Antiviral Activity of the Effective Monomers from Folium Isatidis Against Influenza Virus in Vivo

In order to evaluate the anti-influenza virus activity of the effective monomer from Folium Isatidis (FI) in vivo,we established mice model with viral pneumonia and divided them into 3 different dose groups,then observed their lung indexes,pulmonary pathological changes,pulmonary virus hemagglitination titers,living time and death rates.The results showed that the monomer could reduce the pulmonary index from 2.64 to 1.93,1.63 and 1.40 (P<0.01) and decrease the hemagglitination titer from 1.15 to 0.84,0.70 and 0.59 (P<0.01).In addition,different groups of FI could significantly lessen the mortality rate from 100% to 30%,25% and 15%,and prolong the living time from 5.1d to 6.5d,8.4d and 8.9d respectively(P<0.01).The high dose (75 mg/kg/d) has the similar effect with 100 mg/kg/d dose of virazole(P>0.05),and more effective than 200 mg/kg/d dose of antiviral liquor (P<0.05).

Antiviral Activity of GuiQi Polysaccharides against Enterovirus 71 in vitro

In this study,we have investigated the antiviral activity of GuiQi polysaccharides (GQP) upon enterovirus 71 (EV71) in vitro.An assay using methyl thiazolyl tetrazolium (MTT),and analyses of cytopathic effects (CPE)were used to examine the antiviral activity of GQP upon Vero cells infected with EV71.The results revealed that GQP at concentrations below 31.2μg/mL exhibited significant antiviral effects upon EV71 when applied under three different experimental protocols.GQP was most strongly active in preventing the adsorption of EV71 to target cells and in this respect it was significantly more effective than ribavirin.In addition,it was clear that GQP could inhibit viral replication when added to cells 2 h after infection,but if added at the point of infection its effect was weak.GQP is considered to be less toxic than ribavirin,and may warrant further evaluation as a possible agent in the treatment of hand,foot and mouth disease (HFMD).

The Effect of a Dietary Supplement Spirulina and Bifidobacterium adolescentis on the Cholesterol-Lowering in Vitro and in Vivo

The survey＇s results showed a significant percentage of women and especially men having an above normal cholesterol. In order to help reduce excessive rate of blood cholesterol, we used a local strain of Spirulina （Tamanrasset, in south of Algeria） associated with probiotic bacteria （Bf adolescentis）. Experiments in vitro showed a significant degradation of total cholesterol by the combination of Spirulina and Bf adolescentis （74.5%） after 72 hours incubation at 37 ~C. A cholesterol is added to the standard diet mice in order to increase the total cholesterol for three lots. However, the rates of total cholesterol in mice receiving lower fermented milk with Bf adolescentis enriched by dry Spirulina. This shows that the decrease of cholesterol rate is closely related to the presence of Bf adolescentis and Spirulina. In vitro and in vivo results show, however, that it is possible to obtain a natural product （Spirulina） and a pseudo-strain lactic （Bf adolescentis） and to participate in the prevention of cardiovascular disease risk factor whose hand is cholesterol.

Study on the in vitro activity of Hehuan Yin aqueous extract against hepatitis C 2a virus

Objective:To study the anti-HCV activity and mechanism of Hehuan Yin aqueous extract.Methods:Huh7.5.1 cells were used to establish the HCV2a virus infection model.Cell survival rate(%)and Renilla Luciferase Assay Kit(%)were calculated by Celltiter-GLO Assay for evaluating CC50,EC50 and SI values.To observe the drug resistance of the virus to different concentrations of Hehuan Yin within 72 hours by detecting luciferase activity,western-blot was used to detect the protein expression levels of NS5A,NS3 and NS5B.Results:the CC50,EC50 and SI of Hehuan Yin against HCV2a were 132.50g/ml,1.90g/ml and 67.90 respectively.The EC50 after 24h,48h and 72h administration were 18g/ml,5.8g/ml and 2.3g/ml respectively.Within the range of drug concentration,the aqueous extract Hehuan Yin had inhibitory effect on the expression of NS5A and NS5B proteins in a dose-effect relationship,but had no obvious effect on the expression of NS3 protein.Conclusion:The aqueous extract of Hehuan Yin may inhibit the replication of HCV2a virus by changing the protein expression levels of NS5A and NS5B,and the virus has no tolerance to the aqueous extract of Hehuan Yin.

