Characterization of prolidase activity using capillary electrophoresis with tris(2,2'-bipyridyl)ruthenium(II) electrochemiluminescence detection and application to evaluate collagen degradation in diabetes mellitus

A new method for prolidase (PLD, EC 3.4.13.9) activity assay was developed based on the determination of proline produced from enzymatic reaction through capillary electrophoresis (CE) with tris(2,2'-bipyridyl)ruthenium(11) [Ru(bpy)(3)(2+)] electrochemiluminescence detection (ECL). A detection limit of 12.2 fmol (S/N = 3) for proline, corresponding to 1.22 x 10(-8) units of prolidase catalyzing for 1 min was achieved. PLD activity determined by CE-ECL method was in agreement with that obtained from the classical Chinard's one. CE-ECL showed its powerful resolving ability and selectivity as no sample pretreatmentwas needed and no interference existed. The clinical utility of this method was successfully demonstrated by its application to assay PLD activity in the serum of diabetic patients in order to evaluate collagen degradation in diabetes mellitus (DM). The results indicated that enhanced collagen degradation occurred in DM.

富铬酵母对实验性糖尿病小鼠模型的治疗

目的：三价铬作为葡萄糖耐量因子的有效活性成分，具有改善糖尿病人的糖代谢和脂代谢的作用，因此补充三价铬是糖尿病治疗中的有效的营养措施，富铬酵母是目前向人体提供三价铬的有效途径，并且葡萄糖耐量因子可以提高靶组织对胰岛素的敏感性而不促进胰腺的胰岛素分泌，为治疗糖尿病提供了一种新方法。 方法：（1）取昆明种小鼠，分对照与实验组，适应性喂养后实验组小鼠按40mg/kg体重注射STZ，对照组注射相应体积柠檬酸缓冲液,连续注射5天，3天后测血糖值，取血糖值≥11.1mmol/L为成功模型。成模小鼠分为两组，一组灌胃富铬酵母悬液4周，另一组灌胃蒸馏水4周，测血糖值。（2）取昆明种小鼠，分对照与实验组，适应性喂养后实验组小鼠按200mg/kg体重注射STZ，对照组注射相应体积柠檬酸缓冲液，3d后测血糖，取血糖值≥11.1mmol/L为成功模型。成模小鼠分为两组，一组灌胃富铬酵母悬液4周，另一组灌胃蒸馏水4周，测血糖值。（3）取C57BL/6J断乳小鼠，随机分为正常饲料组和高脂饲料组，分别用相应饲料喂养3 周。高脂饲料组又分为高脂饲料对照组和高脂饲料实验组。第3 周末, 高脂饲料实验组腹腔内按100mg/kg体重一次性腹腔注射STZ；正常饲料组和高脂饲料对照组腹腔注射相应体积的无菌柠檬酸缓冲液。继续喂养4 周。小鼠以第7周末血糖为准，≥11.1mmol/L为成功模型。成模小鼠分为2组，1组每日灌胃富铬酵母悬液，另一组灌服相应体积的去离子水，4周后，测血糖值。 结果：对Ⅰ型糖尿病小鼠，富铬酵母治疗2周后，治疗组血糖明显低于对照组血糖（p<0.05），4周后显著低于(p<0.01)；对Ⅱ型糖尿病小鼠，富铬酵母治疗2周后，治疗组血糖明显低于对照组血糖（p<0.05），3周后显著低于(p<0.01)；对肥胖引起的Ⅱ型糖尿病小鼠，富铬酵母治疗2周后，治疗组血糖显 著低于对照组血糖(p<0.01)，且血清胰岛素浓度之间没有明显差异。 结论：富铬酵母具有明显的降血糖作用，且不刺激胰岛素分泌

Discrimination of Type 2 diabetic patients from healthy controls by using metabonomics method based on their serum fatty acid profiles

Metabonomics, the study of metabolites and their roles in various disease states, is a novel methodology arising from the post-genomics era. This methodology has been applied in many fields, including work in cardiovascular research and drug toxicology. In this study, metabonomics method was employed to the diagnosis of Type 2 diabetes mellitus (DM2) based on serum lipid metabolites. The results suggested that serum fatty acid profiles determined by capillary gas chromatography combined with pattern recognition analysis of the data might provide an effective approach to the discrimination of Type 2 diabetic patients from healthy controls. And the applications of pattern recognition methods have improved the sensitivity and specificity greatly. (C) 2004 Elsevier B.V. All rights reserved.

