Changes of Regulatory T Cells in the Early Stage of Obesity Mice and Their Modulation on Macrophage Subtypes in Visceral Adipose Tissue / 中国医学科学院学报

Objective To investigate the changes of regulatory T cells (Tregs) and whether Tregs can modulate the distribution of macrophage subtypes in visceral adipose tissue in the early stage of obesity.Methods After C57BL/6 mice obesity models were successfully established,metabolic parameters and numbers of Tregs and M1/M2 macrophage were measured at 4,10,and 20 weeks.The changes of metabolic parameters and adipose tissue inflammation in obesity mice after rapamycin intervention were evaluated. Results The early-stage obesity models were successfully established.Compared with normal diet mice,high fat diet mice had significantly higher epididymal adipose tissue mass and serum leptin levels(P<0.05).However,there was no statistical difference in blood glucose and insulin levels between these two groups(All P>0.05). Macrophages infiltration in adipose tissue in high fat diet mice gradually increased with time,coincident with decrease in Treg numbers. Increased numbers of Treg,improved metabolic parameters,and decreased ratio of M1/M2 can be seen after rapamycin intervention in mice.Conclusion The decrease of Tregs in the early stage of obesity may contribute to abnormal distribution of macrophage subtypes in visceral adipose.

Changes in fasting serum cortisol levels in adolescents with type 1 diabetes and elevated depressive symptoms / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes in serum cortisol levels in adolescents with type 1 diabetes (T1DM) and elevated depressive symptoms.</p><p><b>METHODS</b>Twenty-eight adolescents with T1DM and 31 healthy peers were assessed for depressive symptoms using a depression self-rating scale developed by the Epidemiological Survey Center. Selected subjects were classified into four groups: T1DM with elevated depressive symptoms group (n=15), T1DM without elevated depressive symptoms group (n=13), elevated depressive symptoms without T1DM group (n=15), and normal control group (n=16). Fasting blood samples were collected in the morning, and the levels of serum cortisol were compared among the four groups. The correlations of serum levels of cortisol and glycosylated hemoglobin A1c (HbA1c) with the score of depression self-rating scale were evaluated by Pearson correlation analysis.</p><p><b>RESULTS</b>The fasting serum cortisol levels in the 28 T1DM patients were significantly higher than in the 31 healthy peers (P<0.01). The fasting cortisol levels in the T1DM with elevated depressive symptoms group were significantly higher compared with those in the elevated depressive symptoms without T1DM group and normal control group (P<0.01). In adolescents with T1DM, serum HbA1c level was positively correlated with the score of depression self-rating scale (r=0.481, P=0.010).</p><p><b>CONCLUSIONS</b>The fasting serum cortisol levels in adolescents with T1DM and elevated depressive symptoms are significantly increased, suggesting that the patients with comorbidity of T1DM and depression develop dysfunction of the corticotropin-releasing hormone-adrenocorticotropic hormone-cortisol axis. The elevated depressive symptoms may be associated with a poor control of glucose metabolism.</p>

Current and Future Clinical Applications of Zinc Transporter-8 in Type 1 Diabetes Mellitus / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To evaluate the utility of zinc transporter-8 (ZnT8) in the improvement of type 1 diabetes mellitus (T1DM) diagnosis and prediction, and to explore whether ZnT8 is a potential therapeutic target in T1DM.</p><p><b>DATA SOURCES</b>A search was conducted within the medical database PubMed for relevant articles published from 2001 to 2015. The search terms are as follows: "ZnT8," "type 1 diabetes," "latent autoimmune diabetes in adults," "type 2 diabetes," "islet autoantibodies," "zinc supplement," "T cells," "β cell," "immune therapy." We also searched the reference lists of selected articles.</p><p><b>STUDY SELECTION</b>English-language original articles and critical reviews concerning ZnT8 and the clinical applications of islet autoantibodies in diabetes were reviewed.</p><p><b>RESULTS</b>The basic function of ZnT8 is maintaining intracellular zinc homeostasis, which modulates the process of insulin biosynthesis, storage, and secretion. Autoantibodies against ZnT8 (ZnT8A) and ZnT8-specific T cells are the reliable biomarkers for the identification, stratification, and characterization of T1DM. Additionally, the results from the animal models and clinical trials have shown that ZnT8 is a diabetogenic antigen, suggesting the possibility of ZnT8-specific immunotherapy as an alternative for T1DM therapy.</p><p><b>CONCLUSIONS</b>ZnT8 is a novel islet autoantigen with a widely potential for clinical applications in T1DM. However, before the large-scale clinical applications, there are still many problems to be solved.</p>

