Heptaketides with antiviral activity from three endolichenic fungal strains Nigrospora sp., Alternaria sp. and Phialophora sp

Two new heptaketides, (+)-(2S,3S,4aS)-altenuene (1a) and (-)-(2S,3S,4aR)-isoaltenuene (2a), together with six known compounds, (-)-(2R,3R,4aR)-altenuene (1b), (+)-(2R,3R,4aS)-isoaltenuene (2b), 5'-methoxy-6-methyl-biphenyl-3,4,3'-triol (3), alternariol (4), alternariol-9-methyl ether (5), and 4-hydroxyalternariol-9-methyl ether (6) were isolated from the EtOAc extract of an endolichenic fungal strain Nigrospora sphaerica (No.83-1-1-2). Compounds 1a and 1b were separated from enantiomers 1 by chiral HPLC, and so were 2a and 2b from enantiomers 2. Interestingly, 1-6 were also obtained from other two endolichenic fungal strains Alternaria alternata (No.58-8-4-1) and Phialophora sp. (No.96-1-8-1). The structures of 1-6 were elucidated by means of MS, HR-MS, NMR, and X-ray diffraction. Furthermore, the absolute configurations of 1a-2b were determined by CD experiments and CD calculation. Of these compounds, 4 and 5 showed antiviral activity against herpes simplex virus (HSV) in vitro, with IC(50) values of 13.5 and 21.3 μM, and with selective index (SI) values of 26.5 and 17.1, respectively.     

Antiviral activity of Eupatorium adenophorum leaf extract against tobacco mosaic virus

Eupatorium adenophorum Spreng is a major invasive plant in China causing great economic losses. Considerable effort has been expended searching for new and innovative methods for integrative management of this species. In this study, an E. adenophorum extract was tested for antiviral activity against tobacco mosaic virus (TMV) using the local lesion assay method, and also characterized. The E. adenophorum leaf extract was able to strongly inhibit TMV infection, with electron microscopic observations indicating that the virus particles had been disaggregated. The active components in the extract are thermostable, stable at acidic pH, and largely not dialyzable. Tobacco seedlings treated with the extract exhibited significantly elevated superoxide dismutase, peroxidase, and polyphenol oxidase activities. Our results—the first reported observations of antiviral activity from leaf extracts of E. adenophorum — suggest that this species is a promising source of antiviral substances for practical use.     

从江香猪源干扰素γ植物表达载体构建及在生菜中瞬时表达

为构建从江香猪源干扰素γ(IFNγ)植物表达载体,通过农杆菌真空渗透法在新鲜生菜中实现瞬时表达。参考GenBank登录的猪IFNγ基因序列设计1对特异性IFNγ引物,采用RT-PCR方法从贵州从江香猪源肝脏组织扩增IFNγ基因序列,将其克隆至植物表达载体PBI121,转化根癌农杆菌LBA4404,筛选阳性菌株提取质粒,对重组质粒进行PCR扩增、双酶切及测序鉴定;携带重组质粒根癌农杆菌真空渗透法感染生菜,GUS染色检测感染的生菜叶片,ELISA检测感染生菜叶片中从江香猪源IFNγ蛋白,分析重组质粒在生菜中表达效果;采用细胞病变抑制实验检测生菜中从江香猪源IFNγ蛋白抗病毒活性。结果表明:本研究成功地从贵州从江香猪源肝脏组织扩增获得IFNγ基因序列并构建了从江香猪源IFNγ植物表达载体PBI121-IFNγ;GUS染色感染的生菜叶片可见蓝色物质;ELISA检测感染生菜叶片中从江香猪源IFNγ蛋白含量为169.397、170.657、170.37 pg/mL;生菜中表达从江香猪源IFNγ经4^-6稀释在Marc145细胞上仍能够抑制100 TCID50的PRRSV的致细胞病变作用。本实验成功构建从江香猪源IFNγ植物表达载体PBI121-IFNγ,生菜中瞬时表达蛋白具有免疫反应性和抗病毒活性,为从江香猪源IFNγ药用植物蛋白的研究开发提供参考依据。     

