^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。     

不同LET射线对细胞的生物学效应

以人肝癌细胞系和正常肝细胞系为材料,报道了不同传能线密度射线辐射引发细胞染色体原初断裂及24 h内的修复情况。 计算了相对生物学效应的值。 以L02染色体总断裂数量得出的RBE值96.05 keV/μm的12C6+ 为3.6, 512 keV/μm 36Ar18+ 为2.9。 而以7721染色体总断裂数量得出的RBE值: 96.05 keV/μm的12C6+ 为3.5,512keV/μm 36Ar18+也为2.9。用产生等点染色单体断裂计算,则RBE更高。对比得出,高LET对增加等点染色单体断裂量的作用要远远大于对增加染色单体断裂量的作用。等点染色单体的断裂修复难度要远远大于染色单体断裂的修复难度, 这也是高LET高致死率的一个重要原因。 Human hepatoma SMMC 7721 and normal liver L02 cells were irradiated with γ rays,12C6+ and 36Ar18+ ion beams at the Heavy Ion Research Facility in Lanzhou(HIRFL). We reported the kinetic repair of chromosome breaks of L02 and SMMC 7721 cells in 24 h of post irradiation time. The relative biological effectiveness(RBE) for inducing chromatid breaks were 3.6 for L02 and 3.5 for SMMC 7721 cell lines at the linear energy transfer(LET) peak of 96.55 keV/μm 12C6+ ions, and 2.9 (both of the two cell lines) at 512 keV/μm 36Ar18+ ions.It suggested that the RBE of isochromatid type breaks induced by 36Ar18+ was higher than those by 12C6+. We concluded that the high production of isochromatid type breaks, induced by the densely ionizing track structure, could be regarded as a signature of high LET radiation exposure.     

碳离子辐照细胞的相对生物学效率

研究了传能线密度(LET)≥125.5keV/μm的碳离子辐照小鼠黑色素瘤B16、人的宫颈癌HeLa、中国仓鼠肺V79、人的肝癌SMMC-77214种细胞的相对生物学效率(RBE).得到了当LET=125.5keV/μm时,RBE依赖于细胞种类并随细胞存活水平的升高而增加的关系,以及当LET≥125.5keV/μm时,RBE随着LET的增大而变小的关系.     

不同LET 12C6+离子对生防菌BJ1的辐射诱变效应

选取高低两个LET点(40和60keV/m),剂量点分别为50,100,200,400,600Gy进行辐照处理,研究了生防菌的存活率与突变率的关系,抑菌谱以及活性等。结果表明,在高LET条件下,低剂量辐照就可以得到较多的突变体,并且BJ1有较高的存活率和突变谱,有利于筛选优良的正突变体。因此高LET较低LET有更为明显的辐射诱变效应。     

辐射诱导的DNA修复与细胞凋亡的ATP调控

综述了DNA辐射损伤导致的细胞阻遏于G1期以及在该时期对DNA的修复活动, 提出了较大剂量辐射诱导的三磷酸腺苷不足导致细胞凋亡的假说, 并分析了细胞走向凋亡与修复的辨正关系。 DNA damage induced by irradiation,which makes the cell arrested at G1 stage and DNA repair being activated in this stage,are summarized. It is proposed that the deficiency of adenosine triphosphate which is induced by the larger irradiation dose, induces cell apoptosis. And the relationship of cell selecting repair and apoptosis is also analyzed.     

