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SARS-CoV-2靶向CypA/CD147受体途径诱导心肌细胞凋亡
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作者 马明仁 马凌 +1 位作者 刘燕 王菲 《西安交通大学学报(医学版)》 CAS CSCD 北大核心 2024年第5期734-740,共7页
目的探究SARS-CoV-2靶向亲环素A(CypA)/细胞外金属蛋白酶诱导剂(CD147)受体途径诱导心肌细胞凋亡的可能机制。方法构建pEncmv-SARS-CoV-2 S-3xflag重组载体质粒转染H9c2细胞,TUNEL实验、流式细胞术和DNA ladder分析细胞凋亡,免疫共沉淀(... 目的探究SARS-CoV-2靶向亲环素A(CypA)/细胞外金属蛋白酶诱导剂(CD147)受体途径诱导心肌细胞凋亡的可能机制。方法构建pEncmv-SARS-CoV-2 S-3xflag重组载体质粒转染H9c2细胞,TUNEL实验、流式细胞术和DNA ladder分析细胞凋亡,免疫共沉淀(CoIP)验证CD147与SARS-CoV-2 S蛋白之间相互作用,Western blotting法检测细胞凋亡信号通路相关蛋白表达。结果与对照组比较,TUNEL实验和流式细胞术结果显示转染组细胞凋亡显著增加(P<0.05),DNA ladder分析发现DNA降解明显。Western blotting法检测显示Caspase12、DR3表达显著升高(P<0.001),FAS表达升高(P<0.01),TRF1、DR4表达降低(P<0.01)。结论SARS-CoV-2 S蛋白靶向CypA/CD147受体入侵宿主细胞,激活内质网通路(Caspase12)和死亡受体通路(DR3、FAS),以内源性和外源性凋亡途径诱导心肌细胞凋亡。 展开更多
关键词 新型冠状病毒(sars-cov-2) 新型冠状病毒肺炎(COVID-19) 亲环素A(CypA)/细胞外金属蛋白酶诱导剂(CD147) 心肌细胞 信号通路 凋亡
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SARS-COV-2 M^(pro)抑制剂2-(2-氯苯基)-7-(异喹啉-4-基)-5,7-二氮杂螺[3.4]辛-6,8-二酮的合成
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作者 闫翱翔 臧瑞涵 +2 位作者 李华 陈丽霞 李行舟 《沈阳药科大学学报》 CAS CSCD 2024年第9期1198-1204,共7页
目的研究SARS-COV-2 M^(pro)抑制剂2-(2-氯苯基)-7-(异喹啉-4-基)-5,7-二氮杂螺[3.4]辛-6,8-二酮(1)的合成方法,为其深入研究和其衍生物的合成提供借鉴方法。方法以(2-氯苯基)乙酸甲酯为起始原料,经亲电取代、还原、磺酰化、环化、选择... 目的研究SARS-COV-2 M^(pro)抑制剂2-(2-氯苯基)-7-(异喹啉-4-基)-5,7-二氮杂螺[3.4]辛-6,8-二酮(1)的合成方法,为其深入研究和其衍生物的合成提供借鉴方法。方法以(2-氯苯基)乙酸甲酯为起始原料,经亲电取代、还原、磺酰化、环化、选择性水解得到3-(2-氯苯基)-1-乙氧羰基环丁烷-1-甲酸(6),在三乙胺存在下,化合物6与叠氮磷酸二苯酯反应,生成的酰基叠氮化合物经Curtius重排生成相应的异氰酸酯;再与4-氨基异喹啉反应,并在碳酸钠的作用下进一步环合生成目标化合物。结果中间体及目标化合物的化学结构经^(1)H-NMR、^(13)C-NMR、ESI-MS确证。结论对化合物6的合成路线进行了优化;通过化合物6获得了一条更精简的化合物1的合成路线,该路线既避免了有毒的三光气的使用,也避免了副反应的发生;通过制备型HPLC得到化合物1的两个顺反异构体,并使用NOESY确定了化合物的构型。 展开更多
关键词 严重急性呼吸窘迫综合征冠状病毒2型 主蛋白酶 抑制剂 合成方法
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靶向SARS-CoV-2的纳米抗体研究进展
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作者 程小龙 孟妮 +1 位作者 刘喜富 陈静 《河北师范大学学报(自然科学版)》 CAS 2024年第3期294-302,共9页
席卷全球的新型冠状病毒病(corona virus disease 2019,COVID-19)是由严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)引起的,SARS-CoV-2是继严重急性呼吸综合征冠状病毒(severe acute respi... 席卷全球的新型冠状病毒病(corona virus disease 2019,COVID-19)是由严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)引起的,SARS-CoV-2是继严重急性呼吸综合征冠状病毒(severe acute respiratory syndrome coronavirus,SARS-CoV)和中东呼吸综合征冠状病毒(middle east respiratory syndrome coronavirus,MERS-CoV)后人类发现的第三种引起全球大流行的冠状病毒.刺突蛋白(spike protein,S)的受体结合域(receptor binding domain,RBD)可以与细胞表面的血管紧张素转换酶2(angiotensin converting enzyme2,ACE2)结合,进而入侵细胞内部,启动病毒的复制.为了预防和治疗新冠病毒肺炎,研究人员研发了多种疫苗、小分子药物及抗体药物.纳米抗体(nanobodies,Nbs)是可识别抗原的最小结合片段,具有体积小、渗透性高、热稳定性好、结合特异性高、生产成本低、免疫原性小等优势,在治疗SARS-CoV-2中效果显著.对SARS-CoV-2的纳米抗体研究进展进行了综述,这些纳米抗体的发现,对于新冠病毒病的诊断和治疗均有潜在的应用价值. 展开更多
关键词 冠状病毒 新型冠状病毒 刺突蛋白 纳米抗体
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一种吡唑并吡啶酮类衍生物对新型冠状病毒SARS-CoV-2的抑制作用研究
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作者 李银燕 姜琳瑞 +3 位作者 陈至烜 梁锦龙 何晓雪 杨洁 《沈阳药科大学学报》 CAS CSCD 2024年第7期914-921,共8页
