Remission in models of type 1 diabetes by gene therapy using a single-chain insulin analogue

A cure for diabetes has long been sought using several different approaches, including islet transplantation, regeneration of beta cells and insulin gene therapy. However, permanent remission of type 1 diabetes has not yet been satisfactorily achieved. The development of type 1 diabetes results from the almost total destruction of insulin-producing pancreatic beta cells by autoimmune responses specific to beta cells. Standard insulin therapy may not maintain blood glucose concentrations within the relatively narrow range that occurs in the presence of normal pancreatic beta cells. We used a recombinant adeno-associated virus (rAAV) that expresses a single-chain insulin analogue (SIA), which possesses biologically active insulin activity without enzymatic conversion, under the control of hepatocyte-specific L-type pyruvate kinase (LPK) promoter, which regulates SIA expression in response to blood glucose levels. Here we show that SIA produced from the gene construct rAAV-LPK-SIA caused remission of diabetes in streptozotocin-induced diabetic rats and autoimmune diabetic mice for a prolonged time without any apparent side effects. This new SIA gene therapy may have potential therapeutic value for the cure of autoimmune diabetes in humans.     

电针和促进剂对胰岛素口腔黏膜渗透的影响

研究在电针仪作用下不同吸收促进剂对胰岛素口腔黏膜渗透性的影响.测定在使用电针仪前后不同吸收促进剂对胰岛素透过家兔口腔黏膜的渗透系数及血糖降低情况.加入吸收促进剂可以增加胰岛素透过口腔黏膜的渗透作用,使用电针仪后渗透作用显著提高.电针仪可协助吸收促进剂提高胰岛素口腔黏膜吸收.     

Recombinant Expression of the Monomeric B27 Lys Destripeptide Insulin Precursor( MIP) in Escherichia coli

Insulin controls hyperglycemia caused by diabetes,and virtually all treatments require exogenous insulin. The monomeric B27 Lys destripeptide insulin has 80% bioactivity of wild type. It is a potential drug for clinic. But the yield of monomeric B27 Lys destripeptide insulin is limited, because monomeric B27 Lys destripeptide insulin precursor( MIP) was easily to form inclusion body,when MIP is expressed in E. coli. The precursor,MIP,is not only fused to an N-terminal HIS-small ubiquitin-related modifier( SUMO) tag,but also added five Arginine( 5 R) at the C-terminal,which can improve the solubility of the fusion protein. By this way,the yield of fusion protein has been up to 1. 45 mg/L cell culture.This work set up the foundation for the clinical application of B27 Lys destripeptide insulin.     

Effects of breakfast with different calorigenic amounts on blood glucose, insulin and glucagon levels

This study was aimed to investigate the relationship between breakfast and serum glucose, insulin and glucagon concentrations in order to establish a model breakfast appropriate for Chinese. Twenty-four volunteers were randomly assigned to four study groups: high carbohydrate breakfast, high fat and protein breakfast, the typical breakfast and fasting. Each subject had serum and urine samples collected while fasting and at 1,2 and 3.5 hours following the meal. The concentration of serum glucose, insulin and glucagon was measured. The levels of serum glucose in group A, B and C differed significantly at 1 and 2 hour after meal compared to those at fasting (P<0.05). The serum glucose in group A increased insignificantly after meal. The serum insulin levels were in group A, B and C significant different compared with control group(P<0.05).Those peaked at 1 hour after meal, with group C rising the furthest. Compared with the fasting group, the serum glucagons rose and maintained the increase after breakfast in group A, B and C (P<0.05). The data suggested that various diets with different calorigenic amounts increased hormone concentration to various extents. We found that a breakfast rich in carbohydrates could maintain proper blood glucose level.     

