乙醇氧化酶两点速率法测定血清微量乙醇

目的：建立一个灵敏、准确、特异、试剂稳定的乙醇氧化酶法用于测定血清微量乙醇浓度。方法：利用乙醇氧化酶与过氧化物酶偶联，通过优化反应体系及自动生化分析仪分析参数，采用两点速率法对乙醇进行分析。结果：乙醇浓度在2000mg／L以下线性良好；批内RSD小于2．81％，批间RSD小于4．51％；平均回收率为100．7％；本法（Y）与气相色谱法（X）比较具有良好的相关性，Y=0．996X＋3．58，r=0．981。胆红素159．1μmol／L、甘油三酯14．31mmol／L、血红蛋白5．8g／L以下对本法测定乙醇结果无干扰。结论：建立的两点速率法测定乙醇灵敏度高、准确度、特异性均较好，特别适合于血清乙醇浓度处于轻度超标的疑似酒后者的标本检测。     

蛋白酶体功能障碍对多巴胺能PC12细胞的特异性损伤

目的探讨蛋白酶体抑制剂lactacystin对多巴胺能PC12细胞的特异性损伤。方法不同浓度的lactacystin(1、5、10、15和20μmol/L)分别处理多巴胺能PC12细胞和胶质瘤U251细胞24 h,MTT法检测细胞活力;50μmol/L的lactacystin处理U251细胞24 h,MTT法检测细胞活力;10、20μmol/L lactacystin处理PC12细胞,W estern B lot检测细胞内多泛素化蛋白含量;单胺氧化酶B抑制剂selegiline(500μmol/L)和特异性酪氨酸羟化酶抑制剂-αMT(1 mmol/L)提前4 h预处理PC12细胞,再与10μmol/Llactacystin共同作用24 h,MTT法检测细胞活力,W estern B lot检测多泛素化蛋白含量。结果Lactacystin呈剂量依赖性损伤多巴胺能PC12细胞,其对胶质瘤U251细胞无毒性作用,而且其毒性与细胞内多泛素化蛋白生成增加相关。用以增加细胞内多巴胺含量的selegiline和用以减少细胞内多巴胺含量的α-MT都导致lactacystin毒性增强。结论蛋白酶体功能障碍特异性损伤多巴胺能细胞,而其特征性的神经递质多巴胺在其易感性中的作用复杂。     

二茂铁介导的酶电极血糖仪的研制

以二茂铁为电子媒介体,研究制作了一次性干法酶电极的便携式血糖仪。血糖检测只需1．5μl的血液标本,并在20s 就能得出检测结果,检测的范围为50-500mg/dl,精度为±1 mg/dl。血糖仪体积为94．3mm×49．2mm×17．8 mm,操作简单, 便于携带。其性能与通过美国FDA认证的YSI 2300 Stat Plus葡萄糖分析仪进行比较研究。结果表明,检测结果的变异系数为3．0%-7．8％,具有线性关系(y=1．006x-1．37,r=0．990,n=118)和很好的可比性,误差图格分析显示所有的数据都在可接受的A、B区。引入二茂铁后,血糖仪的灵敏度显著提高,是一个有效的用于血糖分析的体外诊断仪器。     

海洛因对大鼠黄嘌呤氧化酶和血尿酸的影响

目的:检测黄嘌呤氧化酶(xanthineoxidase ,XOD)和血浆尿酸含量,研究海洛因给药对嘌呤核苷酸分解代谢的影响。方法:建立海洛因给药、停药大鼠模型,测定尿酸及XOD含量。结果:给药组尿酸及XOD含量高于对照组。与给药组比较,两个停药组尿酸含量、血浆和肝XOD含量降低;停药8d组脑顶叶XOD含量降低,脑干XOD含量略有下降趋势。但两个停药组脑干XOD含量仍高于对照组。结论:海洛因通过增加XOD的含量,使嘌呤核苷酸分解代谢增强,且脑中这一作用消除较慢。     

