微量元素与2型糖尿病的相关性研究

2型糖尿病是一种常见的内分泌及代谢紊乱疾病,人体必需微量元素直接影响细胞代谢和胰岛素的水平。综述了微量元素铬、锌、镁、硒、铁、铜与2型糖尿病（T2DM）相关性研究的进展,为临床糖尿病的预防和治疗提出建议。     

2型糖尿病与HER2过表达型乳腺癌预后的相关性研究

目的分析2型糖尿病与HER2过表达型乳腺癌预后的相关性。方法比较不合并2型糖尿的HER2过表达型乳腺癌患者与合并2型糖尿病的HER2过表达型乳腺癌患者在巩固期治疗后短期内的转移、复发与死亡率的差异。结果在HER2过表达型乳腺癌患者中,合并2型糖尿病的患者复发转移率与死亡率均高于不合并2型糖尿病的患者,差异有显著性(P0.05)。结论 2型糖尿病对HER2过表达型乳腺癌的预后不良有促进作用。     

基于磁性颗粒微阵列与双色荧光杂交的单核苷酸多态性分型方法研究

基于磁性颗粒微阵列与双色荧光杂交,建立了单核苷酸多态性(Single nucleoitide polymorphism,SNP)分型方法。将利用不对称扩增得到的含有待检测位点生物素标记的单链PCR产物固定在链亲和素修饰的金磁纳米颗粒(Gold magnetic nanoparticles,GMNPs)表面;将ssDNA-GMNPs混合物点样在底部固定有磁铁的载玻片上构建磁性颗粒微阵列,然后在基因框中与双色荧光探针杂交;杂交完全后,充分洗涤,通过扫描获得分型结果。通过优化不对称PCR的扩增条件,直接扩增出产量较高的单链DNA作为靶序列用于分型。利用本方法对24个样本MTHFR基因的C677T位点多态性进行了检测。实验证明,本方法步骤简单,易实现自动化操作、非常适用于分子诊断与法医鉴定。     

2型糖尿病患者血清微量元素分析

为探讨2型糖尿病与患者血清微量元素的关系,对105例2型糖尿病患者血清Fe、Zn、Cu、Cr、Mg、Ca进行了观察,并与正常对照组做比较,各指标间做相关分析,结果表明,2型糖尿病患者血清Zn、Cr含量明显低于对照组,Cu的含量明显高于对照组,Fe、Mg、Ca的含量两组无差异。提示糖尿病患者体内微量元素的含量与糖尿病具有密切的关系。     

基于多肽微阵列芯片的荧光和共振光散射法筛选凝血酶抑制剂

研制了一种基于多肽微阵列芯片的荧光和共振光散射双通路检测方法,对血液样品中的凝血酶抑制剂进行检测。以生物素化的多肽微阵列芯片为反应平台,加入凝血酶水解多肽上的特异位点使其C末端的生物素解离,通过亲和素和生物素之间的特异性结合,用荧光探针和30 nm金纳米粒子探针标记此反应过程。凝血酶抑制剂阻止凝血酶对底物多肽的水解反应,通过荧光和共振光散射信号强度的变化检测抑制剂的抑制能力。酶溶液和加标人血清中测定的半抑制浓度(IC50)值:阿加曲班<抗凝血酶Ⅲ<4-(2-氨乙基)苯磺酰氟盐酸盐,IC50差的绝对值随抑制剂特异性的减弱而增大。当抑制剂浓度为7.5μmol/L时,血浆中5种化合物的抑制能力:阿加曲班>抗凝血酶Ⅲ>胰蛋白酶抑制剂>N-(反式-环氧丁二酰基)-L-亮氨酸-4-胍基丁基酰胺>4-(2-氨乙基)苯磺酰氟盐酸盐。在血浆中考察了阿加曲班和抗凝血酶Ⅲ的可逆性。对比荧光法和共振光散射法在血液样品中的检测结果,用30 nm金纳米粒子标记的共振光散射法更适用于复杂血液样品中抑制剂的检测     

富含微量元素中药煎剂对2型糖尿病疗效观察

为了观察富含微量元素中药煎剂对2型糖尿病（T2DM）的疗效,将门诊64名血糖控制不满意的T2DM患者分为增加富含微量元素中药煎剂组（观察组）和不增加组（对照组）,服药3个月后比较。结果表明,观察组的空腹血糖（FPG）、空腹血清胰岛素（Fins）和糖化血红蛋白（HbA1c）3月后明显下降（P〈0．01）;3月后观察组的FPG、Fins和HbA1c较对照组明显下降（P〈0．01）。可见增加富含微量元素中药煎剂对血糖难以控制的T2DM的FPG、Fins和HbA1c能够起到有效控制作用。     

