The porcine reproductive and respiratory syndrome virus (PRRSV) is a highly contagious pathogen. Viral infections often enhance their replication by modulating the structure and expression of host genes. However, it remains unclear whether PRRSV employs a similar mechanism to achieve self-replication. To address this question, the current study combined assay for transposase accessible chromatin sequencing (ATAC-seq) and ribonucleic acid (RNA) sequencing (RNA-seq) to identify accessible chromatin regions and key host genes associated with PRRSV infection. By comparing the PRRSV-infected group with the control group, we initially detected 8664 differentially accessible chromatin regions and 4037 differentially expressed genes. Motif analysis of these differential chromatin regions revealed several potential cis-regulatory elements containing binding sites for transcription factors. Further integration of ATAC-seq and RNA-seq results identified 1352 overlapping genes between the PRRSV-infected and control groups. A significant positive correlation between differential gene expression and chromatin accessibility signals suggests that chromatin remodeling may drive transcriptional changes during infection. Protein–protein interaction (PPI) network analysis highlighted candidate genes potentially associated with PRRSV infection in hosts, such as IL1B, CCL20, CXCL10, CSF3, etc. Given their potential association with the infection mechanism, these genes could serve as candidate targets for the future development of prophylactic vaccines and therapeutic strategies. Additionally, several signaling pathways that may regulate immune and inflammatory responses were significantly enriched in our ATAC-seq and RNA-seq analyses. These findings provide valuable insights into the molecular mechanisms underlying PRRSV infection and pave the way for developing more effective preventive and treatment measures.

Porcine reproductive and respiratory syndrome virus is a major pathogen that causes massive economic losses in the global swine industry. Lysine propionylation and malonylation are metabolism-sensitive post-translational modifications, yet their coordinated regulatory roles during PRRSV pulmonary infection remain unknown. This study combined quantitative proteomics, propionylome and malonylome to characterize host molecular alterations between healthy and PRRSV-infected porcine lung tissues (three piglets per group). Quantitative proteomics identified 1467 significantly downregulated proteins and only 129 upregulated proteins, indicating a profound host protein shutoff during PRRSV pulmonary infection. The two acyl modifications displayed opposite regulatory patterns independent of global protein expression changes: 51 propionylation sites (24 proteins) were upregulated with only 1 downregulated, while 37 malonylation sites (30 proteins) were downregulated and only 5 upregulated. Functional enrichment and PPI network analysis revealed clear functional divergence: hyper-propionylated hub proteins were exclusively enriched in mitochondrial energy metabolism pathways, whereas hypo-malonylated core proteins were mainly involved in lipid metabolism and cell fate regulation. Integrated multi-omics analysis confirmed that protein expression and lysine acylation constitute two coordinated but independent regulatory layers, with histone H4 identified as a candidate target of competitive propionylation/malonylation. This work reports the comprehensive landscape of lysine propionylation and malonylation upon PRRSV infection, reveals a dual-PTM remodeling strategy for viral hijacking of host homeostasis, and provides candidate targets for future functional investigation and antiviral development.

Growth traits, as core economic indicators in pig breeding, are closely associated with production costs, rearing duration, and final carcass quality and have thus consistently been a major focus of genetic improvement. This study aimed to identify candidate genes affecting Age to 120 kg live weight (AGE120), Backfat thickness at 120 kg (BF120), and Loin muscle depth at 120 kg (LMD120) in pigs. Ear tissue samples were collected from 3364 healthy adult pigs (including 558 boars and 2805 sows) from three breeds: Large White, Landrace, and Duroc. Genotyping was performed using an 80 K functional site array, and quality-controlled SNP (Single-Nucleotide Polymorphism) loci were subjected to genotype imputation, resulting in 15,447,611 loci obtained. Genome-wide association studies (GWASs) for Age to 120 kg live weight, Back fat thickness at 120 kg, and Loin muscle depth at 120 kg were conducted using a mixed linear model in Genome-wide Complex Trait Analysis (GCTA). Genes located within 500 kb upstream and downstream of significant GWAS loci were extracted using the biomaRt package in R. Furthermore, colocalization analysis was performed using expression Quantitative Trait Locus (eQTL) data of 34 tissues from the PigGTEx database to identify genes that share the same causal variant as the GWAS signals. Through integrated GWAS and eQTL colocalization analysis, in addition to five previously reported genes associated with pig growth traits (TAF11, ZC3HAV1L, ANKS1A, USP20, and TBC1D1), a set of novel, high-confidence candidate genes was identified: ZNF215, UBE2Z, HOXB7, SARDH, ADAMTSL2, ATP6V0A4, RPL10A, PGM2, and RELL1. These findings enrich our understanding of the genetic architecture underlying growth traits in pigs at heavy body weights and provide an important foundation for subsequent functional validation and molecular breeding applications.

