With the rapid development of single‐cell sequencing technology, the Langendorff perfusion system has emerged as a common approach to decompose cardiac tissue and obtain living cardiomyocytes to study cardiovascular disease with the mechanism of cardiomyocyte biology. However, the traditional Langendorff perfusion system is difficult to master, and further, the viability and purity of cardiomyocytes are frequently unable to meet sequencing requirements due to complicated devices and manipulate processes. Here, we provide an optimized Langendorff perfusion system with a simplified and standardized operating protocol which utilizes gravity as the perfusion pressure, includes a novel method for bubbles removing and standardizes the criteria for termination of digestion. We obtained stable cardiomyocyte with high viability and purity after multiple natural gravity sedimentation. The combination of the optimized Langendorff perfusion system and the multiple natural gravity sedimentation provides a stable system for isolating adult mouse heart, which will provide higher‐quality cardiomyocytes for further experiments.

Background Monogenic hypertension describe a series of hypertensive syndromes that are inherited by Mendelian laws. Sometimes genetic testing is required to provide evidence for their diagnoses, precise classification and targeted treatment. This study is the first to investigate the clinical utility of a causative gene screening and the combined yield of gene product expression analyses in cases with suspected monogenic hypertension. Methods We performed a large-scale multi-centre clinical genetic research of 1179 expertly selected hypertensive individuals from the Chinese Han population. Targeted sequencing were performed to evaluate 37 causative genes of potential cases of monogenic hypertension. Pathogenic and likely pathogenic variants were classified using the American College of Medical Genetics guidelines. Additionally, 49 variants of unknown significance (VUS) that had relatively high pathogenicity were selected and analysed using immunoblot protein expression assays. Results 21 pathogenic or likely pathogenic variants were identified in 33 of 1179 cases (2.80%). Gene product expression analyses showed 27 VUSs harboured by 49 individuals (4.16%) could lead to abnormally expressed protein levels. Consequently, combining genetic screening with gene product expression analyses increased the diagnostic yield from 2.80% to 6.79%. The main aetiologies established were primary aldosteronism (PA; 27, 2.29%) and pheochromocytoma and paraganglioma (PPGL; 10, 0.85%). Conclusion Molecular diagnoses obtained using causative gene screening combined with gene product expression analyses initially achieved a modest diagnostic yield. Our data highlight the predominant roles of PA and PPGL. Furthermore, we provide evidence indicating the enhanced diagnostic ability of combined genetic and functional evaluation.

Blood pressure (BP) has been well documented to be associated with hearing loss previously. However, the role of blood pressure variability (BPV, representing BP fluctuation over a time period) on hearing remains unknown. We aimed to evaluate the relationship between BPV and hearing in Chinese population. We included 8646 male subjects from a population-based study (the Kailuan study). BP was measured every two years at routine physical examinations from 2006 to 2015. Based on five annual BP measurements, BPV was estimated by standard deviation of BP (SD), coefficient of the variation of BP (CV), and variation independent of mean of BP (VIM). Hearing was estimated by pure-tone average threshold (PTA) at low, intermediate, and high frequencies in the year of 2014. Regression models were used to evaluate the relationship between BPV and hearing. The results showed that PTAs and percentages of hearing loss at low, intermediate, and high frequencies grew gradually with increasing systolic SD (SSD) (p<0.05). After adjusting for multiple covariates, multivariate regression analyses demonstrated that variations of SBP (SSD, SCV, and VIMSBP) were all positively correlated with PTA at intermediate and high frequencies (p<0.05). Each SD increase in SSD, SCV, and VIMSBP was also positively associated with hearing loss at intermediate and high frequencies. No significant correlation was observed between variations of DBP and hearing. These findings suggest that increase in long-term BPV is associated with hearing and hearing loss. Trial registration number: Kailuan study (ChiCTRTNC-11001489).

