冠状动脉CT成像不同管电压对外周血淋巴细胞DNA双链断裂影响的研究

【摘要】目的：利用流式细胞仪评价冠状动脉CT成像（CCTA）不同管电压对外周血淋巴细胞DNA双链断裂（DSBs）水平的影响。方法：将120例行CCTA检查的患者随机分为A、B、C、D四组,分别接受管电压为120kV、100kV、80kV、70kV的CCTA扫描。在CCTA检查前及检查后20min分别抽取外周静脉血2mL,符合标准的血样经过淋巴细胞分离及抗γ-H2AX蛋白和抗ATM蛋白抗体孵育后,利用流式细胞仪分析T淋巴细胞内DNA双链断裂水平。对CCTA检查前、后DNA双链断裂的改变情况及不同管电压组间DNA的损伤情况进行统计学分析。结果：四组CCTA扫描后γ-H2AX和ATM的阳性细胞百分比均有不同程度增加（P＜0.05）。管电压由120kV降低至80kV,γ-H2AX阳性细胞百分比的差值逐渐减小;管电压降低至70kV,γ-H2AX阳性细胞百分比的差值增高;组间比较发现,除A组与C组（P=0.007）、B组与C组之间（P=0.003）差异有统计学意义外,其余组间差异均无统计学意义（P值均＞0.05）。随着管电压的降低,四组间ATM阳性细胞百分比差值的差异无统计学意义（P=0.381）。结论：经流式细胞仪检测发现,CCTA管电压降低至80kV能够有效减少DNA双链断裂。     

γ-H2AX分析用于检测电离辐射致HepG2细胞DNA双链断裂的研究

目的 应用γ-H2AX分析检测电离辐射导致肝癌细胞株HepG2基因组DNA双链断裂的情况，并检测在不同细胞周期时相中的表达差异，从而了解不同时相HepG2细胞株的辐射敏感性。方法利用胸腺嘧啶核苷（TdR）阻断HepG2细胞于G1期末，于不同时间点分别得到同步化的S期和G2/M期细胞，用60Coγ射线照射细胞，建立DNA双链断裂模型，采用免疫荧光法和Westemblotting检测γ-H2AX的表达。结果TdR阻断后继续培养至34和40h，分别得到了较高同步化程度的S期和G2／M期HepG2细胞；受照后的HepG2细胞中Y-H2AX表达较照射前显著增高，处于S期的细胞γ-H2AX表达增加更为突出。结论不同周期时相的HepG2细胞受照后均检测出不同程度的DNA双链断裂，其中S期细胞尤为敏感。γ-H2AX对DNA双链断裂快速敏感的反应使γ-H2AX分析在检测早期DNA双链损伤中具有广泛的廊用前景。     

γ-H2AX分析电离辐射诱发小鼠神经元DNA双链断裂及修复

目的 观察临床剂量的电离辐射后小鼠神经元DNA双链的断裂及修复, 探讨γ-H2AX是否能作为衡量体内正常脑组织神经元DNA双链断裂(DSB)形成和修复的指标。方法 电离辐射诱导DSB形成试验,C57BL/6小鼠行0.1、0.5和1.0 Gy全身照射后10 min,收集脑组织进行分析;DSB修复试验,修复功能正常小鼠(C57BL/6)和修复缺陷鼠(BALB/c, A-T 和SCID)在全身照射2 Gy后0.5、2.5、5、24和48 h收集脑组织进行分析。未照射的小鼠作为对照组。γ-H2AX和NeuN免疫荧光双重染色和免疫组织化学染色分析脑组织神经元DSB形成和修复。结果 DSB形成试验,在对照组脑组织皮质区神经元的细胞核内仅有数目很少的γ-H2AX焦点,而受照射后细胞核内γ-H2AX焦点数目显著增加,并显示出明显的剂量相关。通过分析不同放射敏感性的小鼠电离辐射后脑组织皮质区神经元的DSB修复动力学,发现C57BL/6小鼠细胞核内γ-H2AX焦点随时间的延长迅速减少,在照射后24和48 h仅有很低水平DSB未修复;而免疫缺陷SCID鼠在照射后所有的时间点都显示出γ-H2AX焦点的明显增加,A-T小鼠表现出较低的修复缺陷,主要表现在较晚的时间点(≥5 h)γ-H2AX焦点的中度增加;放射敏感的BALB/c小鼠与C57BL/6小鼠相比γ-H2AX焦点数量轻度增加。结论 γ-H2AX焦点分析可以作为一项精确的量化指标,在体内衡量临床相关剂量电离辐射诱导的DSB形成和修复。     

