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Objective To investigate the mechanism of pioglitazone preventing diabetes and the role of nuclear factor of actived T cells (NFAT) on non-obese diabetic(NOD) mice .Methods (1)Female NOD mice at 4 weeks of age were randomly divided into pioglitazone group(n=21) and control group(n=21) .The accumulative diabetes incidence was followed-up to 30 weeks of age in each group of NOD mice .(2)Pancreas were removed from NOD mice at 12 weeks of age in each group(n=15) to score insulitis se-verity by routine HE staining .IL-4 ,IFN-γand peroxisome proliferator-activated receptor γ(PPARγ) mRNA levels in spleens were tested by RT-PCR .IL-4 and IFN-γlevels in sera ,the activity of PPARγand NFATc1 nuclear protein in spleens were measured by enzyme linked immunosorbent assay (ELISA) .Results (1) At 15 weeks of age ,the diabetes incidence was 4 .76% in pioglitazone group ,and 33 .33% in control group(P0 .05) .(2) At 12 weeks of age ,the insulitis score in pioglitazone group was lower than that in control group[(1 .79 ± 0 .75) vs .(2 .38 ± 0 .66) ,P<0 .05] .(3) IFN-γ mRNA level in pioglitazone group was lower than that in control group[(0 .16 ± 0 .07) vs .(0 .53 ± 0 .26) ,P<0 .05] ,and PPARγmRNA level in pioglitazone group was higher than that in control group(0 .91 vs .0 .25 ,P<0 .05) .(4)IFN-γ level in pioglitazone group was lower than that in control group [(561 .05 ± 78 .61)pg/mL vs .(666 .43 ± 28 .42)pg/mL ,P<0 .05] .(5)At 12 weeks of age ,the spleen PPARγnuclear protein activity in pioglitazone group was higher than that in control group [(0 .05 ± 0 .01) vs .(0 .02 ± 0 .01) ,P<0 .05)] ,and NFATc1 nuclear protein activity was low-er than that in control group[(0 .23 ± 0 .04) vs .(0 .33 ± 0 .04) ,P<0 .05] .Conclusion Pioglitazone could activate PPARγ nuclear protein ,inhibit activity of NFATc1 nuclear protein ,downregulate IFN-γ,diminish Th cells deviating to Th1 ,and sequently prevents insulitis and diabetes onset in NOD mice .
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Este estudo teve por objetivo propor um protocolo para o manejo de resíduos de serviços de saúde gerados em domicílios de indivíduos com diabetes Mellitus usuários de insulina, com foco nos resíduos biológicos, químicos e perfurocortantes. Constitui-se de uma pesquisa descritivo-exploratória realizada em um núcleo de Saúde da Família de Ribeirão Preto - SP. Participaram do estudo 26 usuários de insulina. Os dados foram coletados por meio do SIAB e entrevistas com os sujeitos selecionados, durante o mês de julho de 2010. Os dados foram analisados por meio da estatística descritiva. Os resultados revelaram um descarte inadequado dos resíduos de serviços de saúde gerados em domicílios de usuários de insulina. Essa situação requer a adoção de protocolos com critérios para o controle da distribuição e descarte de seringas, favorecendo a minimização da geração de resíduos de serviços de saúde em domicílios. O uso racional de seringas também pode contribuir para a aquisição de materiais em quantidade adequada pelos usuários de insulina. Os conhecimentos gerados nesta pesquisa, relacionados ao manejo de resíduos de serviços de saúde gerados em domicílios de usuários deinsulina, pode despertar a implementação de ações de educação em saúde e embasar o desenvolvimento de protocolos nas unidades de Saúde.
This study aimed to propose a protocol to the management of medical waste generated in households of individuals with diabetes mellitus who are insulin users, focusing on biological, chemical and sharp waste. This is a descriptive and exploratory study carried out in a Family Health Center in the city of Ribeirão Preto, State of São Paulo, Brazil. The sample was composed of 26 users of insulin. Data were collected through the Information System of Primary Care and interviews with selected participants during the month of July 2010. Data were analyzed using descriptive statistics. Results showed the improper disposal of medical waste generated in households of insulin users. This situation requires the adoption to protocols for the control of distribution and disposal of syringes, which could help to minimize the generation of medical waste in households. The rational use of syringes may avoid the wasting and it is a way to ensure the acquisition of adequate amounts of material by the insulin users. The knowledge generated in this research related to the problem of the management of medical waste generated in households of insulin users may to contribute to the development of actions in health education and protocols in health units.