目的探究3-(4-氯苯基)-1,4-二苯基-1,4,5,7-四氢-6H-吡唑并[3,4-b]吡啶-6-酮(以下简称为J1)体外抑制新型冠状病毒(SARS-CoV-2)的作用及潜在作用机制。方法采用MTT法评价化合物J1对Vero-E6、ACE2/293T和HRT18细胞的毒性。利用感染性SARS-... 目的探究3-(4-氯苯基)-1,4-二苯基-1,4,5,7-四氢-6H-吡唑并[3,4-b]吡啶-6-酮(以下简称为J1)体外抑制新型冠状病毒(SARS-CoV-2)的作用及潜在作用机制。方法采用MTT法评价化合物J1对Vero-E6、ACE2/293T和HRT18细胞的毒性。利用感染性SARS-CoV-2和人冠状病毒OC43(HCoV-OC43)检测化合物J1的抗冠状病毒活性。采用SARS-CoV-2假病毒体外细胞感染模型和时间点加药实验检测J1抑制SARS-CoV-2病毒感染靶细胞的作用阶段。通过S蛋白介导的细胞融合抑制实验检测J1是否通过阻止病毒膜融合而抑制SARS-CoV-2的进入。采用SPR实验和分子对接技术阐明J1与SARS-CoV-2 S蛋白的作用。结果J1对Vero-E6、ACE2/293T和HRT18细胞的毒性较小,半数细胞毒性浓度CC50均大于100μmol·L^(-1)。J1能显著抑制SARS-CoV-2原始株和Delta变异株的活性,半数有效浓度EC50分别为607μmol·L^(-1)和221μmol·L^(-1)。此外,J1可抑制HCoV-OC43病毒的感染,显著减少NP蛋白和mRNA的表达。分子对接结果显示,J1通过氢键作用和疏水作用力与SARS-CoV-2 S蛋白活性位点稳定结合。机制研究结果表明,J1可抑制SARS-CoV-2病毒进入靶细胞,半数抑制浓度IC50为157μmol·L^(-1)。J1浓度依赖性抑制SARS-CoV-2 S蛋白介导的细胞-细胞膜融合,SPR实验证实J1与S蛋白具有较强结合能力。结论吡唑并吡啶酮类衍生物J1通过靶向S蛋白抑制SARS-CoV-2进入靶细胞。 展开更多
关键词 sars-cov-2 S 蛋白 抗冠状病毒药物 小分子化合物 进入抑制剂
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Huu S. TIEU’s Predicting Outcome of Severe Acute Respiratory Syndrome (SARS) and Preparing the Treatment for COVID-19 (Coronavirus) and Other Viral Pandemics
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作者 Huu S. Tieu Martin F. Loeffler 《International Journal of Clinical Medicine》 CAS 2023年第2期110-115,共6页
This project paper is to give a prediction for the future of other viral pandemics and to provide recommendations for preparing therapies that could help in the success of effective treatments and benefits for patient... This project paper is to give a prediction for the future of other viral pandemics and to provide recommendations for preparing therapies that could help in the success of effective treatments and benefits for patients in life-threatening situations. The theory of prediction was proposed by Huu S. TIEU on March 25, 2019, and he hypothesized that any malfunctioning cell in the body could have a damaging effect. This paper discusses the prediction that Localized Oxygen Deprivation could be a contributing factor for a future epidemic or other viral pandemics that could affect body function. This paper is based on opinion and does not have sufficient evidence to support the claims made. Therefore, further in-depth study is needed to prove the findings. The author cites Hypoxia to support his idea, but he is not claiming that Hypoxia-Inducible Factor (HIF) has worked on his predictions. The author also tested a theory using cow blood curd for body function, but this test was not a structured test and the findings were not supported by other evidence. To further prove the idea or theory, further study into the subject should be conducted. 展开更多
关键词 COVID-19 (coronavirus) Severe Acute Respiratory Syndrome (sars) Viral Pandemics Huu S. TIEU Life Is Quantum Biology
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Natural Products as Potential Leads Against Coronaviruses: Could They be Encouraging Structural Models Against SARS-CoV-2? 被引量:1
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作者 Ilkay Erdogan Orhan F.Sezer Senol Deniz 《Natural Products and Bioprospecting》 CAS 2020年第4期171-186,共16页
