离子束辐照拟南芥生物学效应及其分子机理研究进展

综述了离子束辐照拟南芥种子生物学效应、 生长发育变化、 向重力性等研究的最新进展。 阐释了离子束辐照拟南芥染色体DNA碱基变化、 DNA断裂或损伤、 染色体重组、 突变遗传性等分子机理。 探讨了离子束介导外源基因转化拟南芥的有效性和机理。 同时展望了辐照拟南芥分子机理研究中的辐射原初效应传递、 信号转导等其他机理研究及重离子辐射生物学效应的应用前景。 Newly research progresses were summarized in effect of ion beams on seed surface, biological effect, growth, development, gravitropism and so on. Furthermore, mutation molecular mechanism of Arabidopsis thaliana was discussed, for example, alteration of DNA bases, DNA damage, chromosomal recombination, characteristics of mutant transmissibility, etc. Meanwhile, the achievements of transferring extraneous gene to Arabidopsis thaliana by ion beams were reviewed in the paper. At last, the future prospective are also discussed here in mutation molecular mechanism and the potential application of biological effect of heavy ion beams.     

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.     

MeV能量离子辐照拟南芥干种子和含水种子存活曲线对比

离子束辐照植物种子所产生的水自由基对植物种子存活率会有影响, 实验中为了观察这种辐射效应, 采用6.5 MeV能量的质子分别辐照拟南芥干种子和含水种子。 这个能量点的质子无论对干种子还是含水种子都可以完全穿透, 减少了因损伤部位不同带来的差异。 实验中采用的质子注量从4×109 ions/cm2到1×1014 ions/cm2。 实验结果显示干种子和含水种子存活曲线均成肩形下降趋势,而且含水种子下降注量点明显低于干种子。 这是自由基作用结果。 同时在辐射生物学靶学说的基础上, 建立了模型, 通过参数的合理设置,很好地拟合了实验数据。 The dry and water saturated seeds of Arabidopsis thaliana were irradiated by H+ions with 6.5 MeV in atmosphere. The ion fluence used in this experiment was in the range of 4×109 —1×1014 ions/cm2. According to the structure of the seed and TRIM simulation, the ions with the energy of 6.5 MeV can penetrate the whole seed. The experiment shows that the fluence response curves for the dry seeds and water saturated seeds had distinct shoulders and reduced rapidly. The experimental results show that the water imbibed seeds were more sensitive than the dry seeds and the reason is from free radicals reaction. A model has been constructed, and primely simulates the experiment data.     

和厚朴酚对非小细胞肺癌细胞的辐射增敏效应

研究了和厚朴酚（HNK）对非小细胞肺癌（NSCLC）细胞系A549和H1299对低线性能量转移（LET） X射线和高LET碳离子的辐射增敏效应。首先用CCK-8检测了HNK对A549和H1299细胞的生长抑制情况,发现20 μmol/L的HNK处理对细胞的生长抑制作用较弱。用该浓度HNK预处理细胞2 h后给予不同剂量X射线或碳离子的照射,克隆存活法检测细胞的辐射敏感性,Annexin-PI双染法检测细胞凋亡,γH2AX焦点法检测DNA的双链断裂（DSB）损伤。实验结果显示:与X射线相比,NSCLC细胞对碳离子更敏感,HNK预处理仅对碳离子照射有辐射增敏作用;与碳离子单独照射相比,HNK预处理联合碳离子照射诱导了更明显的细胞凋亡;在照射后24 h,HNK预处理联合碳离子照射引起的细胞γH2AX焦点阳性率维持在较高水平,而X射线照射没有这些效应。实验结果表明,HNK预处理抑制了NSCLC细胞DNA的DSB修复,诱导了细胞凋亡的发生,从而提高了细胞对碳离子的辐射敏感性。The radiosensitizing effect of Honokiol (HNK) on non-small cell lung carcinoma (NSCLC) cell lines A549 and H1299 to low-linear energy transfer (LET) X-rays and high-LET carbon ions was investigated in this study. First, the inhibitory effects of HNK on the growth of A549 and H1299 cells were detected by CCK-8 assay, and 20 μmol/L HNK treatment was found to induce a growth inhibitory effect slightly in these two cell lines. Cells were pre-treated with HNK and then irradiated with X-rays and carbon ions of different doses. Cellular radiosensitivity, apoptosis and DNA damage were analyzed by clonogenic survival, Annexin-PI staining and γH2AX foci, respectively. The results showed the cells were more sensitive to carbon ion irradiation compared to X-rays and the radiosensitization of HNK was only observed after carbon ion irradiation. Furthermore, the co-treatment led to higher apoptosis rate 48 h after irradiation and increased the positive rate of γH2AX foci 24 h after irradiation in A549 and H1299 cells compared with those in the groups treated with carbon ion irradiation alone. These phenomena were not observed after X-ray irradiation. Our data suggest that the pre-treatment with HNK inhibited DNA DSB repair, induced apoptosis and then enhanced the cellular radiosensitivity to carbon ions in NSCLC cells.     

