辐照诱导的人正常肝细胞系HL-7702细胞远后效应

近年来，放射治疗在肿瘤治疗中的作用受到了越来越多的重视，放疗技术和手段的迅速发展为人们选择放疗创造了更多的机会[1]。放疗是利用电离辐射对细胞，特别是细胞中的遗传物质DNA的损伤作用，诱发病灶部位不正常细胞的凋亡和坏死来治疗肿瘤的。即使制定了周密的放疗方案和计划，放疗过程中难免也会对正常组织和细胞造成一定程度的损伤，而且辐射的远后效应和旁效应的发现，也使得放射治疗肿瘤的安全性受到了很大的关注。要想充分发挥放疗的优势，而又使其对人体正常组织的毒副作用尽可能降低，就必须搞清楚电离辐射对细胞造成的各种损伤的类型及其分子机理。自从电离辐射的远后效应和旁效应被发现以来就一直是辐射生物医学领域的研究热点，国内外的科研人员进行了大量的实验试图解释它们的本质，可是几十年来相关领域的研究一直局限于细胞学的水平，在向分子水平前进的道路上遇到了巨大的困难[2,3]。目前对于电离辐射远后效应和旁效应的研究是相对独立进行的，很少有探讨二者相互关系的论文发表。本文试图在总结前人科研成果并结合对自己所取得的实验数据进行分析的基础上对电离辐射的远后效应、旁效应、基因不稳定性与电离辐射所产生的活性氧自由基的关系进行初步的探讨。实验方法与结果： 1.用X射线辐照人正常肝细胞系HL-7702细胞，运用胞质分离阻滞微核实验检测细胞微核率，AnnexinV-FITC细胞凋亡检测试剂盒检测细胞凋亡率，细胞微核率和凋亡率随着辐照剂量的增加而显著增加。X射线照射后细胞继续传代培养，第七代时不同剂量辐照后子代细胞微核率和凋亡率同未辐照细胞的微核率和凋亡率相比已经没有明显区别。对不同剂量辐照后传代七代的细胞再次照射2.5Gy相同的剂量，发现受初次不同剂量辐照的细胞其微核率和凋亡率再次出现明显差异，初次辐照剂量高的细胞再次相同剂量辐照后的微核率和凋亡率也高。 2.对不同剂量X射线辐照后的细胞继续传代到第十五代时用H2O2浓度为1ul/ml的培养基处理15min，发现受初次不同剂量辐照的细胞其微核率再次出现比较明显差异，初次辐照剂量高的细胞H2O2处理后的微核率也高。 3.对受到不同剂量X射线辐照的人正常肝细胞存活后代进行二次辐照，分两组分别给予1.5GyX射线和1Gy碳离子束辐照，并检测二次辐照后细胞的微核率，发现重离子束二次辐照后并不像X射线一样可以诱发初次X射线辐照造成的损伤信息的表达，而只是表现出二次重离子辐照所造成的损伤。结论： 1.X射线辐照导致了HL-7702细胞基因组不稳定性这一辐射远后效应，X射线二次辐照辐照存活细胞的子代细胞可以诱发辐射的远后效应（如基因组不稳定性）明显的表现； 2.X射线辐照导致的HL-7702细胞后代基因组不稳定性，可以通过H2O2处理而得以诱发，揭示电离辐射过程中产生的氧自由基可能与辐射的远后效应存在密切的联系； 3.电离辐射产生的ROS在诱发细胞产生微核中具有重要的作用，但是并非唯一的影响因素。 4.X射线和12C6+重离子束辐照后存活细胞的后代中的细胞损伤类型可能存在着本质上的区别

Antioxidant activity and cytoprotective effect of kappa-carrageenan oligosaccharides and their different derivatives

