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目的 (1)探讨中国人群 4 q35区 Eco R 片段长度多态性及中国人面肩肱型肌营养不良症 (FSHD)患者 4 q35区D4 Z4的缺失规律。 (2 )研究中国人 4 q35的不稳定性及 4 q- 10 q相互易位的规律及其与 FSHD的关系。 (3)研究基因突变与FSHD临床表型的关系 ,初步探讨其分子发病机制。 (4 )建立适合我国人群和民族特点的 FSHD基因诊断方法体系 ,为本病防治奠定基础。 方法 (1)采用 Eco R / H ind 双酶切g DNA+p13E- 11Southern杂交方法检测正常人群及 FSHD患者 4 q35区 Eco R / p13E- 11片段 ,并应用 Image分析软件计算分析片段大小。 (2 )采用 Eco R / Bln 双酶切 +p13E- 11Southern杂交方法检测和区分来自 10 q2 6的同源性片段。 (3)应用 Bgl / Bln 双酶切、Southern杂交方法检测 4 q-型和10 q-型 D4 Z4片段 ,应用剂量分析软件检测片段的杂交信号强度 ,计算 4 q∶ 10 q剂量比 ,推测 4 q- 10 q相互易位形式。采用 χ2检验比较正常人、散发型 FSHD和家族型 FSHD的易位率差异。 (4 )采用线性回归法分析患者发病年龄、丧失行走能力的年龄与 Eco R 片段长度的相关关系 ,采用 t检验比较易位和非易位组及不同性别患者的片段长度及发病年龄差异。 (5 )联合应用双酶切 +剂量检测方法进行基因诊断。 结果 (1)Eco R
Objective (1) To investigate the length polymorphism of Eco R fragment in 4 q35 region of Chinese population and the deletion of D4 Z4 gene in 4 q35 region of Chinese patients with surface-shoulder-muscular dystrophy (FSHD). (2) To investigate the instability of 435 and the reciprocal translocation of 4q-10q in Chinese and its relationship with FSHD. (3) To study the relationship between gene mutation and clinical phenotype of FSHD and to explore its molecular pathogenesis. (4) To establish a system of FSHD gene diagnostic method suitable for the Chinese population and ethnic characteristics, and lay a foundation for the prevention and treatment of this disease. Methods (1) The Eco R / p13E-11 fragment of 4 q35 region of normal population and FSHD patients was detected by Eco R / H ind double digestion with g DNA + p13E-11 Southern blotting. The fragment size was calculated by Image analysis software. (2) Homology fragments from 10 q2 6 were detected and distinguished by Eco R / Bln double digestion + p13E-11 Southern hybridization. (3) The 4 q-type and 10 q-type D4 Z4 fragments were detected by Bgl / Bln double digestion and Southern hybridization. The hybridization signal intensity of fragments was detected by the software of dose analysis. The dose ratio of 4 q: 10 q was calculated. q- 10 q translocation form. Chi-square test was used to compare the differences in translocation between normal and sporadic FSHD and familial FSHD. (4) The relationship between the age of onset and incapacity of walking and the length of Eco R fragment were analyzed by linear regression. The differences of fragment length and age of onset between translocation and non-translocation group and different gender were compared by t-test. (5) combined application of double digestion + dose detection method for gene diagnosis. Results (1) Eco R