Effect of high-fat or high-glucose diet on obesity and visceral adipose tissue in mice / 中国医学科学院学报

<p><b>OBJECTIVE</b>To investigate the effect of high-fat or high-glucose diet on obesity and visceral adipose tissue in C57BL/6 mice.</p><p><b>METHODS</b>Four-week-old C57BL/6 mice were allocated into normal diet group,high-fat diet group,and high-glucose diet group according to the random number table until 20 weeks old. Body weight,epididymal adipose tissue weight,blood leptin,fat infiltration in liver,M1/M2 macrophage subtypes,and monocyte chemoattractant protein-1 mRNA in epididymal adipose tissues were measured.</p><p><b>RESULTS</b>Compared with normal diet group,body weight,epididymal adipose tissue weight,and leptin concentration in high fat diet group at 20 weeks were significantly increased (P < 0.05),and oil red O staining showed more prominent adipocyte infiltration in liver in high-fat diet group than those in normal diet and high-glucose diet group. However,no apparent differences were seen in high-glucose diet group at 20 weeks in terms of body weight,epididymal adipose tissue weight and leptin concentration. In high-fat diet group,the macrophages infiltration in epididymal adipose tissue increased with time and the percentage of M2 macrophage decreased in high-fat diet group than that in high-glucose diet group(P<0.05). Compared with normal diet group,monocyte chemoattractant protein-1 mRNA expression increased significantly in high-fat diet group(P<0.05). In high-glucose group,however,no significant differences were discerned (P > 0.05).</p><p><b>CONCLUSION</b>High-fat diet,rather than 60% high glucose diet,will lead to obesity and macrophage infiltration in adipose tissues.</p>

Effectiveness of different waist circumference cut-off values in predicting metabolic syndrome prevalence and risk factors in adults in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the effectiveness of waist circumference cut-off values in predicting the prevalence of metabolic syndrome (MetS) and risk factors in adults in China.</p><p><b>METHODS</b>A cross-sectional survey was condcuted in 14 provinces (autonomous region, municipality) in China. A total of 47,325 adults aged⋝20 years were selected by multistage stratified sampling, and questionnaire survey and physical and clinical examination were conducted among them. MetS was defined according to the International Diabetes Federation (IDF) criteria and modified IDF criteria.</p><p><b>RESULTS</b>The age-standardized prevalence of MetS was 24.2% (22.1% in men and 25.8% in women) and 19.5% (22.1% in men and 18.0% in women) according to the IDF criteria and modified IDF criteria respectively. The age-standardized prevalence of pre-MetS was 8.1% (8.6% in men and 7.8% in women) according to the modified IDF criteria. The prevalence of MetS was higher in urban residents than rural residents and in northern China residents than in southern China residents. The prevalence of central obesity was about 30% in both men and women according to the ethnicity-specific cut-off values of waist circumference for central obesity (90 cm for men and 85 cm for women). Multivariate regression analysis revealed no significant difference in risk factors between the two MetS definitions.</p><p><b>CONCLUSION</b>Using both the modified IDF criteria and ethnicity-specific cut-off values of waist circumference can provide more useful information about the prevalence of MetS in China. Conclusion Using both the modified IDF criteria and ethnicity-specific cut-off values of waist circumference can provide more useful information about the prevalence of MetS in China.</p>

Change of glutamic acid decarboxylase antibody and protein tyrosine phosphatase antibody in Chinese patients with acute-onset type 1 diabetes mellitus / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Glutamic acid decarboxylase antibody (GADA) and protein tyrosine phosphatase antibody (IA-2A) are two major autoantibodies, which exert important roles in the process of type 1 diabetes mellitus (T1D). Our study aimed to investigate the changes in positivity and titers of GADA and IA-2A during the course of Chinese acute-onset T1D patients and their relationships with clinical features.</p><p><b>METHODS</b>Two hundreds and forty-seven Chinese newly diagnosed acute-onset T1D patients were consecutively recruited. GADA and IA-2A were detected at the time of diagnosis, one year later, 3-5 years later after diagnosis during the follow-up; all the clinical data were recorded and analyzed as well.</p><p><b>RESULTS</b>During the course of acute-onset T1D, the majority of patients remained stable for GADA or IA-2A, however, a few patients changed from positivity to negativity and fewer patients converted from negativity to positivity. The prevalence of GADA was 56.3% at diagnosis, decreasing to 50.5% one year later, and 43.3% 3-5 years later while the corresponding prevalence of IA-2A were 32.8%, 31.0% and 23.3%, respectively. The median GADA titers were 0.0825 at diagnosis, declining to 0.0585 one year later and 0.0383 3-5 years later (P < 0.001), while the corresponding median titers were 0.0016, 0.0010, 0.0014 for IA-2A, respectively. Fasting C-peptide (FCP) and postprandial C-peptide 2 hours (PCP2h) levels of GADA or IA-2A negativity persistence patients were higher than those of positivity persistence and negativity conversion patients (P < 0.05) which indicated GADA or IA-2A negativity persistence T1D patients had a less loss of β cell function.</p><p><b>CONCLUSION</b>Our data suggest that repeated detection of GADA and IA-2A are necessary for differential diagnosis of autoimmune diabetes and the indirect prediction of the β cell function in Chinese patients.</p>

