血镁水平与糖尿病肾病

为探讨糖尿病肾病 (DN)患者血镁水平 ,以 1 5 9例 2型糖尿病 (T2DM)患者为研究对象 ,用xylidylblue比色法测定了其正常白蛋白尿期、微量白蛋白尿期 (早期DN)、临床DN及晚期DN血镁水平 ,与 2 0例正常对照组相比较 ;同时将 1 1 0例DN患者分为肾功能不全代偿组、失代偿组和肾功能衰竭组 ,将其血镁水平与 1 0 2例慢性肾小球肾炎 (CGN)患者相比较 ,组间比较采用t检验 ;对 79例T2DM正常白蛋白尿期及微量白蛋白尿期患者尿白蛋白排泄率 (UAER)与血镁水平进行了等级相关分析。结果表明 ,T2DM正常白蛋白尿期、早期DN及临床DN血镁水平降低 ,晚期DN血镁水平升高 ,各组与正常对照组相比存在显著性差异 (P <0 0 5或P <0 0 1 ) ;T2DMDN及CGN肾功能不全代偿期、失代偿期及肾功能衰竭期血镁水平渐升高 ,且肾功能处于同一期的DN和CGN相比 ,前者血镁水平均较后者显著降低 (P <0 0 1或P <0 0 5 ) ;T2DM正常白蛋白尿期和微量白蛋白尿期血镁水平与UAER呈负相关 (r=0 5 47,P <0 0 1 )。提示T2DM患者肾功能正常时存在低镁倾向 ,但晚期DN血镁水平升高 ;随着肾功能不全进展 ,DN和CGN患者血镁水平升高 ,但前者血镁水平仍较后者低 ,镁代谢紊乱与DM及其并发症的相互关系有待进一步研究 ;血镁水平测定可否作为DN的早期诊断指标亦     

探讨尿蛋白和尿微量白蛋白在糖尿病肾病检验中的应用价值

目的对尿蛋白和尿微量白蛋白在糖尿病肾病检验中的应用价值进行评价分析,为今后的临床诊断工作提供有价值的参考信息。方法选择2013年5月—2015年5月间江西省上饶市广丰县中医院收治的符合临床诊断标准的糖尿病肾病患者56例以及同期健康体检者60例作为研究对象,对这两组受试者展开尿蛋白和尿微量白蛋白水平检测,并对检测结果进行对比分析。结果糖尿病肾病组患者尿蛋白、尿微量白蛋白阳性率较健康对照组发生显著升高(P0.05)。结论对患者展开尿蛋白和尿微量白蛋白水平检测,为临床糖尿病肾病的早期诊断提供可靠的参考依据,临床价值显著,值得关注并推广。     

微量元素锌、血清胱抑素 C在糖尿病肾病早期诊断的意义

目的：探讨早期糖尿病肾病患者（ DN）的微量元素锌、血清胱抑素C（ CysC）、血清肌酐（ SCr）和血清尿素（ BUN）水平的变化程度及意义。方法将糖尿病肾病患者分为3组：正常蛋白尿组,微量蛋白尿组,临床蛋白尿组。所有患者和对照组均空腹抽取静脉血检测微量元素锌、血清胱抑素C、血清尿素、血清肌酐。结果正常蛋白尿组、微量蛋白尿组、临床蛋白尿组微量元素锌均明显低于正常对照组,差异有统计学意义（ P＜0.05）。正常蛋白尿组、微量蛋白尿组、临床蛋白尿组血清胱抑素C水平均明显高于正常对照组,差异有统计学意义（ P＜0.05）。微量蛋白尿组、临床蛋白尿组血清肌酐检测值和正常对照组有明显差异,差异有统计学意义（ P＜0.05）。结论检测微量元素锌、血清胱抑素C对糖尿病肾病的早期肾损害有较高的临床价值,可作为早期糖尿病肾病诊断的参考指标。     

