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.     

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．     

Thermal decomposition of a generation 4.5 PAMAM dendrimer platinum drug conjugate

A nanoscale multivalent platinum drug based on a poly(amidoamine) [PAMAM] dendrimer (generation 4.5, carboxylate surface) has been synthesized and fully characterized using a variety of spectroscopic, chromatographic and thermal methods. Treatment of the dendrimer with an aqueous solution containing an excess diaquo(cis-1,2-diaminocyclohexane)platinum(II) produces a conjugate containing approximately forty (diaminocyclohexane)platinum(II) moieties at the surface of the dendrimer. This material undergoes smooth two-stage thermal decomposition to provide residual platinum oxide reflecting the platinum loading in the drug.     

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。     

The fabrication and characterization of poly(lactic acid) scaffolds for tissue engineering by improved solid–liquid phase separation

In this article a new technique was developed to fabricate scaffolds with a unique microstructure by solid–liquid separation in combination with particulate-leaching. Firstly, the effects of polymer concentration, quenching temperature on the porous morphology and the mechanical property of obtained scaffolds during solid–liquid separation have been investigated. Then, salt granules as porogen were introduced into the solid–liquid phase separation to produce the unique pore structure of the scaffold. The pore diameter of the scaffold could be controlled with the particulate size and the wall of pores possessed special microstructure, which enhanced the pore interconnectivity. The cell culture results confirmed that a good interconnectivity of the scaffold prepared by the improved solid–liquid separation was useful for nutrition transportation and cell proliferation. Copyright © 2003 John Wiley & Sons, Ltd.     

Production of superfine fibers by the electrospinning method

Weakly conducting fluid flows in an electric field in the presence of phase interfaces are investigated theoretically and experimentally. The object of the investigation is an axisymmetric jet. The analysis is carried out within the framework of electrohydrodynamics (EHD) with allowance for surface charge transfer on mobile interfaces. The jet shape is calculated at large distances from the outflow point. The theoretical and experimental data are compared for Newtonian and polymeric fluids.