X射线辐照引起的线粒体D310片段突变的快速检测

采用4 Gy X射线辐照人乳腺癌细胞MCF 7, 分别在照射后0, 2, 4, 8, 16, 24, 48, 72和144 h时间点收集细胞, 提取DNA并以聚合酶链式反应进行扩增, 再以BsaXI限制性内切酶对扩增产物进行消化, 消化产物以琼脂糖凝胶电泳进行分离。 结果表明, 4 Gy X射线辐照可引起D310片段突变, 且该突变在经辐照之后144 h检测水平明显超过野生型。 Human breast cancer cell line MCF 7 was irradiated with 4 Gy X ray, collected at 0, 2, 4, 8, 16, 24, 48, 72, 144 h after irradiation, respectively. Whole genome DNA including mtDNA were extracted at each time point, and amplified by polymerase chain reaction(PCR). Then the PCR product was subjected to BSAXI digestion, all of digestion product then underwent a brief electrophoresis. Results showed D310 mutation can be induced by 4 Gy X ray irradiation and D310 mutation can overwhelm the normal phenotype 144 h after irradiation.     

X射线致HeLa细胞损伤过程中NADPH氧化酶的作斥

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,12Cry 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p47^phox在细胞质积聚并和细胞膜亚基gp91^phox结合;Western blotting检测结果显示,NADPH氧化酶的关键亚基gp91^phox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。     

X射线辐射与模拟微重力对K562细胞红系分化的联合效应及机制研究

使用氯化高铁血红素(hemin)诱导K562细胞分化作为红系分化模型,并对其进行X射线辐照及地面模拟微重力效应处理,研究辐照、微重力及二者的复合效应对红系分化的影响及机制。X射线辐照及模拟微重力效应处理后的联苯胺染色阳性率及CD235a蛋白表达均下调,细胞增殖抑制、细胞凋亡率及坏死率均增高,红系相关转录因子EPOR及GATA-1基因表达下调,且X射线辐照及模拟微重力效应的联合作用效应强于两者单独作用。辐照及微重力联合处理影响细胞中PI3K复合物调控亚基PIK3R2基因表达,加入PI3K抑制剂3-MA后细胞凋亡率及坏死率增高、红系分化率降低。以上结果表明,X射线辐照及模拟微重力效应对红系分化具有协同抑制作用,其机制与红系相关转录因子EPOR、GATA-1及生长信号通路因子PI3K相关。     

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。     

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.     

~(60)Co-γ射线茶穗辐照及其抗辐照性研究

为了探究~(60)Co-γ射线辐照对茶树插穗的诱变效应,选取广西区内特有品种尧山秀绿、桂香18号、桂绿1号的插穗,分别用2.5Gy、9.0Gy、15.0Gy的~(60)Co-γ射线进行辐照处理。探索辐照对茶树发芽率、存活率等的影响,为广西区内茶树辐照育种提供有效的途径和方法。实验表明:自养期内,三个品种中桂绿1号的抗辐照性能最佳,一定剂量(9.0Gy)的辐照还能在一定程度上减小桂绿1号的落叶率;茶树的插穗对辐照很敏感,2.0Gy的低吸收剂量的辐照已经明显阻碍插穗的生根和发芽;茶树的抗辐照性能因品种不同而有明显的强弱差异,每个品种的适合辐照剂量不相同;依据后期存活率,桂香18号的抗辐照性能比尧山秀绿和桂绿1号更强,同时,桂香18号在9.0Gy条件下的抗辐照能力尤为突出。     

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.     