CONTEXT:Germline CACNA1H mutations have been associated with primary aldosteronism (PA). Here, we described 3 newly discovered CACNA1H variants and explored the possible pathogenic mechanisms as well as therapeutic strategies. DESIGN:Targeted next-generation sequencing of 10 PA causative genes was performed in 305 PA patients. Variants with possible clinical significance in CACNA1H were identified. Functional analysis was performed in vitro including electrophysiology, steroid hormone production, and transcriptome. Responses to mibefradil and nifedipine were evaluated. RESULTS:We described 3 heterozygous germline CACNA1H variants (E282K, V213M, and S1249R) in 4 independent cases of PA. Recurrent E282K mutation was identified in 1 patient with severe early-onset PA and 1 patient with bilateral adrenal hyperplasia (BAH). Electrophysiological analysis of mutant Cav3.2 channels revealed significant gain-of-function changes in the voltage-dependent activation and Ca2+ current properties for Cav3.2E282K, while ambiguous for Cav3.2S1249R and Cav3.2V213M. Moreover, stable expression of the 3 CACNA1H variants in H295R cells led to increased aldosterone production and upregulated expression of genes involved in steroidogenic enzymes, both basally and after K+ treatment. T-type calcium channel blocker showed more significant inhibition of aldosterone production in mutant Cav3.2 cells compared to L-type calcium channel blocker. CONCLUSIONS:Our study suggests that Cav3.2E282K is a gain-of-function mutation and T-type calcium channel blockers may be efficient in controlling aldosterone levels for PA patients carrying mutant Cav3.2.

Adrenocortical carcinoma (ACC) is a rare but aggressive malignancy with poor survival and limited treatment options. To comprehensively characterize its molecular landscape and identify clinically relevant subtypes, we performed an integrated genomic analysis - including whole-exome sequencing, RNA sequencing, and copy number variation profiling - on 61 Chinese patients with ACC. We identified recurrent mutations in TP53 (25%), CTNNB1 (15%), ZNRF3 (10%), and MEN1 (8%). Unsupervised clustering of transcriptomic data revealed four distinct molecular subtypes: cortisol-driven (CD, 14%), immune-suppressed (IS, 40%), cell cycle-altered (CCA, 22%), and immunomodulatory (IM, 24%). The CD subtype exhibited steroidogenic pathway activation; the IS subtype showed T cell receptor downregulation and the worst disease-free survival; the CCA subtype was marked by chromosomal instability and cell cycle gene overexpression; and the IM subtype displayed enriched immune signaling and favorable outcomes. Copy number analysis further uncovered focal amplifications (e.g. TERT, CDK4) and HLA-II deletions. This study establishes a novel molecular classification of ACC, providing a framework for subtype-specific therapeutic strategies, such as CDK4/6 inhibition for CCA and immunotherapy for IM tumors, while highlighting the clinical challenges of immune-cold IS tumors.

Xiangyang Black (XYB) is a valuable indigenous pig breed from Hubei Province, China, known for its high meat quality, disease resistance, and adaptability to the local conditions. To explore the genetic diversity, population structure, and selection signatures of XYB in the context of Chinese and Western pig breeds, we performed whole-genome resequencing on 15 pig breeds, involving 225 individuals. After quality control, 20,479,203 high-quality single-nucleotide polymorphisms (SNPs) were retained for subsequent analysis. Genetic diversity analysis revealed that XYB exhibited relatively high genetic diversity, with an observed heterozygosity (Ho) of 0.38 and a nucleotide diversity (pi) of 0.35 and the inbreeding coefficient based on runs of homozygosity (FROH) ranged from 0.001 to 0.090, indicating its strong potential for genetic improvement and conservation. Population structure analyses—including neighbor-joining tree, principal component analysis, and admixture analysis—revealed a distinct genetic structure and confirmed the distinct genetic status of XYB at the molecular level. A combination of three complementary methods—fixation index (Fst), nucleotide diversity (θπ) ratio, and cross-population extended haplotype homozygosity (XP-EHH)—revealed 1,080 putative selective sweep regions (covering approximately 76.1 Mb of the genome) harboring 951 candidate genes in the Xiangyang Black pig breed compared to Western breeds. Functional annotation showed that these genes were enriched in pathways related to meat quality (e.g., FABP2, PPARG, C/EBPα, and THRSP), reproduction (e.g., GNRH1, CENPE, and CCDC112), and disease resistance (e.g., CCL17, CCL22, and CX3CL1). Our results provide insights into the genetic basis of phenotypic traits in XYB pigs and offer a theoretical foundation for their conservation, breeding, and genetic improvement.