12C6+离子束辐照菘蓝干种子当代效应

利用兰州重离子研究装置(HIRFL) 提供的12C6+ 离子束辐照菘蓝干种子(辐照剂量为10,35,60,90 和140 Gy,剂量率20 Gy/min),探讨了重离子束辐照对菘蓝M1代的生物学效应。研究发现,不同剂量的12C6+ 离子束辐照后,菘蓝种子的发芽率、成苗率、株高、根长和根冠比等生物学性状以及对菘蓝中靛玉红和4(3H) 喹唑酮含量均发生了变化,其中株高和根长随辐照剂量的增加而降低;菘蓝叶和根中的4(3H) 喹唑酮和靛玉红的含量随辐照剂量增加呈马鞍形增加关系。这表明:12C6+ 重离子束辐照菘蓝种子具有明显的当代损伤效应, 并可显著提高菘蓝中靛玉红和4(3H) 喹唑酮的含量,其辐照适宜诱变剂量为35 Gy。To investigate the M1 biological effects of heavy ions on Isatis indigotica Fort, its dry seeds were irradiated by 12C6+ beam with the dose of 0, 10, 35, 60, 90 and 140 Gy respectively,at the rate of 20 Gy/min delivered by the Heavy Ion Research Facility in Lanzhou (HIRFL). The results showed that biological characters such as germinating rate, germinating potential, survival rate, plant height, root height and root-shoot ratio were changed after irradiation. Moreover, the plant height and root height decreased in a dos dependent manner. The indirubin and 4(3H) quinazolinone content of Isatis indigotica Fort was improved and exhibited obviously “saddle” trends with irradiation dose increasing.Data suggest that exposure with low-dose 12C6+ to seeds of Isatis indigotica Fort has obvious injury effects at the first generation, and the active ingredient content of Isatis indigotica Fort may be improved by carbon ion beamirradiation. It is concluded that the suitable irradiation dose of mutation breeding is 35 Gy for the seeds of Isatis indigotica Fort.     

Heavy-Ion-Induced Sucrose and L-α-Alanine Radicals Investigated by EPR

We investigated the details of sucrose and L-α-alanine radicals produced by heavy-ion irradiation with various linear energy transfer (LET) values. The impact of the heavy ions on samples produced stable free radicals, which were analyzed by electron paramagnetic resonance (EPR). The spectral patterns obtained were similar among those for the irradiation of carbon (C), neon (Ne), silicon (Si), and argon (Ar) ions. The radical production of irradiated sucrose and alanine samples showed a linear relation with the absorbed dose. The total spin concentration obtained by heavy-ion irradiation decreases as logarithmic function of the LET. Quantitative EPR analysis showed that the production of sucrose and alanine radicals depended on both the ion species and the LET values under the same dose. Moreover, the radical-production cross sections for sucrose and alanine irradiated with C ions (290 MeV/u) were about 1.3 · 10⁻⁸ and about 3.0 · 10⁻⁹ μm², respectively. The results of the cross section suggest that each molecule becomes a stable radical with the interaction of several particles. Authors' address: Kouichi Nakagawa, Radioisotope Research Center, Fukushima Medical University, 1 Hikari-gaoka, Fukushima 960-1295, Japan     

