AIM: To set the model of rat bone marrow me se nchymal stem cells (BMMSCs) that differentiate into cardiomyocytes and observe t he connection structure between cells. METHODS: BMMSCs were isolated by adhering to culture plates and cultured in vitro to expand. Induction of BMMSCs to differentiate into cardi omyo cytes was conducted by treating the cells with 5-aza. Immunocytochemical stainin g was used to identify sarcomeric actin and intercalated disc-like structure wh en the cells were cultured for additional 1 week, 2 or 3 weeks, respectively. RESULTS: Sarcomeric actin positive cells were observed in 1 week , 2 or 3 weeks after 5-aza treatment. Some cells stained positive for connexin4 3 at 1 week and 2 weeks after 5-aza treatment, with brown pellet located disper sively around nucleus. 3 weeks after 5-aza treatment, connexin43 positive cells s howed brown pellet arranging in thready structure around nucleus, such structure could be seen between very few cells which was similar to intercalated disc-lik e structure in normal heart tissue. CONCLUSION: During the process of BMMSCs differentiating into ca rdiomyocytes, intercalated disc-like structure is gradually formed with increas e in culture time and cell density.

Objective To investigate the value of brain natriuretic peptide (BNP) detection in the diagnosis and treatment of heart failure patients with normal ejection fraction.Methods Totally 78 elderly patient were selected in our hospital,with left ventricular ejection fraction (LVEF) ≥45 ％,among which 52 cases of patients were consistent of heart failure criteria (heart failure group),27 cases with normal left ventricular diastolic function as control group.The echocardiographic indices of diastolic function and the change of the concentration of BNP were compared.Results As compared with heart failure group,BNP concentration (108.7 ± 32.2) ng/L vs.(190.3 ± 41.5) ng/L,left ventricular posterior wall thickness (11.3 ± 1.7) mm vs.(13.6 ± 1.4) mm,left ventricular mass index (119.3±10.2)g/m2 vs.(130.7±8.9)g/m2 were elevated in heart failure group (all P＜0.01).Conclusions BNP detection can be used for a diagnosis of heart failure as a simple and easy method.The BNP and ultrasound heart beat graph combination can improve heart failure diagnostic accuracywith normal ejection fraction in elderly patients.

Objective To explore the changes and clinical significance of plasma redox status in patients with acute myocardial infarction,angina pectoris and people with normal coronary artery.Methods According to the clinical manifestation,electrocardiograms and the myocardium markers,68 acute myocardial infarction patients (group A) were involved.68 angina pectoris patients (group B) and 68 normal coronary artery people (group C) were also chosen according to the coronary artery radiographs.Peripheral venous blood of 3 groups were collected.Plasma glutathione (reduced form GSH and oxidized form GSSG) and nicotinamide adenine dinucleotide phosphate (reduced form NADPH and oxidized form NADP+) were detected.The GSH/GSSG and NADPH/NADP+ redox potentials were calculated according to Nernst equation.Results Along with the pathological aggravation of coronary artery (from group C to group A),the levels of GSH [(8.39±1.03)μmol/L,(6.54±0.94) μmol/L,(4.49±0.86) μmol/L,respectively] and GSH/GSSG (14.22±2.14,9.76±1.67,5.76±1.18,respectively) were gradually reduced; the levels of GSSG [(0.59±0.03) μmol/L,(0.67±0.04) μmol/L,(0.78 ± 0.05) μmol/L,respectively] and GSH/GSSG redox potential [(-150.43±3.43) mV,(-141.22±3.12) mV,(-135.21±2.31) mV,respectively] were gradually increased (all P＜ 0.05); the changes of NADPH/NADP+ redox status were similar to GSH/GSSG but milder.Conclusions The imbalance of plasma redox status deviating to oxidation has a relationship with atheromatous plaque formation,plaque rupture and plaque thrombosis in the coronary artery.

Objective To explore the relationship of neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) with gastrointestinal bleeding in adult patients with Henoch-Sch(o)nlein purpura (HSP).Methods Laboratory data including routine blood test results,C-reactive protein (CRP) levels and erythrocyte sedimentation rate (ESR) were collected from 115 adult patients with HSP and 100 healthy controls,and retrospectively analyzed.NLR and PLR were calculated and compared.Logistic regression analysis was used to determine variables associated with gastrointestinal bleeding in adult patients with HSP,and the receiver operating characteristic (ROC) curve was drawn to estimate the predictive efficacy of these variables.Results NLR and PLR were both significantly higher in adult patients with HSP than in healthy controls (both P ＜ 0.05),and NLR,PLR and CRP levels were all significantly higher in adult HSP patients with gastrointestinal bleeding than in those without (all P ＜ 0.05).Logistic regression analysis showed that NLR was an independent risk factor for gastrointestinal bleeding in adult HSP patients (P ＜ 0.05).The ROC curve revealed that the optimal cut-off point of NLR for predicting gastrointestinal bleeding was 3.24 in adult patients with HSP,with the sensitivity and specificity being 85.1％ and 71.3％ respectively.Conclusions Both NLR and PLR evidently increase in adult patients with HSP,especially in those with gastrointestinal bleeding.NLR is a potential predictor for gastrointestinal bleeding in adult patients with HSP.

