Free fatty acids like palmitic acid (PA) are elevated in obesity and diabetes and dysregulate monocyte and macrophage functions, contributing to enhanced inflammation in these cardiometabolic diseases. Epigenetic mechanisms regulating enhancer functions play key roles in inflammatory gene expression, but their role in PA-induced monocyte/macrophage dysfunction is unknown. We found that PA treatment altered the epigenetic landscape of enhancers and super-enhancers (SEs) in human monocytes. Integration with RNA-seq data revealed that PA-induced enhancers/SEs correlated with PA-increased expression of inflammatory and immune response genes, while PA-inhibited enhancers correlated with downregulation of phagocytosis and efferocytosis genes. These genes were similarly regulated in macrophages from mouse models of diabetes and accelerated atherosclerosis, human atherosclerosis, and infectious agents. PA-regulated enhancers/SEs harbored SNPs associated with diabetes, obesity, and body mass index indicating disease relevance. We verified increased chromatin interactions between PA-regulated enhancers/SEs and inflammatory gene promoters and reduced interactions at efferocytosis genes. PA-induced gene expression was reduced by inhibitors of BRD4, and NF-κB. PA treatment inhibited phagocytosis and efferocytosis in human macrophages. Together, our findings demonstrate that PA-induced enhancer dynamics at key monocyte/macrophage enhancers/SEs regulate inflammatory and immune genes and responses. Targeting these PA-regulated epigenetic changes could provide novel therapeutic opportunities for cardiometabolic disorders.

Exposure to di-n-butyl phthalate (DBP) during embryo development or lactation has been linked to reproductive toxicity. The ten-eleven translocation (TET) protein family plays a role in various pathological processes; however, its involvement in reproductive dysfunction in offspring mice exposed to DBP during gestation remains sparsely reported. In this study, SPF C57BL/6 pregnant mice were intragastrically administered DBP at doses of 0.5, 5, and 75 mg/kg body weight, or corn oil as a control, from gestational days 5-19. Following weaning, the offspring mice were maintained on a standard diet for 5 weeks. Additionally, mono-n-butyl phthalate (MBP)-induced TM3 cells were utilized to explore the underlying mechanisms in vitro. The results showed that in utero exposure to DBP resulted in diminished sperm quality, testicular damage, decreased reproductive hormone levels, and reduced expression of testosterone synthesis proteins in male offspring mice. Moreover, DBP exposure influenced the expression of steroidogenic acute regulatory protein (StAR) via the cAMP/PKA signaling pathway, associated with luteinizing hormone receptor (LHR)-mediated suppression of testosterone synthesis. Notably, DBP exposure led to decreased expression of TET methylcytosine dioxygenase 2 (TET2) in the progeny, and overexpression or silencing of TET2 affected the levels of proteins involved in the LHR-mediated testosterone synthesis pathway. Further investigations revealed that TET2 downregulation inhibits testosterone synthesis through the LHR-mediated LH/cAMP/PKA/StAR signaling pathway, ultimately impairing reproductive function in DBP-exposed offspring mice during gestation. This study provides a novel perspective for identifying molecular markers that may be more sensitive indicators of male reproductive damage from an epigenetic standpoint.

Fetal alcohol spectrum disorders (FASD) describe ethanol-induced developmental defects including craniofacial malformations. While ethanol-sensitive genetic mutations contribute to facial malformations, the impacted cellular mechanisms remain unknown. Signaling via bone morphogenetic protein (Bmp) is a key regulatory step of epithelial morphogenesis driving facial development, providing a possible ethanol-sensitive mechanism. We found that zebrafish carrying mutants for Bmp signaling components are ethanol-sensitive and affect anterior pharyngeal endoderm shape and gene expression, indicating that ethanol-induced malformations of the anterior pharyngeal endoderm cause facial malformations. By integrating FASD patient data, we provide the first evidence that variants of the human Bmp receptor gene BMPR1B associate with ethanol-related differences in jaw volume. Our results show that ethanol exposure disrupts proper morphogenesis of, and tissue interactions between, facial epithelia that mirror overall viscerocranial shape changes and are predictive for Bmp-ethanol associations in human jaw development. Our data provide a mechanistic paradigm linking ethanol to disrupted epithelial cell behaviors that underlie facial defects in FASD.

