Attenuation of Myocardial Fibrosis Using Molecular Hydrogen by Inhibiting the TGF-ß Signaling Pathway in Spontaneous Hypertensive Rats.

BACKGROUND: Previous studies have shown that hydrogen can antagonize the fibrosis of various organs. We investigated whether hydrogen-rich saline (HRS) can attenuate myocardial fibrosis in spontaneously hypertensive rats (SHRs) and clarified the mechanisms involved. METHODS: We examined the effect of HRS and pirfenidone (PFD) on myocardial fibrosis in SHR. Systolic blood pressure, left ventricular mass index (LVMI), and heart weight index (HWI) were measured, Masson trichrome staining was performed. We assessed the role of superoxide dismutase (SOD), malondialdehyde (MDA), Alpha-smooth muscle actin (α-SMA), collagen I, collagen III, and tissue inhibitors of metalloproteinases (TIMPs) in myocardium. We detected the concentrations of procollagen type-I C-terminal propeptide (PICP), procollagen type-III N-terminal propeptide (PIIINP), and angiotensin II (Ang II) in rat serum. Furthermore, the relative protein levels of the transforming growth factor beta (TGF-ß)/Smad pathway were tested. RESULTS: We discovered that HRS decreases LVMI (P < 0.05) and HWI (P < 0.05) in vivo. Compared with model group, HRS decreases the level of collagen volume fraction (P < 0.0001), collagen I (P < 0.001), and collagen III (P < 0.001) in myocardium, and Ang II (P < 0.05), PICP (P < 0.001), and PIIINP (P < 0.05) in serum. In addition, HRS downregulates the expression of MDA (P < 0.01), α-SMA (P < 0.05), and TIMPs (P < 0.05), and increased SOD (P < 0.05). Furthermore, HRS downregulated the expression levels of TGF-ß1 (P < 0.0001), Smad3 (P < 0.0001), and Smad2/3 (P < 0.001), but had no effect on Smad7 expression (P > 0.05). PFD had similar effect compared with HRS and control group. CONCLUSIONS: HRS reduced oxidative stress and improved myocardial collagen content, which may be related to inhibition of the TGF-ß signaling pathway. This suggests that HRS is an effective therapeutic strategy for myocardial fibrosis.

Evaluation of left ventricular systolic function using synchronized analysis of heart sounds and the electrocardiogram.

BACKGROUND: Heart failure is a major health concern and often requires echocardiography to confirm the diagnosis. We introduce a new method that uses a wearable heart sound and electrocardiogram (ECG) device that can be used in the outpatient setting. OBJECTIVE: The purpose of this study was to determine the value of synchronized analysis of heart sounds and ECG in identifying patients with depressed left ventricular ejection fraction (dLVEF) <50%. METHODS: One hundred eighty-nine patients (76 with dLVEF; 113 with normal ejection fraction) were enrolled. All were admitted to the hospital because of dyspnea or chest discomfort. N-Terminal pro-B-type natriuretic peptide (NT-proBNP) was measured in all patients. LVEF was determined by echocardiography. Heart sound and ECG signals were simultaneously recorded using the wearable synchronized phonocardiogram and ECG device. Heart sound and ECG signals were automatically analyzed using wavelet analysis and utilized to determine electromechanical activation time (EMAT), EMAT/RR, S1-S2 time, and S1-S2/RR. RESULTS: EMAT in the dLVEF group was significantly higher than that in the control group (159.82 ± 83 ms vs 91.58 ± 28 ms). Pearson correlation test showed a negative correlation between EMAT and LVEF (r = -0.449; P <.001). Receiver operating characteristic curve analysis demonstrated that the sensitivity and specificity of EMAT ≥104 ms for the diagnosis of EF <50% were 92.1% and 92%, respectively. Patients with intermediate NT-proBNP values were identified as dLVEF by EMAT ≥104 ms, with sensitivity of 93.5% and specificity of 92.8%. CONCLUSION: The heart sound and ECG signal index EMAT contributes to the diagnosis of EF <50% and is especially helpful in patients with an inconclusive NT-proBNP value.

Bioinformatic screening for key miRNAs and genes associated with myocardial infarction.

