Survey of biosystematic and sexual reproductive physiology in Holothuroidea on Hormozgan Province

Sea cucumbers belong to phylum Echinodermata, order Holothuroidea are an abundant and diverse group of Invertebrates, with over 1400 species occuring from the intertidal to the deepest oceanic trenches. Sea cucumbers are important components of the food chain in temperate and coral reef ecosystems and they play an important role as deposite feeders and suspension feeders. Rapid decline in populations may have serious consequences for the survival of other species that are part of the same complex food web,as the eggs, larve and juveniles constitute an important food source for the other marine species including crustaceans, fish and mollusks. In addition sea cucumbers are often called the earthworms of the sea, because they are responsible for the extensive shifting and mixing of the substrate, and recycling of detrital matter. Sea cucumbers consume and grind sediment and organic material into finer particles , turning over the top layers of sediment in lagoons , reefs and other habitats and allowing the penetration of oxygen. While the taxonomy of the holothurian families is generally well known , the distinction of similar species is difficult. There are relatively few holothurian taxonomist.Most sea cucumber species can be identified by Holothurin taxonomists by using the calcareous skeletal ossicles found in the body wall. In this study , at first a sea cucumber from Kish island in Persian gulf has recognized. Individuals collected from west and east extend far away into north and south of coral reefs by diving. I have checked them morphologically and anatomically.Then with key to the orders of the Holothuroidea, They belong to the Aspidochirotida with key to the families of Aspidochirotida, they were in Stichopodidae families and with key to the genus of Stchopodidae, they were Stichopus. Then ossicles were extracted at National Museum of Natural History, by Dr David Pawson. The ossicles were measured on a transect across a slide prepared from the mid-dorsal region of each specimen.The one we have in the shallow waters of Kish island, is Stichopus hermanni, a massive holothurian, body broad, considerably flattened ventraly ,the dorsal side slightly arched and the lateral sides almost vertical; body wall fairy thick and soft ; mouth subterminal; anus central; tentacles usually 20 in number of length and leaf shaped. Numerous ossicles consisting of table with large discs having usually 7 to 15 peripheral holes, but often irregular or incomplete and spire of moderate height ending in a group of spinelets, rosettes of variable development, and c-shaped rods. Color (exept papillae)partly remained after preservation in alcohol which is found at the depth of 4 to 8 meters, on coral reef. Furthermore, the sexual reproductive cycle was described using standard methods. Gonads were removed and transferred to Bouin's fixative for four weeks and then processed according to standard embedding technique. To prevent the loss of tubule contents during embedding, the tubule sections, were cut well beyond the segment selected for sectioning. For each individual, six sections, each section with 5µm diameter by microtome were cut from tubules. These sections were first placed on gelatin coated slides (the gelatin was heated to 42°c) and then transferred to the oven at 37°c for one hour. This technique usually prevents the fragil tubules from breaking and the loss of gametes. The slides were stained with Eosin and Hematoxylin, and good resolution of the various cell types achieved.A second series of slides was stained with the Periodic Acid Schiff(PAS) to identify polysaccharides(glycogen). Monthly sampling was occurred.The sexual reproductive cycle was defined through the combined use of these criteria: Monthly percentages of the gonad stages for each sex, the monthly gonad index (GI) , given as the ratio of the wet gonad weight (G) to the dray weight (DW)and the monthly percentage of individuals that undetermined sex. The gonad consists of two tufts of tubules on which saccules develop. Gonadal development was classified into five stages: post spawning, recovery, growth, advanced growth, and mature stage that were adapted from the earlier studies of holothurians. Histological preparations showed that the sex of larger individuals could be identified by the presence of oogonia and young oocytes in females, and spermatogonic stages in males.The mean diameter of the tubules and gonadal mass follow annual cycles, increasing from late winter through spring, and dropping abruptly after spawning in the summer. Gametogenesis is generally a prolongate process and begins in March. By summer the ovarian tubules contain oocytes with diameter of 120-240 pm and the testicular tubules contain an abundance of spermatozoa (diameter 5-6 gm ).Following spawning the predominant activity within the spent tubules is phagocytosis of the residual gamets.The active phase of gametogenesis (March to July), coincides with an increasing photoperiod regim, and an accelerated gametogenesis occurs in July when temperature is high. Throughout the year, the gonad of Stichopus hermanni is larger in males than in females, and this is due to the number of tubules in the testis rather than to tubules length or diameter.

