DNA双链断裂产额的新算法

DNA双链断裂(DSB)的定量分析是高传能线密度辐射生物学效应机理研究的重要手段.从实际应用出发,推导了一个新的计算公式———平均分子量法.该法不探究DSB片段的具体分布模式,实际操作中却又包含了片段的含量和分布;而且形式简单、操作简便,尤其适合于荧光扫描数据. Quantitive analysis of DNA double strand break (DSB) is of importance for studying on the mechanisms of radiobiological effects induced by high linear energy transfer(LET) irradiation. For the practical use, one new calculating formula named the method of average molecular weight is deducted. In this method, the factual distribution of DSB fragments is not considered, but actually, the content and distribution of DSB fragments are included. Totally, this method is simple in shape and easy to use, esp...     

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.     

重离子诱导的质粒DNA双链断裂分布研究

利用能量为7．2MeV／u氖离子束辐照体外质粒DNA:pUC18，采用恒场凝胶电泳结合多功能荧光成像系统研究了pUC18双链断裂片段的分布。证实了双链断裂片段分布的非随机性，结果还发现DNA断裂后片段的交联现象，而且交联片段的分布也是非随机的。DNA is considered to be the most important and sensitive target in biological systems. In addition to the base damage, DNA strand breaks are the major lesion in the genome due to exposure to ionizing radiation. Mutation can be introduced to DNA as a result of enzymatic processing of DNA lesions or post irradiation replication. However, the mechanisms of radiation induced mutations are not well clarified at the molecular level. To study the effect on the simple plasmid DNA of heavy ion is even predominant or more feasible. Plasmid pUC18 DNA was prepared and irradiated by neon beam (7.199 MeV/u). The fragment distributions were determined by quantifying the ethidium bromide fluorescence. It can be seen that the shape of the intensity distributions is vastly different for the used radiation Dose. The distribution produced shows an excess of fragments particularly in 3 000 and 10 000 Gy the size range between 20—40 kbp and 20—50 bp. This clustering of double stranded fragments might be influenced by the higher order chromatin structure of genomic DNA. If so, DNA loop structures could correspond to the size range for which we observed DSB clustering. Further studies aim at elucidating the heterogeneity of DSB induction within the genome and investigate the influence of chromatin structure on the non random fragment distribution.     

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.     

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.     

Molecular dynamics simulation of latent track formation in α-quartz

The latent ion track in α-quartz is studied by molecular dynamics simulations. The latent track is created by depositing electron energies into a cylindrical region with a radius of 3nm. In this study, the electron stopping power varies from 3.0keV/nm to 12.0keV/nm, and a continuous latent track is observed for all the simulated values of electron stopping power except 3.0keV/nm. The simulation results indicate that the threshold electron stopping power for a continous latent track lies between 3.0keV/nm and 3.7 keV/nm. In addition, the coordination defects produced in the latent track are analyzed for all the simulation conditions, and the results show that the latent track in α-quartz consists of an O-rich amorphous phase and Si-rich point defects. At the end of this paper, the influence of the energy deposition model on the latent track in α-quartz is investigated. The results indicate that different energy deposition models reveal similar latent track properties. However, the values of the threshold electron stopping power and the ion track radius are dependent on the choice of energy deposition model.     

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.     

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.     

Interaction of O3+ Ions with Al2O3 Nanocapillaries

We investigate angular distributions of the transmitted 60 keV and 120 keV O^3＋ ions through nanocapillaries with 50hm diameter and 10μm length. The experimental results are in agreement with the ＇guiding effect＇ found by Stolterfoht et al. For different tilted angles of the nanocapillary membrane with respect to the beam, the angular distributions of the transmitted ions are measured by a one-dimensional micro-channel plate （MCP） detector. Moreover, the dependence of this guiding effect on the projectile energy is studied.     

Stark Deceleration of an Effusive Molecular Beam by a Single Semi-Gaussian Beam

We propose a novel scheme to decelerate a cw precooled molecular beam by using an optical Stark decelerator, which is composed of a red-detuned cw semi-Gaussian beam. The dynamical process of the optical Stark deceleration for a cw methane molecular beam is studied by Monte Carlo simulations. It is realized that the proposed optical Stark decelerator can be used to continuously slow a cw methane molecular beam with a longitudinal temperature of 30mK, and a maximum reduction in the most probable speed of 0.388m/s （corresponding to a relative change of 9.93%） can be obtained by using a single semi-Gaussian beam with a power of 500W and a maximum central intensity of about 7.96×10^7 W/cm^2.     

