影响正常口腔念珠菌检出率的方法学研究

目的:研究不同检测方法对正常口腔念珠菌检出率的影响,寻找一种比较简便可靠的检测方法.方法:以健康的平均年龄7.4岁的儿童为检测人群,比较不同的取样部位,取样方法,检测方法,被检人群口腔中白色念珠菌以及其他念珠菌的检出率.结果:取样和检测方法对检出率有不同程度的影响,PCR检测方法的检出率显著高于培养法.结论:黏膜拭子加离心,和CHROMagar CandidaTM鉴定培养基相结合的方法是一种简便理想的分离培养方法,PCR方法则敏感度更高.     

Evaluation of oxygen adaptation and identification of functional bacteria composition for anammox consortium in non-woven biological rotating contactor

In this study, the anammox consortium was found to adapt to the wastewater containing dissolved oxygen (DO), as the DO was gradually increased. Batch tests indicated the maximum aerobic ammonium oxidizing activity of the consortium was 1.38mmolNH(4)(+)-N(gVSS)(-1)day(-1), which played key roles in the oxygen consumption process; the maximum anaerobic ammonium oxidizing activity was slightly decreased after long-term oxygen exposure, but only from 21.23mmolNH(4)(+)-N(gVSS)(-1)day(-1) to 20.23mmolNH(4)(+)-N(gVSS)(-1)day(-1). Microbiological community analysis identified two strains similar to Nitrosomonas eutropha were responsible for oxygen consumption, which were able to exist in the autotrophic anaerobic condition for long periods and protect anammox bacteria Planctomycetales from the influence of oxygen. Microbiological composition analysis showed Nitrosomonas and Planctomycetales approximately accounted for 10% and 70% of the bacteria, respectively. The possibility of cultivation anammox consortium in presence of DO will lead to substantial savings of energy and resource in the industrial application.     

口蹄疫病毒3A基因在大肠杆菌中的高效表达

将口蹄疫病毒(Foot\|and\|Mouth Disease Virus,FMDV)的3A基因克隆到线形化的原核表达载体pProEX\|HTb中,转化大肠杆菌BL21和DH5α,经氨苄抗性筛选得到阳性克隆,IPTG诱导表达。SDSPAGE和West bolt结果证实大肠杆菌菌体不可溶性蛋白中富含3A蛋白,说明3A蛋白在表达产物中以包涵体的形式存在,所表达的蛋白含量占菌体蛋白的29.2%。     

Pyrosequencing Reveals Contrasting Soil Bacterial Diversity and Community Structure of Two Main Winter Wheat Cropping Systems in China

Microbes are key components of the soil environment, playing an important role in maintaining soil health, sustainability, and productivity. The composition and structure of soil bacterial communities were examined in winter wheat–rice (WR) and winter wheat–maize (WM) cropping systems derived from five locations in the Low-Middle Yangtze River plain and the Huang-Huai-Hai plain by pyrosequencing of the 16S ribosomal RNA gene amplicons. A total of 102,367 high quality sequences were used for multivariate statistical analysis and to test for correlation between community structure and environmental variables such as crop rotations, soil properties, and locations. The most abundant phyla across all soil samples were Proteobacteria, Acidobacteria, and Bacteroidetes. Similar patterns of bacterial diversity and community structure were observed within the same cropping systems, and a higher relative abundance of anaerobic bacteria was found in WR compared to WM cropping systems. Variance partitioning analysis revealed complex relationships between bacterial community and environmental variables. The effect of crop rotations was low but significant, and interactions among soil properties, locations, and crop rotations accounted for most of the explained variation in the structure of bacterial communities. Soil properties such as pH, available P, and available K showed higher correlations (positive or negative) with the majority of the abundant taxa. Bacterial diversity (the Shannon index) and richness (Chao1 and ACE) were higher under WR than WM cropping systems.     

不同批号蕲蛇酶在家兔血液凝固系统上的生物活性检测

目的：研究不同批号蕲蛇酶静脉给予家兔后,对血液凝固系统各项指标的影响,以判定其药效的稳定性。方法：取家兔,纯白色,体重1．5－2．5kg,雌雄兼用,每日静脉注入蕲蛇酶1．0u/kg.d^-1,连用3－5天,药前、药后从心脏取血6ml,取全血1ml作血栓形成;余血以3．0％枸橼酸钠按1：9抗凝,分离富血小板血浆（PRP）,测定其血小板数目和血小板聚集率,以贫血小板血浆（PPP）测定凝血酶时间（TT）,凝血酶原时间（PT）,部分凝血活酶时间（KPTT）以及纤维蛋白原（Fg)含量。结果：5个批号的蕲蛇酶药后3天或5天均可明显使血栓形成的重量减轻,长度缩短,Fg含量减少（P＜0．01）,TT,PT,KPTT均延长（P＜0．05－0．01）,血小板数轻度减少,聚集率抑制达20％－50％,结论：不同批号的蕲蛇酶静脉给予家兔,具有显著的作用于血液凝固系统,导致血栓形成减少,表明对心脑血管内血栓形成药学相同,说明用家 兔作为鉴定药品质量是可行而必须。     

Identification and characterization of a novel salt‐tolerant esterase from a Tibetan glacier metagenomic library

