Objective: To investigate the apoptosis of renal cells induced by tumor necrosis factor alpha (TNF-α) and nuclear factor-κB (NF-κB) in diabetic rats and intervention of rapamycin. Methods: A total of 20 rats (Goto-Kakizaki rats) with type 2 diabetes mellitus (T2DM) were randomly and equally divided into DM model group (DM group) and rapamycin treatment group (DMR group, received rapamycin treatment after DM model was established); another 10 Wistar male rats were regard as normal control group. Apoptosis of renal cells, expression levels of TNF-α and NF-κB and levels of blood lipids, blood glucose were measured in all groups after four weeks and eight weeks. Results: Four and eight weeks After model was established, compared with normal control group and DMR group, there were significant increase in renal cells apoptosis [RCA, four weeks: (0.217±0.031), (0.272±0.031) vs. (0.545±0.031), eight weeks: (0.358±0.031), (0.350±0.031) vs. (0.811±0.031)] and expressions of NF-κBp65 [OD: four weeks: (0.160±0.027), (0.131±0.027) vs. (0.411±0.027), eight weeks: (0.232±0.027), (0.275±0.027) vs. ( 0.634±0.027)] and TNF-α [OD: four weeks: (0.242±0.027), (0.275±0.027) vs. (0.617±0.027), eight weeks: (0.385±0.027), (0.342±0.027) vs. (0.912±0.027)] in DM group (P<0.01 all). Correlation analysis indicated that there were positive correlations between renal NF-κBp65 and TNF-α, among RCA and TNF-α, NF-κBp65 (r=0.956, 0.953, 0.886，P<0.01 all).

Objective To compare the efficacy of insulin aspart and human soluble insulin on postprandial glucose control and blood glucose excursion in type 2 diabetic patients managed with insulin pump therapy. Methods All of 345 hospitalized type 2 diabetic patients were randomized divided into two groups. One group underwent insulin pump therapy with insulin aspart (aspart group, 173 cases),another group with human soluble insulin (humulin R group, 172 cases). Capillary glucose concentrations were measured at 9 time points,including preprandial,2 hours postprandial,bedtime (22:00),midnight(0:00) and 3:00 every day during the treatment. The change of blood glucose at each time point and the variation of postprandial blood glucose excursion was compared between the two groups. The frequency of hypoglycemia was also evaluated. Results After treatment, fasting blood glucose and post breakfast and post dinner blood glucose levels were decreased more significantly in the aspart group than those in the humulin R group. And a significantly smaller postprandial blood glucose excursion was shown in the aspart group compared with that in the humulin R group (P< 0.05). The time to achieve good glycemic control in the aspart group was (4.40 ± 2.16) d, significantly shorter than that in the humulin R group[(5.68 ± 2.29) d](P< 0.05). The incidence of hypoglycemia was significantly lower in the aspart group (P <0.05). Conclusion Insulin aspart results in better control of blood glucose and less glycemic variability compare with human soluble insulin in type 2 diabetic patients during delivery by continuous subcutaneous insulin infusion.

Objective To explore the application of continuous subcutaneous insulin infusion (CSII) in elderly type 2 diabetic patients. Methods Elderly group(n = 415) and control group (n= 461) received the transient CSII intensive treatment. The different regiments and the incidence of hypoglycemia were observed between the two groups. Results The average duration reaching the target blood glucose and the insulin dosage at the target time were similar between the two groups (P>0.05). The insulin basal rate in the elderly patients was lower than that in control group. The incidence of hypoglycemia, especially at night, was significantly higher in the elderly patients.Conclusions CSII could control blood glucose effectively in elderly type 2 diabetic patients, but the basal insulin dosage should be decreased to reduce the risk of hypoglycemia.

