鸡血藤水提物体外抗肠道病毒作用研究

目的揭示鸡血藤提取液体外抑制肠道病毒的作用。方法采用细胞病变抑制试验、MTT比色法,以病毒唑为阳性对照药,测定了鸡血藤水提液AE(aqueous extracts)对Vero E6细胞的细胞毒性,并在对细胞的最大无毒浓度下观察不同浓度的提取液AE在Vero E6细胞中对5种肠道病毒的抑制作用,计算药物半数有效浓度(50%inhibition con-centration,IC50)和药物治疗指数(therapeutic index,TI)。在此基础上采用分组实验初步探讨了提取物样品的抗病毒机制。结果提取物AE半数中毒浓度(CC50)为≥250.0 ug/ml;抑制肠道病毒柯萨奇病毒B3、B5、埃可病毒Echo 9、Echo 29、脊髓灰质炎病毒Polio virusⅠ的IC50分别为60.8、47.1、14.8、65.52、9.1 ug/ml,TI分别为4.1、5.3、16.9、3.8、8.6,对肠道病毒的抑制作用存在量效反应关系;机制研究表明,AE在与CVB3的四种分组作用方式中均呈现不同程度的抗病毒作用。结论鸡血藤抗病毒提取液AE在Vero E6细胞中对肠道病毒有明显的抑制作用;鸡血藤水提物AE不仅具有直接杀灭CVB3病毒作用,而且还可进入细胞或吸附在细胞表面,从而达到抑制或杀伤病毒的效果。     

鸡血藤水提物抗流感病毒作用的研究

目的揭示鸡血藤水提取物A.E.体外抗流感病毒A1的作用。方法采用细胞病变CPE法测定鸡血藤水提取物A.E.在MDCK细胞的抗流感病毒的活性。同时采用血凝滴度测定鸡血藤水提物A.E.对鸡胚培养甲型流感病毒的抑制作用。小鼠经滴鼻感染流感病毒造成肺炎模型,观察其肺指数、死亡率及病理切片。结果鸡血藤水提物A.E.在MDCK细胞中对流感病毒A1具有一定的抑制作用,其半数抑制浓度IC50为(74.5±4.7)μg/ml,治疗指数TI为13.3。在鸡胚中与病毒对照组比较,鸡血藤水提物A.E.各剂量组的血凝滴度降低,差异有统计学意义(P0.01)。每日灌注鸡血藤水提物A.E.能够明显降低病毒感染小鼠的死亡率、肺指数、肺病理切片坏死和浸润的程度。结论鸡血藤水提物对流感病毒A1具有抑制作用。     

上海市2002年病毒性脑炎病因病毒的分离和鉴定

采集上海市 2 0 0 2年 6 9例病毒性脑炎散发病人的脑脊液 (CSF)标本 ,用Hep 2、RD、Vero细胞分离出肠道病毒(EV) 2 1株 ,经用标准血清鉴定为柯萨奇病毒B5(Coxsackie ,Cox B5) 11株 ,Cox B2 1株 ,Cox A161株 ,埃可病毒 (E CHO) 3株 ,非脊髓灰质炎肠道病毒 (NPEV) 5株 ,Cox B5病毒分离率为 16 %。本次调查结果表明 ,Cox B5病毒是上海市 2 0 0 2年病毒性脑炎的主要病原。     

三种植物提取物对醛糖还原酶活性的抑制作用

目的分别观察黄柏、生甘草、野菊花的乙醇提取物对醛糖还原酶活性的抑制作用。方法从正常SD大鼠晶状体提取制备醛糖还原酶，体外实验以槲皮素为阳性对照，比较其与三种植物乙醇提取物对醛糖还原酶活性的抑制作用。结果随着三种植物乙醇提取物和槲皮素浓度的增加，醛糖还原酶活性逐渐降低。槲皮素IC50=68．23ug／ml，黄柏提取物IC50=107．34ug／ml，生甘草提取物IC50=185．64ug／ml，野菊花提取物IC50=118．09ug／ml，提示三种植物的乙醇提取物对AR均有抑制作用，抑制强度大小依次为槲皮素〉黄柏提取物〉野菊花提取物〉生甘草提取物。结论三种植物乙醇提取物均有抑制醛糖还原酶活性的作用，其抑制活力黄柏〉野菊花〉生甘草。     