Hypoadiponectinemia predicts impaired endothelium-independent vasodilation in newly diagnosed type 2 diabetic patients: an 8-year prospective study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Adiponectin is an adipokine with insulin-sensitising and anti-atherogenic properties. The aim of this study was to investigate whether low adiponectin levels predict the impairment of endothelial function in newly diagnosed type 2 diabetic patients in an 8-year prospective study.</p><p><b>METHODS</b>In the prospective study, we enrolled 133 newly diagnosed type 2 diabetic patients without subclinical atherosclerosis and gave them intensive therapy; the mean treatment period was 8 years. Intensive treatment was a stepwise implementation of behavior modification and pharmacological therapy targeting hyperglycaemia, hypertension, dyslipidaemia and obesity. We measured baseline circulating adiponectin with an enzyme-linked immunosorbent assay, endothelium-dependent and -independent vasodilation by high-resolution vascular ultrasound. At year 8, 102 patients were reexamined for endothelium-dependent and -independent vasodilation.</p><p><b>RESULTS</b>Sex-adjusted adiponectin level was positively correlated with endothelium-independent vasodilation both at baseline (r = 0.150, P = 0.043) and at year 8 (r = 0.339, P = 0.001), whereas no association was found between adiponectin and endothelium-dependent vasodilation. In a stepwise multivariate linear regression model, adiponectin was an independent predictor for impaired endothelium-independent vasodilation at year 8 (P = 0.001).</p><p><b>CONCLUSIONS</b>Plasma adiponectin concentration was associated with endothelium-independent vasodilation and hypoadiponectinemia predicted the impairment of endothelium-independent vasodilation in newly diagnosed type 2 diabetic patients under multifactorial intervention. These data support the causative link of impairment of endothelium-independent vasodilation with hypoadiponectinemia.</p>

Gene expression changes in patients with fulminant type 1 diabetes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Fulminant type 1 diabetes (F1D) is a complex disease. Microarray analysis was used to identify gene expression changes and obtain understanding of the underlying mechanisms.</p><p><b>METHODS</b>Microarray analysis was performed on peripheral blood mononuclear cells from six F1D patients and six matched healthy subjects. Real-time polymerase chain reaction was used to verify the differentially expressed genes. NK cell activity was detected by methyl thiazoleterazolium assay.</p><p><b>RESULTS</b>Microarray analysis identified 759 genes differing in expression between F1D patients and controls at a false discovery rate of 0.05. Expression of TLR9, ELF4 and IL1RAP were verified and consistent with changes in microarray results. NK cell activity was decreased in F1D. With use of a knowledge base, differentially expressed genes could be placed within different pathways with predicted functions including interleukin-1, and tumor necrosis factor-α signaling.</p><p><b>CONCLUSIONS</b>These results identify several genes indicating possible mechanisms in F1D. NK cell dysfunction resulting from changes in expression of TLR9, ELF4 and IL1RAP, and pathways of interleukin-1 and tumor necrosis factor-α signaling might be involved in F1D through inducing β-cell dysfunction.</p>