空腹血糖以及糖化血红蛋白对糖尿病前期患者的诊断效果评价

目的探讨空腹血糖(FPG)与糖化血红蛋白(Hb A1c)对临床中糖尿病前期的诊断价值。方法将黑龙江省佳木斯大学第一附属医院接收治疗的120例疑似糖尿病患者作为研究分析对象,将所选患者按诊断方式分为对照组和研究组(n=60),对照组采用糖化血红蛋白(Hb A1c)进行诊断,研究组采用空腹血糖(FPG)进行诊断,比较两组诊断正确率及两年内发展为糖尿病的发病率。结果研究组42例(70.0%)被确诊为糖尿病前期患者,18例(42.86%)患者发展为糖尿病;对照组中28例确诊为糖尿病前期患者,5例(17.85%)患者发展为糖尿病,确诊后转变率观察组高于对照组,差异有统计学意义(P0.05)。结论诊断糖尿病前期采用空腹血糖(FPG)与糖化血红蛋白(Hb A1c),前者临床价值更显著,建议将两种诊断方式联用,临床价值更大。     

黄芩-栀子配伍治疗糖尿病肾病大鼠的尿液代谢组学研究

采用基于超高效液相色谱与串联四极杆飞行时间质谱仪(Ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry,UHPLC-Q-TOF MS)联用的代谢组学方法,通过分析大鼠尿液内源性代谢物的变化,研究黄芩-栀子药对治疗糖尿病肾病的作用机制。利用高脂高糖饲料喂养并腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型。给药12周后,采用UHPLC-Q-TOF MS对正常对照组、模型组、黄芩-栀子药对治疗组的尿液样品进行代谢轮廓分析。利用正交偏最小二乘判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)对实验数据进行分析,挑选出对各组分分离贡献较大(VIP>1,p<0.05)的化合物在Human Metabolome Database(HMDB)等数据库进行质谱信息匹配,共鉴定出糖尿病肾病潜在生物标记物31个,给药后回调生物标记物16个。研究结果表明,黄芩-栀子药对通过影响胆汁酸的合成与代谢、尿毒素代谢、能量代谢、色氨酸代谢、苯丙氨酸代谢、酪氨酸代谢等通路对糖尿病肾病起到治疗作用,其中胆汁酸的合成与代谢、尿毒素代谢和能量代谢可能是黄芩-栀子药对发挥治疗作用的主要途径。与单味药相比,配伍后的药对具有了新的药理作用。     

五味子治疗大鼠糖尿病肾病作用机制的血清代谢组学研究

采用基于超高效液相色谱与串联四级杆飞行时间质谱仪(Ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry,UPLC/Q-TOF-MS)联用技术的代谢组学方法,通过分析大鼠血清内源性代谢物的变化,研究五味子治疗糖尿病肾病的作用机制。利用高脂高糖饲料喂养并腹腔注射链脲佐菌素(STZ)建立糖尿病大鼠模型。给药12周后,采用试剂盒方法测定尿蛋白、尿肌酐的含量,结果表明五味子水提取物可以显著降低模型动物的尿蛋白含量(p<0.05),对糖尿病大鼠肾病并发症具有一定的改善作用。采用UPLC/Q-TOF-MS方法分析了五味子对糖尿病肾病大鼠的血清代谢轮廓,分析了健康组、模型组和五味子给药组的大鼠血清,采用偏最小二乘法-判别分析(Partial least squares discriminant analysis,PLS-DA)进行数据分析。PLS-DA得分图显示健康组、模型组和五味子组的代谢轮廓有显著差别,根据正交偏最小二乘法-判别分析(Orthogonal partial least squares discriminant analysis,OPLS-DA)载荷图筛选,将对各组分离贡献大的化合物的串联质谱分析数据,经Human Metabolome Database(HMDB)等数据库检索,进行质谱信息匹配,鉴定出黄尿酸、油酰胺、棕榈酰胺、尿酸、5-羟基己酸、硫酸对甲酚、对甲酚葡萄糖苷酸7种内源性代谢物为生物标记物。研究结果表明五味子通过影响色氨酸代谢、嘌呤代谢、肠内菌代谢、脂肪酸代谢等通路对糖尿病肾病发挥治疗作用,其中嘌呤代谢、肠内菌代谢通路可能是五味子发挥治疗作用的重要途径。     