中药苦碟子抗呼吸道病毒药效观察

目的 通过细胞内培养和小鼠体内试验观察苦碟子抗呼吸道病毒感染细胞的作用和活体内抗呼吸道病毒感染的效果。方法 体外试验采用病毒细胞病变观察（CPE）法测定药物的半数抑制浓度（IC50）和最小有效浓度（MIC）及治疗指数（TI）；体内试验采用病毒接种致肺炎动物模型试验，观察肺病变并计算病变率和病变抑制率。结果 苦碟子对呼吸道流感病毒H3N2、合胞病毒（RSV）和腺病毒（ADV-3）的治疗指数分别为10，32和16。不同剂量组对3种病毒肺病变抑制率均高于双黄莲对照组（P〈0．05）。结论 苦碟子对呼吸道流感病毒H3N2、合胞病毒（RSV）和腺病毒（ADV-3）均有良好的抑制作用。     

中医药治疗柯萨奇病毒引起的病毒性心肌炎的研究进展

正病毒性心肌炎(viral myocarditis,VMC)是心肌局限性或弥漫性的炎性病变可呈急性或慢性病程。多种病毒可诱导病毒性心肌炎发病,其中以肠道病毒中柯萨奇组病毒型(Coxsackie virusB,CVB3)最为常见,被认为是导致病毒性心肌炎的最主要原因~([1]),柯萨奇病毒B组3型(coxsackie virus B3,CVB3)属于小核糖核酸病毒科(picornaviridae),肠道病毒属(enterovirus),通常感染后会使人出现发热、打喷嚏、咳嗽等感冒症状,但也可导致新生儿、婴幼儿以及免疫缺陷的成人出现严重的心脏、胰腺及中枢神     

氯化钴对小鼠腹腔巨噬细胞的抑制作用

目的观察氯化钴（CoCl2）对巨噬细胞体外培养的抑制作用。方法采用小鼠巨噬细胞体外培养法在不同浓度（0．5、5．0、50．0mg／m1）的CoCl2作用下,应用细胞增殖抑制试验（MTT法）测定体外巨噬细胞的抑制率、应用吖啶橙染色法测定细胞凋亡情况和应用考马斯亮蓝染色法测定总蛋白含量的变化。结果50．0mg／ml的CoCl：作用24h,小鼠腹腔巨噬细胞中蛋白质相对含量为75．2％,与对照组的100％相比,差异有统计学意义（P〈0．01）;5．0和50．0mg／mlCoCl2作用48和72h,巨噬细胞中蛋白质含量与对照组相比,差异均有统计学意义（P〈0．01）。采用吖啶橙染色法测定细胞凋亡发现在CoCl2的作用下有明显凋亡现象。结论CoCl2对小鼠体外巨噬细胞增殖有明显的抑制作用,且有促细胞凋亡和抑制蛋白生成作用。     

人巨细胞病毒临床药敏检测及实验株耐药模型构建

目的测定广州部分地区造血干细胞移植术后人巨细胞病毒（HCMV）临床分离株对更昔洛韦（GCV）的敏感性,并初步建立实验室药物诱导HCMV耐药株。方法收集造血干细胞移植术后发生HCMV感染而接受GCV治疗的患者血液或尿液标本,采用HELF细胞培养法分离并鉴定HCMV临床株,50％终点法测定各临床毒株的感染性滴度TCID50,MTT法测定各毒株对更昔洛韦的敏感性（IC50）。将HCMVAD169株在含有不同浓度（1．5—300μmol／L）的GCV中作连续传代培养,分别在病毒与1．5、3、6、10、50、300μmol／L浓度GCV培养后测定其IC50结果成功分离8株HCMV临床株,经培养后测定其TCID50分别为：10^-4.12／0．1mL、10^-4.29／0．1mL、10^-4.30／0．1mL、10^-4.40／0．1mL、10^-4.42／0．1mL、10^-4.50,／0．1mL、10^-4.52／0．1mL、10^-4.62／0．1mL;8株临床HCMV分离株对GCV的半数病毒抑制浓度IC50分别为1．89—5．95μmol／L。GCV诱导HCMV实验株发现,培养液中GCV浓度增至10μmol／L时,病毒开始出现耐药,IC50为18．15μmol／L,当GCV浓度达到300μmol／L时,300μmol／L诱导培养的病毒对GCV的IC50则高达327．10μmol／L,为标准株HCMVAD169（IC50=1．88μmol／L）的174倍。结论该实验HCMV临床分离株对更昔洛韦敏感性均较高,未发现临床耐药株;通过体外药物诱导,初步构建HCMV耐药株,为HCMV的耐药研究创造条件。     