Fermented Herbal Decoction Selectively Targeting Human Cancer Cell Line and Human Pathogenic Microorganism

Introduction: Prolonged immuno-suppressed status promised to induce internal growth of malignant cell and infectious agent, yet, only a small part of affected individuals seek medical attention or berried by commercially over-flowed fake information. Several studies have described complementary and alternative medicine as effective strategies for improving anti-infectious agent including malignant cell. The purpose of this study was to investigate the effect of a fermented herbal decoction (FHD) both in vitro and in vivo to malignant cells and microorganism by regulating leukocyte subset proportioning FHD as dietary material. Methods: In this approach of alternative study, selective anti-cancer effect by fermented decoction was tried to show first in vitro system both, cancer cell and virus strain. The fermented herbal decoction consisting of 80 sorts of herbs and fruits. The selective toxicity was set up and then for immunological factors in animal and human. The most important factor is to reduce side effect for a normal cell. Results: First, FHD was proved as safe by animal test. FHD regulated also the proportion of granulocyte and lymphocyte ratio both animal and human. In vitro culture showed selective toxicity by FHD against human melanoma and leukemia cell line but reduced toxicity was showed by normal cell line. As for the anti-virus activity, anti-virus effect was tested on the feeder layer of human fibroblast cell, after 9 days of culture. Second, FHD inhibits colon cancer growth in 3-methylholanthrene induced cancer in rat. Conclusion: The present results suggest that our fermented herbal decoction showed selective anti-cancer activities and anti-virus activities, together with the regulative effect on the immune system.

Therapeutic Activity of Partially Purified Fractions of Emblica officinalis （Syn, Phyllanthus embfica） Dried Fruits against Trypanosoma evansi

Emblica officinalis （E. oJficinalis） dried fruits were evaluated for its antitrypanosomal activity and cytotoxic effects. Vero cell line maintained in DMEM （Dubecco＇s Modified Eagle Medium） and incubated with Trypanosoma evansi for more than 12 h. MPE was added to the Vero cell culture medium at different concentrations （250-1,000 μg/mL） with trypanosomes concentration （1 × 106 trypanosomes/mL in each ELISA plate well） and incubated at appropriate conditions for 72 h. In-vitro cytotoxieity of MPE of E. officinalis was determined on Vero cells at concentrations （（1.56-100 ~tg/mL）. Acute toxicity and in-vivo infectivity tests were done in mice. Obtained MPE ofE. officinalis underwent process of purification via column chromatography, preparative chromatography and HPLC （higher performance liquid chromatography） with bioassay at different strata on Alsever＇s medium. In-vivo assay for trypanocidal activity, MPE and PPFs （partially purified fractions） of E. officinalis with two sets of mice, each mouse was inoculated with 1 × 104/mL oftrypanosomes and treated （48 h post inoculation） at concentrations （12.5, 25, 50, 100 and 200 mg/kg body weight） were administered at dose rate of 100 [tL per mouse via intraperitoneal route （in treating parassitemic mice） to different groups of mice, 6 mice per concentration. HPLC of partially purified fractions ofE. officinalis was carried out with mobile phase ofacetonitdle： water （40：60） in gradient mode. In vitro, MPE induced immobilization and killing of the parasites in concentration-time dependent manner. Significant reduction of trypanosomes counts from concentration of 250μg/mL and complete killing of trypanosomes at 5th hour of observation, which was statistically equivalent to 4th hour of Diminazine Aceturate （Berenil）, standard reference drug used. HPLC of the partially purified fractions revealed two major prominent peaks at retention time of 1-4 min. In vivo, both MPE and PPFs of test material did prolong lives of mice by 6-9 days but could not cure them. At concentration of 2,000 kg/kg body weight of MPE in acute test, all mice survived. For in-vivo infectivity test, mice injected with immobilized trypanosomes developed parasitemia and died while, the other group survived. MPE, PPFs and Diminazine Aceturate were toxic to Vero cells at all concentrations exception of 1.56, 1.56-3.13 and 1.56-6.25 μg/mL, respectively. From this report, PPFs ofE. officinalis dried fruits demonstrated potential pathway for a new development oftrypanocide in near future if additional investigations are put in place.

