BACKGROUND Febrile seizures(FS)are common pediatric neurological disorders typically triggered by fever,affecting children aged 6 months to 5 years.Although usually benign,early and accurate diagnosis is crucial for e...BACKGROUND Febrile seizures(FS)are common pediatric neurological disorders typically triggered by fever,affecting children aged 6 months to 5 years.Although usually benign,early and accurate diagnosis is crucial for effective management.AIM To investigate the diagnostic potential of interleukin-1 beta(IL-1β)and micro-RNA-146a(miR-146a)in distinguishing FS from healthy subjects(HS),fever-only(FO),and seizure-only(SO)cases.METHODS Pediatric patients were grouped into FS(n=68),HS(n=52),FO(n=52),and SO(n=32).Blood samples were collected within 30 minutes of seizure onset or fever presentation.Plasma samples were analyzed for IL-1β(enzyme-linked immuno-sorbent assay)and miR-146a(real-time polymerase chain reaction).Group comparisons were conducted using analysis of variance or the non-parametric alternative(Kruskal-Wallis test),while diagnostic performance was evaluated through receiver operating characteristic curve analysis.RESULTS IL-1βlevels showed the highest level in FS,followed by FO and SO,with HS showing the lowest levels.IL-1βdistinguished FS from HS with an area under the curve(AUC)of 0.9983(sensitivity:100%,specificity:96.15%).miR-146a was lowest in FS and highest in HS,achieving an AUC of 1.000(sensitivity:100%,specificity:98.08%).IL-1βand miR-146a also effectively differentiated FS from FO and SO,with respective AUCs of 0.8862 and 0.8173 for FS vs FO.CONCLUSION IL-1βand miR-146a could be promising diagnostic biomarkers for FS,distinguishing FS from HS,FO,and SO with high sensitivity and specificity.These markers possess the potential to enhance early FS diagnosis.展开更多
Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis...Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis,and restenosis.MicroRNA-146a(miR-146a)has been proven to be involved in cell proliferation,migration,and tumor metabolism.However,little is known about the functional role of miR-146a in VSMC differentiation from embryonic stem cells(ESCs).This study aimed to determine the role of miR-146a in VSMC differentiation from ESCs.Methods Mouse ESCs were differentiated into VSMCs,and the cell extracts were analyzed by Western blotting and RT-qPCR.In addition,luciferase reporter assays using ESCs transfected with miR-146a/mimic and plasmids were performed.Finally,C57BL/6J female mice were injected with mimic or miR-146a-overexpressing ESCs,and immunohistochemistry,Western blotting,and RT-qPCR assays were carried out on tissue samples from these mice.Results miR-146a was significantly upregulated during VSMC differentiation,accompanied with the VSMC-specific marker genes smooth muscle-alpha-actin(SMαA),smooth muscle 22(SM22),smooth muscle myosin heavy chain(SMMHC),and h1-calponin.Furthermore,overexpression of miR-146a enhanced the differentiation process in vitro and in vivo.Concurrently,the expression of Kruppel-like factor 4(KLF4),predicted as one of the top targets of miR-146a,was sharply decreased in miR-146a-overexpressing ESCs.Importantly,inhibiting KLF4 expression enhanced the VSMC-specific gene expression induced by miR-146a overexpression in differentiating ESCs.In addition,miR-146a upregulated the mRNA expression levels and transcriptional activity of VSMC differentiation-related transcription factors,including serum response factor(SRF)and myocyte enhancer factor 2c(MEF-2c).Conclusion Our data support that miR-146a promotes ESC-VSMC differentiation through regulating KLF4 and modulating the transcription factor activity of VSMCs.展开更多
基金Supported by the Department of Research and Technology,Golestan University of Medical Sciences,Iran,No.113610the Oman Ministry of Higher Education,Research,and Innovation,No.BFP/RGP/HSS/24/015.