锆英石的抗γ射线辐照能力和Rietveld结构精修

为研究放射性核素固化介质备选矿物锆英石的抗γ射线辐照结构稳定性,以澳大利亚锆英石为研究对象,通过⁶⁰Co源γ射线辐照装置对样品施以1728 kGy的γ射线辐照.利用X射线荧光光谱仪、扫描电子显微镜和X射线衍射仪对样品的元素含量、γ射线辐照前后的微观形貌及物相变化进行表征,同时利用Rietveld方法对γ射线辐照前后的样品进行了结构精修.结果表明:澳大利亚锆英石经1728 kGy剂量的γ射线辐照后未发生物相变化,射线辐照前后样品的晶胞参数仅发生了10^-4 量级的变 关键词： 锆英石 γ射线 辐照 Rietveld结构精修     

X射线和~(60)Coγ射线辐照食管癌细胞的拉曼光谱研究

食管癌细胞(EC9706)经不同剂量X射线和60Coγ射线辐照后继续培养24h,利用激光拉曼光谱分析EC9706细胞内部蛋白质、核酸、脂类等生物大分子的构象和含量变化。分析发现,两种放射源各剂量组拉曼光谱的峰强和频移与空白对照组之间都存在较大的差异。主要表现为(1)X射线辐照后,对照组光谱中存在的某些谱带,个别剂量组中该谱带却消失。蛋白质主链中骨架C-N振动引起的1114 cm-1谱带在各剂量组中普遍消失,膜脂中膜结合β-胡萝卜素的C=C振动谱带1523 cm-1仅存在于6Gy组。(2)60Coγ射线辐照后,蛋白质酰胺Ⅲ带在中等剂量(4、5Gy)组中β折叠结构向无规卷曲转化,大剂量组中DNA中的磷酸二酯(O-P-O)基团的非氢键化程度增强。拉曼特征峰在不同剂量组中的变化,为进一步从物理能级结构角度研究X射线和60Coγ射线对EC9706细胞的辐照损伤机制提供了实验依据。     

ZnO半导体电导型X射线探测器件研究

本文制备了基于ZnO纳米线阵列和ZnO薄膜的Ag-ZnO-Ag电导型X射线探测器件,研究了它们对X射线的响应特性.薄膜器件在100 V偏置时的响应度达到0.12μC/Gy,纳米线阵列器件在50 V偏压下的响应度达到0.17μC/Gy.器件工作机理研究表明,器件的响应过程与表面氧吸附与解吸附效应有关,氧气吸附与解吸附过程使得X射线辐照下的载流子寿命大幅度增加,从而使得器件对X射线具有较高的响应度.本文研究结果表明ZnO薄膜和纳米线阵列器件在X射线剂量测量领域具有应用前景.     

Characteristics of Atropa belladonna hairy roots cryopreserved by vitrification method

Atropa belladonna hairy roots (clone M8) were successfully cryopreserved by using the vitrification method. A. belladonna hairy root tips were precultured on a half strength of Murashige and Skoog (MS) solid medium with 0.1 mg per L 2,4-D or without phytohormone for 1 day, and then dehydrated with PVS2 solution for 15 minutes prior to immersion into liquid nitrogen for 1 day, 1 week, 1 month and 3 months. Hairy root tips kept in liquid nitrogen were rapidly thawed at 36 degree C in a water bath. The root tips were recultured on half strength MS medium. The hairy root tips, precultured with 2,4-D before cryopreservation, showed a higher survival rate than those precultured without phytohormone. The hairy root tips, precultured with 2,4-D, showed an average survival rate of 83 percent. There was no significant difference in the viability of the hairy roots cryopreserved for different periods. The regrowth of cryopreserved hairy roots was similar to that of untreated hairy roots and tropane alkaloid productivity became stable after 4th subculture. PCR analysis of hairy roots demonstrated the conservation of the T-DNA in cryopreserved hairy roots. These results indicate that cryopreservation by vitrification method is useful to preserve A.belladonna hairy root clone M8.     

A study on the best irradiation dose of X-ray for Hep-2 cells by Fourier transform infrared spectroscopy

Fourier transform infrared spectroscopy (FTIR) was employed to study the human epidermis larynx carcinoma cell lines (Hep-2) which were irradiated by different doses of X-ray.The results show that (1) the irradiation of X-ray damages the structure of the CH3 groups of the thymine in DNA,which restrains the reproduction of Hep-2 cells effectively,(2) the 8 Gy dose of X-ray irradiation changes the framework and the relative contents of some proteins,lipids and the nucleic acid molecules intercellular in the greatest degree,and (3) the 8 Gy dose of X-ray irradiation is the best irradiation dose for lowering the degree of the cancerization of Hep-2 cells according to the criteria for the degree of the cancerization reported recently.Meanwhile,the apoptosis of these cells were detected by using flow cytometry (FCM) primarily.It shows that the apoptotic ratio of the Hep-2 cells depends on the irradiation dose to some extent,but is not linearly.And the apoptotic ratio of the 12 Gy dose group is the maximum (20.36%),but the apoptotic ratios of the 2 to 8 Gy dose groups change little.     

