Objective To observe the effect and safety of the human glucagon-like peptide-1 analogue,liraglutide,versus insulin glargine in patients with type 2 diabetes mellitus inadequately controlled with metformin alone.Method Ninty patients with type 2 diabetes mellitus(aged 18-79 years,HbA1C 7.5％-10.0％,body mass index＜40 kg/m2) who had inadequate glycaemic control on metformin were allocated for the research with an open,randomized,parallel controlled clinical research method.The patients kept the original dose of metformin unchanged and were randomly assigned to the liraglutide group or the insulin glargine group according to a proportion of 1 ∶ 1.Liraglutide group started with a dose of 0.6 mg subcutaneous injection qd,changed to 1.2 mg subcutaneous injection qd after one week and kept unchanged until the end of the research.Insulin glargine group started with a dose of 0.1-0.2 U/kg according to the fingertips peripheral blood glucose level before breakfast on the continuous 3 d before every follow-up.At the baseline,after 4 weeks,12 weeks,20 weeks,and 26 weeks of treatment,HbA1C,blood glucose,lipids weight,blood pressure were arranged to measured.86 patients finally completed the study.Results Mean HbA1C and the success rate of HbA1C ＜7％ were similar between liraglutide group and insulin glargine group [(7.06 ± 0.87) ％ vs (7.25 ± 1.20) ％,47.73 ％ vs 45.23 ％,P＞0.05],while the percentage of subjects reaching the composite endpoint of HbA1C＜7％ with no hypoglycemia and no weight gain was significantly higher in liraglutide group than insulin group(P＜0.05) ; Fasting plasma glucose decreased more markedly in insulin glargine group,2 h postprandial plasma glucose was decreased more markedly in liraglutide group(P＜0.05 or P＜0.01).Liraglutide significantly reduced mean body weight by (3.21 ± 1.18) kg,waist circumference by (3.82 ± 1.21) cm,and body mass index by (1.95 ± 0.61) kg/m2 (P＜0.01 or P＜0.05),while in the insulin glargine group there sere rise of respective figure of(2.86 ± 0.43) kg,(1.52 ± 0.56) cm,and (0.61 ± 0.25) kg/m2 (P＜0.05),systolic blood pressure and serum triglyceride declined.There was no serious adverse affect in both groups,the incidence of mild hypoglycemia was significantly less in liraglutide group and has a statistically significant difference (4.55％ vs 21.43％,P＜0.05).Conclusions Liraglutide showed a good effect on reducing weight,systolic blood pressure,blood lipid and in addition to blood glucose control which is comparable to insulin glargine.What is more,liraglutide had good safety and tolerability,which can be regarded as a good choice for patients with type 2 diabetes mellitus inadequately controlled with metformin alone.

Simultaneous concurrence of subacute thyroiditis and Graves' disease is rare.We present one case of subacute thyroiditis with Graves' disease and combine with other reports to explore the clinical characteristics and therapeutic considerations.If subacute thyroiditis is considered coexisting simultaneously with Graves' disease,radioactive iodine uptake,thyroid autoantibody,fine-needle aspiration of thyroid gland,thyroid nuclide imaging examination,etc,should be done to make correct diagnosis and to adjust the therapeutic plan.

Objective To observe the curative effect on non-obese type 2 diabetes and the effect on change of glucagon-like peptide-1 (GLP-1) of gastric bypass operation. Methods Thirty-two cases of gastric ulcer with non-obese type 2 diabetes were suffered gastric bypass operation. Plasma glucose concentrations, insulin and GLP-1 were measured respectively in fasting and postprandial conditions before operation and in week 1, 2, 3 and month 1, 3, 6 after gastric bypass operation, and the body mass index (BMI), homeostasis model assessment ? cell function index (HBCI) and glycosylated hemoglobin (HbA1c, the index was detected only before operation and in month 3, 6 after operation) were also measured. The turnover of the diabetes condition in the 6th month after surgery was observed. Results Compared with the levels before operation, the fasting and postprandial plasma glucose levels were descending (P0.05). The diabetes control rate was 78.1%(25/32) overall six months after operation. Level of GLP-1 was negatively correlated with level of plasma glucose (P