DN非蛋白氮色谱分离检测新技术开发成功

不久前，西安交通大学生物医学信息工程教育部重点实验室与第四军医大学药学系、西安理工大学应用化学系联合开发出一种“糖尿病肾病非蛋白氮代谢物反相高效液相色谱分离检测”新技术。该技术的开发成功，为临床糖尿病肾病患者的早期诊断提供了理想的检测手段。     

MRA技术在破裂颅内动脉瘤诊断及术后复发预测效能中的应用价值

选取颅内动脉瘤(IA)破裂患者45例为破裂组,同期IA未破裂患者81例为未破裂组,均接受磁共振血管成像(MRA)技术检查及血管内栓塞术治疗,并根据术后3个月有无复发情况分为复发者和无复发者进行对比研究。结果发现,动脉瘤最长径与瘤颈宽度比值(AR)、标准化表面最大剪切力(NWSSM)、剪切力震荡指数(OSI)联合诊断IA破裂的AUC值最大;动脉瘤大小、即刻栓塞效果、AR、NWSSM、OSI是IA术后复发的重要影响因素;AR、NWSSM、OSI联合预测IA术后复发的敏感度、特异度分别为76.47%、90.83%。提示采用MRA技术监测IA形态学与血流动力学,可为临床诊断IA破裂、预测术后复发提供科学循证指导。     

一种具有光致变色特性的膜蛋白/聚合物复合膜

通过基因工程技术将古紫质4(archaerhodopsin-4,AR4)膜蛋白的基因转入本身不表达该蛋白质、也不表达玉红素的嗜盐菌株L33中,去除了天然菌株xz515所获得的AR4膜蛋白脂环境中的玉红素,优化了AR4的光致变色性能.将重组古紫质4包埋于聚乙烯醇中制成复合膜,并通过提高体系的pH值,延长了M态的寿命,进一步实现了简单图案记录,验证了该活性蛋白质与聚合物的复合材料具有光信息存储功能.     

高强度双网络水凝胶的增韧机理

双网络水凝胶(DN凝胶)是由具有很强的结构非对称性的两种聚合物形成的特殊的聚合物互穿网络。相对于单一聚合物网络(SN凝胶)而言,DN凝胶的机械强度和韧性都有惊人的提高,其拉伸断裂应力和断裂应变分别能达到1~10 MPa和1000%~2000%、韧性(撕裂能)可以达到102~103J·m-2。DN凝胶性能的提高,主要是由强对称结构的第一重网络(刚而脆)与第二重网络(软而韧)相互缠结和互穿的结果。对DN凝胶断裂过程和增韧本质的理解是设计下一代具有理想机械性能的DN凝胶的关键所在。一些DN凝胶表现出大的滞后、屈服、细颈和软化现象,而这些现象用经典的Lake-Thomas断裂理论是没有办法解释的。根据DN凝胶的滞后和细颈等实验现象,Brown和Tanaka等提出了"破坏区"理论来解释DN凝胶超乎寻常的高韧性。最近,龚剑萍等提出的"牺牲键"理论也已经很好地应用于设计和制备具有新型微纳米结构的高韧性DN凝胶。本文着重阐释了DN凝胶的增韧机理,总结了这一领域的最新研究成果,并讨论各种因素对凝胶韧性的影响,最后对DN凝胶增韧机理存在的问题和研究方向进行了展望。     

Angiotensin-I converting enzyme gene polymorphism in Turkish type 2 diabetic patients

