Detection of an anti-angina therapeutic module in the effective population treated by a multi-target drug Danhong injection: a randomized trial.

It's a challenge for detecting the therapeutic targets of a polypharmacological drug from variations in the responsed networks in the differentiated populations with complex diseases, as stable coronary heart disease. Here, in an adaptive, 31-center, randomized, double-blind trial involving 920 patients with moderate symptomatic stable angina treated by 14-day Danhong injection(DHI), a kind of polypharmacological drug with high quality control, or placebo (0.9% saline), with 76-day following-up, we firstly confirmed that DHI could increase the proportion of patients with clinically significant changes on angina-frequency assessed by Seattle Angina Questionnaire (ΔSAQ-AF ≥ 20) (12.78% at Day 30, 95% confidence interval [CI] 5.86-19.71%, P = 0.0003, 13.82% at Day 60, 95% CI 6.82-20.82%, P = 0.0001 and 8.95% at Day 90, 95% CI 2.06-15.85%, P = 0.01). We also found that there were no significant differences in new-onset major vascular events (P = 0.8502) and serious adverse events (P = 0.9105) between DHI and placebo. After performing the RNA sequencing in 62 selected patients, we developed a systemic modular approach to identify differentially expressed modules (DEMs) of DHI with the Zsummary value less than 0 compared with the control group, calculated by weighted gene co-expression network analysis (WGCNA), and sketched out the basic framework on a modular map with 25 functional modules targeted by DHI. Furthermore, the effective therapeutic module (ETM), defined as the highest correlation value with the phenotype alteration (ΔSAQ-AF, the change in SAQ-AF at Day 30 from baseline) calculated by WGCNA, was identified in the population with the best effect (ΔSAQ-AF ≥ 40), which is related to anticoagulation and regulation of cholesterol metabolism. We assessed the modular flexibility of this ETM using the global topological D value based on Euclidean distance, which is correlated with phenotype alteration (r2: 0.8204, P = 0.019) by linear regression. Our study identified the anti-angina therapeutic module in the effective population treated by the multi-target drug. Modular methods facilitate the discovery of network pharmacological mechanisms and the advancement of precision medicine. (ClinicalTrials.gov identifier: NCT01681316).

TBX20 loss-of-function mutation associated with familial dilated cardiomyopathy.

BACKGROUND: Dilated cardiomyopathy (DCM) is a major cause of congestive heart failure, sudden cardiac death and cardiac transplantation. Aggregating evidence highlights the genetic origin of DCM. However, DCM is a genetically heterogeneous disorder, and the genetic components underlying DCM in most cases remain unknown. METHODS: The coding regions and splicing junction sites of the TBX20 gene were sequenced in 120 unrelated patients with idiopathic DCM. The available close relatives of the index patient carrying an identified mutation and 300 unrelated ethnically matched healthy individuals used as controls were genotyped for TBX20. The functional characteristics of the mutant TBX20 were assayed in contrast to its wild-type counterpart by using a dual-luciferase reporter assay system. RESULTS: A novel heterozygous TBX20 mutation, p.F256I, was identified in a family with DCM transmitted in an autosomal dominant fashion, which co-segregated with DCM in the family with complete penetrance. The missense mutation was absent in 600 control chromosomes and the altered amino acid was completely conserved evolutionarily among various species. Functional assays revealed that the mutant TBX20 had significantly diminished transcriptional activity. Furthermore, the mutation markedly reduced the synergistic activation of TBX20 with NKX2-5 or GATA4. CONCLUSIONS: This study links TBX20 loss-of-function mutation to idiopathic DCM in humans for the first time, providing novel insight into the molecular mechanism underpinning DCM.

Expression characteristics of neutrophil and mononuclear-phagocyte related genes mRNA in the stable angina pectoris and acute myocardial infarction stages of coronary artery disease.