BACKGROUND: Previous studies found that C57BL/6 mouse was susceptible to atherosclerosis, while BALB/c mouse was resistant to atherosclerosis. The stenosis of the culprit vessel and the severity of myocardial infarction were correlated to the levels of OX40L expression. Whether OX40L has differential expression between C57BL/6 and BALB/c mouse was not identified.OBJECTIVE: To observe the differential expression of OX40L mRNA and protein in C57BL/6 and BALB/c mouse.METHODS: Total RNA and protein were extracted by Trizol and RIPA Buffer from heart, brain, kidney, skeletal muscle and spleen tissues of C57BL/6 and BALB/c mice. RT-PCR and Western Blot were used to detect OX40L mRNA and protein expression in heart, brain, kidney, spleen and skeletal muscle of two kinds of mice. The differential expression of OX40L mRNA and protein between C57BL/6 and BALB/c mice was observed.RESULTS AND CONCLUSION: RT-PCR results showed that the mRNA expression level of OX40L in heart of C57BL/6 mice mouse was significantly higher than BALB/c mice (P ＜ 0.05); the mRNA expression level of OX40L in spleen of BALB/c mice was significantly higher than C57BL/6 mice (P ＜ 0.05). There were no significant differences in the brain, kidney and skeletal muscle between these two strains. The results of Western Blot showed that the protein expression level of OX40L in heart, brain and kidney of C57BL/6 mice were significantly higher than BALB/c mice (P ＜ 0.05). There were no significant differences in skeletal muscle and spleen between these two strains. The OX40L mRNA transcription level in heart was higher in C57BL/6 mouse than BALB/c mouse, while the expression in spleen was lower than the latter. The OX40L protein levels in C57BL/6 mouse heart, brainand kidney were higher than BALB/c mouse. The differences of OX40L expression between the two strains of mice indicated that OX40L may promote to C57BL/6 mouse susceptible to atherosclerosis.

BACKGROUND: To apply mouse anti-human cTnI monoclonal antibody as the drug vector in the treatment and diagnosis of myocardial injury, it is important to degrade the immunity of murine antibody and overcome human anti-mouse reaction. Humanization has been applied as an attempt to resolve this problem.OBJECTIVE: To clone murine anti-cTnI Fab fragment and analyse the nucleotide and deduced amino acid sequences.DESIGN: Single sample study.SETTING: An institute of cardiovascular disease under a medical university-affiliated hospitalMATERIALS: The study was conducted in the Institute of Cardiovascular Diseases, First Affiliated Hospital of Nanjing Medical University from January 2003 to May 2004. The hybridoma cell line JS200202 which secrets the anti-cTnI monoclonal antibody was provided by Institute of Cardiovascular Disease, First Affiliated Hospital of Nanjing Medical University.METHODS: IgG heavy chain primers and κ light chain primers of amplified mouse were designed. Total RNA was extracted from hybridoma cells which secrete cTnI. Reverse transcription polymerase chain reaction(RT-PCR) was amplified. Cloning and subsequent sequence analysis of the Fab fragment was performed. The deduced amino acid sequence was compared and analysed with previously published sequences.MAIN OUTCOME MEASURES: Heavy chain Fd segment and κ light chain gene sequence and its subgroups.RESULTS: A band of approximate 700 and 800 base pairs were amplified using IgG heavy chain primers and κ light chain primers respectively. Sequence analysis indicated that the deduced amino acid sequences were in consistent with the characterization of the amino acid in the murine IgGl Fab fragment(GenBank accession NO AY484430, AY484431; Protein Bank accession NO AAR83243, AAR83244).CONCLUSION: A complete murine anti-cTnI Fab fragment was obtained in this study, which may provide basis for the production of the chimeric anti-cTnI antibody.

Objective: To study expressions of small ubiquitin-related modifier protein（SUMO）4 (SUMO4), nuclear factor (NF)- κB and inhibitory factor of NF-κB (IκB) in kidneys of rats with type 2 diabetes mellitus (T2DM). Methods: A total of ten 40-week-old male Goto-Kakizaki (GK) rats （with spontaneous diabetes mellitus）of specific-pathogen free (SPF) grade, and ten 40-week-old male Wistar rats of SPF grade were selected. The lesion of renal tissue was observed by hematoxylin eosin (HE) staining. Expressions of SUMO4, NF-κB and IκB in renal tissue were observed by immunohistochemistry methods. Results: In the GK rats, glomerular capillary ball hypertrophy, basilar membrane slightly thickening; glomerular mesangial cells hyperplasia, hypertrophy and renal tubular epithelial cells hypertrophy were observed. Compared with normal Wistar rats, expression levels of NF-κB [(0.232±0.034) vs. (0.634±0.058)], IκB [(0.242±0.027) vs. (0.712±0.078)] and SUMO4 [(0.160±0.031) vs. (0.545±0.045)] significantly increased in renal tissue of GK rats (P<0.01 all). Conclusion: Compared with Wistar rats, expressions of NF-κB, IκB and SUMO4 significantly increase in renal tissue of GK rats, suggesting that SUMO inhibiting transcriptional activity of NF-κB may exist in kidneys of T2DM rats. Therefore, sumoylation may be a new therapeutic target for inhibit renal microvascular lesion of diabetic disease.

