12C6＋离子束辐照对大青叶生理生化特性的影响

利用辐射能量为100 MeV/u的12C6＋ 重离子束辐照大青叶种子,辐照剂量分别为20、40、50和80 Gy研究其对大青叶M1代的生物学效应.研究结果表明,随着辐照剂量的增大,大青叶的根系活力和叶绿素含量显著降低,发现辐照损伤主要抑制了根的生长及营养吸收;酚酸类化合物含量随着辐照剂量的增大而升高的趋势,表明辐照可提高大青叶的抗氧化和抗病虫害能力;可溶性糖、丙二醛、超氧化物歧化酶（SOD）和过氧化物酶（POD）含量变化的总体趋势为随着辐照剂量的增大先升后降.表明重离子辐照能改变大青叶的一些生理生化特征,其中40 Gy的12C6＋离子束辐照有利改善大青叶抗逆性和一些有效化学成分的积累.     

低剂量率下高线性能量转移碳离子束辐照人类唾液腺细胞的存活效应

利用日本千叶重离子医用加速器 HIMAC 提供的碳离子束,对人类唾液腺细胞 (HSG) 在剂量率为 0.5 Gy/h 的低剂量率条件下进行了辐照,运用标准的克隆形成法得到了 3 种不同剂量平均线性能量转移 (LET) 碳离子束辐照 HSG 细胞的剂量存活效应 . 与先前 HSG 细胞在治癌剂量率 (1~5 Gy/min) 下对相近剂量平均 LET 碳离子束辐照的剂量存活效应数据相比, HSG 细胞对高 LET 碳离子束辐射表现出明显的剂量率效应 . 为在相同条件下得到碳离子束对 HSG 细胞的相对生物学效应 (RBE) ,利用 60Co-γ射线在剂量率为 0.5 Gy/h 的条件下辐照了 HSG 细胞,得到该细胞系对低 LET 射线响应的剂量存活效应 . 与先前在治癌剂量率下得到的 RBE 值相比,低剂量率条件下得到的 RBE 值总体减小 . 由实验发现的剂量率效应及低剂量率条件下 RBE 值的减小,表明由高 LET 碳离子束造成的辐射损伤在低剂量率条件下也存在着显著的修复效应 . 据此,对辐射造成细胞致死的原因进行了探讨 .     

重离子束辐照牧草的细胞学研究

采用80MeV／u^20Ne^10 叶离子束贯穿处理豆科与禾本科牧草种子,从实验室种子萌发和根尖细胞的观测分析,随着贯穿剂量的增加,幼苗生长明显减弱,呈负相关性;而染色体总畸变率和徽核率随剂量的增加而显著增加,呈正相关性。结果表明：禾本科牧草比豆科牧草对重离子辐射敏感性强,禾本科牧草适宜剂量为20Gy～30Gy,豆科牧草辐照剂量应高于150Gy。     

UV-B辐射对拟南芥种子萌发和幼苗生长的影响

以哥伦比亚生态型(Columbia-0)拟南芥(Arabidopsis thaliana)为实验材料, 人工模拟UV-B辐照处理拟南芥种子, 统计其发芽势和发芽率, 测定根长、株高、叶绿素、可溶性糖、可溶性蛋白以及丙二醛(MDA)含量, 研究UV-B辐照处理对拟南芥种子萌发和幼苗生长的影响。研究结果表明, 低剂量的UV-B辐照可以促进拟南芥种子萌发和幼苗生长, 并且最佳辐照剂量为1.0 kJ·m^–2(P<0.05), 此时的发芽势、发芽率、根长、株高、叶绿素、可溶性糖及可溶性蛋白的含量均达到最大, 而丙二醛的含量变化则不明显(P>0.05)。当辐照剂量大于1.0 kJ·m^–2时, 促进作用逐渐变小, 并且随着辐照剂量的增加, 表现出了抑制作用。实验结果表明, 适当剂量的UV-B辐照在一定程度上可以促进拟南芥种子萌发和幼苗生长, 而过高剂量的辐照则对其产生抑制或损伤作用。     