Live Attenuated S. Typhimurium Vaccine with Improved Safety in Immuno-Compromised Mice

Live attenuated vaccines are of great value for preventing infectious diseases. They represent a delicate compromise between sufficient colonization-mediated adaptive immunity and minimizing the risk for infection by the vaccine strain itself. Immune defects can predispose to vaccine strain infections. It has remained unclear whether vaccine safety could be improved via mutations attenuating a vaccine in immune-deficient individuals without compromising the vaccine's performance in the normal host. We have addressed this hypothesis using a mouse model for Salmonella diarrhea and a live attenuated Salmonella Typhimurium strain (ssaV). Vaccination with this strain elicited protective immunity in wild type mice, but a fatal systemic infection in immune-deficient cybb-/-nos2-/- animals lacking NADPH oxidase and inducible NO synthase. In cybb-/-nos2-/- mice, we analyzed the attenuation of 35 ssaV strains carrying one additional mutation each. One strain, Z234 (ssaV SL1344_3093), was >1000-fold attenuated in cybb-/-nos2-/- mice and ≈100 fold attenuated in tnfr1-/- animals. However, in wt mice, Z234 was as efficient as ssaV with respect to host colonization and the elicitation of a protective, O-antigen specific mucosal secretory IgA (sIgA) response. These data suggest that it is possible to engineer live attenuated vaccines which are specifically attenuated in immuno-compromised hosts. This might help to improve vaccine safety. © 2012 Periaswamy et al.

Thylakoid terminal oxidases are essential for the cyanobacterium Synechocystis sp. PCC 6803 to survive rapidly changing light intensities.

Cyanobacteria perform photosynthesis and respiration in the thylakoid membrane, suggesting that the two processes are interlinked. However, the role of the respiratory electron transfer chain under natural environmental conditions has not been established. Through targeted gene disruption, mutants of Synechocystis sp. PCC 6803 were generated that lacked combinations of the three terminal oxidases: the thylakoid membrane-localized cytochrome c oxidase (COX) and quinol oxidase (Cyd) and the cytoplasmic membrane-localized alternative respiratory terminal oxidase. All strains demonstrated similar growth under continuous moderate or high light or 12-h moderate-light/dark square-wave cycles. However, under 12-h high-light/dark square-wave cycles, the COX/Cyd mutant displayed impaired growth and was completely photobleached after approximately 2 d. In contrast, use of sinusoidal light/dark cycles to simulate natural diurnal conditions resulted in little photobleaching, although growth was slower. Under high-light/dark square-wave cycles, the COX/Cyd mutant suffered a significant loss of photosynthetic efficiency during dark periods, a greater level of oxidative stress, and reduced glycogen degradation compared with the wild type. The mutant was susceptible to photoinhibition under pulsing but not constant light. These findings confirm a role for thylakoid-localized terminal oxidases in efficient dark respiration, reduction of oxidative stress, and accommodation of sudden light changes, demonstrating the strong selective pressure to maintain linked photosynthetic and respiratory electron chains within the thylakoid membrane. To our knowledge, this study is the first to report a phenotypic difference in growth between terminal oxidase mutants and wild-type cells and highlights the need to examine mutant phenotypes under a range of conditions.

中国淋巴囊肿病毒胸苷酸合酶基因结构特点及分析

胸苷酸合酶(Thymidylate synthase, TS)是进行 DNA 合成所必需的酶类,与细胞分化及肿瘤发生密切相关。淋巴囊肿病毒属于虹彩病毒科成员,是能引起百余种淡、海水鱼感染,并产生肿瘤的病毒病原。在已完成中国淋巴囊肿病毒株 (Lymphocystis disease virus-China, LCDV-C) 基因组序列测定的基础上,本文对位于 LCDV-C 基因组开放阅读框 ORF 011L 的 TS 基因结构、及其推定蛋白结构进行了分析。该基因全长 858bp,GC 含量为 28.2%,编

Effects of tetrabrominated diphenyl ether and hexabromocyclododecanes in single and complex exposure to hepatoma HepG2 cells

