甘露消毒丹及其拆方体外抗肠道病毒71型作用

目的观察甘露消毒丹及其拆方体外抗肠道病毒71型(EV71)的作用。方法运用细胞培养技术,病毒毒力测定采用Reed-Muench法计算半数组织培养感染量(TCID50),确定实验用为100 TCID50。将甘全方(甘露消毒丹全方)、清残方(甘露消毒丹方中清热解毒药物组成)、利残方(甘露消毒丹方中化浊利湿药物组成)、病毒唑药液用维持液1∶8、1∶16、1∶32、1∶64、1∶96、1∶128、1∶256稀释,采用MTT法测定各浓度药物吸光度OD值,根据各药不同浓度的OD值计算其细胞存活率,以药物浓度为横坐标(g/ml)、细胞存活率(%)为纵坐标绘出非线性回归曲线,选择各药的90%细胞存活率浓度为药效学实验浓度。分别将90%细胞存活率浓度的甘全方、清残方、利残方、病毒唑药液和100 TCID50病毒各50μl混合,加到单层人恶性胚胎横纹肌瘤细胞(RD细胞)上,各设4复孔,每孔均设正常细胞对照、病毒对照,采用MTT法计算各药物对病毒生长的抑制率。结果根据Reed-Muench法计算得出TCID50=10-2.89/ml,从回归曲线获得各药90%细胞存活率浓度分别为甘全方:10-2.43=0.0037 g/ml;清残方:10-2.7=0.0020 g/ml;利残方:10-2.09=0.0081 g/ml;病毒唑:10-3.19=0.64 mg/ml。清残方EV71病毒抑制率为97.4%,甘全方为91.74%,利残方为75.64%,病毒唑为36.99%。结论甘露消毒丹全方及其拆方具有比病毒唑更强的抗EV71作用,清热解毒法在方中发挥了更强的抗EV71作用。     

甘露消毒丹对CAV16、EV71感染小鼠肠道微生物的影响

目的:研究甘露消毒丹对CAV16、EV71感染小鼠肠道微生物的影响。方法:将33只小鼠随机分为3组,每组各11只。正常组灌胃蒸馏水;实验1组先灌胃甘露消毒丹,再腹腔注射CAV16;实验2组先灌胃甘露消毒丹,再腹腔灌射EV71。取小鼠肠道微生物进行分析。结果:与正常组比较,实验1组、实验2组细菌总数、双歧杆菌总数、乳酸杆菌总数、大肠杆菌总数均有所提高,其中双歧杆菌、乳酸杆菌、大肠杆菌总数差异有统计学意义(P0.01,P0.05)。结论:甘露消毒丹能促进肠道益生菌的生长,改善肠道微生态平衡,预防病毒CAV16、EV71对机体的损害。     

紫萁贯众提取物体外抗病毒实验研究

目的:探讨紫萁贯众提取物对肠道病毒71型(EV71)及人乙肝病毒(HBV)的体外抗病毒效果。方法:应用体外细胞培养及抗病毒实验观察紫萁贯众对肠道病毒EV71及HBV抑制作用。结果:紫萁贯众水提物具有显著的抗EV71的活性,其抑毒指数为64,并且水提煮过的效果优于冷处理者;丙酮提取物实验细胞全部脱落,说明丙酮提取物能显著抑制HBV复制。结论:紫萁贯众水提物具有显著的抗肠道病毒71型效果;丙酮提取物具有良好的抗乙型肝炎病毒的作用。     

甘露消毒丹及其拆方对感染肠道病毒71型细胞miR-146a和Toll样受体4mRNA表达的影响

目的探讨甘露消毒丹及其拆方对肠道病毒71型(EV71)感染后免疫及炎症反应的调节作用。方法运用细胞培养技术,设甘露消毒丹组(甘全方组)、清残方组、利残方组、病毒唑组、正常组、模型组,进行病毒毒力和药物细胞毒性测定后,以药物作用于EV71感染后的RD细胞,24 h后RT-PCR检测各组细胞miR-146a、Toll样受体4(TLR4)mRNA表达。结果与正常组比较,模型组miR-146a mRNA表达降低,TLR4 mRNA表达升高;与模型组比较,甘全方组、清残方组、利残方组miR-146a mRNA表达升高、TLR4 mRNA表达降低,病毒唑组与模型组比较差异不明显;与甘全方组比较,清残方组miR-146a、TLR4 mRNA表达降低,利残方组miR-146a mRNA表达升高、TLR4 mRNA表达降低;与清残方组比较,利残方组miR-146a mRNA表达升高,TLR4 mRNA表达无明显差异。结论甘露消毒丹可能通过调节miR-146a、TLR4 mRNA表达从而影响EV71感染所致免疫反应,其中清残方与利残方可能存在互相拮抗效应。     