离子束介导GUS基因转化拟南芥种子的研究

以拟南芥种子为研究对象,研究离子束介导GUS基因导入拟南芥种子的转化效果。结果表明:直接注入干种子后进行转化,未得到瞬间表达的结果。而注入玻璃化保护后的2天龄幼苗,再进行转化,转化效果明显提高,得到了大约10%的瞬间表达。揭示对于像拟南芥这样的小粒种子,采用幼苗注入后转化,效果可能会更好一些。     

氮离子束对小麦种子不同部位作用的突变效应

本文描述了荷能重离子束（^14氮^1 和^14氮^7 ）在作物改良上的应用。为了探讨离子束轰击小麦种子不同部位（例如：种皮,种胚和整粒种籽,包括胚乳、胚和种皮）后的不同反应,采用了不同能量的氮离子。轰击不同部位是通过改变离子能量来实现的。在这个研究中,我们选择了三种能量以达到轰击不同部位的目的,它们是超低能区的110keV,低能区的15．7MeV/u和中能区的72MeV/u.根据TRIM 91程序计算,它们在种子内的射程依次为0．44μm,0.61mm和9.6mm。所以,110keV的离子不能贯穿种皮,因为它的厚度72μm,只能极浅层注入种皮而不能触及胚细胞（称这种情况为轰击部位1）,15．7MeV/u的离子能够贯穿种皮并注入胚内（厚度约1mm),但不能进入胚乳（称这为轰击部位2）,72MeV/u的离子能从种子的胚部到顶部贯穿整个麦粒（麦粒长约7mm)（称这为轰击部位3）。上述三种能量的氮离子辐照了三个品种（定西24、88－12、82－579）的春小麦种子。而后进行了室内实验和大田培育,得到了50％出苗率时的剂量D50,统计了上述三个轰击部位下根尖细胞中的微核率及染色体畸变率,大田中产生了一些新的变异,例如增产（达百分之几十）,早熟（五天左右）,矮杆（低约20cm),抗（条锈）病,并且显示了轰击不同部位的突变频率与突变谱,还简略地讨论了三种情况的突变机理。     

离子注入法将外源DNA直接导入小麦的研究

通过离子束介导法将外源GUS基因直接导入小麦成熟种子.组织化学染色结果表明GUS基因的瞬时表达率可以达到70%以上.当代(Ru)PCR分析结果表明,阳性植株的频率与离子注入剂量有关,适宜的注入剂量为7×106离子/cm2.PCR-Southern和Southern B1ot分析结果表明外源基因已整合到小麦基因组中,说明离子束介导外源DNA直接导入小麦是可行的.另外还探讨了用离子束介导创造小麦远缘分子杂种的可能性.     

重离子及电子辐照沙打旺(Astragalus Adsurgens Pall)干种子的辐射生物学效应

本文采用80MeV/u12C6+和50keV的电子辐照沙打旺干种子,研究沙打旺干种子不同注量的辐照生物学效应,检测其生理生化指标。结果表明:无论是碳离子辐照还是电子辐照,发芽势都随注量的增加先升高后降低,发芽率都随注量增加逐渐降低。其次,碳离子辐照样品,其超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性变化随注量增加先升高后降低,即存在一个峰值,过氧化氢酶(CAT)在高于5×106ions/cm2注量的离子辐照中其活性基本不发生变化;不同注量的电子辐照,样品的CAT、SOD、GSH-Px活性都是先升高后降低。第三,无论是碳离子辐照还是电子辐照样品的谷胱甘肽(GSH)含量变化不显著,与注量的变化没有明显的相关性。总而言之,本工作发现碳离子及电子辐照样品,其多个生理生化指标均随注量的增加呈先升高后降低的趋势,但GSH变化对碳离子或电子辐照都不敏感。     