High-throughput genome sequencing and genotyping have significantly accelerated genetic research. However, the high cost of whole-genome sequencing (WGS) remains a barrier to large-scale studies like genome-wide association studies (GWAS) and genomic prediction. Genotype imputation offers a cost-effective alternative by inferring unobserved variants from lower-density data using haplotype reference panels. In this study, we present the updated Pig Haplotype Reference Panel (PHARP) 4.0, comprising 6449 pig genomes from 154 breeds. PHARP 4.0 encompasses 50.3 million SNPs and 5.8 million indels, making it the largest and most diverse pig reference panel to date. PHARP 4.0 demonstrated superior imputation accuracy compared to existing panels (SWIM, AHC, AGIDB, and PGRP), achieving concordance rates (CR > 0.99) and correlation coefficients (R² > 0.98) in European breeds and improved accuracy in Chinese Jinhua pigs (CR = 0.936, R² = 0.924) when imputing from 80 K SNP chip data to whole-genome sequencing (WGS). We further optimized an RNA-seq-based imputation pipeline by incorporating multiple breeds and applying a 6× sequencing depth filter, achieving CR > 0.95 and R² > 0.90 in European breeds, and a CR of 0.93 with an R² = 0.92 in Chinese Jinhua pigs. Additionally, increasing the specific reference panel size to approximately 400 samples improved the imputation of rare variants. Utilizing PHARP 4.0, we successfully imputed low-density SNP chip data for two GWAS, identifying significant SNPs likely representing causal variants. Overall, PHARP 4.0 serves as a valuable resource for advancing pig genetic research and supporting breeding programs. PHARP 4.0 is an updated pig haplotype reference panel with 6449 genomes from 154 breeds. It demonstrates superior imputation accuracy from chip to WGS data, enables an optimized RNA-seq imputation pipeline, and successfully identifies novel causal variants in GWAS.

CONTEXT:The ubiquitin-specific protease 8 (USP8) gene mutations are the most common driver changes in Cushing disease (CD). However, few studies have investigated the association between USP8 mutation and recurrence, and the results have been inconclusive. OBJECTIVE:To identify the predictors of recurrence and evaluate the prognostic role of USP8 mutation. METHODS:One hundred and seven patients with pathologically confirmed corticotroph adenomas were included. Somatic USP8 mutations were identified using Sanger sequencing. Recurrence predictors were estimated using multivariate Cox models, followed by receiver operating characteristic curves and Kaplan-Meier analysis. RESULTS:Thirteen patients (12.6%) experienced recurrence, with a mean follow-up period of 65 months after surgery. The recurrence rate was significantly higher in USP8-mutated tumors than in USP8 wildtype tumors (26.5% vs 5.8%; P = .009). Multivariate Cox models revealed that USP8 mutation, high postsurgical morning serum cortisol (MSC), and high 1-mg dexamethasone suppression test (DST) were associated with an increased 5-year recurrence risk. Furthermore, Kaplan-Meier survival analysis showed that patients with USP8 mutation, combined with either high postsurgical MSC (>2.5 μg/dL) or high 1-mg DST (>0.78 μg/dL), were more prone to recurrence (log-rank P < .001). The negative predictive values were 98% and 100%, while the positive predictive values improved from 33% to 55% and from 47% to 86%, respectively. CONCLUSION:Our study corroborates USP8 mutational status in combination with postsurgical MSC or 1-mg DST as independent predictors of long-term remission, highlighting their potential role in stratifying patients at risk for suboptimal outcomes.

