X射线致HeLa细胞损伤过程中NADPH氧化酶的作用

以HeLa细胞为实验材料, 探讨了NADPH氧化酶在X射线诱导细胞损伤过程中的作用。 结果显示, 12 Gy X射线辐照后细胞内活性氧(ROS)明显增加, 在用NADPH氧化酶抑制剂处理后再辐照, 则细胞内ROS降低到未辐照水平; 同时辐照后NADPH 氧化酶细胞质亚基p47phox 在细胞质积聚并和细胞膜亚基 gp91phox 结合; Western blotting检测结果显示, NADPH 氧化酶的关键亚基 gp91phox 的表达量明显增加。 以上结果说明, NADPH氧化酶可以被X射线激活, 由其介导产生的ROS在X射线诱导HeLa细胞损伤过程中扮演重要角色。 To investigate the role of NADPH oxidase in HeLa cell lesion induced by X ray irradiation, the change of cell survival was detected with MTT assay, reactive oxygen species (ROS) was measured by fluorospectrophotometer. Immunostaining and confocal laser scanning microscopy was employed to detect the co localization of two subunit of NADPH oxidase, p47phox and gp91phox in the cell. Western blotting was used to detect the expression of gp91phox before and after X ray irradiation. After X ray irradiation, intra cellular level of ROS increased obviously. But the increase could be blocked by diphenyleneiodonium (DPI), an inhibitor of NADPH oxidase. Meanwhile, cytosolic subunit p47phox moved to membrane and co localizated with gp91phox after irradiation. Moreover, the results also show that gp91phox increased sharply after 12 Gy X ray irradiation. Therefore, NADPH oxidase mediated production of ROS plays an important role in HeLa cell lesion induced by X ray.     

羧甲基-β-1,3-葡聚糖对人肝癌HepG2细胞的放射增敏作用

本研究旨在探讨羧甲基-β-1,3葡聚糖（CMG）对人肝癌HepG2细胞X射线或12C6+离子束辐射敏感性的影响。首先用CCK-8法检测CMG对HepG2细胞的生长抑制情况,得到半数抑制浓度（IC50）为120.6μg/mL。用浓度为0.1×IC50的CMG预处理HepG2细胞24 h,再给予2 Gy X射线或12C6+离子束辐照（CMG+辐照组）;CMG未处理组直接接受2 Gy X射线或12C6+离子束辐照（辐照组）。对比分析辐照组和CMG+辐照组细胞的克隆存活、DNA损伤、凋亡与周期分布、细胞内活性氧（ROS）水平。发现:与X射线辐照组相比,相同剂量的12C6+离子辐照组克隆存活率更小,DNA损伤和周期阻滞更加严重,细胞凋亡率和细胞内ROS水平也更高。与单独X射线或12C6+离子束辐照组相比,CMG+辐照组克隆存活率明显降低,细胞凋亡率随辐照后CMG作用时间的延长而明显增加,CMG使辐照后细胞内ROS维持在一个较高的水平,同时CMG明显加重了单独辐照诱导的DNA损伤和周期阻滞。结果表明,与X射线相比,HepG2细胞对相同剂量的12C6+离子辐射更敏感;CMG可增加HepG2细胞对X射线或12C6+离子辐射的敏感性;CMG可能通过增加受照HepG2细胞内的ROS水平,加剧辐照诱导的DNA损伤,促进辐射诱导细胞凋亡而起到辐射增敏作用。This study aims to investigate the effect of carboxymethy-β-1, 3-glucan (CMG) on the sensitivity of human hepatoma HepG2 cells to X-rays or 12C6+ ions irradiation. First, the inhibitory effect of CMG on the growth of HepG2 cells was detected by CCK-8 assay, and the half maximal inhibitory concentration (IC50) was 120.6 μg/mL. HepG2 cells were pretreated with CMG at a concentration of 0.1×IC50 for 24 h and then irradiated with 2 Gy X-ray or 12C6+ ion beams (CMG + irradiation group). CMG untreated group was directly irradiated by 2 Gy X-rays or 12C6+ ions beam (irradiation group). The clone survival, DNA damage, cell apoptosis, cell cycle distribution, and intracellular reactive oxygen species (ROS) levels in irradiation group and CMG + irradiation group were comparatively analyzed. The results showed that the clone survival rate was lower, DNA damage and cycle arrest were more serious, and the rate of apoptosis and intracellular ROS levels were higher in 12C6+ ions irradiation group than those in the same dose of X-rays irradiation group. Compared with X-rays or 12C6+ ions irradiation group, the clone survival rate of CMG + irradiation group was significantly decreased, and the apoptosis rate significantly increased with the prolongation of CMG treatment post-irradiation; CMG maintained intracellular ROS at a higher level after irradiation, CMG also significantly aggravated radiation-induced DNA damage and cycle arrest. These results indicated that HepG2 cells were more sensitive to 12C6+ ions radiation than those at the same dose of X-rays. CMG increased the sensitivity of HepG2 cells to X-rays or 12C6+ ions irradiation by increasing intracellular ROS level, exacerbating radiation-induced DNA damage and promoting radiation-induced apoptosis in irradiated HepG2 cells.     