This study was designed to determine cytotoxic effects of PBDE-47 and HBCDs individually or with a mixture of both compounds exposure to Hep G2 cells. The results showed PBDE-47 and HBCDs induced increase of nitric oxide synthase (NOS) activity, release of NO. dissipation of mitochondria membrane potential and cell apoptosis. Exposure to HBCDs induced ROS formation. Moreover, preincubation with PTIO (NO scavanger) and N-acetylcysteine (ROS scavanger) partially reversed cytotoxic effects of these compounds. The possible mechanism is that PBDE-47 and HBCDs could boost generation of NO and/or ROS, impact mitochondria, and result in start-ups of apoptosis program. Cells exposed to mixture of both compounds and each of them showed non-apoptotic rate significant difference, but the combination of them caused more adverse effects on cells. These results Suggest that PBDE-47 and HBCDs in single and complex exposure have the cytotoxic activity of anti-proliferation and induction of apoptosis in tumor cells in vitro. (C) 2008 Elsevier B.V. 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.

Effect of dietary carbohydrate-to-lipid ratios on growth and feed utilization in Chinese longsnout catfish (Leiocassis longirostris Gunther)

An 8-week growth trial was carried out in a semi-recirculation system at 26 +/- 0.5 degrees C to investigate the optimal dietary carbohydrate-to-lipid (CHO:L) ratio for carnivorous Chinese longsnout catfish (Leiocassis longirostris Gunther). Triplicate tanks of fish were assigned to each of five isocaloric and isonitrogenous diets with different carbohydrate-to-lipid ratios (0.75, 1.48, 1.98, 2.99 and 5.07). The results showed that a higher specific growth rate (SGR) and feed rate (FR) were observed in the fish fed diet ratios of 1.98 CHO:L (P < 0.05). Overloading dietary carbohydrate (5.07 CHO:L ratio) caused skeletal malformations. Apparent digestibility of dry matter (ADC(d)) significantly increased with dietary CHO:L ratio (P < 0.05), while significantly higher apparent digestibility of protein (ADC(p)) and apparent digestibility of energy (ACD(e)) was observed only in the 1.98 CHO:L group (P < 0.05). Whole body contents of dry matter, lipid and energy significantly increased as the CHO:L ratio decreased (P < 0.05). The hepatosomatic index (HSI) was highest at 1.98 CHO:L ratio (P < 0.05). Highest dietary CHO:L ratio resulted in lower liver glycogen, liver lipid, plasma glucose and plasma triacylglycerol (P < 0.05), whereas there was no significant difference in plasma total cholesterol (P > 0.05). High dietary CHO:L ratio caused pathological changes in fish morphology and liver histology. Based on maximum growth, the optimal carbohydrate-to-lipid ratio was 1.98 for Chinese longsnout catfish.

Effects of hexachlorobenzene on antioxidant status of liver and brain of common carp (Cyprinus carpio)

Hexachlorobenzene (HCB)-induced oxidative damages have been published in rats while the effects have not yet been reported in fishes. Juvenile common carps (Cyprinus carpio) were exposed to waterborne HCB from 2 to 200 mu g l(-1) for 5, 10 or 20 days. Liver and brain were analyzed for various parameters of oxidative stress. There were no significant changes of glutathione (GSH) content and superoxide dismutase (SOD) activity in liver after 5 or 10 days exposure, whereas obvious drops were observed at higher concentrations after 20 days exposure. Significant decreases of GSH content and SOD activity in brain were found during all the exposure days. In brain, HCB also significantly elevated the contents of reactive oxygen species (ROS), thiobarbituric acid-reactive substances (TBARS, as an indicator of lipid peroxidation products), glutathione disulfide (GSSG), and activities of nitric oxide synthase (NOS), glutathione peroxidase (GPx), and glutathione reductase (GR), and inhibited activities of acetylcholinesterase (AchE) and glutathione S-transferase (GST). The results clearly demonstrated that environmentally possible level of HCB could result in oxidative stress in fish and brain was a sensitive target organ of HCB toxicity. (c) 2006 Elsevier Ltd. All rights reserved.

Effects of salt stress on carbohydrate metabolism in desert soil alga Microcoleus vaginatus Gom.