胰岛素联合口服药二甲双胍治疗2型糖尿病的临床观察

目的：观察早、晚皮下注射预混胰岛素联合口服二甲双胍治疗2型糖尿病的临床效果．方法：对60例早、晚注射预混胰岛素治疗效果欠佳的2型糖尿病患者加用口服降糖药,分别测定治疗3个月前后的空腹血糖（FPG）、餐后2小时血糖（2hPG）、糖化血红蛋白（HbAlc）、空腹C肽、餐后2小时C肽、血脂、体重、血压．结果：加用口服药治疗后患者的空腹血糖、餐后2小时血糖、糖化血红蛋白、体重指数较治疗前明显下降（P〈0．01）,餐后2小时C肽较治疗前明显升高,其差异有显著性（P〈0．01）,而对血压、血脂影响不大．结论：对单用胰岛素治疗效果欠佳的2型糖尿病联合应用二甲双胍治疗效果显著．     

Lanthanides' enhancing absorption of insulin and reduction of blood glucose of rat by pulmonary administration

To explore the possibility of lanthanides as the enhancer of insulin absorption, the promoting effects of lanthanide ions on the absorption of intrapulmonary delivered insulin and reduction of blood glucose were investigated by means of two approaches, preadministration and coadministration of lanthanide ions with insulin. The results indicate that, compared with the results of those given insulin only, lanthanide ions can effectively enhance the level of insulin and the reduction of glucose level in blood. These effects are dependent on lanthanide species and their concentrations. The mechanism was discussed.     

Lanthanides’ enhancing absorption of insulin and reduction of blood glucose of rat by pulmonary administration

To explore the possibility of lanthanides as the enhancer of insulin absorption, the promoting effects of lanthanide ions on the absorption of intrapulmonary delivered insulin and reduction of blood glucose were investigated by means of two approaches, preadministration and coadministration of lanthanide ions with insulin. The results indicate that, compared with the results of those given insulin only, lanthanide ions can effectively enhance the level of insulin and the reduction of glucose level in blood. These effects are dependent on lanthanide species and their concentrations. The mechanism was discussed.     

胰岛素类似物Aspart应用于胰岛素泵治疗2型糖尿病急性并发症疗效分析

比较了Aspart与短效可溶性胰岛素对高渗性非酮症糖尿病昏迷(hyperosmolar dia-betic nonketotic coma HONK)或酮症酸中毒(diabetic ketoacidosis DKA)的治疗差异。发现胰岛素泵持续皮下注射治疗组和短效可溶性胰岛素(诺和灵R)对照组间年龄、血糖和体重指数(BMI)无显著性差异;观察血糖、血酮、尿酮体及渗透压、血钠的变化,低血糖的发生率,结果表明:(1)两组患者的血糖经治疗后均明显下降,但达标血糖值的时间治疗组较对照组缩短,药物用量较对照组减少,差异较显著;(2)两组患者血酮恢复正常时间、尿酮体转阴时间、血气pH值恢复正常时间与对照组相比均无显著性差异(P>0.05);(3)治疗组低血糖发生率〔(0.32)次/人〕较对照组〔(1.7)次/人〕低,差异有显著意义(P<0.01)。胰岛素类似物Aspart胰岛素泵持续皮下注射治疗DKA、HONK同短效可溶性胰岛素胰岛素泵持续皮下注射均能取得较好疗效,但前者更有效,安全性更好。     

Three mutant insulins in man

We have previously identified a structurally abnormal insulin in the serum and pancreas of a middle-aged man with diabetes mellitus which arose from a leucine for phenylalanine substitution at position 24 or 25 of the insulin B chain; further analysis of the patient's leukocyte DNA showed that one of the patient's insulin alleles had undergone mutation resulting in loss of an MboII restriction site normally present in the human insulin gene. Two additional and unrelated patients with the same clinical syndrome have now been identified (ref. 4 and unpublished results). All of these patients showed hyperglycaemia typical of diabetes and with marked hyperinsulinaemia typical of insulin resistance, but all three show normal tolerance to exogenously administered insulin. As the opportunity of examining pancreatic tissue from patients suspected of secreting insulin variants is rare, we have developed a method combining HPLC and radioimmunoassay to identify insulin variants isolated from human sera. By this method we have shown that all three patients noted above secrete structurally variant and chemically distinct insulins. In correction of our original assignment, one is identified as [LeuB25]insulin.     