Objective To investigate the effect of gastric bypass on plasma glucose and glucagonlike peptide-1 (GLP-1) during oral glucose tolerance test (OGTT) in non-obese type 2 diabetes mellitus (T2DM) patients suffering from gastric ulcer necessitating a gastrectomy.Methods Thirty-two T2DM patients undergoing gastric bypass (GBP) for gastric ulcer were enrolled in this study.All patients underwent a 3-h OGTT with 75 g glucose (in a total volume of 300 ml) preoperatively and 1 week,1 month,3 months and 6 months after operation.Plasma glucose (PG) and GLP-1 concentrations were measured before (baseline) and then 30,60,120,and 180 minutes after OGTT.Areas under curves (AUC) were calculated by trapezoidal integration.The turnover of the diabetes conditions six months after the surgery were also measured.Results FPG level was (9.5±1.0) mmol/L before surgery,and significantly decreased to (7.4±1.0) mmol/L,one week,(6.5±1.2) mmol/L,one month,(8.0±1.6) mmol/L three months and (5.8±1.0) mmol/L,six months respectively after GBP(P ＜0.01).Peak level and AUC of PG during OGTT significantly decreased at aforementioned four time points after sugery (P ＜ 0.01).Oral glucosestimulated peak levels of GLP-1 was (20±3) pmol/L before GBP,and markedly increased to (83±15)pmol/L,(86±20) pmol/L,(87±22) pmol/L and (92±20) pmol/L respectively after GBP(P＜0.01).Stimulated AUC levels of GLP-1 significantly increased from (2457±395) pmol· min/L to (6499±1227)pmol·min/L,to (7275±1475) pmol·min/L,to (7307±1575) pmol·min/L and to (7974±1594)pmol· min/L during the study respectively(P ＜0.01).BMI levels were similar before and after GBP(P＞0.05).Two patients experienced complications(infection of incision and intractable hiccup).T2DM control rate was 78% when assessed at a time point of six months later.Conclusion Gastric bypass is effective in terms of glucose control and improving gucose tolerance in non-obese T2DM,and the hypoglycemic effect may be contributed to more GLP-1 secretion after GBP,but not to weight loss.

Objective To investigate the relationship between recent chlamydia pneumoniae (Cp)infection and active ankylosing spondylitis (AS). Methods Seventy nine AS outpatients and 73 normal controls (NC) were enrolled into this study. Serum anti-Cp antibodies (CpIg) were tested using the enzymelinked immunosorbent assay (ELISA). Clinical and experimental data were collected. Patients with positive CpIgM or CpIgA were considered as having a recent Cp infection. Wilcoxon test, Student's t test, χ2 test and multivariate logistic regression were used for statistical analysis. Results Both AS patients and normal controls had a high prevalence for sero-positive CpIgG, which was 89％(70/79) vs 92％(67/73) respectively,while AS patients had a higher frequency of CpIgA and CpIgM when compared with NC [52％(41/79) vs 32％(23/73), χ2=6.61, P=0.010 for CpIgA; 80％(63/79) vs 21％(15/73), χ2=44.031, P＜0.01 for CpIgM]. The presence of CpIgM or CpIgA favored AS, the OR was 17.1 (95％CI 7.4～39.5), or 3.1 (95％CI 1.3～7.2),respectively. In addition, CpIgM was associated with disease activity parameters including ESR (χ2=2.56, P=0.021), CRP (χ2=7.28, P=0.007) and BASDAI (χ2=6.79, P=0.009). Furthermore, consecutive positive CpIgM favored the persistent active or relapsed disease, while negative CpIgM favored a reduced disease activity.There was no correlation between CpIgM/CpIgA and peripheral joint disease and enthesitis. Conclusion Recent Cp infection is highly associated with AS and CpIgM antibody relates with active AS, which indicates that Cp infections may be a critical triggering factor for active AS.

