Long-Lasting Expression of HO-1 Delays Progression of Type I Diabetes in NOD Mice

Heme oxygenase-1 (HO-1) is crucial in regulating oxidative injury. The present study was designed to assess whether HO-1 upregulation by cobalt protoporphyrin IX (CoPP) moderates or prevents the diabetic state in non-obese diabetic (NOD) mice, an animal model for Type 1 diabetes (T1D). HO-1 expression and HO activity were upregulated in the pancreas by the intermittent administration of CoPP. This was associated with decreases in blood glucose and pancreatic O2-, but increased pAKT and BcL-XL and cell survival. A considerable number of beta cells were preserved in the islets of CoPP-treated NOD mice, while none were found in untreated diabetic mice. The number of CD11c+ dendritic cells was decreased in the pancreas of CoPP-treated NOD mice (p     

移植后糖尿病与钙调神经蛋白免疫抑制剂使用的危险分析

目的:探讨移植后糖尿病(PTDM)的危险因素与钙调神经蛋白免疫抑制剂使用的关系.方法:分析近9年613例同种异体尸体肾移植术后患者年龄、性别、体质量指数(BMI)、糖尿病家族史、钙调神经蛋白免疫抑制剂(环孢霉A,CsA;他克莫司,FK506)使用与PTDM发生的危险程度.结果:PTDM的发生率为24.1%,PTDM发生与患者年龄、糖尿病家族史及BMI明显相关,PTDM患者钙调神经蛋白免疫抑制剂血药浓度剂量比明显高于移植后非糖尿病患者(non PTDM),且服用FK506后空腹血糖高于服用CsA.结论:与患者年龄、糖尿病家族史、BMI相比,服用钙调神经蛋白免疫抑制剂是导致PTDM发生最重要的危险因素,且FK506较CsA更易发生PTDM.     

Genetic and Molecular Basis of QTL of Diabetes in Mouse: Genes and Polymorphisms

A systematic study has been conducted of all available reports in PubMed and OMIM (Online Mendelian Inheritance in Man) to examine the genetic and molecular basis of quantitative genetic loci (QTL) of diabetes with the main focus on genes and polymorphisms. The major question is, What can the QTL tell us? Specifically, we want to know whether those genome regions differ from other regions in terms of genes relevant to diabetes. Which genes are within those QTL regions, and, among them, which genes have already been linked to diabetes? whether more polymorphisms have been associated with diabetes in the QTL regions than in the non-QTL regions.Our search revealed a total of 9038 genes from 26 type 1 diabetes QTL, which cover 667,096,006 bp of the mouse genomic sequence. On one hand, a large number of candidate genes are in each of these QTL; on the other hand, we found that some obvious candidate genes of QTL have not yet been investigated. Thus, the comprehensive search of candidate genes for known QTL may provide unexpected benefit for identifying QTL genes for diabetes. Key Words: Quantitative trait loci, type 1 diabetes, insulin-dependent diabetes mellitus (IDDM), candidate gene, polymorphism, mouse.     

Prevention of Cyclophosphamide-induced Accelerated Diabetes in the NOD Mouse by Nicotinamide or a Soy Protein-based Infant Formula

Spontaneous diabetes in the NOD mouse can be prevented by nicotinamide or by an infant formula diet in which the protein source is replaced with casein hydrolysate (Pregestimil) or soy protein (Prosobee). NOD mice maintained on the standard diet (chow and water) and given cyclophosphamide (Cy) at day 95 develop accelerated and synchronised diabetes within 14 days. Here, we compared the ability of oral nicotinamide or Prosobee, either given alone or concurrently, from weaning, in preventing diabetes in the Cy model. The resulting insulitis and the expression of intra-islet inducible nitric oxide synthase (iNOS) were examined at days 0, 4, 7, 11 and 14 following Cy administration. Intra-islet CD4 and CD8 cells and macrophages were also enumerated at day 11. In mice given the standard diet and injected with Cy at day 95 (group 5), diabetes developed in 7/11 mice, 14 days later. Mice exposed to oral nicotinamide (group 2), Prosobee (group 3) or both (group 4), did not develop the disease during this period and until a further 30 days (p = 0.03). In mice exposed to the standard diet and without Cy treatment (group 1) the insulitis scores increased slowly until day 11 and then declined slightly at day 14 whereas mice exposed to the same diet but given Cy at day 95, showed a sharp decline at day 4 followed by a rapid increase between day 7–14. However, in mice given either nicotinamide, Prosobee or both, the insulitis scores at most time-points were generally lower than in Cy-teated animals on the standard diet. In the latter group, CD4 and CD8 cells and macrophages were also higher at day 11 than all other 4 groups (CD4: p < 0.05; CD8: p < 0.05; macrophages: p < 0.0001). The number of iNOS labelled cells increased progressively in mice on the standard diet and given Cy and were significantly higher at days 4, 7 and 11 than in the 3 dietary groups. Thus, oral nicotinamide or Prosobee, either alone or together, prevents Cy induced diabetes in the NOD mouse. The protective diets suppress Cyinduced intra-islet immune cell influx and iNOS expression.     