Changweiqing (CWQ) is a Chinese herbal formula for the treatment of the gastrointestinal tract diseases, but its role in the treatment of colorectal cancer (CRC) has not been clarified. This study aimed to explore the molecular mechanism of CWQ in CRC treatment through bioinformatics analysis and network pharmacology.

Diazinon is a commonly used organophosphate (OP) insecticide especially in developing countries for the control of insect pests, however, exposure to its toxic impact especially in humans and other non-target species remains an important public health concern. The study aimed to investigate the effect of epigallocatechin -3- gallate (EGCG), abundant in green tea plants on neurobehavioural, biochemical, and pathological changes in the brain of male Wistar rats following exposure to diazinon toxicity. Sixty adult male Wistar rats were acclimatized for seven days and subsequently randomly assigned into six treatment groups as follows: Group I: Control group (0.2 mL distilled water); Group II: Diazinon at 3 mg/kg (1% LD50); Group III: Diazinon (3 mg/kg) + EGCG (50 mg/kg, ~ 2% of LD50); Group IV: Diazinon (3 mg/kg) + EGCG (100 mg/kg, ~ 5% of LD50); Group V: EGCG (50 mg/kg) and Group VI: EGCG (100 mg/kg). All treatments were administered orally once daily for 14 days. Neurobehavioural studies, biomarkers of oxidative stress, histology, immunohistochemistry, and quantitative polymerase chain reaction (RT qPCR) were performed. Diazinon alone impaired recognition memory, increased oxidative stress markers and altered antioxidant defense in the brain. It upregulated TNF-α and IL-6 genes and repressed GPx 4 gene expressions. It was also associated with increased GFAP, Tau, and α-SN immunoreactivity. Microscopic examination revealed loss of Purkinje and hippocampal cells in brain. Co-treatment with EGCG however improved cognition, lowered oxidative stress markers, improved antioxidant status and suppressed TNF-α and IL-6. In conclusion, findings from this study demonstrated that EGCG offered protection against diazinon-induced neurotoxicity. Hence, natural sources of epigallocatechin -3- gallate such as fruits and vegetables could offer immense benefits by protecting against oxidative stress and inflammation in neurodegenerative disease conditions.Clinical trial number Not applicable.

Aloe vera is a popular medicinal plant in the cosmetic, pharmaceutical, and food industries. Acemannan (ACE), a β-(1,4)-acetylated mannan, is one of the bioactive compounds isolated from the A. vera gel. The pharmacological effects of ACE have been reported regarding digestive disease protection, antimicrobia, and prebiotic activity. Here, we used human HaCaT cells as a model to uncover the potential biological functions of ACE in keratinocytes. ACE increased cell growth in a concentration-dependent manner, and a higher incorporation of BrdU was detected in ACE-treated cells than in vehicle-treated cells, indicating ACE promotes cell proliferation. Furthermore, ACE concentration-dependently promoted cell migration in the wound scratch model. ACE regulated cell differentiation by transiently decreasing p63α expression, but increasing the expression of involucrin, loricrin, and transglutaminase 1 (TGase 1). These effects were non-additive to those induced by phorbol myristate acetate (PMA), but additive to epidermal growth factor (EGF), which are complete and incomplete differentiation agents of keratinocytes, respectively. Moreover, ACE activated EGF receptor (EGFR), protein kinase C (PKC), and protein kinase B (AKT/PKB). PKC inhibitor Ro320432 enhanced cell growth and migration, while EGFR inhibitor osimertinib blocked both responses. In summary, ACE is a potential therapeutic agent in wound healing. ACE activates PKC, leading to keratinocyte differentiation and activates EGFR, contributing to keratinocyte proliferation and migration.