Objective To investigate the role of Janus kinese 2-signal transducer and activator of transcription 3 (JAK2-STAT3) pathway in reduction of myocardial ischemia-reperfusion (I/R) injury by sufentanil postconditioning in dogs.Methods Twenty-four healthy dogs of either sex,weighing 10-15 kg,were randomly divided into 4 groups (n =6 each):sham operation group (group S); I/R group; sufentanil postconditioning group (group PO) and sufentanil postconditioning + specific JAK2 inhibitor AG490 group (group AG).In groups I/R,PO and AG,myocardial I/R was produced by occlusion of left anterior descending coronary artery for 30 min followed by 120 min reperfusion.In groups PO and AG,sufentanil 0.6 μg/kg was infused intravenously over 5 min before reperfusion and in addition in group AG,AG490 1 mg/kg was injected intravenously before sufentanil infusion.Myocardial specimens were taken at the end of 120 min reperfusion for microscopic examination and determination of the expression of caspase-3 and p-STAT3 by immuno-histochemistry and myocardial cell apoptosis index (AI) by TUNEL.Results AI and the expression of caspase-3 and p-STAT3 were significantly higher in groups I/R,PO and AG than in group S ( P ＜ 0.05).Compared with group I/R,AI and the expression of caspase-3 were significantly decreased in groups PO and AG,the expression of p-STAT3 was significantly increased in group PO,and the expression of p-STAT3 was significantly decreased in group AG ( P ＜ 0.05).AI and the expression of caspase-3 were significantly higher and the expression of p-STAT3 was significantly lower in group AG than in group PO (P ＜ 0.05).The pathologic changes were significantly attenuated in group PO compared with groups I/R and AG.Conclusion JAK2-STAT3 pathway is involved in reduction of myocardial I/R injury by sufentanil postconditioning in dogs.

ObjectiveTo evaluate the efficacy and safety of spironolactone in treatment NYHA (New York Heart Association) class Ⅰ - Ⅱ heart failure patients. MethodsEighty eight patients with NYHA classⅠ- Ⅱheart failure were randomized to sprionolactone or placebo groups.Patients were assessed by echocardiography and 6-minute walking test (6MWT) ,plasma aldosterone and NT-proBNP were measured before and 6 months after treatment; serum potassium and creatinine were monitored through the study. ResultsThe E/A and E/E' in spironolactone group were significantly lower than those of control group (1.2±0.7 vs. 1.7±0.6, P=0.007;15.2±2.3vs. 17.7±3.0, P=0.000). No differences were observed in left ventricular end-diastolic diameter (LVEDD) and left ventricular ejection fraction (LVEF)between two groups after treatment. Left ventricular mass index (LVMI) in spironolactone group decreased after theatment [(117 ±27) g/m2 vs.(112 ± 19) g/m2,P = 0.044]. Plasma aldosterone level in spironolactone group after treatment was significantly lower than that of control group [ ( 157 ± 16) ng/L vs.( 165 ± 16) ng/L, P =0. 021 ]. Although there were no differences in plasma NT-proBNP level between two groups after treatment, it decreased significantly comparing with that of before treatment ( P = 0. 000). No differences were observed in serum potassium and creatinine between two groups after treatment. However serum potassium and creatinine in spironolactone group increased significantly after treatment [ ( 83 ± 18 )pmol/Lvs. (87 ± 22) μmol/L, P =0.047; (4.4 ±0.4) mmoL/L vs. (4.5 ±0.6) mmol/L, P =0. 012]. ConclusionSpironolactone can alleviate cardiac remolding and diastolic function in NYHA classⅠ - Ⅱ heart failure patients.

Objective: To observe the effects of Tangnaikang (TNK), a compound traditional Chinese herbal medicine, on glucose metabolism and insulin resistance in obese Zucker rats. Methods: Twelve male obese Zucker rats, 6 weeks old, were randomly divided into control group and TNK group (3.24 g/kg) after being fed for 2 weeks. All rats received high-fat diet and 4-week treatment. Body weight and blood glucose were tested every week. Oral glucose tolerance test (OGTT) was performed and fasting insulin level was tested on days 0, 14 and 28. Triglyceride, cholesterol, low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C) and free fatty acids (FFA) were tested on day 28. Glucose infusion rate (GIR) was tested by hyperinsulinemic-euglycemic clamp from day 29. The protein expressions of protein kinase B (Akt), phospho-Akt (p-Akt) (Thr308) and glucose transporter protein 4 (GLUT4) in skeletal muscle and GLUT4 in adipose tissue were measured after hyperinsulinemic-euglycemic clamp test. Results: Compared with the control group, the fed blood glucose level and glucose level of OGTT at 120 min had a significant decline in TNK group on day 28, and TNK caused no alteration of the fasting serum insulin, and the GIR increased significantly in hyperinsulinemic-euglycemic clamp study. Furthermore, TNK increased Akt and p-Akt (Thr308) protein expressions in skeletal muscle and decreased the protein expression of GLUT4 in white adipose tissue. Body weight, and triglyceride, cholesterol, LDL-C and FFA contents were slightly decreased in the TNK group, but there were no statistically significant effects. Conclusion: TNK increases the protein expressions of Akt and p-Akt (Thr308) of the signal transduction pathway to influence the translocation of GLUT4 in skeletal muscle and improves glucose metabolism by reducing insulin resistance.

