Background:Multiple mitochondrial dysfunction syndromes(MMDS)are rare mitochondrial diseases caused by mutation of mitochondrial iron–sulfur cluster synthesis proteins.This study established a rat model simulating MM...Background:Multiple mitochondrial dysfunction syndromes(MMDS)are rare mitochondrial diseases caused by mutation of mitochondrial iron–sulfur cluster synthesis proteins.This study established a rat model simulating MMDS5 disease in the nervous system to investigate its pathological features and neuronal death.Methods:We generated neuron-specific Isca1 knockout rat(Isca1 flox/flox-NeuN-Cre)using CRISPR-Cas9 technology.The brain structure changes of CKO rats were studied with MRI,and the behavior abnormalities were analyzed through gait analysis and open field tests,Y maze tests and food maze tests.The pathological changes of neurons were analyzed through H&E staining,Nissl staining,and Golgi staining.Mitochondrial damage was assessed by TEM,western blot and ATP assay,and the morphology of neurons was assessed by WGA immunofluorescence to detect the death of neurons.Results:This study established the disease model of MMDS5 in the nervous system for the first time,and found that after Isca1 loss,the rats suffered from developmental retardation,epilepsy,memory impairment,massive neuronal death,reduced number of Nissl bodies and dendritic spines,mitochondrial fragmentation,cristae fracture,reduced content of respiratory chain complex protein,and reduced production of ATP.Isca1 knockout caused neuronal oncosis.Conclusions:This rat model can be used to study the pathogenesis of MMDS.In addition,compared with human MMDS5,the rat model can survive up to 8 weeks of age,effectively extending the window of clinical treatment research,and can be used for the treatment of neurological symptoms in other mitochondrial diseases.展开更多
Background : Iron-sulfur cluster assembly 1( ISCA 1) is an iron-sulfur(Fe/S) carrier protein that accepts Fe/S from a scaffold protein and transfers it to target proteins including the mitochondrial Fe/S containing pr...Background : Iron-sulfur cluster assembly 1( ISCA 1) is an iron-sulfur(Fe/S) carrier protein that accepts Fe/S from a scaffold protein and transfers it to target proteins including the mitochondrial Fe/S containing proteins. ISCA 1 is also the newly identified causal gene for multiple mitochondrial dysfunctions syndrome( MMDS). However, our knowledge about the physiological function of ISCA 1 in vivo is currently limited. In this study, we generated an ISCA 1 knockout rat line and analyzed the embryo development. Methods : ISCA 1 knockout rats were generated by replacing the exon1 of ISCA 1 gene with the mC herry-Cre fusion gene using CRISPR-Cas9 technology. The ISCA 1 expression pattern was analyzed by fluorescence imaging using ISCA 1 promotor driven Cre and mC herry expression. The embryonic morphology was examinated by microscope and mitochondrial proteins were tested by Western blot. Results : An ISCA 1 knockout rat line was obtained, which expressed mC herry-Cre fusion protein. Both of the fluorescence images from mC herry and Cre induced mC herry in a reporter rat strain, showing that ISCA 1 expressed in most of the tissues in rats. The ISCA 1 knockout resulted in abnormal development at 8.5 days, with a significant decrease of NDUFA 9 protein and an increase of aconitase 2( ACO 2) in rat embryos. Conclusion : Deletion of ISCA 1 induced early death in rats. ISCA 1 affected the expression of key proteins in the mitochondrial respiratory chain complex, suggesting that ISCA 1 has an important influence on the respiratory complex and energy metabolism.展开更多
本文总结和审视现代信息技术应用于会计工作的内涵和作用,根据目前我国信息化现状及未来可能的实践提出建立和实施会计管理信息化的ISCA(Information System,Control and Auditing)模型:首先是建立在企业管理信息系统环境中的会计信息系...本文总结和审视现代信息技术应用于会计工作的内涵和作用,根据目前我国信息化现状及未来可能的实践提出建立和实施会计管理信息化的ISCA(Information System,Control and Auditing)模型:首先是建立在企业管理信息系统环境中的会计信息系统,是一种事件驱动模式的系统,其核心是集成;同时,为了确保会计信息系统(AIS)安全有效地运作,必须建立健全的信息系统内部控制制度;并且,为了确保和审查内部控制制度的有效执行,必须开展对AIS及其内控制度的审计,以最终地达到对AIS安全、可靠、有效和高效地应用.三者的有机结合构成了AIS的ISCA模型。正确地实施ISCA模型,可实现企业的物流、业务流、资金流、信息流、控制过程和审计过程的整合和集成,使现代信息技术应用于会计管理工作并取得较好的效果。展开更多