OBJECTIVE: To investigate expression differences of neutrophil and mononuclear phagocyte related gene mRNAs among acute myocardial infarction (AMI), stable angina (SA) and control groups, and then discuss their expression characteristics in the stable angina pectoris (SAP) and AMI stages of coronary artery disease (CAD). METHODS: Whole Human Genome Oligo Microarrays were applied to assess the differential expression characteristics of neutrophil and mononuclear phagocyte related mRNAs in patients with AMI (n = 20), SA (n = 20) and controls (n = 20). RESULTS: (1) Almost all colony-stimulating factors (CSF) and their receptors related mRNAs was up-regulated in AMI and SA groups compared with the control group, and the expression of granulocyte-macrophage colony stimulating factor receptor (GM-CSFR) and granulocyte colony stimulating factor receptor (G-CSFR) mRNAs in the AMI group was significantly up-regulated compared with the other two groups (P < 0.01). (2) The expression of mRNAs related to monocyte chemoattractant protein-1 (MCP-1), CCR2 (MCP-1 receptor) and CXCR2 (IL-8 receptor) was significantly up-regulated (P < 0.01) in AMI group compared with SA and control groups. IL-8 mRNA expression in the AMI group was clearly higher than the controls (P < 0.05). (3) All mRNAs expression related to opsonic receptors (IgG FcR and C3bR/C4bR) was significantly up-regulated in AMI group compared with SA and control group (P < 0.01), and the SA group showed an upward trend compared with controls. (4) Most pattern recognition receptor (PRR)-related mRNAs expression was up-regulated in AMI group compared with SA and control groups. Most toll-like receptor (TLR) mRNAs expression was significantly up-regulated (P < 0.01) than the SA and control groups; macrophage scavenger receptor (MSR) mRNA was significantly up-regulated in AMI group compared with the control group (P < 0.01), and the SA group showed an upward trend compared with the controls. CONCLUSIONS: The expression of most neutrophil and mononuclear-macrophage function related genes mRNAs was significantly up-regulated by stages during the progression of CAD, suggesting that the adhesive, chemotactic and phagocytic functions of neutrophil and mononuclear-macrophage were strengthened in the occurrence and development of coronary atherosclerosis and AMI. This also showed a stepped upward trend as the disease progressed.

Effects of antiarrhythmic peptide 10 on acute ventricular arrhythmia.

OBJECTIVE: To observe the effects antiarrhythmic peptide 10 (AAP10) aon acute ventricular arrhythmia and the phosphorylation state of ischemic myocardium connexin. METHODS: Acute total ischemia and partial ischemia models were established by ceasing perfusion and ligating the left anterior descending coronary artery in SD rats. The effects of AAP10 (1 mg/L) on the incidence rate of ischemia-induced ventricular arrhythmia were observed. The ischemic myocardium was sampled to detect total-Cx43 and NP-Cx43 by immunofluorescent staining and western blotting. the total-Cx43 expression was detected through image analysis system by semi-quantitative analysis. RESULTS: AAP10 could significantly decrease the incidence of ischemia-induced ventricular tachycardia and ventricular fibrillation. During ischemic stage, total ischemia (TI) and AAP10 total ischemia (ATI) groups were compared with partial ischemia (PI) and AAP10 partial ischemia (API) groups. The rates of incidence for arrhythmia in the ATI and API groups (10% and 0%) were lower than those in the TI and PI groups (60% and 45%). The difference between the two groups was statistically significant (P=0.019, P=0.020). The semi-quantitative analysis results of the ischemic myocardium showed that the total-Cx43 protein expression distribution areas for TI, ATI, PI and API groups were significantly decreased compared with the control group. On the other hand, the NP-Cx43 distribution areas of TI, ATI, PI and API groups were significantly increased compared with the control group (P>0.05). AAP10 could increase the total-Cx43 expression in the ischemic area and decrease the NP-Cx43 expression. Western blot results were consistent with the results of immunofluorescence staining. CONCLUSIONS: AAP10 can significantly decrease the rate of incidence of acute ischemia-induced ventricular tachycardia and ventricular fibrillation. Acute ischemic ventricular arrhythmias may have a relationship with the decreased phosphorylation of Cx43 induced by ischemia. AAP10 may stimulate the phosphorylation of Cx43 by increasing the total-Cx43 expression and decreasing the NP-Cx43 expression in the ischemic area, so as to decrease ventricular arrhythmia.