New coronavirus referred to SARS-CoV-2 has caused a worldwide pandemic(COVID-19)declared by WHO.Coronavirus disease 2019(COVID-19)is an infectious disease with severe acute respiratory syndrome caused by coronavirus-2... New coronavirus referred to SARS-CoV-2 has caused a worldwide pandemic(COVID-19)declared by WHO.Coronavirus disease 2019(COVID-19)is an infectious disease with severe acute respiratory syndrome caused by coronavirus-2(SARS-CoV-2).SARS-CoV-2 is akin to SARS-CoV,which was the causative agent of severe acute respiratory syndrome(SARS)in 2002 as well as to that of Middle East respiratory syndrome(MERS)in 2012.SARS-CoV-2 has been revealed to belong to Coronaviridiae family as a member ofβ-coronaviruses.It has a positive-sense single-stranded RNA with the largest RNA genome.Since its genomic sequence has a notable similarity to that of SARS-CoV,antiviral drugs used to treat SARS and MERS are now being also applied for COVID-19 treatment.In order to combat SARS-CoV-2,many drug and vaccine development studies at experimental and clinical levels are currently conducted worldwide.In this sense,medicinal plants and the pure natural molecules isolated from plants have been reported to exhibit significant inhibitory antiviral activity against SARS-CoV and other types of coronaviruses.In the present review,plant extracts and natural molecules with the mentioned activity are discussed in order to give inspiration to researchers to take these molecules into consideration against SARS-CoV-2. 展开更多
关键词 coronavirus sars-cov Medicinal plants Natural products ANTIVIRAL
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Diabetes and coronavirus infections (SARS-CoV, MERS-CoV, and SARS-CoV-2)
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作者 Azadeh Haghi Navand Saber Soltani +3 位作者 Mona Moghadami Parastoo Hosseini SepidehNasimzadeh Milad Zandi 《Journal of Acute Disease》 2020年第6期244-247,共4页
Diabetes,as the major cause of hyperglycemia,is a chronic metabolic disease.Immune system disorders caused by diabetes can increase the risk of respiratory diseases.Thus,diabetes is considered to be a major risk facto... Diabetes,as the major cause of hyperglycemia,is a chronic metabolic disease.Immune system disorders caused by diabetes can increase the risk of respiratory diseases.Thus,diabetes is considered to be a major risk factor for viral respiratory infections such as coronavirus infections.Coronaviruses are members of the Coronaviridae,which has caused three outbreaks from 2003 to 2020.Patients with coronavirus infection in the lower and upper respiratory tract could show mild to severe symptoms.In this review,we focus on the relationship between diabetes and three coronaviruses:SARS-CoV,MERS-CoV,and SARS-CoV-2. 展开更多
关键词 DIABETES coronavirus sars-cov MERS-CoV sars-cov-2
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基于分子对接和分子动力学模拟技术筛选SARS-CoV-23CL蛋白酶抑制剂 被引量:3
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作者 吉冰洁 李尚校 +3 位作者 王俭 边远 刘浩 赵勇山 《沈阳药科大学学报》 CAS CSCD 北大核心 2023年第8期1069-1077,1091,共10页
