~(60)Coγ射线辐射对淋巴细胞化学发光的生物效应(英文)

用淋巴细胞化学发光(Ly-CL)技术,研究了离体状况下~(60)Coγ射线辐射对PHA或ConA激发的Ly-CL的生物效应。结果表明:(1)0.25Gy~(60)Coγ射线辐射对PHA(200μg/mL)激发的Ly-CL峰值有增强趋势(P>0.05),0.5Gy有明显增强效应(P<0.05),1～16Gy引起显著或非常显著性抑制(P<0.05或P<0.01)。(2)2～16Gy~(60)Coγ射线辐射对ConA(50、100μg/mL)激发的Ly-CL峰值产生显著或非常显著性抑制作用(P<0.05或 P<0.01)。回归分析表明,峰值下降幅度和辐射剂量间有显著线性关系(P<0.0005)。2～16Gy~(60)Coγ射线辐射对PHA或ConA引起的Ly-CL峰时无影响(P>0.05)。     

12C6+重离子辐照对人肝细胞、肝癌细胞端粒酶活性表达的影响

为探索碳离子束辐照对细胞中端粒酶活性的变化,利用兰州近代物理研究所重离子研究装置(Heavy ionresearch facility in lanzhou,HIRFL)产生的碳离子(31MeV/μ12C6 ),以人肝细胞HL-7702,肝癌细胞SMMC-7721为实验对象,用不同剂量1Gy、2Gy、3Gy、4Gy的重离子分别对两种细胞进行照射,用多聚酶链式反应.银染端粒重复序列扩增法(PCR-telomeric repeat amplification protocol,TRAP-PCR)银染端粒重复序列扩增法检测不同剂量下细胞端粒酶活性的变化.结果显示,人肝细胞HL-7702自身没有端粒酶活性,经1Gy辐照后也没有端粒酶活性,在2和3Gy处出现端粒酶活性,4Gy处端粒酶活性又消失.肝癌细胞SMMC-7721在1～3Gy处随着剂量的增大端粒酶活性升高,在4Gy处又开始下降;在1～3Gy处随着时间的推移端粒酶活性随着时间而加强(p<0.05).分析得知,重离子辐射可以诱导人肝细胞产生端粒酶活性,也可以改变肝癌细胞的端粒酶活性.端粒酶参与细胞受辐照后DNA单链损伤的修复;辐照后DNA双链断裂导致端粒酶活性减弱.本实验使重离子在辐照治疗中的优势得以体现.     

~(60)Coγ射线对小鼠巨噬细胞和淋巴细胞胞浆微管的影响

本实验采用间接免疫荧光技术,观察了小鼠腹腔巨噬细胞和肠系膜淋巴结细胞胞浆微管经~(60)Coγ射线照射后的变化及分布特点;同时还对此变化进行了动态观察、研究;并在透射及扫描电镜下观察了~(60)Coγ射线对胞浆内微管、中心粒和表面微绒毛的影响。实验结果表明:(1)辐照对巨噬、淋巴细胞内胞浆微管的分布及荧光染色模式有明显影响,并且与辐照剂量有关;(2)经一定时间后,由辐照所致的胞浆微管变化可以完全恢复;(3)透射电镜下可见辐照后胞浆微管、中心粒基本消逝,扫描电镜下可见经辐照后细胞表面突起或微绒毛发生明显变化。     

美洲商陆丝裂原(PWM)诱导淋巴细胞和脂多糖(LPS)诱导淋巴细胞的辐射敏感性

应用人血在体外培养,分别用PWM和LPS诱导淋巴细胞,接受各种剂量~(60)Coγ线照射,两种细胞按各种组合混合再培养,并设有各种单项培养作对照,以~3H-TdR掺入反映淋巴细胞的增殖转化。实验结果说明PWM细胞能够激活LPS细胞转化,当一种细胞受照后单独培养或与另一种正常细胞混合培养,其放射性掺入即下降,并随剂量增加而愈益明显、下降趋势均呈现负相关,并分别得到直线回归方程,差异显著性比较说明PWM细胞受照影响更严重。当PWM细胞接受1Gy以上剂量后与正常LPS细胞混合培养即失去激活作用,LPS细胞受照1—2 Gy仍然可被激活,以上提示电离辐射导致机体免疫缺陷,其中PWM诱导的T辅助细胞受损是较重要的。     

