Objective To observe the influence of short-term continuous subcutaneous insulin infusion(CSII)on ?-cell function in newly diagnosed type 2 diabetes(T2DM).Methods A total of 32 inpatients with newly diagnosed T2DM who had a 14-day course of CSII were randomly assigned to two groups of OGTT and arginine stimulating test(AST).Before and after 3 days and 14 days CSII treatments,OGTT and AST were performed respectively.The changes of the values of plasma glucose,HOMA-IR,HOMA-? and △C30/△G30,△C120/△G120 or arginine-induced acute C-peptide reaction(ACRARG)in two groups were compared.Results In OGTT,the values of plasma glucose and HOMA-IR were decreased,HOMA-? and △C120/△G120 were increased significantly after CSII therapy,both in day 3 and day 14;and △C30/△G30 were unchanged after 3 days but increased markedly after 14 days.However,in ACRARG,the above mentioned parameters almost remained unchanged after CSII treatment in 3 days or 14 days.Conclusions In newly diagnosed T2DM with elevated fasting glucose levels,intensive insulin therapy can partly improve ?-cell function showed by OGTT,not by AST.

Objective To investigate the clinic effect of trans-artery intrahepatic transplantation of porcine islet in type 1 diabetic patients. Methods Four patients with type 1 diabetes were perfused newborn porcine islets through the hepatic artery.Fasting blood glucose(FBS), postprandial blood glucose (PBS), HBA1,dose of insulin and function of liver and kidney were measured before and after the transplantation.Results The levels of FBS,PBS and HBA1 were 7 4～13 5mmol/L,7 8～18 6mmol/L and 7 8～12 0mmol/L respectively before the transplantation. During one and half years after transplantation, the levels of FBS,PBS and HBA1 were 3 3～7 1mmol/L,6 0～8 8mmol/L and 6 4～7 6mmol/L respectively with notably reduced dosage of insulin(32～58%).The function of liver and kidney of the patients remained normal.Conclusions Trans-artery intrahepatic transplantation of porcine islet is effective and safe for type 1diabetic treatment.

Objective: To investigate the effects of glucagon-like peptide-1(GLP-1)on high glucose-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the mechanism involved. Methods: HUVECs were cultured under varying conditions for 48 h, and the cell viability was spectrophotometrically measured by MTT assay. Flow cytometry detected the ratio of cell apoptosis. Western blot detected the protein levels of p-Akt and p-eNOS, while NO assay kit detected the NO concentration.
Results: Treatment of high glucose (33 mmol/L) for 48 h signiifcantly decreased the HUVECs viability and induced the apoptosis of HUVECs, concomitant with decreased Akt and eNOS phosphorylation leves and subsequent NO production. Treatment with GLP-1 (3 nmol/L) for 48 h in the high glucose group increased the HUVECs viability (P<0.01), decreased the ratio of HUVECs early apoptosis (P<0.05), ameliorated the reduced protein levels of p-Akt and p-eNOS caused by high glucose, and increased the NO production (P<0.05). The anti-apoptotic effect and the increased NO production of GLP-1were inhibited by PI3K inhibitor wortmannine (100 nmol/L) or eNOS inhibitor L-NAME (100μmol/L). The effect on p-Akt, p-eNOS of GLP-1 was inhibited by wortmannine (100 nmol/L) while L-NAME (100μmol/L) did not have any influence on the expression of p-Akt.
Conclusion: GLP-1 can ameliorate high glucose-induced HUVECs apoptosis, which is probably related to the up-regulation of PI3K/Akt/eNOS pathway.