Liver Grafts for Transplantation from Donors with Diabetes: An Analysis of the Scientific Registry of Transplant Recipients Database

Patients with a history of diabetes mellitus (DM) have worse survival than those without DM after liver transplantation. However, the effect of liver grafts from DM donors on the post-transplantation survival of recipients is unclear. Using the Scientific Registry of Transplant Recipients database (2004–2008), 25,413 patients were assessed. Among them, 2,469 recipients received grafts from donors with DM. The demographics and outcome of patients were assessed. Patient survival was assessed using Kaplan–Meier methodology and Cox regression analyses. Recipients from DM donors experienced worse graft survival than recipients from non-DM donors (one-year survival: 81% versus 85%, and five-year survival: 67% versus 74%, P<0.001, respectively). Graft survival was significantly lower for recipients from DM donors with DM duration >5 years (P<0.001) compared with those with DM duration <5 years. Cox regression analyses showed that DM donors were independently associated with worse graft survival (hazard ratio, 1.11; 95% confidence interval, 1.02–1.19). The effect of DM donors was more pronounced on certain underlying liver diseases of recipients. Increases in the risk of graft loss were noted among recipients from DM donors with hepatitis-C virus (HCV) infection, whereas those without HCV experienced similar outcomes compared with recipients from non-DM donors. These data suggest that recipients from DM donors experience significantly worse patient survival after liver transplantation. However, in patients without HCV infection, using DM donors was not independently associated with worse post-transplantation graft survival. Matching these DM donors to recipients without HCV may be safe.     

Diabetes Increases the Expression of Hypothalamic Neuropeptides in a Spontaneous Model of Type I Diabetes, the Nonobese Diabetic (NOD) Mouse

1. Synthesis of oxytocin (OT) and arginine-vasopressin(AVP) is increased in induced models of Type I diabetes, such as thestreptozotocin model. However, these parameters have not yet been evaluated inspontaneous models, such as the nonobese diabetic mouse (NOD). Therefore, we studied in the magnocellular cells of the paraventricular nucleus (PVN)of nondiabetic and diabetic 16-week-old female NOD mice and control C57Bl/6mice, the immunocytochemistry of OT and AVP peptides and their mRNA expression, using nonisotopic in situ hybridization (ISH).2. In nondiabetic and diabetic NOD female mice, the number of OT- and AVP-immunoreactive cells were similar to those of the controls, whereas immunoreaction intensity was significantly higher for both peptides in diabetic NOD as compared with nondiabetic NOD and control C57Bl/6 mice.3. ISH analysis showed that the number of OT mRNA-containing cells was in the same range in the three groups, whereas higher number of AVP mRNA expressing cells was found in diabetic NOD mice. However, the intensity of hybridization signal was also higher for both OT and AVP mRNA in the diabetic group as compared with nondiabetic NOD and control mice.4. Blood chemistry demonstrated that haematrocrit, total plasma proteins, urea, sodium, and potassium were within normal limits in diabetic mice. Thus, NODmice were neither hypernatremic nor dehydrated.5. We suggest that upregulation of OT and AVP reflects a high-stress condition in the NOD mice. Diabetes may affect neuropeptide-producing cells of the PVN, with the increased AVP and OT playing a deleterious role on the outcome of the disease.     

