Abstract:Aim To investigate the effect and possible mechanism of oxymatrine injection (OMT) on cardiac function and ventricular remodeling in doxorubicin-induced chronic heart failure (CHF) rats. Methods The rat model with CHF was established by intraperitoneal injection (ip) of adriamycin (1.5 mg/kg, twice a week for 6 weeks).The model group, captopril injection (CTP, 6.5 mg/kg) group and OMT low (25 mg/kg), medium (50 mg/kg), high (100 mg/kg) dose group were set up, and the control group was set up, with 10 rats in each group. The rats in each group were treated by intraperitoneal injection once a day. 4 weeks later, the cardiac function indexes (left ventricular end-systolic diameter (LVESD), left ventricular end-diastolic diameter (LVEDD), left ventricular fractional shortening (LVFS), stroke volume (SV)) were measured through animal ultrasound; the content of cardiac troponin I (cTnI), brain natriuretic peptide (BNP), stromelysin-2 (ST2) in serum were detected by enzyme-linked immunosorbent assay (ELISA); the left ventricular hypertrophy index (LVHI) was calculated; the morphological changes and fibrosis of myocardial tissue were observed by HE staining and Masson staining; the expression of collagen I (Coll-1), α smooth muscle actin (α-SMA), transforming growth factor-β1 (TGF-β1), Smad2, p-Smad2, Smad3, p-Smad3, Smad7 were detected by Western blot. Results Compared with the model group, the LVESD and LVEDD were decreased in CTP group and OMT low, medium, high dose groups; the LVFS, LVEF and SV were increased (all P＜0.05). The contents of cTnI, BNP and ST2 were decreased (all P＜0.05). The pathological changes and fibrosis of myocardial tissue were significantly improved, and the collagen volume fraction (CVF) were decreased (P＜0.05). The expressions of Coll-1, α-SMA and TGF-β1 were decreased; the ratio of p-Smad2/Smad2 and p-Smad3/Smad3 were decreased; the expression of Smad7 were increased (all P＜0.05). The effect of OMT on various indexes of CHF rats were dose-dependent, and the effect of OMT high-dose group on various indexes were significantly better than those of CTP group. Conclusion OMT can improve cardiac function and inhibit ventricular remodeling in CHF rats, which mechanism may be related to the inhibition of TGF-β1/Smads signaling pathway.

Abstract:Aim To explore the effect of brain-derived neurotrophic factor (BDNF) on Sestrin2 expression and angiogenesis-related mechanisms in endothelial cells. Methods Human umbilical vein endothelial cells (HUVEC) were treated with BDNF (100 μg/L) for 1 h, 2 h, 4 h, 6 h, 8 h, and the protein and mRNA expression of Sestrin2 were detected with immunofluorescent staining, Western blot and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. HUVEC were divided into six groups:control group, BDNF (100 μg/L) group, BDNF+TrkB-Fc (1 mg/L) group, BDNF+KT-5823 (500 nmol/L) group, BDNF+L-NAME(NG-nitro-L-arginine methyl ester) (10－4 mol/L) group, BDNF+DMSO(dimethyl sulfoxide)group; after intervention for 4 h, the expression of Sestrin2 was detected with Western blot. HUVEC were divided into four groups:control group, BDNF (100 μg/L) group, BDNF+Sestrin2 siRNA group, BDNF+control siRNA group; after intervention for 6 h, the capacities of cell migration and tube formation were analysed.Results Sestrin2 mRNA increased in 2 h, 4 h, 6 h group compared with that of the 0 h, 1 h group (P＜0.001), while the protein expression of Sestrin2 increased in 2 h, 4 h, 8 h group compared with that of the 0 h, 1 h group (P＜0.05). BDNF-induced increase in Sestrin2 expression was abolished by L-NAME and PKG inhibitor (P＜0.001). BDNF-induced cell migration and tube formation were completely blocked because of the suppressed expression of Sestrin2 by Sestrin2 siRNA (P＜0.01). Conclusion BDNF confers certain aspects of its proangiogenic capacity through NO/PKG/Sestrin2 pathway.