Este estudio tuvo el objetivo de proponer un protocolo para el manejo de residuos de servicios de saludgenerados en domicilios de individuos con diabetes mellitus, usuarios de insulina, centrándose en los residuos biológicos, químicos y corto-punzantes. Se trata de una investigación descriptiva-exploratoria, realizada en un Núcleo de Salud de la Familia en la ciudad de Ribeirão Preto, estado de São Paulo, Brasil. Participaron delestudio 26 usuarios de insulina. Los datos fueron recogidos por medio del Sistema de Información de Atención Primaria y entrevistas con los sujetos seleccionados, durante el mes de julio de 2010 y analizados utilizando la estadística descriptiva. Los resultados revelaron la eliminación inadecuada de los residuos de servicios de salud generados en domicilios de usuarios de insulina. Esta situación requiere la adopción de protocolos con criterios para el control de la distribución y eliminación de jeringas, favoreciendo la disminución de la generación de residuos de servicios de salud en domicilios. El uso racional de jeringas también puede contribuir para la adquisición de materiales en cantidad adecuada por los usuarios de insulina. Los conocimientos generados en esta investigación, relacionados al manejo de residuos de servicios de salud generados en domicilios de usuarios de insulina, puede contribuir para la implementación de acciones de educación en salud y basar el desarrollo de protocolos en las Unidades de Salud.
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Objective Insulin autoantibody (IAA) is known to exist in sera of type 1 diabetes mellitus (T1DM) patients and pre-T1DM individuals. The aim of this study was to establish a novel microtiter plate radioimmunoassay (RIA) for IAA and evaluate its clinical value. Methods Diluted 125Ⅰ-insulin was mixed with 5 ul serum samples in a 96-well microtiter plate and then incubated for 72 h on an orbital plate shaker (4℃). The immunocomplexes were transferred to another protein a coated Millipore plate, and then the plate was washed with Tri-Buffered Saline Tween-20 (TBT) buffer. Counts per minute (CPM) was measured with liquid scintillation and luminescence counter. The positive cut-off point of IAA index was defined as ≥0.06 based on the 99-percentile of the distribution in 317 healthy individuals. The specificity and sensitivity of the assay were calculated from the samples provided by the fourth Diabetes Autoantibodies Standardization Program (DASP 2005). The IAA levels were determined in 71 T1 DM and 551 newly diagnosed type 2 diabetes (T2DM) patients, and 317 healthy controls. The t test, non-parametric test, x2 test and linear correlation analysis were performed on the data using SPSS 11.5 software. The concordance rate was estimated with Kappa value. Results (1) The optimized testing condition was described as 2×104 CPM of 125Ⅰ-insulin, 5 ul serum sample and slowly horizontal shaking for 72 h. (2) The intra-assay CV was 4.8%-8.9% and inter-assay CV was 6.4%-10.5%. Based on DASP 2005 samples, the specificity and sensitivity of the assay were 97% (97/100) and 50% (25/50), respectively. Ninety-six serum samples with different IAA levels were selected and tested to compare between our new method and a domestic IAA RIA kit. The results showed that the IAA indices from the two methods were positively correlated (r= 0.678, P<0.001). The concordance rate was 72.9 %(Kappa value=0.402). There were 25 samples with discordant results, which were positive for IAA titer using the corresponding microtiter plate RIA but negative using the novel RIA kit. (3) In TIDM group the positive rate of IAA was 19.7% (16/71), higher than the healthy controls (0.9%, x2=54.36, P<0.001). The subgroup of T1DM children (with 0-9 years) showed the highest IAA positive rate (55.6% ,x2=4.85, P<0.05). In T2DM group the frequency of IAA was 1.5% (8/551), which had no significant difference comparing with that of healthy controls (x2= 0.95, P >0.05). Conclusions Our proposed microtiter plate RIA method for IAA is highly sensitive and specific, likely to be feasible for clinical application. The frequency of IAA is high in children with T1DM.
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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.
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O artigo tem como objetivo principal caracterizar os perfis glicêmicos domiciliares de pacientes com diabetes mellitus tipo1, a partir de um esquema de monitorização proposto, e adotá-los como estratégia de ajuste nas doses de insulina. Foram realizados 3259 testes, 781 antes do café, 752 antes do almoço, 765 antes do jantar, 740 antes de deitar e 221 pela madrugada. A média das glicemias nestes períodos ultrapassaram os limites superiores satisfatórios em 6,87%, 3,83%, 11,37%, 30,50% e 19,28% respectivamente. Estes dados forneceram subsídios para ajustes nos esquemas insulinoterápicos. Os níveis HbA1c não mudaram de forma significante com os ajustes realizados porém, foram mantidos em 10%.