Despite significant advances in understanding of the causes of and treatment of myocardial infarction (MI) in recent years, morbidity and mortality is still high. The aim of this study was to identify miRNA and genes potentially associated with MI. mRNA and miRNA expression datasets were downloaded from the Gene Expression Omnibus database (http://www.ncbi.nlm.nih.gov/geo/). Interactions between miRNA and the expression and function of target genes were analyzed, and a protein-protein interaction network was constructed. The diagnostic value of identified miRNA and genes was assessed. Quantitative RT-PCR was applied to validate the results of the bioinformatics analysis. MiR-27a, miR-31*, miR-1291, miR-139-5p, miR-204, miR-375, and target genes including CX3CR1,HSPA6, and TPM3 had potential diagnostic value. The genes TFEB,IRS2,GRB2,FASLG,LIMS1,CX3CR1,HSPA6,TPM3,LAT2,CEBPD,AQP9, and MAPKAPK2 were associated with recovery from MI. In conclusion, the identified miRNA and genes might be associated with the pathology of MI.

Evaluation of P wave Dispersion and Tissue Doppler Imaging for Predicting Paroxysmal Atrial Fibrillation in Patients with Hypertension.

BACKGROUND: There are no previous studies dealing with paroxysmal atrial fibrillation (AF) and hypertension using electrocardiogram and tissue doppler imaging (TDI). The aim of this study was to investigate and identify the predictive indicators for paroxysmal AF in hypertensive patients using P wave dispersion (Pd) and TDI. METHODS: Patients with hypertension were enrolled. Patients with paroxysmal AF were classified as the PAF group, and patients without a history of paroxysmal AF were classified as the NAF group. The clinical data, P wave indicators and TDI indicators were collected and compared between the two groups. RESULTS: A total of 120 patients were enrolled into the study with 40 cases in the PAF group and 80 cases in the NAF group. Compared with NAF group, Pd, maximum P wave duration (Pmax), left ventricular end-diastolic dimension (LVEDd) and left atrial dimension (LAD) were significantly longer (P < .05) in the PAF group. PAL, PAI, PAR, LR, LI and IR were significantly longer (P < .05) in the PAF group than in the NAF group. As for ROC analysis, Pd and PAL had the greatest area under the curve. The best diagnostic value of Pd and PAL was 40ms and 78ms, respectively. The combination of Pd ≥40ms with Pmax ≥ 110ms showed higher specificity and positive predictive value but decreased sensitivity and negative predictive value for paroxysmal AF. CONCLUSIONS: The PAF group had significantly longer atrial electromechanical time and higher Pd compared with NAF group. The combination of Pd and TDI may be helpful to predict the onset of paroxysmal AF in patients with hypertension.

Influence of serum adiponectin level and SNP +45 polymorphism of adiponectin gene on myocardial fibrosis.

Adiponectin plays an important role in the development of hypertension, atherosclerosis, and cardiomyocyte hypertrophy, but very little was known about the influence of serum adiponectin or the adiponectin gene polymorphism on myocardial fibrosis. Our study investigates the influence of the SNP +45 polymorphism of the adiponectin gene and serum levels of adiponectin on myocardial fibrosis in patients with essential hypertension. A case-control study was conducted on 165 hypertensive patients and 126 normotensive healthy controls. The genotypes of adiponectin gene polymorphisms were detected by the polymerase chain reaction (PCR) method. Serum concentrations of procollagen were measured by a double antibody sandwich enzyme-linked immunosorbent assay (ELISA) in all subjects. The integrated backscatter score (IBS) was measured in the left ventricular myocardium using echocardiography. The serum levels of adiponectin in hypertensive patients were significantly lower than those in the normal control group ((2.69±1.0) µg/ml vs. (4.21±2.89) µg/ml, respectively, P<0.001). The serum levels of type-I procollagen carboxyl end peptide (PICP) and type-III procollagen ammonia cardinal extremity peptide (PIIINP) in the hypertension group were significantly higher than those in the control group. In the hypertension group, serum levels of adiponectin were significantly and negatively related to the average acoustic intensity and corrected acoustic intensity of the myocardium (r=0.46 and 0.61, respectively, P<0.05 for both). The serum levels of PICP and PIIINP were significantly different among the three genotypes of SNP +45 (P<0.01). Logistic regression analyses showed that sex and genotype (GG+GT) were the major risk factors of myocardial fibrosis in hypertensive patients (OR=5.343 and 3.278, respectively, P<0.05). These data suggest that lower levels of adiponectin and SNP +45 polymorphism of the adiponectin gene are likely to play an important role in myocardial fibrosis in hypertensive patients.