Background: Trimethylamine N-oxide (TMAO) is a metabolite derived from the gut microbiota and has been reported to be correlated with cardiovascular diseases. Although TMAO is associated with the severity of coronary artery disease in subjects with coronary heart disease (CHD) history. However, the correlation between TMAO and the atherosclerotic burden in newly diagnosed cases of CHD is unknown. Methods: In this hospital-based study, we enrolled 429 individuals newly diagnosed with CHD undergoing coronary angiography. Plasma TMAO was assessed before coronary angiography. SYNTAX score was computed during coronary angiography to estimate the coronary artery atherosclerotic burden. Both linear and logistic regression analyses were conducted to explore the correlation between plasma TMAO levels and SYNTAX score in newly diagnosed CHD population. Results: The TMAO in patients with SYNTAX ≥ 33 and subjects with SYNTAX < 23 were 6.10 (interquartile range [IQR]: 3.53 to 9.15) µmol/L and 4.90 [IQR: 3.25 to 7.68] µmol/L, respectively. Linear regression adjusting for traditional risk factors showed TMAO level was positively correlated with SYNTAX score (β = 0.179; p = 0.006) in CHD population. When TMAO was added to models with traditional risk factors, the predictive value improved significantly, with the receiver operating characteristic curve (AUC) increased from 0.7312 to 0.7502 (p = 0.003). Stratified analysis showed that the correlations did not hold true for subjects who were non-smoker or with histories of diabetes. None of the stratifying factors significantly altered the correlation (all p for interaction < 0.05). Conclusions: We found a positive linear correlation between plasma TMAO and SYNTAX score among newly diagnosed CHD individuals in Chinese population.

Aldosterone‐producing adenoma (APA) is a common cause of secondary hypertension. This study aimed to explore the lncRNA‐miRNA‐mRNA competitive endogenous RNA (ceRNA) network to uncover molecular mechanism underlying APA. The mRNA and lncRNA expression data of APA and adjacent adrenal gland (AAG) from GSE60044, GSE64957 and GSE101894 were obtained from the Gene Expression Omnibus (GEO) database to analyse differentially expressed genes (DEGs) and lncRNAs (DElncs). Hub genes were identified by robust rank aggregation (RRA) and protein–protein interaction (PPI) network analysis. The miRcode and miRWalk network tools were used to construct the ceRNA network. 1526 upregulated and 1512 downregulated DEGs were identified, which are mainly enriched in extracellular matrix and Ca²⁺‐related GO terms. In the KEGG pathway analysis, Ca²⁺ signalling and the aldosterone synthesis and secretion pathways were enriched. ceRNA network included 2 lncRNAs, 9 miRNAs, and 13 mRNAs. The lncRNAs are MEG3 and LINC00115. The mRNAs included CCND1, TP53, GPRC5B, BMI1, COMMD3‐BMI1, ADAMTS15, STAT3, MMP2, SCN2B, CXCL12, HGF, FOS, and THBS1. Overall, this study conducted a ceRNA regulatory network analysis and identified that 2 lncRNAs and 13 mRNAs may contribute to the development of APA. These findings may provide novel diagnostic and intervention targets for APA.

OBJECTIVE Systolic blood pressure variability (SBPV), cumulative systolic BP (cumSBP), and systolic blood pressure trajectory (trajSBP) are major indices describing characteristics of BP changes. The aim of this study was to compare their discrimination abilities for cardiovascular (CV) events. METHODS In 51698 subjects, associations were assessed using Cox regression in the overall cohort and Framingham risk score (FRS) stratified groups. Individuals with 20% 10 -year CV risk were categorized into the low-, intermediate-, and high -risk group, respectively. Discrimination capabilities were evaluated using the area under curve (AUC), Harrell 's C index, net reclassification index (NRI), and integrated discrimination index (IDI). RESULTS Within a mean follow-up of 6.83 +/- 0.89 years, 2330 participants had CV events, and all three markers were significantly associated with CV events. TrajSBP provided the best additional discriminative value for CV events, with improvements of 1.54% in AUC%, 0.01 in Harrell 's C, 37.52% in NRI%, and 0.59% in IDI%. CumSBP had good additional discriminative capability in the intermediate to high FRS groups, but the effect sizes were smaller than those of trajSBP. Although, SBPV improved the predictive capabilities in the low-to intermediate-risk groups, the effect sizes were much smaller than those of the other indices. Sensitivity analyses excluding patients who underwent antihypertensive therapy revealed similar patterns but higher effect sizes than in the overall population. CONCLUSION TrajSBP provides the best additional discrimination capabilities based on traditional risk pro files and may assist the risk stratification and individual prediction for future CV events. (c) 2023 Hellenic Society of Cardiology. Publishing services by Elsevier B.V. This is an open access article under the CC BYNC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).