目的 以分子对接和分子动力学模拟技术相结合的方法筛选得到严重急性呼吸系统综合征冠状病毒2 (severe acute respiratory syndrome coronavirus 2,SARS-CoV-2) 3CL蛋白酶的小分子抑制剂。方法 基于SARS-CoV-2 3CL蛋白酶的晶体结构(PDB ... 目的 以分子对接和分子动力学模拟技术相结合的方法筛选得到严重急性呼吸系统综合征冠状病毒2 (severe acute respiratory syndrome coronavirus 2,SARS-CoV-2) 3CL蛋白酶的小分子抑制剂。方法 基于SARS-CoV-2 3CL蛋白酶的晶体结构(PDB ID:7K3T和7SI9),从PubChem数据库中筛选得到SARS-CoV-2 3CL蛋白酶的特异性抑制剂nirmatrelvir的结构类似物,使用Autodock进行非共价分子对接,使用AutoDockFR进行共价分子对接,采用Pymol和Ligplot软件对SARS-CoV-2 3CL蛋白酶和小分子进行相互作用模式分析,最后使用AMBER18进行小分子化合物与SARS-CoV-2 3CL蛋白酶复合物的分子动力学模拟,进一步验证分子对接结果,使用SwissADME对化合物进行成药性分析。结果 将78个nirmatrelvir结构类似物与SARS-CoV-2 3CL蛋白酶进行虚拟筛选和分子对接,得到了2个与nirmatrelvir作用相当的小分子化合物(PubChem ID:57842182和156619968),共价对接发现化合物57842182和化合物156619968与SARS-CoV-2 3CL蛋白酶间不存在共价结合。分子动力学模拟结果表明化合物57842182、化合物156619968与SARS-CoV-2 3CL蛋白酶的结合自由能(ΔE_(total))分别为-23.96 kcal·mol^(-1)和-27.11 kcal·mol^(-1),二者主要通过与MET165、GLU166、ASN142、CYS145、LEU50、MET49等氨基酸残基的相互作用占据SARS-CoV-2 3CL蛋白酶亚活性位点,具有潜在抑制SARS-CoV-2 3CL蛋白酶活性的功能。化合物57842182和化合物156619968的成药性略差于nirmatrelvir。结论 筛选得到的nirmatrelvir结构类似物57842182和156619968具有作为新型冠状病毒SARS-CoV-2 3CL蛋白酶小分子抑制剂的潜力,其与SARS-CoV-2 3CL蛋白酶的结合特点可为SARS-CoV-2 3CL蛋白酶抑制剂的药物设计提供线索。 展开更多
关键词 新型冠状病毒 sars-cov-23CL蛋白酶 分子对接 分子动力学模拟
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Site discrepancy of synonymous codon usage in SARS coronavirus and other viruses in Coronaviridae
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作者 周童 顾万君 +2 位作者 马建民 孙啸 陆祖宏 《Journal of Southeast University(English Edition)》 EI CAS 2005年第2期203-206,共4页
The synonymous codon usage in the translational initiation and termination regions of genes of severe acute respiratory syndrome (SARS) coronavirus and five other viruses in Coronaviridae was systematically analyzed.T... The synonymous codon usage in the translational initiation and termination regions of genes of severe acute respiratory syndrome (SARS) coronavirus and five other viruses in Coronaviridae was systematically analyzed.The results indicate that most minor codons for these coronaviruses are preferentially used in the initial and terminal region.The minor codons preferentially used in the initial region are thought to have a negative effect on gene expression,which can be explained by the minor codon modulator hypothesis.It also indicates that the minor codons preferentially used in the terminal region may regulate the level of gene expression.The proposed results strongly imply that the minor codon modulator hypothesis can be applied to both some bacteria and some viruses. 展开更多
关键词 codon usage severe acute respiratory syndrome (sars) coronavirus gene expression site discrepancy
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Bat SARS coronavirus HKU3-1病毒株基因组及蛋白质的生物信息学分析
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作者 李钧 张阳德 +2 位作者 胡智渊 彭锴 潘一峰 《中国医学工程》 2007年第8期625-627,632,共4页
目的比较中国菊头蝠SARS冠状病毒(Bat SARS coronavirus)与人类及果子狸体内分离出来的SARS-Cov,分析它们是否存在一定的相似性。方法本文分析了Bat SARS coronavirus与已知冠状病毒Human SARS-CoV和civet SARS-CoV的系统进化关系,绘制... 目的比较中国菊头蝠SARS冠状病毒(Bat SARS coronavirus)与人类及果子狸体内分离出来的SARS-Cov,分析它们是否存在一定的相似性。方法本文分析了Bat SARS coronavirus与已知冠状病毒Human SARS-CoV和civet SARS-CoV的系统进化关系,绘制了Bat SARS coronavirusHKU3-1的全基因序列图,同时预测了S蛋白和N蛋白的三维模型。结果对Bat SARS coronavirus HKU3-1的序列分析表明中国菊头蝠SARS-CoV与人和果子狸SARS-CoV非常相近。系统进化分析法显示Bat-SARS-CoV与SARS-CoV形成一个不同的群,称为group 2b CoV,与已知的group 2 CoV存在一定的距离。结论Bat SARS coronavirus HKU3-1不太可能是从人SARS-CoV传播过来的。另外,Bat SARS coronavirus与civet SARS-CoV可能具有共同的祖先,使得中国菊头蝠SARS病毒株同样具有感染人体的危险。 展开更多
关键词 中国菊头蝠sars冠状病毒 系统进化分析法 基因序列图
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Impacts of SARS-CoV-2 on diabetes mellitus:A pre and post pandemic evaluation
