为探究des(rhamnosyl)verbascoside体外抗乙型肝炎病毒(HBV)的活性作用和机制,本研究以des(rhamnosyl)verbascoside为实验药物对HepG2.2.15细胞进行干预,实验分为药物干预组和对照组,采用串联质谱标签(Tandem Mass Tag,TMT)蛋白质组学...为探究des(rhamnosyl)verbascoside体外抗乙型肝炎病毒(HBV)的活性作用和机制,本研究以des(rhamnosyl)verbascoside为实验药物对HepG2.2.15细胞进行干预,实验分为药物干预组和对照组,采用串联质谱标签(Tandem Mass Tag,TMT)蛋白质组学方法对提取的总蛋白进行分析。结果表明,共筛选得到300个差异表达蛋白,其中有109个上调蛋白,191个下调蛋白。基因本体论(Gene Ontology,GO)分析结果显示,差异蛋白主要参与DNA复制(DNA replication)、鞘糖脂代谢(Glycosphingolipid metabolic process)、细胞增殖(Cell proliferation)、寡糖分解代谢(Oligosaccharide catabolic process)等生物学过程,以及DNA聚合酶活性(DNA polymerase activity)、丝氨酸型羧肽酶活性(Serine type carboxypeptidase activity)、DNA引物酶活性(DNA primase activity)等分子功能。京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析结果显示,差异蛋白主要参与细胞代谢(Metabolism)、遗传信息传导(Genetic information processing)、生物系统通路(Organismal systems)等相关信号通路。亚细胞定位分析表明,差异蛋白大多定位在细胞质和细胞核。本研究共筛选出13个与抗HBV密切相关的蛋白。通过定量蛋白组学初步揭示des(rhamnosyl)verbascoside可能通过增加HGF、SORT1、MAN2B1,减少PRIM1、PRIM2、POLA1、POLD3、POLD2、POLD1、POLE、ERCC2、LAMC1、SDC1等蛋白表达来起到体外抗HBV的作用。展开更多
Discrete event system(DES)models promote system engineering,including system design,verification,and assessment.The advancement in manufacturing technology has endowed us to fabricate complex industrial systems.Conseq...Discrete event system(DES)models promote system engineering,including system design,verification,and assessment.The advancement in manufacturing technology has endowed us to fabricate complex industrial systems.Consequently,the adoption of advanced modeling methodologies adept at handling complexity and scalability is imperative.Moreover,industrial systems are no longer quiescent,thus the intelligent operations of the systems should be dynamically specified in the model.In this paper,the composition of the subsystem behaviors is studied to generate the complexity and scalability of the global system model,and a Boolean semantic specifying algorithm is proposed for generating dynamic intelligent operations in the model.In traditional modeling approaches,the change or addition of specifications always necessitates the complete resubmission of the system model,a resource-consuming and error-prone process.Compared with traditional approaches,our approach has three remarkable advantages:(i)an established Boolean semantic can be fitful for all kinds of systems;(ii)there is no need to resubmit the system model whenever there is a change or addition of the operations;(iii)multiple specifying tasks can be easily achieved by continuously adding a new semantic.Thus,this general modeling approach has wide potential for future complex and intelligent industrial systems.展开更多
[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PP...[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.展开更多
[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were ...[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E.coli.Furthermore,based on the purified soluble N protein and NH fusion protein,a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established.[Results]The method could quickly and quantitatively detect PPR V antibody in sheep serum,with high sensitivity and specificity,without any cross reaction to other related sheep pathogens.The intra-batch and inter-batch coefficients of variation were less than 10%and 15%,respectively,and the method had good repeatability.Through detection on 292 clinical serum samples,it was compared with the French IDVET competitive ELISA kit,and the coincidence rate of the two methods reached 93.84%.Compared with the serum neutralization test,the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum(provided by the WOAH Brucellosis Reference Laboratory of France).[Conclusions]This method can realize rapid quantitative detection of PPR V antibody on site,and has high practical value and popularization value.展开更多
文摘为探究des(rhamnosyl)verbascoside体外抗乙型肝炎病毒(HBV)的活性作用和机制,本研究以des(rhamnosyl)verbascoside为实验药物对HepG2.2.15细胞进行干预,实验分为药物干预组和对照组,采用串联质谱标签(Tandem Mass Tag,TMT)蛋白质组学方法对提取的总蛋白进行分析。结果表明,共筛选得到300个差异表达蛋白,其中有109个上调蛋白,191个下调蛋白。基因本体论(Gene Ontology,GO)分析结果显示,差异蛋白主要参与DNA复制(DNA replication)、鞘糖脂代谢(Glycosphingolipid metabolic process)、细胞增殖(Cell proliferation)、寡糖分解代谢(Oligosaccharide catabolic process)等生物学过程,以及DNA聚合酶活性(DNA polymerase activity)、丝氨酸型羧肽酶活性(Serine type carboxypeptidase activity)、DNA引物酶活性(DNA primase activity)等分子功能。京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)分析结果显示,差异蛋白主要参与细胞代谢(Metabolism)、遗传信息传导(Genetic information processing)、生物系统通路(Organismal systems)等相关信号通路。亚细胞定位分析表明,差异蛋白大多定位在细胞质和细胞核。本研究共筛选出13个与抗HBV密切相关的蛋白。通过定量蛋白组学初步揭示des(rhamnosyl)verbascoside可能通过增加HGF、SORT1、MAN2B1,减少PRIM1、PRIM2、POLA1、POLD3、POLD2、POLD1、POLE、ERCC2、LAMC1、SDC1等蛋白表达来起到体外抗HBV的作用。
基金supported by the National Natural Science Foundation of China(U21B2074,52105070).