Methylamine but not mafenide mimics insulin-like activity of the semicarbazide-sensitive amine oxidase-substrate benzylamine on glucose tolerance and on human adipocyte metabolism

It has been reported that benzylamine reduces blood glucose in rabbits, stimulates hexose uptake, and inhibits lipolysis in mouse, rabbit, and human adipocytes. In the presence of vanadate, benzylamine is also able to improve glucose disposal in normoglycaemic and diabetic rats. Such insulin-mimicking properties are the consequence of hydrogen peroxide production during benzylamine oxidation by semicarbazide-sensitive amine oxidase (SSAO). The aim of the study was to determine whether other SSAO-substrates could share such potential antidiabetic properties. Thus, mafenide, a synthetic antimicrobial sulfonamide structurally related to benzylamine, and which has been recently reported to interact with SSAO, was tested in the above mentioned models, in parallel with methylamine, a proposed endogenous SSAO-substrate. All tested amines stimulated glucose uptake and inhibited lipolysis in rat and mouse fat cells. Methylamine and benzylamine, but not mafenide, reduced the hyperglycaemic response during a glucose tolerance test in rabbits while the three amines tested were devoid of insulin-releasing activity under both in vivo and in vitro conditions. In human adipocytes, mafenide did not stimulate glucose transport since it was not a high-affinity substrate for SSAO and generated less hydrogen peroxide than benzylamine or methylamine. Therefore, mafenide could not be considered as an antidiabetic drug despite being oxidized and exhibiting insulin-mimicking effects in rat and mouse adipocytes. By contrast, the endogenous substrate methylamine improved glucose utilization in all in vitro and in vivo models, leading to consider novel SSAO substrates as drugs with potential anti-hyperglycaemic properties.     

Monoamine oxidase inhibitory activities of the scorpion Mesobuthus gibbosus (Buthidae) venom peptides.

In the present study, crude venom of Mesobuthus gibbosus (Buthidae), a scorpion distributed all over Anatolia was isolated and purified by the Sephadex G-50 gel filtration and high pressure liquid chromatographic (HPLC) separation. Two of the five fractions (fractions 4 and 5) obtained from the Sephadex G-50 filtration and detected as lethal on mice and Musca domestica larvae in in vivo toxicity tests, were independently subjected to the HPLC separation. Only one of seven fractions (fraction 5.5*) obtained from the HPLC separation of the fraction 5 was found to be extremely lethal. Sodium dodecylsulfate polyacrylamide gel electrophoretic (SDS-PAGE) analysis of the crude venom and its chromatographic fractions demonstrated that crude venom consisted of peptides with molecular weights of 6500-210,000 Da. The neurotoxic fraction 5.5* appeared as a single band of 28,000 Da and two bands of 6200 and 22,000 Da in SDS-PAGE under non-reducing and reducing conditions, respectively, suggesting that it might consist of two chains attached by a disulfide bridge. Fractions 5 and 5.5* inhibited monoamine oxidase A (MAO-A) of rat liver reversibly and non-competitively, in a concentration-dependent manner. Fraction 5.5* appeared as a potent and specific MAO-A inhibitor with a Ki value of 0.12 mg venom proteinml(-1). The inhibitory effect of venom peptide 5.5* on MAO-A was found to be dependent on the preincubation time suggesting that the peptide binds to some site other than the substrate-binding site. Results of the present study demonstrated that M. gibbosus venom contains a peptide with specific MAO-A inhibitory activity which may be responsible for the anxiogenic effects of the scorpion venoms on animals and humans.     