Abstract:Aim To investigate the expression and clinical significance of Long chain non coding RNA TUG1 (LncRNA TUG1) and microRNA 138-5p(miR-138-5p) in plasma of patients with chronic heart failure(CHF). Methods The expression of LncRNA TUG1 and miR-138-5p in plasma of 148 patients with coronary heart disease complicated with chronic heart failure (CHF group) and 40 healthy persons (control group) were detected by real-time quantitative PCR. The correlation between the expression of TUG1 and miR-138-5p and clinical parameters was analyzed. ROC curves were used to analyze the potential of TUG1 and microRNA-138-5p for early diagnosis of CHF. Kaplan Meier survival analysis was used to analyze the effects of TUG1 and miR-138-5p on the 2-year survival rate of CHF. Results Compared with the control group, the expression of TUG1 and BNP in the plasma of patients with chronic heart failure was significantly higher (P<0.001), while the expression of microRNA-138-5p in the plasma of patients with chronic heart failure was significantly lower (P<0.05). The expression of TUG1 was positively correlated with the expression of BNP in serum of CHF patients (r=0.682, P<0.001), but negatively correlated with the expression of microRNA-138-5p and left ventricular ejection fraction (LVEF) (P<0.05). The area under curve (AUC) of LncRNA TUG1 as a biomarker for the diagnosis of chronic heart failure was 0.868 (95% CI 0.590~0.960, P<0.001). The area under curve (AUC) of microRNA-138-5p as a biomarker for the diagnosis of chronic heart failure was 0.607 (95% CI 0.620~0.940,P<0.001).Kaplan-Meier analysis showed that the 2-year median survival time of patients with high expression of LncRNA TUG1 was shorter than that of patients with low expression, and the difference was statistically significant (χ2=19.77, P<0.001). The median 2-year survival time of patients with high expression of microRNA-138-5p was longer than that of patients with low expression, with significant difference (χ2=11.97, P<0.001). The high expression of TUG1 in plasma of patients with chronic heart failure is related to the diagnosis and prognosis of CHF. Conclusion The high expression of TUG1 in patients with chronic heart failure is negatively correlated with the expression of miR-138-5p, which can be used as a biomarker for early diagnosis and prognosis evaluation of CHF.

Abstract:Aim To explore the characteristics of heart rate variability (HRV) and its short-term prognosis in patients with chronic heart failure complicated with type 2 diabetes mellitus (T2DM). Methods 105 patients with chronic heart failure were selected as heart failure group. Patients were divided into simple heart failure group and heart failure with T2DM group. 40 patients with organic heart disease without heart failure who were hospitalized at the same time were selected as control group. To collect the general data, the results of echocardiography and 24-hour ambulatory electrocardiogram of patients in each group, including the standard deviation of the mean of sinus rhythm normal-to-normal intervals (SDNN), ratio of the total number of sinus R-R intervals to the height of the histogram of sinus R-R intervals (TRI), the square root of the mean of the sum of the squares of differences between adjacent NN intervals (rMSSD) and the proportion of differences between successive R-R intervals that are greater than 50 ms (PNN50). The differences of clinical data between control group and heart failure group, heart failure subgroups were compared. The HRV indexes of different diabetic course in heart failure with T2DM group were analyzed. The patients in heart failure group were followed up for 3 months to compare the number of re-hospitalization due to heart failure. Results The course of disease, the total cost, B-type natriuretic peptide (BNP), left ventricular end diastolic diameter (LVEDd), the slowest heart rate and the average heart rate were higher in the heart failure group than those in the control group, and the hemoglobin (Hb), left ventricular ejection fraction (LVEF), cardiac output (CO), SDNN, TRI, rMSSD and PNN50 were lower than those in the control group. The multivariate logisitic regression analysis showed that the course of disease, BNP and the slowest heart rate were independent risk factors for heart failure in patients with organic heart disease. Total cost, the number of β-blocker used, course of disease, fasting blood glucose, BNP, NYHA class Ⅳ cases, the average heart rate and the fastest heart rate were higher in heart failure with T2DM group than those in simple heart failure group, and SDNN, TRI, rMSSD and PNN50 were lower than those of simple heart failure group (P<0.05). Fasting blood glucose was associated with T2DM in patients with heart failure. In patients with T2DM more than 10 years, the levels of rMSSD and PNN50 decreased significantly (P<0.05). The number of re-hospitalization in heart failure complicated with T2DM group was higher than that in simple heart failure group (P<0.05). Conclusions The heart autonomic nervous function in patients with chronic heart failure is lower than in patients with organic heart disease without heart failure. T2DM mellitus can further decrease the HRV of patients with chronic heart failure, damage the heart autonomic nervous function and affect the short term prognosis of patients with chronic heart failure.

