诱导型Hsp70在外源性甲醛对小鼠肝肾氧化损伤中的保护作用

为探讨外源性甲醛是否会对小鼠肝脏和肾脏的热休克蛋白70(heat shock protein 70, Hsp70)产生影响,选雄性Balb/c小鼠为研究对象,采用动态吸入方式染毒2周,每周5 d,每天8 h,随机分为空白对照组、3 mg·m-3甲醛染毒组。染毒结束后,计算肝脏和肾脏的脏体比,检测各器官中活性氧(reactive oxygen species, ROS)、丙二醛(malondialdehyde, MDA)、还原型谷胱甘肽(glutathione, GSH)的含量,同时用实时荧光定量PCR(real-time PCR, RT-PCR)法检测Hsp70的基因表达,并通过HE染色观察形态学变化。结果表明:与对照组相比,2种器官的脏体比均显著下降(P0.05或P0.01); ROS、MDA含量上升、GSH含量下降,出现显著性差异(P0.05或P0.01); Hsp70基因表达水平下降且差异显著(P0.05或P0.01); HE切片显示,甲醛暴露使肝脏的中央静脉窦的破损程度增加、细胞核变大,肾脏中肾小中肾小球基底膜变窄、体积缩小甚至分解。上述结果表明,甲醛暴露会对小鼠的的肝脏和肾脏造成不同程度的氧化损伤,小鼠可通过上调Hsp70的基因表达以增强机体的抗氧化能力。     

根癌农杆菌介导的高赖氨酸蛋白基因转化叶用莴苣的研究

构建了植物表达载体pLBI，该载体携带35S启动子、高赖氨酸蛋白基因(LRP)、NPTⅡ基因和NOS终止子，对影响根癌农杆菌(Agrobacterium tumefaciens L.)转化的多种因素进行探索后，建立了一种农杆菌介导的稳定的叶用莴苣遗传转化系统．选择目前生产上大量推广应用的叶用莴苣品种，以其无菌苗子叶为外植体，经农杆菌LBA4404(含质粒pLBI)感染，在含ρ(Kan)／mgL^-1=100的芽分化培养基上筛选转化芽，转入含相同浓度抗生素的生根培养基上进行生根筛选，直至得到完整的再生植株．PCR扩增及Southern blot的检测结果均表明，高赖氨酸蛋白基因已整合到叶用莴苣基因组中．图4表4参6     

Role of NADH-dependent chromium reductases,exopolysaccharides and antioxidants by Paenibacillus thiaminolyticus PS 5 against damage induced by reactive oxygen species

ABSTRACT

Cr (VI) being used in various activities of industries and its improper treatment lead to contamination of environment. Among different methods, biological is the most efficient method to control pollution from soils affected with metals. Present study was designed to assess the role of Paenibacillus thiaminolyticus PS5 for adsorption, Cr (VI) reduction and mechanism of reduction. Sorption of chromium (VI) by strain PS5 was obtained by batch equilibrium method. Cr (VI) reduction in both free and immobilised cells were evaluated by 1,5-Diphenyl Carbazide method. The formation of biofilm by Paenibacillus thiaminolyticus PS5 was observed for colour change as well as quantified spectrophotometrically. Analysis kits were used to measure the amount of eDNA, superoxide and malondialdehyde (MDA). Metal resistant strain PS5 was characterised as P. thiaminolyticus using 16S rRNA gene sequence. Maximum biosorption of Cr (VI) by strain PS5 was found at pH 6–8 and 100–400 µgCr/mL within 24 hours of incubation. Complete reduction of Cr (VI) by strain PS5 was achieved at pH 6–8 and100–300 µg/mL Cr (VI). Paenibacillus thiaminolyticus PS5 immmobilisation on sodium alginate and polyvinyl alcohol facilitated complete reduction of Cr (VI) within 18 hours due to the formation of more biofilm under metal stress conditions. Strain PS5 reduced almost all Cr (VI) into Cr (III) in supernatant, most of which was immobilised by cell debris. Experiments confirmed the reduction of Cr (VI) by cytosolic cell-free extracts which is a mechanism of detoxification. The release of exopolysaccharides and antioxidants by strain PS5 played a protective role against cell damage by Cr (VI) as Cr (VI) could not release the significant amount of eDNA, superoxide and MDA. The present study proved strain PS5 to be a super bioinoculant which has great capacity for adsorption, biotransformation and can activate cytosolic reductases, exopolysaccharides and antioxidants against oxidative damage.     