Evolutionary change in the insulin receptors of hystricomorph rodents

INSULIN receptors have been thought to have remained unaltered despite evolutionary changes in the hormone(1). Because insulins from hystricomorph rodents are known to be highly substituted compared with other mammalian insulins, we decided to investigate the insulin receptor of some hystricomorphs to determine whether evolutionary change had occurred within the receptor itself. Here we present the first evidence that hystricomorph rodent insulin receptors have undergone evolutionary change.     

某底排弹底排装置工作期间内部流场的数值模拟

目的:探讨动态血糖监测系统(CGMS)联合胰岛素泵持续皮下输注胰岛素(CSII)治疗初诊重症2型糖尿病(T2DM)的有效性及安全性.方法:48例初诊重症T2DM病人分为试验组与对照组,试验组CGMS与CSII联合应用,当皮下探头信号衰减时改为每日7次指尖血糖监测指导CSII直至血糖达标;对照组始终用每日7次指尖血糖监测指导CSII血糖达标.观察两组血糖达标所需天数、全天胰岛素用量、低血糖发生率和各时间点血糖来评价治疗的有效性及安全性.结果:试验组血糖达标所需天数、全天胰岛素用量、低血糖发生率和各时间点血糖均低于对照组.结论:试验组治疗初诊重症T2DM有效性及安全性优于对照组.     

心理干预对维持性血液透析患者影响的随机对照研究

将77例维持性血液透析患者随机分为干预组或对照组。对照组接受常规的血液透析并按医嘱服用重组人促红细胞生成素（rhEPO）;干预组在此基础上,还接受渐进性肌肉放松训练和脑波同步治疗。采用SCL-90量表和SF-36问卷,在干预前、后3天内分别对两组进行测量,同时检测血常规、肾功能及透析间期体重增加量与干体重的比值(IWGR%)。70例患者（干预组36例,对照组34例）完成了实验。结果发现,经过心理干预,干预组的心理健康状况、总体生活质量、血红蛋白（HB）、红细胞计数（RBC）、红细胞压积（HCT）以及透析前血清钙(Ca²⁺)显著高于对照组;透析前血清尿素氮（BUN）、磷 (PO^3-₄)、钾（K⁺）水平以及透析间期体重增加量与干体重的比值（IWGR%）显著低于对照组;两组透析前的血清肌酐（Scr）差异无显著性意义。     

天然矿泉水与纯净水抗过氧化作用的比较

将6周龄纯种BALB/C小鼠随机分为4组,一组为皮下注射生理盐水的非致衰对照组,另三个组为注射D-半乳糖的致衰模型组,它们分别饮用天然矿泉水,纯净水和自来水.4个月后,测定红细胞超氧化物歧化酶(SOD)和全血谷胱甘肽过氧化物酶(GSH-Px)的活性.本实验表明,天然矿泉水具有一定的抗过氧化作用.天然矿泉水组与自来水致衰对照组比较,GSH-Px活力变化不显著,而SOD活力明显提高( 7.1%).而纯净水组与天然矿泉水组比较,SOD活力降低了10.2%,GSH-Px活力降低15.5%,两项指标均有显著差异.值得注意的是,纯净水组与自来水致衰对照组相比较,其抗过氧化作用也有所减弱,纯净水组的SOD活力下降3.9%,GSH-Px活力更是显著地下降了18.5%.     