Non-insulin dependent diabetes mellitus is often associated with some complications such as nephropathy, retinopathy and neuropathy. Genes of the renin angiotensin system are potential candidate genes for diabetic complications. We investigated the relationship between angiotensin converting enzyme (ACE) gene polymorphism in type 2 diabetic patients with and without diabetic nephropathy. Seventy five patients (25 type 2 diabetic patients with nephropathy, 50 type 2 diabetic patients without nephropathy) and 37 healthy controls were studied. Gene polymorphism of ACE was determined by PCR (polymerase chain reaction) amplification using allele-spesific primers. The frequencies of ACE DD, ID and II genoypes among the patients with type 2 diabetic patients were found 48%, 42%, 10% whereas in control subjects, 27%, 60%, 13% respectively. Type 2 diabetic patients carrying DD genotype without nephropathy increased 1.77 fold than control subjects (P < 0.05). There is no significant correlation between diabetic nephropathy and ACE gene polymorphism. But we found that ACE DD genotype increased significantly in type 2 diabetic patients compared to control subjects (P <.05).     

Plasma esterified and non-esterified fatty acids metabolic profiling using gas chromatography-mass spectrometry and its application in the study of diabetic mellitus and diabetic nephropathy

Using gas chromatography–mass spectrometry (GC–MS), a new metabolic profiling method was established to assess the levels of non-esterified fatty acids (NEFAs) and esterified fatty acids (EFAs) in plasma. The extraction method was simple and robust without removing protein process. With this method 25 fatty acids (FAs), both EFAs and NEFAs, can be recognized simultaneously with only 10 μL plasma. 15 of the 25 can be precisely quantified. The method was validated and then applied into clinical metabonomics research. Five clinical groups including 150 cases were involved. The relationship between FA levels and diabetic mellitus (DM) as well as diabetic nephropathy (DN) pathology was speculated. Furthermore, the possible pathological causes and effects were discussed in detail. Potential biomarkers (p value <0.01) were screened with Student's t-test. With the application of partial least squares-discriminant analysis (PLS-DA), different stages were distinguished. The result may be useful for the pathology study of metabolic syndromes, and may also be helpful for monitoring the progression of DM and DN.     

Anti-diabetic Properties of Calcium Channel Blockers: Inhibition Effects on Aldose Reductase Enzyme Activity

Applied Biochemistry and Biotechnology - Aldose reductase (AR) belongs to NADPH-dependent oxidoreductases and converts glucose to sorbitol in the polyol pathway. AR inhibition is essential to...     

Metabolic profiling reveals new serum biomarkers for differentiating diabetic nephropathy

Capillary electrophoresis coupled with time-of-flight mass spectrometry was used to explore new serum biomarkers with high sensitivity and specificity for diabetic nephropathy (DN) diagnosis, through comprehensive analysis of serum metabolites with 78 diabetic patients. Multivariate analyses were used for identification of marker candidates and development of discriminative models. Of the 289 profiled metabolites, orthogonal partial least-squares discriminant analysis identified 19 metabolites that could distinguish between DN with macroalbuminuria and diabetic patients without albuminuria. These identified metabolites included creatinine, aspartic acid, γ-butyrobetaine, citrulline, symmetric dimethylarginine (SDMA), kynurenine, azelaic acid, and galactaric acid. Significant correlations between all these metabolites and urinary albumin-to-creatinine ratios (p?<?0.009, Spearman’s rank test) were observed. When five metabolites (including γ-butyrobetaine, SDMA, azelaic acid and two unknowns) were selected from 19 metabolites and applied for multiple logistic regression model, AUC value for diagnosing DN was 0.927 using the whole dataset, and 0.880 in a cross-validation test. In addition, when four known metabolites (aspartic acid, SDMA, azelaic acid and galactaric acid) were applied, the resulting AUC was still high at 0.844 with the whole dataset and 0.792 with cross-validation. Combination of serum metabolomics with multivariate analyses enabled accurate discrimination of DN patients. The results suggest that capillary electrophoresis-mass spectrometry based metabolome analysis could be used for DN diagnosis.     