The effects of salt stress on carbohydrate metabolism in Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crusts, were investigated in the present study. Extracellular total carbohydrates and exopolysaccharides (EPS) in the culture medium produced by M. vaginatus increased significantly during the growth phase and reached a maximum during the stationary phase. The production of extracellular carbohydrates also significantly increased under higher salt concentrations, which was attributed to an increase in low molecular weight carbohydrates. In the presence of NaCl, the production of cellular total carbohydrates decreased and photosynthetic activity was impaired, whereas cellular reducing sugars, water-soluble sugars and sucrose content and sucrose phosphate synthase activity increased, reaching a maximum in the presence of 200 mmol/L NaCl. These parameters were restored to original levels when the algae were transferred to a non-saline medium. Sodium and K+ concentrations of stressed cells decreased significantly and H+-ATPase activity increased after the addition of exogenous sucrose or EPS. The results suggest that EPS and sucrose are synthesized to maintain the cellular osmotic equilibrium between the intra- and extracellular environment, thus protecting algal cells from osmotic damage, which was attributed to the selective exclusion of cellular Na+ and K+ by H+-ATPase.

The functional divergence of two glgP homologues in Synechocystis sp PCC 6803

Glycogen phosphorylase (GlgP, EC 2.4.1.1) catalyzes the cleavage of glycogen into glucose-1-phosphate (Glc-1-P), the first step in glycogen catabolism. Two glgP homologues are found in the genome of Synechocystis sp. PCC 6803, a unicellular cyanobacterium: sll1356 and slr1367. We report on the different functions of these glgP homologues. sll1356, rather than slr1367, is essential for growth at high temperatures. On the other hand, when CO2-fixation and the supply of glucose are both limited, slr1367 is the key factor in glycogen metabolism. In cells growing autotrophically, sll1356 plays a more important role in glycogen digestion than slr1367. This functional divergence is also supported by a phylogenetic analysis of glgP homologues in cyanobacteria.

Effect of dietary carbohydrate sources on growth performance and utilization for gibel carp (Carassius auratus gibelio) and Chinese longsnout catfish (Leiocassis longirostris Gunther)

The nutritional function of monosaccharides, disaccharides and polysaccharides for omnivorous gibel carp and carnivorous Chinese longsnout catfish were investigated and the ability of these two species to utilize carbohydrates was compared. For each species, triplicate groups of fish were assigned to each of five groups of isoenergetic and isonitrogenous experimental diets with different carbohydrate sources: glucose, sucrose, dextrin, soluble starch (acid-modified starch) and alpha-cellulose. The carbohydrates were included at 60 g kg(-1) in Chinese longsnout catfish diets and at 200 g kg(-1) in gibel carp diets. A growth trial was carried out in a recirculation system at 27.8 +/- 1.9 degrees C for 8 weeks. The results showed that fish with different food habits showed difference in the utilization of carbohydrate sources. For gibel carp, better specific growth rate (SGR) and feed efficiency (FE) were observed in fish fed diets containing soluble starch and cellulose, but for Chinese longsnout catfish, better SGR and FE were observed in fish fed diets containing dextrin and sucrose. Apparent digestibility coefficient of dry matter (ADC(d)) and apparent digestibility coefficient of energy (ADC(e)) were significantly affected by dietary carbohydrate sources in gibel carp. ADC(d) and ADC(e) significantly decreased as dietary carbohydrate complexity increased in Chinese longsnout catfish except that glucose diet had medium ADC(d) and ADC(e). In both species, no significant difference of apparent digestibility coefficient of protein was observed between different carbohydrate sources. Dietary carbohydrate sources significantly affected body composition, and liver phosphoenolpyruvate carboxykinase (PEPCK), pyruvate kinase (PK), glucose 6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) activities also varied according to dietary carbohydrate complexity. Fish with different food habits showed different abilities to synthesize liver glycogen, and the liver glycogen content in gibel carp was significantly higher than in Chinese longsnout catfish. The influence of carbohydrate source on gluconeogenesis and lipogenesis was also different in the two fish species.

Salt tolerance of Microcoleus vaginatus Gom., a cyanobacterium isolated from desert algal crust, was enhanced by exogenous carbohydrates

Microcoleus vaginatus isolated from a desert algal crust of Shapotou was cultured in BG-11 medium containing 0.2mol l(-1) NaCl or 0.2mol l(-1) NaCl plus 100mg l(-1) sucrose, extracellular polymeric substances (EPS) or hot water-soluble polysaccharides (HWP), respectively. Photosynthetic oxygen evolution rates, photosystem 11 activity (Fv/Fm) and dark respiration of NaCl-stressed cells were enhanced significantly by the added sucrose or EPS under salt stress conditions (0.2mol l(-1) NaCl). Compared with cells treated with salt alone, sodium contents in cells reduced significantly; the content of cellular total carbohydrate did not change, and intracellular sucrose, water-soluble sugar increased significantly following the addition of exogenous carbohydrates. Sucrose synthase (SS) activity of NaCl-stressed cells increased following the addition of sucrose, and sucrose phosphate synthase (SPS) activity of NaCl-stressed cells increased following the addition of exogenous sucrose, EPS or HWP compared with cells stressed with NaCl only. The results suggested that the extruded EPS might be re-absorbed by cells of M. vaginatus as carbon source, they could increase salt tolerance of M. vaginatus through the changes of carbohydrate metabolism and the selective uptake of sodium ions. (C) 2003 Elsevier Science Ltd. All rights reserved.