Objective: To study relative risk factors for diabetic nephropathy (DN). Methods: A total of 238 patients diagnosed as type 2 diabetes mellitus (DM) were enrolled in the study. According to urine microalbuminuria to urine creatinine ratio (UACR), patients were divided into pure DM group (group DM1, n=90), early diabetic nephropathy group (group DM2 , n=73) and clinical diabetic nephropathy group (group DM3 ,n=75). Clinic data of all patients were collected; Fasting blood glucose (FBG), 2h postprandial blood glucose (2hPB), blood lipids, uric acid (UA), fibrinogen (Fg) and glycosylated haemoglobin (HbA1c) were measured in all patients, and their correlations with DN were analyzed. Results: Compared with group DM1, the course of disease in DM [(7.25±6.29) years vs. (10.25±7.67) years vs. (13.53±7.82) years], levels of FBG [(8.46±2.52) mmol/L vs. (9.52±3.38) mmol/L vs. (10.82±3.30) mmol/L], 2hPB [(18.40±5.64) mmol/L vs. (20.27±5.94) mmol/L vs. (22.59±6.14) mmol/L], HbA1c [(7.96±1.65) % vs. (8.60±1.76) % vs. (9.55±2.09) %], triglyceride [TG, (1.72±0.86) mmol/L vs. (2.34±1.87) mmol/L vs. (3.16±1.85) mmol/L], Fg [(3.49±0.93) g/L vs. (3.88±1.21) g/L vs. (4.99±2.10) g/L] and UA [(295.42±52.34) μmol/L vs. (324.18±96.29) μmol/L vs. (351.23±56.88) μmol/L] significantly increased in group DM2 and group DM3 in order (P<0.01~0.001). Logistic gradual regression analysis indicated that course of DM, HbA1c, TG, Fg and UA were risk factors for DN (OR=1.008~1.910, P<0.01~0.001). Conclusion: The course of DM, blood glucose, blood lipid, uric acid and fibrinogen are risk factors of diabetic nephropathy; increased UACR reflects progress of patient’ condition in DM patients, its detection is used for diabetic prognosis and treatment.

Objective: To study influence of overweight and obesity on blood coagulation and metabolic disorders in patients with type 2 diabetes mellitus (T2DM). Methods: A total of 248 preliminary diagnosed T2DM patients were selected. According to body mass index (BMI), they were divided into normal BMI control group (n=95), overweight group (n=87) and obesity group (n=66). Blood lipids, blood glucose and fasting insulin (FINS) were measured in all patients and homeostasis model-insulin resistance index (HOMA-IR) was calculated then. Statistical analysis was performed. Results: Compared with normal BMI control group, there were significant increase in fibrinogen [(3.37±0.55) g/L vs. (4.04±0.70) g/L vs. (5.20±0.69) g/L], urine microalbumin [(14.46±8.90) mg/g vs. (47.33±42.54) mg/g vs. (104.45±60.78) mg/g], fasting blood glucose [ (7.15±0.97) mmol/L vs. (8.84±1.81) mmol/L vs. (10.06±2.28) mmol/L], FINS [(10.09±8.21) IU/ml vs. (14.33±15.55) IU/ml vs. (19.69±10.86) IU/ml], HOMA-IR[(3.19±2.59) vs. (5.51±5.38) vs. (8.48±4.62)], TG, TC and LDL-C levels in overweight group and obesity group, and the more BMI patients were, the higher these indicators were; There were significant decrease in plasma prothrombin time [(13.33±0.69)s vs. (12.74±0.69)s vs. (11.43±0.53)s], activated partial thromboplastin time [ (37.32±2.31)s vs. (36.55±2.41)s vs. (34.61±1.53)s] and HDL-C [(1.54±1.12) mmol/L vs. (1.27±0.41) mmol/L vs. (1.09±0.28) mmol/L] in overweight group and obesity group（P<0.05 all）. Conclusions: Overweight and obesity aggravate coagulation and metabolic disorders in patients with type 2 diabetes mellitus. It also aggravates degree of insulin resistance, the more BMI patients are, the more serious they are.

