Abstract:Cardiomyopathies, a heterogeneous group of myocardial disorders, represent a principal cause of heart failure and sudden cardiac death, necessitating precise animal models to dissect their complex pathophysiology. This review systematically evaluates the key animal models used in research on dilated cardiomyopathy (DCM), hypertrophic cardiomyopathy (HCM), arrhythmogenic cardiomyopathy (ACM), and restrictive cardiomyopathy (RCM), and points out that their scientific value depends on specific research objectives. Small animal models, exemplified by mice and zebrafish, serve as the cornerstone for elucidating initial molecular pathogenesis, owing to their short breeding cycles, low husbandry costs, high-throughput screening potertial and mature genetic eding systems. In contrast, large animal models, such as swine and canines, constitute a critical platform for preclinical evaluation of pharmacological and device-based therapies, given their high fidelity to human cardiac physiology, hemodynamics, and disease progression. The analytical framework of this review is tightly coupled to disease etiology:for HCM, ACM and RCM, which are predominantly driven by monogenic mutations, this review focuses on genetically engineered models that precisely recapitulate human pathogenic mutations; for the etiologically diverse DCM, this review systematically summarizes a full spectrum of models, including those induced by genetic defects, pharmaceutical/chemical factors, and hemodynamic stress. In addition, this review highlights the unique value of spontaneous large animal models, specifically the feline HCM model and the Boxer dog ACM model, in simulating the natural history of the disease, while also discussing the limitations and specific application scenarios of each model category.

Abstract:Aim To investigate the role and molecular mechanism of proprotein convertase subtilisin/kexin type 9 (PCSK9) in the occurrence and development of atherosclerosis (As). Methods Mouse primary vascular smooth muscle cells (VSMCs) were treated with platelet-derived growth factor-BB (PDGF-BB) for different durations. Cell migration ability was evaluated by scratch assay, and the expression of phenotypic transition markers and PCSK9 was detected by Western blot. Mouse primary VSMCs were infected with Ad-PCSK9 adenovirus to examine the expression of contractile and synthetic marker proteins, as well as cell proliferation and migration capabilities. PCSK9 expression was overexpressed via adenovirus or inhibited by small interfering RNA (siRNA), and the expression levels of nuclear factor-κB (NF-κB), Krüppel-like factor 4 (KLF4), and myocardin (MYOCD) were detected. Immunohistochemistry, immunofluorescence, Oil Red O staining, and hematoxylin-eosin (HE) staining were used to analyze the expression of related molecules, lipid deposition, and plaque area in aortic plaques of VSMC-specific PCSK9 overexpression (PCSK9^sm OE) mice and PCSK9 conditional knockout (PCSK9^flox/flox) mice. Results PCSK9 was colocalized with VSMCs in the medial layer and atherosclerotic plaques. Compared with contractile VSMCs in the medial layer, PCSK9 expression was significantly elevated in synthetic VSMCs within the plaques. PDGF-BB upregulated PCSK9 in a time-dependent manner, along with the synthetic phenotypic marker proteins osteopontin (OPN) and epiregulin (EREG), while downregulating the contractile marker proteins smooth muscle myosin heavy chain (SMMHC), α-smooth muscle actin (α-SMA), calponin, and smooth muscle 22α (SM22α), and enhanced cell migration capacity. Ad-PCSK9 promoted the phenotypic transformation of VSMCs from contractile to synthetic phenotype, significantly enhanced DNA replication activity and cell migration ability, downregulated MYOCD expression, and upregulated KLF4 and NF-κB p65 expression. Conversely, siRNA-mediated PCSK9 inhibition showed opposite effects. Histological analysis revealed that VSMC-specific overexpression of PCSK9 promoted aortic plaque formation and increased plaque instability in atherosclerotic mice. Conclusions PCSK9 can induce VSMC phenotypic transformation from the contractile to synthetic phenotype, exacerbating the atherosclerotic lesions in mice. This mechanism may involve the signaling crosstalk among NF-κB, KLF4, and MYOCD.