The goal of this paper is to characterize the glycemic profiles of patients with type 1 diabetes mellitus like a strategy in insulin adjustments. A total of 3259 tests were realized being 781 before breakfast, 752 before lunch, 765 before dinner and 740 before bed and 221 in the dawn. The average of the blood glucose tests in these periods oversteped the superior limits in 6,87%, 3,83%, 11,37%, 30,50% e 19,28% respectively. These data gave the conditons to make the insulin adjustments. The HbA1c levels evidenced that there was no significant statistical difference in the metabolic control, but they remained in 10%.
El objetivo principal de este artículo consiste en caracterizar los perfiles glicémicos domiciliares de los pacientes con diabetes mellitus tipo 1, a partir de un esquema propuesto de monitorización, para aplicarlo como estrategia, en los ajustes de las dosis de insulina. Fueron realizados 3.259 tests, de los cuales 781se hicieron antes del desayuno, 752 se realizaron antes del almuerzo, 765 antes de la merienda, 740 se hicieron antes de dormir y los 221 se hicieron en la madarugada. La media de las glicemias en estos períodos ultrapasaron los límites superiores satisfactorios em 6,8%, 3,83%, 11,37%, 30,50% y 19,28% respectivamente. Estos datos ofrecieron subsidios para poder ajustar los esquemas insulinoterápicos. Los niveles de HbA1c no variaron de forma significativa com los ajustes realizados y se mantuvieron em 10%.
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Adolescente , Adulto , Niño , Preescolar , Femenino , Humanos , Masculino , Glucemia/análisis , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Hipoglucemiantes/administración & dosificación , Insulina/administración & dosificación , Automonitorización de la Glucosa Sanguínea/métodos , Diabetes Mellitus Tipo 1/sangre , Monitoreo Ambulatorio/métodos , Factores de TiempoRESUMEN
Objective To probe the indication, technique and effect of combined liver pancreas transplantation. Methods Donor organs were harvested by the technique of total abdominal evisceration. Simultaneous orthotopic liver and heterotopic pancreas duodenum transplantation was performed in one patient diagnosed as having chronic hepatitis B, hepatocirrhosis, hepatic cellular cancer, and insulin dependent diabetes. Liver and pancreas graft function was monitored after transplantation. Results The recipient recovered with excellent allograft function, and insulin independence was achieved without any surgical complication. Conclusion End stage liver disease with concomitant insulin dependent diabetes is the indication for combined liver pancreas transplantation.
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Objective To evaluate clinical effect of simultaneous kidney-pancreatic transplantation(SKPT) with portal venous and enteric drainage. Method Between June 2001 and June 2004, six insulin-dependent diabetes mellitus(IDDM) and renal failure patients underwent this procedure. The venous drainage of the graft was established between donor′s portal vein and recipient′s superior mesenteric vein. The exocrine secretion was drained into proximal jejunum via side-to-side anastomosis between donor′s duodenum and recipient′s proximal jejunum. Steroids, mycophenolate mofetil, tacrolimus and Zenapax were used as immunosuppressants. Results Procedures were successful in all 6 cases. Excellent renal function and euglycemia were achieved in 4 cases. Follow-up of 4-34 months on the 4 survivers found excellent kidney-pancreatic function without any rejection episode. Two patients died perioperatively due to sepsis secondary to pancreatic leakage and drug toxicosis of excessive FK506. Conclusion Our preliminary experience suggests that simultaneous pancreas-kidney transplantation with enteric and portal drainage is reliable procedure for the treatment of IDDM with renal failure.
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Objective To construct insulin B chain DNA vaccine and to perform a vaccinating trial for preventing mice from autoimmune diabetes induced by multiple low dose streptozotocin. Methods RT PCR was used to clone insulin B chain, and insulin B chain DNA vaccine was constructed. DNA vaccine was injected into tibilias anterior muscle of C57BL/6 mice in treatment group (T), prophylactic group (P). The morphology of mice islets was investigated by pathology, blood samples were taken to determine glucose and insulin. Group T and group P were compared with group of normal control mice (C) and group of untreated diabetic mice (D). Results (1) Insulin B chain gene was cloned by RT PCR, which ligated with pcDNA3 plasmid. Insulin B chain DNA vaccine was constructed. (2) In group P, after injection of DNA vaccines, 3 (3/10) and 4 (4/10) mice became diabetic in the second week and the third, fourth week respectively, the incidence was significantly less than those of groups D and T. The insulin level showed no difference between group C and the non diabetic mice of group P. The change of pancreas of diabetic mice in group P was similar to group D. Conclusion Insulin B chain DNA vaccine can prevent pre autoimmune diabetes.