Mutational spectrum of the NKX2-5 gene in patients with lone atrial fibrillation.

Atrial fibrillation (AF) is the most common form of sustained cardiac arrhythmia in humans and is responsible for substantial morbidity and mortality worldwide. Emerging evidence indicates that abnormal cardiovascular development is involved in the pathogenesis of AF. In this study, the coding exons and splice sites of the NKX2-5 gene, which encodes a homeodomain-containing transcription factor essential for cardiovascular genesis, were sequenced in 146 unrelated patients with lone AF as well as the available relatives of the mutation carriers. A total of 700 unrelated ethnically matched healthy individuals used as controls were genotyped. The disease-causing potential of the identified NKX2-5 variations was predicted by MutationTaster and PolyPhen-2. The functional characteristics of the mutant NKX2-5 proteins were analyzed using a dual-luciferase reporter assay system. As a result, two heterozygous NKX2-5 mutations, including a previously reported p.E21Q and a novel p.T180A mutation, were identified in two families with AF transmitted in an autosomal dominant pattern. The mutations co-segregated with AF in the families with complete penetrance. The detected substitutions, which altered the amino acids highly conserved evolutionarily across species, were absent in 700 control individuals and were both predicted to be causative. Functional analyses demonstrated that the NKX2-5 mutants were associated with significantly decreased transcriptional activity compared with their wild-type counterpart. The findings expand the spectrum of NKX2-5 mutations linked to AF and provide additional evidence that dysfunctional NKX2-5 may confer vulnerability to AF, suggesting the potential benefit for the early prophylaxis and personalized treatment of AF.

A novel GATA4 loss-of-function mutation responsible for familial dilated cardiomyopathy.

Dilated cardiomyopathy (DCM) is the most common form of primary myocardial disorder and is associated with substantial morbidity and mortality. Increasing evidence suggests that genetic risk factors play an important role in the pathogenesis of idiopathic DCM. However, DCM is a genetically heterogeneous disease, and the genetic defects responsible for DCM in an overwhelming majority of cases remain to be identified. In the present study, the entire coding region and the splice junction sites of the GATA4 gene, which encodes a cardiac transcription factor essential for cardiogenesis, were sequenced in 150 unrelated patients with idiopathic DCM. The available relatives of the index patient harboring an identified mutation and 200 unrelated ethnically matched healthy individuals used as controls were genotyped. The functional characteristics of the mutant GATA4 were delineated in contrast to its wild-type counterpart using a luciferase reporter assay system. As a result, a novel heterozygous GATA4 mutation, p.V291L, was identified in a family with DCM inherited in an autosomal dominant pattern, which co-segregated with DCM in the family with complete penetrance. The missense mutation was absent in 400 control chromosomes, and the altered amino acid was completely conserved evolutionarily among species. Functional analysis revealed that the GATA4 mutant was associated with significantly diminished transcriptional activity. The findings expand the mutational spectrum of GATA4 linked to DCM and provide novel insight into the molecular etiology involved in DCM, suggesting the potential implications in the early prophylaxis and allele-specific treatment for this common type of cardiomyopathy.

Quantitative evaluation of cardiopulmonary functional reserve in treated patients with pulmonary embolism.