Antioxidant activity of kappa-carrageenan oligosaccharides (OM) and their chemical modification derivatives was investigated employing various established in vitro systems, such as reducing power, iron ion chelation, and total antioxidant activity using beta-carotene-linoleic acid system. The oversulfated (SD), lowly (LAD), and highly acetylated derivatives (HAD) in reducing power assay, the phosphorylated derivative (PD) in metal chelating assay, and oversulfated and phosphorylated derivatives in total antioxidant activity assay exhibited antioxidant activity higher than that of carrageenan oligosaccharides. The results indicated that the chemical modification of carrageenan oligosaccharides can enhance their antioxidant activity in vitro. The protective effects of the carrageenan oligosaccharides and their chemically modified derivatives against H2O2 and UVA (long-wave ultraviolet radiation) induced oxidative damage on rat thymic lymphocyte were investigated by measuring cell viability via 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT). Thymic lymphocyte exposure to H2O2 and UVA, a marked reduction in cell survival was observed, which was significantly prevented by carrageenan oligosaccharides and their derivatives (preincubated for 2 h) at 66.7-2000 mu g/mL. But both the carrageenan oligosaccharides and their different derivatives showed the similar protective effects on intracellular level. Taken together, these results suggest that carrageenan oligosaccharides and their derivatives show relevant antioxidant activity both in vitro and in a cell system. (C) 2005 Elsevier Ltd. All rights reserved.

High-efficiency transfer and expression of AdCMV-p53 in human cervix adenocarcinoma cells induced by subclinical-dose carbon beam radiation

Purpose The aim of this study is to evaluate the eVect of carbon-beam irradiation on adenovirus-mediated p53 transfer in human cervix adenocarcinoma.Materials and methods The HeLa cells pre-exposed to carbon-beam or -ray, were infected with replication-deficient adenovirus recombinant vectors, containing human wild-type p53 (AdCMV-p53) and green Xuorescent protein (GFP) (AdCMV–GFP), respectively. The GFP transfer and p53 expression were detected by Xow cytometric analysis.Results The GFP transfer frequency in C-beam with AdCMV-GFP groups was 38–50% more than that inγ-ray with AdCMV–GFP groups. The percentage of p53 positive cells in the C-beam with AdCMV–p53 groups was 34–55.6% more than that in γ-ray with AdCMV-p53 groups (p < 0.05), suggesting that subclinical-dose C-beam irradiation could signiWcantly promote exogenous p53 transfer and p53 expression, and extend the duration of p53 expression in the HeLa cells. The expression of p21 increased with p53 expression in HeLa cells. The survival fractions for the 0.5–1.0 Gy C-beam with AdCMV-p53 groups were 38–43% less than those for the isodose γ-ray with AdCMV-p53 groups, and 31–40% less than those for the C-beam only groups (p <0.05).Conclusions The subclinical-dose C-beam irradiation could signiWcantly promote the transfer and expression of exogenous p53, extend the duration of p53 expression, and enhance the suppression of p53 on cervix adenocarcinoma cells.

High-efficiency transfer and expression of AdCMV-p53 in human hepatocellular carcinoma cells induced by low-dose carbon-ion radiation

Objective To investigate whether the irradiation with C-beam could enhance adenovirus-mediated transfer and expression of p53 in human hepatocellular carcinoma. Materials and methods HepG2 cells were exposed to C-beam or gamma-ray and then infected with replicationdeficient adenovirus recombinant vectors containing human wild-type p53 or green fluorescent protein, respectively. The transfer efficiency and expression level of the exogenous gene were detected by flow cytometric analysis. Cell survival fraction was detected by clonogenic assay. Results The transfer frequency in C-beam or gamma-irradiated groups increased by 50-83% and 5.7-38.0% compared with the control, respectively (P < 0.05). Compared with C-beam alone, p53 alone, and gamma-ray with p53, the percentages of p53 positive cells for 1 Gy C-beam with p53 increased by 56.0-72.0%, 63.5-82.0%, and 31.3-72.5% on first and third day after the treatments, respectively (P < 0.05). The survival fractions for the 2Gy C-bearn and AdCMV-p53 infection groups decreased to similar to 2%. Conclusion C-beam irradiation could significantly promote AdCMV-green fluorescent protein transfer and expression of p53.

