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应用胞浆分裂阻断微核法,以抗着丝粒血清标记微核( M N),针对血清各种微量营养素对两种自发染色体损伤,即整条染色体丢失和染色体断裂的影响进行了研究。方法:选择 33 名 19~69 岁未使用过任何已知和可疑诱变剂、在此之前 8 个月内未进行过 X 射线检查的健康非吸烟者。取空腹血分离淋巴细胞进行培养,44 小时时加松胞素 B,终浓度为 30μg/m l,培养至 72 小时,经细胞涂片、干燥、固定并染色。将 Tris 缓冲液按 1/8 稀释的抗着丝粒血清加入到固定了细胞的玻片上,于 5℃湿盒内孵育过夜,冲洗后加入过氧化物酶标记的兔抗人 Ig G,于室温孵育 3 小时。用改进的氯化镍/咪唑标准二氨基联苯胺反应进行组化检查。甲基绿复染。计数1 000 个双核细胞确定微核细胞( M N C)率。至少计数 27 个 M N 确定着丝粒(+ )和(- )比例。血清从全血中分离,测量其中β胡萝卜素、维生素 B12 、维生素 C、叶酸和维生素 E等微量营养素含量。结果:简单相关分析表明,年龄与 M N C 率的增加显著相关。与年龄相关增加的 M N C 既包含着丝粒(+ ),也包含着丝粒(- )的 M N。除维生素 B12与着丝粒(+ ) M N C 率呈弱正?
The effects of serum micronutrients on two kinds of spontaneous chromosomal damage, namely the whole chromosome loss and chromosome rupture, were studied by applying cytoskeletal blockage micronuclei method and anti-centromere-labeled micronuclei (M N) . METHODS: Twenty-three healthy non-smokers, aged 19-69 who had not used any known and suspected mutagens, and who had not had an X-ray examination within the preceding 8 months, were enrolled. The fasting blood was collected for lymphocyte separation. Cytokinin B was added 44 hours later, and the final concentration was 3 0μg / ml. The cells were cultured to 72 hours. The cells were smeared, dried, fixed and stained. The Tris buffer was diluted 1/8 with anti-centromeric serum to the cell-mounted slide and incubated overnight at 5 ° C in a wet box. After washing, peroxidase-labeled rabbit anti-human Ig G was added and incubated at room temperature Incubate for 3 hours. Histochemical examination was performed with the improved nickel chloride / imidazole standard diaminobenzidine reaction. Methyl Green counterstain. Counting 1,000 dinuclear cells determines the rate of micronuclei (MNC). Count at least 27 M N to determine the centromere (+) and (-) ratio. Serum was separated from whole blood and the levels of micronutrients such as β-carotene, vitamin B12, vitamin C, folic acid and vitamin E were measured. Results: Simple correlation analysis showed that age was significantly associated with an increase in M N C rate. The age-dependent increase of MNC includes both centromere (+) and centromere (-) MN. In addition to vitamin B12 and centromere (+) M N C rate was weak positive?