特异性脱氧核酶对小鼠呼吸道合胞病毒感染的抑制作用

目的 研究特异性抑制呼吸道合胞病毒(respiratory syncytial virus,RSV)N基因的脱氧核酶DZ1133在小鼠体内对病毒复制和气道炎症的影响.方法 RSV感染BALB/c小鼠滴鼻给予DZ1133及其变异体、反义寡核苷酸AS1133,空斑形成实验检测肺组织病毒滴度,RT-PCR检测病毒mRNA表达,支气管肺泡灌洗液白细胞计数,ELISA检测TNF-α、IL-12、IFN-γ和IL-10水平,肺组织病理学分析炎症情况.结果 0.2、0.4 mg和0.8 mg DZ1133治疗组小鼠肺组织病毒滴度分别为(2.75±0.21)×105、(1.86±0.20)×105PFU/g和(1.02±0.22)×103PFU/g,与感染对照组小鼠(7.94±0.42)×105PFU/g比较明显下降(P<0.01),上述各剂量治疗组小鼠肺组织病毒mRNA表达与感染对照组比较分别下降30.51%、47.38%(P<0.05)和53.97%(P<0.01).0.4 mg DZ1133治疗降低RSV感染小鼠支气管肺泡灌洗液中白细胞总数,改善肺组织病理学损伤(P<0.05),对TNF-α、IL-12、IFN-γ和IL-10水平无显著性影响(P>0.05).结论 特异性脱氧核酶DZ1133在小鼠体内有效抑制RSV复制、减轻气道炎症,是有效的抗RSV途径.     

反义硫代修饰寡核苷酸体外抗呼吸道合胞病毒感染的研究

目的：在感染呼吸道合胞病毒(RSV)的9HTE细胞中观察与RSVNS1基因和M2基因互补的反义硫代寡核苷酸的抗病毒活性。方法：直接观察RSV的致细胞病变作用，用MTT方法测定细胞存活率。结果：1．RSV感染复数（moi)在0.1以上时，培养5天后RSV感染的9HTE细胞全部死亡。2．我们设计的反义核酸能够提高感染细胞的存活率，且呈量效依赖关系。结论：反义硫代寡核苷酸具有较明显的抗病毒活性，为进一步研究新型抗RSV药物奠定了基础。     

不同年龄患儿对呼吸道合胞病毒感染的免疫反应

目的观察呼吸道合胞病毒（RSV）感染的小于6个月和大干12个月患儿与同年龄对照组相比是否存在年龄相关性的不同细胞因子反应。方法选择60例小于2岁的RSV性毛细支气管炎住院患儿,分为小于6个月和大于12个月两组,并设有同年龄的对照组。通过测定血清细胞因子（IFN-γ、IL-4、IL-10、IL-13）的浓度来观察对RSV感染的免疫反应。结果RSV感染组与对照组相比,IFN-γ、IL-10、IL-13水平在大于12个月年龄组差异有统计学意义（P〈0．05）。而小于6个月年龄组IFN-γ水平差异有统计学意义（P〈0．05）。两组对照组中所有细胞因子均无年龄相关性差异。结论不同年龄婴儿对RSV感染的免疫反应存在差异。     

Prevention of respiratory syncytial virus infection in high risk infants

The incidence of serious RSV illness in premature infants and in infants with CLD can be reduced. Prophylaxis in infants with CHD and cystic fibrosis seem prudent as well, and clinical trials are currently under way to evaluate the use of Synagis in these high-risk groups. Synagis is preferred for most high-risk children because of its ease of administration, safety and effectiveness. The dose for Synagis is 15 mg/kg i.m. monthly during RSV season.     