冠状动脉多层CTA评价冠状动脉支架的最新进展：双源CT体外评估29种不同支架

本研究的目的是应用双源多层CT4种图像重建法测试体外大样本最新冠状动脉支架的管腔显示、管腔衰减值及图像噪声,并提供目前常用冠状动脉支架的CT目录。29种不同的冠状动脉支架（19种钢制、6种钴铬合金、2种钽制、1种铁制、1种镁制）均置于冠状动脉模型（血管直径3mm,血管衰减值250HU,血管外密度-70HU）中检查。     

Tip60基因沉默对细胞DNA双链断裂修复和辐射敏感性的影响

目的 探讨Tip60对细胞辐射敏感性的影响及相关机制。方法 采用siRNA和Tip60乙酰转移酶抑制剂漆树酸,抑制U2OS细胞中Tip60的表达或乙酰转移酶活性;用克隆形成率分析细胞对⁶⁰Co γ射线的敏感性;采用γ-H2AX原位免疫荧光集簇点法,检测DNA双链断裂损伤修复;用免疫共沉淀检测蛋白质的相互作用。结果 siRNA沉默Tip60表达明显提高了U2OS细胞对1、2 Gy中、低剂量γ射线的敏感性(t=3.364、3.979,P＜0.05),但对4 Gy大剂量照射的细胞存活率无明显影响。γ-H2AX集簇点检测结果表明,照射后1、4和8 h,Tip60失活导致细胞DNA双链断裂修复能力降低(t=3.875、3.183和3.175, P<0.05)。细胞在受到电离辐射损伤后,Tip60与DNA修复蛋白DNA-PKcs发生相互作用,漆树酸能抑制DNA-PKcs的T2609位点的磷酸化。结论 Tip60通过与DNA-PKcs相互作用,调控细胞DNA双链断裂修复机制,对细胞辐射敏感性产生影响。     

基于DNA双链断裂损伤簇的相对生物效能模拟计算

目的从追溯辐射引起的DNA损伤机制出发, 根据基本的生物物理原理获取DNA双链断裂(DSB)损伤簇表示的相对生物效能(RBE)值, 探讨DNA损伤与染色体畸变、生殖细胞死亡生物效能的相似性。方法以低能电子、质子、α粒子作为研究对象, 通过辐射物理化学模型, 模拟细胞核受粒子辐射的过程。在基于损伤点密度的含噪声密度聚类(DBSCAN)算法的基础上, 改进DSB损伤簇分类方法, 以减弱模拟过程中损伤点与能量沉积随机分布假设的联系, 使得DSB损伤簇更接近于非随机分布。计算获得DSB损伤簇的产额, 提出以DSB损伤簇计算RBE值的方法。结果 2 MeV α粒子的DSB损伤簇RBE值(12.29)与生物实验中染色体片段(15.3±5.9)、细胞存活(14.7±5.1)表示的RBE相近。结论高传能线密度(LET)电离辐射后, 不同于简单DSB, DSB损伤簇的RBE与染色体畸变、细胞存活生物效能存在一定的相似性。     

低剂量γ射线诱导的淋巴细胞DNA断裂的适应性反应

利用ＤＮＡ解旋荧光测定法（ＦＡＤＵ）研究了低剂量γ射线诱导的淋巴细胞ＤＮＡ断裂的适应性反应，并观察了不同浓度的３－氨基苯甲酰胺（３－ＡＢ）对适应性反应的影响。结果表明：０．５～４．０ｃＧｙγ射线照射（Ｄ１），均可诱导淋巴细胞对１５Ｇｙγ射线照射的抗性，８．０ｃＧｙ引起的适应性反应不明显，２．０，４．０ｃＧｙ的γ射线为诱导适应性反应的最适剂量。５～２０Ｇｙ的Ｄ２照射均可显现出Ｄ１（２．０ｃＧｙ）诱导的适应性反应，以１５Ｇｙ照射后适应性反应最强。０．５ｍｍｏｌ／Ｌ的３－ＡＢ就可明显地抑制适应性反应。当浓度增至５．０ｍｍｏｌ／Ｌ时，适应性反应几乎完全受抑。     

