脊髓性肌萎缩症(Spinal muscular atrophy,SMA)大多数在儿童或婴幼儿期发病,表现为进行性、对称性的肢体无力和肌肉萎缩,迄今尚无有效的治疗方法,是婴幼儿最常见的致死性遗传病之一。患者来源的细胞系是该病研究的重要工具,但依赖于肌...脊髓性肌萎缩症(Spinal muscular atrophy,SMA)大多数在儿童或婴幼儿期发病,表现为进行性、对称性的肢体无力和肌肉萎缩,迄今尚无有效的治疗方法,是婴幼儿最常见的致死性遗传病之一。患者来源的细胞系是该病研究的重要工具,但依赖于肌肉或皮肤活检等创伤性手术的成纤维细胞培养较难被患者及家属接受。文章收集SMA患者及健康对照的新鲜尿液,进行离心、尿液沉渣培养,观察尿液细胞的生长状况,用酶联免疫吸附实验(Enzyme-linked immunosorbent assay,ELISA)分析患者尿液细胞中SMN(Survival of motor neuron)蛋白的表达量,应用免疫荧光染色观察SMN蛋白在细胞内的定位。共建立了11例SMA患者和14例健康对照的尿液细胞系,尿液细胞体外增殖旺盛,细胞形态及生长速度较稳定。患者来源的尿液细胞SMN1(Survival of motor neuron 1)基因缺失突变、SMN蛋白表达量降低,荧光染色提示SMN蛋白在胞浆和胞核中均有定位。尿液细胞培养步骤简单、无创伤性、患儿及其家属的依从性好,是获取和保存病人来源标本的有效方法,在脊髓性肌萎缩症发病机制研究和临床应用方面具有较好的应用价值。展开更多
Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three...Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are rec- ognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMN1 and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMA1 patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NAIP gene may be a modifying factor for disease severity of SMA1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA.展开更多
文摘脊髓性肌萎缩症(Spinal muscular atrophy,SMA)大多数在儿童或婴幼儿期发病,表现为进行性、对称性的肢体无力和肌肉萎缩,迄今尚无有效的治疗方法,是婴幼儿最常见的致死性遗传病之一。患者来源的细胞系是该病研究的重要工具,但依赖于肌肉或皮肤活检等创伤性手术的成纤维细胞培养较难被患者及家属接受。文章收集SMA患者及健康对照的新鲜尿液,进行离心、尿液沉渣培养,观察尿液细胞的生长状况,用酶联免疫吸附实验(Enzyme-linked immunosorbent assay,ELISA)分析患者尿液细胞中SMN(Survival of motor neuron)蛋白的表达量,应用免疫荧光染色观察SMN蛋白在细胞内的定位。共建立了11例SMA患者和14例健康对照的尿液细胞系,尿液细胞体外增殖旺盛,细胞形态及生长速度较稳定。患者来源的尿液细胞SMN1(Survival of motor neuron 1)基因缺失突变、SMN蛋白表达量降低,荧光染色提示SMN蛋白在胞浆和胞核中均有定位。尿液细胞培养步骤简单、无创伤性、患儿及其家属的依从性好,是获取和保存病人来源标本的有效方法,在脊髓性肌萎缩症发病机制研究和临床应用方面具有较好的应用价值。
基金Project supported by the National Natural Science Foundation of China (No. J0710043)the Natural Science Foundation of Zheji-ang Province (No. 2007C33049), China
文摘Spinal muscular atrophy (SMA) is a disorder characterized by degeneration of lower motor neurons and occasionally bulbar motor neurons leading to progressive limb and trunk paralysis as well as muscular atrophy. Three types of SMA are rec- ognized depending on the age of onset, the maximum muscular activity achieved, and survivorship: SMA1, SMA2, and SMA3. The survival of motor neuron (SMN) gene has been identified as an SMA determining gene, whereas the neuronal apoptosis inhibitory protein (NAIP) gene is considered to be a modifying factor of the severity of SMA. The main objective of this study was to analyze the deletion of SMN1 and NAIP genes in southern Chinese children with SMA. Here, polymerase chain reaction (PCR) combined with restriction fragment length polymorphism (RFLP) was performed to detect the deletion of both exon 7 and exon 8 of SMN1 and exon 5 of NAIP in 62 southern Chinese children with strongly suspected clinical symptoms of SMA. All the 32 SMA1 patients and 76% (13/17) of SMA2 patients showed homozygous deletions for exon 7 and exon 8, and all the 13 SMA3 patients showed single deletion of SMN1 exon 7 along with 24% (4/17) of SMA2 patients. Eleven out of 32 (34%) SMA1 patients showed NAIP deletion, and none of SMA2 and SMA3 patients was found to have NAIP deletion. The findings of homozygous deletions of exon 7 and/or exon 8 of SMN1 gene confirmed the diagnosis of SMA, and suggested that the deletion of SMN1 exon 7 is a major cause of SMA in southern Chinese children, and that the NAIP gene may be a modifying factor for disease severity of SMA1. The molecular diagnosis system based on PCR-RFLP analysis can conveniently be applied in the clinical testing, genetic counseling, prenatal diagnosis and preimplantation genetic diagnosis of SMA.
