Expression of glucoamylase gene usingSUC2 promoter inSaccharomyces cerevisiae

Summary The cloning of glucoamylase geneSTA using theSUC2 promoter intoSaccharomyces cerevisiae was performed. The signal sequence ofSTA gene was used for the secretion of glucoamylase protein. The plasmid constructed in this way was named YEpSUCSTA and its expression was identified. The expression of YEpSUCSTA was repressed in the presence of glucose in growth medium, but derepressed when glucose became depleted. YEpSUCSTA showed the similar efficiency of glucoamylase secretion as YEpSTA-F which has the entireSTA gene. Glucoamylase activity in starch-glucose medium was largely increased because cell mass and plasmid stability were high in biosynthesis phase compared to extracellular glucoamylase activities in media which starch or glucose was the only carbon source.     

Reactive oxygen species alter contractile properties of pulmonary arterial smooth muscle

Reactive oxygen species alter pulmonary arterial vascular tone and cause changes in pulmonary vascular resistance. The objective of this investigation was to determine direct effects of oxygen radicals on the contractile properties of pulmonary arterial smooth muscle. Isolated pulmonary arterial rings from Sprague-Dawley rats were placed in tissue baths containing Earle's balanced salt solution (gassed with 95% O2 - 5% CO2, 37 degrees C, pH 7.4). Vessels were contracted with 80 mM KCl to establish maximum active force production (Po). All other responses were normalized as percentages of Po for comparative purposes. Reactive oxygen metabolites were generated enzymatically with either the xanthine oxidase (XO) reaction or the glucose oxidase (GO) reaction, or hydrogen peroxide (H2O2) was added directly to the muscle bath. Exposure to XO, GO, or to H2O2 resulted in a contractile response that was sustained during the 30-min exposure period. The muscle fully relaxed following removal of the reactive oxygen species. Resting tension remained unchanged throughout the experimental period, suggesting no functional change in membrane potential. The contractile response was dose dependent and was not prevented by either cyclooxygenase or lipoxygenase inhibition, or by removal of the endothelium. Pretreatment of vessels with superoxide dismutase (SOD) partially blocked the XO-induced contraction, while mannitol or deferoxamine had no effect on the response to XO. However, pretreatment with catalase (CAT) completely blocked the XO-induced contraction. These data suggest that superoxide ions and hydrogen peroxide are the major causative agents. Following O2-radical exposure, vessels showed a decrease in contractile responsiveness to 80 mM KCl (recovery response), suggesting damage to the smooth muscle cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

福氏痢疾菌膜成分与细菌抗原性和毒力的关系

应用免疫转移技术,用从感染福氏痢疾菌的病人获取的恢复期血清分析了福氏痢疾菌膜成分与细菌抗原性和毒力的关一。发现福氏痢疾菌的膜蛋白67kD和63kD均含有两种成分,一种和膜蛋白60kD都可能是保护性抗原,而另一种与膜蛋白78kD和35kD一样与福氏痢疾菌的毒力有关。采用微细胞同位素掺入示踪显示这些与病人恢复期血清反应的膜蛋白由质粒编码。     

Entrapment of microbial cells and organelles with hydrophilic urethane prepolymers

Summary Microbial cells and cellular organelles were immobilized by mixing aqueous suspensions of the biocatalysts with water-miscible urethane prepolymers. Thus immobilized preparations of acetone-dried cells of Arthrobacter simplex and thawed cells of Nocardia rhodocrous showed appreciable {ie351-1} activities in the transformation of hydrocortisone into prednisolone and 4-androstene-3,17-dione to androst-1,4-diene-3,17-dione, respectively. The activities of catalase and alcohol oxidase were observed in the immobilized peroxisomes (microbodies) of a methanol-grown yeast Kloeckera sp. No. 2201. Yeast mitochondria entrapped with the prepolymer showed adenylate kinase activity. These results indicate the usefulness of the urethane prepolymers as convenient materials for entrapment of not only enzymes, but also organelles and microbial cells.     

高产青霉素酰化酶巨大芽胞杆菌的诱变选育及产酶条件优化

【目的】选育高产青霉素G酰化酶(PGA)工业菌株。【方法】采用LiCl-紫外线复合诱变以及常压室温等离子体(ARTP)诱变技术对巨大芽胞杆菌(Bacillus megaterium) ATCC 14945进行处理。处理菌体涂平板后,将长出的菌落接种到液体培养基中,向培养6 h后的二代菌液中添加终浓度为0.1%的苯乙酸,28 °C、250 r/min条件下诱导培养40 h。对离心后获得的上清(粗酶液)采用NIPAB法测定PGA酶活力。以PGA酶活力最高的菌株为材料,对苯乙酸最佳添加量和最佳诱导时间进行优化,采用NIPAB法测定PGA酶活力。采用SDS-PAGE检测诱变前后巨大芽胞杆菌粗酶液中PGA的蛋白特性。【结果】从诱变菌落中筛选到PGA酶活力为39.60 U/mL的菌株12-4,酶活力比出发菌株提高了8.5倍。该菌株在液体培养6 h后添加终浓度为0.2%的苯乙酸,继续培养50 h后,PGA酶活力可达78.45 U/mL,比出发菌株提高了16.8倍。诱变前后菌株培养液中的PGA蛋白均具α、β亚基;诱变后菌株PGA α亚基的量没有明显变化,β亚基的量明显增多;α、β亚基之间的蛋白条带明显增多。【结论】采用诱变技术可提高巨大芽胞杆菌PGA活性,获得的诱变菌株12-4及培养条件对PGA工业化生产具有重要价值。     