基于随机森林的电离辐射诱导DNA双链断裂分类模型的构建与应用

目的 构建预测电离辐射诱导DNA双链断裂（DSB）水平的随机森林分类模型,初步研究DSB在基因组中的分布规律。方法 将GRCh38参考基因组分为50 kb的片段,根据MCF-7细胞的测序数据把片段分为电离辐射诱导的DSB低水平和高水平区域,以8种表观遗传学特征作为输入,随机将数据集的2/3列为训练集,1/3列为测试集,构建含100棵决策树的随机森林分类模型。分析分类模型中表观遗传学的特征重要性,展示这些标记在不同DSB水平区域的富集差异。结果 随机森林分类模型在测试集上预测的准确率为99.4%,精准率为98.9%,召回率为99.9%,受试者操作特征曲线下面积为0.994。8个特征中H3K36me3和DNase标记的重要性最高,富集分析表明DSB高水平区域的这两类标记明显高于DSB低水平区域。结论 以表观遗传学数据作为特征输入,随机森林分类模型可在50 kb基因组区域上准确预测电离辐射诱导的DSB水平,分析表明这些DSB可能主要分布在基因组中转录活跃的部位。     

二代双源CT对冠状动脉支架的显示及辐射剂量的研究

目的 对比分析二代双源CT前瞻性心电触发高螺距螺旋扫描模式(Flash模式)和回顾性心电门控螺旋扫描模式(常规模式)下冠状动脉支架的显示质量和辐射剂量.方法 常规模式和Flash模式下共计120例患者的155个冠状动脉植入支架纳入本研究,平均年龄(64.9±10.6)岁,所有患者的心率均控制在≤65次/min,且心律规则.所有患者分成两组,分别接受Flash模式和常规模式的冠状动脉成像.冠状动脉原始图像均进行Kernel B26和Kernel B46的数据重建,并由两位医师对支架的显示质量进行独立的4级评分.两种模式下的冠状动脉显示质量利用卡方检验进行统计分析,而不同冠状动脉扫描模式下的容积CT剂量指数值(CTDIvol)和剂量长度乘积值(DLP),利用t检验进行辐射剂量的对比研究.结果 2位医师对所有冠状动脉支架显示质量的评分具有较好的一致性(Kappa=0.764,P＜0.001),Flash模式和常规模式的平均评分分别为:1.61±0.77和1.65±0.82.两种冠状动脉扫描模式的图像质量差异无统计学意义(x2=0.865,P=0.834).Flash模式的CTDIvol(3.24±1.21)明显低于常规模式(31.26±10.79),差异有统计学意义(t=19.83,P<0.001);Flash模式的DLP(54.61±19.88)同样低于常规模式(468.30±174.88),差异有统计学意义(t=18.06,P＜0.001).结论 控制患者心率≤65次/min且心律规则,Flash模式在降低辐射剂量的前提下可获得与常规模式相同质量的冠状动脉支架图像.

Abstract：

Objective To compare a prospective ECG-gated high-pitch spiral technique (Flash) and conventional retrospective ECG-gated spiral technique for the image quality of coronary artery stent and radiation dose with a dual source CT.Methods One hundred and fifty five coronary stents in one hundred and twenty patients (mean age 64.9 ± 10.6 years,heart rates≤65 bpm) were examined using a dual source CT.All patients were divided in two groups,receiving either Flash or conventional coronary artery CT angiography separately.After images of coronary artery were reconstructed using both the smooth (B26) and sharp (B46) kernel,the coronary stent image quality and stent lumen were scored by two observers individually using four point scale (1 = excellent,4 = unvaluable) .The effective radiation dose of volume CT dose index (CTDIvol,mGy) and dose length product (DLP,mGy x cm) were also calculated for each patient.x2-test analysis of image quality and t-test analysis of radiation dose were used respectively for statistical difference between two groups.Results Interobserver agreement for stent image quality was good (Kappa =0.764,P＜0.001).The mean scores were 1.61 ±0.77 and 1.65 ±0.82 in Flash group and conventional group respectively.There was no significant difference in image quality between the two groups (x2 = 0.865,P = 0.834).The effective radiation dose in Flash group was significantly lower than that in conventional group.The mean values of CTDIvol were 3.24 ± 1.21 in Flash group and 31.26 ± 10.79 in conventional group (t = 19.83,P < 0.001) ,and the mean values of DLP in Flash group and conventional group were 54.61 ±19.88 and 468.30 ± 174.88,respectively (t = 18.06,P < 0.001).Conclusions Compared with the conventiaonal coronary artery CT angiography,the Flash coronary artery CT angiography technique has a similar coronary stent image quality,but at a lower radiation dose in patients with heart rates lower than 65 beats per minute.     