两种胰岛素方案治疗门诊初诊2型糖尿病患者的疗效及低血糖发生率比较

目的研究两种胰岛素方案治疗门诊初诊2型糖尿病患者的疗效及低血糖发生率。方法选取孝感市中心医院2013年3月—2014年8月84例门诊初诊2型糖尿病患者,抽签随机分为两组,其中42例采用餐前皮下注射门冬胰岛素治疗记为对照组,剩下42例采用餐前注射门冬胰岛素联合晚间注射甘精胰岛素强化治疗记为观察组。结果两组治疗后血糖相关指标比较无明显差异,不具有统计学意义(P0.05);观察组低血糖率11.90%较对照组30.95%显著较低,具有统计学意义(P0.05)。结论两种胰岛素方案治疗初诊2型糖尿病均较为有效,但早期采用强化治疗方案能有效稳定平衡血糖代谢、预防低血糖发生,可作为临床治疗初诊2型糖尿病的首选参考方法。     

化学生物学中光敏分子微阵列的表面构建与应用

分子微阵列是有机合成（特别是组合化学合成）方法应用于生物和医学研究而发展起来的高科技集成技术,通过把微电子、微加工技术和有机化学合成反应相结合,在固体基质（如硅片、玻片、瓷片等）表面构建微型的生物有机化学分子系统,以实现对细胞、蛋白质、核酸及其他生物组分进行快速、敏感、高效地处理．近年来,随着表面化学构建策略研究的不断深入和迅猛发展,分子微阵列技术的应用领域不断拓展,已从最初用于核酸分子的杂交测序延伸到基因组功能研究的各个方面．本文着重综述了光敏分子微阵列的表面化学构建策略研究及其在化学生物学分析中应用的最新进展,并展望了其发展的未来趋势．内容主要包括：小分子与多肽分子微阵列、蛋白质分子微阵列、核酸分子微阵列和糖分子微阵列等．     

基于氧化镍沉积硅胶固相萃取与液相色谱-质谱联用技术的2型糖尿病血清中咪唑丙酸的检测

咪唑丙酸可以通过哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)路径引起人的胰岛素抵抗,从而导致2型糖尿病(T2D),因此,咪唑丙酸的准确定量可辅助2型糖尿病的诊断。本文利用NiO与咪唑基团之间的配位作用,采用氧化镍沉积硅胶(NiO@SiO₂)萃取材料对咪唑丙酸进行选择性富集和萃取。首先对NiO@SiO₂固相萃取(SPE)条件进行优化:吸附剂用量为200 mg,上样液为20 mmol/L的磷酸盐缓冲溶液(pH=3. 0),解吸液为1. 0 mL含1%(质量分数) NH₃·H₂O的水溶液;然后,对萃取液进行高效液相色谱-质谱(HPLC-MS)分离分析,建立了血清中咪唑丙酸的检测方法。结果表明,咪唑丙酸浓度在0. 05～10 ng/mL范围内对质谱响应值具有良好的线性关系(R²≥0. 996),检出限和定量限分别为0. 02和0. 05 ng/mL,加标回收率为84. 0%～119%,相对标准偏差RSD<17. 2%。将该方法用于检测2型糖尿病患者与正常人血清样品中的咪唑丙酸,...     

高灵敏纳米金比色芯片检测乙型肝炎病毒DNA及其变异

构建了新型纳米金比色芯片,利用Taq DNA连接酶的连接特异性,将其与乙型肝炎病毒DNA( HBV-DNA)靶序列完全互补杂交的捕获探针(固定在芯片上)和纳米金修饰的探针连接成一条链,从而将纳米金颗粒固定到芯片点阵上,再通过银染反应放大,形成裸眼可见的显色信息.通过点阵的位置及灰度,即可判断HBV-DNA靶序列的单碱基突变,并得出相对定量信息.本实验对不同浓度的HBV-DNA靶序列进行了检测.结果显示:此技术对单碱基突变有很强的特异性识别能力,并且具有较高的灵敏度(约10 pmol/L),在10～100 pmol/L浓度范围内表现出较好的线性关系.该技术检测时间短(＜1 h)、操作简单、不需要特殊的检测设备,具有很好的临床应用前景.     

