清肺口服液含药血清对呼吸道合胞病毒抑制作用的实验研究

目的研究清肺口服液含药血清对呼吸道合胞病毒(RSV)感染细胞的抑制作用。方法用不同剂量的清肺口服液制成含药血清处理呼吸道合胞病毒感染后的Hep-2细胞,用组织细胞培养法,采用细胞病变(CPE)观察和四氮唑盐还原法(MTT法)检测清肺口服液含药血清对RSV感染细胞的增殖抑制作用。结果在Hep-2细胞上,清肺口服液的最大无毒浓度为1∶9。100TCID50RSV攻击后Hep-2细胞的OD值明显降低,加入中高剂量含药血清后OD值升高。结论清肺口服液含药血清对RSV有抑制作用。      

热毒宁抗呼吸道合胞病毒(RSV,Long株)作用体外实验研究

目的:研究中草药热毒宁注射液对呼吸道合胞病毒(RSV,Long株)的抑制作用.方法:以病毒唑为阳性对照药,采用细胞培养技术,观察不同药物浓度及不同给药方式下RSV攻击后各组Hep-2细胞的病变效应(CPE),在此基础上采用MTT比色法,测定各组细胞的病毒抑制率.以CPE法计算药物的半数中毒浓度TC50,分别以CPE法及MTT法计算药物的半数有效浓度EC50及治疗指数TI,比较不同药物浓度及不同给药方式下热毒宁对呼吸道合胞病毒(RSV)的抑制作用效果.结果:热毒宁注射液半数中毒浓度TC50为3.972g/L.预防给药方式、细胞外直接灭活及治疗给药方式均有抗RSV作用,其抗RSV的半数有效浓度(EC50)分别为0.428、1.160、1.189 g/L,治疗指数TI分别为9.28、3.42和 3.34,热毒宁对RSV的抑制作用存在着明显的量效反应关系.结论:热毒宁对RSV有直接灭活作用,对RSV侵入Hep-2细胞有阻断作用,对RSV在Hep-2细胞内增殖有抑制作用,相同浓度下,以预防作用更显著.     

鱼腥草注射液治疗呼吸道合胞病毒感染的体外实验研究

目的探讨鱼腥草注射液体外抗呼吸道合胞病毒(RSV)的有效性。方法采用MTT比色法检测鱼腥草对Hep2单层细胞的毒性作用;以利巴韦林为对照,通过空斑试验和试管观察细胞病变法确定鱼腥草对RSV的抑制作用。结果鱼腥草最大稀释度(1∶10)对细胞无毒性,不影响细胞形态及生长;鱼腥草(1∶10)和利巴韦林(5 mg/L)均可以完全抑制稀释度为10-5(100TCID50)的RSV、半数抑制10-4(1 000 TCID50)的RSV,二者均可使空斑明显变小。结论鱼腥草注射液对RSV有体外抑制作用,对培养细胞无毒性。     

穿心莲提取物体外抗呼吸道合胞病毒作用的研究

目的：了解穿心莲提取物对呼吸道合胞病毒(RSV)的体外抑制作用。方法：通过光镜观察不同药物浓度下：RSV对Hep-2的致病作用(CPE),四甲基偶氮唑盐(MTT)比色检测穿心莲提取物对RSV的抑制和对：RSV感染HeF2细胞的保护作用。结果：穿心莲提取物对RSV无直接灭活作用,亦不能阻止RSV的吸附,各药物浓度均出现典型的CPE;穿心莲提取物有抑制RSV生物合成的作用,其半数抑制浓度(IC50)是1．52μg,／ml,治疗指数为36,且随着药物浓度的增加,CPE特征明显逐渐减弱,病毒滴度逐渐降低,而病毒抑制率则明显增高。结论：穿心莲提取物在体外可能是通过抑制生物合成来发挥抗RSV作用。     

鸡骨草冷浸提取物抗病毒活性研究

目的 研究中药鸡骨草冷浸提取物的抗病毒活性。方法 采用细胞体外抗病毒实验法,结合CPE法及CCK-8试剂盒,以TI值（治疗指数）为研究指标,以利巴韦林、阿昔洛韦为阳性对照药,研究鸡骨草冷浸提取物对呼吸道合胞病毒（RSV）、单纯疱疹病毒（HSV-1）、柯萨奇病毒（COX-B5）和手足口病毒（EV71）的抑制作用。结果 甲醇冷浸提取物抗病毒效果较好,对COX-B5的TI值为16.476,最大抑制率为88%,与阳性对照药利巴韦林（TI值为17.482,最大抑制率为85%）相差不大。结论 鸡骨草冷浸提取物具有体外抗RSV、HSV-1、COX-B5的活性。     

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.     

