Nitric oxide(NO)participates in various pathways and revealing its dynamics is critical for resolving its pathophysiology.While there are methods available for detecting biological NO,few are capable of tracking NO dy...Nitric oxide(NO)participates in various pathways and revealing its dynamics is critical for resolving its pathophysiology.While there are methods available for detecting biological NO,few are capable of tracking NO dynamics.Herein,inspired by the cellular machinery of reversible thiol modification by NO,we have successfully designed a family of cysteine analogues tagged with fluorophores for visualizing cellular NO dynamics.展开更多
目的研究腺病毒携带目的基因经小鼠耳后入路圆窗膜显微注射途径耳蜗转导的可行性,为以小鼠作为动物模型的内耳基因治疗提供实验基础和解剖学依据。方法12只C57BL/6J小鼠分为2组,实验组(8只)以重组腺病毒携带的增强型绿色荧光蛋白基因(en...目的研究腺病毒携带目的基因经小鼠耳后入路圆窗膜显微注射途径耳蜗转导的可行性,为以小鼠作为动物模型的内耳基因治疗提供实验基础和解剖学依据。方法12只C57BL/6J小鼠分为2组,实验组(8只)以重组腺病毒携带的增强型绿色荧光蛋白基因(enhanced green fluorescent protein,EGFP)、对照组(4只)以人工外淋巴液经耳后入路圆窗膜显微注射注入耳蜗内。分别于术后5、14天取双侧耳蜗标本做基底膜铺片,在激光共聚焦显微镜下观察GFP表达。结果术后动物存活10只(每组死亡1只)。实验组转染后耳蜗底回基底膜及螺旋神经节上目的基因有表达,14天组强于5天组。对照组耳蜗未见荧光表达。结论耳后入路操作简单、损伤小、易于暴露圆窗龛。耳后入路圆窗膜显微注射腺病毒携带目的基因转导的方法能够将目的基因成功转导至耳蜗组织并表达。展开更多
基金supported by the National Natural Science Foundations of China(nos.22077112,21778048,81673489,31871414,and 81125023)Natural Science Foundation of Zhejiang Province,China(no.LR18H300001)National Science and Technology Major Project“Key New Drug Creation and Manufacturing Program”(nos.2018ZX09711002-010-004,2018ZX09711002-007-002,2019ZX09201001-003-009,2019ZX09201001-003-010,and 2019ZX09201001-004-010),K.C.Wong Education Foundation,and Singapore University of Technology and Design(SUTD)[SUTD-ZJU IDEA grant nos.T1SRCI17126 and SUTD-ZJU(VP)201905].
文摘Nitric oxide(NO)participates in various pathways and revealing its dynamics is critical for resolving its pathophysiology.While there are methods available for detecting biological NO,few are capable of tracking NO dynamics.Herein,inspired by the cellular machinery of reversible thiol modification by NO,we have successfully designed a family of cysteine analogues tagged with fluorophores for visualizing cellular NO dynamics.
文摘目的研究腺病毒携带目的基因经小鼠耳后入路圆窗膜显微注射途径耳蜗转导的可行性,为以小鼠作为动物模型的内耳基因治疗提供实验基础和解剖学依据。方法12只C57BL/6J小鼠分为2组,实验组(8只)以重组腺病毒携带的增强型绿色荧光蛋白基因(enhanced green fluorescent protein,EGFP)、对照组(4只)以人工外淋巴液经耳后入路圆窗膜显微注射注入耳蜗内。分别于术后5、14天取双侧耳蜗标本做基底膜铺片,在激光共聚焦显微镜下观察GFP表达。结果术后动物存活10只(每组死亡1只)。实验组转染后耳蜗底回基底膜及螺旋神经节上目的基因有表达,14天组强于5天组。对照组耳蜗未见荧光表达。结论耳后入路操作简单、损伤小、易于暴露圆窗龛。耳后入路圆窗膜显微注射腺病毒携带目的基因转导的方法能够将目的基因成功转导至耳蜗组织并表达。