Abstract:Aim To investigate the role of microRNA-101-3p (miR-101-3p) in a mouse model of heart failure after left anterior descending branch (LAD) coronary artery ligation-induced myocardial infarction (MI), and to analyze its association with Ras-related C3 botulinum toxin substrate 1/p21 activated kinase 1 (RAC1/PAK1) signaling, endothelial-to-mesenchymal transition (EndMT), and myocardial fibrosis. Methods Heart failure after MI was induced by LAD ligation in mice, which were then randomly assigned to the sham, MI model, miR-101-3p agomir (miR-101-3p-Ago) treatment, RAC1 inhibitor (NSC23766) treatment, combined treatment, and metoprolol positive-control groups. Four weeks after surgery, transthoracic echocardiography was performed to assess left ventricular ejection fraction (LVEF), fractional shortening (FS), left ventricular end-diastolic/systolic diameter (LVEDD/LVESD), and wall thickness. Hematoxylin-eosin, Masson and wheat germ agglutinin (WGA) staining were used to evaluate myocardial structural remodeling and fibrosis. Immunofluorescence co-staining for VE-cadherin/α-smooth muscle actin (α-SMA) and CD31/vimentin was performed to assess EndMT-related phenotypes. Western blot was used to detect the expression of RAC1, PAK1 and EndMT-related proteins. Transmission electron microscopy (TEM) was employed to observe myocardial ultrastructural changes in cardiomyocytes. Results Compared with the Sham group, cardiac function (LVEF, FS) in the model group was significantly decreased, and ventricular remodeling and myocardial fibrosis were aggravated (all P＜0.05). Meanwhile, the RAC1/PAK1 pathway was activated in myocardial tissue, accompanied by significantly enhanced EndMT-related phenotypes (all P＜0.01). Treatment with either miR-101-3p-Ago or the RAC1 inhibitor alone significantly improved cardiac function, suppressed RAC1/PAK1 activation, and attenuated EndMT-related abnormalities (P＜0.05). The combined intervention exerted the strongest protective effect, reducing the myocardial fibrotic area by more than 60% and significantly restoring functional indices such as LVEF (P＜0.01). Moreover, its anti-fibrotic and cardioprotective effects on key parameters were significantly superior to those of either monotherapy (P＜0.05), indicating a clear synergistic benefit. TEM findings further suggested that the combined intervention partially ameliorated myocardial ultrastructural injury. Conclusion miR-101-3p agomir improves myocardial fibrosis and cardiac function in mice after myocardial infarction by inhibiting RAC1/PAK1mediated EndMT, suggesting that it may serve as a potential molecular target for anti-fibrotic therapy after myocardial infarction.

Abstract:Aim To explore the predictive value of hematocrit (HCT), plasma D-dimer combined with fibrinogen (FIB) for the risk of lower extremity deep venous thrombosis (DVT) in patients with hemorrhagic stroke. Methods Clinical data were retrospectively collected from 210 patients with hemorrhagic stroke admitted to the Second People's Hospital of Hefei from January 2023 to June 2025, including 81 patients with DVT (DVT group) and 129 patients without DVT (non-DVT group). 120 patients with hemorrhagic stroke admitted during the same period were enrolled as the external validation cohort. Multivariate Logistic regression was employed to identify influencing factors for DVT in hemorrhagic stroke, and ROC curves together with calibration curves were applied to establish and validate the predictive model. Results Patients in the DVT group were older, with higher admission NIHSS scores, longer bedridden duration, and elevated levels of HCT, FIB and D-dimer compared with the non-DVT group (P＜0.05). There was no statistically significant difference in other baseline data (P＞0.05). Multivariate Logistic regression analysis showed that elevated NIHSS scores (OR＝1.8,5%CI:1.066～1.324), prolonged bedridden duration (OR＝1.9,5%CI:1.330～1.851), increased HCT (OR＝1.9,5%CI:1.088～1.343), FIB (OR＝2.3,5%CI:1.523～4.905) and D-dimer (OR＝2.0,5%CI:1.219～3.617) at admission were independent risk factors for DVT in patients with hemorrhagic stroke (all P＜0.05); the combined detection of admission NIHSS score, bedridden duration, HCT, FIB and D-dimer yielded a higher AUC for predicting DVT than any single indicator, with statistically significant differences (all P＜0.05). Conclusion Elevated admission NIHSS score, prolonged bedridden duration, and increased HCT, FIB and D-dimer levels are independent risk factors for DVT in patients with hemorrhagic stroke; the combination of these indicators presents high predictive efficacy for DVT risk.