Survivin表达在高LET射线诱导细胞凋亡中作用机理的初步研究；A Preliminary Study on Action Mechanisms of Survivin Expression in Cell Apoptosis Induced by High-LET Radiation

先前的研究表明,肿瘤细胞中survivin的高表达与细胞对高传能线密度(LET)射线的辐射抗性相关。研究了survivin表达在高LET射线诱导的细胞凋亡中的作用,发现抑制survivin表达对高LETC离子辐射诱导的Bcl-2和Bax表达没有明显的影响。在高LET射线辐照中,survivin可能通过抑制caspase-3和-9活性的途径,抑制了细胞凋亡。

Vulcanization of polybutadiene latex induced by Co-60 gamma radiation

Polybutadiene latex (PBL) vulcanization induced by Co-60 radiation and the influence of dose on crosslinking were investigated. Morphology and particle size distribution were examined by AFM and a particle size analyzer. The casting films were characterized for their swelling and mechanical properties as a function of dose.

Production of active oxygen species from Taxus cuspidata induced by far-UV radiation

In order to understand the role of active oxygen species in mediating plant injuries induced by far-UV radiation, seedlings of Taxus cuspidata Sieb. et Zucc. were irradiated by far-UV rays in laboratory for 4 weeks. The production of organic free-radicals in detached needles, and the production of O-2(radical anion) and O-1(2) in isolated chloroplasts were detected weekly by electron spin resonance (ESR) to evaluate their relative importance. The results show that the cumulative effect of far-UV irradiation, is best indicated by the production of organic free radicals in the needles, O-2(radical anion) production in chloroplasts is the next. The enhancement of O-1(2) production in chloroplasts by the cumulative far-UV irradiation seems to be not so important as O-2(radical anion) in mediating injuries induced by, far-UV radiation because of its high background value.

Strain H-2-419-4 of Haematococcus pluvialis induced by ethyl methanesulphonate and ultraviolet radiation

Two strains H-2-410 and H-2-419 were obtained from the chemically mutated survivors of wild Haematococcus pluvialis 2 by using ethyl methanesulphonate (EMS). Strains H2-410 and H2-419 showed a fast cell growth with 13% and 20% increase in biomass compared to wild type, respectively. Then H-2-419-4, a fast cell growth and high astaxanthin accumulation strain, was obtained by exposing the strain H2-419 to ultraviolet radiation (UV) further. The total biomass, the astaxanthin content per cell, astaxanthin production of H-2-4194 showed 68%, 28%, and 120% increase compared to wild H. pluvialis 2, respectively. HPLC (High Performance Liquid Chromatography) data showed also an obvious proportional variation of different carotenoid compositions in the extracts of H2-4194 and the wild type, although no peak of carotenoids appeared or disappeared. Therefore, the main compositions in strain H-2-419-4, like its wild one, were free of astaxanthin, monoester, and diester of astaxanthin. The asexual reproduction in survivors after exposed to UV was not synchronous, and different from the normal synchronous asexual reproduction as the mother cells were motile instead of non-motile. Interestingly, some survivors from UV irradiation produced many mini-spores (or gamete?), the spores moved away from the mother cell gradually 4 or 5 days later. This is quite similar to sexual reproduction described by Elliot in 1934. However, whether this was sexual reproduction remains questionable, as no mating process has been observed.

Gamma-ray induced color centers in Yb : YAG crystals grown by Czochralski method

20 at.% Yb:YAG single crystals have been grown by the CZ method and gamma-ray irradiation induced color centers and valence change of Fe3+ and Yb3+ ions in Yb:YAG have been studied. One significant 255 nm absorption band was observed in as-grown crystals and was attributed to Fe3+ ions. Two additional absorption (AA) bands located at 255 nm and 345 nm, respectively, were produced after gamma irradiation. The changes in the AA spectra after gamma irradiation and air annealing are mainly related to the charge exchange of the Fe3+, Fe2+, oxygen vacancies and F-type color centers. Analysis shows that the broad AA band is associated with Fe2+ ions and F-type color centers. The transition Yb3+ Yb2+ takes place as an effect of recharging of one of the Yb3+ ions from a pair in the process of gamma irradiation. (C) 2006 Elsevier Ltd. All rights reserved.