BACKGROUND: There is no research, either at home or abroad, focusing on assessing the cardiopulmonary functional reserve and exercise tolerance in patients with pulmonary embolism (PE), but the benefits of early exercise are well recognized. The goals of this study were to assess cardiopulmonary functional reserve in treated PE patients using the inert gas rebreathing method of the cardiopulmonary exercise test (CPET), and to compare it with traditional methods. METHODS: CPET on the bicycle ergometer were performed in 40 patients with age, gender, body mass index, systolic blood pressure, and pulmonary function matched. The first group was the PE group composed of 16 PE patients (5 male, 11 female) who were given the standard antithrombotic therapy for two weeks. The second group was composed of 24 normal individuals (10 male, 14 female). Both groups were evaluated by cardiac ultrasound examination, 6-minute walking test (6MWT), and CPET. RESULTS: (1) Right ventricular systolic pressure (RVSP) in the PE group increased significantly compared to the control group, (34.81 ± 8.15) mmHg to (19.75 ± 3.47) mmHg (P < 0.01). But neither right atrial end-systolic diameter (RASD) nor right ventricular end-diastolic diameter (RVDD) in the PE patients had changed when compared with the controls. The 6-minute walk distance was significantly reduced in the PE patients compared with normal subjects, (447.81 ± 79.20) m vs. (513.75 ± 31.45) m (P < 0.01). Both anaerobic threshold oxygen consumption (VO(2)AT) and peak oxygen consumption (VO(2)peak) were significantly lower in patients with PE, while CO(2) equivalent ventilation (VE/VCO(2) slope) was higher; VO(2)AT (9.44 ± 3.82) ml×kg(-1)×min(-1) vs. (14.62 ± 2.93) ml×kg(-1)×min(-1) (P < 0.01) and VO2peak (12.26 ± 4.06) ml×kg(-1)×min(-1) vs. (23.46 ± 6.15) ml×kg(-1)×min(-1) (P < 0.01) and VE/VCO(2) slope 35.47 ± 6.66 vs. 26.94 ± 3.16 (P < 0.01). There was no significant difference in resting cardiac output (CO) between the PE and normal groups, whereas peak cardiac output (peak CO) and the difference between exercise and resting cardiac output (ΔCO) were both significantly reduced in the PE group; peak CO (5.97 ± 2.25) L/min to (8.50 ± 3.13) L/min (P < 0.01), ΔCO (1.29 ± 1.59) L/min to (3.97 ± 2.02) L/min (P < 0.01). (2) The 6-minute walk distance did not correlated with CPET except for the VO2 peak in patients with PE, r = 0.675 (P < 0.01). CONCLUSIONS: The cardiopulmonary functional reserve was reduced in patients with PE. CPET is an accurate, quantitative evaluation of cardiopulmonary functional reserve for PE patients.

[The effects of aerobic exercise on cardiac output during exercise in patients with chronic heart failure].

OBJECTIVE: To observe the effects of aerobic exercise on cardiac output during exercise in patients with chronic heart failure (CHF). METHODS: A total of 50 CHF patients (echocardiography measured left ventricular ejection fraction < 0.49) were enrolled in the study and randomly divided into aerobic exercise group (n = 25) and control group (n = 25). Cardiopulmonary exercise testing (CPET) was performed. Patients of aerobic exercise group underwent aerobic exercise according to aerobic exercise prescription and exercise intensity is decided by anaerobic threshold before 10 J/s (1 minute before) of the oxygen consumption. After 6 supervised aerobic exercise training sessions in the hospital, patients were asked to perform the home-based aerobic exercise training. Patients in control group were required to maintain daily physical activities. CPET were reviewed 3 months later. RESULTS: Cardiac output (CO), peak CO, peak cardiac power output (peak CPO), resting heart rate (HR), heart rate at AT (HRAT), HR peak, resting mean arterial pressure (MAP), peak MAP at baseline were similar between aerobic exercise group and control [(4.2 ± 2.0) L/min vs. (3.3 ± 1.0) L/min, (6.2 ± 2.7) L/min vs. (5.2 ± 1.8) L/min, (1.8 ± 2.9) L/min vs. (2.0 ± 1.8) L/min, (1.3 ± 0.5) J/s vs. (1.2 ± 0.5) J/s, (76.8 ± 13.5) beats/min vs. (73.4 ± 11.9) beats/min, (91.5 ± 11.3) beats/min vs. (92.6 ± 12.4) beats/min, (106.0 ± 12.9) beats/min vs. (108.3 ± 17.4) beats/min, (80.8 ± 9.9) mm Hg (1 mm Hg = 0.133 kPa) vs. (87.6 ± 13.3) mm Hg, (98.8 ± 12.4) mm Hg vs. (102.7 ± 13.9) mm Hg, all P > 0.05]. Compared to baseline, CO, peak CO, peak CPO, HR, HRAT, HR peak, MAP, peak MAP after 3 months were similar between aerobic exercise group and control (all P > 0.05). The differences between baseline and 3 months later expressed as ΔCO, Δpeak CO, Δpeak CPO, ΔHR, ΔHRAT, ΔHR peak, ΔMAP, Δpeak MAP were also similar between aerobic exercise group and control group [(-0.7 ± 2.4) L/min vs. (0.7 ± 2.0) L/min, (1.1 ± 2.6) L/min vs. (1.4 ± 2.1) L/min, (0.1 ± 3.7) L/min vs. (-0.2 ± 2.5) L/min, (0.2 ± 1.0) J/s vs. (0.2 ± 0.5) J/s, (-0.4 ± 7.6) beats/min vs. (1.9 ± 9.9) beats/min, (3.4 ± 11.3) beats/min vs. (-2.8 ± 7.6) beats/min, (8.9 ± 14.5) beats/min vs. (3.7 ± 14.4) beats/min, (1.5 ± 12.8) mm Hg vs. (-1.3 ± 11.1) mm Hg, (6.4 ± 18.9) mm Hg vs. (1.3 ± 12.3) mm Hg, all P > 0.05]. CONCLUSION: Three months aerobic exercise training did not improve cardiac output and related parameters during exercise in this cohort patients with CHF.