In Vivo and in Vitro Antiviral Effects of Berberine on Influenza Virus

Objective：To explore the potential effects of berberine on influenza virus infection both in vitro and in vivo.Methods：In vitro anti-influenza virus assays were performed by cytopathogenic effect and neuraminidase assays in Madin Darby canine kidney cells.In vivo anti-influenza virus assays were performed on the viral pneumonia model of mice.The numbers of mice that died within day 2 to day 14 postinfection were recorded to calculate the mortality.On days 2,4,and 6,the viral titers in the lungs were determined by hemagglutination assay;hematoxylin/eosin staining was used to assess the pathogenic changes of lung tissues; the concentrations of tumor necrosis factor-alpha（TNF-α） and monocyte specific chemoattractant molecule （MCP-1） were measured by radio immunoassay or enzyme-linked immunosorbent assay;the concentrations of nitric oxide（NO） and inducible nitric oxide synthetase（iNOS） were detected by colorimetric method;reverse transcription polymerase chain reaction was used to detect the mRNA level of TNF-αand MCP-1.Results： Berberine showed inhibitory effects on cytopathogenic effects and neuraminidase activity of virus,with the therapeutic index 9.69.In vivo,berberine decreased mice mortality from 90%to 55%,reduced virus titers in the lungs on day 2 postinfection（P0.05）.The lung histology scores were 1.50±0.67,4.50±1.00,and 5.50±1.00 in the berberine group on days 2,4,and 6,respectively,which were significantly reduced compared to 2.17±0.22,6.83±0.44,and 8.50±0.33 in the infected group（P0.05）.The productions of NO and iNOS were repressed by berberine compared with those in the infected group（P0.01）.The transcription and expression of TNF-αwere inhibited by berberine on day 4（P0.01） and day 6（P0.05）,and those of MCP-1 were inhibited on day 6（P0.01） compared with the infected group.Conclusions：Berberine exhibited antiviral effects on the influenza virus both in vitro and in vivo.The possible therapeutic mechanism of berberine on influenza-induced viral pneumonia might be inhibiting the virus infection,as well as improving the pathogenic changes by repressing inflammatory substances release

In Vitro and In Vivo Antibacterial Activity of Patchouli Alcohol from Pogostemon cablin

Objective:To investigate the antibacterial activity of patchouli alcohol(PA)against 127 bacteria strains,including the common bacteria and drug-resistant bacteria strains both in the in vitro and in vivo tests.Methods:For the in vitro trial,the antibacterial property of PA against 107 Gram-positive and 20 Gram-negative bacteria strains was screened by agar double dilution method.For the in vivo trial,specific pathogen free Kunming strain of both male and female white mice,were used to test the protective ability of PA after being injected with the median lethal dose of the tested strains.Results:PA possessed antibacterial activity against all the tested 127 strains.In the in vitro test,PA could inhibit both Gram-negative bacteria(25-768μg/m L)and Gram-positive bacteria(1.5-200μg/m L).Particularly,PA was active against some drug-resistant bacteria like methicillin-resistant Staphylococcus aureus(MRSA).PA also exhibited in vivo anti-MRSA activity in mice via intraperitoneal injection.PA could protect mice entirely infected with MRSA at 100 and 200 mg/kg,while 80% mice injected with MRSA could be protected at a low dose of 50 mg/kg.Conclusion:PA might be a potential antibacterial drug from natural sources and might be worthy to explore its mechanism and application in further study.

Roxithromycin的药效学研究

报道国产ｒｏｘｉｔｈｒｏｍｙｃｉｎ（ＲＯＸ）对２０５株临床致病菌体内外抗菌活性及不同试验条件对抗菌活性的影响，并与进口产品及红霉素、麦迪霉素、柱晶白霉素、青霉素钾盐的抗菌活性进行比较。还进行了杀菌试验，结果表明，国产与进口ＲＯＸ及红霉素体外抗菌作用相似，对革兰氏阳性和阴性球菌有较好的抑制作用。另外，ＲＯＹ在偏碱性环境中抑菌效果较强，对１０５、１０７ｃｆｕ／ｍｌ细菌的ＭＩＣ基本一样。对感染金葡球菌的小鼠有很好的治疗保护作用，优于红霉素。