Abstract:Aim To investigate the association between the longitudinal changes of C-reactive protein-triglyceride-glucose index (CTI) and the longitudinal trajectories of relative fat mass (RFM) with the risk of new-onset stroke in middle-aged and elderly populations. Methods The study used data from the China Health and Retirement Longitudinal Study (CHARLS) for analysis. Based on two measurements of CTI in 2011 and 2015, the K-mean clustering was used to identify the categories of CTI changes. Based on three measurements of RFM in 1,3 and 2015, latent class mixed-growth modeling was applied to fit the RFM trajectory. Multivariate Logistic regression was used to assess the risk effects of CTI categories, RFM trajectories, and their combination on new-onset stroke. Relative excess risk analysis was conducted to evaluate the interaction between CTI categories and RFM trajectories. ROC curve analysis was performed to evaluate the discriminatory power of CTI categories and RFM trajectories for new-onset stroke. Results Of the 4 823 participants meeting the inclusion and exclusion criteria, 393 developed new-onset stroke during a median follow-up of 3 years. After adjusting for confounding factors, Logistic regression analysis showed that compared to category 1 with consistently low CTI levels, category 3 with consistently high CTI levels had a 70% increased risk of new-onset stroke (OR＝1.0,5%CI:1.24～2.34), while the RFM high stability group (trajectory 3) had a 62% increased risk compared to the low stability group (trajectory 1) (OR＝1.2,5%CI:1.04～2.64). Joint analysis showed that participants in category 3 and trajectory 3 had a significantly higher risk of new-onset stroke than those in category 1 and trajectory 1 (OR＝3.3,5%CI:1.98～6.96). The relative excess risk interaction analysis revealed that there was a significant additive interaction and synergistic effect between CTI categories and RFM trajectories. ROC curve analysis showed that CTI categories combined with RFM trajectories had a certain discriminatory power for new-onset stroke (AUC＝0.617). Conclusion The sustained high levels of CTI and RFM are significantly associated with an increased risk of new-onset stroke, and continuous monitoring of CTI and RFM is of great significance for the prevention and treatment of new-onset stroke.

Abstract:Aim To explore the changes of non-invasive hemodynamic parameters in young and middle-aged patients with hypertension and their relationship with carotid arterial lesions. Methods A total of 689 hypertensive patients admitted to our hospital from October 2021 to April 2024 were enrolled as the research subjects. Patients were divided into hypertension grade 1 (systolic blood pressure (SBP):140～159 mmHg; diastolic blood pressure (DBP):90～99 mmHg), hypertension grade 2 (SBP:160～179 mmHg; DBP:100～109 mmHg) and hypertension grade 3 (SBP:≥180 mmHg; DBP:≥110 mmHg) according to their levels of SBP and DBP. The severity of carotid artery lesions was evaluated using the Crouse score, with the grading criteria as follows:non-lesion (0 point:intima-media thickness (IMT) ＜1 mm), IMT thickening (1 point:IMT 1～1.2 mm), plaque formation (2 points:IMT 1.2～1.5 mm), mild stenosis (3 points:IMT 1.5～2.0 mm) and severe stenosis (4 points:IMT＞2.0 mm). In addition, patients were divided into the carotid artery lesion group (IMT≥1 mm, Crouse score≥1 point) and the non-lesion group (IMT ＜1 mm, Crouse score ＜1 point) according to IMT values. Multivariate Logistic regression model was used to identify risk factors for carotid artery lesions in hypertensive patients. Multiplicative model and additive model were used to analyze the interaction between age and non-invasive hemodynamic parameters on carotid arterial lesions. Additionally, restricted cubic spline model was used to examine the dose-response relationship between non-invasive hemodynamic parameters and carotid arterial lesions in hypertensive patients. Results In young and middle-aged hypertensive patients, the proportions of severe coronary artery stenosis (Gensini score ≥ 50 points) and Crouse score≥ 2 points, as well as thoracic fluid content (TFC), stroke volume variation (SVV), systemic vascular resistance (SVR), systemic vascular resistance index (SVRI), acceleration index (ACI), mean arterial pressure (MAP), 24-hour mean systolic blood pressure (24h-mSBP), 24-hour mean diastolic blood pressure (24h-mDBP), 24-hour systolic blood pressure variability (24hSBPV), and 24-hour diastolic blood pressure variability (24hDBPV) all showed a gradual increase from hypertension grade 1 to grade 3 (P＜0.05). Compared with the non-lesion group, age and the above-mentioned indicators were significantly increased in the carotid artery lesion group (P＜0.05). Middle-age, grade 3 hypertension, TFC＞32 kΩ－1, SVR＞1 534 dyne·s/cm⁵, ACI＞1,4h-mSBP＞167 mmHg and 24hSBPV＞12.9 mmHg were independent risk factors for carotid arterial lesions. Age exhibited multiplicative interaction (OR＞1) and additive interaction (RERI＞0, AP＞0, SI＞1) with TFC, SVR and ACI on carotid arterial lesions. Levels of TFC, SVR, and ACI exhibited a non-linear dose-response relationship with carotid arterial lesions in young hypertensive patients (P_non-linearity＝0.2,0.6,0.028) and middle-aged hypertensive patients (P_non-linearity＝0.3,0.7,0.015). Conclusion In young and middle-aged hypertensive patients, the levels of TFC, SVR and ACI all increase with the elevated blood pressure grade, as well as the elevation of each indicator is an independent risk factor for carotid arterial lesions.