X射线诱导的HeLa细胞旁效应中ROS和NO关系研究

活性氧(ROS)和一氧化氮（NO)是辐射诱导的旁效应信号通路中的两个重要信号分子。 实验研究了这两种信号分子在HeLa细胞旁效应信号通路中的关系。 通过微核实验, 发现X射线辐照过的HeLa细胞及其旁观者细胞微核形成明显增加, 而二甲亚砜（DMSO）预处理显著抑制了微核形成。 另外还发现, 接受条件培养基的旁观者细胞的增殖速率增加, 而DMSO预处理产生条件培养基的受辐照细胞则使旁观者细胞的增殖速率降低。 以上的结果从不同角度证实了HeLa细胞存在X射线诱导的旁效应, 且其可以被DMSO预处理所抑制。Western blotting和DAF FM DA荧光探针检测分别显示出辐照后细胞的诱导型一氧化氮合酶（iNOS）和NO水平均升高, 而DMSO预处理则降低其水平。 因此, 可以推测X射线诱导的HeLa旁效应当中ROS是NO的上游信号。 Accumulating evidence indicates that irradiated cells can release signals which induce a series of biological responses in non exposed cells. This is known as irradiation induced bystander effects. Both reactive oxygen species(ROS) and nitric oxide(NO) play important roles in bystander effects. In this study, we determined the relationship of ROS and NO in the signaling pathway of bystander effects. HeLa cells were treated with or without dimethye sulfoxide(DMSO) before X ray irradiation, and micronuclei formation as well as cell proliferation rate was detected in both irradiated and bystander cells. In addition, we also detected inducible nitric oxide synthase(iNOS) expression and NO level in irradiated cells using Western blotting and DAF FM DA fluorescent probe, respectively. Our results showed that micronuclei were induced in irradiated and bystander cells while DMSO treatment significantly suppressed the formation of micronuclei in both of them. We also found that when cells were irradiated their proliferation rate was suppressed while DMSO treatment eliminated this inhibition effect. In contrast, the cells received conditioned medium from irradiated cells proliferated more quickly than the cells received medium from non irradiated cells while DMSO treatment reduced the difference. Finally, we found that irradiated cells had higher level of iNOS and NO compared to non irradiated controls, whereas DMSO treatment decreased their levels. These results suggest that ROS is the upstream signal of NO in X ray induced bystander effects in HeLa cells.     

重离子诱导细胞衰老研究

综述了细胞衰老诱导因素和衰老细胞的特征,提出细胞衰老在癌症发生发展过程中具有双重作用效应。同时,以X射线和12C6+离子束对黑色素瘤细胞系92-1或人正常成纤维细胞系MRC5进行辐照,观察分析DNA损伤应答效应和细胞衰老诱导。实验结果表明,相对于X射线,12C6+离子束能导致DNA团簇损伤,持续激活DNA损伤应答,更容易诱导细胞衰老;12C6+离子束能同时诱导癌细胞和正常细胞衰老。这些实验结果暗示开展12C6+离子束诱导细胞衰老研究具有紧迫性。     

X射线不同方向辐照80C196KC20单片机研究

 为明确X射线不同方向入射对器件的影响,利用中国测试技术研究院的直流SJ-A X光机产生的X射线,从与垂直80C196KC20单片机表面不同方向角度对单片机进行辐照试验。试验结果显示,随着辐照角度由0°增加到90°,单片机所受的影响逐渐减小,90°时最不明显,表明此种单片机外壳在对其X射线总剂量效应的影响最大,原因在于X射线在不同方向照射时,需要穿过不同厚度的封套材料,X射线在此过程受到的衰减不同。     

γ射线辐照处理竹材的X射线光谱研究

利用X射线衍射仪,对γ射线辐照处理前后的竹材进行X射线光谱分析,得出竹材结晶度和微纤丝角在辐照过程中的变化规律。随着辐照剂量的增加,竹材纤维素结晶度呈现先升高后降低的趋势,微纤丝角变化不明显,表明微纤丝角不是影响辐照过程中竹材物理、力学性能变化的主要因子。     