Duloxetine versus placebo in the treatment of patients with diabetic neuropathic pain in China / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Duloxetine, a selective serotonin and noradrenaline reuptake inhibitor, has been shown to be effective in treatment of diabetic peripheral neuropathic pain and approved for the management of patients with diabetic peripheral neuropathic pain (DPNP) in the United States, European Union, and many other countries. This study assessed the efficacy and safety of duloxetine in Chinese patients with diabetic peripheral neuropathic pain.</p><p><b>METHODS</b>This double-blind, randomized, placebo-controlled, flexible-dose study treated adult patients with diabetic peripheral neuropathic pain and baseline Brief Pain Inventory (BPI) 24-hour average pain severity ratings ≥ 4 with duloxetine 60 mg to 120 mg once daily or placebo for 12 weeks. Dose adjustments of duloxetine or matching placebo were based upon investigator's judgment of clinical response. Change from baseline to endpoint in BPI average pain was the primary efficacy outcome. Secondary outcome measures included BPI-severity and -Interference, Patient Global Impression of Improvement, Clinical Global Impressions of Severity, EuroQol: 5 Dimensions, Athens Insomnia Scale, and safety measures.</p><p><b>RESULTS</b>Of 215 patients randomized, 88.4% and 82.1% of patients in placebo and duloxetine groups, respectively, completed the study. Mean change from baseline to endpoint in BPI average pain was not statistically different between the treatment groups (P = 0.124). Duloxetine- treated patients showed significantly greater pain reduction compared with those in placebo group at weeks 1, 2, and 4 (P = 0.004, P = 0.009, and P = 0.006, respectively), but not at weeks 8 (P = 0.125) and 12 (P = 0.107). Duloxetine-treated patients experienced statistically significant improvement in Patient Global Impression of Improvement, Clinical Global Impression of Severity, area under the curve for pain relief, BPI-severity pain right now, and BPI-interference walking ability. Patients treated with duloxetine 120 mg once daily showed significantly greater pain reduction on the Brief Pain Inventory average pain score relative to placebo. Duloxetine-treated patients reported nausea, somnolence, anorexia, and dysuria significantly more than placebo.</p><p><b>CONCLUSIONS</b>Although the primary study endpoint was not achieved, the overall observed response pattern suggests the efficacy of duloxetine in the treatment of Chinese patients with diabetic peripheral neuropathic pain. The safety profile for duloxetine is similar to that reported in other global trials.</p>

Relation between insulin resistance and glutamic acid decarboxylase antibody titers in latent autoimmune diabetes in adults / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relation between insulin resistance and glutamic acid decarboxylase antibody (GAD-Ab) titers in latent autoimmune diabetes in adults (LADA).</p><p><b>METHODS</b>The patients with phenotypic type 2 diabetes were screened for GAD-Ab positivity, and the 141 positive patients were divided into two subgroups according to the GAD-Ab titer, namely the high-titer group (LADA-1 subtype) and low-titer group (LADA-2 subgroup). The clinical features and insulin resistance were compared between the two groups. Insulin resistance was calculated by HOMA 2 software, and GAD-Ab and C peptide were determined with radioligand and radioimmune assay, respectively.</p><p><b>RESULTS</b>Compared with low-titer LADA patients, the patients with high titers had younger age of onset, lower BMI, higher HbA1c level, and worse fasting and postprandial C peptide levels. The insulin resistance index by HOMA 2 was significantly lower in LADA-1 group than in LADA-2 group (1.6-/+1.1 vs 2.1-/+1.1, P=0.001). The HOMA2-IR index showed a negative correlation to GAD-Ab titer.</p><p><b>CONCLUSION</b>The degree of insulin resistance is correlated to GAD-Ab titers in LADA, and low titer patients have higher insulin resistance level.</p>

Glitazones protects beta cell function from cytotoxic cytokines through PPAR gamma-dependent mechanisms / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the protective effects of glitazones on islet beta cells and PPAR gamma dependence of such effects.</p><p><b>METHODS</b>IL-1beta and IFN-gamma were used to treat NIT-1 cells, a beta cell line, to induce beta cell damage. The cells were pretreated with rosiglitazone and pioglitazone at different concentrations to study the protective effects of these drugs. The cell apoptosis rate was determined with Annexin V-FITC by flow cytometry, and the insulin secretion capacity of the cells was assessed with ELISA. GW9662 and PPARgamma-SiRNA were used to specifically inhibit PPAR to investigate the PPAR gamma-dependent mechanisms.</p><p><b>RESULTS</b>Rosiglitazone and pioglitazone at 10 micromol/L could significantly decrease the apoptosis of beta cells induced by the cytokines (apoptotic rates of 13.99% and 16.67% vs 51.33%, P<0.01). Rosiglitazone at 10 micromol/L and pioglitazone at 20 micromol/L were less effective than 10 micromol/L rosiglitazone and pioglitazone. The insulin secretion of the cytokine-treated cells decreased from 8.5-/+0.6 ng/ml of the control group to 3.6-/+0.5 ng/ml, while rosiglitazone and pioglitazone could increase the insulin secretion to 6.8-/+0.7 ng/ml and 5.9-/+0.9 ng/ml, respectively. When PPAR gamma was specifically inhibited by GW9662 and PPARgamma-SiRNA, the protective effects of rosiglitazone and pioglitazone were almost undetectable, and the apoptotic rate increased and insulin secretion decreased to the level of the cytokine-treated cells.</p><p><b>CONCLUSION</b>Glitazones can protect beta cells from apoptosis and impairment of insulin secretion function resulting from the cytotoxic cytokines via a PPAR gamma-dependent mechanism.</p>