Synovitis represents the initial pathological change in osteoarthritis and contributes to its progression. Resolvin D1 (RV-D1) is a novel and endogenous docosahexaenoic acid-derived lipid mediator, which regulates the duration and magnitude of inflammation by downregulating pro-inflammatory genes and mediators. However, the effects of RV-D1 on synovitis remain unknown. The aim of the present study was to investigate the anti-inflammatory effects of RV-D1 in human fibroblast-like synoviocytes (HFLSs) and the underlying mechanisms. The expression of the HFLS formyl peptide receptor 2 (ALX/FPR) was examined via immunocytochemical analysis. HFLSs were treated with 1 ng/mL recombinant human interleukin-1β (IL-1β) and RV-D1. The gene expression of interleukin-1β (IL1B), matrix metalloproteinase 3 (MMP3), and MMP13 was examined using real-time reverse transcription-polymerase chain reaction after treatment with IL-1β and RV-D1. The effect of RV-D1 on apoptosis was examined based on fluorescence intensity. Phosphorylation of p-38, extracellular signal-regulated kinase, c-Jun N-terminal kinase, nuclear factor kappa B (NF-κB), and AKT was analyzed via western blotting. ALX/FPR staining was observed on the cell surface. RV-D1 significantly suppressed the IL-1β-induced increase in gene and protein expression of IL-1β, MMP-3, and MMP-13. Pretreatment with 100 nM RV-D1 significantly increased the fluorescence intensity compared to that in the non-treatment group. Furthermore, pretreatment with RV-D1 significantly suppressed the phosphorylation of p-38, NF-κB, and AKT. Whereas WRW4, an antagonist of ALX/ FPR2, treatment weakened the effect of RV-D1, resulting in p-38, NF-κB, and AKT phosphorylation and the protein expression of MMP-13 at levels comparable to those in the IL-1β without RV-D1. In conclusion, RV-D1 suppressed IL-1β and MMP expression by inhibiting the phosphorylation of p-38, NF-κB, and AKT in inflammation in HFLSs. RV-D1 can be used to develop treatments for osteoarthritis and other inflammatory disorders.

Ovarian cancer therapy remains a challenge for human health, partly due to chemotherapy resistance. Understanding the molecular mechanisms underlying this resistance is crucial. Therefore, to identify genes involved in cisplatin resistance in ovarian cancer, RNA-seq analysis of A2780cp (cisplatin-resistant) and A2780 (cisplatin-sensitive) cell lines was performed, revealing 1-acylglycerol-3-phosphate O-acyltransferase 3 (AGPAT3) as a differentially expressed candidate gene. First, MTT analysis confirmed the drug resistance of A2780cp and the sensitivity of A2780 cell lines. Subsequent reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blotting analyses revealed elevated AGPAT3 and mTOR expression in A2780cp cells compared with A2780 cells. Additionally, western blotting showed increased p-mTOR (phospho-mTOR)/mTOR and p-S6K (phospho-S6K)/S6K ratios in A2780cp cells. The overexpression of AGPAT3 in A2780 cells led to increased p-mTOR/mTOR and p-S6K/S6K ratios and increased IC50 values, as shown by RT-qPCR, western blotting, and MTT analysis. Conversely, shRNA-mediated downregulation of AGPAT3 resulted in reduced p-mTOR/mTOR and p-S6K/S6K ratios. At the cellular level, AGPAT3 overexpression in A2780 cells increased survival rates, decreased apoptosis, and caused G2/M cell cycle arrest under cisplatin treatment, as detected by apoptosis assay, and cell cycle flow cytometry analysis. Overall, we conclude that AGPAT3 is involved in cisplatin resistance in A2780cp cells and propose that targeting this gene or its enzymatic product could help overcome drug resistance.

Calcific aortic valve disease (CAVD) is a progressive, age-related degenerative disease characterized by the accumulation of calcium deposits in the aortic valve. We aim to screen key genes associated with cellular senescence (CS) in CAVD.