Jianli pig (JL) is a representative Chinese local pig breed with a unique fruity flavor and excellent meat quality. However, the reasons for the unique fruity flavor of Jianli pork are still unclear. This study explored the potential genetic mechanisms by performing an integrative analysis of the flavoromics, lipidomics, and transcriptomics of the longissimus thoracis (LT) from JL pigs and Duroc × Landrace × Yorkshire pigs (DLY). The results indicated that the relative abundance of Acetic acid butyl ester and 3-Carene flavor compounds characterized by sweet and fruity aroma in JL pork was higher compared with DLY pigs. Lipidomics results showed that 16-carbon and 18-carbon fatty acids are important lipid precursors for the flavor of JL pork. Moreover, two clusters of functional genes correlated with 3-Carene (DCHS2, NRXN1, JAKMIP3, and TRO) and Acetic acid butyl ester (WFIKKN2, CES3, and IYD) were identified. This study enriched the limited understanding of the unique fruity flavor formation in JL pigs, provided a theoretical basis for the breeding of high-quality pig breeds, and the processing of flavorful meat products.

Pork serves as a significant meat commodity, with intramuscular fat (IMF) content being a critical determinant of its quality. However, the epigenetic mechanism of porcine IMF deposition is still unclear. This study integrated proteomics and lactylation profiles from the longissimus thoracis (LT) muscles of pigs with extremely high (IMF_H) and extremely low (IMF_L) IMF content to clarify the association between lactylation and porcine fat deposition. Furthermore, an intramuscular preadipocyte induction and differentiation model was conducted to elucidate the changes in lactylation during adipocyte differentiation. Finally, the regulatory role of lactylation in adipocyte differentiation was explored by modulating lactate production during the induction and differentiation of preadipocytes. Proteomic analysis revealed significantly increased expression of key lipid metabolism related proteins (FASN, APOA4, FABP4, ACLY, PLIN1) in IMF_H pig muscle tissues compared with IMF_L tissues, along with substantial activation of lipid metabolism pathways. Lactylation profiling identified 95 differential lysine sites across 56 proteins, with most showing lower lactylation levels in the IMF_H group. The integrative omics analysis revealed differences in lactylation profiles in porcine LT tissues with varying efficiencies of IMF deposition, highlighted PGK1, PKM, and PYGM as central lactylation-modified proteins in porcine fat deposition regulation. Further in vitro study proved that lactate-mediated lactylation inhibited adipogenic differentiation of porcine intramuscular preadipocytes through PPARγ signaling pathway. This study clarified the changes in the lactylation profile in porcine LT tissues with varying efficiencies of IMF deposition, and demonstrated that lactate-mediated lactylation inhibits the PPARγ signaling pathway and the adipogenic differentiation of porcine intramuscular preadipocyte. This study provided a new insight to understanding the epigenetic regulation mechanisms of lipid deposition in pigs.

Understanding the molecular and cellular mechanisms underlying complex traits in pigs is crucial for enhancing genetic gain via artificial selection and utilizing pigs as models for human disease and biology.Here,we conducted comprehensive genome-wide association studies(GWAS) followed by a cross-breed meta-analysis for 232 complex traits and a within-breed met a-analysis for 12 traits,using 28.3 million imputed sequence variants in 70 328 animals across 14 pig breeds.We identified 6878 quantitative trait loci(QTL) for 139 complex traits.Leveraging the Pig Genotype-Tissue Expression resource,we systematically investigated the biological context and regulatory me chanisms behind these trait-QTLs,ultimately prioritizing 14 829 variant-gene-tissue-trait regulatory circuits.For instance,rs344053754 regulates UGT2B31 expression in the liver and intestines,potentially by modulating enhancer activity,ultimately influencing litter weight at weaning in pigs.Furthermore,we observed conservation of certain genetic and regulatory mechanisms underlying complex traits between humans and pigs.Overall,our cross-breed meta-GWAS in pigs provides invaluable resources and novel insights into the genetic regulatory and evolutionary mechanisms of complex traits in mammals.

Post-translational modifications (PTMs) of proteins, as the core mechanism for dynamically regulating follicular development, affect the maintenance of mammalian fertility by precisely coordinating granulosa cell–oocyte interaction, metabolic reprogramming, and epigenetic remodeling. Dysregulation of these modifications directly contributes to major reproductive diseases, including polycystic ovary syndrome (PCOS) and premature ovarian insufficiency (POI). Post-translational modifications regulate follicular development through intricate mechanisms. Thus, this review systematically synthesizes recent advances in PTMs, encompassing traditional ones such as phosphorylation, ubiquitination, and acetylation, alongside emerging modifications including lactylation, SUMOylation, and ISGylation, thereby constructing a more comprehensive PTM landscape of follicular development. Furthermore, this study dissects the molecular interaction networks of these PTMs during follicular activation, maturation, and ovulation, and uncovers the common mechanisms through which PTM dysregulation contributes to pathological conditions, including hyperandrogenism in PCOS and follicular depletion in POI. Finally, this review ultimately provides a theoretical basis for improving livestock reproductive efficiency and precise intervention in clinical ovarian diseases.