Micro-plate radiobinding assay of autoantibody to glutamic acid decarboxylase / 中华核医学杂志

Objective The purpose of this study was to develop a high-throughput micro-plate radiobinding assay (RBA) of glutamic acid decarboxylase antibody (GAD-Ab) and to evaluate its clinical application. Methods 35labeled GAD65 antigen was incubated with sera for 24 h on a 96-well plate, and then transferred to the Millipore plate coated with protein A, which was washed with 4℃ PBS buffer, and then counted by a liquid scintillation counter. The GAD-Ab results were expressed by WHO standard unit (U/ml). A total of 224 healthy controls, 162 patients with type 1 diabetes mellitus(T1DM) and 210 patients with newly diagnosed type 2 diabetes (T2DM) were recruited. A total of 119 TI DM and healthy cases with gradually changing GAD-Ab levels were selected to compare the consistency of micro-plate RBA with conventional radioligand assay (RLA). Blood samples were obtained from the peripheral vein and finger tip in 32 healthy controls, 35 T1DM and 24 T2DM patients, and tested with micro-plate RBA and then compared with the conventional RLA to investigate the reliability of finger tip sampling. Linear correlation,student's t-test, variance analysis and receiver operating characteristic (ROC) curve were performed using SPSS 11.5. Results (1) The optimized conditions of micro-plate RBA included 2 μl serum incubated with3 ×104 counts/min 35S-GAD for 24 h under slow vibration, antigen-antibody compounds washed 10 times by 4℃ PBS buffer, and radioactivity counted with Optiphase Supermix scintillation liquid. (2)The intra-batch CV of the micro-plate RBA was 3.8%- 10.2%, and the inter-batch CV was 5.6%- 11.9%. The linearity analysis showed a good correlation when the GAD-Ab in serum samples ranged from 40.3 to 664 U/ml and the detection limit of measurement was 3.6 U/ml. The results from Diabetes Autoantibody Standardization Program (DASP) 2005 showed that the sensitivity and specificity for GAD-Ab were 78% (39 positive among 50 new-onset T1DM) and 98% (2 positive among 100 healthy controls). The results of GAD-Ab obtained with micro-plate RBA and RLA were closely correlated (r=0.915,P<0.001) with a high concordance level of 97.5% and a Kappa value of 0.95. (3)TI DM and T2DM patients showed higher positive rates for GAD-Ab than the healthy controls(46.9% and 5.2% vs 0.89% ,X2=123.5 and 10. 1 ,P <0.001 and <0.01, respectively). (4)The consistency of GAD-Ab measurement with RBA using finger tip blood and RLA measurement using venous blood was 96.7% (r =0.946,P <0.001, Kappa value: 0.905). Conclusions The micro-plate RBA of GAD-Ab has high sensitivity, specificity and reproducibility, and can be measured with finger tip blood sampling. It might be a better alternative for clinical practice.

Multicenter clinical study on the efficacy and safety of inhalable insulin aerosol in the treatment of type 2 diabetes / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A new inhalable insulin aerosol (Inh-Ins) was developed in China. The aim of this multicenter clinical study was to evaluate the efficacy and safety of this new Inh-Ins as a treatment of type 2 diabetes. Regular porcine insulin (RI) was used as a control.</p><p><b>METHODS</b>This study is a prospective, randomized, open-label, parallel-group multicenter clinical trial in which 253 qualified patients with type 2 diabetes received the insulin Glargine daily at bedtime plus either a pre-meal Inh-Ins or a pre-meal subcutaneous RI for 12 weeks. HbA1c, fasting plasma glucose (FPG), the 1-hour-postprandial blood glucose (1hPBG) and the 2-hour-postprandial blood glucose (2hPBG) were measured. Events were monitored for adverse effects.</p><p><b>RESULTS</b>After 12 weeks, the HbA1c decreased significantly from baseline in both treatment groups, with no significant difference between the two regimens. In the Inh-Ins group, FPG, both 1hPBG and 2hPBG significantly declined from baseline after the 8th- and 12th-weeks of treatment. The reduced values of FPG or 1hPBG between the two groups showed a more significant hypoglycemic effect with the Inh-Ins than the RI. After 12 weeks, the pulmonary carbon monoxide diffusing capacity (DLco) was significantly lower in Inh-Ins group than in the RI. The main side effects of Inh-Ins were coughing, excessive sputum, and hypoglycemia.</p><p><b>CONCLUSIONS</b>Inh-Ins was effective in decreasing HbA1c like the RI. It was better in lowering the FPG and the 1hPBG than the RI. Its main side effects were coughing, excessive sputum, and hypoglycemia. Also, Inh-Ins slightly impaired DLco.</p>