Abstract:Aim To explore the circulating levels of long intergenic non-coding RNA predicting cardiac remodeling (LIPCAR) in patients with chronic heart failure (CHF) and combined renal insufficiency. Methods Venous blood samples were collected from 80 control subjects, 188 patients with CHF, including 80 patients with renal insufficiency, LIPCAR was evaluated by real-time PCR. Results The expression of LIPCAR was significantly different between the control group and the CHF group (1.08±0.28 vs.0.79±0.31, P＜0.01). In addition, the subgroup analysis of CHF showed that the patients complicated with renal insufficiency exhibited a tendency of lower levels of LIPCAR. Furthermore, LIPCAR was negatively correlated with NT-proBNP, whereas positively correlated with ejection fraction (P＜0.01). AUC was 0.777(95%CI＝0.720～0.835) for LIPCAR to judge CHF. Conclusion Circulating LIPCAR levels were significantly downregulated in patients with CHF, especially in those complicated with renal insufficiency. It may be valuable as a potential biomarker for CHF.

Abstract:Aim To analyze the clinical features of patients with chronic heart failure (CHF) complicated with renal insufficiency, and to explore the risk factors of CHF with renal insufficiency. Methods 385 cases of CHF patients who were treated in department of cardiology of our hospital were collected, including 211 males and 174 females, with an average age of 69.62±8.59 years. Applying method of retrospective control study, according to estimated glomerular filtration rate (eGFR), the CHF patients were divided into renal insufficiency group [eGFR＜60 mL/(min·1.73 m2)] and non renal insufficiency group [eGFR≥60 mL/(min·1.73 m2)]. The general information, heart basic diseases, concomitant disease, contrast agent application history, echocardiographic parameters, renal function, blood lipids, N-terminal pro-B-type natriuretic peptide (NT-proBNP), D-dimer, hemoglobin, urine microalbumin and other laboratory examination indexes were analyzed and compared between the two groups. Multivariate Logistic regression was used to analyze the risk factors of CHF patients with renal insufficiency. Results The incidence of renal insufficiency was about 42.1% in 385 patients with CHF. Univariate analysis showed that the differences of age, NYHA heart function classification, hypertension, anemia, diabetes, atrial fibrillation were statistically significant between renal insufficiency group and non renal insufficiency group (P＜0.05). The levels of serum creatinine, urea nitrogen, uric acid, cystatin C and NT-proBNP in renal insufficiency group were higher than those in non renal insufficiency group, and left ventricular ejection fraction and hemoglobin level were lower than those in non renal insufficiency group (P＜0.05). Multivariate Logistic regression analysis showed that age (OR 1.3,5%CI 1.067-2.231), NYHA heart function classification (OR 1.0,5%CI 1.054-3.212), hypertension (OR 1.7,5%CI 1.175-3.292) and cystatin C (OR 1.9,5%CI 1.027-3.851) were independently associated with renal insufficiency. Conclusion The incidence of renal insufficiency was higher in patients with CHF. The elder age, NYHA heart function Ⅳ classification, hypertension and serum cystatin C were independent risk factors of CHF patients with renal insufficiency.