摘要BACKGROUND Febrile seizures(FS)are common pediatric neurological disorders typically triggered by fever,affecting children aged 6 months to 5 years.Although usually benign,early and accurate diagnosis is crucial for effective management.AIM To investigate the diagnostic potential of interleukin-1 beta(IL-1β)and micro-RNA-146a(miR-146a)in distinguishing FS from healthy subjects(HS),fever-only(FO),and seizure-only(SO)cases.METHODS Pediatric patients were grouped into FS(n=68),HS(n=52),FO(n=52),and SO(n=32).Blood samples were collected within 30 minutes of seizure onset or fever presentation.Plasma samples were analyzed for IL-1β(enzyme-linked immuno-sorbent assay)and miR-146a(real-time polymerase chain reaction).Group comparisons were conducted using analysis of variance or the non-parametric alternative(Kruskal-Wallis test),while diagnostic performance was evaluated through receiver operating characteristic curve analysis.RESULTS IL-1βlevels showed the highest level in FS,followed by FO and SO,with HS showing the lowest levels.IL-1βdistinguished FS from HS with an area under the curve(AUC)of 0.9983(sensitivity:100%,specificity:96.15%).miR-146a was lowest in FS and highest in HS,achieving an AUC of 1.000(sensitivity:100%,specificity:98.08%).IL-1βand miR-146a also effectively differentiated FS from FO and SO,with respective AUCs of 0.8862 and 0.8173 for FS vs FO.CONCLUSION IL-1βand miR-146a could be promising diagnostic biomarkers for FS,distinguishing FS from HS,FO,and SO with high sensitivity and specificity.These markers possess the potential to enhance early FS diagnosis.
基金funded by the National Natural Science Foundation of China(No.82070376 and No.81873491)the Natural Science Foundation of Zhejiang Province(No.LY21H020005)+1 种基金the Zhejiang Medical Science and Technology Project(No.2019KY376 and No.2018KY071)a Ningbo Science and Technology Project(No.202002N3173).
摘要Objective Vascular smooth muscle cell(VSMC)differentiation from stem cells is one source of the increasing number of VSMCs that are involved in vascular remodeling-related diseases such as hypertension,atherosclerosis,and restenosis.MicroRNA-146a(miR-146a)has been proven to be involved in cell proliferation,migration,and tumor metabolism.However,little is known about the functional role of miR-146a in VSMC differentiation from embryonic stem cells(ESCs).This study aimed to determine the role of miR-146a in VSMC differentiation from ESCs.Methods Mouse ESCs were differentiated into VSMCs,and the cell extracts were analyzed by Western blotting and RT-qPCR.In addition,luciferase reporter assays using ESCs transfected with miR-146a/mimic and plasmids were performed.Finally,C57BL/6J female mice were injected with mimic or miR-146a-overexpressing ESCs,and immunohistochemistry,Western blotting,and RT-qPCR assays were carried out on tissue samples from these mice.Results miR-146a was significantly upregulated during VSMC differentiation,accompanied with the VSMC-specific marker genes smooth muscle-alpha-actin(SMαA),smooth muscle 22(SM22),smooth muscle myosin heavy chain(SMMHC),and h1-calponin.Furthermore,overexpression of miR-146a enhanced the differentiation process in vitro and in vivo.Concurrently,the expression of Kruppel-like factor 4(KLF4),predicted as one of the top targets of miR-146a,was sharply decreased in miR-146a-overexpressing ESCs.Importantly,inhibiting KLF4 expression enhanced the VSMC-specific gene expression induced by miR-146a overexpression in differentiating ESCs.In addition,miR-146a upregulated the mRNA expression levels and transcriptional activity of VSMC differentiation-related transcription factors,including serum response factor(SRF)and myocyte enhancer factor 2c(MEF-2c).Conclusion Our data support that miR-146a promotes ESC-VSMC differentiation through regulating KLF4 and modulating the transcription factor activity of VSMCs.