Strategy of ATP7B gene analysis for Japanese patients with Wilson disease, using ARMS method. ..................................

1.日本肝豆状核变性和门克斯病患者的分子分析我们对40例日本肝豆状核变性患者的ATP7B基因和2例日本经典型门克斯病患者的ATP7A基因进行了分析。R778L、A874V和2871delC突变是日本肝豆状核变性患者常见的突变，占等位基因的50%以上。此外，我们还发现了孟克斯病患者2491insA的新突变。这位患者的母亲没有发生突变。因此，这种突变是从头开始的突变。R778L纯合子突变患者均为神经病学类型的肝豆状核变性。同时，所有携带2871delC突变纯合子的患者均表现出严重的肝功能障碍和/或进行性肝功能衰竭。我们推测R778L突变是神经表型的特异性突变，2871delC突变可导致严重的肝损害。对1例经典型Menkes病患者及其母亲、5例日本肝豆状核变性患者和2例正常人的淋巴母细胞系进行了铜诱导的ATPase活性测定。铜诱导的淋巴细胞ATPase活性丧失分别为29.1±4.2%(Menkes病)、48.2±1.9%(其母)和42.7~60.8%(Wilson病)。在本研究中，Menkes病患者、该病携带者和肝豆状核变性患者铜诱导的ATPase活性明显低于正常人。该方法可作为铜代谢紊乱的功能分析方法。

1. Molecular analysis for Wilson disease patients and Menkes disease patients in JapanWe analyzed ATP7B gene for 40 Japanese Wilson disease patients and ATP7A gene for 2 Japanese classical Menkes disease patients. R778L, A874V and 2871delC mutations were common mutations in Japanese Wilson disease patients.These 3 mutations could be found more than 50% of alleles. Also, we found novel mutation of Menkes disease patient, 2491insA. Mother of this patients had no mutation. Thus this mutation was de novo mutation.2. Genotype-phenotype correlation of Wilson diseaseAll of the patients who had R778L mutation homozygously were neurologic type of Wilson disease. Also all of patients who had 2871delC mutation homozygouly revealed severe liver dysfunction and/or progressive liver failure. We speculate that R778L mutation is specific for neurologic phenotype and 2871delC mutation causes svere hepatic damage.3. Copper specific P-type ATPase activityCopper-induced ATPase activities were measured relative to the lymphoblast cell lines derived from one case of classical Menkes disease patient and his mother, five Japanese patients with Wilson disease, and two normal subjects were investigated. Loss of copper-induced ATPase activities in lymphoblast were 29.1 ± 4.2% (Menkes disease patient), 48.2 ± 1.9% (his mother) and 42.7-60.8% (Wilson disease patients). In this study, copper-induced ATPase activities of Menkes disease patient, carrier of this disease and Wilson disease patients were definitely lower than that of normal subjects. This method will be useful as a functional analysis for copper metabolic disorders.