Oxidative stress-resistance assay for screening yeast strains overproducing heterologous proteins

Many natural proteins have been developed into drugs and produced for direct application. Identifying improved hosts to achieve high-level heterologous protein production is a challenge in the study of heterologous protein expression in recombinant yeast. In this study, a novel high-throughput assay to screen such overproducing Saccharomyces cerevisiae strains was systematically developed. The protocol designed was based on screening host strain derivatives with increased superoxide dismutase dependent resistance to oxidative stress. Yeast cells transformed with recombinant plasmid carrying SOD1 gene as a reporter responded exquisitely to oxidative stress induced by elevated concentrations of paraquat. Improved yeast strains resulting from screening clones subjected to genome shuffling through selective pressure argue for a more effective screening system compared with traditonal selection. Moreover, this approach can be employed in general biochemical analysis without utilization of flow cytometry or well plate reader. Therefore, it is expected that the high-throughput assay would make superior strains producing heterologous proteins.     

Waterbird habitat-selection during winter and spring in reclaimed coastal wetlands in Nanhui Dongtan, Shanghai

From December 2009 to May 2010 goose and duck (Anatidae) community censuses in winter and shorebird (Charadriiforms) community censuses in spring were conducted across three types artificial wetlands (urban lake wetland, restorative wetland, abandoned wetland) along the coast of Nanhui, Shanghai. Correlation analyses were undertaken between community indices and habitat factors. The results showed there were significant differences in the density of geese and ducks among the wetlands, but no difference in the number of species. The density of geese and ducks in the restorative wetland was 3.77 times that of abandoned wetland and 6.03 times that of urban lake wetlands. The number of species and density of shorebirds in restorative wetlands was 2.88 and 5.70 times that of abandoned wetlands. We found significant differences in the number and density of shorebird species between restorative and abandoned wetlands. The number of species density of geese and ducks and the Shannon-Wiener (H') index were positively correlated with water area. The number of species and H' were negatively correlated with vegetation area. The number of species, species density and H' and evenness were negatively correlated with vegetation coverage. H' was positively correlated with mean water level. The results showed that the number and density of shorebird species were positively correlated with bare muddy areas. Aquaculture ponds and paddy fields in reclaimed area is efficient sufficient compensation mechanism to maintain more water areas for waterbirds and to control vegetation expansion and maintain shorebird habitat after coastal reclamation.     

阿米洛利抑制NHE-1减轻低氧性肺动脉平滑肌细胞增殖

目的：研究Na^＋／H^＋交换抑制剂阿米洛利对低氧刺激的大鼠肺动脉平滑肌细胞（PASMCs）增殖的影响,以及Na^＋／H^＋交挟体-l（NHE-1）活性和表达的变化.方法：常氧（21％O2）或低氧（2％O2）条件下培养PASMCs,并分别给予浓度为1．653、3．125、6．25、12．5、25和50μmol／L．等不同浓度的阿米洛利,培养24h,采用MTT比色实验和免疫组化检测PCNA阳性细胞率的方法反映细胞增殖情况,同时采用激光共聚焦检测细胞内pH以反映Na^＋／H^＋交换体-1活性,RT—PCR法检测Na^＋／H^＋交换体-1mRNA的表达量.结果：低氧培养的PASMCs细胞内pH升高,NHE—1mRNA的表达增多,而阿米洛利可以降低细胞内pH,减少NHE—1mRNA的表达量。同时低氧较常氧培养MTT光吸收值较常氧培养明显升高。PCNA阳性细胞率明显增高,而给予阿米洛利时上述两个指标随药物浓度增加而逐渐下降。结论：低氧可以激活PASMCs细胞膜上的E—1,增加其mRNA水平表达量,使细胞内碱化,促进细胞增殖,而Na^＋／H^＋交换抑制剂阿米洛利可以抑制其活性,减少mRNA水平的表达,导致细胞内酸化,从而抑制细胞增殖,并且此抑制作用在3．125～50μmol／L．浓度范围内呈现明显的浓度依赖性。     

Protective effects of pre-germinated brown rice diet on low levels of Pb-induced learning and memory deficits in developing rat