文摘Discrete event system(DES)models promote system engineering,including system design,verification,and assessment.The advancement in manufacturing technology has endowed us to fabricate complex industrial systems.Consequently,the adoption of advanced modeling methodologies adept at handling complexity and scalability is imperative.Moreover,industrial systems are no longer quiescent,thus the intelligent operations of the systems should be dynamically specified in the model.In this paper,the composition of the subsystem behaviors is studied to generate the complexity and scalability of the global system model,and a Boolean semantic specifying algorithm is proposed for generating dynamic intelligent operations in the model.In traditional modeling approaches,the change or addition of specifications always necessitates the complete resubmission of the system model,a resource-consuming and error-prone process.Compared with traditional approaches,our approach has three remarkable advantages:(i)an established Boolean semantic can be fitful for all kinds of systems;(ii)there is no need to resubmit the system model whenever there is a change or addition of the operations;(iii)multiple specifying tasks can be easily achieved by continuously adding a new semantic.Thus,this general modeling approach has wide potential for future complex and intelligent industrial systems.
基金Supported by National Key R&D Program for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39)+2 种基金Key Research and Development Program of Shandong Province(Major Science and Technology Innovation Project)(2021CXGC011306)Scientific Research Project of General Administration of Customs(2024HK033)Scientific Research Project of Jinan Customs(2023JK005).
文摘[Objectives]This study was conducted to explore rapid and large-scale screening and detection of peste des petits ruminants(PPR),so as to provide important technical means for prevention,control and purification of PPR.[Methods]Soluble N protein and NH fusion protein were successfully obtained in an Escherichia coli expression system by optimizing E.coli codon and expression conditions.Furthermore,based on purified soluble N protein and NH fusion protein,a double-antigen sandwich time-resolved fluorescence immunoassay method for detection of peste des petits ruminants virus(PPRV)was established.[Results]The method has high sensitivity and specificity and can specifically detect the antibody against PPRV in sheep serum,and it has no cross reaction with other related diseases.The method was used to detect 292 clinical samples,and compared with French IDVET competition ELISA kit.The coincidence rates of positive samples and negative samples from the two kinds of test kits were 92.47%and 97.26%,respectively,and the overall coincidence rate was 94.86%.The intra-group and inter-group coefficients of variation in the repeatability test were less than 10%.[Conclusions]Compared with the traditional ELISA method,the double-antigen sandwich time-resolved fluorescence immunoassay for detection of PPRV has equivalent sensitivity and specificity,and simple and rapid operation,and thus high application and popularization value.
基金Supported by The National Project for the Prevention and Control of Major Exotic Animal Diseases(2022YFD1800500)National Mutton Sheep Industrial Technology System(CARS39).
文摘[Objectives]This study was conducted to establish a rapid quantitative method for detecting antibody against Peste des Petits Ruminants Virus(PPR V)in sheep serum.[Methods]Soluble N protein and NH fusion protein were obtained in Escherichia coli prokaryotic expression system by optimizing codons and expression conditions of E.coli.Furthermore,based on the purified soluble N protein and NH fusion protein,a high-sensitivity fluorescence immunoassay kit for detecting the antibody against PPR V was established.[Results]The method could quickly and quantitatively detect PPR V antibody in sheep serum,with high sensitivity and specificity,without any cross reaction to other related sheep pathogens.The intra-batch and inter-batch coefficients of variation were less than 10%and 15%,respectively,and the method had good repeatability.Through detection on 292 clinical serum samples,it was compared with the French IDVET competitive ELISA kit,and the coincidence rate of the two methods reached 93.84%.Compared with the serum neutralization test,the detected titer value of the high-sensitivity rapid fluorescence quantitative detection method was basically consistent with the tilter value obtained by the neutralization test on the standard positive serum(provided by the WOAH Brucellosis Reference Laboratory of France).[Conclusions]This method can realize rapid quantitative detection of PPR V antibody on site,and has high practical value and popularization value.