应用4种方法检测肠道病毒的结果分析

目的 对105例心肌梗塞患者和40例对照组应用4种方法检测肠道病毒（EV）。方法 用逆转录聚合酶链反应（RT－PCR）检测肠道病毒核糖核酸（EV－RNA），ELISA法检测柯萨奇B组病毒（Cox B V)IgM抗体，常规方法分离病毒及微量中和抗体（NT）。结果 病例组EV－RNA阳性23例（21．9％）。对照组1例（2．5％）；CoxB V IgM阳性29例（27．6％），对照组1例（2．5％）；病毒分离得到病毒6株（5．7％），对照组未分离出病毒；CoxB V NT阳性25例（23．8％），对照组3例（7．5％）。结论 用RT－PCR检测EV－RNA优于病毒培养，RT－PCR方法敏感，特异，简单，易于推广。采用RT－PCR，病毒培养，ELISA，NT4种方法同步进行检测EV则可以提高病毒的阳性检出率。     

Survivin在冬凌草甲素介导下对人前列腺癌细胞PC-3凋亡的作用

目的观察冬凌草甲素对人雄激素非依赖性前列腺癌细胞株-PC-3细胞的诱导凋亡作用,探讨Survivin在此过程中的作用。方法用不同浓度的冬凌草甲素干预PC-3细胞,MIT试验分析观察其对PC-3细胞活力的影响;通过用流式细胞仪分析PC．3早期凋亡细胞的百分率;用Western印迹检法、实时荧光定量PCR方法检测PC-3细胞Survivin的蛋白和mRNA表达的变化。结果（1）细胞生长抑制力呈一定的时间、剂量依赖性,冬凌草甲素浓度为2．5、5、10、20、40μmol／L时,干预48h后相对应的平均细胞生长抑制率依次为9．2％、25．3％、39．3％、77．2％、92．5％,药物抑制PC-3细胞活力的IC50约为10．29μmol／L;流式细胞仪检测经不同浓度的冬凌草甲素（0,10,20,40μmol／L）干预48h后,PC-3细胞的早期凋亡率分别为4．8％,15．4％,19．5％和27．4％（P〈0．05）。（2）冬凌草甲素以浓度依赖性方式抑制PC-3细胞的Survivin的蛋白和mRNA表达。结论冬凌草甲素能以浓度依赖性方式诱导PC-3细胞凋亡。冬凌草甲素通过影响Survivin的表达来诱导PC-3细胞凋亡。     

柯萨奇B组病毒的血清学监测

本文报道应用微量中和试验检测163例健康人群的柯萨奇病毒B组3、4、5型的中和抗体水平。中和抗体阳性率以B₄为最高(92.02%),B₃次高(62.57%), B₅最低(29.45%). B₃中和抗体几何平均滴度为14.19, B₄为102.8, B₅为3.96。B₃中和抗体滴度以1~3岁组最高,以后随年龄增长而降低。B₄中和抗体滴度除19~45岁组略高外,其余各组抗体滴度很低。     

Effect of Foeniculum Vulgare Aqueous and Alcoholic Seed Extract against Zoonotic Cutaneous Leishmaniasis

BackgroundCutaneous leishmaniasis is considered one of the major neglected tropical diseases. Drug resistance, limitary efficacy, and severe side effects remain a challenge for treatment. Foeniculum vulgare is known as a medicinal plant belonging to the Apiaceae, and anti-microbial properties of this plant have already been confirmed.MethodThe F.vulgare sterile aqueous and alcoholic extracts were prepared. In vitro has used RAW 264.7 cell line and L. major parasite (MRHO/IR/75/ER). Cytotoxicity assay on macrophages (CC50), cytotoxicity assay on promastigotes (IC50), and cytotoxicity assay on infected macrophages (EC50) were accomplished with both extracts by MTT and light microscopy methods. Four in vivo were allocated in four groups and five BALB/c mice each group. Stationary phase promastigotes were inoculated into the base of mice tails subcutaneously (SC). Measurement of the body weight, lesion size, parasite burden of the lesion, and spleen after 4 weeks for evaluation effects of the alcoholic extract on CL was done.ResultsThe results of in vitro revealed that the optimal concentrations of both extracts reducing the promastigotes and amastigotes growth. Alcoholic extract no harmful side effects for the host macrophages, while were indicated has a potent action against L. major. In vivo results after 4 weeks did not show any variation in lesion size and body weight. Also, lesion size and spleen parasite burden decreased in comparison to no treatment group.ConclusionThe alcoholic extract could be a new alternative treatment for cutaneous leishmaniasis. However this extract needs more investigation for novel herbal drugs against CL.     

