase inhibitors were detected by flow cytometry. Results Measles virus was able to infect monocytes, but interestingly induced apoptosis in uninfected T cells, indicating that induction of apoptosis in T cells is mediated by MV‐infected adherent cells. Only 1% of T cells contained MV antigen day 3 p.i. Interestingly the percentage of early apoptotic T cells at the same time was 35%, showing that apoptosis was not the result of MV infection in T cells. Measles virus‐induced Fas but not FasL or TNFR1 expression on PMBC, as well as activation of granzyme B and caspase cascade. Simultaneously, overexpression of Bcl‐2 protein was detected. Caspase inhibitor decreased the amount of apoptotic T cells. Conclusion Measles virus‐infected monocytes induce apoptosis in uninfected T cells, suggesting that infected monocytes probably interact via cell–surface molecules with uninfected T cells and induce apoptosis by indirect mechanisms. Apoptosis of the lymphocytes may contribute to the pathogenesis of MV‐induced immunosuppression and cell loss.     

麻疹病毒多肽抗原免疫原性分析

目的分析麻疹病毒(measles virus,MV)主要多肽抗原成分,筛选可用于麻疹病毒特异性抗体诊断的抗原成分。方法应用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹技术(immunoblots,IB)对比分析细胞抗原与麻疹病毒抗原,确定病毒多肽抗原的存在,利用不同时期免疫动物的多克隆抗体分析病毒多肽抗原免疫原性的强弱。结果SDS-PAGE和多克隆抗体免疫印迹结果表明,麻疹病毒分子量为60 kD和55 kD抗原的免疫原性最强,72 kD抗原性较弱,200 kD抗原性最弱。结论针对麻疹病毒60 kD和55 kD抗原成分的抗体检测可以作为麻疹病毒感染的早期诊断指标。     

2013年北京市首例输入性D_9基因型麻疹毒株的分离和鉴定

目的分离并鉴定北京市输入性D9基因型麻疹毒株。方法使用Vero/SLAM细胞,对可疑麻疹输入性病例的咽拭子和尿液标本进行麻疹毒株的分离培养。用反转录-聚合酶链反应扩增麻疹病毒核蛋白(N)基因羧基末端676个核苷酸片段,对扩增产物进行核苷酸序列测定和分析,并以羧基末端450个核苷酸片段构建基因亲缘关系树,进行遗传距离及核苷酸同源性分析。结果该病毒分离株BJCY13026-2和世界卫生组织D9基因型代表株Victoria.AUS(维多利亚.澳大利亚)12.99在基因亲缘性关系树上同属一个分支,核苷酸同源性为95.8%,氨基酸同源性为96%;和其他23个基因型代表株的核苷酸和氨基酸同源性分别在88.1%~95.6%和90.7%~96.7%。和中国大陆目前所使用的麻疹疫苗株沪191相比对,其核苷酸和氨基酸同源性分别为91.1%和90.0%;和中国目前流行的麻疹病毒绝对优势本土基因型H1a基因型代表株相比对,其核苷酸和氨基酸同源性分别为89.4%和91.3%。结论该输入性病例的病毒分离株为麻疹病毒D9基因型。     

1957-2006年浙江省奉化市麻疹流行病学分析

目的 分析奉化市1957-2006年50年麻疹流行病学特征,为调整麻疹免疫策略提供依据.方法 将50年麻疹疫情分为麻疹疫苗接种前、麻疹疫苗推广应用、麻疹疫苗纳入计划免疫一年一次突击接种和麻疹疫苗冷链装备按月接种四个阶段.前三个阶段年均发病率用几何均数,第Ⅳ阶段用算术均数.结果 四个阶段麻疹年发病率和年龄构成有显著差异;发病年龄构成由幼儿和学龄儿童为主向8月龄小婴儿和15岁以上人群转移,2005-2006年奉化市本地人口8月龄小婴儿和15岁以上人群发病占总病例数的96.92%.结论 为降低大小两端年龄麻疹发病,达到消除麻疹的目标,建议对初中学生加强接种麻疹疫苗.     

Measles—What’s Old Is New Again

Measles (rubeola) was once nearly eradicated in the United States. Unfortunately, it has reappeared, with more than three times the number of confirmed cases in 2019 than in 2018. The virus, which produces a distinct rash that appears within days of exposure, can spread quickly and can produce severe complications. There is no cure; treatment is supportive care. Measles was once a minimal concern in the United States due to high vaccination rates. The Centers for Disease Control and Prevention reports that, because of the increase in individuals traveling to countries with high rates of measles and the fact that fewer U.S. children are being immunized against measles, the incidence of measles will likely continue to rise in the United States. It is vital that nurses and other health care providers educate individuals about the importance of the prevention and treatment of measles.     

Oncolytic measles virus strains as novel anticancer agents

Introduction: Replication-competent oncolytic measles virus (MV) strains preferentially infect and destroy a wide variety of cancer tissues. Clinical translation of engineered attenuated MV vaccine derivatives is demonstrating the therapeutic potential and negligible pathogenicity of these strains in humans.

Areas covered: The present review summarizes the mechanisms of MV tumor selectivity and cytopathic activity as well as the current data on the oncolytic efficacy and preclinical testing of MV strains. Investigational strategies to reprogram MV selectivity, escape antiviral immunity and modulate the immune system to enhance viral delivery and tumor oncolysis are also discussed.