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Objective To study the effect of phosphodiesterase inhibitor pentoxifylline (PTX) on type 1 diabetes in NOD mice and its possible mechanism.Methods Blood glucose, urinary glucose, insulitis and incidence of diabetes were investigated in NOD mice treated with PTX, insulitis was observed by HE staining and the expressions of IFN ?, TNF ?, IL 10 in pancreas were measured by RT PCR technique. Results The incidence of diabetes in the PTX group was 30.0% which was significantly lower than 67.9% in the control group (P
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Objective To evaluate the significance of combined determination of glutamic acid decarboxylase antibodies(GADA),islet cell antibodies(ICA) and insulin autoantibodies (IAA) in predictive of type 1 diabetes mellitus Methods GADA, ICA and IAA were determined by enzyme linked immunosorbent assay and C peptide by radioimmunoassay Diagnose efficiency were calculated by matrix decision method Results The positive rate of GADA,ICA and IAA(68 8%,43 0% and 39 8%) were significantly elevated in the patients with type 1 diabetes mellitus compared with type 2 diabetes mellitus or normal subjects ( P
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Objective To evaluate the effect and result of xenotransplantation with alginic polylysine alginic(APA) microencapsulated neonatal swine islets for the treatment of IDDM patients.[WT5”HZ] Method [WT5”BZ] The neonatal pig islets were microencapsulated with APA technique and cultured in vitro. The secretion of insulin, glucose stimulated insulin release test and histological examination between the microencapsulated and unmicroencapsulated neonatal pig islets were compared. 3 patients with IDDM received a xenotransplantation of microencapsulated neonatal pig islets by laparoscopic procedure. The change in blood glucose level, C peptide and the dose of insulin used were observed before and after the xenotransplantation. [WT5”HZ]Results [WT5”BZ] No significant difference was found between microencapsulated and unmicroencapsulated neonatal pig islets in vitro in terms of biological activity of the islets. The levels of serum C peptide in two of the 3 recipients increased by 11 and 23 times respectively, the postoperative dose of insulin needed decreased by 63% in one, insulin independency was achieved in the other recipient.In these two cases,the microencapsulated islets have functioned effectively for up to 80 ds.[WT5”HZ] Conclusion [WT5”BZ] APA microencapsulated neonatal pig islets have good biological activity and survived while transplanted into IDDM recipients. Omentum minus cavity xenotransplantation of microencapsulated neonatal pig islets by laparoscopic technique is safe, capable of repeated transplantation.
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AIM: To investigate the intervention effect of pentoxifylline(PTX) on type 1 diabetes mellitus in non-obese diabetic(NOD)mice and explore its possible mechanism.METHODS: Eight-week-old NOD mice were treated with PTX to investigate the incidence of cyclophosphamide accelerating diabetes.The apoptosis of beta-cells was detected by TUNEL,the expressions of caspase-3 in islet of the NOD mice was checked by immunohistochemistry and the expressions of caspase-8 was determined by RT-PCR.RESULTS: The incidence of diabetes in PTX group was 40.63%,which was obviously lower than 69.70% in the control group(P
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Objective To investigate HLA-DPB1, DQB1 gene alleles and their correlation with autoantibodies in type 1 diabetes mellitus. Methods Sequence-based typing was used to determine the alleles of HLA-DPB1 and -DQB1 in 68 patients with type 1 diabetes and 50 healthy controls. Autoantibodies 〔glutamic acid decarboxylase antibody (GADA), islet cell antibody (ICA), insulin autoantibody (IAA)〕were detected by ELISA. Results Compared with the controls, the frequencies of DPB1*0402 and DQB1*0301 were significantly lower in type 1 diabetics (11.76%, 5.15% vs 30.00%, 22,00% respectively, P
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Objective To observe the clinical effects of combined islet cell and kidney transplantation in the treatment of insulin-dependent diabetes mellitus associated with end-stage renal failure.Methods Four patients with type I diabetes and ESRD received combined renal and islet cells transplantation.Clinical and metabolic data were studied during the follow-up.Results The cultured human adults islet cells were infused into the portal vein system of the 4 patients.Immunosuppression included CsA.azathioprine and prednisone.Metabolic follow-up comprised assessment of daily fasting and non-fasting blood glucose,basal C-peptide secretion,HbAIc,renal function and blood cell counts.Islet isolation yielded 25 000~48 000 equivalents(single or multiple donors).No adverse effects were seen subsequent to islet transplantation.Basal C-peptide secretion maintained at normal levels.and blood glucose and HbAIc levels were normalized throughout the observation period.The dosage of insulin were decreased by over 25% in all 4 patients after transplantation.Conclusions Combined adult islet cells and renal transplantation has a good effect in the treatment of the patients with type Ⅰ diabetes and ESRD and can be used as an effective way for treating ESRD secondary to the type Ⅰ diabetes.Postoperative efficacy is related not only to the quantlty and quality of the islet cells,but also to the rejection of the grafts.