Abstract:Aim To investigate the predictive value of a backpropagation artificial neural network (BPNN) model based on novel inflammatory factors for the severity grading of coronary artery lesions in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods A total of 234 patients with acute STEMI admitted to the Cardiac Center of First Affiliated Hospital of Xinjiang Medical University from January 2022 to October 2024 were enrolled as study subjects. They were randomly divided into a training set (164 cases) and a test set (70 cases) at a 7∶3 ratio.Based on the severity grading of coronary artery lesions (SYNTAX score), the training set was further classified into a low-risk group (0~22 points) with 54 cases (32.9%), a medium-risk group (23~32 points) with 62 cases (37.8%), and a high-risk group (≥33 points) with 48 cases (29.3%). General clinical data, serum inflammatory factors (high sensitivity C-reactive protein (hs-CRP), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), and novel inflammatory factors (neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), pan-immune inflammation value (PIV), systemic immune-inflammation index (SII) were compared among three groups of acute STEMI patients in the training set. Multivariate ordinal Logistic regression analysis was used to identify independent predictors of coronary artery lesion severity grading in acute STEMI patients. Random forest models and BPNN models were constructed, and the predictive performance of the models was evaluated using the area under the curve (AUC) from ROC curve analysis, as well as accuracy, sensitivity, and specificity calculated from the confusion matrix. Results Multivariate ordinal Logistic regression analysis showed that the triglyceride-glucose index (TyG), hs-CRP, and PIV were independent predictors of coronary artery lesion severity grading in acute STEMI patients (P＜0.05). Based on these three independent predictors, random forest models and BPNN models were constructed to predict the severity grading of coronary artery lesions in acute STEMI patients. The random forest model showed that when 2 654 decision trees were generated, the error rates for predicting high-risk, medium-risk, low-risk, and overall out-of-bag data stabilized at 15.3%, 19.2%, 30.5% and 21.4%, respectively, indicating stable accuracy of the model. Hyperparameter tuning of the BPNN model was performed using 5-fold cross-validation and grid search, and the optimal parameter combination was identified as follows:the network topology of the BPNN was 3-4-2-3, the maximum number of iterations was 10⁷, and the learning rate was 0.5, at which the average accuracy reached a maximum of 84.3%. Finally, after 201 935 iterations of weight updates, the loss function reached a minimum value of 20.659 917. In the training set, the AUC of the BPNN model for predicting high-risk, medium-risk, and low-risk was increased by 6.5%, 8.9% and 8.3%, respectively, compared with the random forest model (all P＜0.05). In the test set, the AUC of the BPNN model for predicting high-risk, medium-risk, and low-risk was increased by 5.2%, 9.4% and 13.4%, respectively, compared with the random forest model (all P＜0.05). In both the training and test sets, the sensitivity, specificity, and accuracy of the BPNN model for predicting high-risk, medium-risk, and low-risk were higher than those of the random forest model. Conclusion Both the BPNN model and the random forest model constructed based on TyG, hs-CRP, and PIV exhibit good predictive efficacy for the severity grading of coronary artery lesions in acute STEMI patients, and the BPNN model outperforms the random forest model.

Abstract:Aim To establish a single-step density gradient centrifugation method for isolating cholesterol-rich non-high density lipoprotein (CR-nHDL) particles and investigate the effect of oxidized CR-nHDL on foam cell formation. Methods A single-step density gradient centrifugation method using ioversol as the density medium was established to isolate CR-nHDL with a density of less than 1.008 g/mL from postprandial plasma of healthy volunteers. The density of the fraction was measured, and the cholesterol content was detected enzymatically. The lipoprotein content in the particles was analyzed by Western blot and proteomics. The isolated CR-nHDL was oxidized and co-cultured with mouse bone marrow-derived macrophages (BMDM), and its effect was compared with that of oxidized low density lipoprotein (ox-LDL). The ability of oxidized CR-nHDL to induce macrophage foam cell formation was assessed by measuring intracellular lipid accumulation. Results This method successfully isolated CR-nHDL with a density of less than 1.008 g/mL. Western blot results showed that this fraction contained apolipoprotein B100 (ApoB100) and apolipoprotein B48 (ApoB48) but not apolipoprotein AⅠ (ApoAⅠ). The results of proteomic analysis showed that there was no significant difference in the major apolipoprotein components between CR-nHDL prepared by ioversol method and low density lipoprotein (LDL) prepared by classical two-step method; lipidomics analysis results showed that the cholesterol ester content in the CR-nHDL prepared by the ioversol method was significantly higher than that in the LDL prepared by the classical method, which suggested that this method was more efficient in enriching cholesterol-containing lipoprotein particles, and the oxidized CR-nHDL obtained from this method could promote lipid accumulation in macrophage derived foam cells more than ox-LDL. Conclusion The single-step density gradient centrifugation of ioversol can quickly isolate CR-nHDL, and its oxidized form has a stronger ability to promote macrophage foam cell formation.

