Development and characterization of a clinical strain of Coxsackievirus A16 and an eGFP infectious clone

Coxsackievirus A16(CA16) is one of the major causes of hand, foot, and mouth disease(HFMD) worldwide, which is a common illness that affects children. The frequent occurrence of HFMD outbreaks has become a serious public health problem in Asia. Therefore, it is important to understand the pathogenesis and replication of CA16. In this study, a stable infectious c DNA clone of an epidemic strain of Coxsackievirus A16(CA16) was assembled, and subsequently a reporter virus(e GFP-CA16) was constructed by inserting the e GFP gene between the 5'-UTR and the N-terminus of VP4, with the addition of a 2A protease cleavage site(ITTLG) at its C-terminus. This was transfected into Vero cells to generate infectious recombinant viruses. The growth characteristics and plaque morphology, in vitro, in mammalian cells were found to be indistinguishable between the parental and recombinant viruses. Although the e GFP-CA16 showed smaller plaque size as compared to recombinant CA16, both were found to exhibit similar growth trends and EC50 of NITD008. In summary, this stable infectious c DNA clone should provide a valuable experimental system to study CA16 infection and host response. The e GFP-CA16 is expected to provide a powerful tool to monitor e GFP expression in infected cells and to evaluate the antiviral activity of potential antiviral agents in the treatment of CA16 infections.     

Anti-Cancer Activity of an Osthole Derivative,NBM-T-BMX-OS01: Targeting Vascular Endothelial Growth Factor Receptor Signaling and Angiogenesis

Angiogenesis occurs during tissue growth, development and wound healing. It is also required for tumor progression and represents a rational target for therapeutic intervention. NBM-T-BMX-OS01 (BMX), derived from the semisynthesis of osthole, an active ingredient isolated from Chinese herb Cnidium monnieri (L.) Cuss., was recently shown to enhance learning and memory in rats. In this study, we characterized the anti-angiogenic activities of NBM-T-BMX-OS01 (BMX) in an effort to develop novel inhibitors to suppress angiogenesis and tumor growth. BMX inhibited vascular endothelial growth factor (VEGF)-induced proliferation, migration and endothelial tube formation in human umbilical endothelial cells (HUVECs). BMX also attenuated VEGF-induced microvessel sprouting from aortic rings ex vivo and reduced HCT116 colorectal cancer cells-induced angiogenesis in vivo. Moreover, BMX inhibited the phosphorylation of VEGFR2, FAK, Akt and ERK in HUVECs exposed to VEGF. BMX was also shown to inhibit HCT116 cell proliferation and to suppress the growth of subcutaneous xenografts of HCT116 cells in vivo. Taken together, this study provides evidence that BMX modulates vascular endothelial cell remodeling and leads to the inhibition of tumor angiogenesis. These results also support the role of BMX as a potential drug candidate and warrant the clinical development in the treatment of cancer.     

Targeting of dicer-2 and RNA by a viral RNA silencing suppressor in Drosophila cells

RNA interference (RNAi) is a eukaryotic gene-silencing mechanism that functions in antiviral immunity in diverse organisms. To combat RNAi-mediated immunity, viruses encode viral suppressors of RNA silencing (VSRs) that target RNA and protein components in the RNAi machinery. Although the endonuclease Dicer plays key roles in RNAi immunity, little is known about how VSRs target Dicer. Here, we show that the B2 protein from Wuhan nodavirus (WhNV), the counterpart of Flock House virus (FHV), suppresses Drosophila melanogaster RNAi by directly interacting with Dicer-2 (Dcr-2) and sequestering double-stranded RNA (dsRNA) and small interfering RNA (siRNA). Further investigations reveal that WhNV B2 binds to the RNase III and Piwi-Argonaut-Zwille (PAZ) domains of Dcr-2 via its C-terminal region, thereby blocking the activities of Dcr-2 in processing dsRNA and incorporating siRNA into the RNA-induced silencing complex (RISC). Moreover, we uncover an interrelationship among diverse activities of WhNV B2, showing that RNA binding enhances the B2-Dcr-2 interaction by promoting B2 homodimerization. Taken together, our findings establish a model of suppression of Drosophila RNAi by WhNV B2 targeting both Dcr-2 and RNA and provide evidence that an interrelationship exists among diverse activities of VSRs to antagonize RNAi.     