馥感啉口服液对呼吸道合胞病毒感染小鼠的治疗作用

目的: 研究馥感啉口服液对呼吸道合胞病毒(respiratory syncytial virus, RSV)感染小鼠的治疗作用。方法: 将120只ICR小鼠随机分为对照组,模型组,利巴韦林组,馥感啉5、10和20 g/kg组。采用滴鼻感染法感染RSV后,根据实验分组分别进行灌胃给药治疗,对照组及模型组用等量的生理盐水进行治疗处理,每日2次,共6 d。通过HE染色分析馥感啉口服液对小鼠肺病变的治疗情况,分别采用ELISA和RT PCR方法检测小鼠肺泡灌洗液及肺组织中IFN β、IL 6的含量及mRNA表达水平。流式细胞术检测小鼠外周血CD3+CD4+,CD3+CD8+水平。结果： 成功建立RSV感染小鼠模型。HE病理结果显示不同治疗方法对肺组织病变均有不同情况改善,其中以馥感啉10 g/kg组和20 g/kg组对小鼠肺组织病变改善作用最为明显。馥感啉口服液各剂量组小鼠IFN β和IL 6含量及mRNA表达水平随用药时间延长而逐渐降低;与对照组相比,模型组、利巴韦林组及馥感啉5 g/kg组CD3+CD4+水平显著升高(P均<0.05),馥感啉10 g/kg组和20 g/kg组CD3+CD4+显著降低(P均<0.05);与对照组相比,模型组CD3+CD4+/CD3+CD8+水平显著升高(P<0.05),馥感啉20 g/kg组显著降低(P<0.05),利巴韦林组、馥感啉5 g/kg组及10 g/kg组CD3+CD4+/CD3+CD8+水平与对照组相比,差异均无统计学意义(P>0.05)。结论： 馥感啉口服液各剂量组对小鼠RSV感染有一定的治疗效果,并在一定范围内存在剂量效应关系,高剂量组治疗效果更加显著。     

Mortality in children hospitalised with respiratory syncytial virus infection in Singapore

INTRODUCTIONThis study aimed to investigate the trend and seasonality of infection due to respiratory syncytial virus (RSV) at KK Women’s and Children’s Hospital (KKH) in Singapore and to examine the risk factors for mortality among children with RSV infection requiring admission to the paediatric intensive care unit (PICU).METHODSA retrospective study was conducted at KKH on children with RSV infections who were admitted to the PICU between January 2004 and December 2010. The medical records of children who died from RSV infections were reviewed. Linear regression was performed to determine the risk factors for RSV mortality.RESULTSRSV infection was documented in 5,785 children during the study period; the infection was noted to be occurring throughout the year, with a small increase in prevalence between the months of June and August every year. Among 85 (1.5%) out of 5,785 children who were admitted to the PICU for RSV infection, 74 (1.3%) survived and 11 (0.2%) died. Multivariate logistic regression analysis showed that haemodynamically significant cardiac disease (odds ratio [OR] 12.2, 95% confidence interval [CI] 0.9–16.7, p = 0.05), immunodeficiency (OR 71.4, 95% CI 8.2–500, p < 0.001) and metabolic disease (OR 71.4, 95% CI 4.3–1,000, p = 0.003) were independent risk factors for mortality in RSV infections. Prematurity increased the risk of admission to the PICU but was not significantly associated with mortality.CONCLUSIONChildren with haemodynamically significant cardiac disease, immunodeficiency and metabolic disease were at higher risk of death after hospitalisation for RSV-related illnesses. These children should be considered for palivizumab prophylaxis.     

柴芩感冒药对呼吸道合胞病毒的抑制作用

目的研究柴芩感冒药对呼吸道合胞病毒(RSV)的体外抑制作用.方法微量细胞病变抑制法.从药物的最大无毒量(TD0)开始,作连续2倍系列稀释6个浓度,每一浓度均做4孔,分别加入已感染RSV的单层HeLa细胞中,整个实验重复3次.按Reed-Muench法计算产生50%病变抑制的药物浓度,即药物的半数有效量(IC50),并计算治疗指数(TI).结果柴芩感冒药的IC50为(31±7.07) mg.L-1,TI值为28.88±4.05;中药对照金莲清热颗粒和西药对照利巴韦林注射液的IC50和TI值分别为(36±5.66) mg.L-1、18±2.8和(23.43±5.09)mg.L-1、93±10.7.结论柴芩感冒药的抗RSV效果略好于金莲,但较利巴韦林的药效差.     

空心莲子草有效成分抗呼吸道合胞病毒作用的实验研究

目的 利用呼吸道合胞病毒(RSV)感染的小白鼠模型,观察空心莲子草对小白鼠感染RSV的保护作用.方法 空心莲子草经口服治疗RSV感染的BALB/C鼠,药物剂量为2.5、4.5和6.5 g/(kg·d),疗程5 d,同时以病毒唑作为标准阳性对照,观察动物的发病情况,血液和肺组织内病毒滴度和病毒检出率作为考核药物的疗效指标.结果 空心莲子草4.5g/(kg·d)和6.5 g/(kg·d)剂量作用下的小白鼠其病死率与未经治疗的病毒对照比较,有显著差异;空心莲子草6.5 g/(kg·d)剂量作用后动物血液中病毒检出率为31.3%,肺组织病毒检出率为37.5%,与病毒对照比较有显著性差异,4.5 g/(kg·d)剂量作用后动物肺组织病毒检出率为43.8%,与病毒对照比较有显著性差异;空心莲子草3个不同剂量作用下小鼠的肺指数与病毒对照比较均有显著性差异.结论 空心莲子草有望成为治疗RSV感染的有效药物.这亦为中草药抗病毒作用的研究开辟了新的途径.     