UV-B-induced Oxidative Damage and Protective Role of Exopolysaccharides in Desert Cyanobacterium Microcoleus vaginatus

UV-B-induced oxidative damage and the protective effect of exopolysaccharides (EPS) in Microcoleus vaginatus, a cyanobacterium isolated from desert crust, were investigated. After being irradiated with UV-B radiation, photosynthetic activity (Fv/Fm), cellular total carbohydrates, EPS and sucrose production of irradiated cells decreased, while reducing sugars, reactive oxygen species (ROS) generation, malondialdehyde (MDA) production and DNA strand breaks increased significantly. However, when pretreated with 100 mg/L exogenous EPS, EPS production in the culture medium of UV-B stressed cells decreased significantly; Fv/Fm, cellular total carbohydrates, reducing sugars and sucrose synthase (SS) activity of irradiated cells increased significantly, while ROS generation, MDA production and DNA strand breaks of irradiated cells decreased significantly. The results suggested that EPS exhibited a significant protective effect on DNA strand breaks and lipid peroxidation by effectively eliminating ROS induced by UV-B radiation in M. vaginatus.

Light histories influence the impacts of solar ultraviolet radiation on photosynthesis and growth in a marine diatom, Skeletonema costatum

Phytoplanktonic species acclimated to high light are known to show less photoinhibition. However, little has been documented on how cells grown under indoor conditions for decades without exposure to UV radiation (UVR, 280-400 nm) would respond differently to solar UVR compared to those in situ grown under natural solar radiation. Here, we have shown the comparative photosynthetic and growth responses to solar UVR in an indoor-(IS) and a naturally grown (WS) Skeletonema costatum type. In short-term experiment (<1 day), phi(PSII) and photosynthetic carbon fixation rate were more inhibited by UVR in the IS than in the WS cells. The rate of UVR-induced damages of PSII was faster and their repair was significantly slower in IS than in WS. Even under changing solar radiation simulated for vertical mixing, solar UVR-induced higher inhibition of photosynthetic rate in IS than in WS cells. During long-term (10 days) exposures to solar radiation, the specific growth rate was much lower in IS than WS at the beginning, then increased 3 days later to reach an equivalent level as that of WS. UVR-induced inhibition of photosynthetic carbon fixation in the IS was identical with that of WS at the end of the long-term exposure. The photosynthetic acclimation was not accompanied with increased contents of UV-absorbing compounds, indicating that repair processes for UVR-induced damages must have been accelerated or upgraded. (C) 2008 Elsevier B.V. All rights reserved.

Combined effects of ultraviolet radiation and temperature on morphology, photosynthesis, and DNA of Arthrospira (Spirulina) platensis (Cyanophyta)

Natural levels of solar UVR were shown to break and alter the spiral structure of Arthrospira (Spirulina) platensis (Nordst.) Gomont during winter. However, this phenomenon was not observed during summer at temperatures of similar to 30 degrees C. Since little has been documented on the interactive effects of solar UV radiation (UVR; 280-400 nm) and temperature on cyanobacteria, the morphology, photosynthesis, and DNA damage of A. platensis were examined using two radiation treatments (PAR [400-700 nm] and PAB [PAR + UV-A + UV-B: 280-700]), three temperatures (15, 22, and 30 degrees C), and three biomass concentrations (100, 160, and 240 mg dwt [dry weight] . L-1). UVR caused a breakage of the spiral structure at 15 degrees C and 22 degrees C, but not at 30 degrees C. High PAR levels also induced a significant breakage at 15 degrees C and 22 degrees C, but only at low biomass densities, and to lesser extent when compared with the PAB treatment. A. platensis was able to alter its spiral structure by increasing helix tightness at the highest temperature tested. The photochemical efficiency was depressed to undetectable levels at 15 degrees C but was relatively high at 30 degrees C even under the treatment with UVR in 8 h. At 30 degrees C, UVR led to 93%-97% less DNA damage when compared with 15 degrees C after 8 h of exposure. UV-absorbing compounds were determined as negligible at all light and temperature combinations. The possible mechanisms for the temperature-dependent effects of UVR on this organism are discussed in this paper.