糖尿病患者血糖控制不佳时治疗方案选择的参考指标

目的探讨糖尿病患者血糖控制不佳时治疗方案选择的参考指标.方法选择血糖控制不佳的312例Ⅱ型糖尿病患者,入院后行胰岛素强化降糖治疗,血糖稳定后检测胰岛功能,空腹C肽≥1 ng/m L的患者改为口服促进胰岛素分泌类药物联合基础胰岛素治疗(方案1),空腹C肽1 ng/m L的患者继续胰岛素强化降糖治疗,4次/d(方案2);接受方案1的患者连续治疗7 d后若空腹血糖和餐后2 h血糖未达标,则改行方案2.比较两组患者性别、年龄、体质量指数、糖尿病病程、空腹血糖、糖化血红蛋白、餐后2 h血糖、空腹C肽、餐后2 h C肽水平.结果两组患者性别、年龄、糖尿病病程之间差异均无统计学意义(P0.05);方案1组患者体质量指数、空腹C肽、餐后2 h C肽、Δ空腹C肽明显高于方案2组,空腹血糖、餐后2 h血糖、糖化血红蛋白明显低于方案2组,差异均具有统计学意义(P0.05).经胰岛素强化治疗血糖稳定后分组,两组患者胰岛素剂量、低血糖发生率、空腹血糖达标率、餐后2 h血糖达标率之间差异无统计学意义(P0.05);治疗30 d门诊随访,方案1组胰岛素剂量、低血糖发生率明显低于方案2组,空腹血糖达标率、餐后2 h血糖达标率明显高于方案2组,差异具有统计学意义(P0.05).二元逐步Logistic回归分析显示:体质量指数、Δ空腹C肽为选择方案1的独立影响因素.结论糖尿病患者血糖控制不佳时,体质量指数、餐后2 h C肽可作为降糖方案选择的预测指标.     

A simple, economical method of converting gene expression products of insulin into recombinant insulin and its application

A method, by which the gene expression product of recombinant single chain insulin can be converted into insulin by directly digesting with trypsin, has been established. This method has been used in process of porcine insulin precursor (PIP), [B16Ala]PIP and [B26Ala]PIP into (desB30)insulin, (desB30)[B16Ala]insulin and (desB30)[B26Ala]insulin, respectively, and all of them retain full biological activity of that of their corresponding parent, recombinant human insulin, [B16Ala]insulin and [B26Ala]insulin. The results further demonstrate that the C-terminal residue of B chain is not necessary for insulin's biological activity. Compared with the method of transpeptidation, this method is simple, with a high yield, and avoids the use of organic reagents, and in comparison with the trypsin/carboxypeptidase method, it omits the use of carboxylpeptidase. Besides, (desB30)[B16Ala]insulin and (desB30)[B26Ala]insulin still remained without self-association property as that of their parents, which demonstrate that they are monomeric insulin. So they can be used for substituting for monomeric insulin, [B16Ala]insulin and [B26Ala]insulin, in clinical applications.     

生理溶液中磺化酞菁溶液状态的研究

水溶性磺化酞菁能选择性地滞留在肿瘤组织中，是一种新的光动力治疗肿瘤的敏化剂。有关酞菁化合物光动力治疗肿瘤的机制到目前还不清楚，而在生理条件下光敏剂的状态研究对阐明其机制有重要意义。在生理条件下（ｐＨ＝７．２， ０． １５ ｍｏｌ／Ｌ ＮａＣｌ水溶液）对合成的 ３种磁化酞菁（ＺｎＰｃＳ，ＣｌＡｌＰｃＳ，Ｈ２ＰｃＳ）进行吸收光谱研究，探讨了它们在溶液中的聚集状态。结果是ＣｌＡｌＰｃＳ在实验的条件下不发生聚集，而其他两种都呈聚集态。通过分析为二聚体，并获得了平衡常数和单体摩尔吸收系数。     