12C6+离子束辐照番茄种子当代效应

利用辐射能量为80 MeV/u 的12C6+重离子束辐照番茄种子, 辐照剂量分别为30, 60, 90, 120和160 Gy研究其对番茄M1代的生物学效应。结果表明, 随着辐照剂量的增大, 番茄的发芽率和成苗率降低, 且成苗率明显低于发芽率, 发现辐照损伤主要抑制了根的生长; MDA和脯氨酸含量变化的总体趋势为随着辐照剂量的增大先升后降再升高, 说明高剂量C离子辐照对生物膜造成更严重的损伤; APX活性随着辐照剂量的增大呈先升后降再升高的趋势, 表明APX在清除活性氧中起主要作用; POD和SOD活性的总体趋势是随着辐照剂量的增大而降低, 且明显低于对照组。综合分析表明, 12C6+重离子束辐照番茄种子, 对M1代具有明显的损伤效应, 高剂量辐照对番茄种子造成的损伤更大, 使酶的活性降低, 抑制植株生长。 To investigate the M1 biological effects of heavy ions irradiation on Lycopersicon esculentum Mill., its seeds were irradiated by 12C6+heavy ions (80 MeV/u) with the dosages of 30, 60, 90, 120 and 160 Gy respectively . The results showed that with doses increased gradually, germination rate and seedling rate of Lycopersicon esculentum Mill. were decreased, and the latter was lower than the former, mainly due to the inhibition of root growth. The irradiation increased the content of MDA and proline evidently, showing irradiation could damage biomembrane, and also decreased the activities of POD and SOD with distinct inhibition pattern. However, the low dose and high dose irradiation promoted APX activity, illustrating APX was induced to protect irradiation injury. In brief, exposure to 12C6+ heavy ions had obvious injury effects on the seeds of Lycopersicon esculentum Mill.. Heavy ions irradiation damaged biomembrane, inhibited activities of enzymes, and finally inhibited the growth of the first generation of these seeds.     

重离子辐射提高腺病毒介导小鼠黑色素瘤细胞转染率的研究

探讨12C6+ 离子束辐射对用带有绿色荧光蛋白基因的缺陷性腺病毒（AdCMV GFP）转染小鼠黑色素瘤细胞(B16细胞系)的影响。 采用不同剂量的12C6+ 重离子束辐射经AdCMV GFP 转染的B16细胞， 利用流式细胞仪检测腺病毒的转染率。 结果表明， 12C6＋重离子束辐射能提高腺病毒对B16细胞的转染率， 且具有量效关系。 此外， 先转染后辐射法比起先辐射后转染法能更显著地提高转染率。The effect of 12C6+ beam irradiation on AdCMV GFP (a replication deficient recombinant adenoviral vector containing CMV promoter and green fluorescent protein) gene transfection efficiency for murine melanoma cell B16 has been investigated. B16 cells infected with AdCMV GFP were irradiated by different doses of 12C6+ beam. The transfection efficiency was assessed by flow cytometry (FCM). Results show that 12C6+ beam irradiation can improve tansfection efficiency of AdCMV GFP on murine melanoma cell B16 in a dose dependent manner. In addition， the tansfection efficiency in pre tranfection plus irradiation group is higher than that in pre irradiation plus tranfection group at the same dose irradiation dose.     