不同LET C离子束对粘红酵母菌的突变效应分析

以粘红酵母菌Rhodotorula glutinis AY 91015为材料, 研究了不同传能线密度（LET）的C离子对粘红酵母菌的失活截面和突变截面, 评估了不同LET的C离子对微生物的失活效应和突变效应。 结果表明, C离子LET为120.0 keV/μm时, 单个粒子对粘红酵母菌的失活截面最大, 为4.37 μm2, 接近酵母菌细胞核的平均核截面; LET为96.0 keV/μm时, 单个粒子对粘红酵母菌的突变截面最大。 通过对C离子束致突变能力的分析发现, C离子在LET为58.2 keV/μm时突变能力最强, 这一结果显示在经C离子辐照后存活下来的粘红酵母菌中, 可以引起有效突变的最佳LET为58.2 keV/μm左右, 此时所对应的碳离子能量约为35 MeV/u。 这些结果表明, C离子对粘红酵母菌的最佳致死效应和最佳致突变效应存在于不同的能量区域。 To evaluate inactive and mutagenic effects of carbon beam at different LET, the inactivation cross section and mutation cross section induced by carbon beams of different LET values were investigated in a red yeast strain Rhodotorula glutinis AY 91015. It was found that the maximum inactivation cross section of 4.37μm2 , which was very close to the average nucleus cross section, was at LET of 120.0 keV/μm. The maximum mutation cross section was at LET of 96.0 keV/μm. Meanwhile, the highest mutagenicity of carbon ion was found around 58.2 keV/μm. It implied that the most efficient LET to induce mutation in survival yeasts was 58.2 keV/μm, which corresponded to energy of 35 MeV/u carbon beam. The most effective carbon beam to induce inactivation and mutation located at different energy region.     

碳离子辐照对人肺癌细胞A549细胞周期进程的影响

选取对数生长期人肺癌细胞A549接受0—6.0 Gy 碳离子照射， 用克隆形成法检测细胞的存活率； 并于照射后12和24 h收集细胞， 用流式细胞术检测细胞周期各时相的细胞百分比， 观察不同剂量碳离子辐照对A549细胞周期进程的影响。 结果显示: 0—6.0 Gy 碳离子照射后细胞存活率显著下降； 照射后12 h细胞发生G0/G1期阻滞， 而照射后24 h， 1.0 Gy 照射组细胞在G0/G1期阻滞， 2.0—6.0 Gy 照射组细胞在G2/M期阻滞。 上述结果表明， 在A549细胞接受碳离子照射后的12 和24 h内， 1.0 Gy 照射可持续激活细胞G1期检查点， 而2.0—6.0 Gy 碳离子照射后其细胞周期进程是随时间变化的。 To investigate the effects of cell cycle progression of A549 cell induced by 12C6+ ion irradiation at different doses, the survival fractions of the A549 cells were determined by colony forming assay; cell cycles were analyzed by FACS at 12 h or 24 h after irradiation. The results showed that the percentage of survival in the A549 cells decreased with irradiation doses. Compared with control group， the percentage of the cells in G0/G1 phase significantly increased at 12 h after irradiation with different doses of 12C6+ ions. However， at 24 h after irradiation the percentage of the cells in G0/G1 phase significantly increased with 1.0 Gy 12C6+ ions， while the cells showed increasing percentage in G2/M phase with 2.0, 4.0 or 6.0 Gy 12C6+ ions. The results suggested that G1 cell cycle checkpoint was activated in 12—24 h after irradiation with 1.0 Gy 12C6+ ions， but after irradiation with 2.0—6.0 Gy 12C6+ ions， the cell cycle progression of the A549 cells changed with time.     

7Li和12C离子致DNA链断裂的研究

选用HI-13串列加速器产生的不同传能线密度的⁷Li和¹²C重离子,以不同的剂量对纯化的质粒DNA水溶液进行了辐照.利用原子力显微镜对这两种重离子诱发的DNA损伤进行了纳米水平的结构分析,并用ScionImage分析软件完成了DNA碎片长度的测量.得到了DNA分子超螺旋、开环和线性三种形态随剂量的变化情况以及DNA碎片长度的分布函数,并用Tsallis熵统计理论对实验结果进行了拟合.     