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作者 A H M Nurun Nabi Akio Ebihara Hossain Uddin Shekhar 《World Journal of Virology》 2023年第3期151-171,共21页
The coronavirus disease 2019(COVID-19)pandemic caused by the novel beta coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)crippled the whole world and has resulted in large number of morbidity and... The coronavirus disease 2019(COVID-19)pandemic caused by the novel beta coronavirus severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)crippled the whole world and has resulted in large number of morbidity and mortality.The origin of the SARS-CoV-2 is still disputed.The risk of infection with SARS-CoV-2 is dependent on several risk factors as observed in many studies.The severity of the disease depends on many factors including the viral strain,host immunogenetics,environmental factors,host genetics,host nutritional status and presence of comorbidities like hypertension,diabetes,Chronic Obstructive Pulmonary Disease,cardiovascular disease,renal impairment.Diabetes is a metabolic disorder mainly characterized by hyperglycemia.Diabetic individuals are intrinsically prone to infections.SARS-CoV-2 infection in patients with diabetes result inβ-cell damage and cytokine storm.Damage to the cells impairs the equilibrium of glucose,leading to hyperglycemia.The ensuing cytokine storm causes insulin resistance,especially in the muscles and liver,which also causes a hyperglycemic state.All of these increase the severity of COVID-19.Genetics also play pivotal role in disease pathogenesis.This review article focuses from the probable sources of coronaviruses and SARS-CoV-2 to its impacts on individuals with diabetes and host genetics in pre-and post-pandemic era. 展开更多
关键词 coronavirus sars-cov-2 DIABETES MERS sars Single nucleotide polymorphism
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SARS coronavirus entry into host cells through a novel clathrin- and caveolae-independent endocytic pathway 被引量:51
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作者 Wang,H Yang,P +4 位作者 Liu,K Guo,F Zhang,Y Zhang,G Jiang,C 《Cell Research》 SCIE CAS CSCD 2008年第2期290-301,共12页
While severe acute respiratory syndrome coronavirus (SARS-CoV)~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endo... While severe acute respiratory syndrome coronavirus (SARS-CoV)~as initially thought to enter cells through direct fusion with the plasma membrane, more recent evidence suggests that yirus entry may also involve endocytosis. We have found that SARS-CoV enters cells viapH- and receptor-dependent endocytosis. Treatment of cells with either SARS-COV spike protein or spike-bearing pseudoviruses resulted in the translocation of angiotensin-converting enzyme 2 (ACE2), the functional receptor of SARS-CoV, from the cell surface to endosomes. In addition, the spike-bearing pseudoviruses and early endosome antigen 1 were found to colocalize in endosomes. Further analyses using specific endocytic path- way inhibitors and dominant-negative Epsl5 as well as caveolin-1 colocalization study suggested that virus entry was mediated by a clathrin- and caveolae-independent mechanism. Moreover, cholesterol- and sphingolipid-rich lipid raft microdomains in the plasma membrane, which have been shown to act as platforms for many physiological signaling pathways, were shown to be involved in virus entry. Endocytic entry of SARS-CoV may expand the cellular range of SARS-CoV infection, and our findings here contribute to the understanding of SARS-CoV pathogenesis, providing new information for anti-viral drug research. 展开更多