金欣口服液含药血清抗呼吸道合胞病毒作用研究

目的：观察金欣口服液对呼吸道合胞病毒（RSV）感染的人喉癌上皮细胞（Hep-2）细胞的抑制作用。方法：通过体外抗病毒实验采用MTT法检测病变抑制率及以上清滴度观察金欣口服液含药血清对RSV的抑制作用。结果：MTT法测定病毒抑制率均高于75％,含药血清组上清滴度与空白血清组和病毒对照组比较有显著差异,滴度低于后两组。结论：金欣口服液对呼吸道合胞病毒引起的细胞病变有明显抑制作用。     

荔枝核黄酮类化合物体外抗呼吸道合胞病毒的作用

目的: 探讨荔枝核黄酮类化合物体外抗呼吸道合胞病毒(RSV)的作用. 方法: 采用细胞培养技术,以病毒唑为阳性对照,观察比较荔枝核黄酮类化合物对RSV引起的细胞病变(CPE)的抑制作用. 结果: 荔枝核黄酮类化合物半数中毒浓度(TC50)为152.9 mg/L,其抗RSV的半数有效浓度(IC50)为58.6 mg/L,治疗指数(TI)为2.6,体外抗RSV作用与病毒唑相当,且对RSV抑制作用存在明显的量效关系. 结论: 荔枝核黄酮类化合物在Hep-2细胞中对呼吸道合胞病毒有抑制作用.     

大黄虫丸含药血清对人肺腺癌细胞A549增殖的影响

目的:观察大黄虫丸含药血清对人肺腺癌细胞A549增殖的影响。方法:12只SD大鼠随机分为大黄虫丸组(DHZCW组)、鸦胆子组、生理盐水组,按常规方法制备含药血清。然后以细胞培养液将含药血清稀释成不同的浓度(40%、20%、10%、5%),采用MTT法检测不同浓度含药血清对A549细胞增殖的影响。结果:与生理盐水组比较,20%、40%DHZCW组和20%、40%鸦胆子组OD值降低,差异有统计学意义(P<0.05);与鸦胆子组比较,40%DHZCW组OD值降低,差异有统计学意义(P<0.05)。结论:大黄虫丸含药血清与鸦胆子油口服乳液含药血清均具有体外抑制人肺腺癌A549细胞增殖的作用。     

Effect of Serum Containing Tougu Xiaotong Capsule on Activity of Chondrocyte

目的 观察透骨消痛胶囊含药血清对软骨细胞活性的影响. 方法 SD大鼠膝关节软骨建立软骨细胞体外培养体系,第3代软骨细胞同步化后进行干预,分别确定含药血清的有效干预时间及干预浓度.各组合药血清在有效浓度作用下干预软骨细胞,在有效干预时间点采用MTT法. 结果 干预后,软骨细胞OD值干预72 h明显高于干预24 h、48 h、96 h(P＜0.05);10％含药血清浓度组明显高于5％、20％含药血清浓度组(P＜0.01).确定72 h为有效作用时间,10％为有效作用浓度.在10％含药血清浓度的作用下干预72 h,第3代软骨细胞OD值,透骨消痛胶囊实验组明显高于空白组(P＜0.01),实验2组增殖效果最明显(P＜0.05). 结论 透骨消痛胶囊含药血清能加速软骨细胞增殖,维持软骨细胞活性,其促进软骨细胞增殖作用与药物浓度有关,以中剂量药物浓度(含透骨消痛胶囊29 mg/d)为最佳.     

重组人干扰素α-2b体外抗呼吸道合胞病毒作用的实验研究

目的观察重组人干扰素α-2b在体外的抗呼吸道合胞病毒作用。方法观察不同浓度重组人干扰素α-2b抑制呼吸道合胞病毒对Hela细胞产生细胞病变（CPE）的作用,并与利巴韦林比较。结果实验表明重组人干扰素α-2b抗呼吸道合胞病毒的最低有效浓度为25IU／ml。结论重组人干扰素α-2b具有明显的抗呼吸道合胞病毒作用。     

Detection of respiratory syncytial virus in nasopharyngeal secretions by 24-well plate precentrifugation assay using a monoclonal antibody against F protein

BACKGROUND: Respiratory syncytial virus (RSV) is responsible for 50% of all bronchiolitis and 25% of pneumonia cases during the first month of life. Detection of the RSV antigen by immunofluorescence in exfoliated nasal epithelium or by other methods in nasopharyngeal swabs is useful in the potentially infected patient because results are available within a few hours. In contrast, RSV antigen detection in cell culture may require as much as 3 weeks. METHODS: Three methods for detection of respiratory syncytial virus in 131 clinical respiratory specimens from patients with acute respiratory disease and bronchiolitis were compared utilizing the following: a precentrifugation immunofluorescence assay using Hep-2 cells, indirect immunofluorescence assay, and conventional tube cell culture using Hep-2 cells. RESULTS: Respiratory syncytial virus was identified in 36 specimens by the three methods previously described. The virus was recovered in 41 (31.3%) samples by precentrifugation immunofluorescence assay, 40 (30.5%) were identified by the immunofluorescence technique, and 38 (29.0%) cases were positive by conventional cell culture. The sensitivity of the precentrifugation assay in relation to the immunofluorescence technique was 90%, the specificity 94.5%, and the agreement, 96.2%. A positive predictive value of 90.2% was obtained. Sensitivity, specificity, agreement, and positive predictive values obtained by the precentrifugation assay variant compared to the conventional cell were 90.8%, 94.5%, 93.1%, and 87.8%, respectively. CONCLUSIONS: The precentrifugation immunofluorescence assay method was as sensitive as the remainder of the methods used in our study and represents a valid alternative for rapid detection of respiratory syncytial virus in clinical samples.     