氧化钆纳米粒子对A549肺癌细胞的辐射增敏效应研究

采用氧化钆纳米粒子（GON）,研究钆基纳米粒子对X射线和碳离子束的辐射增敏效应。首先,通过透射电镜观察材料粒径,使用DLS检测材料的水合半径及Zeta电位,并用紫外吸收谱证实GON在培养基中稳定性较好;研究发现钆（Gd）浓度为10.0 μg/mL的GON对30 keV/μm碳离子束辐照水溶液产生的羟自由基的增强系数为1.13;GON对A549肺癌细胞和正常MRC-5肺细胞没有明显的毒性,且在人肺癌A549细胞中的摄取量随共培养浓度的增加而增加,在10.0 μg/mL共培养浓度下,细胞摄入Gd的量为0.73 pg/cell;进一步采用克隆存活实验证明,GON的加入对X射线和碳离子辐照A549细胞所产生的损伤具有明显的增强,在10%的细胞存活水平下,GON对A549细胞在X射线及碳离子辐照下的辐射增敏分别达15.5%和10.1%。鉴于钆材料常被用于磁共振成像（MRI）,所获得的GON有望作为X射线和碳离子的诊疗一体化材料。     

12C和X射线辐照人肺癌细胞H1299的生物学效应

用高传能线密度（LET）的12C离子束和低LET的X射线辐照体外培养的非小细胞肺癌H1299(p53基因缺失), 研究它们的辐照生物学效应的差异。 用克隆形成率法测定了细胞对射线的辐射敏感性; 用AnnexinV/PI试剂盒检测了细胞早期凋亡; 用流式细胞仪检测了细胞周期变化。 实验结果表明, 12C离子束辐照H1299细胞的存活率明显低于用X射线辐照的; 12C离子束引起H1299细胞的早期凋亡率明显高于X射线辐照引起的, 且持续时间更长; 12C离子束引起的H1299细胞G2/M期的抑制更明显。 说明H1299细胞对高LET的12C离子束的辐射敏感性高于对X射线的, 重离子对p53基因缺失型肿瘤的治疗可实施较低的照射剂量、 较少的照射次数和较长的时间间隔。     

低强度激光辐照HeLa细胞诱导增殖效应的可见光谱特性

比较研究了可见波段405,514,633和785nm低强度激光辐照HeLa细胞的促增殖效应。实验采用100和1 000J.m-2两种有效能量密度的可见光波辐照细胞,照射后24,48,72h采用MTT法检测细胞活性。结果表明,405,633,785nm激光均能促进HeLa细胞增殖,且呈波长和时间依赖性;633nm激光辐照对细胞促增殖效应最显著;514nm激光辐照对HeLa细胞促增殖效应较不明显。不同光剂量对细胞增殖效应不同,405,633,785nm激光辐照时能量密度为1 000J.m-2的辐照组细胞增殖均较100J.m-2辐照组明显,而514nm激光辐照时1 000J.m-2辐照组与100J.m-2辐照组细胞增殖的差异不明显。     

电缆X射线辐射感应电导率引起的非线性效应北大核心CSCD

对电缆X射线辐照非线性响应进行了数值模拟。分析了电缆X射线辐照的物理过程,针对同轴电缆和屏蔽多导体电缆,建立了基于有限元方法的二维电缆模型诺顿等效电流源计算方法,着重描述了二维模型下电缆介质辐射感应电导率效应求解方法,模拟了电缆X射线辐照非线性效应,给出了诺顿等效电流源非线性响应规律,并依据物理过程进行了合理性分析。模拟结果显示,由于辐射感应电导率的存在,随着X射线注量的增加,电缆响应幅度会存在明显的饱和现象。对特定类型的电缆,响应电流随着X射线注量增加,依次出现3个时间位置不同的电流峰值。     