Determination of Plasma Amino Acid Biomarkers by High Performance Liquid Chromatography for Diagnosis of Type 2 Diabetes Mellitus

The use of the fasting plasma glucose and 2-h plasma glucose tolerance tests leads to under-diagnosis of type 2 diabetes mellitus. To explore the contribution of metabolites for diagnosis for type 2 diabetes, plasma targeted metabolites (including free amino acids, lipids, and two hepatic aminotransferases) of 95 subjects were determined by high performance liquid chromatography and routine clinical methods. Receiver operating characteristic curve analysis was employed to evaluate potential diagnosis models. Based on the processing of orthogonal partial least squares-discriminant analysis, five amino acids (lysine, aspartate acid, threonine, methionine, and alanine) and two lipids (low density lipoprotein-cholesterol and high density lipoprotein-cholesterol) were discovered as potential biomarkers of type 2 diabetes. The area under the receiver operating characteristic curve built by the combination of these biomarkers and fasting plasma glucose was 0.994, which is significantly higher than that of the latter alone (0.907). These findings indicate that simultaneous measurement of specific metabolites with fasting plasma glucose from a single blood test for screening type 2 diabetes might be a more sensitive and specific strategy and provide additional insight regarding disease pathophysiologic mechanisms.     

单链脱氧核糖核酸探针在氧化锆陶瓷小珠表面的微阵列及陶瓷荧光小珠的制备

以N－（4－马来酰亚胺氧化丁酰）－琥珀酰亚胺磺酸钠盐（sulfo-GMBS)为介剂将5′－氨基／3′－得克萨斯红双末端修饰DNA探针偶联到3－（2－氨乙基氨丙基）三甲氧基硅烷（AATS）修饰的300μmZrO2陶瓷小珠表面，制成荧光小珠，小珠表面上单链DNA探针分子之间的距离取决于小球表面的预处理、偶联以应溶液中DNA探针浓度，小珠大小和小珠的数目。在优化条件下，20-mer单链DNA探针在小珠表面微阵列的最小分子间距约为7．6nm，即在一个300μmZrO2陶瓷小珠表面有10^10数量级的20－mer单链DNA探针参与微阵列反应。机理研究表明：DNA探针通过sulfo-GMBS偶联到小珠表面的化学反应仅涉及到探针末端修饰氨基。     

Bioactive Agent Discovery from the Natural Compounds for the Treatment of Type 2 Diabetes Rat Model

Diabetes mellitus is a well-known chronic metabolic disease that poses a long-term threat to human health and is characterized by a relative or absolute lack of insulin, resulting in hyperglycemia. Type 2 diabetes mellitus (T2DM) typically affects many metabolic pathways, resulting in β-cell dysfunction, insulin resistance, abnormal blood glucose levels, inflammatory processes, excessive oxidative reactions, and impaired lipid metabolism. It also leads to diabetes-related complications in many organ systems. Antidiabetic drugs have been approved for the treatment of hyperglycemia in T2DM; these are beneficial for glucose metabolism and promote weight loss, but have the risk of side effects, such as nausea or an upset stomach. A wide range of active components, derived from medicinal plants, such as alkaloids, flavonoids, polyphenol, quinones, and terpenoids may act as alternative sources of antidiabetic agents. They are usually attributed to improvements in pancreatic function by increasing insulin secretions or by reducing the intestinal absorption of glucose. Ease of availability, low cost, least undesirable side effects, and powerful pharmacological actions make plant-based preparations the key player of all available treatments. Based on the study of therapeutic reagents in the pathogenesis of humans, we use the appropriate animal models of T2DM to evaluate medicinal plant treatments. Many of the rat models have characteristics similar to those in humans and have the advantages of ease of genetic manipulation, a short breeding span, and access to physiological and invasive testing. In this review, we summarize the pathophysiological status of T2DM rat models and focus on several bioactive compounds from herbal medicine with different functional groups that exhibit therapeutic potential in the T2DM rat models, in turn, may guide future approach in treating diabetes with natural drugs.     

Dynamic Metabolic Profiling of Urine From Type 2 Diabetic KK-Ay Mice Treated with Repaglinide by GC-MS

Repaglinide is a short-acting insulin secretagogue, commonly used for the treatment of type 2 diabetes. In this paper, metabolomics were first applied to research of dynamic urine metabolic profiling and biomarkers of type 2 diabetic KK-A^y mice treated with repaglinide based on GC-MS. Twenty diabetic KK-A^y mice were randomly assigned to four groups and fed with repaglinide for 6, 9, 12, and 14 weeks, respectively. Five C57BL/6 J mice were used as the healthy control group and fed with water as contrast. The PCA scores plot of the identified 41 metabolites showed that as treating time went on, the diabetic groups got closer to the healthy group. Furthermore, five marker metabolites, d-Glucose, d-Galactose, 1,5-Anhydro-d-glucitol, myo-inositol and tartaric acid were screened out, which have similar change footprints of the whole metabolic profiles. The results demonstrated that repaglinide not only regulates the sugars and polyalcohol but also the organic acid in the organism. This work has illustrated the potential of metabolomics to disease diagnosis, pharmacology, and pharmacodynamics research.     