不同LET辐射致DNA集簇性损伤的研究

利用辐照质粒DNA构象变化的分子模型,以DNA糖苷酶Fpg和AP核酸内切酶EndoIII识别并切割辐射所致DNA碱基损伤,将其转换为DNA断裂损伤,通过电泳分析DNA分子构象变化,研究比较γ射线、质子和7Li离子诱发DNA集簇性损伤。50Gy以上高剂量γ辐射对质粒DNA的损伤主要表现为单链断裂(SSB)和很少比例的双链断裂(DSB),并能产生一定水平的集簇性损伤。相比之下,高能质子束和高LET的7Li离子直接所致DNA的断裂损伤以及所产生的集簇性碱基损伤比γ射线的要严重,质子10Gy照射就可诱发明显的集簇损伤。     

Novel image processing interface to relate DSB spatial distribution from experiments with phosphorylation foci to the state-of-the-art models of DNA breakage

We apply new image processing tools and computer modeling of DSB formation to analyze new experimental data on DSB (DNA double-strand break) yield (#DSB per base pair per Gray). There is LET-dependent DSB clustering within the nucleus volume, and clustering of DSBs along the DNA length, which we show through modeling the whole set of human chromosomes. In recent experiments, DSBs are imaged as phosphorylation sites of the histone protein H2AX, denoted as γH2AX foci, and it is suggested that foci images indicate the spatial distribution of DSBs. For high-LET radiation, DSBs should be located closer to the track center or grouped around ionization sites leading to clustered DSBs as described theoretically, and as can be seen in images. We describe the successful segmentation of foci images and determine foci statistics that will relate models of DSB spatial correlation and clustering along DNA length to the experimental data. The foci data can be used to analyze high-LET effects on DNA fragment sizes and DSB distributions.     

The influence of DNA inhibitor synthesis on the induction and repair of double-strand DNA breaks in human lymphocytes under action of radiation with a different linear energy transfer

The influence that inhibitors of repair and replicative DNA synthesis, 1-β-D-arabinofuranosyl-cytosine and hydroxyurea, have on the formation and repair kinetics of double-strand breaks (DSBs) in peripheral human blood lymphocytes under the influence of radiation with a different linear energy transfer (LET) (gamma quanta and accelerated heavy ions) is studied. It is demonstrated that lithium and boron ions with LETs of 20 and 40 keV/μm, respectively, possess higher biological effectiveness with respect to the DNA DSB induction criterion. The value of the relative biological effectiveness of accelerated lithium and boron ions is 1.5 ± 0.1 and 1.6 ± 0.1, respectively. It is found that, upon cell irradiation by gamma quanta in the absence of inhibitors, efficient DNA DSB repair is observed during incubation. Under the conditions of cell incubation and in the presence of inhibitors, some growth in the number of DNA DSBs, rather than a reduction, is observed after 5-h incubation. In the case of the action of accelerated boron ions (as well as gamma quanta), under normal conditions, the efficient repair of induced DNA lesions takes place. Unlike the action of gamma quanta, in the case of cell incubation in the presence of radiomodifiers, the number of induced DNA DSBs falls. These results may testify to the fact that the repair of double-strand DNS breaks takes place under the action of ionizing radiation with a different LET on mammalian cells in the presence of DNA synthesis inhibitors Ara-C and HU. It is concluded that, for cells subject to gamma irradiation, no DNA DSB repair is observed due to the large contribution of single-strand incision DNA breaks formed in the postradiation period in the course of excision nucleotide repair.     

In-situ study on thermal decomposition of 1,3-disilabutane to silicon carbide on Si(1 0 0) surface

The intermediates of thermal decomposition of 1,3-disilabutane (SiH₃CH₂SiH₂CH₃, DSB) to form SiC on Si(1 0 0) surface were in situ investigated by reactive ion scattering (RIS), temperature programmed reactive ion scattering (TPRIS), temperature programmed desorption (TPD), and auger electron spectroscopy (AES). DSB as a single molecular precursor was exposed on Si(1 0 0) surface at a low temperature less than 100 K, and then the substrate was heated up to 1000 K. RIS, TPD, and AES investigations showed that DSB adsorbed molecularly and decomposed to SiC via some intermediates on Si(1 0 0) surface as substrate temperature increasing. Between 117 and 150 K molecularly adsorbed DSB desorbed partially and decomposed to CH₄Si₂, which is the first observation on Si(1 0 0) surface, and further decomposed to CH₄Si between 150 and 900 K. CH₄Si lost hydrogen and formed SiC over 900 K.     