重离子束辐照对荷颊囊癌金黄地鼠血清微量元素的影响

探讨了重离子束辐照后荷颊囊癌金黄地鼠血清中微量元素含量的变化趋势。采用0, 4, 6, 8和12 Gy剂量的12C6+ 离子束对荷颊囊癌金黄地鼠辐照治疗后28 d取血, 应用原子吸收光谱仪火焰法测定血清中Fe, Cu, Zn, Mg和Ca 5种微量元素的含量。金黄地鼠成瘤后, 血清中微量Cu, Zn, Ca和Mg元素含量下降, 均明显低于正常组（P<0.05）; 不同剂量的重离子束辐照后28 d, 血清中5种微量元素在低剂量时均呈现继续下降趋势, 在6或8 Gy时恢复到正常组水平, 到12 Gy再呈降低的趋势, 存在一定的剂量-反应关系。重离子束辐照影响荷颊囊癌金地鼠血清微量元素的含量, 具有一定的临床意义。 To study the trend of the changes of trace elements in serum of golden hamster with cheek pouch carcinoma after irradiation by heavy ion beam, the cheek pouch carcinoma of golden hamster was exposed to different doses of heavy ion beam, after 4 weeks, the contents of Cu, Zn, Fe, Mg and Ca in serum were detected by flame method of Atomic Absorption Spectrometer. The contents of Cu, Zn, Ca and Mg in experimental groups with cheek pouch carcinoma were significantly lower than that of the normal group (P<0.05). After irradiated by 0, 4, 6, 8, 12 Gy heavy ion beam, the 4 Gy group showed a tendency downward, when the irradiation dose reached 6 Gy, the contents of Fe, Zn were increased, and decreased at 12 Gy. While Cu, Ca and Mg content of 8 Gy group rose to the highest, and decreased at 12 Gy. All of the results showed a dose reaction relationship (P<0.05). The irradiation of heavy ion beam maybe significantly affect the content of trace elements in serum of golden hamster with cheek pouch carcinoma.     

~(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。     

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.     

低剂量连续辐射引起的小鼠免疫系统的变化

为了评估低剂量X射线连续辐射对BALB/c小鼠健康机体免疫系统的影响, 实验采用X射线全身连续照射BALB/c小鼠, 照射第一天剂量为0.07 Gy, 剂量率0.2 Gy/min, 之后每天照射0.08 Gy, 共照射12 d, 累积剂量1.03 Gy, 照射后24和48 h取血、胸腺和脾脏。 流式细胞仪检测免疫细胞周期和凋亡的变化, 胸腺和脾脏指数用重量法获取。 实验结果表明, 小鼠胸腺细胞的周期在照射后24 h被阻滞在G2/M期; 外周血淋巴和胸腺细胞周期48 h被阻滞在 G0/G1期, 细胞凋亡比例在照射后两个时间点都显著增加; 脾脏淋巴细胞周期24 h被阻滞在 G0/G1期, 48 h被阻滞在 S期, 细胞凋亡比例在24和48 h显著减少; 脾脏指数在照射后48 h显著减少。 故低剂量X射线连续全身照射BALB/c小鼠可激活免疫细胞不同的周期监测点, 引起免疫细胞凋亡比例发生变化, 造成一定的辐射损伤, 且这种影响随着免疫器官的不同而不同。 For estimating the effect of low doses X ray continual irradiation to immunity system of mouse, BALB/c mice were continually irradiated to 1.03 Gy by X rays at a dose rate of 0.2 Gy/min in 13 d. At 24 or 48 h after irradiation, the immunocyte cycle and apoptosis were determined by flow cytometry, and the thymus and spleen weights were measured too. The results showed that the cycle of thymocyte were arrested in G2/M at 24 h, the number of peripheral blood lymphocytes and thymocytes in G0/G1 phase at 48 h was up and the percentage of apoptosis had a significance increase in both of time points; the cycle of spleen lymphocytes was delayed in G0/G1 at 24 h, in S phase at 48 h, the apoptosis had a significance decrase at 24 and 48 h; spleen index declined significantly at 48 h. The results suggested that low doses continual X ray whole body irradiation could activate different cell cycle checkpoints, and result in some changes of apoptosis and some damages to immunocytes. The continual X ray irradiation affects the organs differently, it might provide experiment basis for radioprotection.