Inosine is a vital RNA modification across three kingdoms of life.It has been demonstrated that inosine plays important roles in modulation of the fate of RNAs.In the current study,we developed a highly sensitive meth...Inosine is a vital RNA modification across three kingdoms of life.It has been demonstrated that inosine plays important roles in modulation of the fate of RNAs.In the current study,we developed a highly sensitive method to determine inosine in a single cell by N-cyclohexyl-N’-β-(4-methylmorpholinium)ethylcarbodiimide p-toluenesulfonate(CMCT)derivatization in combination with mass spectrometry analysis.The results showed that the detection sensitivity of inosine was increased by 556-fold after CMCT derivatization,with the limit of detection(LOD)being 4.5 amol.With the established method,we could detect inosine from 13.0 pg of total RNA of HEK293T cells.Meanwhile,inosine in RNA from a single cell could also be clearly detected due to the improved detection sensitivity.Moreover,we found the level of inosine in RNA of sleep-deprived mice was significantly increased compared to the control mice,indicating that inosine is associated with sleep behavior and might be a potential indicator of sleep disorder.Taken together,the chemical derivatization coupled with mass spectrometry analysis offers a valuable tool in determination of endogenous RNA modifications in a single cell,which should benefit the functional study of RNA modification in rare clinical samples.展开更多
文摘目的 探讨中医针灸辅助现代康复治疗脑梗死患者的疗效观察。方法 选取2018年1月—2022年2月期间张家港市中医医院收治的100例脑梗死患者,按随机数字表法分为对照组与试验组,每组各50例。两组患者均给予脑梗死基础治疗,病情稳定后开展康复训练,在此基础上,对照组给予低频重复经颅磁刺激,试验组给予中医针灸辅助重复经颅磁刺激,均治疗4周。观察比较两组患者临床疗效、中医证候疗效及治疗前后肢体运动功能(Fugl-Meyer Assessment, FMA)、运动诱发电位(Motor evoked potential, MEP)波幅、潜伏期、中枢运动传导时间(Central motor conduction time, CMCT)、神经功能评分[美国国立卫生院卒中量表(National Institutes of Health Stroke Scale, NIHSS)]、自理能力(Self-care ability in daily life, ADL)变化。结果 治疗后试验组临床疗效总有效率95.92%(47/49)优于对照组83.33%(40/48),差异有统计学意义(P<0.05)。治疗后试验组中医证候疗效总有效率91.84%(45/49)优于对照组72.92%(35/48),差异有统计学意义(P<0.05)。治疗后两组患者FMA上、下肢评分及总分均较治疗前升高,差异有统计学意义(P<0.05),且试验组FMA上、下肢评分及总分明显高于对照组,差异有统计学意义(P<0.05)。治疗后两组患者MEP波幅、潜伏期及CMCT均较治疗前改善,差异有统计学意义(P<0.05),且试验组优于对照组,差异有统计学意义(P<0.05)。治疗后两组患者NIHSS评分及ADL评分均较治疗前降低,差异有统计学意义(P<0.05),且试验组低于对照组,差异有统计学意义(P<0.05)。结论 中医针灸辅助现代康复技术治疗脑梗死效果显著,有利于改善患者肢体运动功能、MEP、CMCT、神经功能及生活自理能力。
基金supported by the National Key R&D Program of China(Nos.2022YFA0806600,2022YFC3400700)the National Natural Science Foundation of China(Nos.22277093,22074110,21721005).
文摘Inosine is a vital RNA modification across three kingdoms of life.It has been demonstrated that inosine plays important roles in modulation of the fate of RNAs.In the current study,we developed a highly sensitive method to determine inosine in a single cell by N-cyclohexyl-N’-β-(4-methylmorpholinium)ethylcarbodiimide p-toluenesulfonate(CMCT)derivatization in combination with mass spectrometry analysis.The results showed that the detection sensitivity of inosine was increased by 556-fold after CMCT derivatization,with the limit of detection(LOD)being 4.5 amol.With the established method,we could detect inosine from 13.0 pg of total RNA of HEK293T cells.Meanwhile,inosine in RNA from a single cell could also be clearly detected due to the improved detection sensitivity.Moreover,we found the level of inosine in RNA of sleep-deprived mice was significantly increased compared to the control mice,indicating that inosine is associated with sleep behavior and might be a potential indicator of sleep disorder.Taken together,the chemical derivatization coupled with mass spectrometry analysis offers a valuable tool in determination of endogenous RNA modifications in a single cell,which should benefit the functional study of RNA modification in rare clinical samples.