~(60)Co γ射线对小鼠巨噬细胞和淋巴细胞浆微管的影响

本实验采用间接免疫荧光技术,观察了小鼠腹腔巨噬细胞和肠系膜淋巴结细胞胞浆微管经(60)Coγ射线照射后的变化及分布特点;同时还对此变化进行了动态观察、研究;并在透射及扫描电镜下观察了(60)Coγ射线对胞浆内微管、中心粒和表面微绒毛的影响。实验结果表明:(1)辐照对巨噬、淋巴细胞内胞浆微管的分布及荧光染色模式有明显影响,并且与辐照剂量有关;(2)经一定时间后,由辐照所致的胞浆微管变化可以完全恢复;(3)透射电镜下可见辐照后胞浆微管、中心粒基本消逝,扫描电镜下可见经辐照后细胞表面突起或微绒毛发生明显变化。     

~(60)Co-γ线对刀豆球蛋白(ConA)和脂多糖(LPS)诱导的淋巴细胞的效应

应用~3H-TdR掺入法研究ConA和LPS诱导后的淋巴细胞亚群的辐射效应及其相互关系。体外实验说明ConA细胞具有激活LPS对B细胞的诱导作用。当ConA细胞受10Gy照射后~3H-TdR掺入明显下降,并失去激活作用。LPS细胞受照不能被正常ConA细胞所激活。两种细胞受照后混合培养出现ConA细胞对LPS细胞的抑制作用。应用琼脂扩散盒培养法反映辐射对两种细胞的效应更明显。 8例鼻咽癌病人接受一个疗程的~(60)Coγ线治疗后,其ConA细胞的掺入显著下降,失去激活B细胞的作用。LPS细胞不能被正常ConA细胞所激活。本实验提示ConA细胞的放射敏感性比LPS细胞高。     

~(12)C离子照射对MDM2和GADD45A基因表达的影响

12C离子照射人淋巴细胞株Peng-EBV,剂量率约为0.3~0.5 Gy/min,辐照剂量为0、0.1、0.3、0.5、0.7、1.0、2.0 Gy。采用荧光定量PCR技术检测不同剂量点、不同时间点的MDM2和GADD45A基因相对表达水平。结果表明,人淋巴细胞株受照后,MDM2和GADD45A基因相对表达水平整体升高;MDM2基因在0.5~2.0 Gy、照射后2和12小时,以及GADD45A基因在0.7~2.0 Gy、照射后2、12、24和48小时,相对表达水平均有随着照射剂量的增加而上升的趋势。     

低剂量12C6+离子辐照对小鼠胸腺、脾脏细胞周期及DNA损伤的影响

研究低剂量12C6+子全身辐照对小鼠胸腺、脾脏细胞周期进程及DNA损伤的影响.以0、10、50、75、100和250mGy 12C6+离子全身辐照小鼠,照射后6h处死小鼠,用流式细胞仪检测受辐照小鼠胸腺、脾脏细胞在各细胞周期的百分率,用彗星电泳技术检测受辐照小鼠胸腺脾脏细胞的拖尾率和拖尾长度.所有照射组G0/G1期胸腺细胞百分率明显低于对照组,(p<0.05),10～100mGy照射组S期胸腺细胞百分率显著高于对照组(p<0.01),所有照射组(G2/M)期胸腺细胞百分率明显高于对照组(p<0.05);所有照射组G0/G1期脾细胞百分率明显高于对照组(p<0.01),S期脾细胞百分率显著低于对照组(p<0.05).彗星电泳结果显示低剂量12C6+离子辐照以剂量依赖的方式引起小鼠胸腺脾脏细胞DNA迁移长度及拖尾率的增加.低剂量的碳离子辐射可促进小鼠胸腺细胞DNA合成,对小鼠脾脏细胞产生抑制作用,使其发生G1期阻滞;同时对胸腺及脾脏细胞造成具有明显剂量效应关系的DNA损伤.     