Construction and identification of 2 secreted human GAD65 fragment DNA vaccines / 中南大学学报(医学版)

OBJECTIVE@#To construct and identify 2 secreted human GAD65 fragment DNA vaccines.@*METHODS@#The GAD(190-315), GAD(490-570) cDNA and hIL-2 signal peptide cDNA were linked through overlapping PCR, respectively. The fusion gene was cloned into eukaryotic expression vector pBudCE4.1. After the DNA vaccine being determined to contain the correct target nucleotide sequence, the expression of fusion proteins was detected by Western blot.@*RESULTS@#The nucleotide sequence of the cloned gene was the same as the reported sequence, and their open reading fragment was correct. The products of these DNA vaccines were expressed and secreted in eukaryotic cell using Western blot.@*CONCLUSION@#The pBudCE4.1/SGAD(190-315) and pBudCE4.1/SGAD(490-570) secreted human GAD65 fragment DNA vaccines were successfully constructed, which is a foundation for immune prevention of type 1 diabetes.

Oral administration of insulin inhibits islet beta cell apoptosis and prevents diabetes in NOD mice / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of oral administration of insulin on insulitis beta cell apoptosis and diabetes in non-obese diabetic (NOD) mice, and to explore the mechanism of immune tolerance induced by insulin.@*METHODS@#Eighty-six female NOD mice were randomly divided into an insulin group (n=43) and a phosphate buffered saline (PBS) group (n=43). From 4 weeks of age, the recombinant human insulin (Humulin R) 1 mg (70 microL) was administrated in the oral insulin group and 70 microL PBS in the control group respectively, twice per week before 12 weeks of age and then once weekly until 30 weeks. Insulitis and beta cell apoptosis of islets were observed at 12 weeks. IL-4 and IFN-gamma in the sera were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of I-Abeta(g7), IL-4, IFN-gamma, IL-1beta, Fas and TGF-beta mRNA of islets, and IL-4, IFN-gamma, TGF-beta mRNA of Peyer's patch were measured by reverse transcription-polymerase chain reaction (RT-PCR) at 12 weeks.@*RESULTS@#The incidences in the insulin group were significantly lower than those in the PBS group (55.6% vs 85.7% at 30 weeks, 70.4% vs 96.4% at 52 weeks, P<0.05). The insulitis scores in the insulin group were lower than those in the PBS group, but there was no statistical significance. Fas expression on islets and apoptotic beta cell rates in the insulin group were lower than those in the PBS group (P<0.05). In the insulin group, serum IL-4 levels were higher, and IFN-gamma levels were lower than those in the PBS group (P<0.05). The levels of I-Abeta(g7), IFN-gamma, IL-1beta and Fas mRNA transcription in islets and IFN-gamma mRNA transcription in Peyer's patch were both lower in the insulin group, and IL-4, TGF-beta mRNA levels were higher than those in the PBS group (P<0.05).@*CONCLUSION@#The specific autoantigen insulin may induce the immune tolerance and prevent the diabetes in NOD mice, but it cannot block the progression of insulitis. Oral administration of insulin can induce the regulatory T cells, and make Th1 to Th2 cytokine shifts in the system and islets, thus preventing the Fas-mediated beta-cell apoptosis and diabetes.

