Analysis of Clonality in Endocrine Neoplasms

利用荧光引物dna提取女性各种疾病患者外周血白细胞maoa微卫星法分析内分泌肿瘤的克隆性。采用荧光标记引物聚合酶链反应（PCR）扩增单胺氧化酶A型（MAOA）基因。扩增产物变性，在6%聚丙烯酰胺凝胶中电泳，用自动测序仪检测条带，使用GeneScan 672软件（Applied Biosystems）分析条带。接受检查的患者有；醛固酮产生性腺瘤：14例，库欣综合征（肾上腺腺瘤）：5例，嗜铬细胞瘤：1例，肾上腺阳刚化瘤：1例，甲状腺肿瘤：4例。所有24例患者的DNA均进行克隆分析。24例中有17例为MAOA基因杂合，杂合度为67%。人甲状旁腺裸鼠异体移植将手术切除的增生腺、腺瘤腺和正常腺制备成移植物，植入受体小鼠臀肌。通过测定小鼠血清中人完整甲状旁腺激素的浓度来评价移植物功能。甲状旁腺组织移植导致PTH浓度升高4周，此后达到平台期。8周内水平保持不变。血清PTH浓度与移植组织体积直接相关。移植甲状旁腺组织对血清低钙水平的反应也被观察到。

Analysis of Clonality in Endocrine Neoplasias-MAOA microsatelite methods using fluorescence primerDNA was extracted from peripheral white blood cells in female patients of various kinds of diseases. Monoamine oxidase type A (MAOA) gene was amplified by polymerase chain reaction (PCR) using fluorescence labeled primers. The amplified products were denatured and electrophoresed in 6% polyacrylamide gel and the bands were detected by automatic sequencer and analyzed using GeneScan 672 software (Applied Biosystems). The examined patients were ; aldosterone producing adenoma : 14, Cushing's syndrome (adrenal adenoma) : 5, pheochromocytoma : 1, virilizing adrenal tumor : 1, thyroid neoplasms : 4. All 24 patients' DNA were subjected to the clonal analysis. The informed cases (heterozygous for MAOA gene) were 17 of 24, so that the heterozygosity was 67%.Heterotransplantation of Human Parathyroid Glands into Nude MiceGrafts prepared from hyperplasia, adenoma, and normal glands which were resected at operation were implanted in the gluteus muscle of the recipient mice. Graft function was evaluated by measuring human intact PTH concentrations in sera of the mice. Transplantation of parathyroid tissues resulted in an increase in PTH concentrations for 4 weeks, reaching a plateau thereafter. The level remained unchanged for 8 weeks. Serum PTH concentrations were directly correlated with the volume of implanted tissues. The response of grafted parathyroid tissue to a low calcium level in sera was also observed.