内质网应激对碳离子辐照宫颈癌HeLa 细胞的影响

利用不同剂量的碳离子辐照二硫苏糖醇(2.5 mmol/L) 预处理的HeLa 细胞,探讨了内质网应激反应对碳离子辐照宫颈癌HeLa 细胞的影响。实验发现:与单独辐照组相比,二硫苏糖醇联合碳离子辐照后细胞的存活率下降,而凋亡率增加;二硫苏糖醇联合碳离子辐照加重了碳离子辐照引起的细胞周期阻滞;且联合辐照组的自噬被明显激活。结果表明,持续的内质网应激可改变宫颈癌HeLa 细胞对碳离子辐照反应,且二硫苏糖醇可能通过影响HeLa 细胞的自噬性细胞死亡通路发挥作用。To investigate the effect of endoplasmic reticulum stress on HeLa cells to 12C6+ ion irradiation,HeLa cells were pretreated with 2.5 mmol/L dithiothreitol and irradiated by 12C6+ ions with different doses.The results showed that, compared with IR alone, dithiothreitol combined with carbon ion irradiation caused HeLa cell survival decreased, and the apoptosis increased. Moreover, dithiothreitol and carbon ion radiation combination treatment led to a significant increase of G2/M phase, and autophagy was activated obviously in combination treatment group. The results imply that continuous endoplasmic reticulum stress can change the response of HeLa cells to 12C6+ irradiation, and dithiothreitol may affect HeLa cells through the autophagy cell death pathway.     

^12C和X射线辐照人肺癌细胞H1299的生物学效应

用高传能线密度（LET）的^12C离子束和低LET的X射线辐照体外培养的非小细胞肺癌H1299（p53基因缺失）,研究它们的辐照生物学效应的差异。用克隆形成率法测定了细胞对射线的辐射敏感性;用AnnexinV/PI试剂盒检测了细胞早期凋亡;用流式细胞仪检测了细胞周期变化。实验结果表明,^12C离子束辐照H1299细胞的存活率明显低于用X射线辐照的;^12C离子束引起H1299细胞的早期凋亡率明显高于X射线辐照引起的,且持续时间更长;^12C离子束引起的H1299细胞G2/M期的抑制更明显。说明H1299细胞对高LET的^12C离子束的辐射敏感性高于对X射线的,重离子对p53基因缺失型肿瘤的治疗可实施较低的照射剂量、较少的照射次数和较长的时间间隔。     

BaCl2微晶辐照损伤的电子自旋共振研究

用电子自旋共振(ESR)方法研究发光基质材料BaCl₂的X射线和γ射线辐照损伤及其恢复的情况,发现样品由X射线引起的辐照损伤,在日光下仅两分钟便能完全恢复;而γ射线引起的辐照损伤则很难完全恢复。 关键词：     

真空环境中紫外脉冲激光对熔石英抗损伤能力的影响

在10-3Pa的高真空环境下,利用355nm的紫外脉冲激光以低于石英基片零概率损伤阈值的能量密度对其进行重复多次的全口径扫描,目的是为了研究石英基片在接受不同剂量的紫外激光辐照后其抗损伤能力的变化和原因.通过辐照过程中损伤点的扩展程度以及辐照后的抗损伤测量表明,高真空环境下紫外脉冲激光辐照后的石英基片抗损伤能力明显降低,降低的幅度更多的与其接受激光辐照的次数有关,辐照能量密度的影响相对较小;通过荧光及X射线光电子能谱(XPS)检测发现,紫外脉冲激光辐照后石英基片表面氧缺位的增加是导致其抗损伤能力下降的主要原因,其程度也直接与接受辐照的紫外脉冲激光剂量相关.     

低剂量电离辐射预处理对不同肿瘤细胞周期的影响

实验研究了低剂量γ射线预照射对人肝癌细胞系hepG2和人宫颈癌细胞系HeLa的细胞周期进程的影响.结果显示(1)低剂量(5cGy)辐射后,两种细胞的G2/M期细胞短暂累积;(2)低剂量辐射促进肿瘤细胞的生长;(3)高剂量(3Gy)辐射后,hepG2细胞发生G2期阻滞,HeLa细胞发生S期和G2期阻滞;(4)与单纯高剂量照射相比,低剂量辐射预处理后4h,再给予高剂量辐射,可进一步促进hepG2细胞在G2/M期累积,但是预照射对HeLa细胞的周期进程没有明显影响.因此,低剂量辐射预处理对高剂量诱导的细胞周期阻滞的影响依赖于肿瘤细胞的类型.     