ANTIVIRAL EFFECTS ON MOUSE LEUKEMIA VIRUS REPLICATION BY OLIGODEOXYNUCLEOTIDES IN VITRO AND IN VIVO

Oligodeoxynucleotides (Oligomers) including modified and unmodified, homo-and heterooligomers were tested for their ability to inhibit mouse SRS leukemia virus (SRSV)-induced proliferation of cells, colony formation, syncytium formation and reverse transcriptase (RT) activity in vitro. Phosphorothioate analogs complementary to Mo-MuLV sequences, as well as noncomplementary homooligomers, were found to be active. Unmodified homooligomer (dC14) also showed inhibition of growth of ascitic lymphoma carrying SRS virus. Our study suggests that different classes of oligonucleotides may inhibit SRSV replication with different mechanisms.

Antioxidant Activities of Crude Phlorotannins from Sargassum hemiphyllum

Brown algae are well known as a source of biologically active compounds, especially those having antioxidant activities, such as phlorotannins. In this study we examined the antioxidant activities of crude phlorotannins extracts（CPEs） obtained from Sargassum hemiphyllum（SH） and fractionated according to the molecular weights. When CPEs were administrated at a dose of 30 mg/kg to Kunming mice pre-treated with carbon tetrachloride（CCl4）, the levels of oxidative stress indicators in the liver, kidney and brain were significantly reduced in vivo. All the components of various molecular weight fractions of CPEs exhibited greater scavenging capacities in clearing hydroxyl free radical and superoxide anion than the positive controls gallic acid, vitamin C and vitamin E. Particularly, the components greater than 30 k D obtained from ethyl acetate phase showed the highest antioxidant capacities. These results indicated that SH is a potential source for extracting phlorotannins, the algal antioxidant compounds.

射干止咳胶囊体内外抗甲型H1N1流感病毒作用初探

目的初步探讨射干止咳胶囊对甲型H1N1流感病毒的体内外抗病毒作用。方法通过细胞病变效应(cytopathic effect,CPE)结合MTT法考察射干止咳胶囊体外抑制甲型H1N1流感病毒活性的作用;采用小鼠滴鼻法感染甲型H1N1流感病毒为模型,观察射干止咳胶囊对甲型H1N1流感病毒的体内抗病毒作用。结果体外抗病毒试验结果显示,射干止咳胶囊对甲型H1N1流感病毒的最高抑制率为11.86%,提示在攻毒剂量为100TCID_(50)的条件下,射干止咳胶囊在体外对甲型H1N1流感病毒的抑制作用不明显。小鼠滴鼻感染5LD_(50)的甲型H1N1流感病毒,射干止咳胶囊低、中、高剂量组小鼠的死亡率分别为30%、10%、30%,死亡保护率达50%、70%、50%,平均生存天数分别为13 d、14 d、13 d,说明各给药组对攻毒小鼠具有较好的死亡保护作用。结论射干止咳胶囊体外抗甲型H1N1流感病毒的作用不显著,但射干止咳胶囊各剂量组对甲型H1N1流感病毒攻毒致小鼠死亡却具有明显的保护作用。

Anthelmintic, antimicrobial, antioxidant and cytotoxic activity of Caltha palustris var. alba Kashmir, India

The methanolic extract obtained from the root portion of Caltha palustris var. alba was evaluated for its anthelmintic efficacy against gastrointestinal nematodes of sheep under both in vitro and in vivo conditions using worm motility inhibition(WMI) assay and fecal egg count reduction(FECR) assay, respectively. The extract was subjected to antimicrobial activity using agar-well diffusion method against different bacterial strains. In addition the extract was evaluated for cytotoxic and antioxidant activity against cultured THP-1(Leukemia), A-549(Lung), HCT-15(Colon), Cervix(HeLa) and PC-3(Prostrate) cell lines by SRB and DPPH radical scavenging assays. The extract used resulted in mean %WMI of 94.44%, as observed when the worms were put in lukewarm buffer for 30 min after exposure to different treatments. The mean mortality index of the sample was 0.95. The lethal concentration(LC50) was 0.11 mg·mL-1. Cell lines were exposed to concentration of 100 μg·mL-1 of extract for 48 h, which reduced the viability of these cell lines. The same plant extract also showed 55.58% DPPH radical scavenging activity.