Objective To investigate the effect of erythropoietin on the proliferation,differentiation,and apoptosis of the cultured neonatal porcine islet cells in vitro.Methods Neonatal porcine islet cells were separated and pured from neonatal pigs with collagenase digestion and tissue culture,and their viability and purity were tested. The neonatal porcine islet cells were divided into a control group and an experimental group.The experimental group was treated with erythropoietin but not the control group,and the insulin secretion responsiveness induced by low and high glucose stimulation in the islet was tested after 5 days. Cells were counted and the activation of amplification was determined by MTT chromatometry. The rates of cell apoptosis were observed by ethidium bromide/acridine orange (EB/AO) of fluorescent light staining and flow cytometry,and the cell cycle was analyzed by flow cytometry. The expression of bcl-2,bax,caspase-3,glucose transporter 2 (GlUT-2),and pancreatic duodenal homeobox-1 (PDX-1) mRNA was tested by RT-PCR.Results After erythropoietin was treated in the cell culture,the neonatal porcine islet cells had normal morphology,function,and reaction of insulin secretion to the glucose stimulation. Cell count showed more cells in the experimental group than in the control group (P＜0.05). MTT chromatometry showed the optical absorbance tended to increase with time,and compared with the control group,the optical absorbance was higher in the experimental group (P＜0.05),the expression of PDX-1 mRNA was slightly up-regulated (P＜0.05). The expression of GLUT-2 mRNA had no difference in the 2 groups (P=0.34). In the experimental group,the apoptisis rate was lower than that in the control group by flow cytometry and EB/AO fluoscence staining (P＜0.01),and the expression of bcl-2 mRNA was higher. Howerer bax mRNA and caspase-3 mRNA were obviously lower than those in the control group (P＜0.01).Conclusion Erythropoietin can promote the proliferation but has no effect on the function of neonatal porcine islet cells in vitro. Erythropoietin can protect neonatal porcine islet cells from apoptosis through up-regulating bcl-2 mRNA and downreguling bax and caspase-3 mRNA.

OBJECTIVE To investigate ?-lactamase coding genes and oprD2 gene in multidrug-resistant Pseudomonas aeruginosa isolated.METHODS Polymerase chain reaction(PCR) was used to detect various ?-lactamase coding genes including TEM,SHV,OXA,PER,GES,IMP,VIM,plasmid type AmpC ?-lactamase DHA,MIR and oprD2 in 20 strains of P.aeruginosa.RESULTS The detection rate of ?-lactamase coding genes plasmid type AmpC,VIM,TEM,SHV,OXA and IMP were 15%,5%,15%,10%,10% and 5%,respectively,the loss rate of oprD2 was 25%,but PER and GES genes were negative.CONCLUSIONS The study indicated that the P.aeruginosa is carrying genes of DHA,TEM,SHV,OXA,VIM,IMP and being lost oprD2 gene,the latter is the essential resistance mechanism of P.aeruginosa to beta-lactam antibiotics in local area.

AIM: Some studies found that the glucose uptake ability of diabetes mellitus mice is stronger after feeding with taurine. Whether the taurine can affect adipocytes or not deserves further studies. This article investigates the effects of taurine on the differentiation of 3T3-L1 preadipocytes and detects the mechanics. METHODS: Experiments were performed in the Laboratory Center of Xiangya Third Hospital of Central South University from July to September 2006. ①3T3L1 preadipocytes were provided by Shanghai Cell Bank of China Academy of Sciences. ②3T3-L1 preadipocytes were cultured in high glucose DMEM medium containing fetal bovine serum of 0.10 volume fraction, 108 u/L benzylpenicillin and 80?1010 U/L streptaquaine. After confluence, cells at 5?107 L-1 were incubated in culture flask, and induced with 0.5 mmol/L IBMX, 0.5 mg/L insulin and 1 ?mol/L dexamethasone. The cells in an experimental group were intervened with taurine, whereas these in a control group were not treated with taurine. Oil O staining was used to observe the development of adipose cells. C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours. RNA and protein were extracted in the control group and the experimental group. ③Genes related to adipose cells development was examined by RT-PCR and Western-blot. RESULTS: ①The number of cells stained by oil O was less in the experimental group than the control group in 3T3-L1 preadipocytes treated with 10 mmol/L taurine for 14 days. ②After C3T3-L1 preadipocytes were treated with 10,20 mmol/L taurine respectively for 24 and 48 hours, there were no changes in expression of Insig-2 protein, PPAR, Insing-2, adiponectin, adiponectin receptor, GLUT-4, AP-2 mRNA. CONCLUSION: Taurine inhibits the differentiation of 3T3-L1 preadipocytes, but the mechanics still need to be investigated.

Objective To establish the cell line stably expressing INSIG2 and observe its effecet on fat metabolism after overexpression of INSIG2.Methods The eukaryotic expression plasmid pcDNA3.1(+)-INSIG2 was constructed,which was transfected into 3T3-L1 cells.The expression of INSIG2 and related genes were detected by RT-PCR and immunohistochemistry,the contents of FFA in cell culture medium and adipocyte differentiation were detected by ELISA and Oil Red "O"staining respectively.Results After pcDNA3.1(+)-INSIG2 was transfected into the 3T3-L1 cells,the expression of INSIG1 mRNA and FAS mRNA were down-regulated,the content of FFA in the cell culture medium was decreased and adipocyte differentiation was drepressed.Conclusion The cell line stably expressing INSIG2 was successfully established,the transfected INSIG2 may have a drepressant effect on fat metabolism.