重离子束对北里链霉菌的诱变效应

考察不同剂量重离子束对北里链霉菌的致死及诱变效应, 可确定最有利于筛选高产菌株的重离子束辐照剂量。 利用不同剂量的重离子束辐照北里链霉菌孢子, 统计了存活率、 致死率、 正负突变率。 结果发现, 在5 Gy重离子辐照时北里链霉菌出现较高致死率, 其后随剂量升高, 致死率变化较平缓。 各辐照剂量下正负突变率相比较, 40 Gy时正突变率最高, 负突变率相对较低, 存活率为0.92%。 因此确定40 Gy是对北里链霉菌高产菌株筛选最有利的辐照剂量。 To define the optimum dose of heavy ion beams for selecting high productive strains, we should study mortality and mutation effects of Streptomyces kitasatoensis irradiated by heavy ion beams in diffe rent doses. In this research, spores of Streptomyces kitasatoensis were irradiated by heavy ion beams with different doses. And survival rate, mortality rate, positive mutation and negative mutation were analyzed statistically. The results showed that high mortality rate appeared from 5 Gy and then the mortality rate curve became gently. Compared the positive and negative mutations in different doses, highest positive mutation was obtained in 40 Gy, while the negative mutation was lower in this dose, and the survival rate was 0.92%. So we defined that optimum dose of heavy ions radiation for Streptomyces kitasatoensis selection was 40 Gy in this experiment.     

重离子束用于肿瘤放射治疗的基础理论

通过与低传能线密度辐射治疗对比分析建立了重离子束肿瘤放射治疗的基础理论,提出了一些提高重离子束放射治疗疗效和减少对正常组织损伤的技术措施,旨在为已开展的重离子治癌临床研究提供理论依据． The basic theory of tumor radiotherapy with heavy ion beam was introduced in contrast to low LET irradiation therapy. Some useful methods are also suggested to improve the curative effect of heavy ion therapy and to spare the normal tissue around the tumor.     

碳离子束辐照处理对牛至精油微胶囊的缓释特性及抗鸡球虫病活性的影响

利用不同剂量的碳离子束辐照处理牛至精油微胶囊,研究对其缓释特性及抗鸡球虫病活性的影响。研究结果表明,通过碳离子束处理的牛至精油微胶囊,其主要活性成分百里香酚和香芹酚含量未发生变化,辐照处理后反应收率及产品的生物活性均提高,其中400 Gy剂量处理的牛至精油微囊的收率和生物活性均最高,收率达68.44%。通过对感染柔嫩艾美耳球虫的小鸡喂食后,其盲肠和十二指肠肿胀明显减轻,血液性内容物明显减少,血便指数在感染的第七天已降为0,抗球虫指数（ACI）达到179.39。The purpose of this study is using different doses of carbon ion beam irradiation on oregano essential oil microcapsules, to investigate function of control release and anticoccidial activity in chickens. The results showed that the content of main components in oregano essential oil microcapsules after carbon ion beam irradiation, thymol and carvacrol, have little change. Moreover, it can obtain more object products and better bioactivity after the irradiation treatment. Among these different doses of irradiation, our findings showed that Microcapsule groups with 400 Gy dose irradiation showed the highest yield (68.44%) and the best anticoccidial activity. After feeding chickens infected with Eimeria tenella, the swelling of caecum and duodenum were significantly mitigated and blood contents were also significantly reduced. The bloody stool index of chickens was as low as zero on the seventh day of infection, and the anticoccidial index (ACI) is 179.39.     

