Mutation analysis of seven patients with Waardenburg syndrome北大核心CSCD

Objective: To perform genetic analysis for 7 patients with Waardenburg syndrome. Methods: Potential mutation of MITF, PAX3, SOX10 and SNAI2 genes was screened by polymerase chain reaction and direct sequencing. Functions of non-synonymous polymorphisms were predicted with Polyphen2 software. Results: Seven mutations, including c. 649-651delAGA (p. R217del), c. 72delG (p. G24fs), c. 185T>C (p. M62T), c. 118C>T (p. Q40X), c. 422T>C (p. L141P), c. 640C>T (p. R214X) and c. 28G>T (p. G43V), were detected in the patients. Among these, four mutations of the PAX3 gene (c. 72delG, c. 185T>C, c. 118C>T and c. 128G>T) and one SOX10 gene mutation (c. 422T>C) were not reported previously. Three non-synonymous SNPs (c. 185T>C, c. 128G>T and c. 422T>C) were predicted as harmful. Conclusion: Genetic mutations have been detected in all patients with Waardenburg syndrome. © 2016, West China University of Medical Sciences. All rights reserved.     

抗白灵霜在黑素合成中对TYR和MITF的调节作用

目的：明确抗白灵霜(KBL)在黑素合成中对TYR和MITF的调节作用。方法：体外培养小鼠B16黑素瘤细胞,分为空白对照组和不同浓度KBL（5 mg/mL,10mg/mL和15mg/mL）用NaOH溶解法检测黑素合成量,Real time-PCR检测TYR和MITF基因mRNA表达,Western Blot检测KBL组TYR和MITF蛋白的表达。结果：各浓度KBL组黑素合成量及TYR和MITF基因的mRNA和蛋白表达的水平均高于空白对照组,且呈浓度依赖性。结论：KBL可能通过上调MITF基因的表达,激活参与黑素合成的TYR以促黑素合成。     

补骨脂对SCF/Kit/MITF信号通路的调控作用

目的探讨中药单体补骨脂对SCF/Kit/MITF信号通路的调控作用。方法建立黑素细胞和角质形成细胞共培养体系,加入终浓度为2mg/mL的补骨脂醇提物,每24h更换新鲜培养基和补骨脂素1次,连续3d后收集细胞,RT-PCR法检测正常对照组和补骨脂组SCF,c-kit,MITF和酪氨酸酶mRNA的表达。用终浓度2mg/mL的补骨脂处理角质形成细胞爬片,连续3d后用免疫组化法检测正常对照组和补骨脂组SCF蛋白的表达。黑素细胞爬片,加入经补骨脂处理后的角质形成细胞上清,免疫组化法检测正常对照组和补骨脂组MITF蛋白的表达。结果补骨脂组SCF,c-kit,MITF和酪氨酸酶mRNA表达均较正常组明显增高(P=0.000)。经补骨脂处理后的角质形成细胞SCF免疫组化结果显示,细胞浆呈深棕黄色着色,与正常对照组的差异有统计学意义(P=0.000),加入经补骨脂处理的角质形成细胞培养上清后的黑素细胞MITF免疫组化结果显示,细胞核呈深棕黄色着色,与正常培养黑素细胞的差异有统计学意义(P=0.007)。结论中药单体补骨脂能促进角质形成细胞分泌SCF,进而促进MITF的表达,使酪氨酸酶表达增加。