BACKGROUND: Angiotensinogen (AGT) gene is the firstly discovered candidate gene for essential hypertension, both the T174M and M235T polymorphisms locate at the second exons of AGT gene, and there is existence of linkage disequilibrium. The polymorphism at A-6G and G-217A sites in promotor region plays an important role in regulating the gene expression, and the products of keep close correlation with the level of blood pressure. OBJECTIVE: To investigate the association between the polymorphism of AGT gene at A-6G, T174M and G-217A sites and the risk for the attack of essential hypertension in Chinese Han population, DESIGN: A cluster sampling and case-control analysis. SETTINGS: Department of Geriatrics and Department of Cardiology, the First Affiliated Hospital of Nanjing Medical University; Southern Research Center of National Human genome; Department of Cardiology, Dongtai People's Hospital of Jiangsu Province. PARTICIPANTS: The experiment was carried out in the countryside of Dongtai county, Yancheng city, Jiangsu province. All the subjects were selected from the countryside of Dongtai county, Yancheng city, Jiangsu province. Totally 177 patients with essential hypertension who had never accepted any drug treatment, were taken as the essential hypertension group, and hypertension was diagnosed according to the diagnostic standard of hypertension set by WHO/ISH in 1999 (systolic blood pressure ≥ 140 mm Hg and/or diastolic blood pressure ≥ 90 mm Hg); Another 86 normal person were taken as the normal control group. ② Inclusive criteria: The enrolled subjects should be Han nationality; long-term local residents but not from other places; able to answer questions clearly; diagnosed by disease history, clinical symptoms, physical signs and assistant examinations; have complete data of investigation of uniform questionnaires by face-to-face interview (including demographic information, profession history, family history and life styles of smoking, drinking, drinking tea, etc.). ③ Exclusive criteria: The patients with secondary hypertension in the essential hypertension group, subjects having family hisory of hypertension in the normal control group, and those with chronic diseases of liver and kidney, and diabetes mellitus in both groups were excluded. METHODS: Peripheral venous blood samples (3 mL) were collected, and DNA was extracted from human peripheral blood with FlexiGene DNA Kit (250). The Primer3 software was applied to design primers, and the polymorphism sites in the primer sequence were excluded. After multiplex polymerase chain reaction (PCR), 3 μL products were selected to detected the amplified results by agarose gel electrophoresis. The successfully amplified PCR products were purified with the QIAquick PCR Purification Kit, and the purified products were fragmentized with Dnase Ⅰ . The fragmentized products of enzyme digestion were labeled with fluorescein by deoxynucleotide terminal transferase. Two allele specific probes and one mismatched probe were designed respectively for each single nucleotide polymorphism. The chips were prepared with the OmniGridTM 100 TLC samler, each probe was repeated for three times to form three matrix. The hyridization solution was degenerated at 95 ℃ for 10 minutes, and then immediately cut on ice. 10 μL hybridization solution was added onto the chip matrix, hybridized at 50 ℃ for 2 hours, then washed and dried. The chips were scanned with the GenePix 4000B laser confocal scanner (Figure 2),and the intensity of the fluorescent signal for each probe was extracted with GenePix Pro, and the allele score of each single nucleotide polymorphism was calculated to judge the genotype. MAIN OUTCOME MEASURES: ① Comparison of the frequencies of genotype distribution at each polymorphism site of AGT gene in both groups; ② Correlation analysis of the polymorphism of AGT gene at A-6G and T-174M sites with the risk for the attack of essential hypertension; ③ Effects of the polymorphism of AGT gene at A-6G, T-174M and G-217A sites on blood pressure.RESULTS: According to the intention-to-treat analysis,all the 263 subjects were involved in the analysis of results. ① At the A-6G site of AGT gene, the frequencies of AA, AG and GG genotypes (P=0.014) and A and G alleles (P=0.004, OR=0.44) had significant differences between the essential hypertension group and normal control group; At the T174M site, the frequencies of CC, CT and TT genotypes (P=0.031) and A and G alleles (P=0.014, OR=0.55) were significantly different; At the G-217A site, no obvious differences were found in the GG, AG and AA genotypes (P=0.722) and G and A alleles (P=0.403, OR=0.80). ② The risk of essential hypertension in the individuals carrying AA genotype of A-6G polymorphism and CC genotype of T174M polymorphism was reduced by 57% (95%CI= 0.23-0.82, P= 0.010) and 56% (95%CI= 0.25-0.79, P= 0.006) respectively. ③ There were no significant differences in the systolic blood pressure, diastolic blood pressure and mean arterial pressure among different genotypes at the A-6G, T174M sites and G-217A sites (F=0.100- 2.911, P ＞ 0.05). CONCLUSION: The AA genope at A-6G and the CC genotype at T174M site of AGT gene may reduce the risk for the attack of essential hypertension in Chinese Hun population, and no significant correlation was found between the genotype of G-217A polymorphism and the attack of essential hypertension.