补肾活血颗粒对多囊卵巢综合征大鼠血清性激素水平、 血糖及胰岛素的影响

目的 观察补肾活血颗粒对多囊卵巢综合征（PCOS）模型大鼠血清性激素水平、血糖及胰岛素的影响。方法 采用颈背部皮下注射脱氢表雄酮（DHEA）造模法建立PCOS大鼠模型,将72只雌性SD大鼠随机分为正常对照组、模型组、二甲双胍组、中药高剂量组、中药中剂量组、中药低剂量组,每组12只,各组给予相应药物连续灌胃 20 d,用酶联免疫吸附测定法测定血清17-羟孕酮（17-OHP）、促黄体生成素（LH）、睾酮（T）、促卵泡素（FSH）、空腹 血糖（FPG）及胰岛素（INS）水平。结果模型组血清中17-OHP、LH、T、FSH、FPG、INS水平明显高于正常对照组 (P<0.01 ,P<0. 05 ),中药高剂量组降低大鼠血清LH、T、FSH及INS水平最为明显(P<0.01),二甲双胍组、中药中剂量组显著降低FPG（P<0. 01）,中药各治疗组均能降低17-OHP（P<0. 05）。结论 补肾活血颗粒能对PCOS 大鼠有一定的治疗作用,可改善激素水平、血糖及胰岛素的异常     

罗格列酮对老年2型糖尿病合并高血压患者血压的影响

目的:观察胰岛素增敏剂罗格列酮用于老年2型糖尿病合并高血压的患者时,降低血浆胰岛素水平以及改善胰岛素敏感性后对血压的影响。方法:23例老年2型糖尿病合并高血压患者口服罗格列酮(文迪雅)4mg～8mg,1次/日。治疗前后测定BMI、血压、HbA_(1C)、FBG、PBG、FINS、PINS,并按HOMA模型计算胰岛素抵抗指数、胰岛素分泌指数和药物的不良反应。结果:(1)罗格列酮治疗12周后,BMI治疗前后比较无显著性差异;血压治疗后明显下降,降压幅度达(6.75±0.03)/(7.25±0.53)mmHg,与治疗前比较有显著性差异。(2)治疗后HbA_(1C)、FBG、PBG、FINS、PINS均明显下降,治疗前后比较有显著性差异。(3)治疗后HOMA-IR下降,IAI升高,与治疗前比较有显著性差异,FINS/FBG无显著性差异。(4)我国老年患者使用罗格列酮的安全性及耐受性较好。结论:罗格列酮治疗中国老年2型糖尿病患者,能降低血浆胰岛素水平,改善IR,显著降低收缩压和舒张压;且其安全性、耐受性均较好。     

Growth restoration of insulin-deficient diabetic rats by recombinant human insulin-like growth factor I

Insulin-like growth factor I (IGF-I) and insulin stem from a common precursor, are structural homologues, act through similar receptors and elicit insulin-like and growth-promoting effects in vitro and in vivo. Serum IGF-I levels are controlled by growth hormone, insulin and nutrition. Insulin-deficient growth-arrested diabetic animals have reduced serum IGF-I levels which are restored towards normal by insulin but not by growth-hormone treatment. Here we show that normal growth of diabetic rate is restored by infusion of recombinant human (rh)IGF-I without normalization of the blood sugar level and that insulin acts via an increase of IGF-I synthesis on growth of diabetic rats. We describe a new mechanism of endocrine control of growth in which IGF-I is the major stimulator at the cellular level. Growth hormone and insulin act mainly by modulating the hepatic synthesis of IGF-I.     

Receptor-binding region of insulin.

X-ray analysis, circular dichroism, receptor binding and biological potencies of chemically modified insulins suggest that the conformation of the insulin molecule is critical to the formation of both the zinc insulin hexamer and the insulin-receptor complex. Results are consistent with an insulin receptor-binding region including many of the hydrophobic residues important to dimerisation in addition to more polar surface residues. There is a further possibility of formation of an antiparallel sheet structure between the insulin and receptor molecules in the complex similar to that between monomers in the insulin dimer.