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.     

毛细管电泳间接电致化学发光灵敏检测去甲肾上腺素及其在尿样分析中的应用

基于3种肾上腺素对三联吡啶钌-三丙胺发光体系具有强烈抑制的特性, 建立了一种毛细管电泳分离-间接电致化学发光(CE-IECL)灵敏检测NE、脱氧肾上腺素和异丙肾上腺素的新方法, 并将该方法成功地应用于人尿样分析. 结果表明, 糖尿病肾病患者尿样中的NE含量显著比健康人的高.     

Combined Discrete-event Simulation and Ant Colony Optimisation Approach for Selecting Optimal Screening Policies for Diabetic Retinopathy

In this paper we present the first application to a healthcare problem of discrete-event simulation (DES) embedded in an ant colony optimisation (ACO) model. We are concerned with choosing optimal screening policies for retinopathy, a sight-threatening complication of diabetes. The early signs of retinopathy can be detected by screening before the patient is aware of symptoms, and blindness prevented by laser treatment. In this paper we describe the methodology used to combine the purpose-written DES model with the ACO algorithm. We simulate the effects of different screening strategies on a population of diabetic patients, and compare them in terms of two objective functions: Min C/E, cost-effectiveness (minimum incremental cost per year of sight saved, compared with a no-screening baseline) and Max E, maximum effectiveness (years of sight saved). We describe how ACO is used to optimise these two objectives, and discuss the issues involved in optimising stochastic variables. We present results for a range of different assumptions and scenarios about the format of screening programmes, using realistic data, and make policy recommendations on the basis of our findings.     

The reno-protective effect of a phosphoinositide 3-kinase inhibitor wortmannin on streptozotocin-induced proteinuric renal disease rats

Diabetic nephropathy (DN) is a progressive kidney disease that is caused by injury to kidney glomeruli. Podocytes are glomerular epithelial cells and play critical roles in the glomerular filtration barrier. Recent studies have shown the importance of regulating the podocyte actin cytoskeleton in early DN. The phosphoinositide 3-kinase (PI3K) inhibitor, wortmannin, simultaneously regulates Rac1 and Cdc42, which destabilize the podocyte actin cytoskeleton during early DN. In this study, in order to evaluate the reno-protective effects of wortmannin in early DN by regulating Rac1 and Cdc42, streptozotocin (STZ)-induced proteinuric renal disease (SPRD) rats were treated with wortmannin. The albuminuria value of the SPRD group was 3.55 ± 0.56 mg/day, whereas wortmannin group was 1.77 ± 0.48 mg/day. Also, the albumin to creatinine ratio (ACR) value of the SPRD group was 53.08 ± 10.82 mg/g, whereas wortmannin group was 20.27 ± 6.41 mg/g. Changes in the expression level of nephrin, podocin and Rac1/Cdc42, which is related to actin cytoskeleton in podocytes, by wortmannin administration were confirmed by Western blotting. The expression levels of nephrin (79.66 ± 0.02), podocin (87.81 ± 0.03) and Rac1/Cdc42 (86.12 ± 0.02) in the wortmannin group were higher than the expression levels of nephrin (55.32 ± 0.03), podocin (53.40 ± 0.06) and Rac1/Cdc42 (54.05 ± 0.04) in the SPRD group. In addition, expression and localization of nephrin, podocin and desmin were confirmed by immunofluorescence. In summary, we found for the first time that wortmannin has a reno-protective effect on SPRD rats during the early DN. The beneficial effects of wortmannin in SPRD rats indicate that this compound could be used to delay the progression of the disease during the early DN stage.     

Creation of Chitosan-Based Nanocapsule-in-Nanofiber Structures for Hydrophobic/Hydrophilic Drug Co-Delivery and Their Dressing Applications in Diabetic Wounds

Nanofiber meshes (NFMs) loaded with therapeutic agents are very often employed to treat hard-to-heal wounds such as diabetic wounds. However, most of the NFMs have limited capability to load multiple or hydrophilicity distinctive-therapeutic agents. The therapy strategy is therefore significantly hampered. To tackle the innate drawback associated with the drug loading versatility, a chitosan-based nanocapsule-in-nanofiber (NC-in-NF) structural NFM system is developed for simultaneous loading of hydrophobic and hydrophilic drugs. Oleic acid-modified chitosan is first converted into NCs by the developed mini-emulsion interfacial cross-linking procedure, followed by loading a hydrophobic anti-inflammatory agent Curcumin (Cur) into the NCs. Sequentially, the Cur-loaded NCs are successfully introduced into reductant-responsive maleoyl functional chitosan/polyvinyl alcohol NFMs containing a hydrophilic antibiotic Tetracycline hydrochloride. Having a co-loading capability for hydrophilicity distinctive agents, biocompatibility, and a controlled release property, the resulting NFMs have demonstrated the efficacy on promoting wound healing either in normal or diabetic rats.     