Objective The truncated fibronectin-binding proteins A (Fba protein) were cloned and expressed, then animals were immunized with Fba protein and subsequently challenged with group A streptococcus (GAS) to further investigate protective antibodies induced by each domain and determine the most immunogenic domain. Methods Fba proteins, which were divided into four overlaps based on the structural domains, were truncated and expressed. The fba truncated genes were amplified by polymerase chain reaction (PCR) with SSI-9 of GAS as template, and cloned into prokaryotic expression plasmid pGEX-2T and expressed in E. coli BL-21. The products were confirmed by Western blot and purified by affinity chromatography column. Female BALB/c mice were immunized with the four truncated proteins respectively, with phosphate buffered solution (PBS) as control. The serum IgG of mice was detected by enzyme-linked immunosorbent assay (ELISA). After the third immunization, mice were challenged with fatal dose of GAS (M+ Fba+ ) to evaluate the protective rates in each group. The data were compared by analysis of variance and Fisher's exact test.Results Prokaryotic expression plasmids of pGEX-2T/FbaAl, pGEX-2T/FbaA2, pGEX-2T/FbaA3 and pGEX-2T/FbaA4 were successfully constructed and the four truncated proteins were expressed and purified successfully. Serum levels of IgG in each experimental group gradually increased with immunization with Fba protein more times. After the third immunization, the IgG titer against FbaA2 1290.2, P<0. 01). After GAS challenge, four out of eight mice were protected in FbaA2 protein group, while two out of eight mice in FbaAl protein, FbaA3 protein and FbaA4 protein groups,respectively (P<0. 05). Conclusions Four truncated Fba proteins are constructed and expressed successfully. Truncated FbaA2 protein could be able to induce strongest protective immune response.

Objective:To identify monoclonal antibody McAb2 recognizing epitope of Fba of GAS.Methods:The overlapped peptides were synthesized and their abilities to bind McAb2 were detected by dot-ELISA.The predominance amino acids specific for McAb2 were screened using phage 7 peptide library.Results:The result by dot-ELISA analysis demonstrated that the synthetized peptide,amino-acid residues 100-112~(th),could bind McAb2 with high affinity.The predominance amino acids specific for McAb2 were ITPDL,which was located in 100-110~(th)aa of Fba by panning with phage 7 peptide library.Conclusion:The domain and the predominance amino acids of Fba recognized by McAb2 is determined.The results would contribute to the research of the role of Fba on the pathogenic mechanism of GAS,the identification of function of McAb2,and the development of epitope-peptide vaccine.

The effects of repaglinide combined with glargine (n=31)on glucose metabolism and β-cell function were observed in the patients with type 2 diabetes after secondary sulfonylureas failure and the results were compared with glimepiride combined with glargine (n=32). The preprandial capillary blood glucose, postprandial capillary blood glucose and HbA1C in both groups after 6-month treatment were significantly reduced as compared with those at baseline (all P＜0.01). The treatment with repaglinide(2 mg tid) plus glargine was more efficient than glimepiride(4 mg qd) plus glargine in improving β-cell function, ameliorating HbA1C and postprandial blood glucose excursions in patients with type 2 diabetes.

Objective To discuss effectiveness of gracilis muscle flap for treatment of degree 3 and 4 pressure ulcers on ischial tuberosities for elderly patients.Methods From January 2010 to June 2015,19 elderly patients with pressure ulcers on the ischial tuberosities were enrolled,including 11 males and 8 females with an average age of 71.3 years ranging 62 to 86 years.The average course of disease was 9.6 months ranging 2 to 23 months,including 7 cases of degree 3 and 12 cases of degree 4 according to the National Pressure Ulcer Advisory Panel (NPUAP).All cases were treated by gracilis musle flap after debridement.Results All muscle flaps survived without necrosis,2 cases had wound dehiscence after operation and were healed after symptomatic treatment,and the 17 wounds were healed for first intention.All patients were followed up for 6 months to 5 years.Pressure ulcer on the ischial tuberosities recurred in 1 case after operation and was healed by debridement and suture.Pressure ulcers did not recur in the rest cases.Conclusions Gracilis muscle flap in the treatment of degree 3 and 4 pressure ulcers on the ischial tuberosities has remarkable curative effect and high cure rate in elderly patients,it is worthy of clinical application.