甘露消毒丹对柯萨奇病毒体外抑制作用的实验研究

目的：观察甘露消毒丹全方、残方及加味方对柯萨奇病毒的体外抑制作用。方法：采用组织培养技术，在培养细胞中观察甘露消毒丹全方、残方及加味方水煎液分别对培养细胞的最高无毒浓度、对柯萨奇病毒增殖产量的影响以及在不同时间内抑毒效果。结果：甘露消毒丹全方、残方及加味方对细胞的最高无毒浓度均为１：４０（即２５ｇ／Ｌ），该浓度对柯萨奇病毒Ｂ２、Ｂ３、Ｂ４株在培养细胞内的增殖有明显的抑制作用，抑制指数均〉２；全方对     

疏风解毒颗粒防治手足口病的体内外药效学研究

目的:通过观察疏风解毒颗粒在体内对手足口病病毒EV71感染BALB/c乳鼠模型的保护作用和在体外对EV71等肠道病毒的抑制作用,评价疏风解毒颗粒防治手足口病的药效.方法:采用疏风解毒颗粒3岁儿童临床2倍、等倍和1/2倍剂量(14.08,7.04,3.52 g·kg-1)分别对手足口病病毒EV71H株感染的BALB/c乳鼠手足口病动物模型进行预防性给药(ig,3d)和治疗性给药(ig,5d),观察乳鼠感染的严重程度、死亡数,计算动物死亡率和生命延长率;采用疏风解毒颗粒在体外对Vero细胞无明显毒性的5个剂量,对人手足口病病毒EV71H株,柯萨奇病毒B3株,B4株,B5株的致Vero细胞病变的抑制作用进行观察.结果:疏风解毒颗粒对手足口病病毒EV71感染的乳鼠模型具有明显的预防和治疗作用,可降低感染程度和死亡率,具有明显的保护和延长生命的作用;疏风解毒颗粒在无明显毒性的浓度下,在体外对人手足口病病毒EV71 H株、CoxB3株、CoxB5株均具有明显的抑制作用.结论:疏风解毒颗粒在体内外均具有较好的防治手足口病的作用.     

大蒜素体外抗肠道病毒EV71的研究

目的:探究肠道病毒EV71对VERO细胞的损害作用和大蒜素对肠道病毒EV71的治疗性和预防性干预作用。方法:分别观察大蒜素对VERO细胞的毒性,肠道病毒对VERO细胞的毒性,大蒜素对肠道病毒EV71的治疗性和预防性干预作用。结果:大蒜素对VERO细胞的半数中毒浓度(TC50)为724.44μg/ml,最大无毒浓度(TC0)为150μg/ml;大蒜素对肠道病毒EV71的治疗性干预试验的半数有效浓度(EC50)为16.22μg/ml,治疗指数(TI)为44.66;大蒜素对肠道病毒EV71的预防性干预试验的最佳作用时间为6h,此时大蒜素的半数有效浓度(EC50)为39.64μg/ml,治疗指数(TI)为18.28。结论:大蒜素体外对肠道病毒EV71有治疗性和预防性干预作用。     

甘露消毒丹的来源与现代应用概况

<正>近年来,甘露消毒丹在临床和实验中应用很多,甘露消毒丹在抗病毒方面的实验研究颇多,主要集中在抗感染性疾病方面。本文就甘露消毒丹的来源与现代临床和实验研究作一综述。1来源甘露消毒丹究竟来源何处,近年来,医家对甘露消毒丹的出处众说纷纭。现在主要有两种说法:一是甘露消毒丹出于《续名医类案》,二是甘露消毒丹出于《医效秘传》。查阅《续名医类案·卷五·疫证》~([1]):"雍正癸丑,疫气流行,抚     