城市生活污水处理中剩余污泥处理技术探讨

分析了武汉市污水处理现状、剩余污泥的产量与处理技术概况,介绍了制备蛋白质泡沫灭火剂技术再利用剩余污泥的可行性。     

磷酸化蛋白质组学技术的发展及其在环境毒理研究中的应用

磷酸化修饰是蛋白质最主要的翻译后修饰形式之一,磷酸化蛋白质组学从整体上观察细胞或组织中磷酸化修饰的状态及其变化,为探讨药物刺激和环境应激下生物体受损的生物学过程提供新的视角.针对磷酸化蛋白质组学技术的发展以及其在环境毒理研究中的应用展开综述.首先,从磷酸化肽富集、磷酸化蛋白的鉴定和磷酸化位点的预测、定量磷酸化蛋白质组学研究3个方面对磷酸化蛋白质组学技术的研究内容和分析策略进行了概述.在此基础上,按照离体实验、活体实验以及毒性作用通路分析3部分对磷酸化蛋白质组学在环境毒理研究中的应用进行了详细阐述.最后,总结了目前磷酸化蛋白质组学研究的不足,并有针对性地提出了磷酸化蛋白质组学在环境毒理研究中的发展方向.     

Bioaugmentation of DDT-contaminated soil by dissemination of the catabolic plasmid pDOD

A plasmid transfer-mediated bioaugmentation method for the enhancement of dichlorodiphenyltrichloroethane (DDT) degradation in soil was developed using the catabolic plasmid pDOD from Sphingobacterium sp. D-6. The pDOD plasmid could be transferred to soil bacteria, such as members of Cellulomonas, to form DDT degraders and thus accelerate DDT degradation. The transfer efficiency of pDOD was affected by the donor, temperature, moisture, and soil type. Approximately 50.7% of the DDT in the contaminated field was removed 210 days after the application of Escherichia coli TG I (pDOD-gfp). The results suggested that seeding pDOD into soil is an effective bioaugmentation method for enhancing the degradation of DDT.     

Structural and molecular alterations in arsenic-induced hepatic oxidative stress in rats: a FTIR study

Arsenic (As) is an ubiquitously distributed environmental toxicant predominantly contaminating drinking water. A number of studies indicated that oral exposure of humans to inorganic As produced damage to various body tissues including liver. Oxidative stress is thought to play a major role in As-induced hepatotoxicity. In this study, Fourier transforms infrared (FTIR) spectroscopy approach was applied to determine whether chronic As exposure at 25 ppm, intragastrically for 12 weeks, affected oxidative stress status in rat liver. Data demonstrated that chronic As administration exacerbated oxidative stress as was evidenced by suppressed antioxidant defense system and increased lipid peroxidation and protein oxidation. The FTIR study showed that peak area value of amide A decreased significantly followed by reduced amide I and amide II peak area in an As-treated rat liver suggesting altered protein profile. The change of Olefinic?CH stretching band and C?O stretching of triglycerides band indicated the altered lipid levels due to metallic exposure. The fall in the peak area of PO₂^? asymmetric stretching in the As-treated group might be due to compositional changes of nucleic acids. Hence, the results of this study indicate that As-induced oxidative stress was associated with structural and molecular modifications in proteins, lipids, and glycogen in a rat liver that may help to elucidate molecular mechanisms underlying metal-mediated hepatic damage.     

Endogenous nitric oxide mediates alleviation of cadmium toxicity induced by calcium in rice seedlings

The effect of calcium chloride (CaCl₂) on rice seedling growth under cadmium chloride (CdCl₂) stress, as well as the possible role of endogenous nitric oxide (NO) in this process, was studied. The growth of rice seedlings was seriously inhibited by CdCl₂, and the inhibition was significantly mitigated by CaCl₂. However, hemoglobin (Hb) and 2-(4-carboxyphenyl)-4, 4,5,5-tetramethylimidazolinel- oxyl-3-oxide (cPTIO) weakened the promotion effect of CaCl₂. The results of NO fluorescence localization suggest that growth accelerated by CaCl₂ might be associated with elevated NO levels. The content of Cd, protein thiols (PBT), and nonprotein thiols (NPT) in cell walls, cell organelles, and soluble fractions, respectively, of rice seedlings decreased considerably in the presence of CaCl₂, whereas the content of pectin, hemicellulose 1 (HC1), and hemicellulose 2 (HC2) increased significantly. Elimination of endogenous NO in Cd+Ca treatment could promote the transportation of Cd²⁺ to cell organelles and soluble fractions and increase the content of NPT and PBT in leaves. In addition, transportation of Cd²⁺ to cell organelles and soluble fractions was retarded in roots, the content of NPT increased, and the content of PBT decreased. With elimination of endogenous NO in Cd+Ca treatment, the content of pectin, HC1, and HC2 decreased significantly. Thus, Ca may alleviate Cd toxicity via endogenous NO with variation in the levels of NPT, PBT, and matrix polysaccharides.