血友病是凝血因子Ⅷ/Ⅸ基因突变引起的性连锁隐性遗传性出血性疾病，致残率及死亡率高，需终生输注凝血因子，药物治疗尚属空白。凝血因子基因中产生提前终止的无义密码子(PTC)，启动无义密码子介导的mRNA降解（NMD），导致凝血因子的表达质/量降低，引起重型血友病的发生。由于目前在我国血友病中无义突变的比例尚不明确,本课题拟采用PCR和直接测序进一步筛查血友病A/B患者所有外显子的无义突变，明确无义突变在血友病发病中的比例；分离携带无义突变家系成员骨髓间充质干细胞，诱导分化为肝样细胞，检测其FVIII/FIX的含量及活性，进行不同浓度非氨基糖苷类小分子化合物（PTC124）药物干预，检测药物干预前后FⅧ/FⅨ的 mRNA、蛋白表达量及活性变化，为血友病的发病机制和药物治疗奠定基础。

The hemophilia, which is caused by the mutation in the factor Ⅷ/Ⅸ (FⅧ/FⅨ)gene, is known as an X-linked recessive disease with high disability rate and mortality. At present, due to lacking of the eradicative therapy, the patients needed infusion the expensive blood coagulation factor in their lifetime. One of the most important molecular mechanisms in severe hemophilia is nonsense mutation, carrying premature termination codon(PTC),which lead to the production of nonfunctional truncated protein or the degradation of PTC-bearing mRNA.Because the percentage of nonsense mutation in hemophilia in our country is not clear, we try to use PCR and direct DNA sequencing to screen and identify the percentage of nonsence mutation in patients with hemophilia A and B. Furthermore,isolating and culturing the bone marrow mesenchyme stem cells of the patient's family members with nonsense mutation, induced bone marrow mesenchyme stem cells differentiate into hepatocyte-like cells and detected the FⅧ/FⅨ protein level and activities. Then, after treated with different concentration of chemical compound PTC124, testing the variations of FⅧ/FⅨ mRNA and protein quantities and activities,further research the mechanism and treatment of hemophilia.