Abstract:Aim To explore the serum levels of interleukin-1 (IL-1) and interleukin-6 (IL-6) in patients with chronic heart failure accompanied with renal damage and its clinical significance. Methods 220 New York Heart Association (NYHA) functional class Ⅱ~Ⅳ patients with chronic heart failure were enrolled in this study. Serum levels of blood urea nitrogen (BUN), serum creatinine (SCr), N-terminal pro-B-type natriuretic peptide(NT-proBNP), IL-1 and IL-6 were detected, microalbuminuria (MAU) was detected randomly, left ventricular ejection fraction (LVEF) was measured by echocardiography, glomerular filtration rate (eGFR) was calculated. The levels of IL-1, IL-6, NT-proBNP, MAU and eGFR in different cardiac function groups were compared and the levels of IL-1 and IL-6 in different renal function groups were compared. The relationship between IL-1, IL-6 and NYHA functional classification, NT-proBNP, LVEF, eGFR and MAU were analysed. Results The serum levels of IL-1, IL-6, NT-proBNP and MAU in NYHAⅢ～Ⅳ groups were higher than NYHA Ⅱgroup (P＜0.05), the eGFR of NYHAⅢ～Ⅳ groups were lower than NYHA Ⅱgroup (P＜0.05), and the levels of IL-1, IL-6, NT-proBNP and MAU in NYHA Ⅳ group were higher than NYHA Ⅲ group (P＜0.05). In NYHA functional class Ⅲ～Ⅳ patients the serum levels of IL-1, IL-6, NT-proBNP and MAU in 30%≤EF≤50% group were higher than EF＞50%group (P＜0.05), the eGFR of 30%≤EF≤50% group were lower than EF＞50% group (P＜0.05), the serum levels of IL-1, IL-6, NT-proBNP and MAU in EF＜30% group were higher than 30%≤EF≤50% group (P＜0.05). In NYHA functional class Ⅲ～Ⅳpatients the serum levels of IL-1, IL-6 in renal dysfunction group [eGFR＜60 mL/(min.1.73m2)] were higher than normal renal function group [eGFR≥60 mL/(min.1.73m2)](P＜0.05). The serum levels of IL-1, IL-6 and eGFR had no obvious difference in different basic etiologies groups (P＞0.05). The serum levels of IL-1 and IL-6 were positively correlated with the NYHA functional classification, MAU and NT-proBNP (P＜0.05), negatively correlated with LVEF and eGFR (P＜0.05). Conclusion The serum levels of IL-1 and IL-6 are elevated in patients with chronic heart failure and correlated with the severity of chronic heart failure and renal damage, and detecting IL-1, IL-6 may provide some assistances for assessing the severity of heart failure in clinical.

Abstract:Aim To analyze the prognostic value of hypoalbuminemia for long-term outcome in elderly patients with chronic heart failure after propensity score stratification. Methods 1271 consecutive elderly patients with chronic heart failure were enrolled and divided into two groups according to serum albumin concentration on admission ＜35 g/L(hypoalbuminemia group) and ≥35 g/L(normoalbuminemia group). Propensity score stratification was conducted to reduce confounding bias. And then COX proportional-hazards regression modeling was used to evaluate the prognostic value of hypoalbuminemia for long-term outcome for the pre-stratification and post-stratification data. Results Before stratification, hypoalbuminemia group was older, had higher NYHA status, direct bilirubin, alkaline phosphatase, creatinine, BNP, less hypertension; lower hemoglobin, indirect bilirubin, TC, serum sodium on admission, EF on admission; faster heart rate on admission. During the median 23.9-month follow-up, there were 165 deaths(13.0%) for all causes, 76 in hypoalbuminemia group(46.1%), 260 in normoalbuminemia group(23.5%). The univariate COX regression analysis indicated that hypoalbuminemia was the strong independent risk for long-term mortality in elderly patients with chronic heart failure(HR was 2.540, 95% CI was 1.966~3.282, P<0.001). Adjusted for propensity score, the multivariate COX regression analysis indicated that the risk of long-term mortality in hypoalbuminemia patients increased 56.1% than the normoalbuminemia patients(HR was 1.561, 95%CI was 1.186~2.054, P＝0.001). After adjustment using propensity score stratification, the overall HR was 1.724, 95%CI was 1.311~2.268, which indicated that the risk of long-term mortality in hypoalbuminemia patients increased 0.724-time than the normoalbuminemia patients. Conclusions Hypoalbuminemia indicated the increasing of long-term mortality in elderly patients with chronic heart failure. It was possible to improve the prognosis of elderly patients with chronic heart failure by correcting hypoalbuminemia.