非生物胁迫下耐辐射异常球菌drB0118基因功能分析

【目的】鉴定和确定被预测为编码干燥相关蛋白的耐辐射异常球菌(Deinococcus radiodurans) drB0118基因功能,探讨该基因对盐、渗透和氧化胁迫抗性的作用。【方法】构建drB0118基因缺失突变株(ΔB0118),通过氯化钠、D-山梨糖醇和过氧化氢等胁迫冲击实验及氧化胁迫条件下qRT-PCR分析,研究drB0118突变对非生物胁迫反应及氧化胁迫相关基因表达的影响。【结果】drB0118突变导致菌株对NaCl和D-sorbitol胁迫的抗性降低;对氧化胁迫(H2O2)敏感;qRT-PCR分析显示,drB0118突变引起氧化胁迫抗性基因pod和oxyR分别下调4倍和10倍。【结论】D. radiodurans中drB0118参与了盐、渗透和氧化等多种非生物胁迫反应。     

Economic burden of dengue fever in China: A retrospective research study

BackgroundDengue fever has been a significant public health challenge in China. This will be particularly important in the context of global warming, frequent international travels, and urbanization with increasing city size and population movement. In order to design relevant prevention and control strategies and allocate health resources reasonably, this study evaluated the economic burden of dengue fever in China in 2019.MethodsThe economic burden of dengue fever patients was calculated from both family and the organisation perspectives. A survey was conducted among 1,027 dengue fever patients in Zhejiang, Chongqing, and Yunnan Provinces. Treatment expenses, lost working days, and insurance reimbursement expenses information were collected to estimate the total economic burden of dengue fever patients in 2019. The expenditures related to dengue fever prevention and control from government, Center for Disease Control and Prevention (CDC), communities and subdistrict offices of 30 counties (or districts) in Zhejiang Province and Chongqing City were also collected.ResultsThe direct, indirect and total economic burden for dengue fever patients in 2019 in the three Provinces were about 36,927,380.00 Chinese Yuan (CNY), 10,579,572.00 CNY and 46,805,064.00 CNY, respectively. The costs for prevention and control of dengue fever for the counties (or districts) without cases, counties (or districts) with imported cases, and counties (or districts) with local cases are 205,800.00 CNY, 731,180.00 CNY and 6,934,378.00 CNY, respectively. The total investment of dengue fever prevention and control in the 30 counties in China in 2019 was approximately 3,166,660,240.00 CNY.ConclusionThe economic burden of dengue fever patients is relatively high, and medical insurance coverage should be increased to lighten patients’ direct medical economic burden. At the same time, the results suggests that China should increase funding for primary health service institutions to prevent dengue fever transmission.     

天山林区六种灌木生物量的建模及其器官分配的适应性

灌木全株生物量估算模型的构建仍存在一定困难,对灌木生物量在器官分配上所体现的适应性研究也不够充分。以天山林区6种常见灌木为研究对象,在天山的东段、中段、西段林区分别设置样地进行群落调查,由此以全株收获法取得6种常见灌木若干标准株的全株、根、枝、叶及各径级根的生物量,将D~2H(地径平方与高度的乘积)与V(冠幅面积与高度的乘积)分别选为估测模型的自变量,通过回归分析法建立了各种灌木全株生物量的最优估算模型,然后比较了此6种灌木全株生物量在营养器官上分配差异以及根系生物量在径级上的分配差异。结果表明:(1)天山林区6种常见灌木中,小檗(Berberis heteropoda Schrenk)、忍冬(Lonicera hispida Pall.ex Roem.et Schuet.)、栒子(Cotoneaster melanocarpus Lodd.)的全株生物量约为8.48—9.01 kg,蔷薇(Rosa spinosissima L.)、绣线菊(Spiraea hypericifolia L.)、方枝柏(Juniperus pseudosabina Fisch.et Mey.)的全株生物量约为2.71—3.20 kg;(2)蔷薇、绣线菊、栒子的全株生物量最优估测模型是以V为自变量的函数,小檗、忍冬、方枝柏的全株生物量最优估测模型是以D~2H为自变量的函数,各模型R~2值均在0.850以上,且在P0.05水平上达到显著,模型模拟结果达到了较高的准确度;(3)6种灌木全株生物量在根、枝上的分配比重差异不显著,仅在叶上的分配比重有差异(P0.05);根系生物量在径级上的分配均呈现随根系径级下降而减少的规律,6种灌木在径级大于2 mm根上的分配比重存在差异(P0.05,径级大于20 mm根为P0.01水平);(4)6种灌木全株生物量在营养器官上的分配差异以及根系生物量在径级上的分配差异均体现了各物种对其生境选择的适应策略。