Objective To study the regiments of continuous subcutaneous insulin infusion (CSII) in admitted type 2 diabetic patients, and to analyse the factors related to its effectiveness and insulin dosage. Methods A total of 1 276 type 2 diabetic patients were treated by CSII. The total efficacy of CSII was evaluted. The use of CSII was also analysed in the newly diagnosed patients, elderly patients, and patients with obese or infectious disease. Results The excellent control of blood glucose were achieved in (5.7±2.6)days in the dosage more early and quickly in the newly diagnosed group than that in the previously diagnosed group after the blood glucose levels achieved good control. The percentage of the patients reached the clinical relieve was also higher in the newly diagnosed group. The incidence of hypoglycemia was significantly higher in the elderly patients with lower basal insulin dosage at night. The bolus insulin dosage in the obese patients was higher than that in the non-obese patients. The patients with infectious disease usually have a higher basal insulin dosage than those without infectious disease. The days needed for achieving good control of blood glucose and the insulin dosage were related to infectious factors, the basal blood glucose and obesity. Conclusion The application of CSII among the patients is varied with different conditions. Blood glucose level, body mass index and infection factors are important to determine the initial insulin dosage.

Objective: To analyze the clinical characteristics of X-linked inhibitor of apoptosis (XIAP) deficient patients with clinical manifestation of Crohn's disease. Methods: Clinical manifestations, laboratory investigations, genetic testing and therapeutic interventions of one case of XIAP deficiency who was admitted to Department of Gastroenterology in Children's Hospital, Zhejiang University School of Medicine in May 2016 were summarized. PubMed and Chinese database for articles published from January 2016 to June 2017 were searched using the key words of'Crohn's disease’and'XIAP’, and the relevant literature was reviewed. Results: The case we reported was a 6-year-1-month-old boy with recurrent bloody stool for 2 months, and abdominal pain with fever for 2 weeks. The patient had a past history of hemophagocytic lymphohistiocytosis (HLH) and epilepsy in the past one year. Complete blood cell count showed mild anemia (Hb108 g/L). The patient had an elevated high-sensitivity C reactive protein (86 mg/L) and erythrocyte sedimentation rate (46 mm/1h) . White blood cells, pus cells and red blood cells were found on routine stool examination. Biochemical panel showed hypoalbuminemia (25.2 g/L) , elevated transaminase (alanine aminotransferase 175 U/L, aspartate transaminase 229 U/L) , hypertriglyceridemia (4.41 mmol/L) , and hyperferritinemia (>1 650.0 μg/L) . Magnetic resonance enterography revealed the intestinal wall thickening and increased enhancement in parts of illeum and colon. Capsule endoscopy revealed multiple ulcers in jejunum. Colonoscopy showed multiple ulcers in colon and the pathological examination revealed chronic inflammation in mucosa of terminal ileum and colon, which was combined with partial necrosis and ulceration. Some phagocytes were seen in bone marrow smears. The patient was given multiple diagnoses, including hemophagocytic lymphohistiocytosis, Crohn's disease, sepsis, epilepsy, severe malnutrition, and hypoproteinemia. The pediatric Crohn's disease activity index (PCDAI) was 37.5. Genetic testing identified a hemizygotic mutation of c.910G>T chrX:123022501 p.G304X in XIAP. The parents had no such mutation. The patient showed response to infliximab with oral intake of mercaptopurine and corticosteroids, and had remission with PCDAI of 0. There were 9 relevant articles (Chinese 0 English 9), which showed 33.3% XIAP deficient patients manifested with inflammatory bowel disease(IBD), who might have other manifestations such as hemophagocytic lymphohistiocytosis or splenomegaly simultaneously or sequentially. Those patients showed poor response to monotherapy. Conclusion XIAP deficient patients have various clinical manifestations. Genetic testing is important to those male pediatric IBD patients who have the complicated symptoms or little response to standard therapy.