用L-hCAR细胞选择性地分离柯萨奇B组病毒

为研究L -hCAR细胞对柯萨奇B组病毒 (CoxsackievirustypeB ,Cox B)的敏感性和特异性 ,以及用L -hCAR细胞从粪便标本中分离Cox B的情况 ,将 43个肠道病毒标准株同时接种于Hep -2和L -hCAR细胞 ,在L -hCAR细胞上只有Cox B增殖。分别用Hep -2及L -hCAR细胞对Cox B 1～ 6型标准株进行滴定 ,两种细胞上的病毒滴度无显著性差异。在病毒分离试验中 ,用 94个已知为非脊髓灰质炎肠道病毒的粪便标本分别接种于Hep -2和L -hCAR细胞 ,在 14天的观察期间内 ,共有8份粪便标本在L -hCAR细胞上产生细胞病变效应。经鉴定 ,8份均为Cox B。结果表明L -hCAR细胞对Cox B敏感且特异 ,能用L -hCAR细胞选择性地从粪便标本中分离Cox B     

Coffee extracts inhibit platelet aggregation in vivo and in vitro

The effect of aqueous coffee extracts on platelet aggregation in humans (in vitro) and rabbits (both in vitro and in vivo) was investigated. Coffee extracts were found to have anti-aggregatory effects on in vitro platelet aggregation induced by ADP or arachidonate but not by collagen. Coffee extracts were also effective after intravenous administration in rabbits. The compound(s) responsible for these effects are water-soluble, heat-resistant, appeared to be different from salicylates, and might also be due to unidentified compounds besides nicotinic acid or known xanthines. Coffee extract and selected fractions decreased the conversion of [14C]-arachidonic acid to thromboxane B2 by the platelets. These studies show that coffee extracts contain compounds which are active in inhibiting platelet aggregation, a critical step involved in thrombosis and other vascular disorders.     

浙江省1996～1999年急性弛缓性麻痹病例中非脊髓灰质炎肠道病毒分析

浙江省1996～1999年急性弛缓性麻痹（AFP）病例粪便标本中非脊髓灰质炎（脊灰）肠道病毒（NPEV）的平均分离率为11.2％。在67株NPEV中,柯萨奇病毒（Coxsackievirus,Cox.）B组21株,占31.3％;埃可病毒（EntricCytopothicHumanOrphanviruses,ECHO）25株,占37.4％;不能确定型别21株,占31.3％。NPEV按月分布显示,Cox.B病毒高峰期在5～7月,在1～3、10、12月未分离到;ECHO病毒全年均有,高峰期在8～11月。Cox.B病毒在Hep－2、RD细胞上平均滴度分别为10     

Neuroprotective effects of several korean medicinal plants traditionally used for stroke remedy

The neuroprotective effect of six aqueous extracts and one alcoholic extract prepared from seven medicinal plants that have been recorded as having therapeutic effects for stroke in Korean traditional medicine were studied using both in vitro and in vivo cerebral ischemia models. Among the extracts tested, the aqueous extracts of Acorus gramineus, Chrysanthemum indicum, and Pinus densiflora and the alcoholic extract of Vitis vinifera significantly increased the cell viability of SK-N-SH human neuroblastoma cells exposed to oxygen-glucose deprivation (P < .05). Following two-vessel occlusion in gerbils, extracts of P. densiflora and V. vinifera significantly increased the number of surviving cells/mm(2) of the CA1 region by 2.1-2.2-fold (P < .01). Oral or intraperitoneal administration of S-allyl cysteine, as a positive control, also markedly increased cell survival up to about 3.3-fold at the dosage of 300 mg/kg of body weight (P < .01). These results indicate that P. densiflora and V. vinifera exert neuroprotective effects against ischemic insults in both the in vitro and in vivo cerebral ischemia models and prompted us to further characterize the detailed mechanism of action and elucidate the active principles.     