强脉冲X射线辐照Si-SiO2界面对C-V 和I-V特性曲线的影响

 利用强脉冲X射线对Si-SiO₂界面进行了辐照，测量了C-V曲线和I-V曲线。实验发现，经过强脉冲X射线对Si-SiO₂界面进行的辐照，使C-V曲线产生了正向漂移，这一点与低剂量率辐射结果不同；辐射后，感生I-V曲线产生畸变；特别地，从I-V曲线上还反映出强脉冲X射线辐照的总剂量效应造成电特性 参数明显退化，最后甚至失效。讨论了强脉冲X射线辐照对Si-SiO₂界面产生损伤的机理，并对实验结果进行了解释。     

GANRA-5对X射线和~(12)C~(6+)离子辐射防护作用研究

研究了一种新的恶唑酮类衍生物GANRA-5对于人胚肺细胞MRC-5的辐射防护作用。以MTT评价其对于细胞的毒性,以γH2AX foci形成法检测其对于辐照后细胞中双链断裂的影响,发现其对于受到X射线和12C6+离子照射的细胞具有较强的辐射防护作用,并进一步发现其能够显著清除辐照后细胞内的自由基。这些结果表明,GANRA-5具有较低的细胞毒性,并能够通过清除自由基发挥较强的针对X射线和12C6+离子的辐射防护作用,有望开发为高效的辐射防护药物。     

辐照诱导的人正常肝细胞系HL-7702细胞延迟效应

利用X射线辐照人正常肝细胞系HL-7702细胞, 运用胞质分离阻滞微核法实验检测细胞微核率,AnnexinV FITC细胞凋亡检测试剂盒检测细胞凋亡率, 细胞微核率和凋亡率随着辐照剂量的增加而显著增加。X射线照射后细胞传代培养, 第7代时不同剂量辐照后子代细胞微核率和凋亡率同未辐照细胞相比已无明显区别。 对不同剂量辐照后传代7代的细胞再次照射2.5 Gy的相同剂量,发现它们细胞微核率和凋亡率存在明显差异,即初次受辐照剂量高的细胞, 再次以相同剂量辐照后的微核率和凋亡率也高. 这些结果表明,X射线辐照导致了HL-7702细胞基因组不稳定性这一辐射延迟效应,再次辐照使得辐射的延迟效应得以明显的表现。 Human normal liver cell line HL-7702 cells were irradiated with different doses of X rays. Micronucleus and apoptosis rates in the irradiated cells were measured with cytokinesis block micronucleus method and Annexin V FITC apoptosis detection kit, respectively. Experimental data showed that the micronucleus and apoptosis rates increased obviously with increasing irradiation dose. After seven population doublings, the micronucleus and apoptosis rates of the cells surviving exposure to the X rays reduced to the same levels as non irradiated control cells; the progenies of the cells were secondly exposed to X rays at the same dose of 2.5 Gy. We found that the progenies of the cells surviving the first irradiations of the various doses showed markedly differential micronucleus frequencies and apoptotic rates. Although the same dose of 2.5 Gy was applied in the second irradiations, the micronucleus frequencies and apoptotic rates of the progenies of the cells initially exposed at higher doses were significantly higher than the others. These results indicate that X rays lead to genomic instability in HL 7702 cells, which is an important manifestation of radiation induced delayed effect, and a second radiation stimulus makes the delayed effect in the progeny of the previously irradiated cells be expressed obviously.     

X射线辐照对发根农杆菌介导芸芥遗传转化的影响

探讨了X射线辐照对芸芥发状根诱导的影响, 为研究辐射对转基因技术的协同作用提供一定的实验依据。 以芸芥无菌苗的子叶为材料, 通过5—20 Gy的X射线辐照和不等浸染时间的联合处理, 研究苗龄、 预培养时间、 辐照剂量及其浸染时间等因素对芸芥发状根诱导的影响作用, 并通过聚合链酶式反应（PCR）对所得发状根进行了分子水平的鉴定。 15 Gy X射线辐射能提高芸芥发状根的诱导频率, 且有量效关系, 其中先浸染后辐照比先辐照后浸染效果更显著; PCR结果也表明, 发根农杆菌R1000的 rolB基因已成功地被转入并整合到该发状根的基因组中。 X射线辐照对芸芥发状根的诱导具有一定的促进作用, 且最佳推荐诱导剂量为15 Gy或稍多。     

带状电缆的直流X射线辐照响应

对真空环境下带状电缆模型直流X射线辐照响应进行了实验和数值模拟研究;研制了电缆直流X射线辐照实验系统;使用蒙特卡罗模拟软件计算了直流X光机产生的X射线能谱、通量等参数;建立了带状电缆X射线辐照响应一维计算模型,该计算模型包括电缆导体与介质层间隙和介质层电导率模型。实验测量了两个带状电缆模型的直流X射线辐照响应电流波形,并对其进行了数值模拟。结果显示,在一定的电缆导体与介质层间隙大小假设条件下,采用带状电缆X射线辐照响应计算模型计算的结果与实验测量结果在波形特征和绝对幅度方面比较接近,说明了利用该模型描述电缆直流X射线辐照响应具有其合理性。