DNA双链断裂-潜在的指标,用于判断心脏CT和常规冠状动脉造影电离辐射产生的生物学效应:一项随机、对照研究

摘要目的前瞻性比较由心脏CT检查和常规冠状动脉造影(CCA)诱导的DNA双链断裂。方法56例疑似为冠心病的病人随机接受CCA或者心脏CT检查。采用血液淋巴细胞荧光显微镜的方法评估DNA双链断裂,将其作为反映辐射暴露的生物学效应指标。辐射剂量用剂量长度乘积(DLP)和剂量面积乘积(DAP),以及CT和CCA各自的转换因子来估算。结果CT组和CCA组每个淋巴细胞平均有0.12±0.06和0.29±0.18个诱导的DNA双链断裂(P<0.001)。     

Effect of CT scan protocols on x-ray-induced DNA double-strand breaks in blood lymphocytes of patients undergoing coronary CT angiography

Aims  

To compare in vivo DNA lesions induced during helical and sequential coronary computed tomography angiography (CTA) and to evaluate the effect of CT parameters on double-strand break (DSB) levels.     

Induction and repair of DNA double-strand breaks in blood lymphocytes of patients undergoing 18F-FDG PET/CT examinations

Purpose

The purpose of this study was to evaluate DNA double-strand breaks (DSBs) in blood lymphocytes of patients undergoing positron emission tomography (PET)/CT using γ-H2AX immunofluorescence microscopy and to differentiate between ¹⁸F-fluorodeoxyglucose (FDG) and CT-induced DNA lesions.

Methods

This study was approved by the local Ethics Committee and complies with Health Insurance Portability and Accountability Act (HIPAA) requirements. After written informed consent was obtained, 33 patients underwent whole-body ¹⁸F-FDG PET/CT (3?MBq/kg body weight, 170/100 reference mAs at 120?kV). The FDG PET and CT portions were performed as an initial CT immediately followed by the PET. Blood samples were obtained before, at various time points following ¹⁸F-FDG application and up to 24?h after the CT scan. Distinct foci representing DSBs were quantified in isolated lymphocytes using fluorescence microscopy after staining against the phosphorylated histone variant γ-H2AX.

Results

The DSB values at the various time points were significantly different (p?<?0.001). The median baseline level was 0.08/cell (range 0.06–0.12/cell). Peaks of radiation-induced DSBs were found 30?min after ¹⁸F-FDG administration (median excess foci 0.11/cell, range 0.06–0.27/cell) and 5?min after CT (median excess foci 0.17/cell, range 0.05–0.54/cell). A significant correlation between CT-induced DSBs and dose length product was obtained (ρ?=?0.898, p?<?0.001). After 24?h DSB values were still slightly but significantly elevated (median foci 0.11/cell, range 0.10–0.14/cell, p?=?0.003) compared to pre-exposure levels.

Conclusion

PET/CT-induced DSBs can be monitored using γ-H2AX immunofluorescence microscopy. Peak values may be obtained 30?min after ¹⁸F-FDG injection and 5?min after CT. The radionuclide contributes considerably to the total DSB induction in this setting.     

X-ray induced DNA double-strand breaks in coronary CT angiography: comparison of sequential, low-pitch helical and high-pitch helical data acquisition

Background

Aim of this study was to compare DNA double-strand breaks (DSBs) in blood lymphocytes of patients undergoing high-pitch helical, low-pitch helical and sequential coronary CT angiography.

Methods and results

66 patients were examined with various scan protocols and modes (low-pitch helical scan: 100–120 kV, 320–438 mAs/rot, pitch 0.18–0.39, with or without ECG-pulsing, n = 35; prospectively ECG-triggered high-pitch helical scan: 100–120 kV, 320–456 mAs/rotation, pitch 3.2–3.4, n = 19; prospectively ECG-triggered sequential scan: 100–120 kV, 150–300 mAs or 320–370 mAs/rotation, n = 12) either using a 64-slice or 128-slice dual-source CT or a 128-slice single source CT scanner. Blood samples were obtained before and 30 min after CT and DSBs were analyzed in isolated lymphocytes using γ-H2AX immunofluorescence microscopy.A significant increase of DSBs was measurable 30 min after CTA (range 0.01–0.71/cell). CT induced DSBs showed a significant correlation with the estimated effective dose (ρ = 0.90, p < 0.00001). Both prospectively ECG-triggered sequential (0.10 DSBs/cell, 176 mGy cm, p < 0.00001) and high-pitch helical scan protocols (0.03 DSBs/cell, 109 mGy cm, p < 0.00001) led to a significant reduction of median DLP and DSB levels compared to low-pitch helical scans (0.34 DSBs/cell, 828 mGy cm). A reduction of the tube voltage resulted in significantly lower whereas additional calcium scoring resulted in elevated DLP and DNA damages (p < 0.05 each).