关键词 Severe Acute Respiratory Syndrome coronavirus sars-cov ENDOCYTOSIS angiotensin-converting enzyme 2 (ACE2) lipid rafts
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Immunogenicity of the Spike Glycoprotein of Bat SARS-like Coronavirus 被引量:1
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作者 Yu-xuan HOU Cheng PENG +3 位作者 Zheng-gang HAN Peng ZHOU Ji-guo CHEN Zheng-li SHI 《Virologica Sinica》 SCIE CAS CSCD 2010年第1期36-44,共9页
A group of SARS-like coronaviruses(SL-CoV)have been identified in horseshoe bats.Despite SL-CoVs and SARS-CoV share identical genome structure and high-level sequence similarity,SL-CoV does not bind to the same cellul... A group of SARS-like coronaviruses(SL-CoV)have been identified in horseshoe bats.Despite SL-CoVs and SARS-CoV share identical genome structure and high-level sequence similarity,SL-CoV does not bind to the same cellular receptor as for SARS-CoV and the N-terminus of the S proteins only share 64%amino acid identity,suggesting there are fundamental differences between these two groups of coronaviruses.To gain insight into the basis of this difference,we established a recombinant adenovirus system expressing the S protein from SL-CoV(rAd-Rp3-S)to investigate its immune characterization.Our results showed that immunized mice generated strong humoral immune responses against the SL-CoV S protein.Moreover,a strong cellular immune response demonstrated by elevated IFN-γand IL-6 levels was also observed in these mice.However,the induced antibody from these mice had weaker cross-reaction with the SARS-CoV S protein,and did not neutralize HIV pseudotyped with SARS-CoV S protein.These results demonstrated that the immunogenicity of the SL-CoV S protein is distinct from that of SARS-CoV,which may cause the immunological differences between human SARS-CoV and bat SL-CoV.Furthermore,the recombinant virus could serve as a potential vaccine candidate against bat SL-CoV infection. 展开更多
关键词 sars coronavirus sars-cov sars-like coronavirus (SL-CoV) Spike glycoprotein Humoral immune response Cellular immune response
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Multiple Sequence Alignment of the M Protein in SARS-Associated and Other Known Coronaviruses 被引量:1
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作者 史定华 周晖杰 +2 位作者 王斌宾 顾燕红 王翼飞 《Journal of Shanghai University(English Edition)》 CAS 2003年第2期118-123,共6页
In this paper, we report a multiple sequence alignment result on the basis of 10 amino acid sequences of the M protein, which come from different coronaviruses (4 SARS associated and 6 others known). The alignment mo... In this paper, we report a multiple sequence alignment result on the basis of 10 amino acid sequences of the M protein, which come from different coronaviruses (4 SARS associated and 6 others known). The alignment model was based on the profile HMM (Hidden Markov Model), and the model training was implemented through the SAHMM (Self Adapting Hidden Markov Model) software developed by the authors. 展开更多