Prevention of Diabetes in the NOD Mouse by Intra-muscular Injection of Recombinant Adeno-associated Virus Containing the Preproinsulin II Gene

Using the Adeno-associated virus (AAV) as a gene delivery vehicle, we have constructed a recombinant vector containing the full length rat preproinsulin gene (vLP-1). Utilizing the well described non-obese diabetic (NOD) mouse model, an experimental group (n=10) of animals were intramuscularly (IM) injected with 10⁷ rAAV virions containing the insulin gene and compared to a mock-injected control group (n=10). Blood glucose (glc) was then measured weekly for 16 weeks. Data showed that the experimental group contained 70% euglycemic animals (defined as glc <200mg/dL) versus 10% of the control animals (P<.05) at 14 weeks. Mean weight in the treated group was greater than the untreated group. Insulin mRNA was detected at the injection site of all of the treated animals, but not controls. Complete destruction of islets was confirmed by histology ruling out the possibility of spontaneous reversal of insulinitis. We conclude that IM delivery of the insulin gene in the NOD mouse was able to prevent clinical DM up to 14 weeks in a majority of treated animals. Our experimental data suggests that gene therapy may be an alternative treatment for IDDM in the future.     

小鼠胚胎紧密化现象分子基础的研究进展

就小鼠植入前胚胎紧密化过程中胚胎的形态变化、细胞极化、细胞间连接及紧密化的物质基础、时控机制等作了综述.对紧密化现象的分子基础研究大多数都集中在翻译后水平的调控,而在基因水平的研究较少.     

Tfh 细胞在NOD小鼠糖尿病发病过程中的作用机制的研究

目的:研究滤泡辅助性T细胞(Follicular Helper T cell,Tfh)在非肥胖性糖尿病小鼠(Non-obese Diabetic mice,NOD)发病过程中的作用机制。方法:实验动物NOD小鼠按血糖值分为胰岛炎组(血糖浓度≤9 mmol/L)及糖尿病组(血糖浓度≥20 mmol/L)。ELISA法检测各组中糖尿病自身抗体谷氨酸脱羧酶抗体(65-kda glutamate decarboxylase antibody,GAD65Ab)、抗胰岛素自身抗体(Insulin autoantibody,IAA)表达水平,Western blot检测B细胞型淋巴瘤6蛋白(B-cell lymphoma 6 protein,Bcl-6)及可诱导共刺激分子(Inducible costimulatory molecule,ICOS)表达,流式细胞仪检测各组外周血及脾脏Tfh细胞水平。结果:糖尿病组NOD鼠自身抗体GAD65Ab(1.21±0.23 nmol/L)、IAA(0.96±0.12 nmol/L)浓度较胰岛炎组(0.32±0.09 nmol/L,0.25±0.06 nmol/L)均有明显升高;糖尿病组NOD鼠Bcl-6及ICOS表达较胰岛炎组NOD鼠有明显升高,外周血和脾脏Tfh细胞水平糖尿病组NOD鼠(24.55%)较胰岛炎组NOD鼠(4.27%)升高明显。结论:NOD小鼠自发糖尿病与自身抗体浓度升高相关,Tfh细胞可能参与NOD鼠糖尿病发生及发展过程。     