[Effects of exercise therapy at the intensity of anaerobic threshold for exercise tolerance in patients with chronic stable coronary artery disease].

OBJECTIVE: To investigate the effects of exercise therapy at the intensity of anaerobic threshold (AT) for exercise tolerance in patients with chronic stable coronary artery disease. METHODS: Forty-three patients with chronic stable coronary artery disease (3 patients after coronary arterial bypass graft (CABG) surgery, 22 patients with old myocardial infarction and 18 unstable angina pectoris undergoing successful percutaneous coronary intervention (PCI) finished twice cardiopulmonary exercise test (CPET) and followed their rehabilitation program for 3 months. Thirty-two patients finished their aerobic exercise therapy based on their individual anaerobic thresholds while 11 patients had no exercise therapy. RESULTS: The heart rate at AT intensity (97 ± 9/min) was lower than their traditional minimal target heart rate (112 ± 7/min) and lower than heart rate (115 ± 11/min) at ischemic threshold post-CPET. The O(2) consumption (10.7 ± 2.4 to 12.6 ± 2.9 ml×min(-1)×kg(-1)) (P = 0.04) and workload (37 ± 18 to 47 ± 13 J/s) (P = 0.04) at AT level and the O(2) consumption (15.3 ± 3.1 to 20.6 ± 4.2 ml×min(-1)×kg(-1), P = 0.02) and workload(68 ± 12 and 87 ± 14 J/s, P = 0.01) at peak level markedly increased after 3 months in the exercise group. And the O(2) consumption (15.3 ± 2.9 to 16.2 ± 3.1 ml×min(-1)×kg(-1)) and workload (65 ± 13 to 73 ± 16 J/s) at peak level mild increased after 3 months in the non-exercise group, but their O(2) consumption (11.0 ± 2.7 to 11.3 ± 2.8 ml×min(-1)×kg(-1)) and workload (38 ± 11 to 37 ± 9 J/s) at AT level had no obvious change. CONCLUSION: AT exercise intensity was lower than ischemic threshold post-CPET. Exercise therapy at the intensity of anaerobic threshold can improve oxygen capacity and exercise tolerance.

[Effects of aerobic exercise on exercise tolerance in patients with chronic heart failure].