Selection and ultrastructural observation of a high-CO2-requiring mutant of cyanobacterium Synechococcus sp PCC7942

A high-CO2-requiring mutant of Synechococcus sp. PCC7942 las been isolated after chemical mutagenesis of ethyl methane sulphonate (EMS). It was able to grow at 4% CO2, but not under ambient CO2. The initial screening of the mutant showed that the genetic reversion rate was about 10(-7) and death occurred 2 -3 days after being transferred from 4% CO2 to the ambient air. Its photosynthetic dependence on external dissolved inorganic carbon was higher than that of the wild type cells, but its carbonic anhydrase activity was comparatively low. In the ultrastructural level, various types of aberrant carboxysomes appeared in the mutant cells: rod-shaped carboxysomes, irregular carboxysomes and the "empty-inclusion carboxysomes" with increasing number of glycogen granules surrounding the thylakoids. All these alterations indicated that the mutant was defective in utilizing the external CO2. The induction of carboxysomes by lower levels of CO2 and the biogenesis of carboxysomes are herein discussed.

RNAi沉默烟草GA 20-氧化酶基因研究

赤霉素是一种高效能的广谱植物生长调节剂，为五大植物激素之一，具有重要的生物学功能。目前利用赤霉素突变体研究生物合成途径和信号转导已经成为热点。 GA 20-氧化酶是GA生物合成中的一类关键酶，它位于GA合成途径的中心位置。本研究根据烟草（Nicotiana tabacum）GA 20-氧化酶基因序列，设计2对分别含有特定酶切位点的特异引物，以烟草基因组DNA为模板，扩增目的基因(约250 bp)片段。将正、反向目的片段分别插入中间载体的内含子两侧，再经BamH I和Sac I双酶切回收约700 bp的目的片段，插入到双元载体质粒p2355中，成功构建了含GA 20-氧化酶基因片段反向重复序列的植物表达载体p23700。分别将p2355质粒和p23700质粒导入根癌农杆菌（Agrobacterium tumefaciens）EHA105中并转化烟草叶片细胞，经卡那霉素选择培养，PCR及GUS组织染色鉴定，获得转基因烟草植株。以EHA105-p2355转化的烟草，获得41株转基因植株，均没有矮化表型；而以EHA105-p23700转化的烟草，获得转基因植株14株，其中具有矮化表型的烟草10株，表明反向重复序列转录产物能形成发夹RNA(hpRNA)，产生小分子干扰RNA（small interferring RNA，简称siRNA），干扰目的基因的表达。 赤霉素含量测定表明矮化植株中赤霉素合成途径的最终产物GA3总含量明显低于野生型烟草植株。荧光定量PCR结果表明，矮化转基因烟草的GA 20-氧化酶基因表达量受到明显抑制，表达量明显低于野生型对照。同时对上游内根-贝壳杉合成酶（Ent-kaurene synthase，KS）基因，下游的GA-3β羟化酶基因进行了RT-PCR分析，结果显示上游基因的表达没有规律性变化，而下游基因表达量亦降低。上述结果表明，GA 20-氧化酶基因的表达被有效地干扰了，表达受到抑制，从而影响植株体内GA3的合成，影响植株的生长发育，导致植株矮化。并推测，GA 20-氧化酶基因受到抑制，可能影响下游基因的表达。并且通过干旱胁迫测试，发现矮化植株相对于野生型植株及不含干扰片段的转基因植株，对干旱的耐受力有了很大的提高，具有更强的耐受力。 研究结果为进一步进行相关研究奠定基础。 Gibberellin(GA) is an efficient plant growth regulator. As one of five major plant hormones, it plays an important biological function. Using GA mutant for investigating biosynthetic pathways and signal transduction has become high lights. GA 20-oxidase is a crucial enzyme involved in gibberellin biosynthesis. According to tobacco (Nicotiana tabacum) GA 20-oxidase enzyme gene sequence and based on binary vector p2355, we constructed a plant expression vector p23700, which habors an inverted repeat DNA fragment of GA 20-oxidase gene drivered by Cauliflower mosaic virus promtor (CaMV 35Sp). Binary plasmid p2355 had no inverted repeat DNA fragment of GA 20-oxidase gene. The vector p2355 and p23700 were introduced into Agrobacterium tumefaciens EHA105 and tobacco leaf transformation was conducted. After selected by kanamycin and characterized by PCR and GUS hischemical reaction, transsgenic plants were obtained. Fourtheen transgenic plants, which were transformed by EHA105-p23700, were obtained. Among them, 10 were dwarf mutants. However, 41 transgenic plants with the same normal phenotype as wild type,which were transformed by EHA105-p2355, were obtained. Analysis of Gibberellin contents showed that it was lower in dwarf mutants than in normal phenotype plants. Moreover, comparing to normal phenotype plants including wild type and transgenic plants with no interference fragment, the drought tolerance of dwarf plants have greatly increased. And their proline content increased obviously after drought test. Fluorescence quantitative real time PCR (RT-PCR) showed that GA 20-oxidase gene expression was significantly inhibited in dwarf transgenic tobacco. Meanwhile, the expression of the upstream gene ent-kaurene synthase (KS) gene and downstream gene GA-3β hydroxylase gene was also detected by RT-PCR. The results presented that KS gene expression had no regular change while GA-3β hydroxylase gene expression reduced. It implied that inhibiting GA 20-oxidase gene probably reduce the expression of downstream genes. The results showed that the transcriptional products of the foreign inverted repeat fragment can form hairpin RNA (hpRNA) to induce RNAi. It presented that GA 20-oxidase gene expression was effectively interfered, resulting in reducing GA3 synthesis and inhibiting plant growth and development, then dwarf plants were produced. However, the dwarf plants had higher tolerance of drought.