Vertical mixing within the epilimnion modulates UVR-induced photoinhibition in tropical freshwater phytoplankton from southern China

1. The importance of vertical mixing in modulating the impact of UVR on phytoplankton photosynthesis was assessed in a tropical, shallow lake in southern China from late winter to mid-spring of 2005. 2. Daily cycles of fluorescence measurements (i.e. photosynthetic quantum yield, Y) were performed on both 'static' and in situ samples. Static samples were of surface water incubated at the surface of the lake under three radiation treatments - PAB (PAR + UVR, 280-700 nm), PA (PAR + UV-A, 320-700 nm) and P (PAR, 400-700 nm). In situ samples were collected every hour at three different depths - 0, 0.5 and 1 m. 3. The general daily pattern was of a significant decrease in Y from early morning towards noon, with partial recovery in the afternoon. Samples incubated under static conditions always had lower Y than those under in situ conditions at the same time of the day. 4. Under stratified conditions, no overall impact of UVR impact could be detected in situ when compared with the static samples. Further rapid vertical mixing not only counteracted the impact of UVR but also stimulated photosynthetic efficiency. 5. Based on these measurements of fluorescence, the mixing speed of cells moving within the epilimnion was estimated to range between 0.53 and 6.5 cm min(-1). 6. These data show that mixing is very important in modulating the photosynthetic response of phytoplankton exposed to natural radiation and, hence, strongly conditions the overall impact of UVR on aquatic ecosystems.

Solar UV radiation drives CO2 fixation in marine phytoplankton: A double-edged sword

Photosynthesis by phytoplankton cells in aquatic environments contributes to more than 40% of the global primary production (Behrenfeld et al., 2006). Within the euphotic zone (down to 1% of surface photosynthetically active radiation [PAR]), cells are exposed not only to PAR (400-700 nm) but also to UV radiation (UVR; 280-400 nm) that can penetrate to considerable depths (Hargreaves, 2003). In contrast to PAR, which is energizing to photosynthesis, UVR is usually regarded as a stressor (Hader, 2003) and suggested to affect CO2-concentrating mechanisms in phytoplankton (Beardall et al., 2002). Solar UVR is known to reduce photosynthetic rates (Steemann Nielsen, 1964; Helbling et al., 2003), and damage cellular components such as D1 proteins (Sass et al., 1997) and DNA molecules (Buma et al., 2003). It can also decrease the growth (Villafane et al., 2003) and alter the rate of nutrient uptake (Fauchot et al., 2000) and the fatty acid composition (Goes et al., 1994) of phytoplankton. Recently, it has been found that natural levels of UVR can alter the morphology of the cyanobacterium Arthrospira (Spirulina) platensis (Wu et al., 2005b). On the other hand, positive effects of UVR, especially of UV- A (315-400 nm), have also been reported. UV- A enhances carbon fixation of phytoplankton under reduced (Nilawati et al., 1997; Barbieri et al., 2002) or fast-fluctuating (Helbling et al., 2003) solar irradiance and allows photorepair of UV- B-induced DNA damage (Buma et al., 2003). Furthermore, the presence of UV-A resulted in higher biomass production of A. platensis as compared to that under PAR alone (Wu et al., 2005a). Energy of UVR absorbed by the diatom Pseudo-nitzschia multiseries was found to cause fluorescence (Orellana et al., 2004). In addition, fluorescent pigments in corals and their algal symbiont are known to absorb UVR and play positive roles for the symbiotic photosynthesis and photoprotection (Schlichter et al., 1986; Salih et al., 2000). However, despite the positive effects that solar UVR may have on aquatic photosynthetic organisms, there is no direct evidence to what extent and howUVR per se is utilized by phytoplankton. In addition, estimations of aquatic biological production have been carried out in incubations considering only PAR (i. e. using UV-opaque vials made of glass or polycarbonate; Donk et al., 2001) without UVR being considered (Hein and Sand-Jensen, 1997; Schippers and Lurling, 2004). Here, we have found that UVR can act as an additional source of energy for photosynthesis in tropical marine phytoplankton, though it occasionally causes photoinhibition at high PAR levels. While UVR is usually thought of as damaging, our results indicate that UVR can enhance primary production of phytoplankton. Therefore, oceanic carbon fixation estimates may be underestimated by a large percentage if UVR is not taken into account.