关键词 sars (Severe Acute Respiratory Syndrome) coronavirus M (Membrane or Matrix) protein multiple sequence alignment profile HMM.
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Immune Responses and Histopathological Changes in Rabbits Immunized with Inactivated SARS Coronavirus
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作者 Chuan-hai ZHANG Xin-jian LIU +5 位作者 Yi-fei WANG Jia-hai LU Huan-ying ZHENG Sheng XIONG Mei-ying ZHANG Qiu-ying LIU 《中国病毒学》 CSCD 2007年第5期360-365,共6页
To evaluate the immunogenicity of inactivated SARS coronavirus(SARS-CoV),three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant,adjuvant,and normal saline respective... To evaluate the immunogenicity of inactivated SARS coronavirus(SARS-CoV),three groups of rabbits were immunized three times at 2-week intervals with inactivated vaccine + adjuvant,adjuvant,and normal saline respectively.Eight batchs of serum were sampled from the auricular vein at day 7 to day 51,and specific IgG antibody titers and neutralizing antibody titers were detected by indirect ELISA and micro-cytopathic effect neutralizing test.Antibody specificity was identified by proteinchip assay.Histopathological changes were detected by H&E staining.The results showed that,rabbits in the experimental group immunized with inactivated SARS-CoV all generated specific IgG antibodies with neutralizing activity,which suggested the inactivated SARS-CoV could preserve its antigenicity well and elicit an effective humoral immune responses.The peak titer value of specific IgG antibody and neutralizing antibody reached 1:40960 and 1:2560 respectively.In the experimental group,no obvious histopathological changes was detected in the H&E stained slides of heart,spleen,kidney and testis samples,but the livers had slight histopathological changes,and the lungs presented remarkable histopathological changes.These findings are of importance for SARS-CoV inactivated vaccine development. 展开更多
关键词 免疫反应 组织病理学 急性呼吸综合症 冠状病毒 sars 疫苗
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用SARS冠状病毒全基因组芯片杂交方法分析SARS-CoV 被引量:6
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作者 吴小兵 任鲁风 +8 位作者 马鑫 何新洲 袁振华 刘凤杰 赵革新 杨宇 张猛 董小岩 侯云德 《中国生物工程杂志》 CAS CSCD 2003年第7期89-93,97,共6页
为从临床样品中检测和分析SARS CoV病毒打基础 ,并为分析SARS CoV病毒的复制和转录等机理提供一种有效方法。以SARS冠状病毒TOR2株序列作为标准设计和制备一种覆盖SARS冠状病毒全基因组的寡聚核苷酸芯片 ,探针长度为 70nt,每相邻的探针... 为从临床样品中检测和分析SARS CoV病毒打基础 ,并为分析SARS CoV病毒的复制和转录等机理提供一种有效方法。以SARS冠状病毒TOR2株序列作为标准设计和制备一种覆盖SARS冠状病毒全基因组的寡聚核苷酸芯片 ,探针长度为 70nt,每相邻的探针序列重复 2 5nt,共660条。用该芯片分析了细胞培养的SARS CoV病毒总RNA、7个SARS CoV病毒的基因克隆片段。对RNA样品用随机引物进行反转录PCR获得cDNA。对DNA用随机引物扩增和dUTP cy3标记。结果用这种芯片杂交检测SARS CoV病毒RNA可见阳性信号呈全基因组分布 ,并且有多处连续的阳性信号点 ;用正常人的白细胞RNA为对照 ,杂交未出现明显阳性信号。检测 7个SARS CoV病毒基因克隆片段 ,在该片段相应的探针区段出现连续阳性信号点。这种方法可有效地检测和分析样品中SARS冠状病毒全基因组的信息。 展开更多
关键词 sars冠状病毒 全基因组 杂交方 sars-cov 基因芯片
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SARS-CoV及其他冠状病毒基因组比较分析 被引量:8
