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在数据分析之前,我们先用Shapiro-Wilk法对数据的分布类型进行分析。结果显示:0.0, 0.5, 1.0, 1.5, 2.0 和2.5 Gy辐射剂量点的原始数据均服从正态分布, P值分别是0.260, 0.072, 0.390, 0.100, 0.250和0.290,均大于0.050拒绝零假设( H 0),服从正态分布。随后使用方差分析得出 F=39.65; P<0.000 1,提示各组之间MNF有差异性,要深入研究具体组别之间的差异性,使用SNK法得出每两组之间均有显著差异性( P<0.000 1),并且随着辐照剂量的增加MNF也逐渐增加。此结果与文献报道一致 [ 24- 25] ,MNF诱导呈剂量依赖性,每一个样本的MNF都随着辐照剂量的增加而增加[ 图1(b)],剂量效应曲线均符合线性平方模型;但是,相同剂量辐照不同样本诱导的MNF存在差异[ 图1(b)],体现出个体辐射敏感性的差异。以50个样本的MNF平均值及其分散程度,拟合剂量效应二阶多项的中心方程为MNF=0.014 7+0.036 2D+0.023 1D 2 [ 图1(c),蓝色线条],回归系数 r为0.726。
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个体辐射敏感性的界定目前国内外都还没有标准。参照辐射损伤实验研究和临床肿瘤放射治疗的惯例,一般可以通过比较相同辐照剂量(例如2 Gy)诱导的生物效应的高低,或者达到相同的生物学终点需要的辐射剂量大小两种方法来区分敏感性,相同辐照剂量诱导的生物效应高的辐射敏感性高,或者达到相同的生物学终点需要的辐射剂量小的辐射敏感性高。两种方法均是采用单个剂量点来进行评价,往往需要通过足够的重复实验次数来获得可靠的实验结果。但是,考虑实际工作中反复抽取被试者的血液可操作性低,每个样本(即每位志愿者)MNF的测定很难进行三次以上的有效重复,为了降低单次实验的误差,我们测定了每个样本6个辐照剂量点的微核数据,据此绘制辐射诱导微核的剂量效应曲线,“以线代点”,以量效关系来确定个体辐射敏感性。然后,以50个样本每个剂量诱导的MNF平均值来绘制标准剂量效应曲线。最后,将每个样本的剂量效应曲线与标准剂量效应曲线进行比对,所有5个剂量点(不含本底)的MNF均高于平均值的样本,定义为辐射敏感人(群);每个样本所有5个剂量点的MNF均低于平均值的定义为辐射抗性人(群);5个剂量点中有的高于平均值、有的低于或者等于平均值的,定义为普通人(群),具体示例如 图2(a)所示;随后,敏感人群的MNF均值、抗性人群MNF均值分别与50位志愿者均值进行统计学分析[ 图2(b)],分析结果表明:在各个剂量点下,敏感人群、抗性人群均与50位志愿者之间存在MNF差别,具有统计学差异。依据此标准我们发现,50个样本中对辐射敏感的有13人(占26%)、对辐射抗性的14人(占28%),基本符合正态分布(见 表1)。
表 1 个体辐射敏感性的分布
背景微核率 合计人数 敏感人数 普通人数 抗性人数 敏感人群比例/% 普通人群比例/% 抗性人群比例/% 全部志愿者 50 13 23 14 26.0 46.0 28.0 高 13 6 6 1 46.2 46.2 7.6 中 22 6 11 5 27.3 50.0 22.7 低 15 1 6 8 6.7 40.0 53.3 -
按照3.2节划分个体辐射敏感性的方法我们将人群分为辐射敏感人群、普通人群和抵抗人群( 表1)。15位低本底的志愿者中仅1人(6.7%)对辐射敏感,却有8人(53.3%)对辐射抗性;13位高本底的自愿者中6人(46.2%)对辐射敏感,仅1人(7.6%)对辐射抗性。因此,我们分析个体敏感程度与本底MNF之间的相关性,由于数据为等级变量,我们采用等级变量的秩和相关分析来验证两个变量之间的相关性。结果发现:本底MNF和辐射敏感性呈正相关,即本底MNF低的样本中辐射敏感性人数少、抗性人群多;本底MNF高的样本中辐射敏感人数多、抗性人群少( 表1)。为了研究本底MNF与不同辐射后各个辐射剂量点MNF之间的相关性,使用线性相关做散点图并且进行拟合,结果如 图3所示,0.5, 1.0, 1.5, 2.0 和2.5 Gy分别照射后的MNF与本底MNF之间均呈现正相关。在各个辐射剂量点的相关系数 r分别是0.738 9, 0.654 9, 0.616 5, 0.662 6和0.601 3,并且 P值均小于0.05,说明在每个辐射剂量点均呈现出本底MNF越高则辐射敏感性越高的趋势。
Preliminary Study on Male Individual Radiosensitivity by Cytochalasin-B Blocked Micronucleus Assay
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摘要: 初步研究了男性个体辐射敏感性的鉴定方法及标准。采集50名男性志愿者的外周血,分别给予不同剂量X射线照射,采用细胞松弛素B阻断双核法测定微核率(MNF),通过二阶多项拟合法,绘制微核剂量效应选项中心标准曲线,将个人微核剂量效应曲线与标准曲线比对后判断个体辐射敏感性。
$0.0\sim2.5 $ Gy剂量范围内,剂量效应二阶多项拟合的中心方程为(MNF=0.014 7+0.036 2D+0.023 1D 2, r=0.726)。50名志愿者中,辐射敏感的有13人,辐射抗性的有14人,基本符合正态分布。Spearman秩和相关分析结果显示,MNF在各个辐射剂量点与辐射敏感性均存在正相关,与辐射抗性呈负相关,MNF随剂量增加而增加。本研究初步建立了“以线代点”男性个体辐射敏感性鉴定方法,并发现男性外周血淋巴细胞的本底微核率与个体辐射敏感性呈正相关。Abstract: Methods and criteria for the identification of male individual radiosensitivity are investigated. The peripheral blood of 50 male volunteers was irradiated with different doses of X-ray, the micronucleus frequency(MNF) was measured by cytokinesis-B block micronucleus assay, and the central standard curve of micronucleus dose effect was drawn by second-order multinomial fitting method so that the individual dose-response curve of micronucleus was compared with the standard curve to judge individual radiosensitivity. In the dose range of 0.0~2.5 Gy, the central equation of second-order polynomial fit for dose effect is (MNF=0.014 7+0.036 2D+0.023 1D 2, R=0.726). Among the 50 male volunteers, 13 were sensitive to radiation (26.0%) and 14 were resistant (28.0%), which basically fits the normal distribution. Spearman rank correlation analysis showed that the MNF increased with the dose and there was a positive correlation between the background MNF and the radiosensitivity of each dose, but negative correlation with the radioresistance. In conclusion, a "curve-instead-of-dose" male individual radiosensitivity identification method was established. Moreover, it was found that the background of micronuclei in peripheral blood lymphocytes was positively correlated with individual radiosensitivity.-
Key words:
- male individual radiosensitivity /
- ionizing radiation /
- micronucleus /
- dose effect
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表 1 个体辐射敏感性的分布
背景微核率 合计人数 敏感人数 普通人数 抗性人数 敏感人群比例/% 普通人群比例/% 抗性人群比例/% 全部志愿者 50 13 23 14 26.0 46.0 28.0 高 13 6 6 1 46.2 46.2 7.6 中 22 6 11 5 27.3 50.0 22.7 低 15 1 6 8 6.7 40.0 53.3 -
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