辐射诱导的HeLa细胞旁效应信号通路研究

随着肿瘤认识的不断深入，肿瘤外科学、肿瘤放射治疗学、肿瘤化学治疗学构成了现代肿瘤治疗学的三大支柱，而放射治疗学的研究对于肿瘤的治疗有重要的临床意义。本文通过对HeLa进行辐射来观察其产生的旁效应信号通路，从而对临床工作的起到一定的帮助。辐照过的细胞通过释放信号分子引起周围未辐照细胞产生一系列的生物学反应的现象，称之为辐射诱导的旁效应。活性氧（Reactive oxygen species, ROS）和一氧化氮（Nitric oxide, NO）是辐射诱导的旁效应信号通路中的两个重要信号分子。本文研究了这两种重要的信号分子在辐射诱导的HeLa细胞旁效应信号通路中的关系。通过微核实验，我们发现X射线以及12C6+ 辐照过的HeLa细胞及其旁观者细胞的微核形成明显增加，而1%的二甲亚砜（Dimethyl sulfoxide，DMSO，ROS清除剂）预处理X射线辐照的细胞则显著抑制了受辐照细胞及其旁观者细胞的微核形成。1 Gy的X射线辐照能够抑制细胞的增殖速率而0.5%和1%的DMSO预处理则能减少X射线的增殖抑制作用，并且DMSO预处理的效果与浓度有关：1%的DMSO比0.5%的DMSO处理更大程度的恢复了受辐照细胞的增殖速率。另一方面，接受条件培养基（Conditioned medium）的旁观者细胞的增殖速率增加，而DMSO预处理产生条件培养基的受辐照细胞则使旁观者细胞的增殖速率降低，且DMSO预处理的效果同样与其浓度相关：浓度越高，条件培养基的刺激生长作用越小。Western blotting和DAF-FM DA荧光探针检测分别显示了辐照过后细胞的诱导型一氧化氮合酶（Inducible nitric oxide synthase, iNOS）和NO水平均升高，而DMSO预处理则降低其水平。因此，我们推测在X射线辐照的HeLa细胞旁效应信号通路当中ROS是NO的上游信号。另外，我们采用培养基转移后立即加DMSO或BMS－345541（IKK/NF-κB抑制剂）的方法研究了旁观者细胞当中的旁效应信号通路。我们发现DMSO和BMS－345541均显著抑制了旁观者细胞的NO水平。因此，在旁观者细胞当中ROS与NF-κB均为NO的上游信号