Abstract:Atherosclerotic cardiovascular disease is one of the major fatal diseases in the world, and its pathological basis is chronic inflammation in essence. A large amount of evidence shows that inflammation plays a key role in the occurrence and development of atherosclerosis, in which immune cells and inflammatory signal pathways jointly regulate the inflammatory microenvironment in the blood vessels, thereby accelerating the process of disease. However, current targeted anti-inflammatory applications are still limited by issues such as limited targets, lack of inflammatory monitoring indicators, and treatment safety. This article focuses on the inflammatory mechanism of atherosclerotic cardiovascular disease, systematically reviews the regulatory role of immune cells and inflammatory signaling pathways on its inflammatory process, and discusses the potential clinical transformation strategies from multiple dimensions such as the clinical application of new inflammatory markers and targets, precision intervention technology, and lifestyle management, which provides a new idea for building an accurate prevention and treatment system targeting inflammation.

Abstract:GATA4, a key member of the GATA zinc finger family, plays a crucial role in cellular senescence. GATA4 affects chronic inflammation, oxidative stress, cell cycle progression, apoptosis, and metabolic alterations to facilitate the development of cellular senescence. GATA4 promotes vascular endothelial cell senescence and drives the development of coronary atherosclerotic heart disease (CAD); GATA4 facilitates pathological myocardial tissue remodeling, hypertrophy, and the progression of heart failure; GATA4 accelerates lens epithelial cell senescence, hastening the onset and progression of age-related cataract (ARC). Conclusively, GATA4 plays an important role in cellular senescence and related cardiovascular diseases, and can serve as a potential target for the diagnosis and treatment of age-related diseases.

Abstract:Ischemic heart disease (IHD) is the leading cause of death worldwide. Although coronary intervention has significantly improved the prognosis of patients with obstructive coronary heart disease, nearly three-quarters of patients with angina pectoris are diagnosed with ischemia with non-obstructive coronary artery disease (INOCA). Among these, coronary microcirculation dysfunction (CMD) is the primary mechanism underlying angina pectoris and myocardial ischemia in INOCA patients, accounting for approximately two-thirds of all INOCA cases. Notably, the prevalence rate in women (ranging from 56% to 82%) is significantly higher than that in men. CMD is not only closely associated with adverse cardiovascular events such as heart failure, myocardial infarction, and sudden cardiac death, but also contributes to the development of coronary atherosclerosis. Additionally, it leads to a decline in patients’ quality of life, with nearly half of CMD patients experiencing anxiety and depression. However, only less than 7% of CMD patients in China receive appropriate treatment, highlighting prominent challenges in diagnosis and management. Based on the latest research progress, this article systematically reviews the multi-mechanistic pathogenic basis, non-specific clinical manifestations, diagnostic difficulties, and current treatment status of CMD. It aims to provide references for clinical precise diagnosis and treatment, and clarify the future research directions in this field.

Abstract:Atherosclerosis (As) serves as the pathological basis of various cardiovascular and cerebrovascular diseases. The pathogenesis of As involves intricate and interconnected pathological mechanisms, in which multiple molecular pathways interact synergistically via complex crosstalk to collectively drive disease initiation and progression. Traditional Chinese medicine (TCM) compounds exhibit unique advantages in the prevention and treatment of As by modulating multiple pathological processes, including vascular endothelial function, lipid metabolism, immune and inflammatory responses, oxidative stress, gut microbiota, extracellular vesicles, and macrophage autophagy. Based on the pathological mechanisms of As, this article systematically elucidates the characteristics of TCM compounds in prevention and treatment of As, aiming to provide more comprehensive theoretical evidence for As research and intervention.