黄秋葵抗氧化活性研究

目的研究黄秋葵果实不同极性部位提取物的体外和体内抗氧化能力,为黄秋葵的产品开发和利用提供依据。方法采用·DPPH清除能力、·ABTS^(+)清除能力、·OH清除能力、·O_(2)^(-)清除能力、FRAP法还原能力、TBARS法抗脂质过氧化能力体外抗氧化模型对黄秋葵果实95%乙醇提取物中不同极性部位进行抗氧化活性评价,并利用SPSS 19.0软件分析黄秋葵果实抗氧化活性与总黄酮含量之间的相关性。选择其具有体外抗氧化活性部位的提取物进行体内抗氧化活性研究。结果95%乙醇提取物中乙酸乙酯层和正丁醇层提取物表现出清除自由基能力、还原能力和抑制脂质过氧化能力,乙酸乙酯层和正丁醇层提取物的总黄酮含量高于其他各层提取物,总黄酮含量与抗氧化活性呈较强相关性。与模型组比较,给予乙酸乙酯层和正丁醇层提取物能够升高CCl_(4)肝损伤大鼠肝脏中SOD、CAT活性以及GSH水平,同时能够降低ALT活性和MDA水平。结论黄秋葵果实具有潜在的护肝作用,可为开展肝脏损伤保护研究提供重要依据。

百里香酚体外抗伪狂犬病毒活性评价及其作用方式

采用细胞体外培养技术,以BHK-21细胞为研究模型,探讨不同质量浓度的百里香酚在体外抗伪狂犬病毒(PRV)的活性及其作用方式。使用CCK-8法检测百里香酚对BHK-21细胞的最大无害浓度(MNTC),计算得出其半数抑制浓度(IC50)为(212.789±1.652)μg·mL^(-1),对PRV的半数有效浓度(EC50)为(30.710±0.303)μg·mL^(-1),对PRV的治疗指数(TI)为6.929,说明百里香酚属于高效低毒类抗PRV药物。使用CPE观察法检测百里香酚对PRV病毒滴度和病毒一步生长曲线的影响,结果显示,百里香酚能够剂量依赖性显著降低PRV的病毒滴度,并降低PRV在BHK-21中增殖后的毒力。采用荧光定量PCR方法检测不同处理下PRV感染的BHK-21细胞的PRV-gE基因的拷贝量,研究百里香酚体外抗PRV的作用方式,结果显示,百里香酚主要是通过抑制PRV在BHK-21细胞中的增殖来发挥抗病毒活性的,同时能够直接杀灭部分PRV病毒粒子。

瑞香狼毒总黄酮提取物的抗肿瘤作用

目的: 研究瑞香狼毒总黄酮提取物的抗肿瘤作用。方法:体外抗肿瘤活性研究采用MTT比色法和集落形成法测定,靶细胞采用人胃癌细胞SGC- 790 1、人肝癌细胞BEL- 74 0 2和人白血病细胞HL- 6 0 ;通过提取物对小鼠的移植性肿瘤S180和H2 2生长的影响来评价体内抗肿瘤作用。结果:瑞香狼毒总黄酮提取物对体外培养的肿瘤细胞有较强的抑制作用,体外抗肿瘤活性高于长春新碱;瑞香狼毒总黄酮提取物急性毒性较小,对小鼠移植性肿瘤S180和H2 2的生长也有显著的抑制作用,并和剂量正相关,高剂量组对小鼠移植性肿瘤S180的抑制率为4 5 . 6 4 % ,H2 2的为4 7 .5 9% ,与阳性对照组环磷酰胺的抑制率接近。结论:瑞香狼毒总黄酮提取物具有较强的体内和体外抗肿瘤作用。