A Mouse Model of in Utero Transplantation

The transplantation of stem cells and viruses in utero has tremendous potential for treating congenital disorders in the human fetus. For example, in utero transplantation (IUT) of hematopoietic stem cells has been used to successfully treat patients with severe combined immunodeficiency.^1,2 In several other conditions, however, IUT has been attempted without success.³ Given these mixed results, the availability of an efficient non-human model to study the biological sequelae of stem cell transplantation and gene therapy is critical to advance this field. We and others have used the mouse model of IUT to study factors affecting successful engraftment of in utero transplanted hematopoietic stem cells in both wild-type mice^4-7 and those with genetic diseases.^8,9 The fetal environment also offers considerable advantages for the success of in utero gene therapy. For example, the delivery of adenoviral¹⁰, adeno-associated viral¹⁰, retroviral¹¹, and lentiviral vectors^12,13 into the fetus has resulted in the transduction of multiple organs distant from the site of injection with long-term gene expression. in utero gene therapy may therefore be considered as a possible treatment strategy for single gene disorders such as muscular dystrophy or cystic fibrosis. Another potential advantage of IUT is the ability to induce immune tolerance to a specific antigen. As seen in mice with hemophilia, the introduction of Factor IX early in development results in tolerance to this protein.¹⁴In addition to its use in investigating potential human therapies, the mouse model of IUT can be a powerful tool to study basic questions in developmental and stem cell biology. For example, one can deliver various small molecules to induce or inhibit specific gene expression at defined gestational stages and manipulate developmental pathways. The impact of these alterations can be assessed at various timepoints after the initial transplantation. Furthermore, one can transplant pluripotent or lineage specific progenitor cells into the fetal environment to study stem cell differentiation in a non-irradiated and unperturbed host environment.The mouse model of IUT has already provided numerous insights within the fields of immunology, and developmental and stem cell biology. In this video-based protocol, we describe a step-by-step approach to performing IUT in mouse fetuses and outline the critical steps and potential pitfalls of this technique.Download video file.^{(55M, mov)}     

Inhibition of Autoimmune Diabetes in NOD Mice by miRNA Therapy

Autoimmune destruction of the pancreatic islets in Type 1 diabetes is mediated by both increased proinflammatory (Teff) and decreased regulatory (Treg) T lymphocytes resulting in a significant decrease in the Treg:Teff ratio. The non-obese diabetic (NOD) mouse is an excellent in vivo model for testing potential therapeutics for attenuating the decrease in the Treg:Teff ratio and inhibiting disease pathogenesis. Here we show for the first time that a bioreactor manufactured therapeutic consisting of a complex of miRNA species (denoted as TA1) can effectively reset the NOD immune system from a proinflammatory to a tolerogenic state thus preventing or delaying autoimmune diabetes. Treatment of NOD mice with TA1 resulted in a systemic broad-spectrum upregulation of tolerogenic T cell subsets with a parallel downregulation of Teff subsets yielding a dramatic increase in the Treg:Teff ratio. Moreover, the murine-derived TA1 was highly effective in the inhibition of allorecognition of HLA-disparate human PBMC. TA1 demonstrated dose-responsiveness and exhibited equivalent or better inhibition of allorecognition driven proliferation than etanercept (a soluble TNF receptor). These findings demonstrate that miRNA-based therapeutics can effectively attenuate or arrest autoimmune disease processes and may be of significant utility in a broad range of autoimmune diseases including Type 1 diabetes.     

NOD/SCID小鼠模型在实验血液学研究中的应用

NOD/SCID（非肥胖糖尿病/重症联合免疫缺陷）小鼠是在SCID（重症联合免疫缺陷）小鼠的基础上与非肥胖性糖尿病小鼠（NOD/Lt）品系回交的免疫缺陷鼠。NOD/SCID小鼠既有先天免疫缺陷,又有T和B淋巴细胞缺乏,各种肿瘤细胞可以植入,且较少发生排斥反应及移植物抗宿主病（GVHD）,所以NOD/SCID小鼠逐渐成为血液学实验研究的有用工具。本文从NOD/SCID小鼠的生物学特性、建立人类白血病模型、干细胞移植、药物研究及NOD/SCID小鼠应用中存在的不足和改良等方面综合述评。     

免疫耐受在胰岛移植中的研究进展

胰岛移植已经被公认为治疗胰岛素依赖型糖尿病（IMDD）的有效手段,而现如今胰岛移植的最大障碍是移植排斥反应。目前控制胰岛移植的免疫抑制治疗因其对胰岛细胞的毒性作用及长期应用带来的全身并发症而无法在临床推广,诱导移植术后受体的免疫耐受是防止排斥反应的最理想方法。本文综述了诱导免疫耐受的途径及胰岛移植的最新实验进展。随着研究的深入和免疫学的发展,相信在未来的胰岛移植治疗糖尿病领域,移植排斥现象将能得到高效可靠的解决。     