Attempts to infect pigs with Coxsackie virus type B5

Despite the existence of a close serological relationship between the enteroviruses Swine Vesicular Disease (SVD) and Coxsackie type B5 (Cx B5), the administration of this Coxsackie virus type to susceptible pigs by various routes failed to produce clinical disease.     

Synthesis of some novel thiourea derivatives obtained from 5-[(4-aminophenoxy)methyl]-4-alkyl/aryl-2,4-dihydro-3H-1,2,4-triazole-3-thiones and evaluation as antiviral/anti-HIV and anti-tuberculosis agents

As a continuation of our previous efforts on N-alkyl/aryl-N'-[4-(4-alkyl/aryl-2,4-dihydro-3H-1,2,4-triazole-3-thione-5-yl)phenyl]thioureas 1-19 and N-alkyl/aryl-N'-[4-(3-aralkylthio-4-alkyl/aryl-4H-1,2,4-triazole-5-yl)phenyl]thioureas 20-22, a series of novel 5-[(4-aminophenoxy)methyl]-4-alkyl/aryl-2,4-dihydro-3H-1,2,4-triazole-3-thiones 23-26 and several related thioureas, N-alkyl/aryl-N'-{4-[(4-alkyl/aryl-5-thioxo-4,5-dihydro-1H-1,2,4-triazol-3-yl)methoxy]phenyl}thioureas 27-42 were synthesized for evaluation of their antiviral potency. Structures of the synthesized compounds were confirmed by the use of (1)H NMR, (13)C NMR and HR-MS data. All compounds 1-42 were evaluated in vitro against HIV-1 (IIIB) and HIV-2 (ROD) strains in MT-4 cells, as well as other selected viruses such as HSV-1, HSV-2, Coxsackie virus B4, Sindbis virus and Varicella-zoster virus using HeLa, Vero, HEL and E6SM cell cultures, and anti-tuberculosis activity against Mycobacterium tuberculosis H37Rv. Compounds 4 and 5 showed weak activity against HSV-1, HSV-2 and TK(-) HSV, whereas eight compounds showed marginal activity against Coxsackie virus B4. The most active derivative in this series was compound 38 which showed moderate protection against Coxsackie virus B4 with an MIC value of 16 microg/ml and a selectivity index of 5. This compound was also active against thymidine kinase positive Varicella-zoster virus (TK(+) VZV, OKA strain) with an EC(50) value of 9.9 microg/ml. Compound 38 was the most active compound with 79% inhibition against M. tuberculosis H37Rv.     

亚硒酸钠对柯萨奇B_5病毒在细胞内复制的影响

作者研究了Na_2SeO_3对柯萨奇B_5病毒在细胞内复制的影响。通过向营养液中加入一定剂量的Na_2SeO_3培养细胞,使细胞传至3～10代,柯萨奇B_5病毒在细胞内的增殖量降低,而细胞存活率明显增高,与对照组相比,差异显著。这种抑制病毒增殖作用的原因,不是Na_2SeO_3对病毒的直接灭活作用,而可能是Na_2SeO_3使细胞内发生了某些代谢改变,达到了一定程度地抑制病毒增殖的作用。     

The role of coxsackieviruses infection in the children of insulin dependent diabetes mellitus

In the present study, there were two groups of diabetic patients. The first group was newly diagnosed diabetic patients of less than one year duration of disease. They were 40 patients. The second group was diabetic patients with more than one year duration of disease. They were 30 patients. The control group was 30 normal healthy children. Evidence of virus infection was detected by tissue culture isolation, neutralization test, RT-PCR, IgM, and IgG specific antibodies for coxsackie B viruses. There was significant increase in percent of tissue culture isolation of EV in group I more than group II while the percent in the two groups were significantly increased than the control group. Identification of the type of EV by neutralization test revealed that most of the type cases were Coxsackie B4 virus and one case was Coxsackie B6 virus (the most important diabetogenic strain). Viral RNA detection by PCR was done and revealed that most of cases in group I diabetic patients were positive for enterovirus while one case in group II of diabetic patients was positive. As regards Coxsackie virus B IgM antibodies positivity, there was increase in the percent in group I than group II, as it is a marker of acute infection. As regards Coxsackie virus B IgG antibodies positivity, there was no significant difference between group I and group II of diabetic patients, as it is a marker of past infection and it persists for years after the first episode.