安徽省2000-2009年不同类型植被的变化及其与气温、降水的关系

为了了解安徽省不同类型植被与气候环境因子的关系,利用2000-2009年的逐月MODIS/NDVI数据、安徽省80个气象站逐日气温、降水资料和安徽省土地覆盖数据,研究了安徽省近10年来植被指数的时空变化特征及其与气温、降水的相关性.结果表明:近10年来,安徽省各覆盖类型的植被指数变化趋势不一致,作物种植区和城镇的植被指数增加显著,其他覆盖类型的植被指数变化不显著;林地的平均植被指数最高,作物种植区次之,城镇最低;植被指数的月变化在作物种植区呈现双峰型,而其他覆盖类型呈现单峰型.安徽省月平均NDVI分别与月平均气温、月降水量呈显著线性正相关和非线性正相关,降水量对植被NDVI的作用存在一个阈值,但年际关系均呈弱正相关.植被指数与气温的偏相关系数林地最高,作物种植区最低,自然植被区的偏相关系数较人工植被区的大,植被指数与降水的偏相关系数恰好相反.安徽省淮北平原中部和江淮之间北部大部分地区(非水浇地)的植被是由气温和降水的共同作用驱动;淮北中部少数格点和少数水体格点的植被是单独由降水作用驱动;除一些水面的格点是非气象因子驱动外,其他大部分地区的植被是单独由气温作用驱动.     

沙粒病毒聚合酶C端的表达纯化与结晶条件筛选 *

拉沙病毒和马秋波病毒同属于沙粒病毒科,人类感染后病死率很高,对人类生命健康造成了严重威胁。沙粒病毒在基因组转录过程中存在与其他分节段负链RNA病毒相似的抢帽机制,位于病毒聚合酶N端的核酸内切酶结构域负责切割宿主mRNA,而其C端负责结合宿主mRNA帽子。目前对哺乳动物沙粒病毒聚合酶C端的特征知之甚少。为此构建了拉沙病毒和马秋波病毒聚合酶C端的重组表达载体,并利用E.coli原核系统进行表达和后续纯化,获得了不同聚体形式的C端蛋白,通过分析超速离心实验和负染电镜观察对其不同的聚体形式进行了表征。最后通过大量筛选结晶条件,得到了拉沙病毒聚合酶C端的晶体,为进一步研究其三维结构和功能机制奠定了基础。     

Cloning,characterization, and expression analysis of goat (Capra hircus) phospholipid hydroperoxide glutathione peroxidase (PHGPx)

Phospholipid hydroperoxide glutathione peroxidase (PHGPx), as a ubiquitous antioxidant enzyme in the glutathione peroxidases (GPx) family, plays multiple roles in organisms. However, there is very little information on PHGPx in goats (Capra hircus). In this study, a full-length cDNA was cloned and characterized from Taihang black goat testes. The 844 bp cDNA contains an open reading frame (ORF) of 597 bp. The goat PHGPx nucleotide sequence contains a selenocysteine (sec) codon TGA^244-246, two potential start codons ATG^20-22 and ATG^108-110, a polyadenylation signal AATAAA^813-818 and selenocysteine insertion sequence (SECIS) motif AUGA^688-691, UGA^729-731 and AAA^703-705. As a selenoprotein, the active-site motifs and GPx family signature motifs LAFPCNQF^101-108 and WNFEK^165-170 were also found. The order of PHGPx mRNA expression levels was: testes >> heart > brain > epididymis > kidney > liver > lung > spleen > muscle. Real-time PCR and immunohistochemistry results revealed similar expression differences in different age testes, with high expression levels during adolescence. Immunofluorescence results suggested that PHGPx mainly expressed in Leydig cells and spermatids in mature goat testes.     

Convergent evolution of SARS-CoV-2 in human and animals

Recently,many SARS-CoV-2 variants including 501Y.V1,501Y.V2 and 501Y.V3 were detected in different regions(Table S1)and drew great attention from all over the world.The 501Y.V1 was firstly isolated in the United Kingdom(UK)(Davies et al.,2020)and featured with 7 substitutions including N501Y as well as 3 deletions in S protein.This variant was identified to increase the viral transmissibility by 56％in comparison with the preexisting strains.Days after this report,another SARS-CoV-2 variant(501Y.V2)featured with N501Y,K417N and E484K substitutions in S protein was supposed to rapidly outcompete the preexisting strains(Tegally et al.,2020)in South Africa.Besides,the 501Y.V3 variant was initially detected in Brazil and has caused rapidly increased infections with SNPs N501Y,K417T and E484K.Of them,N501Y,K417N/T and E484K are of particular interest because the N501Y was shared in all three variants and the K417N/T and E484K were detected simultaneous appeared with N501Y in 501Y.V2 and 501Y.V3.