藤茶多糖的抗氧化作用研究

目的: 探讨藤茶多糖的体内外抗氧化作用。 方法: (1)测定藤茶多糖(Ampelopsis Grossedentata polysaccharide,AGP)的还原能力;(2)测定AGP在化学模拟体系、体外血清的抗活性氧能力;(3)测定AGP对小鼠肝匀浆、小鼠肝线粒体MDA生成的影响;(4)用分光光度法测定H2O2诱导小鼠红细胞溶血和Fe2+-VC诱导的肝线粒体肿胀程度来研究AGP的抗氧化效果;(5)小鼠随机分成对照与AGP组,腹腔注射AGP 150mg/kg·bw·d 12d后,测定小鼠血清抗活性氧能力及肝组织MDA含量。 结果: AGP具有一定的还原能力,一定浓度范围内具有显著的体外抗活性氧能力并能抑制小鼠肝组织及肝线粒体MDA的生成,可减少红细胞诱导溶血和肝线粒体肿胀程度,对小鼠体内肝组织MDA生成的抑制作用显著。 结论: AGP一定浓度范围内具有抗氧化作用。

猴耳环抗病毒有效成分研究

目的：对抗炎中药猴耳环的化学成分进行研究。方法：采用体外抗病毒活性追踪的方法，应用大孔吸附树脂吸附，聚酰胺、硅胶和凝胶柱色谱技术进行分离，根据理化性质和波谱数据鉴定其结构。结果：分离并鉴定了6个化合物：tricetiflavan（5，7，3′，4′，5′-pentahydroxylflavan）（1），myricitrin（myricetin-3-O-α-L-rhamnopyranoside）（2），quercitrin（quercetin-3-O-α-L-rhamnopyranoside）（3），quercetin（4），methyl gallate（5）和gallic acid（6）。结论：化合物1-5为首次从陔植物中分离得到，其中化合物4被发现有较好的体外抗呼吸道合胞病毒（RSV）活性。

芦苇叶黄酮类提取物体内体外抗氧化性研究

研究醇提芦苇叶黄酮类提取物的体内体外抗氧化性。通过测定芦苇叶黄酮类提取物对体外大鼠肝组织匀浆、肝线粒体丙二醛(malondialdehyde,MDA)生成量及大鼠红细胞溶血的影响。结果表明:芦苇叶黄酮类提取物在0.5~10 mg/m L范围内能抑制大鼠肝组织匀浆及肝线粒体MDA的生成,抑制大鼠红细胞溶血,具有量效关系,表明芦苇叶黄酮类提取物具有体外抗氧化效果。随着剂量的增加,芦苇叶黄酮类提取物可提高S180荷瘤小鼠血清中谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)和超氧化物歧化酶(superoxide dismutase,SOD)活性,降低MDA含量,从而提高其体内抗氧化活性,当剂量达到300 mg/(kg·d)时,芦苇叶黄酮类提取物与10 m L/(kg·d)的氟尿嘧啶均可促进荷瘤小鼠体内抗氧化能力的提高,且二者间没有显著性差异。

桃胶多糖体内外抗氧化作用的研究

为了研究桃胶多糖体内外抗氧化的作用,实验采用体外测定桃胶多糖对1,1-二苯基-2-三硝基苯肼自由基(DPPH·)、2,2'-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐阳离子自由基(ABTS^+·)、羟基自由基(·OH)和超氧阴离子(O-2·)等的清除率及对铁的还原能力,体内通过建立D-半乳糖小鼠氧化损伤模型,给予不同剂量(2、4、6 g/kg)桃胶多糖灌胃,测定血清及肝脏中丙二醛(MDA)含量及总抗氧化能力(T-AOC)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性。结果显示,桃胶多糖体外对DPPH·、ABTS^+·、·OH和O2^-·的半数抑制浓度(IC50)分别为6.95、0.56、1.10、0.11 mg/mL,且对铁还原力随着浓度(0.02~0.14 mg/mL)的升高而增大;在体内实验中,与模型组相比,连续服用不同剂量(2、4、6 g/kg)的桃胶多糖能够显著或极显著降低小鼠血清及肝脏中MDA含量、提高T-AOC能力、增强SOD及GSH-Px的活性(P<0.05或P<0.01),同时剂量间存在一定的量效关系。研究结果表明桃胶多糖具有良好的体内外抗氧化活性,可进一步申报成为新型的食品资源,应用于制药与食品工业中抗氧化、抗衰老、降血糖等保健产品的开发。