30MeV/u~(40) Ar~(15+) 和 ~(60)Co γ 射线对人肝癌细胞株 SMMC-7721 辐射效应的比较

比较了重离子３０ＭｅＶ／ｕ４０Ａｒ１５＋和６０Ｃｏγ射线对体外培养的人肝癌细胞株ＳＭＭＣ７７２１的辐射效应，用活体和固定的方法观察了辐射处理后细胞死亡的过程。结果表明：无论是重离子还是γ射线，都能对细胞造成严重损伤，影响细胞存活百分数；同时，重离子比γ射线对细胞有更显著的杀伤效果，在处理后第４８ｈ，细胞存活率为５０％时，重离子的ＲＢＥ约为１５。     

低剂量~(60)Coγ线照射对人淋巴细胞质膜的效应

本文报道了低剂量~(60)Coγ线照射后人外周血淋巴细胞质膜表面受体功能和结构的变化。结果表明,25拉德照射后淋巴细胞活性 E 玫瑰(E_a)花环形成百分率显著减少(p<0.01),100拉德照射后未见进一步减少;25拉德照射后淋巴细胞小鼠红细胞玫瑰(M_E)花环形成百分率明显下降(p<0.01),100拉德照射后下降更显著;25拉德照射后淋巴细胞膜表面免疫球蛋白 G 阳性(S_mI_gG~+)细胞百分率减少,100拉德照射后减少更显著(p<0.01);100拉德照射后由电镜可见淋巴细胞质膜膨出,细胞核未见异常。     

Evaluation of sex specificity on oxidative stress induced in lungs of mice irradiated by 12C6+ions

The aim of this work is to identify if there is sex specificity on 12C6 ion-induced oxidative damage in mouse lung at different time points. Kun-Ming mice were divided into two groups, each composed of six males and six females: control group and irradiation group with a single acute dose of 4 Gy. Animals were sacrificed at 2, 4 and 12 h respectively, there lungs were removed immediately, and the oxidative stress-related biomarkers were measured by Diagnostic Reagent Kits. The results showed that the relative activities of superoxide dismutase (4 h), catalase (2 h) and Se-dependent glutathione peroxidase (12 h) have significant changes (P<0.05) between male groups and female groups, suggesting that the lungs of male mice are more sensitive to counteracting the oxidative challenge. Moreover, higher levels of malondiadehyde and lower contents of glutathione were also found in males, indicating that oxidative stress induced by 12C6 ion is pronounced in the lungs of males. We thought that these sex-responded differences may be attributed to the influence of sex hormones.     

Assessment of biological changes in wheat seedlings induced by ^12C^6＋-ion irradiation

The present study was designed to evaluate the effect of 12C6 ion beam (10～80 Gy) on biological changes of wheat seedlings. Reactive oxygen species (ROS)-related biomarkers and the quantification of plant survival and growth were examined at 10 day after carbon ions irradiation (LET 30.8keV/μm). The results showed that heavy ions obviously enhanced ROSs reflected by the production of O2 and H2O2 as well as TBARS, and treatment with 20 Gy achieved the peak value, suggesting that higher mutagenic potential may occur at 20 Gy. Simultaneously, increase of SOD activity was induced by heavy ions to counteract ROS accumulation. On the other hand, higher doses at 40 and 80 Gy inhibited wheat growth and survival in comparison with the control, and reversely lower doses at 10 or 20 Gy stimulated wheat growth and survival. In conclusion, the above observations imply that a dose range of 20～40 Gy is likely promised for wheat mutation breeding.     

A high-throughput screening method for breeding Aspergillus niger with 12C6+ ion beam-improved cellulase

In this study,a high-throughput screening method was established through the 24-square deep-well microliter plate(MTP) fermentation and micro-plate detection for large-scale screening of the mutants.It was suitable for screening a large number of mutants and improving the breeding efficiency after heavy-ion beam irradiation.Seventeen strains showed higher cellulase activity compared with the initial strain after the screening of plate and MTP fermentation.The filter paper activity and β-glucosidase activity of Aspergillus niger H11201 had increased 38.74 and 63.23%separately compared with A.niger H11 by shaking flask fermentation,and it was genetically stable after being passaged to nine generations.The results indicate that the high-throughput screening method can be used for the quick breeding of A.niger with high cellulase activity.     