Conclusion

In coronary CTA, data acquisition protocols have a significant influence on the X-ray induced DSB levels. Using γ-H2AX immunofluorescence microscopy different scan modes in different CT generations can be compared concerning their biological impact.     

在双源CT冠状动脉成像中应用不同低剂量扫描方案对影像质量和辐射剂量的影响:随机研究

目的比较在128层双源CT冠状动脉成像中应用不同低剂量扫描方案的图像质量、辐射剂量和它们与心率的关系。材料与方法此研究通过当地伦理委员会审查,所有病人都知情同意。272例病人(男175例,女97例,平均年龄分     

Effect of calcium scoring scan in reducing scan range and radiation dose in retrospectively ECG-gated CT coronary angiography

目的:探讨使用钙化积分扫描缩短增强扫描范围以降低后门控冠状动脉CTA检查辐射量的临床应用价值.方法:40例患者行冠脉钙化积分及冠脉CTA检查,其增强扫描的实际扫描范围根据钙化积分扫描图像来决定,即冠状动脉树上缘1 cm至心尖部心包影即将消失的层面.并按常规方法在定位像上测量增强扫描的预计扫描范围,即气管隆突下1 cm至心脏下缘2 cm.比较两种扫描范围及相应的辐射剂量的差异.结果:实际扫描范围比预计扫描范围平均缩短(2.09±0.76) cm(t=17.31,P＜0.01),冠状动脉CTA各序列总剂量较常规方法平均减少(1.51±0.87)mSv,差异有极显著性意义(t=10.92,P＜0.01),相当于各序列平均剂量总和的9.20％.结论:使用钙化积分扫描图像能有效缩短增强扫描的范围,从而明显降低冠状动脉CT血管成像的辐射量.     

钙化积分扫描缩短冠脉CTA扫描范围及降低辐射量的作用

目的:探讨使用钙化积分扫描缩短增强扫描范围以降低后门控冠状动脉CTA检查辐射量的临床应用价值。方法:40例患者行冠脉钙化积分及冠脉CTA检查,其增强扫描的实际扫描范围根据钙化积分扫描图像来决定,即冠状动脉树上缘1cm至心尖部心包影即将消失的层面。并按常规方法在定位像上测量增强扫描的预计扫描范围,即气管隆突下1cm至心脏下缘2cm。比较两种扫描范围及相应的辐射剂量的差异。结果:实际扫描范围比预计扫描范围平均缩短(2.09±0.76)cm(t=17.31,P<0.01),冠状动脉CTA各序列总剂量较常规方法平均减少(1.51±0.87)mSv,差异有极显著性意义(t=10.92,P<0.01),相当于各序列平均剂量总和的9.20%。结论:使用钙化积分扫描图像能有效缩短增强扫描的范围,从而明显降低冠状动脉CT血管成像的辐射量。     

尘肺CT扫描方案的探讨

目的比较常规剂量CT、低剂量CT及高分辨率CT（HRCT）3种扫描方案对尘肺的诊断能力,以期确定最佳方案。方法 37例经专业机构确诊的尘肺患者均经常规剂量CT、低剂量CT及HRCT3种方案扫描。采用统计学方法分析与比较该3种方案对尘肺特征性表现,小阴影、大阴影及进行性团块状纤维化（PMF）及纤维化早期表现及合并症的检出率。结果 3种扫描方案对小阴影、大阴影及团块状纤维化（PMF）的检出例数相符,均为36例、20例及16例,而对于大、小阴影及PMF周围的瘢痕肺气肿、肺纹理扭曲等征象,以HRCT显示较清晰。HRCT对早期肺间质纤维化,诸如胸膜改变、小叶间隔增厚、小叶肺气肿及蜂窝状影的显示例数显著多于其他两组（P〈0.05）。除肺气肿外,3种方案对其他合并症肺结核、肺癌及呼吸系统感染等的检出率无明显差别。结论低剂量CT与常规剂量CT均可用于尘肺的普查与诊断,为减少受检者的辐射剂量,应尽可能采用低剂量CT扫描。而对于肺间质改变的观察,HRCT应作为首选方案。