一株受二噁英类化合物诱导表达绿色荧光蛋白的胃癌细胞系的建立

将人工合成的5个相连的XRE(Xeobiotic response element)和minimal CMV(Human cytomegalovirus)启动子,插入载体pEGFP-1,构建得到载体pXRE5-EGFP.以pXRE5-EGFP转染人胃癌细胞SGC-7901,经过筛选得到一株受2,3,7,8-四氯代二苯并二英(2,3,7,8-tetrachlorodibenzo-p-dioxin,TCDD)诱导表达绿色荧光蛋白的细胞系XRE5-SGC-7901.将XRE5-SGC-7901细胞株暴露于不同浓度TCDD(0、250、500pg·mL-1)中,发现TCDD暴露组XRE5-SGC-7901细胞中绿色荧光蛋白的表达量显著升高,且与TCDD暴露浓度呈正相关.新构建的细胞株具有二英类化合物诱导表达绿色荧光蛋白的功能.     

Predicting the mating system from phenotypic correlations between life-history and sperm quality traits in the Alpine whitefish <Emphasis Type="Italic">Coregonus zugensis</Emphasis>

The mating behavior and reproductive strategies of Alpine whitefish like Coregonus zugensis (Nüsslin) are poorly understood, probably because they spawn in deep water where direct observations are difficult. In this study, we interpret life-history and sperm quality traits of fish that we caught from their spawning place. We found that males invest heavily into gonadal tissue (up to 5.6% of their body weight), which is, in comparison to other fish, consistent with external fertilization, distinct pairing and moderate to high communal spawning, or no pairing and low to moderate communal spawning. Sperm competition theory and recent experimental studies on other salmonids predict that males optimize ejaculate characteristics in relation to the costs of sperm and the level of competition they have to expect: dominant males are predicted to invest less into ejaculate quality and to have slower spermatozoa than subdominant males. We found that spermatozoa of older males are slower than those of younger males. Moreover, older males have larger breeding tubercles, a secondary sexual trait that has, in some previous studies, been found to be linked to good condition and to good genetic quality. Our results suggest that C. zugensis has age-linked reproductive strategies, that multimale spawning is common, i.e., that sperm competition plays a significant role, and that older males are on average dominant over younger males at the spawning place.     

毒理基因组学研究进展

毒理基因组学是利用最新的基因组学技术来进行毒理学研究的新兴学科。它能够快速全面地检测出化合物和生物体相互作用后的全基因组表达的变化,再通过生物信息学的方法对化合物的毒性进行定性分析。它可以为传统毒理学检测筛选更多的生物学标志物,解释有毒物质的致毒机理,降低风险评价的不确定性。但是,毒理基因组学还存在许多问题：如实验设计不统一,分析理论不完善,检测费用太昂贵等。其中最主要的理论问题在于,虽然人们早已认识到毒理基因组学最大的优势是可对毒性进行较全面的分类和预测,但是已有的研究仍然集中在生物标志物的筛选和致毒机理的解释上,而没有充分利用全基因组变化的信息,因为理论上除了实验误差引入的基因表达变化以外的变化都是化合物和生物体相互作用的结果。要解决这些问题,新发现的microRNA毒理基因组和动态基因表达图谱的研究可能是一个潜在方向。总体来说,目前毒理基因组学只能作为风险评价的参考,但是它最终将为风险评价提供有力的理论依据和准确预测,提高风险评价的可靠性。     

Influences and mechanisms of nanofullerene on the horizontal transfer of plasmid-encoded antibiotic resistance genes between E. coli strains

• Sub-inhibitory levels of nC₆₀ promote conjugative transfer of ARGs. • nC₆₀ can induce ROS generation, oxidative stress and SOS response. • nC₆₀ can increase cell membrane permeability and alter gene expression. • Results provide evidence of nC₆₀ promoting antibiotic resistance dissemination. The spread and development of antibiotic resistance globally have led to severe public health problems. It has been shown that some non-antibiotic substances can also promote the diffusion and spread of antibiotic resistance genes (ARGs). Nanofullerene (nC₆₀) is a type of nanomaterial widely used around the world, and some studies have discovered both the biological toxicity and environmental toxicity of nC₆₀. In this study, cellular and molecular biology techniques were employed to investigate the influences of nC₆₀ at sub-minimum inhibitory concentrations (sub-MICs) on the conjugation of ARGs between the E. coli strains. Compared with the control group, nC₆₀ significantly increased the conjugation rates of ARGs by 1.32‒10.82 folds within the concentration range of 7.03‒1800 mg/L. This study further explored the mechanism of this phenomenon, finding that sub-MICs of nC₆₀ could induce the production of reactive oxygen species (ROS), trigger SOS-response and oxidative stress, affect the expression of outer membrane proteins (OMPs) genes, increase membrane permeability, and thus promote the occurrence of conjugation. This research enriches our understanding of the environmental toxicity of nC₆₀, raises our risk awareness toward nC₆₀, and may promote the more rational employment of nC₆₀ materials.     

