苯基吡啶酮衍生物影响猪δ冠状病毒复制的作用分析

本研究旨在探讨苯基吡啶酮衍生物JIB-04对猪δ冠状病毒（porcine deltacoronavirus,PDCoV）的抗病毒作用,并初步研究可能的作用机制。首先应用CCK-8方法检测不同浓度JIB-04作用后的细胞活力,并计算半数毒性浓度（CC₅₀）和半数有效浓度（EC₅₀）。应用TCID₅₀方法检测JIB-04预处理和共处理对PDCoV复制的影响,检测JIB-04处理对PDCoV吸附或入侵细胞的影响。最后,应用qRT-PCR、TCID₅₀和Western blot方法检测JIB-04处理对不同时间点病毒复制的影响。结果表明,JIB-04在所有受试浓度下均不影响LLC-PK细胞的活力,CC₅₀>640 μmol·L^-1,EC₅₀=0.216 μmol·L^-1,SI指数>2 963。与未处理的病毒感染组相比,JIB-04处理极显著降低了PDCoV的滴度（P<0.001）,但对PDCoV吸附和入侵过程没有影响。感染6 h后,与未处理的病毒感染组相比,JIB-04处理组PDCoV滴度极显著下降（P<0.01）;感染12和24 h后,PDCoV滴度、基因组拷贝数、N蛋白表达水平均极显著下降（P<0.01）。JIB-04的细胞毒性较低,药物选择性指数较高,在体外能抵抗PDCoV感染,可以作为潜在的抗病毒药物。在PDCoV感染过程中,JIB-04能够抑制病毒核酸和蛋白质的合成,抑制病毒的复制。     

商陆抗病毒蛋白在植物抗病上的应用

商陆抗病毒蛋白是指商陆属植物中的核糖体失活蛋白.阐述了商陆抗病毒蛋白的研究历史、意义、种类和理化性质,对病原菌的抗性、抗病机理及其在植物抗病上的应用情况等,并对它的应用前景作了展望.     

MicroRNA研究概述

MicroRNA(miRNA)是一类真核生物内源性非编码的单链小分子RNA,长约21nt~23nt。越来越多的miRNA在动物细胞和植物组织中发现,这些成熟的小分子RNA是从具有发夹结构的前体miRNA(pre-miRNA)剪切出来,通过与靶mRNA分子互补结合而抑制蛋白翻译或导致mRNA降解,从而调控靶基因的表达。miRNA是一类重要的基因调控子,在机体的基因表达调控、细胞分化和凋亡以及抗病毒防御中发挥重要的作用。深入理解miRNA介导的宿主与病毒之间的相互调节作用,对于阐明病毒的致病机理与制定治疗策略具有深远的意义。     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.     

Aurasperone A Inhibits SARS CoV-2 In Vitro: An Integrated In Vitro and In Silico Study

Several natural products recovered from a marine-derived Aspergillus niger were tested for their inhibitory activity against SARS CoV-2 in vitro. Aurasperone A (3) was found to inhibit SARS CoV-2 efficiently (IC₅₀ = 12.25 µM) with comparable activity with the positive control remdesivir (IC₅₀ = 10.11 µM). Aurasperone A exerted minimal cytotoxicity on Vero E6 cells (CC₅₀ = 32.36 mM, SI = 2641.5) and it was found to be much safer than remdesivir (CC₅₀ = 415.22 µM, SI = 41.07). To putatively highlight its molecular target, aurasperone A was subjected to molecular docking against several key-viral protein targets followed by a series of molecular dynamics-based in silico experiments that suggested M^pro to be its primary viral protein target. More potent anti-SARS CoV-2 M^pro inhibitors can be developed according to our findings presented in the present investigation.     

鸡突变型和未突变型Mx蛋白体外抗病毒活性研究

【目的】研究鸡Mx蛋白第631位氨基酸变异与抗病性的相关性。【方法】用已经构建成功的中国狼山鸡Mx蛋白基因突变型pcDNA3.0-MMx和未突变型pcDNA3.0-Mx真核表达载体分别转染鸡成纤维（CEF）细胞和鼠成纤维（NIH-3T3）细胞;利用RT-PCR检测突变型MMx基因和未突变型Mx基因的表达,微量细胞病变抑制法测定重组蛋白抗新城疫病毒（NDV）和水泡性口炎病毒（VSV）的效果。【结果】诱变鸡Mx基因的表达重组蛋白可保护CEF细胞在孵育48 h内免受NDV的感染;转染突变型pcDNA3.0-MMx真核表达载体的NIH-3T3细胞在孵育60 h内亦未受到VSV的浸染;而转染未突变型pcDNA3.0-Mx真核表达载体的CEF和NIH-3T3细胞在24 h内均发生了病变。【结论】体外重组突变的Mx蛋白在单细胞水平上具有延缓NDV和VSV感染的能力。     

In Vitro Anti-Orthohantavirus Activity of the High-and Low-Molecular-Weight Fractions of Fucoidan from the Brown Alga Fucus evanescens

The Hantaan orthohantavirus (genovariant Amur–AMRV) is a rodent-borne zoonotic virus; it is the causative agent of haemorrhagic fever with renal syndrome in humans. The currently limited therapeutic options require the development of effective anti-orthohantavirus drugs. The ability of native fucoidan from Fucus evanescens (FeF) and its enzymatically prepared high-molecular-weight (FeHMP) and low-molecular-weight (FeLMP) fractions to inhibit different stages of AMRV infection in Vero cells was studied. The structures of derivatives obtained were determined using nuclear magnetic resonance (NMR) spectroscopy. We found that fucoidan and its derivatives exhibited significant antiviral activity by affecting the early stages of the AMRV lifecycle, notably virus attachment and penetration. The FeHMP and FeLMP fractions showed the highest anti-adsorption activity by inhibiting AMRV focus formation, with a selective index (SI) > 110; FeF had an SI of ~70. The FeLMP fraction showed a greater virucidal effect compared with FeF and the FeHMP fraction. It was shown by molecular docking that 2O-sulphated fucotetrasaccharide, a main component of the FeLMP fraction, is able to bind with the AMRV envelope glycoproteins Gn/Gc and with integrin β3 to prevent virus–cell interactions. The relatively small size of these sites of interactions explains the higher anti-AMRV activity of the FeLMP fraction.     

三黄鸡a干扰素的克隆、表达及抗病毒活性初步研究

采用PCR方法从三黄鸡血液基因组中扩增鸡α干扰素全基因,并克隆和测序.序列分析表明,基因全长为582 bp,亚克隆其成熟蛋白编码基因489 bp.将该片段与表达载体pET-28a连接克隆至大肠埃希菌DH5α菌株,经测序和酶切鉴定,选取正向插入、读码框正确的阳性克隆.构建重组质粒并转化BL21(DE3),经IPTG诱导,...     

伪狂犬病流行现状及我国应采取的防制措施

目的研究山柰提取物在体外抑制伪狂犬病病毒活性。方法通过响应面法优化山柰的超声提取工艺,并用荧光定量PCR法检测山柰提取物抗病毒的作用方式及其作用时间点。结果以黄酮含量作为响应值,优化得到最佳制备工艺：溶剂为80%乙醇,功率208 W,提取98 min;抗伪狂犬病毒活性研究表明：山柰提取物对伪狂犬病毒的半数抑制质量浓度(IC₅₀)为55.85 μg/mL,作用方式为抑制病毒的穿入和复制阶段,在病毒感染24 h内加入山柰提取物均表现出显著的抗病毒活性。结论山柰提取物具有显著的体外抗伪狂犬病毒活性。