甘露消毒丹治疗急性病毒性肝炎临床观察

甘露消毒丹、三仁汤两方,均有清热利湿的作用,临床常用于病毒性肝炎的治疗,为了观察两方治疗病毒性肝炎的疗效,通过对７６例急性病毒性肝炎的临床观察,结果显示,甘露消毒丹在改善症状、体征、恢复肝功能及ＨＢｓＡｇ、ＨＢｅＡｇ转阴方面均优于三仁汤。     

中药抗肠道病毒71型感染研究现状北大核心CSCD

肠道病毒71型(EV71)是引起手足口病的主要病原体,因其能引起中枢神经系统并发症,甚至肺出血、肺水肿,导致婴幼儿死亡而引起国内外广泛关注。然而,至今仍没有上市的疫苗和有效的抗病毒疗法。传统中药在抗病毒治疗上以其天然、安全、毒副作用少而被众多专家青睐。临床研究已验证中药在EV71感染中的治疗作用,比如清开灵、喜炎平以及蓝岑口服液等。在临床治疗的基础上,众多学者致力于研究中药抗EV71病毒的有效性及其有效成分,如荆芥、鱼腥草等中草药被发现在体外有明显的抗病毒活性。本文就EV71引起的手足口病的中药临床治疗及中药抗EV71病毒活性的基础研究现状进行综述。     

In vitro activity of Paris polyphylla smith against enterovirus 71 and coxsackievirus B3 and its immune modulation

Enterovirus 71 (EV71) and coxsackievirus B3 (CVB3) have resulted in severe pathogenesis caused by the host's immune response, including the cytokine cascade. Paris polyphylla Smith is a folk medicinal plant in Asia traditionally prescribed for the reduction of pain and elimination of poisoning. In this study, we investigated the anti-EV71 and CVB3 activity of P. polyphylla Smith as well as its immune modulation. The IC(50) for the P. polyphylla Smith 95% ethanol extract against EV71 and CVB3 were 12.5-23% and 99-156% of that of ribavirin, a positive control. Prevention of viral infection, viral inactivation, and anti-viral replication effects against both EV71 and CVB3 were demonstrated by the extract, the anti-viral replication effect being dominant. The extract significantly increased IL-6 production in both EV71- and CVB3-infected cells. A high correlation was possibly demonstrated between the high amounts of IL-6 induction in the EV71 and CVB3-infected cells and the anti-viral replication activity of the extract. In conclusion, good anti-EV71 and CVB3 activity was observed in the P. polyphylla Smith 95% ethanol extract. The high amounts of IL-6 induction in the virus-infected cells played a key role in the anti-viral activity of the extract.     

羊栖菜提取物的体外抗病毒作用

目的:探讨羊栖菜提取物的体外抗病毒作用。方法:对羊栖菜的提取物进行了常见的第三类病原体病毒呼吸道合胞病毒(RSV)、单纯疱疹病毒1型(HSV-1)、肠道病毒71型(EV71)、柯萨奇病毒A_(16)、柯萨奇病毒B_2、柯萨奇病毒B_3、柯萨奇病毒B_5(CVA_(16)、CVB_2、CVB_3、CVB_5)等7种病毒的抗病毒效应研究,显微镜下观察细胞存活率。结果:羊栖菜提取物对上述7种病毒的抑制能力远远优于广谱抗病毒药物利巴韦林,尤以对EV71的抑制效果最为明显,抑毒指数极高。结论:羊栖菜提取物对这7种病毒均具有良好的抑制作用,为进一步研究羊栖菜提取物对第三类病原体病毒的广谱抗病毒作用奠定基础。     

中药抗EV71病毒活性及其在治疗手足口病中的应用

手足口病是由肠道病毒引起的传染病.柯萨奇病毒A16和肠道病毒71型(EV71)是本病的主要致病原,且后者较凶险,更易导致严重并发症而致死、致残,危害性较大.目前对EV71感染尚缺乏有效的药物,因此研制抗EV71病毒活性的药物意义重大.中国中药资源丰富,具有进一步开发和利用的独特优势,从中筛选出具有抗EV71病毒活性的成分,进而开发出新的抗病毒药物具有很大潜力.同时,我国自1981年在上海始见手足口病以来,中药在临床治疗手足口病方面的报道日益增多且方法多样,大多疗效满意,对提高临床诊治水平、控制疾病的发生发展有着积极作用,展现了其独特的临床优势与特点.     

