In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection a...In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection and analysis of one or two genes cannot fully reveal the complex molecular biological mechanisms of spinal cord injury.The emergence of second-generation sequencing technology(RNA sequencing)has provided a better method.In this study,RNA sequencing technology was used to analyze differentially expressed genes at different time points after spinal cord injury in rat models established by contusion of the eighth thoracic segment.The numbers of genes that changed significantly were 944,1362 and 1421 at 1,4 and 7 days after spinal cord injury respectively.After gene ontology analysis and temporal expression analysis of the differentially expressed genes,C5ar1,Socs3 and CCL6 genes were then selected and identified by real-time polymerase chain reaction and western blot assay.The mRNA expression trends of C5ar1,Socs3 and CCL6 genes were consistent with the RNA sequencing results.Further verification and analysis of C5ar1 indicate that the level of protein expression of C5ar1 was consistent with its nucleic acid level after spinal cord injury.C5ar1 was mainly expressed in neurons and astrocytes.Finally,the gene Itgb2,which may be related to C5ar1,was found by Chilibot database and literature search.Immunofluorescence histochemical results showed that the expression of Itgb2 was highly consistent with that of C5ar1.Itgb2 was expressed in astrocytes.RNA sequencing technology can screen differentially expressed genes at different time points after spinal cord injury.Through analysis and verification,genes strongly associated with spinal cord injury can be screened.This can provide experimental data for further determining the molecular mechanism of spinal cord injury,and also provide possible targets for the treatment of spinal cord injury.This study was approved ethically by the Laboratory Animal Ethics Committee of Jiangsu Province,China(approval No.2018-0306-001)on March 6,2018.展开更多
Objective:To investigate the role of T helper 9(Th9) cells in liver cirrhosis(LC) patients and whether chemokine receptor type 6(CCR6)/chemokine ligand 20(CCL20) axis is involving in the recruitment of Th9 cells into ...Objective:To investigate the role of T helper 9(Th9) cells in liver cirrhosis(LC) patients and whether chemokine receptor type 6(CCR6)/chemokine ligand 20(CCL20) axis is involving in the recruitment of Th9 cells into liver.Methods:Peripheral blood and liver tissue from 30 LC patients and 18 normal controls were recruited.The frequency of Th9 cells and CCR4,CCR6 in the peripheral blood was tested by flow cytometry.Serum interleukin(IL)-9 and CCL20 levels were tested by enzyme-linked immunosorbent assay.Immunohistochemical staining was used to detect a-smooth muscle actin,CCR6 and CCL20 expression in liver tissue.Results:The frequency of Th9 cells in LC patients was significantly increased compared with controls(P < 0.05).The serum IL-9 level and CCL20 level increased markedly in LC patients compared with controls(P < 0.05),and IL-9 was positively correlated to Th9 cells and CCL20.Furthermore,the frequency of Th9 cells was correlated to prothrombin time,total bilirubin level,hyaluronic acid and type IV collagen in LC patients.We also found that Th9 cells in LC patients expressed higher frequency of CCR4+,CCR6+(P < 0.05).Compared with normal controls,the expression of CCR6 and CCL20 in LC tissue were significantly elevated(P < 0.05).The expression of a-smooth muscle actin was correlated to the CCR6 and CCL20 in liver tissue of LC patients.Conclusions:This study suggests that Th9 cells may participate in the pathogenesis of LC,and the recruitment of Th9 cells into liver tissue might be through CCL20/CCR6 axis.展开更多
基金supported by the National Natural Science Foundation of China,No.31570983(to XDW)the Priority Academic Program Development of Jiangsu Higher Education Institutes of China
文摘In the search for a therapeutic schedule for spinal cord injury,it is necessary to understand key genes and their corresponding regulatory networks involved in the spinal cord injury process.However,ad hoc selection and analysis of one or two genes cannot fully reveal the complex molecular biological mechanisms of spinal cord injury.The emergence of second-generation sequencing technology(RNA sequencing)has provided a better method.In this study,RNA sequencing technology was used to analyze differentially expressed genes at different time points after spinal cord injury in rat models established by contusion of the eighth thoracic segment.The numbers of genes that changed significantly were 944,1362 and 1421 at 1,4 and 7 days after spinal cord injury respectively.After gene ontology analysis and temporal expression analysis of the differentially expressed genes,C5ar1,Socs3 and CCL6 genes were then selected and identified by real-time polymerase chain reaction and western blot assay.The mRNA expression trends of C5ar1,Socs3 and CCL6 genes were consistent with the RNA sequencing results.Further verification and analysis of C5ar1 indicate that the level of protein expression of C5ar1 was consistent with its nucleic acid level after spinal cord injury.C5ar1 was mainly expressed in neurons and astrocytes.Finally,the gene Itgb2,which may be related to C5ar1,was found by Chilibot database and literature search.Immunofluorescence histochemical results showed that the expression of Itgb2 was highly consistent with that of C5ar1.Itgb2 was expressed in astrocytes.RNA sequencing technology can screen differentially expressed genes at different time points after spinal cord injury.Through analysis and verification,genes strongly associated with spinal cord injury can be screened.This can provide experimental data for further determining the molecular mechanism of spinal cord injury,and also provide possible targets for the treatment of spinal cord injury.This study was approved ethically by the Laboratory Animal Ethics Committee of Jiangsu Province,China(approval No.2018-0306-001)on March 6,2018.
基金Supported by grants from the National Natural Science Foundation of China(81260083,31360221)Natural Science Foundation of Guangxi Province(2014GXNSFAA118203)
文摘Objective:To investigate the role of T helper 9(Th9) cells in liver cirrhosis(LC) patients and whether chemokine receptor type 6(CCR6)/chemokine ligand 20(CCL20) axis is involving in the recruitment of Th9 cells into liver.Methods:Peripheral blood and liver tissue from 30 LC patients and 18 normal controls were recruited.The frequency of Th9 cells and CCR4,CCR6 in the peripheral blood was tested by flow cytometry.Serum interleukin(IL)-9 and CCL20 levels were tested by enzyme-linked immunosorbent assay.Immunohistochemical staining was used to detect a-smooth muscle actin,CCR6 and CCL20 expression in liver tissue.Results:The frequency of Th9 cells in LC patients was significantly increased compared with controls(P < 0.05).The serum IL-9 level and CCL20 level increased markedly in LC patients compared with controls(P < 0.05),and IL-9 was positively correlated to Th9 cells and CCL20.Furthermore,the frequency of Th9 cells was correlated to prothrombin time,total bilirubin level,hyaluronic acid and type IV collagen in LC patients.We also found that Th9 cells in LC patients expressed higher frequency of CCR4+,CCR6+(P < 0.05).Compared with normal controls,the expression of CCR6 and CCL20 in LC tissue were significantly elevated(P < 0.05).The expression of a-smooth muscle actin was correlated to the CCR6 and CCL20 in liver tissue of LC patients.Conclusions:This study suggests that Th9 cells may participate in the pathogenesis of LC,and the recruitment of Th9 cells into liver tissue might be through CCL20/CCR6 axis.