Expert opinion: Clinical viral kinetic data derived from noninvasive monitoring of reporter transgene expression will guide future protocols to enhance oncolytic MV efficacy. Anti-measles immunity is a major challenge of measles-based therapeutics and various strategies are being investigated to modulate immunity. These include the combination of MV therapy with immunosuppressive drugs, such as cyclophosphamide, the use of cell carriers and the introduction of immunomodulatory transgenes and wild-type virulence genes. Available MV retargeting technologies can address safety considerations that may arise as more potent oncolytic MV vectors are being developed.     

1950～2004年乐山市麻疹流行特征与防制对策探讨

目的了解乐山市麻疹流行特征与规律,探讨防制对策与措施。方法采用一般统计方法和卡方检验方法,用SPSS10.0软件处理。结果开展计划免疫后乐山市发病率下降了92.15%、死亡率下降了98.85%,发病年龄呈后移趋势(!2=283.26,P<0.0001),发病职业趋向学生(!2=359.24,P<0.0001),少、边地区是乐山市计划免疫的薄弱环节。结论开展10岁以上在校生和新入学大、中专生麻苗复种,加强麻疹流行病学和实验室监测以及少、边地区常规免疫接种质量的督导检查和完善冷链设备,是防止麻疹在乐山市暴发流行的重要策略。     

成人麻疹患者的护理

笔总结27例成人麻疹患的护理经验；严格执行呼吸道隔离，防止疾病传播，及时补充水分及营养，促进早日康复，加强全身护理，防止并发症；密切观察病情变化，做好高热的处理，加强并发症护理，做好心理护理，消除患的顾虑。     

浙江省某沿海城市麻疹疫情现场流行病学调查

目的 调查和分析浙江省某市麻疹疫情流行特征,查明麻疹疫情流行的原因,探讨麻疹防治对策.方法 现场流行病学调查,描述性流行病学分析,用血凝抑制(HI)法检测麻疹IgG抗体.结果 2004年1月至2005年2月28日,全市共报告麻疹病例599例,男性343例,女性256例.2004年11月麻疹疫情开始流行,12月至次年1月达到高峰,病例分布广泛,散发和局部暴发并存.病例中无免疫史的占62.02%,免疫史不详占22.11%.共采集育龄妇女血清样本109份,其中抗体阳性105份,阴性4份,阳性率为96.33%,几何平均滴度为1:8.0.结论 需要加强麻疹疫苗的常规免疫工作,同时要考虑对大年龄组人群接种麻疹疫苗,继续加强麻疹监测,探讨流动儿童的计划免疫管理模式.     

麻疹合并肺炎患儿的临床观察与护理

目的总结麻疹合并肺炎患儿的临床表现及相关护理方法,以期为基层医院的临床护理工作提供经验。方法回顾性分析2005-2010年在温州医学院附属第二医院确诊为麻疹合并肺炎患儿的临床资料,统计患儿的一般情况、临床特点,分析并总结其预防及护理措施。结果所有患儿均有发热、咳嗽和全身斑丘疹等症状。体温37.8～41.0℃,热程3～12d,平均(7.8±2.3)d;典型皮疹467例,占90.2%,出疹期平均(3.5±1.7)d。肺部可闻及细小湿啰音261例,占50.4%;可闻及中等湿啰音187例,占36.1%;可闻及哮鸣音163例,占31.5%;未闻及明显干湿性啰音87例,占16.8%。肺部体征平均在热退后(3.8±1.2)d好转。518例患儿中,513例(99.0%)患儿治愈或好转出院,2例自动出院,3例死亡。结论科学全面的护理干预措施可有效促进麻疹合并肺炎患儿的康复。     

175例麻疹流行病学调查分析

目的了解麻疹流行病学特征。方法采用回顾性调查方法对2005年1￣6月湖州市中心医院收治的175例麻疹病例资料进行分析。结果麻疹抗体总阳性率为55.4%(97/175)。麻疹流行特点是散发和小范围流行同时存在。发病有明显的季节性,1月份发病开始上升,5月份下降。发病以青壮年为主,占58.6%,其次为10岁以下儿童,占20.0%。男女性别比为1∶1.3。结论除儿童外,青壮年也为易感人群,应采取相应措施重点预防和保护。     

2000～2004年杭州市萧山区麻疹疫情流行病学分析

目的了解近几年杭州市萧山区麻疹流行病学特征,加速控制麻疹。方法对2000￣2004年麻疹临床和血清学诊断病例进行流行病学分析。结果萧山区麻疹疫情以散发为主,存在个别暴发点,流动人口比率呈明显上升趋势。发病高发季节为2￣6月。本地病例以<1岁婴幼儿为主,5￣9月龄病例占1岁以内病例的92.31%;流动人口以1岁以上低年龄儿童及学龄儿童为主。15岁以下(不包括不足8月龄婴幼儿)本地和流动人口病例中无免疫史者分别为39.19%、65.12%,本地病例明显低于流动人口。结论继续加强常规免疫接种工作,保持高水平的实际免疫覆盖率和免疫成功率;加强流动儿童的免疫接种管理,消除流动人口的免疫空白,是萧山区控制和消除麻疹的重点。