ObjectiveTo investigate the genotype of a case with abnormal hemoglobin combined with hereditary persistence of fetal hemoglobin (HPFH).Methods Male patient,26 years old,were suspected abnormal hemoglobin combined with HPFH after receiving medical examination including hematology exmination,hemoglobin electrophoresis,erythrocyte osmotic fragility analysis in Guangzhou Kingmed Diagnostics in September 2010.Routine examination of anemia and hemoglobin electrophoresis at alkaine pH on agarose gels were applied to analyze the phenotype.Direct sequencing of the complete HBB gene was utilized to identify the hemoglobin variant.Multiplex ligation-dependent probe amplification (MLPA) assay was used to identify the presence of β-globin gene cluster deletion.Gap polymerase chain reaction (gap-PCR) was used to amplify the HBB gene fragment across the breakpoint,and the deletion breakpoint was characterized by direct sequencing the gap-PCR product and comparing the sequencing result with the reference sequence NC_000011.9.ResultsBy direct sequencing of the complete HBB gene,the patient in this study was found to carry a hemoglobin Ta-Li (HBB:c.250G ＞ T) mutation.By combining use of MLPA and gap-PCR with gene sequencing,we found that it had a gross deletion in the β-globin gene cluster,the deletion region was NC_000011.9:g.5222878_5250288del.Therefore,the genotype of this subject was SEA-HPFH combined with abnormal hemoglobin Ta-li.ConclusionCombining application of MLPA and gap-PCR with gene sequencing can help to make sure the genotype.

Objective:To investigate the serum hepcidin level and risk factors associated with peripheral arterial disease (PAD) and foot ulcer in type 2 diabetic patients.Methods:From January 2019 to June 2019, 70 patients with type 2 diabetes in Department of Endocrinology of Xiangya Third Hospital were selected, including 21 newly diagnosed patients with type 2 diabetes (DM group), 23 patients with lower extremity vascular disease (PAD group) and 26 patients with foot ulcer (DF group). Serum hepcidin was determined by enzyme linked immunosorbent assay (ELISA). The serum levels of hepcidin in different groups were compared, and the correlation between diabetic lower extremity vascular disease and foot ulcer was analyzed.Results:⑴ The hemoglobin, albumin, triglycerides and total cholesterol were significantly lower in DF group compared with PAD and DM groups ( P<0.05), while the DF group patients were with higher white blood cell (WBC) count and high sensitivity C reactive protein (hs-CRP) than patients in PAD and DM groups ( P<0.05). DF group also showed significantly higher WBC, hs-CRP and neutrophil ratio level (NEUT%) than DM group ( P<0.05). The inflammatory indicators of WBC, hs-CRP and NEUT% showed no significant difference between DM group and PAD group ( P>0.05). ⑵ The levels of hepcidin in DF and PAD groups were higher than those in DM group, while that in DF group were higher than those in PAD group ( P<0.05); Hepcidin was positively correlated with systolic blood pressure, WBC count, NEUT% and ferritin ( P<0.05), and negatively correlated with hemoglobin, glycosylated hemoglobin, albumin and 25-hydroxyvitamin D ( P<0.05). ⑶ Binary multivariate logistic regression analysis showed that elevated hepcidin level was an independent risk factor for diabetic foot ulcer [ OR=1.755, 95% CI: 1.063-2.897, P=0.028]. Conclusions:The fluctuation of serum hepcidin level in diabetic patients is related to the stimulation of inflammation, the degree of anemia and the nutritional status, which means it might be an early indicator of inflammation in diabetic patients with peripheral arterial disease. Moreover, the increase of hepcidin is an independent risk factor for diabetic foot ulcers in our study.

Type 1 diabetes mellitus and autoimmune thyroid disorders are the most common combination of autoimmune polyendocrine syndrome type Ⅲ(APS Ⅲ). However, APS Ⅲ combined with myasthenia gravis is rare. We described a male patient with myasthenia gravis, type 1 diabetes mellitus, and Hashimoto thyroiditis, who was diagnosed as APS Ⅲ. The human leukocyte antigen (HLA)type was analyzed in this patient. We subsequently reviewed 11 cases of APS Ⅲ combined with myasthenia gravis. This review revealed that HLA-DR9/DQ9 might be a specific HLA subtype associated with APS Ⅲ and complicated with myasthenia gravis .