Optimized determination of airborne tetracycline resistance genes in laboratory atmosphere

• Sampling parameters with high efficiency was determined. • Operational process to detect airborne ARGs was optimized. • Providing research basis to control airborne ARGs of a laboratory atmosphere Antibiotic resistance genes (ARGs) have been detected in various atmospheric environments. Airborne ARGs transmission presents the public health threat. However, it is very difficult to quantify airborne ARGs because of the limited availability of collectable airborne particulate matter and the low biological content of samples. In this study, an optimized protocol for collecting and detecting airborne ARGs was presented. Experimental results showed that recovery efficiency tended to increase initially and then declined over time, and a range of 550–780 copies/mm² of capture loading was recommended to ensure that the recovery efficiency is greater than 75%. As the cell walls were mechanically disrupted and nucleic acids were released, the buffer wash protects ARGs dissolution. Three ratios of buffer volume to membrane area in buffer wash were compared. The highest concentrations of airborne ARGs were detected with 1.4 µL/mm² buffer wash. Furthermore, the majority of the cells were disrupted by an ultrasonication pretreatment (5 min), allowing the efficiency ARGs detection of airborne samples. While, extending the ultrasonication can disrupt cell structures and gene sequence was broken down into fragments. Therefore, this study could provide a theoretical basis for the efficient filter collection of airborne ARGs in different environments. An optimized sampling method was proposed that the buffer wash was 1.4 µL/mm² and the ultrasonication duration was 5 min. The indoor airborne ARGs were examined in accordance with the improved protocol in two laboratories. The result demonstrated that airborne ARGs in an indoor laboratory atmosphere could pose the considerable health risk to inhabitants and we should pay attention to some complicated indoor air environment.     

传递系数法在滑坡稳定性评价中的几个问题探讨

本文运用LASA软件,分析了采用传递系数法计算滑坡稳定系数时,传递系数计算式、条块划分、渗透压力计算对稳定系数的影响。传递系数计算式的分析得出采用迭代法求解是比较合理的方法;通过实例对条块划分导致的影响分析,可知条块划分应当尽量与滑体的实际情况相符合才可以减小计算误差;渗透压力计算分析指出稳定性计算考虑动水压力时,应当根据不同情况选择不同水力坡降计算式。     

Metagenomic insights into responses of microbial population and key functional genes to fulvic acid during partial nitritation

The long-term impact of fulvic acid (FA) on partial nitritation (PN) system was initially examined in this study. The obtained results revealed that the FA lower than 50 mg/L had negligible effect on the nitrite accumulation rate (NAR nearly 100%) and ammonium removal rate (ARR 56.85%), while FA over 50 mg/L decreased ARR from 56.85% to 0.7%. Sludge characteristics analysis found that appropriate FA (<50 mg/L) exposure promoted the settling performance and granulation of PN sludge by removing Bacteroidetes and accumulating Chloroflexi. The analysis of metagenomics suggested that the presence of limited FA (0-50 mg/L) stimulated the generation of NADH, which favors the denitrification and nitrite reduction. The negative impact of FA on the PN system could be divided into two stages. Initially, limited FA (50-120 mg/L) was decomposed by Anaerolineae to stimulate the growth and propagation of heterotrophic bacteria (Thauera). Increasing heterotrophs competed with AOB (Nitrosomonas) for dissolved oxygen, causing AOB to be eliminated and ARR to declined. Subsequently, when FA dosage was over 120 mg/L, Anaerolineae were inhibited and heterotrophic bacteria reduced, resulting in the abundance of AOB recovered. Nevertheless, the ammonium transformation pathway was suppressed because genes amoABC and hao were obviously reduced, leading to the deterioration of reactor performance. Overall, these results provide theoretical guidance for the practical application of PN for the treatment of FA-containing sewage.     

盐藻烯醇酶基因表达载体的构建及其转基因烟草的鉴定

以载体pCAMBIA2301为骨架引入pBI121中的GUS基因，构建了简便、实用的中间载体pCAMBIA2301G．将其中一个GUS基因用目的片段盐藻烯醇酶基因替换，构建了可以在植物中高效表达的载体pCAMBIA2301G—enolase，并将其转入根癌农杆菌EHA105中，采用叶盘转化法将盐藻烯醇酶基因转入模式植物烟草中．Southern杂交结果显示，盐藻烯醇酶基因已整合到植物基因组中，在转基因烟草中的拷贝数为2～4个．图6参10     

海洋环境基因组学在海洋生态研究中的应用

海洋环境基因组学是功能基因组学的一个分支学科,它将基因组学的最新技术引入到海洋生态环境的研究当中,得到了许多有价值的研究成果,显示出巨大的发展潜力。文章总结了海洋环境基因组学所开展的工作和已取得的成果,指出目前海洋环境基因组学的工作主要集中在三个领域:海洋生物基因资源文库的建立与完善;海洋生物对环境胁迫的应激反应及生理机制的研究;探索不同生物种群的基因表达谱是否与生物地理分布相关联。并着重讨论了利用基因组学技术研究环境胁迫对海洋生物物种的分布、生物物种间的相互作用以及水生物疾病的影响。如海洋环境基因组学通过测定基因表达谱的变化,研究海洋生物对环境温度的生理响应,绘制出生物地理图谱;通过测定基因转录水平的变化,更深入地了解温度变化所造成的生理支出,从而预测热气候漂移可能给海洋生态环境所造成的影响。在水生物疾病的研究方面,海洋环境基因组学用DNA微阵列技术分析染病生物的基因表达谱,筛选出发生变化的基因表达,为理解病毒感染机制提供了大量信息。还可以深入研究环境胁迫因子与微生物种群群落的组成关系,并由此找到通过维持水体的微生态平衡来消除某些病害发生的环境条件。     

Nitrate reductase for nitrate analysis in water

Nitrate analysis in water is one of the most frequently applied methods in environmental chemistry. Current methods for nitrate are generally based on toxic substances. Here, we show that a viable alternative method is to use the enzyme nitrate reductase. The key to applying this Green Chemistry solution for nitrate analysis is plentiful, inexpensive, analytical grade enzyme. We demonstrate that recombinant Arabidopsis nitrate reductase, expressed in the methylotrophic yeast Pichia pastoris, is a highly effective catalyst for nitrate analysis at 37°C. Recombinant production of enzyme ensures consistent quality and provides means to meet the needs of environmental chemistry.