Acute myocardial infarction (AMI) is a leading cause of heart failure, often accompanied by myocardial fibrosis (MF), characterized by excessive extracellular matrix accumulation. Endothelial-to-mesenchymal transition (EndMT) plays a key role in MF progression post-AMI. Neuregulin-1 (NRG-1), a growth factor with cardioprotective properties, has emerged as a potential therapeutic target. Tongxinluo (TXL), a traditional Chinese medicine, mitigates MF by upregulating NRG-1. This study elucidates the mechanisms underlying the protective effects of NRG-1 and TXL against MF following AMI. Left anterior descending artery ligation established a model for mice with AMI. Adeno-associated virus was used to modulate NRG-1 expression in the myocardium. Echocardiography assessed cardiac function, and histological staining was used to evaluate MF. Expression levels of markers for myofibroblasts (α-SMA, FSP-1) and endothelial cells (CD31, VE-cadherin) were analysed to investigate EndMT. The involvement of the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signalling pathway in NRG-1's protective mechanism was validated using biochemical methods. Tongxinluo was administered to mice with AMI via gavage for 4 weeks, and its effects on cardiac function, MF and EndMT were assessed. Overexpression of NRG-1 in mice with AMI ameliorated cardiac dysfunction and reduced interstitial and perivascular fibrosis, whereas NRG-1 deficiency exacerbated these effects. NRG-1 protected against EndMT, as evidenced by changes in myofibroblast and endothelial cell markers. The PI3K/AKT signalling pathway was involved in NRG-1's protective mechanism against MF. The administration of TXL to mice with AMI improved cardiac function and reduced MF by activating NRG-1. Furthermore, TXL inhibited EndMT post-AMI through the NRG-1/PI3K/AKT pathway. NRG-1 and TXL protect against MF post-AMI by mitigating EndMT through the PI3K/AKT pathway. These findings suggest that targeting NRG-1 or using TXL may be promising therapeutic strategies for MF following AMI.

We previously demonstrated that the midnolin gene (MIDN) is a risk factor for Parkinson's disease (PD) in Yamagata and British cohorts, and that neurite outgrowth is abolished by MIDN knockout in PC12 cells. Therefore, drugs that upregulate MIDN may have neurotrophic effects. In this study, acetylcholine increased MIDN promoter activity and gene expression in a concentration-dependent manner in SH-SY5Y cells. These effects were suppressed by atropine and a Gq inhibitor, YM254890, indicating that muscarinic receptor/Gq signaling is required for the induction of MIDN by acetylcholine. Our findings suggest that drugs that upregulate MIDN may have therapeutic potential for PD.

Epigenetic aging biomarkers are used for evaluating morbidity and mortality, monitoring therapies, and direct-to-consumer testing. However, the influence of environmental exposures on epigenetic age acceleration (EAA), also known as epigenetic age deviation, has not been systematically evaluated. In this systematic review, we synthesized findings from human epidemiologic studies on chemical and climatic environmental exposures, particularly air pollution, chemicals, metals, climate, and cigarette smoke, and EAA. A total of 102 studies analyzing epigenetic data from over 180,000 subjects were evaluated. Overall, studies in each exposure category frequently included adult participants, used a variety of epigenetic clocks, analyzed whole blood samples, and had a low risk of bias. Exposure to air pollution (15/19 of studies; 79%), cigarette smoke (53/66; 80%), and synthetic and occupational chemicals (5/8; 63%) were notably associated with increased EAA. Results for essential and non-essential metal exposure were more equivocal: 7/13 studies (54%) reported increased EAA. One study reported increased EAA with greater temperature exposure. In summary, we identified environmental exposures, such as air pollution and cigarette smoke, that were strongly associated with increased EAA. Further research is needed with larger and more diverse samples and high-quality exposure assessment.

Accumulating evidence indicates that drug addiction may lead to adaptive behavioral changes in offspring, potentially due to epigenetic modifications in parental germline. However, the underlying mechanisms remain inadequately understood. In this study, we show that paternal heroin self-administration (SA) increased heroin-seeking behavior in the F1 generation, when compared with offspring sired by yoke-infused control males, indicating cross-generational impact of paternal voluntary heroin seeking behavior. Notably, the increase of heroin seeking behavior in offspring was replicated by zygotic microinjection of sperm RNAs derived from sperm of heroin-SA-experienced rats. Analysis of non-coding RNAs in spermatozoa revealed coordinated changes in miRNA content between the nucleus accumbens and spermatozoa. We validated that restoration of miR-19b downregulation in sperm RNA from self-administration-experienced rats, in parallel with its overexpression in the nucleus accumbens of F1 offspring sired by heroin-SA-experienced fathers, reversed the increased heroin SA observed in these F1 offspring. Taken together, our findings suggest in rats that paternal heroin self-administration induces epigenetic changes in both brain and sperm miRNA, with miR-19b downregulation playing a critical role in mediating the epigenetic inheritance of increased heroin self-administration behavior in the F1 generation.

