目的:对从FDA(food and drug administration)批准的1430个化合物中通过微复制子系统筛选出对发热伴血小板减少综合征病毒(severe fever with thrombocytopenia syndrome virus,SFTSV)起抑制作用的药物,并解析药物抗病毒的作用阶段。方...目的:对从FDA(food and drug administration)批准的1430个化合物中通过微复制子系统筛选出对发热伴血小板减少综合征病毒(severe fever with thrombocytopenia syndrome virus,SFTSV)起抑制作用的药物,并解析药物抗病毒的作用阶段。方法:利用SFTSV微复制子初步筛选出对SFTSV复制转录系统具有抑制作用的药物,通过药物浓度梯度抑制实验确定各药物的半数抑制浓度(IC50)。再利用SFTSV感染细胞模型,使用药物分别与病毒孵育后再感染细胞、与病毒共同和细胞孵育、在病毒与细胞孵育后作用于细胞、以及在病毒感染细胞全过程(entire stage)使用药物,并采用实时荧光定量PCR(qRT-PCR)对感染细胞上清中病毒进行定量,分析药物对病毒感染细胞的整个过程的不同阶段,包括:病毒颗粒稳定性(virion stability)、病毒感染入侵(entry stage)、病毒进入细胞后的复制过程(post-entry)的抑制作用,并与药物作用于病毒感染细胞整个过程的抑制作用相比较,初步揭示药物抑制SFTSV感染的作用机制。结果:吗替麦考酚酯、麦考酚酸、硝唑尼特、Vidofludimus 4种药物对SFTSV具有较好的抑制效果,4种药物对微复制子系统的半数抑制浓度IC50分别为0.014、0.627、1.283、0.059μmol/L;4种药物对SFTSV的抑制作用发生在病毒入侵细胞后的阶段。结论:吗替麦考酚酯、麦考酚酸、硝唑尼特、Vidofludimus具有较好的抗发热伴血小板减少综合征病毒效果。展开更多
Chemical reactions and fate of the toxins of Bacillus thuringiensis （Bt） in the soil environment are causing increasing concerns due to the large-scale cultivation of transgenic Bt plants. In this study, the effect ...Chemical reactions and fate of the toxins of Bacillus thuringiensis （Bt） in the soil environment are causing increasing concerns due to the large-scale cultivation of transgenic Bt plants. In this study, the effect of ionic strength （0-1 000 mmol kg-1） adjusted by NaCl or CaCl2 on adsorption of Bt toxin by a lateritic red soil, a paddy soil and these soils after chemical removal of organic-bound or free Fe and Al oxides, as well as by pure minerals （goethite, hematite and gibbsite） which are widespread in these soils, were studied. The results indicated that when the supporting electrolyte was NaCl, the adsorption of Bt toxin by the lateritic red soil and paddy soil increased rapidly until the ionic strength reached 250 mmol kg-1 and then gradually slowed down with the increase of ionic strength; while in ease the supporting electrolyte was CaCl2, the adsorption of Bt toxin enhanced significantly at low ionic strength （〈 10 mmol kg-1） and then decreased as the ionic strength increased. The adsorption of Bt toxin by the tested minerals and soils after the removal of organic-bound or free Fe and Al oxides also increased with increasing ionic strength controlled by NaCl. Removing organic-bound Fe and Al oxides obviously increased the adsorption of Bt toxin in the tested soils. Differently, removing free Fe and Al oxides increased the Bt adsorption by the paddy soil, but decreased the adsorption by the lateritic red soil. The study indicated that the varieties of ionic strength and the presence of Ve and Al oxides affected the adsorption of Bt toxin by the soils, which would contribute to the further understanding of the fate of Bt toxin in the soil environment and provide references for the ecological risk assessment of transgenic Bt plants.展开更多
Objective:To screen the anti-SFTSV drugs from 1430 FDA-approved drugs via mini-genome system,and to investigate which stage of the infection process could be suppressed by the identified drugs.Methods:The SFTSV mini-g...Objective:To screen the anti-SFTSV drugs from 1430 FDA-approved drugs via mini-genome system,and to investigate which stage of the infection process could be suppressed by the identified drugs.Methods:The SFTSV mini-genome system was used to screen drugs with inhibitory effect on SFTSV replication and transcription,and the 50%inhibitory concentration(IC_(50))of each drug was calculated by drug concentration gradient inhibition experiment.Drugs were used to pre-incubate with virus and then incubate with cells,to incubate with virus and cells simultaneously,to incubate with cells after virus invading into cells,or to incubate during the whole infection process,and then qRT-PCR was used to measure the viral RNA copies in the culture supernatant.These experiments were performed to quantitatively determine the inhibition effects of drugs on SFTSV indifferent stages of the whole process including virion stability,entry and post-entry stages,so as to clarify the inhibition mechanism of these drugs.Results:Four drugs including Mycophenolate mofetil,Mycophenolic acid,Nitazoxanide,and Vidofludimus were identified having efficient inhibitory effects on SFTSV RNA replication via minigenome system,with the IC_(50) of 0.014μmol/L,0.627μmol/L,1.283μmol/L,and 0.059μmol/L,respectively.All four drugs showed effective inhibition when adding during the whole SFTSV infection process as well as the post-entry stage.Conclusion:Mycophenolate mofetil,Mycophenolic acid,Nitazoxanide and Vidofludimus show efficient anti-viral effects on SFTSV infection.展开更多