Development of Novel Indole-Based Bifunctional Aldose Reductase Inhibitors/Antioxidants as Promising Drugs for the Treatment of Diabetic Complications

Aldose reductase (AR, ALR2), the first enzyme of the polyol pathway, is implicated in the pathophysiology of diabetic complications. Aldose reductase inhibitors (ARIs) thus present a promising therapeutic approach to treat a wide array of diabetic complications. Moreover, a therapeutic potential of ARIs in the treatment of chronic inflammation-related pathologies and several genetic metabolic disorders has been recently indicated. Substituted indoles are an interesting group of compounds with a plethora of biological activities. This article reviews a series of indole-based bifunctional aldose reductase inhibitors/antioxidants (ARIs/AOs) developed during recent years. Experimental results obtained in in vitro, ex vivo, and in vivo models of diabetic complications are presented. Structure–activity relationships with respect to carboxymethyl pharmacophore regioisomerization and core scaffold modification are discussed along with the criteria of ‘drug-likeness”. Novel promising structures of putative multifunctional ARIs/AOs are designed.     

Protective Effects of Swertiamarin against Methylglyoxal-Induced Epithelial-Mesenchymal Transition by Improving Oxidative Stress in Rat Kidney Epithelial (NRK-52E) Cells

Increased blood glucose in diabetic individuals results in the formation of advanced glycation end products (AGEs), causing various adverse effects on kidney cells, thereby leading to diabetic nephropathy (DN). In this study, the antiglycative potential of Swertiamarin (SM) isolated from the methanolic extract of E. littorale was explored. The effect of SM on protein glycation was studied by incubating bovine serum albumin with fructose at 60 °C in the presence and absence of different concentrations of swertiamarin for 24 h. For comparative analysis, metformin was also used at similar concentrations as SM. Further, to understand the role of SM in preventing DN, in vitro studies using NRK-52E cells were done by treating cells with methylglyoxal (MG) in the presence and absence of SM. SM showed better antiglycative potential as compared to metformin. In addition, SM could prevent the MG mediated pathogenesis in DN by reducing levels of argpyrimidine, oxidative stress and epithelial mesenchymal transition in kidney cells. SM also downregulated the expression of interleukin-6, tumor necrosis factor-α and interleukin-1β. This study, for the first time, reports the antiglycative potential of SM and also provides novel insights into the molecular mechanisms by which SM prevents toxicity of MG on rat kidney cells.     

HPLC-electrospray tandem mass spectrometry for simultaneous quantitation of eight plasma aminothiols: Application to studies of diabetic nephropathy

It was reported that Hcy was related to the development of kidney disease, but it remains unknown whether Hcy is an independent biomarker for diabetic nephropathy. Analytical method for simultaneous determination of aminothiols among the Hcy metabolic cycle is desirable to discover other potential biomarkers. A high-performance liquid chromatography-electrospray tandem mass spectrometric (HPLC-ESI-MS/MS) method was established for simultaneous quantitation of Cysteine (Cys), total homocysteine (tHcy), S-adenosylmethionine (SAM), S-adenosylhomocysteine (SAH), cystathionine (Cysta), methionine (Met), glutathione (GSH) and cysteinylglycine (Cys-gly) in plasma with N-(2-mercaptopropionyl)-glycine (MPG) as internal standard. The method had simple pretreatment without derivatization and the chromatograms show better separation of the eight aminothiols and the analytic time was 20 min. The results demonstrated that it provided an excellent linearity for all analytes over their respective concentration ranges and illustrated excellent precision and plasma recovery as well. Then, the method was applied in the case-control study of patients with diabetes mellitus (DM) and diabetic nephropathy (DN). In conclusion, it is an effective method to quantitate the concentrations of aminothiols in the human plasma. SAH and SAM were suggested as better potential biomarkers of DM and DN.     

基于UPLC-oaTOF-MS的糖尿病及糖尿病肾病的代谢组学研究

以糖尿病患者、糖尿病肾病患者和正常人的血清为研究对象, 采用超高效液相色谱-飞行时间质谱建立其代谢指纹图谱, 并结合主成分分析进行模式识别, 实现患者和正常人的区分, 并试图发现潜在的标志物.