A salt‐tolerant esterase, designated H9Est, was identified from a metagenomic library of the Karuola glacier. H9Est gene comprised 1071 bp and encoded a polypeptide of 357 amino acids with a molecular mass of 40 kDa. Sequence analysis revealed that H9Est belonged to the family IV of bacterial lypolitic enzyme. H9Est was overexpressed in Escherichia coli and the purified enzyme showed hydrolytic activity towards p‐nitrophenyl esters with carbon chain from 2 to 8. The optimal esterase activity was at 40°C and pH 8.0 and the enzyme retained its activity towards some miscible organic solvents such as polyethylene glycol. A three‐dimensional model of H9Est revealed that S200, D294, and H324 formed the H9Est catalytic triad. Circular Dichroism spectra and molecular dynamic simulation indicated that the esterase had a wide denaturation temperature range and flexible loops that would be beneficial for H9Est performance at low temperatures while retaining heat‐resistant features. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:890–899, 2015     

胡杨甜菜碱醛脱氢酶基因的功能分化

甜菜碱醛脱氢酶(BADH)在植物抗逆反应中发挥着重要作用。文中从胡杨cDNA克隆到2个甜菜碱醛脱氢酶基因,分别命名为PeBADH1和PeBADH2。PeBADH1和PeBADH2均编码503个氨基酸的蛋白质,预测分子量分别是54.93 kDa和54.90 kDa。组织表达模式分析发现这2个基因在正常生长、盐和H2O2胁迫下,在不同组织中的表达模式有较大差异。在大肠杆菌中表达并纯化了2个基因的重组蛋白。酶活性分析显示PeBADH1和PeBADH2蛋白对底物的活性分别是0.073μmol/(min.mg)和0.107μmol/(min.mg)。热力学稳定性分析显示这2个蛋白的热力学稳定性具有明显差异。因此,基因表达模式差异与蛋白质酶学性质的不同预示着这2个基因可能存在功能上的分化。     

沙坡头人工固沙区的种子库动态

沙坡头人工固沙区的土壤种子库中共有４６种植物的种子,属于１７个科。其中菊科植物最多,达１１种;其次为藜科,７种;禾本科７种;豆科４种。既是现有植被组分种,其种子又存在于种子库中的,占总种数的３３．３％ 。油蒿（Ａｒｔｅｍ ｉｓｉａ ｏｒｄｏｓｉｃａ）、花棒（Ｈｅｄｙｓａｒｕｍ ｓｃｏｐａｒｉｕｍ ）和柠条（Ｃａｒａｇａｎａ ｋｏｒｓｈｉｎｓｋｉｉ）的种子产量为５４５±６７６．７３粒／ｍ ２、６．６７±１３．７４粒／ｍ ２、０．０３２±２．４３×１０－ ３粒／ｍ ２。油蒿在植株上成熟时不受虫害,而花棒、柠条的种子受害比例分别为３１．９２％ 和８３．２９％ 。在最适埋藏深度下,油蒿、柠条、花棒种子的累积发芽率分别可达７８．３％ 、８５．０％ 和６６．７％ ;萌发速度：油蒿＞ 柠条＞ 花棒。根据种子库动态分析和植被调查结果可初步预测：沙坡头人工固沙植被的演替趋向将是灌木以油蒿为主,草本以雾冰藜（Ｂａｓｓｉａ ｄａｓｙｐｈｙｌｌａ）、小画眉草（Ｅｒａｇｒｏｓｔｉｓｐｏａｅｏｉｄｅｓ）为主的群落     

Inhibition of mitochondrial complex I improves glucose metabolism independently of AMPK activation

Accumulating evidences showed metformin and berberine, well‐known glucose‐lowering agents, were able to inhibit mitochondrial electron transport chain at complex I. In this study, we aimed to explore the antihyperglycaemic effect of complex I inhibition. Rotenone, amobarbital and gene silence of NDUFA13 were used to inhibit complex I. Intraperitoneal glucose tolerance test and insulin tolerance test were performed in db/db mice. Lactate release and glucose consumption were measured to investigate glucose metabolism in HepG2 hepatocytes and C2C12 myotubes. Glucose output was measured in primary hepatocytes. Compound C and adenoviruses expressing dominant negative AMP‐activated protein kinase (AMPK) α1/2 were exploited to inactivate AMPK pathway. Cellular NAD⁺/NADH ratio was assayed to evaluate energy transforming and redox state. Rotenone ameliorated hyperglycaemia and insulin resistance in db/db mice. It induced glucose consumption and glycolysis and reduced hepatic glucose output. Rotenone also activated AMPK. Furthermore, it remained effective with AMPK inactivation. The enhanced glycolysis and repressed gluconeogenesis correlated with a reduction in cellular NAD⁺/NADH ratio, which resulted from complex I suppression. Amobarbital, another representative complex I inhibitor, stimulated glucose consumption and decreased hepatic glucose output in vitro, too. Similar changes were observed while expression of NDUFA13, a subunit of complex I, was knocked down with gene silencing. These findings reveal mitochondrial complex I emerges as a key drug target for diabetes treatment. Inhibition of complex I improves glucose homoeostasis via non‐AMPK pathway, which may relate to the suppression of the cellular NAD⁺/NADH ratio.