Inhibition of islet beta cell apoptosis and prevention diabetes by subcutaneous administration of insulin in NOD mice / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effects of subcutaneous administration of insulin on insulitis,beta cell apoptosis and diabetes in non-obese diabetic (NOD) mice, and to explore the mechanism of immune tolerance induced by insulin.@*METHODS@#Sixty female NOD mice were randomly divided into insulin group (n=32) and phosphate buffered saline (PBS) group (PBS group, n=28). Insulin was subcutaneously injected with humulin N (60 microL, 6U)+IFA (60 microL) at 4, 12, 20, and 28 weeks respectively, while the PBS group received PBS (60 microL) + IFA (60 microL). Insulitis and beta cell apoptosis of islets were observed at 12 weeks. IL-4 and IFN-gamma in the sera were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of I-Abeta(g7), IL-4, IFN-gamma, IL-1beta, and Fas mRNA of islets were measured by reverse transcription-polymerase chain reaction (RT-PCR) at 12 weeks.@*RESULTS@#The incidences in the insulin group were significantly lower than those in the PBS group (21.4% vs 71.4% at 30 weeks, 28.6% vs 85.7% at 52 weeks, P<0.05). The insulitis scores in the insulin group were lower than those in the PBS group, but there was no statistical significance. Fas expression on islets and apoptotic beta cell rates in the insulin group were lower than those in the PBS group (P<0.05). In the insulin group, serum IL-4 levels were higher, but IFN-gamma levels were lower than those in the PBS group (P<0.05). The levels of I-Abeta g7, IFN-gamma, IL-1beta and Fas mRNA transcription in islets were lower in insulin group, but IL-4 mRNA levels were higher than those in the PBS group (P<0.05).@*CONCLUSION@#The specific autoantigen insulin may induce immune tolerance and prevent diabetes in NOD mice, but it can't block the progression of insulitis. Subcutaneous administration of insulin can induce the regulatory T cells, and make Th1 to Th2 cytokine shifts in system and islets, thus preventing the Fas-mediated beta-cell apoptosis and diabetes.

Cross-sectional study of the relation between carboxypeptidase-H antibody and islet beta cell function in patients with latent autoimmune diabetes in adults / 中南大学学报(医学版)

OBJECTIVE@#To explore the relation between carboxypeptidase-H antibody (CPH-Ab) and islet beta cell function in patients with latent autoimmune diabetes in adults (LADA) and to further confirm the diagnostic value of CPH-Ab for LADA.@*METHODS@#Five hundred and forty-five patients who were initially diagnosed as Type 2 diabetes mellitus (T2DM) were tested with CPH-Ab and GAD-Ab by radioligand assay (RLA). T2DM patients, according to CPH-Ab and GAD-Ab status, were divided into CPH-Ab(+) group, GAD-Ab(+) group, and Ab(-) group to compare their islet beta cell function [represented by fasting C-peptide (FCP) and 2h postprandial C-peptide (2hCP)]. The relation between CPH-Ab and islet beta cell function in LADA was analyzed.@*RESULTS@#The fasting C-peptide level in CPH-Ab(+) patients was between that of GAD-Ab(+) patients and that of Ab(-) patients (P0.05). Corrected by concomitant variables including age, age at onset, duration of disease, and sex, the differences among the 3 groups were statistically significant (both P0.05).@*CONCLUSION@#The effect of CPH-Ab is less marked than that of GAD-Ab on islet beta-cell function in LADA patients. The value of CPH-Ab for the failure of islet beta-cell function in LADA should be determined prospectively.

Effect of complete Freund's adjuvant on islet beta cell apoptosis and its mechanism in non-obese diabetic mice / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of complete Freund's adjuvant (CFA) on islet beta cell apoptosis in preventing diabetes in non-obese diabetic (NOD) mice, and the influence on apoptotic related-gene expression.@*METHODS@#Four-week-old female NOD mice were randomly divided into Group CFA (n=5) and Group saline (NS) control (n=5). Mice in Group CFA were injected in the hind footpad with 50 microL CFA and mice in Group NS with 50 microL NS. Blood sugar was monitored and diabetes was diagnosed if blood sugar was higher than 11.1 mmol/L for 2 continuous days in the NOD mice. The mice were sacrificed when diagnosed as diabetes or at 30 weeks of age. Pancreatic sections were made for: evaluation of insulitis severity with HE staining; counting of apoptotic beta cells with the terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labeling (TUNEL) method, and ABC immunohistochemical double labeling; counting of Fas, FasL and Bcl-x positive cells respectively with ABC immunohistochemical method.@*RESULTS@#By 30 weeks of age, none of the 5 CFA-treated mice, compared with 3 of the 5 control mice, had developed diabetes. The insulitis score was lower (1.820+/-0.962 vs. 3.020+/-1.040, P<0.05), the rates of apoptotic beta cells, Fas positive cells and FasL positive cells were lower [(10.2+/-2.8)% vs. (15.9+/-6.5)%, (54.9+/-14.5)% vs. (75.7+/-12.9)%, (20.3+/-10.4)% vs. (27.9+/-12.0)%, P<0.05), and the Bcl-x positive cell rate was higher [(74.9+/-10.7)% vs. (66.0+/-18.3)%, P<0.05] in the CFA-treated group than those in the NS-treated group respectively.@*CONCLUSION@#CFA may inhibit beta cell apoptosis in NOD mice by regulating Fas, FasL and Bcl-x expression on cells within islets.