OBJECTIVE: To explore the effects of aerobic exercise on exercise tolerance in patients with chronic heart failure (CHF). METHODS: A total of 50 CHF patients with left ventricular ejection fraction (LVEF) < 49% by echocardiography were enrolled. And they were randomly divided into exercise group (n = 25) and non-exercise group (n = 25). Cardiopulmonary exercise testing (CPET) was performed. The patients of exercise group underwent an aerobic exercise program in which exercise intensity was decided by anaerobic threshold (AT) before 10 J/s while those of non-exercise group performed daily activities. After 6 sessions of supervised aerobic exercise, the home-based aerobic exercise training began. CPET was re-examined 3 months later. RESULTS: The VO(2) AT, VO(2) peak, Load AT, Load peak, peak VO(2)/HR and VE/VCO(2) slope at baseline were similar between exercise group and non-exercise group (P > 0.05). The VO(2) AT, VO(2) peak, Load AT, Load peak and peak VO(2)/HR in patients of exercise group were increased compared with baseline, The differences between baseline and 3 months later expressed as ΔVO(2) AT, ΔVO(2) peak, ΔLoad AT, ΔLoad peak, Δpeak VO(2)/HR and ΔVE/VCO(2) slope, The differences of ΔVO(2) AT, ΔVO(2) peak, ΔLoad AT, ΔLoad peak and Δpeak VO(2)/HR between two groups were statistically significant [ΔVO(2) AT: 2.8 (1.2 - 3.5) ml×kg(-1)×min(-1) vs -0.3 (-2.8 - 0.1) ml×kg(-1)×min(-1), P < 0.01; ΔVO(2) peak: 3.4 (1.8 - 4.6) ml×kg(-1)×min(-1) vs -0.5 (-1.4 - 0.3) ml×kg(-1)×min(-1), P < 0.01; ΔLoad AT:15.0 (2.5 - 22.5) J/s vs 0.5(-4.2 - 3.8) J/s, P < 0.01; ΔLoad peak: 15.0 (1.3 - 25.0) J/s vs 0.0 (-8.8 - 15.0) J/s, P < 0.05; Δpeak VO(2)/HR: 2.3 (0.0 - 4.0) ml×kg(-1)×beat(-1) vs -0.1 (-0.7 - 1.2) ml×kg(-1)×beat(-1), P < 0.01]. The difference of ΔVE/VCO(2) slope was not statistically significant [-2.3 (-12.2 - 1.8) vs 1.0 (-0.4 - 2.6), P > 0.05]. CONCLUSION: After 3 months of aerobic exercise, exercise capacity may improve in the CHF patients.

[Effects of early submaximal cardiopulmonary exercise test and cardiac rehabilitation for patients with acute myocardial infarction after percutaneous coronary intervention: a comparative study].

OBJECTIVE: To investigate the safety and effects of early submaximal cardiopulmonary exercise test (CPET) and cardiac rehabilitation for patients with acute myocardial infarction (AMI) after percutaneous coronary intervention (PCI). METHODS: 94 patients with AMI after PCI were randomly divided into 2 groups: exercise group undergoing anaerobic rehabilitation training based on anaerobic threshold (AT) exercise prescription for 3 months, and control group, conducting exercise according to the needs of the patients themselves. Three months later, the exercise cardiopulmonary function was evaluated. RESULTS: In the first CPET 89 patients attained their anaerobic threshold (AT) and their heart rates were lower than their target heart rates following the exercise test. The oxygen consumption at the anaerobic threshold (VO2AT) 3 months later of the exercise group was [(12.6 +/- 2.9) ml x min(-1) x kg(1)], significantly greater and that before the exercise [(10.5 x 2.9) ml x min x kg(-1), P = 0.000]. The peak oxygen uptake (VO2 pea) 3 months of the exercise group was (20 +/- 4) ml x min(-1) x kg(-1), signficantly greater then that before exercise [(14 +/- 4) ml x min(-1) x kg(-1), P = 0.000]. The LAT 3 months of the exercise group was (42 +/- 16) J x s(-1), significantly higher than that before exercise p [(33 +/- 20) J x s(-1), P = 0.000]. The workload at peak level (Lpeak) 3 months of the exercise group was (89 +/- 14) J x s(-1) significantly greater than thatbefore exercise [(66 +/- 21) J x s(-1), P = 0.000]. And the VO2pea and Lpeak of 3 months later of the control group were [(19 +/- 4) ml x min(-l) x kg(-1)) and (80 +/- 14) J x s(-1)] respectively, both significantly higher than those before exercise [(14 +/- 4) ml x min(-1) x kg(-1) and (64 +/- 21) J x s(-1), both P = 0.000]. CONCLUSION: The early submaximal CPET and cardiac rehabilitation for patients with AMI after PCI are not only safe but also can improve their exercise capacity.