人肝癌SMMC-7721细胞对低剂量γ射线辐射超敏感性的研究

研究了人肝癌细胞SMMC-7721在低剂量γ射线照射下超敏感性和增强的辐射抗性响应。选用对数生长期细胞接受0—6 Gy不同剂量的^60Coγ射线的照射。利用流式细胞仪对细胞进行分选计数,并用克隆形成法检测细胞存活率。发现SMMC-7721细胞存在低剂量辐射超敏感性和增强的辐射抗性响应,即在0—0．3Gy之间细胞表现出单位剂量杀伤增强现象,在0．3—1Gy细胞表现一定的辐射抗性,在1Gy以上,细胞的存活符合线性平方模型。     

6MeV/u 12C6+离子对四种作物种子的生物学效应研究

在中科院近代物理研究所兰州重离子研究装置(HIRFL)上,采用46.6MeV/u~(12)C~(6+)离子对四种不同作物种子进行了生物学效应研究,结果表明:重离子的轰击和贯穿,不仅会在种子表面外貌而且也对种子内部细胞造成严重损伤,重离子对种子萌发能力和生长发育均有明显的抑制作用,对生物功能也有影响;在种子的根尖细胞中出现的染色体畸变具有多种类型,而且畸变频率明显高于对照组。重离子对多种细胞器也产生了严重损伤,受损DNA具有一定的修复能力,测定了四种作物种子的半致死剂量及失活截面,找到了四种作物之间辐射敏感性的差异,讨论了细胞器损伤与生理过程异常的关系和受损DNA的修复过程。 The biological effects of 46.6MeV/u ~(12)C~(6+) ions on four kinds of plant seeds were studiedat HIRFL of Institute of Modern Physics (IMP), Academia Sinica. The results indicate that seriousinjuries were presented on not only external features of the seeds but also internal cells due to the bombardment and penetration of the ions. The heavy ions can significantly inhibitgermination and growth of the seeds and can affect physiological functions. In root tip cells ofirradiated seeds...     

彗星电泳检测 12C6+离子束辐照人类肝L02细胞的DNA损伤效应

以传能线密度为30 keV/μm的12C6+离子束辐照人类肝L02细胞, 利用彗星电泳技术检测了以DNA链断裂为生物终点的DNA辐射损伤效应。 CASP软件分析彗星图像, 主要检测尾部DNA（TDNA％）、 彗星全长（CL）、 尾长（TL）、 尾矩（TM）和Olive尾矩（OTM）等指标, SPSS 11.5软件进行统计学分析, 绘制并拟合TM\|剂量曲线。 结果显示, 辐照以剂量依赖的方式引起L02细胞彗星图像各指标的增大, 且TM值与剂量线性正相关。 说明12C6+离子束对DNA有较强的致损伤效应, 且与剂量正相关。 研究为正确评价重离子对人体正常组织的辐射风险及危害提供了一定的基础数据和依据。     

12C离子束的剂量学研究

研究了L-α-丙氨酸剂量计测量^12C离子辐射的剂量学特性，实验证明丙氨酸剂量计适用于^12C离子辐射的剂量学测量。另外，还研究了^12C离子照射人外周血诱发的染色体畸变（双着丝粒＋着丝粒环）的剂量效应，在0-8．0Gy范围内拟合的最佳回归方程为Y=0．858503D＋0．3615×10^-2D^2。Dosimetric characteristics of L-α-alanine dosemeter used for dosimetry of ^12C ion radiation have been studied. The experimental results indicate that the alanine dosemeter can be used to measure the ^12C ion radiation. In addition, dose effects of chromosome aberration dicentrics and cenric rings were studied after human peripheral blood being irradiated by ^12C ions; the best regression equation, Y = 0. 858 503D ＋ 0. 361 5 ×10^-2D^2, was obtained within 8.0 Gy.     

