通过改造工程细胞提高重组蛋白类药物产量的研究进展

哺乳动物细胞是表达蛋白类生物产品的理想宿主细胞。但是,表达水平常较低。通过对工程细胞进行改造,如抑制细胞凋亡、对细胞周期进行调控、提高翻译后加工能力、改变细胞新陈代谢的特性、实现基因组热点整合等,可有效地提高重组蛋白类药物的产量,为生物药品的规模化生产奠定基础     

重组人白介素-2吸入给药对小鼠克雷伯肺炎感染的影响

目的研究重组人白介素-2吸入给药对小鼠克雷伯肺炎感染的保护作用。方法小鼠以滴鼻感染法进行感染。感染过程中,以鼻腔插管注入法和皮下注射给予小鼠重组人白介素-2。同时另设正常组、阴性对照组、模型组和阳性对照组(庆大霉素组)。观察各组动物肺病理变化,比较平均死亡时间和一周内死亡率,测定支气管肺泡洗涤液中总蛋白、白蛋白含量,碱性磷酸酶、乳酸脱氢酶活力。结果重组人白介素-2给药组肺病理变化与模型组和阴性对照组相比显著减轻,支气管肺泡洗涤液中总蛋白、白蛋白含量和碱性磷酸酶活力降低。结论吸入重组人白介素-2缓解感染后肺部病理变化, 为研发重组人白介素-2吸入剂提供了依据。     

Modification of MTT assay conditions to examine the cytotoxic effects of amitraz on the human lymphoblastoid cell line, WIL2NS

Reported parameters of the MTT assay vary widely, and reflect a need to optimise the assay for different cell types. The MTT assay conditions for the human B-lymphocyte-derived cell line WIL2NS were optimised for MTT incubation and formazan development. The optimised MTT assay was validated by examining the effects of the acaride amitraz on WIL2NS. In pH-buffered media in the absence of cells, MTT formed formazan spontaneously, and absorbance was proportional to both the initial concentration of MTT and the time of incubation at 37 °C. One milligram per millilitre MTT was toxic to WIL2NS cells, but the accuracy of the standard curve was reduced when only 0.2 mg/ml MTT was used. Twenty percent SDS in 0.2 M HCl was preferable to DMSO as a solvent for formazan. Exposure to 0.035% amitraz resulted in a significant reduction in WIL2NS cell numbers after only 2 h of exposure. It was concluded that 0.035% of amitraz has the potential to adversely affect lymphocytes in the systemic blood system in humans, and that an optimised MTT assay was obtained by incubating WIL2NS cells with 0.45 mg/ml MTT for 17 h, followed by addition of acidified SDS for 1 h.     

Inhibition of cell cycle and induction of apoptosis by 2‐oxoheksyl isothiocyanate and alyssin in cell lines carrying various inherited BRCA1 mutations

An important aspect of the chemopreventive activity of isothiocyanates (ITC) is their ability to induce cell growth inhibition and apoptosis. In this study, the effect of two sulforaphane analogues, 2‐oxoheksyl isothiocyanate and alyssin, on lymphoblastoid cells, derived from people carrying four different germ‐line mutations in BRCA1 gene, was tested and compared to the effect on wild type cells. The mutations studied were: C61G; 3819del5; 4153delA, and 5382INSC. Changes in cell viability and density after 2‐oxoheksyl isothiocyanate and alyssin treatment were evaluated, as well as cell cycle progression, mitochondrial membrane potential changes, and phosphatidylserine externalization. Both isothiocyanates were shown to reduce cell viability and density in all cell lines tested, as well as the change in cell cycle phase's distribution. The response of cells to two ITC tested was various, as well as mutation type‐modulated. We found that change of cellular maintenance by chemopreventive agents can be modulated by single allele BRCA1 mutation. Drug Dev. Res. 65:84–92, 2005. © 2005 Wiley‐Liss, Inc.     

中国健康人群和甲亢患者细胞色素氧化酶CYP2C19遗传多态性的比较

目的:考察中国健康人群和甲亢患者细胞色素氧化酶CYP2C19遗传多态性的相关性.方法:应用限制性片段长度多态性分析法(PCR-RFLP)对66例甲亢患者和74例健康志愿者进行了基因多态性分析.结果:甲亢患者和健康志愿者中基因型为野生型纯合子wt/wt的分别为34.85%(23/66)和41.89(31/74);基因型为杂合子wt/m1和wt/m2的分别为50.0%(33/66)和45.94%(34/74);基因型为突变型纯合子ml/ml和ml/m2的为15.15%(10/66)和12.16%(9/74).在本实验中未发现m2/m2基因型.甲亢患者和健康志愿受试者的wt/wt和ml/m1的频率没有显著差别.结论:中国甲亢患者和健康人群细胞色素氧化酶CYP2C19遗传多态性在统计学上没有差别.     

Acute toxic effect of the algal yessotoxin on Purkinje cells from the cerebellum of Swiss CD1 mice.

Swiss CD1 mice died less than 2 h after intraperitoneal injection of 420 microg/kg of algal yessotoxin (YTX). The morphological, histochemical and immunocytochemical studies performed on the cerebellar cortex revealed damage to the Purkinje cells. The main cytological alterations were observed in the cytoplasm, while less sufferance was detected in the nucleus. The immunocytochemical experiments showed an increased positivity to S100 protein while there was a decreased response to calbindin D-28K, beta-tubulin and neurofilaments. These changes in intracellular Ca(2+)-binding proteins and the modifications in the cytoskeletal components of Purkinje cells suggest that YTX may be involved in neurological disorders.     

p38 MAPK mediates cardiovascular and behavioral responses induced by central IL-1β and footshock in conscious rats

INTRODUCTION Sharing characteristics of stress mediators[1], cen-tral interleukin-1β (IL-1β) is very important in cardio-vascular and behavioral responses to stressors[2-4]. Itcan induce similar hypertensive responses to stress viacardiovascular regulatory regions[5]. We observed pre-viously that central IL-1β mediated cardiovascular re-sponses to some stressful stimuli, such as conditionedfear stimuli and footshock[6]. Many stressful stimulimay increase IL-1β mRNA expression and …     

茶叶多糖对小鼠激发态免疫功能影响的研究

目的:探讨茶叶多糖对小鼠免疫反应的影响及作用机理.方法:分别用羊红细胞和卵清蛋白为抗原给小鼠注射后,灌服茶叶多糖并设置对照组,检测相应抗体生成水平和血清中IL-2、IFN-γ生成水平.结果:给药组SRBC抗体生成水平和卵清抗体生成水平,IL-2、IFN-γ激发水平均显著高于对照组(P＜0.05).结论:提示茶叶多糖对正常小鼠机体免疫有增强作用.     

Study of effect of oxygen／glucose-deprived culture on the brain-pancreas relative protein in PC12 cells and the mechanism

AIM: To study the effect of oxygen/glucose-deprived (OGD)culture on the expression of a novel protein, brain-pancreas relative protein (BPRP), and the possible regulating mechanism in vitro. BPRP was a key protein found in our previous study of cerebral ischemia. METHODS: PC12 cells was selected and exposed to the Eagle‘s solution containing 1 mmol/L Na2S2O4 for