12C6+离子束辐照黄花蒿干种子当代生物学效应

利用12C6+重离子束对黄花蒿干种子辐照后,利用发芽率、根长、下胚轴长和株高4个指标研究不同剂量12C6+离子束辐照对黄花蒿的生物学效应。结果表明,随着辐照剂量的增加,黄花蒿的存活率和根长都呈现逐渐降低的趋势,而下胚轴长先上升后下降,株高总体呈现下降的趋势,但在60和90 Gy同时存在矮化与增高的植株。利用SRAP （sequence related amplified polymorphism）技术对辐照后M1代的黄花蒿植株进行研究,结果表明对照组和处理组之间的差异体现为特异性条带的变化。综合分析表明,12C6+的重离子辐照黄花蒿干种子可以产生显著的当代损伤效应,同时引起植物DNA的多态性变化。     

~(12)C~(6+)离子束与电子束辐照对薰衣草当代诱变效应的比较

采用100 MeV的~(12)C~(6+)离子束和1.2 MeV的电子束辐照薰衣草干种子,研究了2个品种的薰衣草干种子对不同辐射源的辐照生物学效应,以期找到薰衣草干种子的最佳诱变参数。结果表明:不论是薰衣草701还是702,经~(12)C~(6+)离子辐照后,其发芽率表现为先增大后减小的趋势;经电子束辐照后,发芽率随着剂量的升高而降低,电子束辐照后的发芽率要低于~(12)C~(6+)离子束。另外,2个品种的薰衣草经~(12)C~+离子辐照后,其胚轴胚根长度以及幼苗鲜重也较电子束辐照后的大。由此可知,重离子的辐照效果优于电子束的,有利于后期筛选出新的薰衣草突变株。     

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。     

~(12)C~(6+)离子束辐照紫苏干种子当代效应

利用兰州重离子研究装置(HIRFL)提供的12C6+离子束辐照紫苏干种子(辐照剂量为40,80和120Gy,剂量率4Gy/min),探讨了重离子束辐照对紫苏M1代的生物学效应。结果发现,经不同剂量的12C6+离子束辐照后,紫苏种子的发芽率、发芽势、存活率、株高、分枝数、单株产量和千粒重等生物学性状均发生了变化,其中发芽势、单株产量和千粒重随辐照剂量的提高而降低,且有明显的剂量效应关系,但发芽率、大田成活率、株高和分枝数却随辐照剂量的增大,呈现出明显的"抛物线"趋势;紫苏幼苗根尖细胞的微核率和染色体畸变率随辐照剂量增加呈线性增加关系。这表明:12C6+重离子束辐照紫苏种子,具有明显的当代损伤效应,在本试验剂量范围内,低剂量辐照对发芽率和成活率有促进作用。     

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。     

重离子加速器示范装置被动式束流配送系统次级中子特征的蒙特卡罗研究

在碳离子放射治疗中,碳离子束在剂量配送过程中会与束流输运线相互作用,形成以中子辐射为主的外辐射场.由于中子是高LET射线,具有较高的相对生物学效应,减少碳离子放疗中产生的次级中子有助于降低放疗后正常组织并发症几率及二次肿瘤风险.利用蒙特卡罗方法对保守情况(能量为400 MeV/u,多叶光栅完全闭合)下碳离子治疗被动式束...     

Time-course of aberrations and their distribution: impact of LET and track structure

The biological response to high linear energy transfer (LET) radiation differs considerably from that to low LET radiation and this has been attributed to differences in the spatial energy deposition of both radiation qualities. In the case of X-rays the energy is deposited uniformly within the cell nucleus and produces damages in a purely stochastic manner. In contrast, for particles the energy is deposited inhomogeneously along the ion trajectory and the local dose decays with the square radial distance from the center of the track. This nonuniformity affects the yield and the distribution of aberrations among cells. Moreover, after high LET exposure a relationship between the aberration yield and cell cycle delay was observed. In this study, we present a detailed analysis of the distribution of aberrations in human lymphocytes reaching mitosis at early and later times after low and high LET exposure. Aberration data were fit to stochastic distributions demonstrating that the delay is related to the number of particle traversals per cell nucleus. To further elucidate this relationship, we introduce a Monte Carlo phenomenological model which incorporates the number of particle hits per nucleus. This value was derived by fitting theoretical distributions to the experimental data. Additionally, the probability that a cell traversed by a particle reaches mitosis at a given time was calculated. The analysis of biological data and numerical simulations clearly show the impact of the track structure on the formation of chromosome aberrations and their distribution among cells.     

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.