放射性束辐照生物学效应及相关生物物理机理

介绍了β缓发粒子衰变放射性束 9C辐照处于不同贯穿深度上人类唾液腺细胞的深度存活效应。 与稳定的 12C束流相比, 放射性 9C束流在其Bragg峰区附近展示了增强的生物学效应, 即细胞的致死效率明显增强。 探讨了放射性束 9C在其Bragg峰区展示增强生物学效应的生物物理机制, 并介绍了利用另一种放射性束 8B进行辐照生物学效应研究的新进展。 最后, 提出了一些利用放射性束可进行的辐照生物学效应前沿研究。 The depth survival effect of a β delayed particle decay 9C beam on human salivary gland (HSG) cells is presented in this paper. Compared with a stable 12C ion beam, the radioactive 9C ion beam showed an enhanced biological effect, i.e., remarkably enhanced efficiency in cell killing, at the penetration depths around its Bragg peak. The biophysical mechanisms underlying the enhanced biological effect are discussed herein, and the latest progress in biological effect induced by another radioactive 8B ion beam is introduced. In the end of this paper, several topics concerning the frontier of the radiation induced biological effects using radioactive ion beams are proposed.     

大分割碳离子放射治疗中相对生物学效应与剂量依赖关系的研究

应用碳离子束进行大分割放射治疗从而缩短治疗周期是碳离子束治疗的优势之一。为研究大分割放疗增加单次照射剂量后,碳离子束相对生物学效应（RBE）的变化,应用细胞存活线性平方（LQ）模型推导出RBE与剂量的依赖关系。基于此关系研究了具有不同辐射敏感性的肿瘤细胞和正常组织细胞RBE随剂量的变化。结果表明,在0~20 Gy范围内,不论肿瘤细胞与正常组织细胞具有怎样的辐射敏感性,肿瘤细胞的RBE值始终大于正常组织细胞。此外,基于理论推导和对相关实验数据的分析,证实了RBE随剂量增加而递增现象的存在。这些结果对应用碳离子束进行大分割放射治疗具有重要的指导作用。Short treatment course due to the suitability of hypofractionated regimen for carbon ion beam is one of the advantages of carbon ion radiotherapy. To study the dependence of relative biological effectiveness (RBE) on dose, the relationship between RBE and dose was deduced through the linear-quadratic (LQ) model. Based on the relationship, the change of RBE of tumor cells and normal tissue cells with different radiosensitivities with dose was studied. The results showed that the RBE value of tumor cells was always greater than that of normal tissue cells in the dose range of 0~20 Gy, regardless of the radiosensitivity of tumor and normal tissue cells. In addition, based on theoretical deduction and analysis of the relevant experimental data, the existence of RBE increase with increasing dose was verified. These results are of great significance for conducting hypofractionated radiotherapy with carbon ion beam.     

离子束介导技术在生物体遗传改良上的发展趋势

在利用离子束介导技术对生物体进行遗传改良的研究领域内仍然存在着7大研究难题值得注意。 明确提出了离子束介导技术进一步发展的技术思路, 即立足于离子束介导技术这一物理学技术平台, 注重研究两个关键性问题(即离子束介导异源遗传物质进入受体基因组的机理和受体发生异源基因重组的机制)。 在研究中完成3个有效转变(从形态学鉴定为主有效地转变为对其遗传学规律进行研究, 从对介导当代群体的变异效应的研究为主有效地转变为研究突变性状在多个世代内所表现的后效性, 从对单一性状的研究为主有效地转变为研究突变性状的综合表现), 寻找4个方面的实验证据(形态学、 生理生化、 细胞和分子生物学的证据), 研究5大生物学特性(生殖特性、 发育特性、 光合特性、 抗逆性和品质等特性)。 The 7 research puzzles in the genetic improvement of biological bodies made by ion beam mediated technique, are worth noticed. The technical ideas, including one mediated technique in physics, 2 significant subjects, 3 effective changes, the mediated evidences of 4 aspects and 5 biological characteristics, were particularly put forward according to the existing states in the field. The 2 significant subjects consist of the mechanics of the allogenetic materials entering into the acceptor and they being to be recombined. The 3 effective changes include from studying morphology to genetic laws, from researching M1 generation to the next generations, from determining the single character to the synthetic traits. The mediated evidences of 4 aspects come from morphology, physiology and biochemistry, molecule biology. The 5 biological characteristics are mainly reproduction, development, photosynthesis, bad condition resistant and quality.     

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。