清热化湿法治疗慢性支气管炎急性发作期临床观察

目的观察清热化湿法对慢性支气管炎急性发作期(以下简称慢支急发)临床疗效的影响。方法57例慢支急发(痰湿壅肺兼湿热蕴脾型)患者随机分为2组。对照组27例予西医常规治疗,治疗组30例在对照组基础上加服甘露消毒丹汤剂。治疗14日后,比较2组临床疗效和主要症状、体征、血气的变化。结果治疗组控显率高于对照组;治疗组咳嗽、咯痰、纳呆、腹胀、恶心等症状的改善及动脉血氧分压(PaO2)、动脉二氧化碳分压(PaCO2)的改善均优于对照组,有统计学意义(P<0.05)。结论清热化湿法与西药联合治疗慢支急发有较好的临床疗效。     

Glycyrrhizic acid as the antiviral component of Glycyrrhiza uralensis Fisch. against coxsackievirus A16 and enterovirus 71 of hand foot and mouth disease

Ethnopharmacological relevance

The radices of Glycyrrhiza uralensis Fisch. and herbal preparations containing Glycyrrhiza spp. have been used for thousands of years as an herbal medicine for the treatment of viral induced cough, viral hepatitis, and viral skin diseases like ulcers in China. Glycyrrhizic acid (GA) is considered the principal component in Glycyrrhiza spp. with a wide spectrum of antiviral activity.

Aim

The present study attempt to validate the medicinal use of Glycyrrhiza uralensis for hand, foot and mouth disease (HFMD) and further to verify whether GA is an active antiviral component in the water extract of Glycyrrhiza uralensis.

Materials and methods

Radices of Glycyrrhiza uralensis Fisch. were extracted with hot water. The chemical contents of the extract were profiled with HPLC analysis. The antiviral activity of the extract and the major components was evaluated against infection of enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) on Vero cells. The cytopathic effect caused by the infection was measured with MTT assay. Infectious virion production was determined using secondary infection assays and viral protein expression by immunoblotting analysis.

Results

The extract at 1000 μg/ml suppressed EV71 replication by 1.0 log and CVA16 by 1.5 logs. The antiviral activity was associated with the content of GA in the extract since selective depletion of GA from the extract by acid precipitation resulted in loss of antiviral activity. In contrast, the acid precipitant retained antiviral activity. The precipitant at a concentration of 200 μg/ml inhibited EV71 and CVA16 replication by 1.7 and 2.2 logs, respectively. Furthermore, GA dose-dependently blocked viral replication of EV71 and CVA16. At 3 mM, GA reduced infectious CVA16 and EV71 production by 3.5 and 2.2 logs, respectively. At 5 mM, CVA16 production was reduced by 6.0 logs and EV71 by 4.0 logs. Both EV71 and CVA16 are members of Enterovirus genus, time-of-drug addition studies however showed that GA directly inactivated CVA16, while GA anti-EV71 effect was associated with an event(s) post virus cell entry.

Conclusions

This study validated the medicinal usefulness of radices Glycyrrhiza uralensis against the etiological agents of HFMD. In addition to the identification of GA as the antiviral component of Glycyrrhiza uralensis against EV71 and CVA16 infection, this study also reveals that GA inhibits EV71 and CVA16 with distinct mechanisms.     

中医药治疗手足口病48例临床观察

目的：观察中医药治疗手足口病普通型的临床疗效.方法：采用中医分证论治手足口病,治疗组给予银翘散加减或甘露消毒丹加减治疗,对照组给予利巴韦林治疗及对症治疗.两组病例均治疗5天后观察临床疗效.结果：从两组治愈率和好转率比较,治疗组优于对照组（t=2.8070,P〈0.005）.结论：应用银翘散、甘露消毒丹加减治疗手足口病,未见明显不良反应,结果显示治疗组优于对照组.     