沿重离子贯穿深度的番茄干种子辐射生物学效应研究

本文采用80MeV/u的^12C^6 辐照番茄干种子,研究处于不同离子贯穿深度番茄干种子的辐照生物学效应,检测其生理生化指标。结果表明：辐照后不同贯穿深度上番茄种子的发芽势与物理剂量对应,第8层样品（对应于碳离子在水中的贯穿深度为15mm）出现峰值76.7％,发芽率随离子入射深度的变化不明显;不同贯穿深度辐照样品的根尖细胞微核率也与物理剂量相对应,在第6、7层样品中出现峰值0.257％;另外,辐照样品幼苗茎叶的一氧化氮合酶（NOS）活性、谷胱甘肽（GSH）含量及总抗氧化力在第9层出现峰值,谷胱甘肽过氧化物酶（GSH-PX）活性在第7-8层出现峰值,超氧化物歧化酶（SOD）、过氧化氢酶（CAT）活性的变化与重离子贯穿深度没有明显的相关性。总而言之,本工作发现辐照品的多个生理生化指标均与入射离子的微剂量分布相对应,即存在生物效应峰。     

80MeV/u20Ne10+离子对SMMC-7721细胞DNA损伤及细胞周期的影响

从辐照剂量和修复时间两个角度研究了重离子辐照对肿瘤细胞DNA损伤及细胞周期的影响,为重离子治癌的临床应用积累基础数据。不同剂量的80MeV／u^20Ne^10 辐照SMMC—7721细胞样品,利用单细胞凝胶电泳技术(Single Cell Gel Electrophoresis,SCGE)对细胞DNA损伤进行了检测,利用流式细胞技术(Flow Cytometry Methods,FCM)对细胞周期变化进行了分析。80MeV／u^20Ne^10 辐照后4小时内,SMMC—7721细胞的DNA损伤与辐照剂量呈线性关系,在0小时组其线性相关因子r为0．9621,4小时组为0．914;随着修复时间的增加,DNA损伤与辐照剂量不再线性相关,但0．5Gy,1Gy和2Gy三个剂量点的DNA损伤程度极为相近。另外,重离子辐照后SMMC—7721细胞发生S期和G2／M期阻滞现象,其随剂量变化及时间变化的规律不同于X、γ等低LET(Linear Energy Transfer)射线辐照。     

Biological effects of ion beams in Nicotiana tabacum L.

The biological effects of ion beams on Nicotiana tabacum L., particularly the induction of chromosome aberrations, were investigated. Dry seeds were exposed to ¹²C⁵⁺, ⁴He²⁺ and ¹H⁺ beams with linear energy transfer (LET) ranging from 1 to 111 keV/μm and irradiated with gamma-rays. Ion beams were more effective in reducing germination and survival of the seeds than gamma-rays. The LD₅₀ for ¹²C⁵⁺ beams, ⁴He²⁺ beams and gamma-rays were 35, 60 and 500 Gy, respectively. The frequencies of mitotic cells with chromosome aberrations, such as chromosome bridges, acentric fragments and lagging chromosomes in the root tip cells of the exposed seeds, increased linearly with increasing doses. Relative biological effectiveness (RBE) values, based on the doses that induced a survival inhibition of 50% and a 10% frequency of aberrant cells, were 14.3–17.5 for the ¹²C⁵⁺ beams, 7.0–8.3 for the ⁴He²⁺ beams and 7.8 for the ¹H⁺ beams. Furthermore, the relative ratios of the chromosome aberration types were significantly different between the ion beam and the gamma-ray regimes: chromosome fragments were more frequent in the former, and chromosome bridges in the latter. Based on these results, we concluded that the repair process of initial lesions induced by ion beams may be different from that induced by low- LET radiation. Received: 29 October 1998 / Accepted in revised form: 25 March 1999     

LET and ion-species dependence for mutation induction and mutation spectrum on hprt locus in normal human fibroblasts.

