Benign multiple sclerosis is a retrospective diagnosis based primarily on a lack of motor symptom progression. Recent findings that suggest patients with benign multiple sclerosis experience non-motor symptoms highlig...Benign multiple sclerosis is a retrospective diagnosis based primarily on a lack of motor symptom progression. Recent findings that suggest patients with benign multiple sclerosis experience non-motor symptoms highlight the need for a more prospective means to diagnose benign multiple sclerosis early in order to help direct patient care. In this study, we present optical coherence tomography and T cell neurotrophin gene analysis findings in a small number of patients with benign multiple sclerosis. Our results demonstrated that retinal nerve fiber layer was mildly thinned, and T cells had a distinct gene expression profile that included upregulation of interleukin 10 and leukemia inhibitory factor, downregulation of interleukin 6 and neurotensin high affinity receptor 1(a novel neurotrophin receptor). These findings add evidence for further investigation into optical coherence tomography and m RNA profiling in larger cohorts as a potential means to diagnose benign multiple sclerosis in a more prospective manner.展开更多
目的构建与海肾荧光素酶（Rluc）融合的人Neurotensin-R1（HumanNeurotensinreceptorl，NTS1 or NTSR1）真核表达载体，用于生物发光共振能量转移法检测人NTS与其它受体间的相互作用以及研究Neurotensin—R介导的细胞内信号转导机制。...目的构建与海肾荧光素酶（Rluc）融合的人Neurotensin-R1（HumanNeurotensinreceptorl，NTS1 or NTSR1）真核表达载体，用于生物发光共振能量转移法检测人NTS与其它受体间的相互作用以及研究Neurotensin—R介导的细胞内信号转导机制。方法以质粒peDNA3．1-hNTS1为模板，PCR方法扩增人NTS。扩增的人NTS以及质粒pRluc—pcDNA3．1用NotI和XbaI双酶切，然后将这2种酶切产物按常规方法连接、转化至大肠杆菌Top10中，该菌在培养箱中孵育12～16h后，挑取菌落培养，提取质粒，进行酶切鉴定，最后进行测序。将测序正确的重组载体用脂质体法转染人胚胎肾（humanembryonickidney293，HEK293）细胞。最后，通过共聚焦显微镜观察经过免疫荧光染色的细胞以及Westernblot鉴定人Neu—rotensinl—R的表达。结果通过PCR扩增出一条1257bp的基因片段，序列与GenBank（NM-002531）相同。Westernblot中大约90kDa处有一蛋白条带，与预期大小相同。人Neurotensin—R表达在细胞膜上。结论成功构建了pRluc—hNeurotensinl—R重组表达载体，建立了该质粒转染HEK293的细胞模型。可被用于检测与其它受体间的相互作用以及研究Neurotensinl—R介导的细胞内信号转导机制，这将有助于探究疾病的发病机制及开发新的药物靶点。展开更多
基金funded by an investigator-initiated,unrestricted research grant(to YMD)from Biogen Idec.YMD served as a consultant and/or received grant support from:Acorda,Bayer Pharmaceutical,EMD Serono,Genzyme,Novartis,Questor,Teva Neuroscience and Chugai Pharmasupported by grants from NIH NIAID Autoimmune Center of Excellence:UM1-AI110557NIH NINDS R01-NS080821,Novartis and Chugai(to YMD)
文摘Benign multiple sclerosis is a retrospective diagnosis based primarily on a lack of motor symptom progression. Recent findings that suggest patients with benign multiple sclerosis experience non-motor symptoms highlight the need for a more prospective means to diagnose benign multiple sclerosis early in order to help direct patient care. In this study, we present optical coherence tomography and T cell neurotrophin gene analysis findings in a small number of patients with benign multiple sclerosis. Our results demonstrated that retinal nerve fiber layer was mildly thinned, and T cells had a distinct gene expression profile that included upregulation of interleukin 10 and leukemia inhibitory factor, downregulation of interleukin 6 and neurotensin high affinity receptor 1(a novel neurotrophin receptor). These findings add evidence for further investigation into optical coherence tomography and m RNA profiling in larger cohorts as a potential means to diagnose benign multiple sclerosis in a more prospective manner.
文摘目的构建与海肾荧光素酶（Rluc）融合的人Neurotensin-R1（HumanNeurotensinreceptorl，NTS1 or NTSR1）真核表达载体，用于生物发光共振能量转移法检测人NTS与其它受体间的相互作用以及研究Neurotensin—R介导的细胞内信号转导机制。方法以质粒peDNA3．1-hNTS1为模板，PCR方法扩增人NTS。扩增的人NTS以及质粒pRluc—pcDNA3．1用NotI和XbaI双酶切，然后将这2种酶切产物按常规方法连接、转化至大肠杆菌Top10中，该菌在培养箱中孵育12～16h后，挑取菌落培养，提取质粒，进行酶切鉴定，最后进行测序。将测序正确的重组载体用脂质体法转染人胚胎肾（humanembryonickidney293，HEK293）细胞。最后，通过共聚焦显微镜观察经过免疫荧光染色的细胞以及Westernblot鉴定人Neu—rotensinl—R的表达。结果通过PCR扩增出一条1257bp的基因片段，序列与GenBank（NM-002531）相同。Westernblot中大约90kDa处有一蛋白条带，与预期大小相同。人Neurotensin—R表达在细胞膜上。结论成功构建了pRluc—hNeurotensinl—R重组表达载体，建立了该质粒转染HEK293的细胞模型。可被用于检测与其它受体间的相互作用以及研究Neurotensinl—R介导的细胞内信号转导机制，这将有助于探究疾病的发病机制及开发新的药物靶点。