Background Jianli pig, a renowned indigenous breed in China, has the characteristics of a two-end black (TEB) coat color, excellent meat quality, strong adaptability and increased prolificacy. However, there is limited information available regarding the genetic diversity, population structure and genomic regions under selection of Jianli pig. On the other hand, the genetic mechanism of TEB coat color has remained largely unknown. Results In this study, the whole genome resequencing of 30 Jianli pigs within a context of 153 individuals representing 13 diverse breeds was performed. The population structure analysis revealed that Jianli pigs have close genetic relationships with the Tongcheng pig breed, their geographical neighbors. Three methods (observed heterozygosity, expected heterozygosity, and runs of homozygosity) implied a relatively high level of genetic diversity and, a low inbreeding coefficient in Jianli compared with other pigs. We used Fst and XP-EHH to detect the selection signatures in Jianli pigs compared with Asian wild boar. A total of 451 candidate genes influencing meat quality ( CREBBP , ADCY9 , EEPD1 and HDAC9 ), reproduction ( ESR1 and FANCA ), and coat color ( EDNRB , MITF and MC1R ), were detected by gene annotation analysis. Finally, to fine-map the genomic region for the two-end black (TEB) coat color phenotype in Jianli pigs, we performed three signature selection methods between the TEB coat color and no-TEB coat color pig breeds. The current study, further confirmed that the EDNRB gene is a candidate gene for TEB color phenotype found in Chinese pigs, including Jinhua pigs, and the haplotype harboring 25 SNPs in the EDNRB gene may promote the formation of TEB coat color. Further ATAC-seq and luciferase reporter assays of these regions suggest that the 25-SNPs region was a strong candidate causative mutation that regulates the TEB coat color phenotype by altering enhancer function. Conclusion Our results advanced the understanding of the genetic mechanism behind artificial selection, and provided further resources for the protection and breeding improvement of Jianli pigs.

Glaesserella parasuis (GPS) can cause severe systemic inflammation in pigs, resulting in huge economic losses to the pig industry. At present, no effective method is available for the prevention and control of GPS infection. Molecular breeding for disease resistance is imminent, but disease-resistance genes have not been identified. To study the mechanism of systemic acute inflammation caused by GPS, we established three in vitro infection models (3D4/21 cells, PK15 cells, and PAVEC cells) according to its infection path. There was no significant difference in apoptosis among the three kinds of cells after 12 h of continuous GPS stimulation, while inflammatory factors were significantly upregulated. Subsequent transcriptome analysis revealed 1969, 1207, and 3564 differentially expressed genes (DEGs) in 3D4/21 cells, PK15 cells, and PAVEC cells, respectively, after GPS infection. Many of the DEGs were predicted to be associated with inflammatory responses (C3, CD44, etc.); cell proliferation, growth and apoptosis; gene expression; and protein phosphorylation. Key signaling pathways, including S100 family signaling, bacteria and virus recognition, and pathogen-induced cytokine storm signaling, were enriched based on Ingenuity Pathway Analysis (IPA). Furthermore, a total of three putative transmembrane receptors and two putative G-protein-coupled receptors, namely F3, ICAM1, PLAUR, ACKR3, and GPRC5A, were identified by IPA among the three types of cells. ACKR3 and GPRC5A play pivotal roles in bacterial adhesion, invasion, host immune response and inflammatory response through the S100 family signaling pathway. Our findings provide new insights into the pathological mechanisms underlying systemic inflammation caused by GPS infection in pigs, and they lay a foundation for further research on disease-resistance breeding to GPS.