Ribavirin treatment of experimental respiratory syncytial viral infection. A controlled double-blind study in young adults

The effect of ribavirin aerosol on experimental respiratory syncytial virus (RSV) infection was evaluated in a double-blind controlled study of 16 young adult volunteers. Two days after intranasal inoculation with RSV, half of the subjects were treated with a small-particle aerosol of ribavirin and half with placebo for a total of 12 hours each day for three days. Seven of eight placebo-treated and six of eight ribavirin-treated subjects became infected. Viral shedding was diminished in the ribavirin-treated group. The proportion of infected volunteers still shedding virus on days 6 through 9 was significantly less than in the placebo-treated group. Ribavirin appeared to have no effect on minor upper respiratory tract signs, but systemic complaints and fever occurred significantly less often in the ribavirin-treated group. Ribavirin aerosol therapy was well tolerated and produced no significant changes in pulmonary function test results or signs of toxicity. This suggests that such therapy might be further evaluated in infants with RSV lower respiratory tract disease.     

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。     

Refractory congestive heart failure after ribavirin in infants with heart disease and respiratory syncytial virus

Although treatment with ribavirin has been known to be associated with a decreased mortality in infants with congenital heart disease (CHD) who have respiratory syncytial virus (RSV), few data are available regarding morbidity. We reviewed records of 10 consecutively hospitalized infants with CHD during a recent RSV epidemic. Despite the presence of left-to-right shunt in each patient, symptoms of RSV were respiratory at presentation. After ribavirin, decreased respiratory symptoms were found in 8 infants but in 2 assisted ventilation were required 1 and 3 days after admission. Congestive heart failure worsened in 8 patients, 6 of whom had improved respiratory status after ribavirin. Of the 8 patients with worse CHF (pulmonary edema), 3 responded to medical management but 5 were refractory and 4 required surgical repair of CHD. One patient died of pulmonary hemorrhage. Medically refractory CHF may develop in infants with CHD who become infected with RSV and are treated with ribavirin. Further studies are needed to determine whether the pulmonary edema is caused by RSV, ribavirin, or combination of effects on pulmonary capillary function or some other unrecognized mechanism.     

盐酸阿比朵尔抗呼吸道合胞病毒机制初探

目的:明确盐酸阿比朵尔抗吸道合胞病毒(RSV)Long株的活性并探讨其抗病毒机制。方法:通过细胞病变效应(CPE)、噻唑蓝(MTT)比色法检测细胞活性,观察盐酸阿比朵尔抗RSV的作用。结果:盐酸阿比朵尔的半数毒性浓度(TD50)为85.39 mg/L,当该药浓度为25 mg/L时,药物抗病毒生物合成组、药物直接作用组、药物抗吸附4 h组、药物抗病毒吸附8 h组的病毒抑制率分别能达到81.61%,79.57%,40.39%和78.60%。4组相应的药物半数有效浓度(ED50)为11.52,8.74,25.36和10.40 mg/L。结论:盐酸阿比朵尔在体外能明显抑制RSV引起的CPE效应,病毒抑制率随药物浓度的增加而增高,盐酸阿比朵尔能通过多种途径发挥抗RSV的作用。     

荔枝核提取物体外抗病毒活性及其机制研究

目的:研究荔枝核提取物(LSE)的体外抗病毒活性,并初步研究其抗病毒的作用机制.方法:采用细胞病变效应(CPE)减少法和MTY法检测LSE对柯萨奇B3病毒(Cox B3)、呼吸道合胞病毒(RSV)以及单纯疱疹病毒1型和2型(HSV-1和HSV-2)的体外抗病毒活性,并通过观察直接杀病毒作用、对宿主细胞的保护作用、抑制病毒吸附作用以及在病毒感染后不同时间加入实验,对LSE抑制HSV-1的作用机制进行探讨.结果:LSE有一定的细胞毒性,它不能抑制Cox B3在宿主细胞内的增殖,但对RSV、HSV-1和HSV-2表现出明显的浓度依赖性抗病毒活性,其IC<,50>值分别为32.3,27.9和20.4μg/mL.该提取物预培养Vero细胞后不能保护细胞免受HSV-1感染,但它对HSV-1有直接的灭活作用.LSE可影响HSV-1对Vero细胞的吸附,当浓度为31.3μg/mL时对病毒吸附的抑制率为51%.ISE对HSV-1在细胞内的增殖表现出很强的抑制活性.结论:LSE具有较强的体外抗RSV和HSV活性,它对HSV的抑制作用可能通过多种方式和途径实现.