Pectin-Lyase-Modified Ginseng Extract and Ginsenoside Rd Inhibits High Glucose-Induced ROS Production in Mesangial Cells and Prevents Renal Dysfunction in db/db Mice

Diabetes increases the incidence rate of chronic renal disease. Pectin-lyase-modified ginseng (GS-E3D), with enhanced ginsenoside Rd content, has been newly developed. In this study, renal protective roles of GS-E3D in type-2 diabetic db/db mice were investigated. The generation of reactive oxygen species (ROS) induced by high glucose (25 mM) was reduced by ES-E3D (75%) and ginsenoside Rd (60%). Diabetic db/db mice received 100 or 250 mg/kg/day of GS-E3D daily via oral gavage for 6 weeks. Albuminuria and urinary 8-hydroxy-2′-deoxyguanosine (8-OhdG, an oxidative stress marker) levels were increased in db/db mice and the levels recovered after GS-E3D treatment. In renal tissues, TUNEL-positive cells were decreased after GS-E3D treatment, and the increased apoptosis-related protein expressions were restored after GS-E3D treatment. Therefore, GS-E3D has a potent protective role in diabetes-induced renal dysfunction through antioxidative and antiapoptotic activities. These results may help patients to select a dietary supplement for diabetes when experiencing renal dysfunction.     

双嘧达莫联合厄贝沙坦对IgA肾病的疗效及其安全性分析

目的探究双嘧达莫联合厄贝沙坦对Ig A肾病的疗效及其安全性。方法采取回顾性分析方法对遵义医学院第五附属(珠海)医院2013年7月—2016年7月Ig A肾病患者88例资料进行收集分析,随机分为单药组和联合组。单药组采用单纯厄贝沙坦治疗;联合组采用双嘧达莫联合厄贝沙坦治疗。比较两组患者Ig A肾病治疗效果;治疗前后24 h尿蛋白、血肌酐;不良反应发生率。结果联合组患者Ig A肾病治疗效果比单药组高,P0.05;治疗前两组患者24 h尿蛋白、血肌酐差异不显著,P0.05;联合组治疗后24 h尿蛋白、血肌酐比单药组好,P0.05;联合组不良反应发生率和单药组差异不显著,P0.05。结论双嘧达莫联合厄贝沙坦对Ig A肾病的疗效及其安全性良好,可改善肾功能,无明显副作用,值得推广。     

毛细管区带电泳检测糖尿病肾病变患者血液内红细胞中的谷胱甘肽

采用高效毛细管区带电泳法对糖尿病肾病变患者血液内红细胞中参与氧化应激的氧化型谷胱甘肽(GSSG)和还原型谷胱甘肽(GSH)进行了测定。对影响分离的条件(缓冲液pH、缓冲液浓度、分离电压和毛细管温度)进行了优化,使用非涂层的毛细管(21 cm×75 μm i.d.)和20 mmol/L pH 6.86的磷酸缓冲液,在25 kV,30 ℃和200 nm条件下,可在3 min内同时对血红细胞中GSSG和GSH定量分析。方法具有良好的重现性(迁移时间和峰面积的相对标准偏差均小于2.0%)、较高的灵敏度(GSH:     

A UPLC–MS/MS method for comparative pharmacokinetics study of morusin and morin in normal and diabetic rats

The aim of this study was to establish and validate a rapid, selective and reliable ultra‐performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS) for simultaneous quantitations of morin and morusin, and to investigate their pharmacokinetics difference between normal and diabetic rats after oral administration. Plasma samples were pretreated via protein precipitation with acetonitrile. Genkwanin was used as internal standard (IS). Analytes and IS were separated on a Thermo Hypersil Gold C₁₈ column (50 × 4.6 mm, 3 μm) using gradient elution. The mobile phase consisted of acetonitrile and 0.1% formic acid in water at a flow rate of 0.5 mL/min. Mass spectrometry detection was carried out by means of negative electrospray ionization source and multipe‐reaction monitoring mode. The transitions of m/z 300.9 → 151.2 for morin, m/z 419.2 → 297.1 for morusin and m/z 283.1 → 268.2 for IS were chosen for quantification. Calibration curves were linear in the range of 1.01–504.2 ng/mL (r² ≥ 0.99) for morin and 1.02–522.3 ng/mL (r² ≥ 0.99) for morusin. The lower limit of quantification was 1.02 ng/mL for morin and 1.05 ng/mL for morusin. The extraction recovery was >85.1% for each analyte. No obvious matrix effect was observed under the present UPLC–MS/MS conditions during all of the bioanalysis. The stability study demonstrated that morin and morusin remained stable during the whole analytical procedure. The method was successfully applied to support the pharmacokinetic comparisons of morin and morusin between normal and diabetic rats.     

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.