Simultaneous quantification of leukotrienes and hydroxyeicosatetraenoic acids in cell culture medium using liquid chromatography/tandem mass spectrometry

Leukotrienes (LTs) and hydroxyeicosatetraenoic acids (HETEs) are important bioactive lipid mediators that participate in various pathophysiological processes. To advance understanding of the mechanisms that regulate these mediators in physiological and pathological processes, an analytical method using liquid chromatography/tandem mass spectrometry for the simultaneous quantification of LTB₄, LTC₄, LTD₄, LTE₄, 5‐HETE_, 8‐HETE, 12‐HETE and 15‐HETE in cell culture media was developed. A Supel?‐Select HLB solid‐phase extraction cartridge was used for sample preparation. The compounds were separated on a C₁₈ column using gradient elution with acetonitrile–water–formic acid (20:80:0.1, v/v/v) and acetonitrile–formic acid (100:0.1, v/v). The calibration curves of LTB₄, LTD₄, LTE₄ and HETEs were linear in the range of 0.025–10 ng/mL, and the calibration curve of LTC₄ was linear in the range of 0.25–10 ng/mL. Validation assessment showed that the method was highly reliable with good accuracy and precision. The stability of LTs and HETEs was also investigated. Using the developed method, we measured LTs and HETEs in the culture supernatant of the human mast cell line HMC‐1. The present method could facilitate investigations of the mechanisms that regulate the production, release and signaling of LTs and HETEs. Copyright © 2014 John Wiley & Sons, Ltd.     

我国上市公司资本结构影响因素实证分析

本文选取了可能影响企业资本结构的多个指标变量进行分析 ,利用主成分分析提供的方法将变量综合成彼此互不相关的少数几个主成分。再用主成分 (作为回归自变量 )对企业的 5种资本负债比 (作为回归因变量 )进行多元回归分析 ,得出了影响企业资本结构的主要因素 ,以及这些因素与企业资本结构之间的关系 ,为企业确定资本结构提供参考依据 ,为企业财务决策提供支持。     

我国上市公司资本结构决策的实证研究

本在已有资本结构理论研究成果的基础上，对30家沪市上市公司的资本结构进行了实证研究，提出了在我国资本市场环境下优化企业资本结构的对策建议。     

上市公司资本成本面板数据模型与实证分析

本文利用我国上市公司1997至2002年的有关财务数据,建立了我国上市公司资本成本面板数据模型,运用该模型分析了我国上市公司资本成本与财务杠杆、企业规模的关系.结果表明,财务杠杆的提升、企业规模的扩大将使资本成本下降.该结论对我国上市公司优化资本结构、提高企业价值具有一定的参考价值.     

乳糖修饰壳聚糖微载体的制备及对培养肝细胞代谢活性的影响

液体石蜡作分散介质。戊二醛作交联刑，通过反相悬浮聚合制备了微米级的壳聚糖微载体．环氧氟丙烷活化后，乳糖进行修饰．用孔糖修饰的微载体进行原代大鼠肝细胞培养，利用相差显微镜对培养细胞进行形态观察，并测定肝细胞的代谢活性，结果显示，乳糖修饰壳聚糖微载体是一种优良的肝细胞培养支架．     

创业投资与金融体系关系的实证研究

本文实证研究了创业投资产业与金融体系的内在关系,并对目前我国以银行为中心的金融体系下发展创业投资产业提出了一些建议.     

Altered protein synthesis in rat kidney cells exposed to semiconductor materials

It is thought that the extensive industrial use of arsenic, gallium and indium, which have applications as the materials for III–V semiconductors, will increase human exposure to these compounds in the near future. We have undertaken the development of new biological indicators for assessing exposure to these elements. Element-specific alterations in protein synthesis patterns were expected to occur following exposure to arsenic compounds. We examined alterations in protein synthesis in primary cultures of rat kidney proximal tubule epithelial cells by sodium arsenite, gallium chloride and indium chloride, utilizing two-dimensional gel electrophoresis. After incubation with the chemicals for 20 h, newly synthesized proteins were labeled with [³⁵S]methionine. A protein with a molecular weight (M_r) of 30 000 was markedly induced on exposure to 10 μM arsenite or 300 μM gallium chloride, and synthesis of proteins with M_r values of 85 000, 71 000, 65 000, 51 000, 38 000 and 28 000 were also increased by exposure to arsenite and gallium chloride. No significant changes were observed upon exposure to indium. Some of these increased proteins could be heat-shock proteins.     

Uptake and excretion of total inorganic arsenic by the freshwater alga Chlorella vulgaris

Arsenic-tolerant freshwater alga Chlorella vulgaris which had been collected from an arsenicpolluted environment were tested for uptake and excretion of inorganic arsenic. Approximately half the quantity of arsenic taken up by C. vulgaris was estimated to be adhered to the extraneous coat (10 wt %) of the cell. The remainder was bioaccumulated by the cell. Both adhered and accumulated arsenic concentrations increased with an increase in arsenic(V) concentration of the aqueous phase. Arsenic(V) accumulation was affected by the growth phse: arsenic was most actively accumulated when the cell was exposed to arsenic during the early exponential phase and then accumulation decreased with an increase in culture time exposed to arsenic. The alga grew well in the modified Detmer (MD) medium containing 1 mg As(III) dm^?3 and the growth curve was approximated by a ‘logistic equation’. Arsenic(III) was accumulated up to the second day of the culture time and arsenic(III) accumulation decreased with an increase in the culture time after that. Arsenic accumulation was also largely affected by various nutrients, especially by managanese, iron and phosphorus compounds. A modified MD medium with the three nutrients was proposed for the purpose of effective removal of arsenic from the aqueous phase. Using radioactive arsenate (Na₂H⁷⁴AsO₄), the arsenic accumulated was found to be readily excreted under conditions which were unfavourable for the multiplication of C. vulgaris.     

人原始生殖细胞的分离和体外培养

从4～10周龄药物流产胚胎的生殖嵴和肠系膜组织中分离原始生殖细胞(primordial germ cells,PGCs),培养在添加人重组白血病抑制因子(rh LIF)、人重组碱性成纤维细胞生长因子(rh bFGF)和Forskolin的小鼠饲养层细胞上.经过4～7 d培养,PGCs形成典型的鸟巢状集落.集落在传代过程中保持碱性磷酸酶活性,且胚胎阶段性特异抗原1(SSEA-1)、胚胎阶段性特异抗原3(SSEA-3)免疫荧光染色呈阳性.具有分化潜能的PGCs能在体外连续传代培养12代.结果表明从药物流产胚胎中分离的人类PGCs可以在体外培养成为具有分化潜能的多能性干细胞.     

槲寄生蛋白注射液细胞毒活性测定的MTT方法探讨

采用MTT比色法对体外培养的肿瘤细胞进行细胞毒作用实验，验证槲寄生蛋白注射液体外抗肿瘤效果．并且鉴定这种检测方法的有效性。实验结果表明槲寄生蛋白注射液有一定的体外抗肿瘤效果；MTT比色法是一种可用的体外细胞毒作用检测法，为新药品的开发提供了实验依据。