Screening and Structural Characterization of alpha-Glucosidase Inhibitors from Hawthorn Leaf Flavonoids Extract by Ultrafiltration LC-DAD-MSn and SORI-CID FTICR MS

In vitro a-glucosidase inhibition assays and ultrafiltration liquid chromatography with photodiode array detection coupled to electrospray ionization tandem mass spectrometry (ultrafiltration LC-DAD-ESI-MSn) were combined to screen a-glucosidase inhibitors from hawthorn leaf flavonoids extract (HLFE). As a result, four compounds were identified as alpha-glucosidase inhibitors in the HLFE, and their structures were confirmed to be quercetin-3-O-rha-(1-4)-glc-rha and C-glycosylflavones (vitexin-2 ''-O-glucoside, vitexin-2 ''-O-rhamnoside and vitexin) by high-resolution sustained off resonance irradiation collision-induced dissociation (SORI-CID) data obtained by Fourier transform ion cyclotron resonance mass spectrometry (FTICR MS).

Glucose-lowering activity of amino acid-N-phosphonic acid oxovanadium complexes and its interaction with DNA

Vanadium has well-documented lowering glucose properties both in vitro and in vivo. The design of new oxovanadium(IV) coordination compounds, intended for use as insulin-enhancing agents in the treatment of diabetes mellitus, can potentially benefit from a synergistic approach, in which the whole complex has more than an additive effect from its component parts. Biological testing with oxovanadium(IV) organic phosphonic acid, for insulin-enhancing potential included acute administration, by oral gavage in streptozotocin (STZ) diabetic rats. The complexes of oxovanadium(IV) amino acid-N-phosphonic acid exhibit higher lowering glucose activity in vivo. The interaction of the complexes of oxovanadium(IV) amino acid-N-phosphonic acid with DNA was investigated by agarose gel electrophoresis. The results indicated that these complexes have strong interaction with DNA.

海带根中α-glucosidase 和PTP-1B 靶向生物活性物质的分离纯化及其抗糖尿病作用研究

海带根是一种治疗糖尿病的民间中药，在沿海地区有很长的民间用药历史。食用海带根能够有效降低糖尿病患者的血糖，起到治疗作用。本文目的在于发现海带根中抗糖尿病的天然活性物质并分析它们在糖尿病治疗中的靶点；进一步开发一种低价且无毒副作用的化学类新药或中药新药。 α-glucosidase和 PTP-1B是II型糖尿病的两个重要靶点，海带根提取物能同时作用于这两个靶点。通过抑制这两种酶，降低血糖水平，85%乙醇粗提物对两种酶的IC50分别为1589ug/ml、IC50 1271ug/ml。乙酸乙酯相和石油醚相分别抑制α-glucosidase和 PTP-1B，IC50分别为380ug/ml和220ug/ml。因此以α-glucosidase和 PTP-1B的抑制活性为导向，用天然产物化学的方法对活性成分进行追踪分离，寻找单体活性物质进而鉴定其结构。由于乙酸乙酯相具有α-glucosidase抑制活性，用硅胶柱层析（石油醚：丙酮5：1、1：1），（二氯甲烷：甲醇60：1、20：1、5：1），凝胶柱层析Sephadex LH20（二氯甲烷：甲醇1：1），HPLC （80% 甲醇-水），对α-glucosidase抑制剂进行分离，得到组分IC50 为3.6ug/ml。用质谱仪和核磁共振确定结构。 生物活性测定结果表明α-glucosidase和 PTP-1B是两种不同的物质，分别位于乙酸乙酯相和石油醚相。光照实验和高温实验表明抑制α-glucosidase的活性成分对光照和温度敏感。光照48h或者50℃ 12h而且对α-glucosidase的抑制活性显著降低，TLC检测并用FeCl3显色初步表明抑制α-glucosidase的活性成分可能是多数酚类物质。动物实验显示在1450ug/kg剂量下，乙酸乙酯相能够显著降低糖尿病小鼠血糖，与阴性对照组差异极显著（P<0.01）。表明，海带根提取物在体内和体外均呈现出抗糖尿病活性，是一种潜在的抗糖尿病药物。