Objective To investigate the clinical efficacy of umbilical blood stem cell transplantation (UCBSCT) on the treatment of hepatitis B liver cirrhosis. Methods Forty-eight patients with hepatitis B liver cirrhosis were enrolled and divided into the treatment group and the control group. There were 25 patients in the treatment group , who received UCBSCT treatment based on conventional liver protection treatment and 23 patients in the control group , who received conventional liver protection treatment. The changes of liver function , coagulation function, clinical symptoms, signs and side effects were studied before the treatment and at 2, 4, 12 and 24 weeks post-treatment. Results The levels of albumin, cholinesterase, and prothrombin activity in the treatment group were higher than those before treatment and were higher than those in the control group. The parameters in the control group were not significantly changed before and after the treatment (P > 0.05). The levels of alanine aminotransferase,aspartate aminotransferase,total bilirubin in both two groups were not significantly changed before and after the treatment (P > 0.05). After 4-week treatment,the differences on improvement of appetite , lacking in strength , abdominal distension , ascites were statistically significant in the treatment group compared with the control group (P < 0.05). No adverse reactions were observed in all groups. Conclusion UCBSCT on the treatment of hepatitis B liver cirrhosis is safe and reliable.

Objective To explore the relationship between interleukin(IL)-13 RS20541 gene polymorphism and susceptibility of enterovirus 71 (EV71) infection in children's hand-foot-mouth disease.Methods Blood samples were collected from 123 children with EV71 infection from the Central Hospital of Xiangtan (experimental group),and they were divided into mild EV71 infection group (n =62) and severe EV71 infection group (n =61) according to their severity.And 52 healthy children without EV71 infection were selected as the controls,with age and sex matched.Two mL blood samples stored in the-80 ℃ freezer,were collected from all subjects,and DNA was extracted by Beijing ubiolab genetic technology company limited.The SNaPshot was used to determine genotype for G/A polymorphism at RS20541 position of IL-13 gene.SPSS 18.0 software was used to analyze the data.Results IL-13 RS20541 loci had 3 genotypes:AA,GA,GG;the frequency of AA,GA,GG in the experimental group was 4.07％,44.71％,51.22％,which was significantly lower than that in the healthy control group (the frequency of AA,GA,GG were 11.54％,32.69％,55.77％),there was no statistically significant difference in genotypes (x2 =4.676,P ＞ 0.05);there was no statistically significant difference in allele frequency (the frequency of A,G in experimental group was 26.42 ％,73.58 ％,and that of the healthy control group was 27.88 ％,72.12 ％;x2 =0.080,P ＞ 0.05).EV71 infection caused by mild group,severe group and healthy controls genotype frequencies between the 3 groups [(AA + GA) were 53.22％,44.26％,44.23％;GG were 46.78％,55.74％,55.77％;x2 =1.294,P ＞ 0.05] and allele frequency (A were 30.65％,22.13％,27.88％;G were 69.35％,77.87％,72.12％;x2 =2.349,P ＞ 0.05) among the mild group,severe group and healthy control group had no statistical significance.Conclusion There is no correlation between the IL-13RS20541 gene polymorphism and EV 71 infection in children with the hand-foot-mouth disease.

Objective:To analyse the role of Bcl-6/Blimp-1/IL-21 in pathogenesis of primary Sj?gren′s syndrome(PSS),the relationship between Bcl-6/Blimp-1/IL-21 expression and labial gland biopsy grading.Methods:Immunohistochemical(IHC) method was used to detect the expression of Bcl-6/Blimp-1 in salivary gland between 30 cases pations with PSS in disease group and 11 cases patients with mucous cyst,lower lip trauma in control group,and the serum IL-21 levels between disease group of 40 cases patients with PSS and 30 cases healthy volunteers were measured by enzyme-linked immunosorbent assay( ELISA) ,relations among 15 patients with IL-21 expression levels with labial gland biopsy grading.Results:The expression levels of Bcl-6/Blimp-1/IL-21 in disease group were higher than control group;in disease group,with pathological grades increased,the expression levels of Bcl-6/Blimp-1/IL-21 were also raised.Conclusion:①Bcl-6/Blimp-1/IL-21 may participate in the pathogenesis of PSS;Bcl-6/Blimp-1/IL-21 is associated with infiltration lymphocytes of salivary gland.