Context:Adrenocortical carcinoma (ACC) is rare and have high rates of recurrence and mortality. The role of adjuvant radiation therapy (RT) in localized ACC was controversial. Methods:We conducted a retrospective study in our center between 2015 and 2021 to evaluate the efficacy and safety of adjuvant RT in localized ACC. Overall survival (OS) and disease-free survival (DFS) were estimated using the Kaplan-Meier method. Cox proportional hazards regression models were used to estimate the independent risk factors. Adverse events associated with RT were documented according to the toxicity criteria of the radiation therapy oncology group (RTOG) and the common terminology criteria for adverse events (CTCAE v5.0). Results:Of 105 patients with localized ACC, 46 (43.8%) received adjuvant RT after surgery. The median radiation dose was 45.0Gy (range:30.0-50.4) and median follow up time was 36.5 (IQR: 19.7-51.8) months. In comparison to the no adjuvant RT group, patients with adjuvant RT had better 3-year OS (87.9% vs 79.5%, P=0.039), especially for patients with ENSAT I/II stage (P=0.004). Adjuvant RT also improved the median DFS time from 16.5months (95%CI, 12.0-20.9) to 34.6months (95%CI, 16.1-53.0). Toxicity of RT was generally mild and moderate with six grade 3 events. Conclusions:Postoperative adjuvant RT significantly improved OS and DFS compared with the use of surgery alone in resected ACC patients. Although this retrospective study on RT in localized ACC indicates that RT is effective in ACC, its findings need to be prospectively confirmed.

Intramuscular fat (IMF) content plays a crucial role in determining pork quality. Recent studies have highlighted transcriptional mechanisms controlling adipogenesis in porcine IMF. However, the changes in chromatin accessibility during adipogenic differentiation are still not well understood. In this study, we performed the assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq) and transcriptome sequencing (RNA-Seq) analyses on porcine intramuscular preadipocytes to explore their adipogenic differentiation into mature adipocytes. We identified a total of 56,374 differentially accessible chromatin peaks and 4226 differentially expressed genes at day 0 and day 4 during adipogenic differentiation. A combined analysis of the ATAC-seq and RNA-seq data revealed that 1750 genes exhibited both differential chromatin accessibility and differential RNA expression during this process, including selenium-binding protein 1 (SELENBP1), PLIN1, ADIPOQ, and FASN. Furthermore, we found that vitamin D receptor (VDR) could bind to the promoter region of the SELENBP1 gene, activate SELENBP1 transcription, and ultimately promote lipid accumulation during adipogenic differentiation. This study provides a detailed overview of chromatin accessibility and gene expression changes during the adipogenic differentiation of porcine intramuscular preadipocytes. Moreover, we propose a novel regulatory mechanism involving the VDR-SELENBP1 signaling axis in adipogenic differentiation.

OBJECTIVE:Mineralocorticoid receptor antagonists are the recommended medical therapy for bilateral primary aldosteronism (BPA). Patients with BPA have higher risk of cardiocerebrovascular disease (CCVD) than those with essential hypertension. There is no consensus on the criteria to assess the effectiveness of medical therapy for BPA. This study aimed to investigate the incidence of and risk factors for CCVD after medical therapy of BPA. METHODS:We conducted a retrospective cohort study including 240 patients with BPA treated with mineralocorticoid receptor antagonists. The posttreatment plasma renin activity (PRA) was defined as unsuppressed (PRA, ≥1 ng/mL/h); otherwise, it was defined as suppressed. We analyzed the association of posttreatment PRA status with CCVD outcomes. RESULTS:Of patients with BPA, 7.1% (17/240) developed CCVD at a median follow-up of 5.0 (range, 2.96-7.66) years. Moreover, 57.1% of patients had a PRA of ≥1 ng/mL/h after treatment. Patients with a PRA of <1 ng/mL/h had a higher incidence of CCVD (12.6% vs 2.9%, P < .05) and were at higher risk than those with a PRA of ≥1 ng/mL/h (hazard ratio, 4.50 [95% CI, 1.47-13.83; P < .05]; adjusted hazard ratio, 3.98 [95% CI, 1.22-13.02; P < .05]). CONCLUSION:Patients with BPA who receive pharmacologic treatment have a high incidence of CCVD. PRA may be an indicator that mineralocorticoids are being adequately antagonized.

