Background:Accurate quantification of 17-hydroxyprogesterone(17-OHP)in serum is vital for clinical and research applications.However,inter-laboratory variability in test results exists owing to the lack of a standardi...Background:Accurate quantification of 17-hydroxyprogesterone(17-OHP)in serum is vital for clinical and research applications.However,inter-laboratory variability in test results exists owing to the lack of a standardized reference measurement procedure(RMP).Therefore,in this study,we developed a highly accurate,cost-effective,and user-friendly candidate RMP(cRMP)for analyzing 17-OHP in serum.Methods:We quantified 17-OHP in serum using a one-step liquid–liquid extraction method with the addition of 17-OHP-^(13)C_(3),followed by liquid chromatography–tandem mass spectrometry.The ability of these methods to suppress interference was evaluated by chromatographic analysis.We assessed accuracy,specificity,the lower limit of quantitation,linearity,and matrix effects by following the standards specified by the Clinical and Laboratory Standards Institute.The consistency between the developed cRMP and the chemiluminescence method was evaluated through experiments with 120 clinical samples.Results:The developed cRMP required only 8 min for accurate quantification of serum 17-OHP without bias from matrix effects or interference from 19 metabolites added as potential interferents.The method exhibited favorable measurement performance,with a quantitation limit of 0.086 ng/mL,linear range of 0.1–400 ng/mL,a total imprecision of≤2.90%,spike recovery of 100.1%–100.6%,and relative deviations from assigned target values(RfB Institution)of−2.91%to 1.10%.The cRMP demonstrated good consistency with the conventional assay(chemiluminescence method),with a correlation coefficient R of 0.96977.Conclusion:A cRMP with high accuracy,cost-effectiveness,and convenient operation was developed for quantifying 17-OHP in serum.Its simplicity and robust performance make it an invaluable addition to the arsenal of analytical tools available for laboratories.This method can enhance the accuracy and reliability of 17-OHP measurements across various laboratories.展开更多
卵巢衰老分为生理性与病理性两类,表现为卵巢储备功能下降、卵泡数量减少和激素水平紊乱,并与多种慢性疾病风险相关。研究发现,免疫失衡在卵巢衰老中扮演核心角色,尤其是辅助性T细胞17(helper T cell 17,Th17细胞)与调节性T细胞(regulat...卵巢衰老分为生理性与病理性两类,表现为卵巢储备功能下降、卵泡数量减少和激素水平紊乱,并与多种慢性疾病风险相关。研究发现,免疫失衡在卵巢衰老中扮演核心角色,尤其是辅助性T细胞17(helper T cell 17,Th17细胞)与调节性T细胞(regulatory T cell,Treg细胞)的比例失调。Th17/Treg平衡受炎症因子、核因子κB(nuclear factor-κB,NF-κB)通路及哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路的精密调控,其失衡会加剧卵巢局部炎症,加速卵泡耗竭。目前,针对Th17/Treg比例失衡的干预策略在自身免疫性疾病的治疗中已取得进展,为卵巢衰老的临床干预提供了新思路。综述Th17/Treg比例失衡在卵巢衰老中的核心作用,为探索延缓卵巢衰老的免疫治疗策略提供了理论依据。展开更多
基金Science and Technology Project of Dongguan,Grant/Award Number:20211800905072Graduate Education Innovation Program Project of Guangdong Province,Grant/Award Number:2022KCXTD010+6 种基金Discipline Construction Project of Guangdong Medical University,Grant/Award Numbers:4SG22098G,4SG22259G,4SG23030G,4SG23143GNational Natural Science Foundation of China,Grant/Award Numbers:81870016,82270013Guangdong Basic and Applied Basic Research Foundation,Grant/Award Number:2022A1515010525Guangdong Medical UniversityShenzhen YHLO Biotech Co.,Ltd.Huazhong University of Science and TechnologyShenzhen Maternal and Child Health Care Hospital。
文摘Background:Accurate quantification of 17-hydroxyprogesterone(17-OHP)in serum is vital for clinical and research applications.However,inter-laboratory variability in test results exists owing to the lack of a standardized reference measurement procedure(RMP).Therefore,in this study,we developed a highly accurate,cost-effective,and user-friendly candidate RMP(cRMP)for analyzing 17-OHP in serum.Methods:We quantified 17-OHP in serum using a one-step liquid–liquid extraction method with the addition of 17-OHP-^(13)C_(3),followed by liquid chromatography–tandem mass spectrometry.The ability of these methods to suppress interference was evaluated by chromatographic analysis.We assessed accuracy,specificity,the lower limit of quantitation,linearity,and matrix effects by following the standards specified by the Clinical and Laboratory Standards Institute.The consistency between the developed cRMP and the chemiluminescence method was evaluated through experiments with 120 clinical samples.Results:The developed cRMP required only 8 min for accurate quantification of serum 17-OHP without bias from matrix effects or interference from 19 metabolites added as potential interferents.The method exhibited favorable measurement performance,with a quantitation limit of 0.086 ng/mL,linear range of 0.1–400 ng/mL,a total imprecision of≤2.90%,spike recovery of 100.1%–100.6%,and relative deviations from assigned target values(RfB Institution)of−2.91%to 1.10%.The cRMP demonstrated good consistency with the conventional assay(chemiluminescence method),with a correlation coefficient R of 0.96977.Conclusion:A cRMP with high accuracy,cost-effectiveness,and convenient operation was developed for quantifying 17-OHP in serum.Its simplicity and robust performance make it an invaluable addition to the arsenal of analytical tools available for laboratories.This method can enhance the accuracy and reliability of 17-OHP measurements across various laboratories.
文摘卵巢衰老分为生理性与病理性两类,表现为卵巢储备功能下降、卵泡数量减少和激素水平紊乱,并与多种慢性疾病风险相关。研究发现,免疫失衡在卵巢衰老中扮演核心角色,尤其是辅助性T细胞17(helper T cell 17,Th17细胞)与调节性T细胞(regulatory T cell,Treg细胞)的比例失调。Th17/Treg平衡受炎症因子、核因子κB(nuclear factor-κB,NF-κB)通路及哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)通路的精密调控,其失衡会加剧卵巢局部炎症,加速卵泡耗竭。目前,针对Th17/Treg比例失衡的干预策略在自身免疫性疾病的治疗中已取得进展,为卵巢衰老的临床干预提供了新思路。综述Th17/Treg比例失衡在卵巢衰老中的核心作用,为探索延缓卵巢衰老的免疫治疗策略提供了理论依据。