放疗污染电子对皮肤细胞损伤的蒙特卡罗模拟研究

采用Geant4-DNA低能物理模型研究了污染电子在细胞DNA水平上的物理作用和能量沉积。利用密度聚类算法分析了损伤产额分布;并结合皮肤细胞辐射敏感性参数和临床表征剂量,探讨了低能电子对皮肤细胞的损伤特征。模拟发现,产生的DSB中约20%是cDSB;DSB产额是SSB的约4%;损伤聚类包含的SSB一般3个,20 keV和100 keV电子也可造成5个SSB的聚类损伤;晚反应、高α值的组织更应注意防护低能电子辐射;由于LQ模型中剂量平方项的影响,cDSB损伤致死系数ε随入射电子数目的增加而增大。当105个电子入射时,ε的数值可较单个电子增大3%～15%。可通过调节直接电离损伤概率弥补间接损伤产额,研究细胞辐照损伤的内在机理。本工作建立的低能电子对细胞DNA的损伤模型及结果,可用于评价放疗中低能污染电子对皮肤细胞的损伤效应。     

Formation of Direct and Enzymatic DNA Double-Strand Breaks in the Presence of Repair Inhibitors after Exposure to Radiations of Different Quality

With the use of the DNA comet assay and immunocytochemical staining, regularities have been studied in the induction and repair of DNA double-strand breaks(DSBs) in human cells after exposure to ⁶⁰Co γ-rays and accelerated heavy ions with different linear energy transfer (LET) in the presence of the DNA repair inhibitors cytosine arabinoside and hydroxyurea. It is shown that for heavy ions the agents’ modifying effect decreases with increasing particles’ LET. The approach involving DNA synthesis inhibitors used in this study allows an estimation of the proportion of enzymatic DNA DSBs in total DSB yield after exposure to ionizing radiations of different quality.     

Fluorescence spectroscopy of potential electroluminescent materials: Substituent effects on DSB and segmented PPV derivatives

The absorption and fluorescence of substituted distyrylbenzene (DSB) derivatives and segmented poly(phenylene vinylene) (PPV) derivatives are characterized by long-wavelength absorption maxima and absorption coefficients of λ_a = 380–450 nm, ε = 20,000–60,000 M⁻¹ cm¹ and fluorescence maxima, quantum yields, and decay times of λ_r = 440–530 nm, Φ_f = 0.2–0.9, and Τ = 0.8–2.5 ns, respectively. Alkoxy substituents at the central phenylene ring of DSB groups increase the bathochromic shift in the spectra in comparison to DSB, without a significant decrease in the high DSB fluorescence quantum yield. Both phenyl and cyano substitutions at the vinylene bridge lead to a further bathochromic shift of the fluorescence and a decrease in the quantum yield to ca. 0.4. The DSB derivatives and the related segmented PPV derivatives show nearly the same absorption spectra, fluorescence spectra, and radiative rate constantsk _f= Φ_f/Τ, indicating the efficacy of the segmentation of the polymer chain. The radiative rate constants determined by the Φ_f and Τ values and by the Strickler/Berg formula are in reasonable agreement. This supports the possibility of interpreting the properties of the polymers in terms of their DSB units. The decrease in the emission anisotropy can be ascribed to multistep energy transfer processes between different oriented segments.     

双面阶梯埋氧层部分SOI高压器件新结构

提出了双面阶梯埋氧层部分绝缘硅(silicon on insulator,SIO)高压器件新结构. 双面阶梯埋氧层的附加电场对表面电场的调制作用使表面电场达到近似理想的均匀分布, 耗尽层通过源极下硅窗口进一步向硅衬底扩展, 使埋氧层中纵向电场高达常规SOI结构的两倍, 且缓解了常规SOI结构的自热效应. 建立了漂移区电场的二维解析模型, 获得了器件结构参数间的优化关系. 结果表明, 在导通电阻相近的情况下, 双面阶梯埋氧层部分SOI结构击穿电压较常规SOI器件提高58％, 温度降低10—30K. 关键词： 双面阶梯 埋氧层 调制 自热效应