Antitumor and radiosensitization effect of 12C6+ heavy-ion irradiation mediated by radiation-inducible gene therapy

Radio genetic therapy which combines gene therapy with radiotherapy has shown promising results in cancer treatment. In this study, an oncolytic adenovirusbased gene therapy system regulated by radiation was constructed to improve the cancer curative effect. This gene therapy system incorporated the radiation-inducible early growth response gene(Egr-1) promoter and the anticancer gene tumor necrosis factor-related apoptosis-inducing ligand(TRAIL). To confirm the antitumor effect of Ad-ET combined with~12C~(6+)tion irradiation, the survival and apoptosis fraction of tumor cells HT1080 and normal cells MRC-5 in combination treatment were detected by CCK-8 assay and FACS analysis. Then the expression levels of TRAIL gene and protein were tested by real-time PCR and western blotting. The results show that~12C~(6+)tion irradiation could induce cell growth inhibition and apoptosis by activating the TRAIL gene expression in tumor cells, while exhibiting no obvious toxicity to the normal lung cell line MRC-5. Theresults also demonstrate that use of an oncolytic adenovirusbased radiation-inducible gene therapy system together with~12C~(6+)tion irradiation could cause synergistic antitumor effect specifically in tumor cells but not in normal cells. The results indicate that the novel radio genetic therapy could potentiate radiation treatment by improving the safety and efficiency of monotherapy, and provide theoretical support for clinical application of combination treatment.     

当归红芪超滤物对重离子12C6+辐射肝癌细胞DNA损伤修复的影响

将人肝癌H22细胞分成4组,分别为对照组、药物组(100 mg/L)、辐射组(2 Gy)及联合组(100 mg/L药物 + 2 Gy照射),采用CCK-8法、单细胞凝胶电泳、γ-H2AX免疫荧光原位杂交技术以及Western Blotting印迹法,研究当归红芪超滤物(Radix Angelicae Sinensis and Radix Hedysari, RAS-RH)对重离子¹²C⁶⁺辐射引起人肝癌H22细胞DNA损伤修复的影响和其可能的机制。结果表明,在0～72 h和给药剂量为5～200 mg/L范围内,RAS-RH对人肝癌H22细胞的增殖抑制作用具有时间和剂量依赖性,其20%抑制浓度IC₂₀为(117.6±2.15)mg/L;单细胞凝胶电泳显示联合组头部DNA含量低于辐射组,而尾部DNA含量、尾距TM、Olive尾距OTM均高于辐射组;γ-H2AX免疫荧光原位杂交技术发现RAS-RH不增加重离子¹²C⁶⁺辐射引起的DNA损伤,但在2—12 h,DNA双链断裂的γ-H2AX foci修复作用被RAS-RH抑制,DNA损伤持续存在;Western Blotting显示RAS-RH通过下调Ku70/80及Rad51的蛋白表达,抑制γ-H2AX的聚集。以上结果说明RAS-RH对人肝癌H22细胞的辐射增敏作用可能是下调DNA损伤修复相关因子Ku70/80及Rad51的表达。     

Evaluation of sex specificity on oxidative stress induced in lungs of mice irradiated by ^12C^6＋ ions

The aim of this work is to identify if there is sex specificity on ^12C^6＋ ion-induced oxidative damage in mouse lung at different time points. Kun-Ming mice were divided into two groups, each composed of six males and six females： control group and irradiation group with a single acute dose of 4 Gy. Animals were sacrificed at 2, 4 and 12 h respectively, there lungs were removed immediately, and the oxidative stress-related biomarkers were measured by Diagnostic Reagent Kits. The results showed that the relative activities of superoxide dismutase （4 h）, catalase （2 h） and Se-dependent glutathione peroxidase （12 h） have significant changes （P〈0.05） between male groups and female groups, suggesting that the lungs of male mice are more sensitive to counteracting the oxidative challenge. Moreover, higher levels of malondiadehyde and lower contents of glutathione were also found in males, indicating that oxidative stress induced by ^12C^6＋ ion is pronounced in the lungs of males. We thought that these sex-responded differences may be attributed to the influence of sex hormones.