Lead (Pb) is a known neurotoxicant in humans and experimental animals. Numerous studies have provided evidence that humans, especially young children, and animals chronically intoxicated with low levels of Pb show learning and memory impairments. Unfortunately, Pb-poisoning cases continue to occur in many countries. Because the current treatment options are very limited, there is a need for alternative methods to attenuate Pb toxicity. In this study, the weaning (postnatal day 21, PND21) rats were randomly divided into five groups: the control group (AIN-93G diet, de-ionized water), the lead acetate (PbAC) group (AIN-93G diet, 2 g/L PbAC in de-ionized water), the lead acetate + WR group (white rice diet, 2 g/L PbAC in de-ionized water; PbAC + WR), the lead acetate + BR group (brown rice diet, 2 g/L PbAC in de-ionized water; PbAC + BR) and the lead acetate + PR group (pre-germinated brown rice diet, 2 g/L PbAC in de-ionized water; PbAC + PR). The animals received the different diets until PND60, and then the experiments were terminated. The protective effects of pre-germinated brown rice (PR) on Pb-induced learning and memory impairment in weaning rats were assessed by the Morris water maze and one-trial-learning passive avoidance test. The anti-oxidative effects of feeding a PR diet to Pb-exposed rats were evaluated. The levels of reactive oxygen species (ROS) were determined by flow cytometry. The levels of 8-hydroxy-2-deoxyguanosine (8-OHdG), γ-aminobutyric acid (GABA) and glutamate were determined by HPLC. Our data showed that feeding a PR diet decreased the accumulation of lead and decreased Pb-induced learning and memory deficits in developing rats. The mechanisms might be related to the anti-oxidative effects and large amount of GABA in PR. Our study provides a regimen to reduce Pb-induced toxicity, especially future learning and memory deficits in the developing brain.     

牡丹江鱼类单殖吸虫一新记录

近年来,三代虫在黑龙江省各虹鳟鱼场普遍发生,为害程度与日俱增。       

外来植物入侵的全境性研究进展与展望

由于外来种入侵地一般都远离其自然分布区(原产地), 如果只在其入侵地或者原产地进行研究, 很难真正发现其入侵性形成和成功入侵的根本原因。目前, 许多学者开始关注和倡导对入侵种在原产地和入侵地的表现同时进行研究, 即入侵种的全境性研究(whole-range studies), 为入侵生物现有地理分布格局的形成原因和入侵机制等提供解释。本文结合国内外关于入侵植物全境性研究的进展和成果, 分别针对研究的主要目的、内容、意义等进行了全面的阐述, 探讨了存在的问题与不足, 并对未来相关研究进行了展望。目前已有的全境性研究主要是通过野外直接观测和同质种植园实验来比较入侵种在入侵地和原产地的生长、繁殖和生理生态等表型性状的差异, 以及应用分子标记方法比较入侵地种群和原产地种群遗传多样性的差异, 进行入侵植物的分子系统地理学研究, 从而有效检验生物入侵机制的理论和假说, 深入阐明植物入侵的机制, 为制定入侵植物的防控策略提供指导。值得注意的是, 由于外来植物入侵的全境性研究起步较晚, 现有研究的方法和内容还不够完善, 今后需要在加强国际合作的基础上进一步改善。     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆(Glycine max)初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200 μmol·L^–1 的CuSO₄溶液进入根中, 然后在重力作用下从根基部灌注400 μmol·L^–1的K₄[Fe(CN)₆]溶液, 两种物质在根内相遇即可产生棕色的Cu₂[Fe(CN)₆]沉淀, 根据沉淀的位置来确定铜离子所经过的途径。结果表明: Cu²⁺可以穿过内皮层凯氏带, 在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀, 侧根发生的部位也产生了大量的沉淀; 当抽取K₄[Fe(CN)₆]溶液后再灌注CuSO₄溶液, 发现Cu²⁺仍然可以穿过凯氏带, 并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

Transcriptome Profiles in the Spleen of the Chinese Giant Salamander (Andrias davidianus) Challenged with Citrobacter freundii

Russian Journal of Bioorganic Chemistry - The Chinese giant salamander (Andrias davidianus) is one of the most important ecological breeding species with distinct characteristics and is cultured in...     

Noninvasive ultrasound deep brain stimulation of nucleus accumbens induces behavioral avoidance

Ultrasound stimulation is an emerging noninvasive option in treating neuropsychiatric disorders. The present study investigates the behavioral alterations resulting from ultrasound stimulation on the nucleus accumbens(NAc) in freely moving mice. Our results show that an acute ultrasound stimulation on the NAc, rather than the visual cortex or auditory cortex, led to a pronounced avoidance behavior, while repeated NAc ultrasound stimulation resulted in an obvious conditioned place aversion with changes in synaptic protein(Glu A1/2 subunit) expression. Notably, NAc ultrasound stimulation suppressed the morphine-induced conditioned place preference. The results provide evidence that NAc ultrasound stimulation can be applied as a potential noninvasive therapeutic option in treating psychiatric disorders.