Total synthesis of bis-(2,3-dibromo-4,5-dihydroxyphenyl)-methane as potent PTP1B inhibitor

Protein tyrosine phosphatase 1B (PTP1B) plays an important role as a negative regulator and has been proved to be an effective target for the treatment of type 2 diabetes mellitus. Bis-(2,3-dibromo-4,5-dihydroxyphenyl)-methane 7 was first reported as a natural bromophenol with significant inhibition against PTP1B which was isolated from red algae Rhodomela confervoides. Intrigued by its astonishing activity (IC50 = 2.4 mu mol/L), compound 7 was synthesized with the overall yield of 24% and evaluated for its PTP1B inhibitory activity compared with natural compound. (C) 2008 Li Jun Han. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Inhibition of bromophenols against PTP1B and anti-hyperglycemic effect of Rhodomela confervoides extract in diabetic rats

Protein tyrosine phosphatase 1B (PTP1B) plays an important role as a negative regulator in insulin signaling pathways. PTP1B is an effective target for the treatment of type 2 diabetes mellitus. Four bromophenol derivatives from red algae Rhodomela confervoides, 2,2',3,3'-tetrabromo-4,4',5,5'-tetra-hydroxydiphenyl methane (1), 3-bormo-4,5-bis(2,3-dibromo-4,5-dihydroxybenzyl) pyrocatechol (2), bis(2,3-dibromo-4,5-dihydroxybenzyl) ether (3) and 2,2',3-tribromo-3',4,4',5-tetrahydroxy-6'-ethyloxy-methyldiphenylmethane (4) showed significant inhibitory activity against PTP1B (IC50 were 2.4, 1.7, 1.5 and 0.84 mu mol/L, respectively) as potential therapeutical agents for the treatment of type 2 diabetes mellitus. The anti-hyperglycemic effects of the ethanol extracts from R. confervoides on streptozotocin-induced diabetes (STZ-diabetes) in male Wistar rats fed with high fat diet were investigated. The STZ-diabetic rats treated with medium-dose and high-dose alga extracts showed remarkable reductions in fasting blood glucose (FBG) as compared with the STZ-diabetic control. The results indicate that the in vivo anti-hyperglycemic activity of the R. confervoides extracts can be partially attributed to the inhibitory actions against PTP1B of the bromophenol derivatives and that may be of clinical importance in improving the management of type 2 diabetes mellitus.

中老年人工作记忆与血管性疾病及危险因素的相关分析

Background With the increasing of an elderly population, the number of both vascular disease and cognitive function is increasing. Recent study shown that there is close relationship between vascular disease or vascular risk factors and cognitive function, but many questions are unclear. Objective The aim of this study was to explore the status of cognitive function of midlife and elderly subjects and to evaluate association between vascular risk factors and cognitive function. Methods We selected 330(female,128,male,202) retire navy veterans aged between 55 and 87 years old in Beijing. The average age was 67 years old. Data about vascular risk factors, including: history of cerebral infarction, cardiovascular disease, hypertension, diabetes mellitus, carotid lesions, cigarette smoking, drinking, obesity, and so on. Cognitive functions were assessed by neuropsychological tests: the four tasks of working memory. Results The univariate analysis indicated that there were significant differences of working memory scores in aging, gender, education level, the history of stroke, hypertension with midlife and elderly subjects. Moderate levels of alcohol intake maybe better for cognition. Conclusion vascular disease and some vascular risk factors may lead to cognitive function decline, thus, intervention methods should be carried out in middle and elderly people with vascular risk fact should be under special supervision, with at least annual neuropsychological evaluation. Key words: cognitive function; vascular risk factor， working memory.