We have been studying LET and ion species dependence of RBE in mutation frequency and mutation spectrum of deletion pattern of exons in hprt locus. Normal human skin fibroblasts were irradiated with heavy-ion beams, such as carbon- (290 MeV/u and 135 MeV/u), neon- (230 MeV/u and 400 MeV/u), silicon- (490 MeV/u) and iron- (500 MeV/u) ion beams, generated by Heavy Ion Medical Accelerator in Chiba (HIMAC) at national Institute of Radiological Sciences (NIRS). Mutation induction in hprt locus was detected to measure 6-thioguanine resistant colonies and deletion spectrum of exons was analyzed by multiplex PCR. The LET-RBE curves of mutation induction for carbon- and neon-ion beams showed a peak around 75 keV/micrometers and 155 keV/micrometers, respectively. On the other hand, there observed no clear peak for silicon-ion beams. The deletion spectrum of exons was different in induced mutants among different ion species. These results suggested that quantitative and qualitative difference in mutation occurred when using different ion species even if similar LET values.     

N+离子注入对大豆种子活力及其幼苗的抗氧化酶活性影响

本文研究了用25keV N+注入丰豆103的种子后,N+离子对其种子的活力及子叶伸展后48小时和96小时的幼苗内蛋白质含量、谷胱甘肽巯基转移酶(GSH-Ts)、谷胱甘肽过氧化酶(GSH-Px)、抗坏血酸过氧化物酶(ASA-POD)活性的影响.结果表明当N+注量在2.6×1016N+/cm2～5.2×1016N+/cm2时,种子的发芽率、发芽指数、活力指数都明显提高;幼苗的可溶性蛋白含量高于对照.在6.5×1016N+/cm2～10.4×1016N+/cm2注量时,幼苗的可溶性蛋白含量低于对照,96小时幼苗可溶性蛋白的含量高于48小时,说明辐射引起的损伤可随生长时间的增加而有所恢复.高注量可引起幼苗内一些抗氧化酶活性的升高,且随注量的增加酶的活性升高也越明显,96小时幼苗的GSH-Px和ASA-POD活性高于48小时幼苗,GSH-Ts活性略有下降.而低注量(1.3×1016N+/cm2～5.2×1016N+/cm2)的上述酶指标升幅不大.说明经N+离子处理后可通过诱导这些抗氧化酶活性的升高起到减轻伤害的作用.     

Chromatid-type aberrations following irradiation in G0 lymphocytes with heavy ions

We describe a low level of chromatid-type aberrations which included the relatively rare isochromatid/chromatid triradial in peripheral blood lymphocytes that were irradiated, ostensibly in G0, with accelerated heavy (12)C ions. These were produced only at the energies of 69 MeV/n (34.6 keV/microm), almost absent at the energy of either 58.6 MeV/n (46.07 keV/ microm) or 19.3 MeV/n (97 keV/microm), nor were they found after low-LET X-rays. Mechanisms potentially responsible for their formation are discussed.     

Irradiation of mammalian cultured cells with a collimated heavy-ion microbeam

As the first step for the analysis of the biological effect of heavy charged-particle radiation, we established a method for the irradiation of individual cells with a heavy-ion microbeam apparatus at JAERI-Takasaki. CHO-K1 cells attached on a thin film of an ion track detector, CR-39, were automatically detected under a fluorescence microscope and irradiated individually with an 40Ar13+ ion (11.5 MeV/nucleon, LET 1260 keV/microm) microbeam. Without killing the irradiated cells, trajectories of irradiated ions were visualized as etch pits by treatment of the CR-39 with an alkaline-ethanol solution at 37 degrees C. The exact positions of ion hits were determined by overlaying images of both cells and etch pits. The cells that were irradiated with argon ions showed a reduced growth in postirradiation observations. Moreover, a single hit of an argon ion to the cell nucleus resulted in strong growth inhibition. These results tell us that our verified irradiation method enables us to start a precise study of the effects of high-LET radiation on cells.