Abstract:Aim Heat shock protein 47 （HSP47） is a highly conservative proteint. It is considered as the pro-collagen-specific chaperone protein and plays an important function in the process of collagen deposition and fibrosis. The thesis investigates the molecular mechanisms of structural ventricular remodeling in chronic heart failure （CHF）. Methods The serum and right atrial tissues samples were taken from 60 patients with cardiac surgery. They were divided into three groups: 20 samples were in NYHA Ⅰ group, 20 samples were in NYHA Ⅱ group, 20 samples were in NYHA Ⅲ group. The mRNA amounts of HSP47 were studied by real time quantitative polymerase chain reaction （RT-PCR） method, and the HSP47 and procollagen Ⅰ carboxy terminal propeptide （PⅠCP） in serum were measured by enzyme-linked immunosorbent assay （ELISA） method. Results （1）The HSP47 mRNA level significantly increased in both NYHA Ⅱ group and NYHA Ⅲ group compared with NYHA Ⅰ group （P<0.05）. Also the mRNA level of HSP47 significantly increased in the NYHA Ⅲ group compared with NYHA Ⅱ group （P<0.05）. （2）There were conspicuous and independent direct correlation between the expression of HSP47 and the HSP47, PⅠCP levels in serum （r＝0.704, P<0.05 r＝0.811, P<0.05）. Conclusion The HSP47 mRNA level increased in CHF, and it indicates that HSP47 gene may be involved in the ventricular remodeling.

Abstract:Aim To explore the application of carbohydrate antigen 125 （CA125） and amino-terminal pro-brain natriuretic peptide （NT-proBNP） in patients with chronic heart failure. Methods 284 patients with cardiac diseases were divided into four groups （Heart function Ⅰ,Ⅱ,Ⅲ,Ⅳ） according to New York Heart Association （NYHA） class and their serum concentrations of CA125 and NT-proBNP were tested respectively. Then,combined with renal function we respectively compared the level of CA125 and NT-proBNP in the patients with normal renal function or impaired renal function. Results CA125 and NT-proBNP had a significant correlation （r＝0.49,P<0.01） in 284 chosen patients. NT-proBNP concentration was increasing with the level of heart function grade in patients with heart function （Ⅰ,Ⅱ,Ⅲ）（P<0.05）,but NT-proBNP concentration was lower in the patients with heart function Ⅳ than the patients with heart function Ⅲ. However,CA125 concentration was significantly increasing with the level of heart function grade （P<0.05）. In addition combined with renal function,CA125 and NT-proBNP concentration were both significantly increasing with the level of heart function grade （P<0.05） and had a significant correlation （r＝0.700,P<0.01） in 145 patients with normal renal function （eGFR≥60 mL/（min·1.73 m2））. Among 139 patients with impaired renal function （eGFR＜60 mL/（min·1.73 m2））,CA125 and NT-proBNP had a significant correlation （r＝0.292,P<0.01）,NT-proBNP concentration was increasing with the level of heart function grade in patients with heart function （Ⅰ,Ⅱ,Ⅲ） （P<0.05）,but NT-proBNP concentration was significantly lower in the patients with heart function Ⅳ than the patients with heart function Ⅲ （P<0.05）,however CA125 concentration was significantly increasing with the level of heart function grade （P<0.05）. Conclusions Concentration of CA125 and NT-proBNP in serum showed a significant correlation with heart function and could be valuable index in the diagnosis of heart failure. Impaired renal function could interfere with the NT-proBNP concentration,but did not affect the CA125 concentration. Thus,NT-proBNP combined with CA125 would be very useful method to establish diagnosis and treatment in heart failure,especially with renal dysfunction.