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作者 吴松锋 杜春娟 +6 位作者 万平 陈廷贵 荔建琦 李栋 朱云平 曾衍钧 贺福初 《遗传》 CAS CSCD 北大核心 2003年第4期373-382,共10页
对病毒种内和种间基因组的比较分析能获得很多关于病毒起源与演化的信息。对17株SARS-CoV的种内基因组变异分析发现共有137个变异位点,估算出SARS-CoV的突变率为8.04×10^(-3)核苷酸替换/位点/年。变异位点在基因组上的分布不均匀,... 对病毒种内和种间基因组的比较分析能获得很多关于病毒起源与演化的信息。对17株SARS-CoV的种内基因组变异分析发现共有137个变异位点,估算出SARS-CoV的突变率为8.04×10^(-3)核苷酸替换/位点/年。变异位点在基因组上的分布不均匀,变异位点最多的是基因组中编码S1蛋白的区域,而在编码依赖于RNA的RNA聚合酶区域中几乎没有变异位点。核苷酸和氨基酸替换的偏性预示变异可能不仅仅是由随机漂变产生。对冠状病毒种间基因组结构比较分析发现,SARS-CoV的基因组结构与IBV很相似;而保守基因系统发育分析表明,SARS-CoV属于冠状病毒的一个新分支,并且与血清型第二组冠状病毒进化关系较近。对其他某些分子特征的分析发现,在不同的方面SARS-CoV和不同组冠状病毒有不同的相似点。进一步对基因组非保守开放阅读框(ORF)的基序(motif)和跨膜区分析发现,各组冠状病毒基因组中位于基因S-E间的非保守ORF可能是同源的,但不是绝对必要的;而IBV和SARS-CoV的基因组中位于基因M-N间ORF可能不是同源的。综合分析SARS-CoV与3组血清型冠状病毒进化关系、宿主分布,以及SARS-CoV和IBV的s2m的进化关系,可以推测SARS-CoV有可能来自禽类。 展开更多
关键词 严重急性呼吸系统综合征 sars 冠状病毒 多态 进化
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SARS-CoVS蛋白全长基因克隆及其表达的初步研究 被引量:4
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作者 肖宇 倪兵 +5 位作者 李晋涛 李艳秋 黎万玲 王希良 姜曼 吴玉章 《中国临床康复》 CSCD 2003年第21期2932-2933,F003,共3页
目的:克隆表达SARS-CoVS蛋白,构建SARS全长基因疫苗。方法:从SARS病毒的总cDNA中以PCR方法扩增编码人SARS-CoVS1和S2蛋白的编码序列,再将二者分别克隆入真核表达载体pVAX1,构建重组表达载体。然后进一步将S1和S2连接到pVAX1中。酶切和... 目的:克隆表达SARS-CoVS蛋白,构建SARS全长基因疫苗。方法:从SARS病毒的总cDNA中以PCR方法扩增编码人SARS-CoVS1和S2蛋白的编码序列,再将二者分别克隆入真核表达载体pVAX1,构建重组表达载体。然后进一步将S1和S2连接到pVAX1中。酶切和测序鉴定阳性克隆。采用脂质体法转染VeroE6细胞,用Western检测S蛋白的表达。结果:PCR方法分别扩增出了1900bp和1800bp左右的基因片段,两片段插入pVAX1构建重组表达载体后,经序列测定证实该插入片段为SARS病毒S蛋白编码序列。将重组子转染VeroE6细胞,收集细胞总蛋白,Western检测获得特异蛋白带。结论:成功克隆并表达了SARS-CoVS蛋白,为其作为SARS基因疫苗研究打下基础。 展开更多
关键词 sars-covS蛋白 全长基因 克隆 sars病毒 基因疫苗 严重急性呼吸综合症
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原位杂交检测尸检组织中SARS-CoV RNA 被引量:11
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作者 张庆玲 丁彦青 +14 位作者 候金林 贺莉 黄仲曦 王慧君 蔡俊杰 张进华 张文丽 耿舰 李欣 康伟 杨磊 申洪 李祖国 韩惠霞 陆药丹 《第一军医大学学报》 CSCD 北大核心 2003年第11期1125-1127,共3页
目的从分子水平检测急性重症呼吸综合征(SARS)患者的病变组织中SARS病毒(SARS-associated coronavirus, SARS-CoV)的存在和分布情况。方法应用原位杂交技术检测因SARS死亡患者的肺、脾脏、淋巴结、垂体、胰腺、甲状旁腺、肾上腺、胃肠... 目的从分子水平检测急性重症呼吸综合征(SARS)患者的病变组织中SARS病毒(SARS-associated coronavirus, SARS-CoV)的存在和分布情况。方法应用原位杂交技术检测因SARS死亡患者的肺、脾脏、淋巴结、垂体、胰腺、甲状旁腺、肾上腺、胃肠道、皮肤、脑、肝、肾、血管、四肢横纹肌组织、骨髓、心脏、卵巢、子宫和睾丸等组织的SARS-CoV RNA的表达和定位。结果尸检组织多部位(包括肺泡上皮细胞、气管及支气管浆液腺上皮细胞、肺内单核/巨噬细胞、脾脏和淋巴结的单核/巨噬细胞、胰腺腺泡细胞、垂体嗜酸性细胞、肾上腺皮质细胞、甲状旁腺嗜酸性细胞、食道鳞状上皮、胃肠道上皮细胞及胃粘膜壁细胞、皮肤汗腺细胞、大脑神经元细胞、肝细胞、肾远曲小管上皮细胞、骨髓早幼粒细胞及小静脉内皮细胞)SARS-CoV RNA阳性。结论SARS-CoV可侵犯全身多种器官;SARS-CoV在机体的分布情况与冠状病毒受体CD13分布相似;皮肤汗腺、消化道上皮及肾远曲小管上皮细胞SARS-CoV RNA阳性对确定SARS传播途径具有重要意义。 展开更多
关键词 原位杂交 检测 尸检组织 sars-covRNA 急性重症呼吸综合征 sars
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SARS-CoVS蛋白功能性受体ACE2在人角膜、结膜中的表达 被引量:18
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作者 柳林 孙琰 +3 位作者 潘欣 沈炜 刘志勇 刘银平 《眼科研究》 CSCD 北大核心 2004年第6期561-564,共4页
目的检测人角膜、结膜中血管紧张素转化酶2(ACE 2)的表达,由此来初步推断重症急性呼吸综合征(SARS)冠状病毒由眼部入侵的可能.方法取成人眼球破裂伤摘除的眼角膜和结膜组织及4~6个月中期引产胎儿的眼角膜、结膜、心、肺组织,分别采用RT... 目的检测人角膜、结膜中血管紧张素转化酶2(ACE 2)的表达,由此来初步推断重症急性呼吸综合征(SARS)冠状病毒由眼部入侵的可能.方法取成人眼球破裂伤摘除的眼角膜和结膜组织及4~6个月中期引产胎儿的眼角膜、结膜、心、肺组织,分别采用RT-PCR和免疫组化方法检测组织中ACE 2的表达.取成人尸体解剖的心、肺组织作为对照.结果在上述各种组织中均检测到了ACE.2的表达.结论眼角膜、结膜是人体暴露于SARS冠状病毒的一个重要部位,研究结果从mRNA和蛋白质水平证实了眼部组织存在SARS冠状病毒S蛋白的功能性受体ACE 2,由此初步推断SARS-CoV存在由眼部入侵的可能,为临床进一步研究SARS的防护和致病机制提供了线索. 展开更多
关键词 sars-cov S蛋白 功能性受体 血管紧张素转化酶 角膜 结膜
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