DNA methylation plays a critical role in regulating gene expression during testicular development. However, few studies report on candidate genes related to the DNA methylation regulation of porcine testicular development. This study examined the differentially expressed genes (DEGs) and their methylation levels in testicular tissues from pigs at 60 days of age (60 d) and 180 days of age (180 d) using RNA-Seq and whole genome bisulfite sequencing (WGBS). It was determined that DNA methylation primarily occurs in the cytosine–guanine (CG) context, and the analysis identified 106,282 differentially methylated regions (DMRs) corresponding to 12,385 differentially methylated genes (DMGs). Further integrated analysis of RNA-Seq and WGBS data revealed 1083 DMGs negatively correlated with the expression of DEGs. GO analysis showed that these genes were significantly enriched in spermatogenesis, germ cell development, and spermatid differentiation. The screening of enriched genes revealed that hyper-methylation repressed ADAM30, ADAM3A, DPY19L2, H2BC1, MAK, RPL10L, SPATA16, and YBX2, while hypo-methylation elevated CACNA1I, CADM1, CTNNB1, JAM2, and PAFAH1B3 expression. Additionally, the methylation status of the key genes ADAM3A, ADAM30, YBX2, JAM2, PAFAH1B3, and CTNNB1 was detected by bisulfite sequencing PCR (BSP). This study offers insights into the epigenetic regulation mechanisms underlying porcine testicular development.

Context Cushing syndrome (CS) is a severe endocrine disease characterized by excessive secretion of cortisol with multiple metabolic disorders. While gut microbial dysbiosis plays a vital role in metabolic disorders, the role of gut microbiota in CS remains unclear.Objective The objective of this work is to examine the alteration of gut microbiota in patients with CS.Methods We performed shotgun metagenomic sequencing of fecal samples from 78 patients with CS and 78 healthy controls matched for age and body mass index. Furthermore, we verify the cortisol degradation capacity of Ruminococcus gnavus in vitro and identify the potential metabolite by LC-MC/MS.Results We observed significant differences in microbial composition between CS and controls in both sexes, with CS showing reduced Bacteroidetes (Bacteroides vulgatus) and elevated Firmicutes (Erysipelotrichaceae_bacterium_6_1_45) and Proteobacteria (Enterobacter cloacae). Despite distinct causes of hypercortisolism in ACTH-dependent and ACTH-independent CS, we found no significant differences in metabolic profiles or gut microbiota between the 2 subgroups. Furthermore, we identified a group of gut species, including R. gnavus, that were positively correlated with cortisol levels in CS. These bacteria were found to harbor cortisol-degrading desAB genes and were consistently enriched in CS. Moreover, we demonstrated the efficient capacity of R. gnavus to degrade cortisol to 11-oxygenated androgens in vitro.Conclusion This study provides evidence of gut microbial dysbiosis in patients with CS and identifies a group of CS-enriched bacteria capable of degrading cortisol. These findings highlight the potential role of gut microbiota in regulating host steroid hormone levels, and consequently host health.

ContextThe prevalence of unilateral primary aldosteronism (UPA) with cortisol co-secretion varies geographically.ObjectiveTo investigate the prevalence and clinical characteristics of UPA with cortisol co-secretion in a Chinese population.DesignRetrospective cohort study.MethodsWe recruited 580 patients with UPA who underwent cosyntropin stimulation test (CST) after the 1-mg dexamethasone suppression test (DST) and retrospectively analyzed the clinical characteristics and postoperative outcomes of UPA with and without cortisol co-secretion.ResultsUPA with cortisol co-secretion (1 mg DST>1.8 ug/dL) was identified in 65 of 580 (11.2%) patients. These patients were characterized by older age, longer duration of hypertension, higher concentration of plasma aldosterone and midnight cortisol, lower adrenocorticotropic hormone (ACTH) and dehydroepiandrosterone sulfate (DHEAS), larger tumor diameter, and more history of diabetes mellitus. Cortisol and aldosterone levels were higher and DHEAS level was lower in UPA with cortisol co-secretion at 0–120 min after CST. Among 342 UPA patients with KCNJ5 gene sequencing and follow-up results, the complete clinical success rate was lower in UPA with cortisol co-secretion (33.3% vs. 56.4%, P<0.05); the complete biochemical success rate and KCNJ5 mutation did not differ between the two groups. Age, tumor size, and ACTH were independent predictors of UPA with cortisol co-secretion. Sex, BMI, duration of hypertension, KCNJ5 mutation, and cortisol co-secretion were independent predictors for complete clinical success in UPA after surgery.ConclusionsUPA with cortisol co-secretion is not uncommon in China, but the clinical features were distinctly different from those without co-secretion. Cortisol co-secretion is an independent risk factor for incomplete clinical success after surgery in UPA.