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Objective To investigate the preventive effect of 1,25-dihydroxyvitamin D_3 〔1,25-(OH)_2D_3〕 on type 1 diabetes in cyclophosphamide-treated NOD mice. Methods Twenty female NOD mice were administered 280 mg/kg cyclophosphamide by a single intraperitoneal injection at the beginning of the experiment and then divided into two equal groups. Group 1 received intraperitoneal injection of 5 ?g/kg 1,25-(OH)_2D_3 every other day; Group 2 received the intraperitoneal injection of peanut oil in the same volume as control. When the experiment was finished on the 30th day, the incidence of diabetes and the degree of insulitis were observed. The expressions of bcl-2 and bax in islets were detected with immunohistochemical technique. The apoptosis rate of spleen T lymphocyte was also measured by flow cytometry quantitative analysis. Results Intraperitoneal injections of 1,25-(OH)_2D_3 to NOD mice reduced the incidence of diabetes (10% vs 70%, P
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Apoptosis, also known as programmed cell death,is a normal process contributing to tissue tumor during development and in the adult.During development apoptosis is involved in tissue homeostasis and eliminating nonfunctional,superfluous or harmful cells.It is an active process controlled by some gene and factors such as caspases, bcl-2, Fas/FasL and NO.Type 1 diabetes mellitus is regarded as a chronic autoimmune disease resulting from progressive destruction of insulin-producing ?-cells in the pancreas.The relationship between apoptosis with its modulation and type 1 diabetes mellitus was reviwed.
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Objective To assess the effect of fetal islet transplantation for the treatment of Type Ⅰ diabetes. Methods The pancreatic islets from human aborted embryos were cultured and implanted into the greater omentum and omental bursa of 26 patients with type Ⅰ diabetes. The function of transplanted islets was evaluated. Results After transplantation, the exogenous insulin requirement significantly decreased (P
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Expression of HLA class Ⅱ antigens(HLA-DR, DQ and DP), interleukin2 receptors(IL-2R) and transferrin receptors(TfR) of blood monocytes from 10 patients with insulin-dependent diabetes meIlitus (IDDM) were assayed with the indirect immune fluorescence technique using corresponding monoclonal antibodies and the FITC-labelled second antibody. The results showed that the number of HLA-DQ~+ monocytes was much more in diabetics than in normal controls. The percentages of HLA-DR~+ and HLA-DP~+ monocytes in diabetics were not different significantly from those in normal controls. Besides, IL-2R~+ and TfR~+ monocytes were also found to be very much increased in diabetics as compared with controls. It was possible that increased expression of HLA-DQ antigen, IL-2R and TfR of monocytes in patients with IDDM might play a role in the pathogenesis of the autoimmune reaction.
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Objective To investigate the association of cytotoxic T lymphocyte associated antigen 4 (CTLA 4) gene polymorphism with type 1diabetesmellitusandautoimmunethyroiddiseases. Methods The A/G phenotype at position 49 of the CTLA 4 gene exon 1 was determined by PCR RFLP method in 33 classic type 1 diabetes patients, 57 latent autoimmune diabetes in adults (LADA) patients, 122 autoimmune thyroid disease patients and 84 healthy control subjects of Chinese Han. Results The frequency of the CTLA 4 G phenotype was significantly higherintype1diabetespatientsthanthatincontrol subjects (55.6% vs 36.9%, P=0.0005). Neither the presence nor the absence of G allele influenced the occurrence of islet autoantibody (ICA) and glutamate decarboxylase antibody (GADA). The strong association of the CTLA 4 G allele with AITDs was showed in our study (66.4% vs 36.9%, P
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Objective To study the relationship between large multifunctional proteasome (LMP) 7 gene polymorphism and susceptibility of type 1 diabetes mellitus (DM). Methods The genotyping of LMP7 gene was determined by polymerase chain reaction restriction fragment length polymorphism (PCR RFLP) in 71 type 1 DM patients and 86 healthy persons (as controls). Furthermore, the type 1 DM patients were divided into 3 groups according to the age of diabetic onset. Group A was ≤14 years, group B 15~30 years, group C≥31 years.Results The frequency of LMP7 B/B was decreased significantly (39% vs 58%, P