Predictive value of left ventricular remodeling by area strain based on three-dimensional wall-motion tracking after PCI in patients with recent NSTEMI.

We aimed to explore whether a novel left ventricular performance index, area strain (AS), based on three-dimensional wall-motion tracking (3-D-WMT) done before and after percutaneous coronary intervention (PCI) could predict left ventricular (LV) remodeling in patients with recent non-ST elevation myocardial infarction (NSTEMI). Sixty-one patients (53.6 ± 8.8 years) with recent NSTEMI were enrolled. Coronary angiography and PCI were undertaken for reperfusion. Parameters of myocardial deformation (including LV end-diastolic volume, LV end-systolic volume, LV ejection fraction, LV global and regional peak area strain) were measured by 3-D-WMT before and 1 week after reperfusion therapy. Six months after reperfusion, LV negative remodeling was defined as lack of improvement in LV function, with increase in LV end-diastolic volume ≥15%. Patients were subdivided into remodeled group (n = 25) and non-remodeled group (n = 36) at follow-up. Patients with negative LV remodeling had significantly higher cardiac troponin I (cTnI) levels at baseline (21.21 ± 12.22 vs. 15.56 ± 8.91 ng/mL; p = 0.0357), higher B-type natriuretic peptide (BNP) level (247.56 ± 177.39 vs. 170.53 ± 97.89 pg/mL; p = 0.0336) and reduced global AS (-27.9 ± 4.6% vs. -31.9 ± 4.3%; p = 0.001) than those without remodeling. Global AS at baseline had a significantly close correlation with cTnI level 36 h after MI (r = 0.71, p < 0.001). Moreover, a weak relationship was found between LV global AS at baseline and BNP level 24 h after myocardial infarction (r = 0.423, p < 0.001). By multivariate logistic regression analysis, lack of improvement of global AS 1 week after PCI was found to be a powerful independent predictor of negative LV remodeling at follow-up (OR = 1.41, 95% CI 1.28-3.27, p = 0.003). In particular, a global AS ≤32% (absolute value) showed a sensitivity and a specificity of 86.1% and 68.0% in predicting negative LV remodeling. These data suggest that AS could be used to assess myocardial global and regional LV function with good feasibility and repeatability. Global AS 1 week after PCI is a good independent predictor of negative LV remodeling after 6-month follow-up.

Value of left ventricular regional ejection fraction determined by real-time three-dimensional echocardiography in diagnosis of aneurysm: compared with left ventriculography.

BACKGROUND: Regional ejection fraction (EF(R)) measured by real-time three-dimensional echocardiography (RT-3DE) provides a novel method for quantifying left-ventricular (LV) regional systolic function. We aimed to explore the diagnostic value of regional ejection fraction (EFR) derived from RT-3DE in detecting LV aneurysms in patients with myocardial infarction. METHODS: Thirty-eight patients with myocardial infarction were prospectively enrolled and underwent electrocardiography (ECG), two-dimensional echocardiography (2-DE), RT-3DE and left ventriculography (LVG). Subjects with a negative EFR in at least one segment on RT-3DE were considered as having a ventricular aneurysm. We compared the sensitivity, specificity, Youden's index, and positive and negative predictive values of ECG, 2-DE and RT-3DE in determining LV aneurysm with detection by LVG. RESULTS: On LVG an LV aneurysm was diagnosed in 16 (42.1%) patients. The sensitivity and specificity were 62.5% and 86.4% for ECG, 81.2% and 95.4% for 2-DE, and 100.0% and 90.9% for RT-3DE in diagnosing LV aneurysm. Youden's indexes for ECG, 2-DE and RT-3DE were 0.49, 0.77 and 0.91, respectively. Positive and negative predictive values were 76.9% and 76.0% for ECG, 92.9% and 87.5% for 2-DE, and 88.9% and 100.0% for RT-3DE. CONCLUSIONS: RT-3DE-derived EFR provides a novel, reliable index in the diagnosis of LV aneurysm and has excellent sensitivity and specificity.

