先天性肛门直肠畸形合并少见型手足畸形一家系调查及其病因研究

 目的探讨先天性肛门直肠畸形合并少见型手足畸形一家系患者的临床表现及其致病原因。方法在患者家系调查的基础上，设立正常对照组与患者组，应用聚合酶链反应技术对候选基因P63第3-14，DLX5,DLX6,DAC,和HOXD13外显子的DNA序列测定，对两组的外显子进行分析。结果根据病史及体检提示，该家系患者祖孙三代均患有畸形程度进行性加重的肛门直肠畸形及手足畸形，其中2例肢体畸形符合缺指裂掌畸形；病因研究提示，将患者扩增的片段DNA序列分别与正常对照组以及人类基因库DNA序列进行比较，发现位于P63基因第6外显子的第797位碱基对发生突变，即由G突变为A，密码子CGA->CAA，该突变结果为错义突变，导致相对应的第227位氨基酸由精氨酸->谷氨酰胺。结论肛门直肠畸形合并缺指裂掌畸形可能是1个新的综合征表现型，由P63基因第6外显子基因组的第797位碱基对突变所致

Pedigree Investigation and Pathogeny Research on Congenital Anorectal Malformations Associated with Rare Limb Defects

Objective The paper is a To study on the clinical symptoms and pathogeny of ongenital anorectal malformations associated with limb defects of in some patients in from one family. Methods Based on the patients’ family pedigree investigation, a normal control group and a patient group were established. Then, DNA was extracted from leucocytes by the standard salt-precipitation method. Oligonucleotide primers were designed for exons 3-14 of P63, DLX5, DLX6, DAC and HOXD13 using Primer Premier 5.0, and direct sequence analysis of the exons (including exon-intron boundaries) was performed. Results The medical examination found that the patients manifested progressed ARMs and limb defects. The proband (IV1) and his father were diagnosed as SHFM. Through NCBI Blast and DNASTAR program, extensive sequencing analysis revealed a missense mutation in the DNA binding domain and at position located in exon 6 of P63: Arg227Gln (797G→A), (Fig.3) but which was not found in the five unaffected individuals in of the family and or the control group. Conclusion The congenital anorectal malformations associated with limb defects are caused by the P63 mutation of at number 797 base pair at number 797 of the exon 6 of P63.