实时荧光聚合酶链式反应偶联高特异性核酸侵入反应检测单核苷酸多态性

建立了实时荧光聚合酶链式反应( PCR)偶联高特性核酸侵入反应检测单核苷酸多态性( SNP)的方法。优化了体系中flap核酸内切酶1(FEN1酶)和野生型检测探针等用量,确定了最佳反应条件,即FEN1酶用量为1.5 U,野生型检测探针用量为0.125μmol/L,0.5μmol/L Invader突变型检测探针,各0.25μmol/L通用野生型( VIC)和突变型( FAM)荧光共振转移发卡探针,显著降低了野生型样本和突变型样本背景信号,避免了背景信号对检测结果分型的干扰。采用本方法对编码乙醛脱氢酶2( ALDH2)基因ALDH2*2位点21例样本、细胞色素P4502C19基因CYP2C19*2和CYP2C19*3位点各19例样本进行分型检测,结果表明, AL-DH2*2位点GG纯合10例,GA杂合8例,AA纯合3例;CYP2C19*2位点GG纯合9例,GA杂合8例,AA纯合2例;CYP2C19*3位点GG纯合18例,GA杂合1例。使用焦磷酸测序进行验证,两种方法检测结果一致。本方法特异性好、操作简便、耗时短、成本低,可实现对SNP单管闭管无污染的分型检测。     

格列美脲治疗的2型糖尿病大鼠的尿液代谢组学研究

采用快速高分离度液相色谱-质谱技术(RRLC-MS)检测经格列美脲治疗的2型糖尿病大鼠尿液中代谢物的变化, 对糖尿病组、 给药组和健康对照组大鼠的尿液代谢物谱进行了分析. 采用主成分分析(PCA)对3组大鼠进行分类并寻找潜在生物标记物. 结果表明, 3组大鼠的尿液代谢物谱得到了很好的区分, 发现并鉴定了2个潜在生物标记物, 分别为4-脱氧三羟基丁酸和4-胍基丁酸. 格列美脲对2型糖尿病大鼠的药物作用可能体现为对氨基酸代谢的调节作用.     

Convenient synthesis of （2S,3R,4R,5S,6R）-2-（3-（4-ethylbenzyl）-4- chlorophenyl）-6-（hydroxymethyl）-tetrahydro- 2H-pyran-3,4,5-triol

A convenient approach for the preparation of （2S,3R,4R,5S,6R）-2-（3-（4-ethylbenzyl）-4-chlorophenyl）-6-（hydroxymethyl）- tetrahydro-2H-pyran-3,4,5-triol I is developed. The target compound via four steps is synthesized from 4-bromo-2-（bromomethyl）- 1-chlorobenzene and the isomers of undesired ortho-products were avoided during the preparation.     

快速高分辨液相色谱-质谱联用技术分析2型糖尿病大鼠口服四君子汤的药代动力学

研究四君子汤在2型糖尿病大鼠体内的药代动力学。 将四君子汤水提物以18 g/kg给正常大鼠和2型糖尿病大鼠灌胃,分别在不同时间点取尿液和粪便,检测其成分及代谢产物的含量;比较正常大鼠和2型糖尿病大鼠的药代动力学参数。 采用快速高分辨液相色谱-质谱联用技术对人参皂苷Rc、甘草甜素及其脱糖代谢物进行表征。 结果表明,四君子汤中人参皂苷Rc和甘草甜素在2型糖尿病大鼠体内的累计排泄率均高于正常大鼠,大部分人参皂苷通过尿液排出体外、粪便代谢为稀有人参皂苷;大部分甘草甜素在尿液和粪便中转化为甘草次酸,2型糖尿病大鼠与正常大鼠人参皂苷Rc和甘草甜素的药代动力学参数存在明显差异。 初步总结了四君子汤在体内的药代动力学机制和代谢途径。本分析方法具有高度自动化,灵敏性和准确性,这将有助于四君子汤的临床研究的不断发展。