基金CAMS Innovation Fund for Medical Sciences CIFMSGrant/Award Number:2021-I2M-1-034+3 种基金National Key Research and Development Program of ChinaGrant/Award Number:2022YFF0710702National Natural Science Foundation of ChinaGrant/Award Number:31970508。
文摘Background:Multiple mitochondrial dysfunction syndromes(MMDS)are rare mitochondrial diseases caused by mutation of mitochondrial iron–sulfur cluster synthesis proteins.This study established a rat model simulating MMDS5 disease in the nervous system to investigate its pathological features and neuronal death.Methods:We generated neuron-specific Isca1 knockout rat(Isca1 flox/flox-NeuN-Cre)using CRISPR-Cas9 technology.The brain structure changes of CKO rats were studied with MRI,and the behavior abnormalities were analyzed through gait analysis and open field tests,Y maze tests and food maze tests.The pathological changes of neurons were analyzed through H&E staining,Nissl staining,and Golgi staining.Mitochondrial damage was assessed by TEM,western blot and ATP assay,and the morphology of neurons was assessed by WGA immunofluorescence to detect the death of neurons.Results:This study established the disease model of MMDS5 in the nervous system for the first time,and found that after Isca1 loss,the rats suffered from developmental retardation,epilepsy,memory impairment,massive neuronal death,reduced number of Nissl bodies and dendritic spines,mitochondrial fragmentation,cristae fracture,reduced content of respiratory chain complex protein,and reduced production of ATP.Isca1 knockout caused neuronal oncosis.Conclusions:This rat model can be used to study the pathogenesis of MMDS.In addition,compared with human MMDS5,the rat model can survive up to 8 weeks of age,effectively extending the window of clinical treatment research,and can be used for the treatment of neurological symptoms in other mitochondrial diseases.
基金CAMS Innovation Fund for Medical Sciences,Grant/Award Number:2017-I2M-3-015 and 2016-I2M-1-004
文摘Background : Iron-sulfur cluster assembly 1( ISCA 1) is an iron-sulfur(Fe/S) carrier protein that accepts Fe/S from a scaffold protein and transfers it to target proteins including the mitochondrial Fe/S containing proteins. ISCA 1 is also the newly identified causal gene for multiple mitochondrial dysfunctions syndrome( MMDS). However, our knowledge about the physiological function of ISCA 1 in vivo is currently limited. In this study, we generated an ISCA 1 knockout rat line and analyzed the embryo development. Methods : ISCA 1 knockout rats were generated by replacing the exon1 of ISCA 1 gene with the mC herry-Cre fusion gene using CRISPR-Cas9 technology. The ISCA 1 expression pattern was analyzed by fluorescence imaging using ISCA 1 promotor driven Cre and mC herry expression. The embryonic morphology was examinated by microscope and mitochondrial proteins were tested by Western blot. Results : An ISCA 1 knockout rat line was obtained, which expressed mC herry-Cre fusion protein. Both of the fluorescence images from mC herry and Cre induced mC herry in a reporter rat strain, showing that ISCA 1 expressed in most of the tissues in rats. The ISCA 1 knockout resulted in abnormal development at 8.5 days, with a significant decrease of NDUFA 9 protein and an increase of aconitase 2( ACO 2) in rat embryos. Conclusion : Deletion of ISCA 1 induced early death in rats. ISCA 1 affected the expression of key proteins in the mitochondrial respiratory chain complex, suggesting that ISCA 1 has an important influence on the respiratory complex and energy metabolism.
文摘本文总结和审视现代信息技术应用于会计工作的内涵和作用,根据目前我国信息化现状及未来可能的实践提出建立和实施会计管理信息化的ISCA(Information System,Control and Auditing)模型:首先是建立在企业管理信息系统环境中的会计信息系统,是一种事件驱动模式的系统,其核心是集成;同时,为了确保会计信息系统(AIS)安全有效地运作,必须建立健全的信息系统内部控制制度;并且,为了确保和审查内部控制制度的有效执行,必须开展对AIS及其内控制度的审计,以最终地达到对AIS安全、可靠、有效和高效地应用.三者的有机结合构成了AIS的ISCA模型。正确地实施ISCA模型,可实现企业的物流、业务流、资金流、信息流、控制过程和审计过程的整合和集成,使现代信息技术应用于会计管理工作并取得较好的效果。