1-苯基-1,4-二氢-6-甲基-3-酰肼羰基-4-哒嗪酮及其生物活性

用两种方法合成了1-芳基-1，4-二氢-6-甲基-3-酰肼羰基-4-哒嗪酮(4) 。选择了较好的一种方法合成了一系列的目标化合物以期找到具有生物活性的新型化合物。生测结果表明，标题化合物对黄瓜子叶生根具有较高的促进生长作用，一些化合物对烟草花叶病毒（ＭＴＶ）和水稻纹枯病具有较高的抑制作用。     

中国野桑蚕抗病毒蛋白基因(Lipase)的克隆与活性鉴定

从中国野桑蚕幼虫中肠细胞克隆获得了抗家蚕核型多角体病毒BmNPV的脂肪酶基因(Lipase)cDNA(GenBank:AY945212)。该基因cDNA大小906 bp,编码301个氨基酸,蛋白质分子质量约为28.9 kD。进一步克隆了其全长基因组,结果表明该基因由2 147 bp组成,包含4个外显子和3个内含子。该基因在野桑蚕体内的表达具有组织特异性,仅限于野桑蚕中肠组织表达,且在幼虫龄中表达水平较高,而在幼虫眠期和熟蚕期几乎没有表达。通过基因体外表达获得的重组蛋白Lipase,能够有效抑制BmNPV病毒对家蚕的感染,说明该蛋白具有较强的抗BmNPV的生物学活性。     

10种中药成分对CEF的增殖和抵抗NDV感染的影响

分别将5种浓度的黄芪多糖(APS)、当归多糖(CAPS)、淫羊藿多糖(EPS)、板蓝根多糖(IRPS)、蜂胶多糖(PPS)、黄芪黄酮(AF)、淫羊藿黄酮(EF)、蜂胶黄酮(PF)、黄芪皂甙(AS)和人参皂甙(GS)10种中药成分加入到培养24h的鸡胚成纤维细胞(CEF)中，再培养12h后接种新城疫病毒(NDV)，于病毒接种后72h用中性红染料吸收法测定CEF活性以评价各中药成分对细胞增殖及其抵抗病毒感染的影响。结果表明，10种中药成分均能不同程度地促进细胞增殖和抵抗病毒感染，前者以AF、APS、IRPS、PF、和GS的作用较强，后者以GS和IRPS的作用较强，部分中药成分在两方面作用中各有所长，且有一定的量效关系。     

猪ESD真核表达质粒的构建及其抑制口蹄疫病毒复制的研究

[目的]探究猪ESD蛋白的抗病毒功能,通过构建猪ESD真核表达质粒,研究其是否能够抑制口蹄疫病毒在PK-15细胞中的复制。[方法]从PK-15细胞中提取总RNA,反转录为c DNA,PCR扩增出猪的ESD基因,构建到pc DNATM3.1/Myc-His A真核表达载体上。通过Western blotting和实时定量PCR检测ESD蛋白的表达情况及其抗病毒功能。[结果]获得了猪ESD真核表达载体,转染后能够正常表达。FMDV感染PK-15细胞后,ESD转录水平显著上调;过表达ESD蛋白显著抑制FMDV的复制;下调表达ESD蛋白显著促进FMDV的复制。[结论]猪ESD蛋白能够抑制FMDV在PK-15细胞中的复制。     

植物抗病毒基因工程研究进展及存在的问题

将近年来植物抗病毒基因工程的方法策略总结归纳为3个方面进行了综述,即:利用来源于病毒的基因、利用植物自身的抗病毒基因和采用多基因策略。并就当前存在的问题及其发展趋势进行了探讨。     

中药提取物对IBDV感染SPF鸡的防治效果

为了确定中药提取物对IBDV感染的防治效果,将150羽SPF雏鸡随机分为对照组、模型组和中药提取物高、中、低剂量组,中药提取物高、中、低剂量组分别在饮水中添加10.0、5.0、2.5 g/L的中药提取物,连续添加7 d,对照组和模型组不添加任何药物.给药后,除对照组外,其余组雏鸡分别通过滴鼻接种IBDV,攻毒后连续观察...