文摘目的:对从FDA(food and drug administration)批准的1430个化合物中通过微复制子系统筛选出对发热伴血小板减少综合征病毒(severe fever with thrombocytopenia syndrome virus,SFTSV)起抑制作用的药物,并解析药物抗病毒的作用阶段。方法:利用SFTSV微复制子初步筛选出对SFTSV复制转录系统具有抑制作用的药物,通过药物浓度梯度抑制实验确定各药物的半数抑制浓度(IC50)。再利用SFTSV感染细胞模型,使用药物分别与病毒孵育后再感染细胞、与病毒共同和细胞孵育、在病毒与细胞孵育后作用于细胞、以及在病毒感染细胞全过程(entire stage)使用药物,并采用实时荧光定量PCR(qRT-PCR)对感染细胞上清中病毒进行定量,分析药物对病毒感染细胞的整个过程的不同阶段,包括:病毒颗粒稳定性(virion stability)、病毒感染入侵(entry stage)、病毒进入细胞后的复制过程(post-entry)的抑制作用,并与药物作用于病毒感染细胞整个过程的抑制作用相比较,初步揭示药物抑制SFTSV感染的作用机制。结果:吗替麦考酚酯、麦考酚酸、硝唑尼特、Vidofludimus 4种药物对SFTSV具有较好的抑制效果,4种药物对微复制子系统的半数抑制浓度IC50分别为0.014、0.627、1.283、0.059μmol/L;4种药物对SFTSV的抑制作用发生在病毒入侵细胞后的阶段。结论:吗替麦考酚酯、麦考酚酸、硝唑尼特、Vidofludimus具有较好的抗发热伴血小板减少综合征病毒效果。
基金Supported by the National Natural Science Foundation of China (Nos. 41001140 and 40671087)
文摘Chemical reactions and fate of the toxins of Bacillus thuringiensis （Bt） in the soil environment are causing increasing concerns due to the large-scale cultivation of transgenic Bt plants. In this study, the effect of ionic strength （0-1 000 mmol kg-1） adjusted by NaCl or CaCl2 on adsorption of Bt toxin by a lateritic red soil, a paddy soil and these soils after chemical removal of organic-bound or free Fe and Al oxides, as well as by pure minerals （goethite, hematite and gibbsite） which are widespread in these soils, were studied. The results indicated that when the supporting electrolyte was NaCl, the adsorption of Bt toxin by the lateritic red soil and paddy soil increased rapidly until the ionic strength reached 250 mmol kg-1 and then gradually slowed down with the increase of ionic strength; while in ease the supporting electrolyte was CaCl2, the adsorption of Bt toxin enhanced significantly at low ionic strength （〈 10 mmol kg-1） and then decreased as the ionic strength increased. The adsorption of Bt toxin by the tested minerals and soils after the removal of organic-bound or free Fe and Al oxides also increased with increasing ionic strength controlled by NaCl. Removing organic-bound Fe and Al oxides obviously increased the adsorption of Bt toxin in the tested soils. Differently, removing free Fe and Al oxides increased the Bt adsorption by the paddy soil, but decreased the adsorption by the lateritic red soil. The study indicated that the varieties of ionic strength and the presence of Ve and Al oxides affected the adsorption of Bt toxin by the soils, which would contribute to the further understanding of the fate of Bt toxin in the soil environment and provide references for the ecological risk assessment of transgenic Bt plants.
基金supported by Key Program of the Joint Fund of the National Natural Science Foundation of China(No.U20A20135)。
文摘Objective:To screen the anti-SFTSV drugs from 1430 FDA-approved drugs via mini-genome system,and to investigate which stage of the infection process could be suppressed by the identified drugs.Methods:The SFTSV mini-genome system was used to screen drugs with inhibitory effect on SFTSV replication and transcription,and the 50%inhibitory concentration(IC_(50))of each drug was calculated by drug concentration gradient inhibition experiment.Drugs were used to pre-incubate with virus and then incubate with cells,to incubate with virus and cells simultaneously,to incubate with cells after virus invading into cells,or to incubate during the whole infection process,and then qRT-PCR was used to measure the viral RNA copies in the culture supernatant.These experiments were performed to quantitatively determine the inhibition effects of drugs on SFTSV indifferent stages of the whole process including virion stability,entry and post-entry stages,so as to clarify the inhibition mechanism of these drugs.Results:Four drugs including Mycophenolate mofetil,Mycophenolic acid,Nitazoxanide,and Vidofludimus were identified having efficient inhibitory effects on SFTSV RNA replication via minigenome system,with the IC_(50) of 0.014μmol/L,0.627μmol/L,1.283μmol/L,and 0.059μmol/L,respectively.All four drugs showed effective inhibition when adding during the whole SFTSV infection process as well as the post-entry stage.Conclusion:Mycophenolate mofetil,Mycophenolic acid,Nitazoxanide and Vidofludimus show efficient anti-viral effects on SFTSV infection.
