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
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
Background Although traumatic brain injury can lead to opening the blood-brain barrier and leaking of blood substances （including water） into brain tissue, few studies of brain antigens leaking into the blood and t...Background Although traumatic brain injury can lead to opening the blood-brain barrier and leaking of blood substances （including water） into brain tissue, few studies of brain antigens leaking into the blood and the pathways have been reported. Brain antigens result in damage to brain tissues by stimulating the immune system to produce anti-brain antibodies, but no treatment has been reported to reduce the production of anti-brain antibodies and protect the brain tissue. The aim of the study is to confirm the relationship between immune injury and arachnoid granulations following traumatic brain injury, and provide some new methods to inhibit the immune injury.Methods In part one, methylene blue was injected into the rabbits’ cisterna magna after traumatic brain injury, and concentrations of methylene blue and tumor necrosis factor （TNF）-α in blood were detected to determine the permeability of arachnoid granulations. In part two, umbilical cord mesenchymal stem cells and immature dendritic cells were injected into veins, and concentrations of interleukin 1 （IL-1）, IL-10, interferon （IFN）-γ, transforming growth factor （TGF）-β, anti-brain antibodies （ABAb）, and IL-12 were measured by ELISA on days 1, 3, 7, 14 and 21 after injury, and the numbers of leukocytes in the blood were counted. Twenty-one days after injury, expression of glutamate in brain tissue was determined by immunohistochemical staining, and neuronal degeneration was detected by H＆E staining. Results In part one, blood concentrations of methylene blue and TNF-α in the traumatic brain injury group were higher than in the control group （P 〈0.05）. Concentrations of methylene blue and TNF-α in the trauma cerebrospinal fluid （CSF） injected group were higher than in the control cerebrospinal fluid injected group （P 〈0.05）. In part two, concentrations of IL-1, IFN-γ, ABAb, IL-12, expression of glutamate （Glu）, neuronal degeneration and number of peripheral blood leukocytes were lower in the group with cell treatment compared to the control group. IL-10 and TGF-β were elevated compared to the control group. Conclusions Traumatic brain injury can lead to stronger arachnoid granulations （AGs） permeability; umbilical cord mesenchymal stem cells and immature dendritic cells can induce immune tolerance and reduce inflammation and anti-brain antibodies to protect the brain tissue.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 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.
文摘Background Although traumatic brain injury can lead to opening the blood-brain barrier and leaking of blood substances （including water） into brain tissue, few studies of brain antigens leaking into the blood and the pathways have been reported. Brain antigens result in damage to brain tissues by stimulating the immune system to produce anti-brain antibodies, but no treatment has been reported to reduce the production of anti-brain antibodies and protect the brain tissue. The aim of the study is to confirm the relationship between immune injury and arachnoid granulations following traumatic brain injury, and provide some new methods to inhibit the immune injury.Methods In part one, methylene blue was injected into the rabbits’ cisterna magna after traumatic brain injury, and concentrations of methylene blue and tumor necrosis factor （TNF）-α in blood were detected to determine the permeability of arachnoid granulations. In part two, umbilical cord mesenchymal stem cells and immature dendritic cells were injected into veins, and concentrations of interleukin 1 （IL-1）, IL-10, interferon （IFN）-γ, transforming growth factor （TGF）-β, anti-brain antibodies （ABAb）, and IL-12 were measured by ELISA on days 1, 3, 7, 14 and 21 after injury, and the numbers of leukocytes in the blood were counted. Twenty-one days after injury, expression of glutamate in brain tissue was determined by immunohistochemical staining, and neuronal degeneration was detected by H＆E staining. Results In part one, blood concentrations of methylene blue and TNF-α in the traumatic brain injury group were higher than in the control group （P 〈0.05）. Concentrations of methylene blue and TNF-α in the trauma cerebrospinal fluid （CSF） injected group were higher than in the control cerebrospinal fluid injected group （P 〈0.05）. In part two, concentrations of IL-1, IFN-γ, ABAb, IL-12, expression of glutamate （Glu）, neuronal degeneration and number of peripheral blood leukocytes were lower in the group with cell treatment compared to the control group. IL-10 and TGF-β were elevated compared to the control group. Conclusions Traumatic brain injury can lead to stronger arachnoid granulations （AGs） permeability; umbilical cord mesenchymal stem cells and immature dendritic cells can induce immune tolerance and reduce inflammation and anti-brain antibodies to protect the brain tissue.