Risk factors for the progress of subclinical atherosclerosis in newly diagnosed type 2 diabetics with multifactorial intervention / 中华全科医师杂志

Objective To investigate risk factors for the progress of subclinical atherosclerosis (AS)in newly diagnosed type 2 diabetics with muhifactorial intervention.Methods One hundred and fifty- six patients of type 2 diabetes,aged 35～70 years,with course of less than one year and without subclinical AS,were observed prospectively.After two-year intervention based on anti-platelet therapy integrated with intensive control of blood glucose,blood lipid,blood pressure and body weight,dynamic changes in all metabolic indicators and subclinical AS in the patients and differences between those with subclinical AS and without it were analyzed to study the risk factors for its progress by logistic regression analysis.Results There were no significant differences in intima-medial thickness(IMT)of common carotid artery(CCA) and femoral artery(FA)in the patients between baseline and two years after intervention,but those in them were significantly increased two years after intervention than those one year after intervention(P＜0.O1). Two years after intervention,increased IMT or atherosclerotic plaques could be found in 45 of 156 patients (28.8%),significantly higher than those one year after intervention(11.5%,P＜0.01).Levels of glycosylated hemoglobin Alc(HbAlc),total cholersterol(TC),high-density lipoprotein-cholesterol (HDL-C)and HOMA-insulin resistance(IR)showed an increased trend two years after intervention,as compared with those one year after intervention(P＜0.01).Proportions of those with normal level of HbAlc two years after intervention was significantly lower than that one year after intervention(P＜0.01). Proportions of those with normal level of HbAlc and low-density lipoprotein-cholesterol(LDL-C)were significantly lower in patients with subclinical AS than those without it two years after intervention(P＜0.01).Logistic regression analysis showed that relative risk(RR)for subclinical AS could reduce by 83% with normal level of LDL-C and 59% by normal HbAlc,respectively,but there was an 82% increase in RR for it with an increase of age by ten years.Conclusions Subclinical AS could not be absolutely prevented in patients with newly diagnosed type 2 diabetics after two-year intensive multifactorial intervention.Subclinical AS could present a progressive trend with time,as well as levels of blood glucose and blood lipid.Levels of LDL-C and HbAlc,as well as age,were major risk factors for occurrence of subclinical AS in patients with newly-diagnosed type 2 diabetes.

Study on the postive frequency and distribution of glutamic acid decarboxylase antibody in phenotypic type 2 diabetec patients / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the positive frequency and distribution of glutamic acid decarboxylase antibody(GAD-Ab) in phenotypic type 2 diabetic(T2DM) patients.</p><p><b>METHODS</b>Sera of 2035 phenotypic T2DM patients were screened for GAD-Ab with radioligand assay. The positive frequency of GAD-Ab and its relation with clinical features were analyzed.</p><p><b>RESULTS</b>(1) The positivity of GAD-Ab in clinic-based, phenotypic T2DM patients was 7.1% (145/2035), comparable to that of data from Caucasians as shown by UKPDS(8.7% vs. 9.8%, P = 0.391) and ADOPT (8.0% vs. 4.2%, P = 0.000) but higher than that of Japanese in Ehime study(7.1% vs. 3.8%, P = 0.000). (2) The positive frequency and distribution of GAD-Ab titer were related to clinical features, including age at onset, body mass index (BMI) and fasting C peptide levels. Patients with younger age at onset (0.33 vs. 0.11, P < 0.05), less BMI (0.34 vs. 0.10, P < 0.05) and lower C peptide levels (0.38 vs. 0.11, P < 0.05) would have higher GAD-Ab titers.</p><p><b>CONCLUSION</b>(1)The positivity of GAD-Ab in adult-onset phenotypic T2DM in Chinese was similar to that of Caucasians but higher than that of the Japanese. (2) The distribution of GAD-Ab titers was associated with clinical features, with high GAD-Ab titers for those having younger age at onset, less BMI and lower C peptide levels.</p>