Molecular epidemiology of respiratory syncytial virus

Objective To determine the epidemiologic pattem of subgroups A and B and genotypes of respiratory syncytial virus (RSV) during two noncontinuous epidemics during 1990- 1991 and 1997- 1998 in Beijing. Methods Nasopharyngeal secretion (NPS) samples of RSV positive or RSV isolates tested by indirect immunofluorescence (IIF) assay were classified into subgroups A and B. Isolates of RSV were divided into at least six different lineages, designated NP1-NP6, by restriction mapping of the N gene. Np1, 3 and 6 were given by subgroup B isolates, while NP2, 4 and 5 were given by subgroup A isolates. Strains of subgroup A were further subdivided into six lineages SHL1-SHL6 on the basis of the SH gene sequence. SH lineages were closely related to each other and to NP1-NP6. Strains of SHL1, 3 and 4 were closely related and belonged to NP2, SHL2 and 6 to NP4, and SHL5 to NP5. Results Of 145 RSV NPS samples from the 1997- 1998 epidemic, 83 (57.2％) were of subgroup B RSV positive, 62 (42.8％) of subgroup A RSV positive. The rate of occurrence of subgroup A to B strains was about 1:1.3. Two of 10 isolates during the epidemic were subgroup A strains, whereas 8 were subgroup B strains. The rate of occurrence of subgroup A to B strains was 1:4. Eight subgroup A strains of 10 isolates from the 1990 - 1991 epidemic were dominant; the proportion of subgroup A to B strains was 4:1. With 10 RSV isolates in 1997- 1998, all 2 subgroup A strains gave N gene fragment restriction pattern NP4, and fell into SH lineage SHL2, whereas 8 subgroup B strains all belonged to NP3. All 8 subgroup A isolates from the 1990- 1991 epidemic gave pattem NP4, and fell into SHL2, while 2 subgroup B strains all belonged to NP3. The classification of subgroups A and B deduced from NP pattems corresponded to the definition of these subgroups by monoclonal antibodies. Conclusions These observations confirm that subgroups A and B or multiple lineages of RSV co circulated in Beijing, but different genome types predominated each year. Moreover, very similar viruses were isolated up to more than 5 years ago, indicating that despite apparent diversity of the subgroup A strains, the separate lineages might be relatively stable.     

Toll3受体在呼吸道合胞病毒感染中作用的实验研究

目的探讨呼吸道合胞病毒（RSV）感染人肺上皮A549细胞后,Toll样受体3（TLR3）的水平变化及其产生的I型干扰素的抗病毒作用。方法RSV感染体外培养的人肺上皮A549细胞,并给予TLR3特异性抗体处理,分别感染4h、8h、12h、16h和24h后收集各组细胞。未感染病毒的细胞作为对照组。lit—PCR法检测TLR3、IFN-α、IFN—β,RSVF蛋白的mRNA表达水平变化。结果RSV感染A549细胞后,TLR3、IFN—α、IFN-β,RSVF蛋白的mRNA表达量均升高且有时间依赖性。结论RSV感染A549细胞后可上调TLR3表达,其活化细胞介导产生的I型干扰素能起到抗病毒作用。     

反转录—聚合酶链反应在呼吸道合胞病毒感染检测的应用

为呼吸道合胞病毒（ＲＳＶ）感染的临床诊断寻求敏感，特异的检测方法，采用ＲＴ－ＰＣＲ技术扩增毒培养液和急性呼吸道感染患者鼻咽分泌物中的ＲＳＶ基因，并应用地高辛标记ｃＤＮＡ探针对扩增产物进行鉴定。结果成功地提取了ＲＳＶｍＲＮＡ，反转录合成ｃＤＮＡ，经扩增得到４７１ｂｐ左右ｃＤＮＡ区带。流感病毒Ｂ，副流感病毒２型，腺病毒７对照无扩增带，ＲＳＶ培养液１０倍连续稀释至１：１０００，经ＲＴ－ＰＣＲ扩增仍可见４     