Quantification of regional volume and systolic function of the left ventricle by real-time three-dimensional echocardiography.

Real-time three-dimensional (3D) echocardiography (RT-3DE) provides a unique technique to evaluate left ventricular regional function in a 3D format. We aimed to explore whether the left ventricular segmental volume and systolic function is uniform and to establish normal values of volume and systolic function parameters of 16 regions in healthy subjects. RT-3DE was performed in 41 normal subjects and four-dimensional (4D)-left ventricle (LV) analysis software and a TomTec workstation were used to analyze data for regional end-diastolic volume (EDV(R)), regional end-systolic volume (ESV(R)), regional stroke volume (SV(R)), regional ejection fraction (EF(R)), ratio of SV(R) to global SV (SV(R/G)) and ratio of SV(R) to global EDV (EF(R/G)). All regional volume and systolic function parameters were not uniform among the left ventricular walls. They all increased in the order of inferior, posterior, lateral, septal, anterior and antero-septal walls with an increasing trend from the apical, middle to basal segments. The systolic function (EF(R), SV(R/G) and EF(R/G)) of the anterior and antero-septal walls was significantly higher than that of the lateral, inferior and posterior walls. And the intra- and interobserver variability for EDV(R), ESV(R), SV(R/G) and EF(R/G) ranged from 2.9% to 5.8%. In conclusion, the regional volume and systolic function of the left ventricle is not uniform and, therefore, a normal left ventricle cannot be regarded as a symmetric model for assessing the regional systolic function. This information may improve the accuracy of RT-3DE techniques in the assessment of the left ventricular regional function. (E-mail: zhangyun@sdu.edu.cn and yaogh@yahoo.com).

[Observation on therapeutic effects of red-hot needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation].

OBJECTIVE: To compare therapeutic effects of red-hot needle therapy and filiform needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation. METHODS: Six hundred cases were randomly divided into a treatment group and a control group, 300 cases in each group. The treatment group were treated with red-hot needle pricking at the proliferative parts and Ashi points as main, and the control group with filiform needle therapy. RESULTS: In the treatment group, 240 cases were cured, 58 cases improved and 2 cases were not cured with an effective rate of 99.3%; and in the control group, 113 cases were cured, 165 cases improved and 22 were not cured with an effective rate of 92.7%, with a significant difference between the two groups (P = 0.001). CONCLUSION: The therapeutic effect of red-hot needle therapy is better than that of filiform needle therapy on nodules of breast of the type of stagnation of liver-qi and phlegm coagulation.

[Effects of losartan, ramipril and their combination on left ventricular remodeling and function in spontaneous hypertensive rats].