抗病毒口服液联合利巴韦林气雾剂治疗手足口病的临床研究

目的：观察抗病毒口服液联合利巴韦林雾剂治疗手足口病的临床疗效。方法收集2012年5月至2014年1月河北省临西县人民医院儿内科手足口病患儿135例,按随机数字表法将患儿随机分为两组,对照组67例采用利巴韦林气雾剂治疗,治疗组68例在对照组治疗基础上配合口服抗病毒口服液治疗,治疗7d 后观察两组患儿临床症状的改善情况,并进行病毒检测。结果治疗组体温恢复时间[（2.51±0.42）d比（3.92±0.61）d;t＝15.621,P＜0.01]以及口腔溃疡[（3.01±0.32）d比（4.52±1.21）d;t＝9.880, P＜0.01]、皮疹消退[（4.22±1.34）d 比（6.33±1.41）d;t＝8.914,P＜0.01]时间均显著性短于对照组。治疗组痊愈率显著性高于对照组（88.2%比44.8%;χ^226.761,P＜0.01）,但总有效率与对照组无显著性差异（94.1%比83.6%;χ^22.801,P＝0.094）。治疗组通用型肠道病毒（90.9%比52.6%;χ^213.331,P＜0.01）和肠道病毒71型（83.3%比52.4%;χ^24.307,P＝0.038）转阴率显著性高于对照组,但柯萨奇病毒A16型转阴率与对照组无显著性差异（93.3%比81.8%;χ^20.082,P＝0.774）。结论利巴韦林气雾剂联合抗病毒口服液治疗可提高手足口病患儿的肠道病毒转阴率,缩短口腔溃疡和皮疹消退时间。     

阿昔洛韦联合蒲地兰消炎口服液治疗肠道病毒71型手足口病疗效观察

目的：观察阿昔洛韦联合蒲地兰消炎口服液治疗肠道病毒71型手足口病的临床疗效。方法：将100例患儿随机分为对照组50例和治疗组50例,治疗组给予阿昔洛维10mg／kg加入5％葡萄糖注射液250mi静脉滴注,每日1次;蒲地兰口服液口服,3次／d．连服6—12d,具体用量：小于等于1岁患儿每次1／3支,1-2岁患儿每次1／2支,2～4岁患儿每次2／3支,4～5岁患儿每次1支。对照纽仅给予阿昔洛维注射液治疗。两组均采用补液、对症等治疗,少数病例疑合并细菌感染者加用抗菌药物抗感染治疗,并观察两组疗效。结果：退热、咽痛消失、疱疹消退、EV71转阴及治愈时间治疗组均优于对照组（P〈0．05）。结论：阿昔洛韦联合蒲地兰口服液治疗肠道病毒71型手足口病具有较好临床疗效。     

甘露消毒丹加减治疗高胆红素血症的临床研究

目的：观察甘露消毒丹加减治疗肝炎后高胆红素血症的临床疗效。方法：将80例患者随机分为治疗组和对照组,各40例。治疗组予甘露消毒丹加减治疗,对照组予茴三硫片治疗。结果：治疗组有效率为95．0％,对照组为77．5％,两组疗效比较差异有统计学意义（P〈0．05）,治疗组优于对照组。结论：甘露消毒丹加减治疗肝炎后高胆红素血症疗效确切,副作用小,复发率低。     

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??OBJECTIVE To analyze the endogenous metabolite changes in the sera of kidney-yang deficiency syndrome mice infected with influenza virus A after intervention by ribavirin. And to explore the mechanism of pharmacological or toxicity effect of ribavirin. METHODS KM mice were randomly divided into three groups as normal group, model group and ribavirin group. Mice were infected with virus A after fifteen days Kidney-Yang deficiency syndrome was established. Ribavirin group were orally administrated with ribavirin for 6 consecutive days after inoculation, and the other two groups were given with equal volume of saline solution in the same way. Body weight, rectal temperature were recorded daily. Serum samples were collected from mouse 24 h after the last administration for HPLC-TOF/MS analysis. RESULTS The results show that ribavirin has good therapeutic effects on the lung index and high mortality rate of mice model. Compared with normal and model groups, the body weight and rectal temperature of them performed falling continuously. The LC-MS data were analyzed with multivariate statistical analysis and 14 potential metabolic markers were obtained which contained D-glucose, sphinganine, linoleic acid and so on. In ribavirin group, metabolism of linoleic acid, arachidonic acid and sphinganine appeared the trend of normal. And sugar and glycerophospholipid became disorders. CONCLUSION The metabolomics study and pharmacological experiment show that ribavirin might play a role of efficacy in a way that has close correlation with the linoleic acid, arachidonic acid and sphingolipid metabolic pathways. And the toxicity effect may be related to sugar and glycerophospholipid metabolic pathways.