一种快速诊断呼吸道合胞病毒感染的方法

应用引物设计原则，选择呼吸道合胞病毒（ＲＳＶ）基因组中高度保守的Ｆ基因作为扩增靶序列，独立设计合成了两对引物，建立了一个逆转录套式聚合酶链反应（ＰＣＲ）检测ＲＳＶＲＮＡ的方法。结果：对引物进行敏感度实验，其检测敏感度为每ｍｌ１０ＴＣＩＤ５０ｓ（５０％ｔｉｓｓｕｅｃｕｌｔｕｒｅｉｎｆｅｃｔｉｖｅｄｏｓｅ）的病毒量；用副流感病毒１型、甲型流感病毒、乙型流感病毒、新城疫病毒、腮腺炎病毒和腺病毒２型作对照，均无预期扩增产物出现，证实了设计引物的特异性；用建立的逆转套式ＰＣＲ方法对５例急性下呼吸道感染患儿的鼻咽部分泌物标本进行检测，发现３例阳性，２例阴性；同时用病毒培养法检测，结果完全相同。提示：该方法具有快速、敏感、特异和观察结果客观等优点。     

呼吸道合胞病毒感染对鼻黏膜下成纤维细胞分泌趋化因子的影响

[目的]了解呼吸道合胞病毒（RsV）感染对鼻黏膜下成纤维细胞分泌趋化因子的影响．[方法]取手术切除的下鼻甲鼻黏膜进行上皮细胞及黏膜下成纤维细胞培养,将RSV感染鼻黏膜上皮细胞后,用其培养上清液刺激培养的成纤维细胞,用荧光定量PCR分别于6,12,24,48h时测定嗜酸细胞活化趋化因子、调节活化正常T细胞表达和分泌的趋化因子（RANTES）、白细胞介素8（IL－8）及生长调节癌基因α（GRO—α）的mRNA表达程度;用ELISA方法分别于24,48h测定培养液中嗜酸细胞活化趋化因子、RANTES,IL-8,GRO-α蛋白质的含量．[结果]实验组IL-8,GRO-αmRNA的表达程度明显高于对照组,RANTES,IL-8,GRO-α蛋白表达量明显高于对照组．[结论]被RSV感染的鼻黏膜组织中成纤维细胞有     

ARDS in an HIV-positive patient associated to respiratory syncytial virus

We describe a clinical case of ARDS in an HIV infected patient. ARDS was associated to a respiratory syncytial virus infection that triggered a suspected Pneumocystis infection that despite missing etiologic proofs was treated with antimycotics. As rather limited information on RSV associated ARDS in HIV patients is available in the current literature, this case is of significant interest.     

呼吸道合胞病毒感染对肺组织GRK2的影响及其与哮喘的关系

目的 :探讨呼吸道合胞病毒(respiratory syncytial virus,RSV)感染鼠肺组织后,G蛋白偶联受体激酶2(G protein coupled receptor kinase 2,GRK2)的变化规律及其与哮喘的关系。方法:Balb/c雌性小鼠100只,随机分为5组,每组20只。正常对照组,于第1、14天予生理盐水0.2 ml腹腔注射,第21~25天用37℃生理盐水20 ml雾化吸入30 min;RSV感染组,于第19、20、21天,以RSV按浓度1×106PFU、100μl/次鼻腔滴入;哮喘组,第1、14天予以鸡卵白蛋白(ovalbumin,OVA)100μg、AL(OH)32 mg腹腔注射,第21天开始激发,以2%OVA生理盐水20 ml雾化吸入30 min,持续激发5 d;双重感染组,模型制备同哮喘组,第19、20、21天,以RSV按浓度1×106 PFU、100μl/次鼻腔滴入;地塞米松干预组,RSV感染同RSV组,第21~25天予腹腔注入地塞米松0.2 mg/(kg·d)。末次激发后24 h,10%水合氯醛腹腔注射麻醉(0.3 ml/kg),处死小鼠,取左肺上叶和右肺中叶,4%甲醛固定,分别用于免疫荧光检测GRK2的表达和组织病理改变检测;取右肺下叶用于Western blot检测各组GRK2的表达;无菌条件下取左肺下叶,用于肺组织病毒分离及免疫荧光鉴定。结果:RSV组GRK2表达较正常组显著增加,与哮喘组相比差异有统计学意义;双重感染组GRK2表达较RSV组显著增加;地塞米松干预组GRK2表达较RSV组显著减少。结论:RSV感染小鼠肺组织后GRK2表达增加,合并哮喘时表达显著增加,地塞米松干预对GRK2的表达有一定的抑制作用。