OBJECTIVE: To evaluate the effects of losartan, an angiotensin receptor blocker, ramipril, an angiotensin converting enzyme inhibitor, and their combination on left ventricular remodeling and diastolic function in SHR at the same level of blood pressure. METHODS: Sixty-two SHRs and 20 WKYs were divided randomly into 5 groups: WKY-control group, SHR-control group, SHR-ramipril group, SHR-losartan group, and SHR-combination group. Twelve weeks after feeding, 6 rats from each group were randomly selected to undergo hemodynamic examination and then killed to undergo further examinations, and 24 weeks after the remaining rats underwent the same examinations. The hemodynamic examination included the systolic blood pressure (SBP) of the caudal artery, left ventricular systolic pressure (LVSP), left ventricle end diastolic pressure (LVEDP), maximum uprising velocity of left ventricle pressure (dP/dtmax), and maximum declining velocity of left ventricle pressure (-dP/dtmax), and tau. Then the hearts were taken out to measure the weight of heart, undergo pathological examination, measure the intracellular free calcium concentrations, hydroxyproline concentration, interstitial collagen volume fraction (CVF), perivascular collagen area/luminal area (PVCA/LA), the mRNA expression of SR Ca(2+)-ATPase, phospholamban and L-type calcium channel, and the protein levels of SR Ca(2+)-ATPase. The myocardial ultrastructure was analyzed by electron microscopy. RESULTS: The speed, extent, and sustained time of blood pressure decrease were better in the combination group than in the other 2 treatment groups. Twelve and 24 weeks after treatment the levels of LVM/BW in the combination group were significantly lower than those of the control group, however, without significant differences among the 3 treatment groups. The values of LVSP, LVEDP, and tau 12 and 24 weeks after treatment in the 3 treatment groups were all significantly lower and the levels of -dP/dtmax significantly higher than those of the control group (all P < 0.01). The values of CVF in the myocardium and PCVA/VA of the heart wall arteriole 12 and 24 weeks after in the 3 treatment groups were significantly lower than those of the control group, and the CVF in the myocardium 12 weeks after in the combination group was significantly than that of the ramipril group. Microscopy showed that the degree of myocardial fibrosis in the 3 treatment groups were significantly milder than those of the control group, and the ultrastructure improvement improved along with the lapse of time in the sequence of combination group > losartan group > ramipril group. The concentrations of hydroxyproline in cardiac muscle cells of the 3 treatment 12 and 24 weeks after were significantly than those of the control group and decreased gradually time-dependently. The expression of Ca(2+)-ATPase mRNA 24 weeks after of the ramipril, losartan, and combination groups were 53.5%, 72.9%, and 76.7% higher than that of the control group, and the Ca(2+)-ATPase protein expression of the 3 treatment groups were 28.9%, 33.3%, and 49.3% higher than that of the control group. The expression of L-type Ca(2+) channel mRNA of the 3 treatment groups were 51.8%, 76.8%, and 98.2% than that of the control group. CONCLUSION: Both losartan and ramipril reverse LVH and left ventricular diastolic dysfunction. A combination of these two drugs is more effective than single drug treatment for improvement of myocardial fibrosis and ultrastructure. All three-treatment groups can raise calcium-handling proteins mRNA and protein expressions, which may be the underlying molecular mechanisms for their therapeutic effects.

[Relationship between regression of hypertensive left ventricular hypertrophy and improvement of coronary flow reserve].

OBJECTIVE: To elucidate the relationship between regression of hypertensive left ventricular hypertrophy (LVH) and changes of coronary flow reserve (CFR). METHODS: Ninety-six essential hypertensive patients with LVH were randomly divided into 3 groups: ramipril group, losartan group, and combination group. Before the treatment and 6 months after the treatment, left ventricular mass (LVM) was calculated by three-dimensional echocardiography and CFR was evaluated by transesophageal echocardiography with left anterior descending artery. CFR was calculated as the ratio of coronary flow velocity, after intravenous injection of dipyridamole (D), to rest peak velocity (R). All the indexes of the CFR were corrected by LVM. RESULTS: (1) The systolic blood pressure (SBP), diastolic blood pressure (DBP) and LVM were significantly decreased in ramipril group, losartan group and combination group after 6 months treatment (all P < 0.01). Diastolic flow velocity integral was used as one representative index of CFR. Compared with baseline values, the diastolic flow velocity intergral corrected by LVM (D/R DVi(C)) was significantly increased in ramipril group, losartan group and combination group (1.83 +/- 0.61 vs 1.57 +/- 0.58, P < 0.05; 1.94 +/- 0.45 vs 1.53 +/- 0.64, P < 0.01; 2.03 +/- 0.38 vs 1.49 +/- 0.34, P < 0.01). (3) The changes of D/R DVi(C) showed a positive correlation with the changes of LVM (r = 0.579, P < 0.05) and no significant correlation with the decrease of blood pressure (r = 0.288 and 0.295,both P > 0.05). CONCLUSION: Ramipril, losartan and combination of these two drugs all reduce blood pressure and LVH, and increases CFR. CFR corrected by LVM may help assess drug's effect on CFR. Improvement of CFR is associated with the regression of hypertensive LVH.