线粒体与吗啡诱导的肝损伤作用

我们用药掺食法培养吗啡依赖大鼠模型，研究了吗啡依赖大鼠肝线粒体自由基损伤及其结构与功能的变化。结果发现：超氧化物歧化酶（ＳＯＤ）活性明显降低、丙二醛（ＭＤＡ）含量明显升高；细胞色素Ｃ氧化酶活性、心磷脂含量和膜流动性都明显降低。这表明吗啡依赖大鼠肝线粒体自由基清除能力降低，导致自由基损伤增加，并进一步影响到线粒体的结构与功能。线粒体结构与功能的这种改变可能在吗啡诱导的肝损伤中有重要作用。     

伴生菌对猪苓几种酶活性的影响

目的 :研究猪苓Grifolaumbellata在与伴生菌Companionfungus共培养过程中 ,伴生菌对猪苓几种酶的活性影响。方法 :测定与伴生菌共培养过程中猪苓菌丝几丁质酶、β-1,3-葡聚糖酶、蛋白酶及胞外多酚氧化酶的活性。结果 :伴生菌能诱导猪苓几丁质酶及 β-1,3-葡聚糖酶活性的提高 ;蛋白酶活性没有明显变化 ;液体培养时 ,在培养的后期猪苓与伴生菌共培养的发酵液中两种胞外多酚酶活性均居于猪苓与伴生菌单独培养的酶活性之间。结论 :伴生菌与猪苓的营养互补可能为猪苓提供一些菌核形成的相关物质 ,从而有利于猪苓形成菌核。     

Cancer chemoprevention by tea polyphenols through modulating signal transduction pathways

The action mechanisms of several chemopreventive agents derived from herbal medicine and edible plants have become attractive issues in cancer research. Tea is the most widely consumed beverage worldwide. Recently, the cancer chemopreventive actions of tea have been intensively investigated. It have been demonstrated that the active principles of tea were attributed to their tea polyphenols. Recently, tremendous progress has been made in elucidating the molecular mechanisms of cancer chemoprevention by tea and tea polyphenols. The suppression of various tumor biomarkers including growth factor receptor tyrosine kinases, cytokine receptor kinases, PI3K, phosphatases, ras, raf, MAPK cascades, N x FB, I x B kinase, PKA, PKB, PKC, c-jun, c-fos, c-myc, cdks, cyclins, and related transducing proteins by tea polyphenols has been studied in our laboratory and others. The I x B kinase (IKK) activity in LPS-activated murine macrophages (RAW 264.7 cells) was found to be inhibited by various tea polyphenols including (-) epigallocatechin-3-gallate (EGCG), theaflavin (TF-1), theaflavin-3-gallate (TF-2) and theaflavin-3,3'-digallate (TF-3). TF-3 inhibited IKK activity in activated macrophages more strongly than did the other tea polyphenols. TF-3 inhibited both IKK1 and IKK2 activity and prevented the degradation of I x B x and I x B x in activated macrophage cells. The results suggested that the inhibition of IKK activity by TF-3 and other tea polyphenols could occur by a direct effect on IKKs or on upstream events in the signal transduction pathway. TF-3 and other tea polyphenols blocked phosphorylation of IB from the cytosolic fraction, inhibited NFB activity and inhibited increases in inducible nitric oxide synthase levels in activated macrophage. TF-3 and other tea polyphenols also inhibited strongly the activities of xanthine oxidase, cyclooxygenase, EGF-receptor tyrosine kinase and protein kinase C. These results suggest that TF-3 and other tea polyphenols may exert their cancer chemoprevention through suppressing tumor promotion and inflammation by blocking signal transduction. The mechanisms of this inhibition may be due to the blockade of the mitogenic and differentiating signals through modulating EGFR function, MAPK cascades, NFkappaB activation as well as c-myc, c-jun and c-fos expression.     

κ-硒化角叉菜胶对大鼠红细胞膜流动性与封闭度的影响(英文)

通过测定大鼠红细胞膜荧光偏振度P和N A DH-细胞色素c氧化还原酶的变化,研究kappa-硒化卡拉胶对红细胞膜流动性和封闭度的影响。结果表明,kappa-硒化卡拉胶140mg·kg~(-1)·d~(-1)×30 d可显著降低大鼠红细胞膜荧光偏振度,即可显著提高细胞膜流动性(P<0.05),ig 140及70 mg·kg~(-1)·g~(-1)×30d可显著提高大鼠红细胞膜封闭度(P<0.05)。