Lipid metabolic reprogramming is increasingly recognized as a hallmark of endocrine resistance in estrogen receptor-positive (ER+) breast cancer. In this study, we investigated alterations in lipid metabolism in ER + breast cancer cell lines with acquired resistance to common endocrine therapies and evaluated the efficacy of a clinically relevant fatty acid synthase (FASN) inhibitor.

Abscisic acid (ABA) is a crucial endogenous hormone in plants, regulating a myriad of processes integral to plant growth and development. However, there has been no reported impact on the production of aromatic rice following the application of exogenous ABA. Aromatic rice not only possesses intense aroma but also boasts higher nutritional value. 2-Acetyl-1-pyrroline (2-AP) is the primary compound responsible for the distinctive aroma of aromatic rice. A two-year field experiment was conducted to explore the impact of exogenous ABA application on the biosynthesis and accumulation of 2-AP, as well as the physiological characteristics, yield, and quality of aromatic rice varieties.

Despite being one of the primary and most effective treatments for advanced stages of lung cancer, chemotherapy drugs like carboplatin have limitations due to their adverse side effects and the development of drug resistance in lung cancer cells. However, recent studies have shown promising results in using small interfering RNAs (siRNAs) as a therapeutic agent for cancer treatment. Hence, this study aimed to investigate the potential of combining siRNA-DLGAP1-AS2 with carboplatin in human lung cancer cell lines. The viability of the cells was assessed using the MTT assay, and apoptosis induction was examined through Annexin V/Pi staining. Additionally, the effect of the combination on cell cycle arrest and colony formation of lung cancer cells was studied. Furthermore, the expression of Bax, Bcl-2, MMP-2, MMP-9, GCLC, and CD44 was evaluated. Our functional analysis revealed that inhibiting the expression of DLGAP1-AS2 increased the sensitivity of lung cancer cells to carboplatin. Moreover, our study demonstrated that the combination of DLGAP1-AS2 inhibition through siRNA-DLGAP1-AS2 transfection and carboplatin treatment had a tumor-suppressive function, inhibiting the progression and proliferation of A549 lung cancer cells. Therefore, it can be concluded that targeting DLGAP1-AS2 using specific siRNA in combination with carboplatin chemotherapy holds promise as a valuable therapeutic approach for lung cancer.

Disrupted pH homeostasis can precipitate cell death and represents a viable therapeutic target in oncological interventions. Here, we utilize mass spectrometry-based drug analysis, transcriptomic screens, and lipid metabolomics to explore the metabolic mechanisms underlying pH-dependent cell death. We reveal CYP51A1, a gene involved in cholesterol synthesis, as a key suppressor of alkalization-induced cell death in pancreatic cancer cells. Inducing intracellular alkalization by the small molecule JTC801 leads to a decrease in endoplasmic reticulum cholesterol levels, subsequently activating SREBF2, a transcription factor responsible for controlling the expression of genes involved in cholesterol biosynthesis. Specifically, SREBF2-driven upregulation of CYP51A1 prevents cholesterol accumulation within lysosomes, leading to TMEM175-dependent lysosomal proton efflux, ultimately resulting in the inhibition of cell death. In animal models, including xenografts, syngeneic orthotopic, and patient-derived models, the genetic or pharmacological inhibition of CYP51A1 enhances the effectiveness of JTC801 in suppressing pancreatic tumors. These findings demonstrate a role of the CYP51A1-dependent lysosomal pathway in inhibiting alkalization-induced cell death and highlight its potential as a targetable vulnerability in pancreatic cancer.

Cyclic diguanylate (c-di-GMP) is a central biofilm regulator in Pseudomonas aeruginosa, where increased intracellular levels promote biofilm formation and antibiotic tolerance. Targeting the c-di-GMP network may be a promising anti-biofilm approach, but most strategies studied so far aimed at eliminating surface-attached biofilms, while in vivo P. aeruginosa biofilms often occur as suspended aggregates. Here, the expression profile of c-di-GMP metabolism-related genes was analysed among 32 P. aeruginosa strains grown as aggregates in synthetic cystic fibrosis sputum. The diguanylate cyclase SiaD proved essential for auto-aggregation under in vivo-like conditions. Virtual screening predicted a high binding affinity of echinacoside towards the active site of SiaD. Echinacoside reduced c-di-GMP levels and aggregate sizes and potentiated tobramycin activity against aggregates in >80% of strains tested. This synergism was also observed in P. aeruginosa-infected 3-D alveolar epithelial cells and murine lungs, demonstrating echinacoside's potential as an adjunctive therapy for recalcitrant P. aeruginosa infections.