免疫耐受在胰岛移植中的研究进展

胰岛移植已经被公认为治疗胰岛素依赖型糖尿病(IMDD)的有效手段,而现如今胰岛移植的最大障碍是移植排斥反应。目前控制胰岛移植的免疫抑制治疗因其对胰岛细胞的毒性作用及长期应用带来的全身并发症而无法在临床推广,诱导移植术后受体的免疫耐受是防止排斥反应的最理想方法。本文综述了诱导免疫耐受的途径及胰岛移植的最新实验进展。随着研究的深入和免疫学的发展,相信在未来的胰岛移植治疗糖尿病领域,移植排斥现象将能得到高效可靠的解决。     

胚胎干细胞移植治疗糖尿病的研究进展

糖尿病是一种严重的免疫缺陷性疾病,目前的治疗方法很难从根本上治愈。近年来的研究表明,通过诱导胚胎干细胞定向分化为胰岛β细胞,并进行移植治疗糖尿病,是一种有希望的根治方案。本文就利用胚胎干细胞移植治疗糖尿病的最新进展作一综述。     

Effect of Statins on the Progression of Coronary Calcification in Kidney Transplant Recipients

BackgroundCoronary calcification (CAC) is highly prevalent in kidney transplant recipients (KTRs) and has been associated with cardiovascular morbidity and mortality. Some studies have shown a reduction in CAC progression with statin therapy in the general and chronic kidney disease (CKD) populations.ResultsAt baseline, CAC was observed in 30% and 21% of patients in the statin and control groups, respectively (p = 0.39). The calcium score at baseline and its absolute and relative changes over 12 months of follow up were similar among the groups. In the statin group, total cholesterol (p < 0.001), low density lipoprotein cholesterol (p < 0.001) and triglycerides (p = 0.005) decreased, and the estimated glomerular function rate increased (p<0.001) significantly. CRP levels remained stable (p = 0.52) in the statin group but increased in the control group (p = 0.01). In the multivariate model, there was no difference in CAC progression between the groups (group effect p = 0.034; time-effect p = 0.23; interaction p = 0.74). Similar results were obtained when only patients with ≥ 10AU calcium score (calcified) were analyzed (group effect p = 0.051; time-effect p = 0.58; interaction p = 0.99).ConclusionAlthough statins reduce the levels of cholesterol, triglycerides, inflammation and improve graft function, the dose adopted in the current study did not delay CAC progression within 12 months of follow up.

Trial Registration

Brazilian Clinical Trials Registry RBR-32RFMB     

Predictors of Diabetes in Older People in Urban China

Background

China has the largest number of people with diabetes in the world. Over the last 30 years China has experienced rapid economic growth and a growing income gap between rich and poor. The population is ageing, however diabetes in older people has not been well studied to date. In this study we determined incidence and predictors of diabetes in older Chinese people.

Methods

During 2001, using a standard interview method, we examined 1,317 adults aged ≥65 years who did not have diabetes in the city of Hefei, and characterized baseline risk factors. Over 7.5 years of follow up, we documented incident diabetes using self-reported doctor diagnosis and the cause of death in the whole cohort, and HbA_1C ≥48 mmol/mol in a nested case-control sample. A multivariate Cox regression model was employed to investigate risk of diabetes in relation to baseline risk factors.

Results

During follow up, 119 persons had newly diagnosed diabetes. World age-standardised incidence of diabetes was 24.5 (95% CI 19.5–29.5) per 1,000 person-years. Risk of diabetes was significantly and positively associated with income, waist circumference and body mass index, smoking and uncontrolled hypertension, but negatively associated with having a hobby of walking and frequency of visiting children/other relatives and contacting neighbours/friends. Higher income was significantly associated with increased diabetes risk regardless of cardiovascular and psychosocial risk factors. Compared to those with middle income and no psychosocial risk factors, the hazard ratio for incident diabetes among participants with high income and psychosocial risk was 2.13 (95% CI 1.02–4.45).

Conclusions

Increasing incidence of diabetes in relation to high income has become an important public health issue in China. Maintaining social networks and gentle physical activities and reducing psychosocial factors may be integrated into current multi-faceted preventive strategies for curbing the epidemic of diabetes in the older population.