Natural killer/T-cell lymphoma (NKTCL) is a highly aggressive non-Hodgkin lymphoma. Relapsed/refractory (R/R) NKTCL patients have dismal prognosis and lack effective treatments, novel therapeutics are urgently needed. Here we found interleukin-2-inducible T-cell kinase (ITK) expression was elevated in NKTCL cells and patient tumors. And higher ITK expression was associated with worse clinical outcomes. In vitro ITK knockdown inhibited NKTCL cell growth, induced apoptosis, cell cycle arrest and impaired its colony-forming ability while ITK overexpression accelerated cell proliferation. In vivo ITK knockdown greatly impeded lymphoma growth in mouse model, indicating it as a potential therapeutic target. Mechanistically, ITK knockdown inhibited NKTCL cell growth by attenuating oncogenic NF-κB signaling, which is revealed by transcriptomic profiling and further validated by in vitro assays and in vivo NKTCL models. Additionally, we showed that ITK inhibitors could inhibit NKTCL cell proliferation, promote apoptosis and suppressed tumor progression in NKTCL cell line-derived xenograft (CDX) model. Furthermore, we established a patient-derived xenograft (PDX) model from a NKTCL patient refractory to prior anti-PD-1 and asparaginase containing therapy. The primary cells from this patient highly expressed ITK and were responsive to ITK inhibitor. And ITK inhibitor effectively repressed tumor progression in PDX model. Finally, we found ITK inhibition improved the response of NKTCL cell lines to chemotherapy and overcome chemotherapy resistance in primary cells. Collectively, our results demonstrated that ITK served as an oncogene in NKTCL and represented a novel therapeutic vulnerability to be targeted or in combination with chemotherapy drugs for this disease.

Heat stress (HS) poses a substantial challenge in the poultry sector, resulting in considerable economic losses as it negatively impacts the well-being and productivity of chickens. Dried plum (DP) is a rich source of minerals, vitamins, antioxidants, and phenolic compounds. Studies have indicated that DP offers various health advantages, including preserving the body's redox system, immune function, and calcium balance. In our previous study, DP supplementation improved overall growth performance and intestinal health metrics in heat-stressed broilers. Considering the beneficial effects of DP on health, we hypothesized that adding DP to the diet would mitigate the harmful impacts of heat stress in the liver of broiler chickens. Day-old unsexed broiler chicks (n = 72) were raised under standard conditions and randomly assigned to three treatment groups (n = 24/group): 1) Control, 2) heat stress with basal diet (HS), and 3) heat stress with supplement (DP). During the finisher stage, the DP group received feed containing 2.5% DP during treatment, while the other groups were given a standard finisher diet. After 21 d, birds in the HS and DP groups were subjected to cyclic heat stress conditions for 3 wk. The heat stress conditions involved exposing the birds to a temperature of 33-35°C for 8 h during the daytime. In contrast, the birds in the Control group were raised under normal conditions with temperatures ranging from 22-24°C. DP supplementation significantly increased (P < 0.05) heat shock factor 1 (HSF1) expression in the liver compared to the Control group. DP supplementation significantly increased (P < 0.05) thioredoxin (TXN), peroxiredoxin (PRDX), insulin-like growth factor 1 (IGF1), and methyl-CpG binding domain (MBD4) expression in the DP group compared to the HS group. Fructose-1,6-bisphosphatase 1 (FPB1) expression was significantly decreased (P < 0.05) in the DP group compared to the HS group. Solute Carrier Family 3 Member 1 (SLC3A1), DNA methyltransferase 1 (DNMT1), DNA methyltransferase 3 alpha (DNMT3A), ten-eleven translocation methylcytosine dioxygenase 2 (Tet2), ten-eleven translocation methylcytosine dioxygenase (Tet3), and thymine DNA glycosylase (TDG) expression were significantly increased (P < 0.05) in the DP group compared to the other treatment groups. In conclusion, post-hatch DP supplementation lessened the negative effects of HS on broiler chickens by upregulating genes related to heat shock, antioxidants, growth, nutrient transporters, and epigenetics in the liver.