Search Results (5,132)

Diabetes mellitus (DM) is a chronic metabolic disease characterized by hyperglycemia, and the diaphragm—the primary respiratory muscle—is adversely affected by this diabetic process. The aim of this study is to investigate the effects of zinc sulfate (ZnSO₄) treatment on diaphragm muscle contractile dynamics, calcium homeostasis, apoptosis, and fibrosis in an 18-month-old female Type 2 diabetic rat model. Thirty-two 18-month-old female Wistar rats were randomly divided into four groups: Control (CON), CON + ZnSO₄, Diabetes Mellitus (DM), and DM + ZnSO₄. The DM model was induced by a high-fat diet and administration of 30 mg/kg streptozotocin (STZ); after the disease was confirmed, ZnSO₄ was administered intraperitoneally at a daily dose of 10 mg/kg to the treatment groups. The mechanical functions of the diaphragm muscle were evaluated using a post-rest potentiation protocol in an isolated organ bath; qPCR analyses (Caspase-3, TGF-β1, SERCA 2a) were performed to investigate cellular apoptosis, fibrosis, and calcium regulation. Compared with the CON group, the DM group exhibited a severe ~90% reduction in diaphragmatic contraction force (CF) and a ~97% decline in maximal contraction/relaxation velocities (±dF/dt_max) (p < 0.0001), which strongly correlated with a 30% suppression of SERCA2a gene expression (p < 0.01). Concomitantly, apoptotic Caspase-3 (~2.6-fold) and profibrotic TGF-β1 (~3.1-fold) mRNA levels were significantly elevated (p < 0.0001). In the DM + ZnSO₄ group, daily zinc treatment (10 mg/kg/day, i.p. for 6 weeks, initiated 4 weeks after diabetes confirmation) did not reverse the elevated Caspase-3 and TGF-β1 expressions (p > 0.05). However, SERCA2a expression was fully preserved back to control levels (p < 0.05 vs. DM), leading to a substantial ~3-fold improvement in CF and velocities (p < 0.05 to p < 0.0001 vs. DM). On the other hand, the healthy CON + ZnSO₄ group exhibited a physiological slowing of contractility (~53% decrease in CF), without histological damage, likely due to a competitive antagonism between excess divalent zinc (Zn²⁺) and calcium (Ca²⁺) on myofilaments. Although zinc cannot reverse the structural apoptotic and fibrotic remodeling in the aged diabetic diaphragm, it successfully rescues functional contractility by preserving SERCA2a transcriptional expression. Full article

Background: Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease driven by aberrant extracellular matrix deposition, with the activity of transforming growth factor-beta 1 (TGF-β1) orchestrating fibrogenesis. Current therapies are limited by severe adverse effects, highlighting the unmet need for safer treatments. In this study, the therapeutic potential of chlorogenic acid (CGA) in a murine model of pulmonary fibrosis was evaluated. Methods: Pathology was induced on day 0 via subcutaneous osmotic minipumps delivering bleomycin (BLM) for one week, followed by pump removal on day 10. Therapeutic interventions with intragastric CGA (60 mg/kg) were administered daily from days 14 to 20. Lung tissue samples obtained on day 21 were analyzed via histological, immunohistochemical, and RT–PCR methods. Results: Histological analysis via H&E and Masson’s trichrome staining revealed that CGA treatment significantly attenuated alveolar thickening, restored the alveolar space, and reduced the total cell density and Ashcroft fibrosis score. Furthermore, CGA markedly decreased collagen deposition. Quantitative RT–PCR and immunohistochemical assays revealed that CGA effectively decreased the expression of Col1a1, TGF-β1, and the myofibroblast marker alpha-smooth muscle actin (α-SMA). Conclusions: CGA exerts important therapeutic effects by decreasing the expression of TGF-β1 and the activation of myofibroblasts, indicating that this natural polyphenol is a promising candidate for mitigating the progression of pulmonary fibrosis. Full article

A 56-day feeding trial was designed to investigate the effects of alginate oligosaccharide (AOS) on the growth, immune response, antioxidant activity and apoptosis pathways of largemouth bass (Micropterus salmoides). We formulated six isonitrogenous and isoenergetic diets with different concentrations of AOS (0% (control), 0.05%, 0.1%, 0.15%, 0.2% and 0.25%). The results showed that the WGR of the AOS0.15–0.2 groups were markedly increased, and the FBW and SGR of the AOS0.1–0.2 groups were also markedly boosted. In addition, no significant differences were observed in FCR, SR and FI in the treatment groups. According to SGR and WG second-degree polynomial regression analysis, the optimum AOS addition level for juvenile largemouth bass was 0.14–0.15%. On the other hand, no notable differences were observed in crude protein, moisture, crude lipid or crude ash content between groups, and no notable differences were also observed in the levels of AST and ALT in the plasma between all groups. Additionally, ALP activities were considerably higher in the AOS0.15–0.25 groups. In terms of intestinal digestion and absorption function, AOS0.1 group and AOS0.15 group significantly increased the intestinal amylase and lipase activities, and A0S0.1–0.2 groups significantly increased the intestinal trypsin activities, while proper dietary supplementation with AOS significantly improved villus muscular thickness, villus height, and villus width. Furthermore, proper dietary supplementation with AOS significantly up-regulated the mRNA levels of occ, clau, C6A6, C7A5, C7A8B, C6A14 and pept1 in the intestine. No significant differences were observed in the mRNA levels of C7A6, C7A1A and C7A10A between all groups. With respect to the antioxidant and immune functions of the intestine, the analysis revealed no remarkable differences between the groups concerning SOD, GPX activity or T-AOC content in the intestine. However, a significant increase in CAT activity of the intestine was observed in the AOS0.05–0.15 groups, and MDA levels were lower in all AOS-added groups. Apart from the above, AOS0.15–0.25 groups significantly reduced intestinal TNF-α concentration. No notable differences were observed in the intestinal contents of TGF-β, IL-10 and IL-6 between all groups. Additionally, proper dietary supplementation with AOS could improve antioxidant effects and inhibit inflammation by regulating the gene expressions of the related-Nrf2 and NF-κB signaling pathway, including nrf2, keap1, Mn-sod, gpx, nf-κb, il-10 and tgf-β. There was no significant difference in the mRNA levels of cat, fox, il-8 and tnf-α. With respect to cell apoptosis in the intestine, TUNEL assay results showed that green positive cells were significantly lower in the AOS0.05–0.2 groups than the AOS0 group. Additionally, proper dietary supplementation with AOS could inhibit cell apoptosis by regulating the mRNA levels of bxl-xl, caspase 3, caspase 8, caspase 9 and bcl-2. However, there was no significant effect on the level of bax mRNA in any of the treatment groups. In summary, proper dietary supplementation with AOS exerted positive effects on growth, intestinal digestion and absorption function, immune antioxidant responses, and apoptosis pathways to a certain extent. Full article

Breast cancer relapse in bone is a significant clinical problem that is experienced in ~70–80% of patients with late-stage breast cancer. This condition commonly occurs 5–10+ years following surgical removal of the primary tumour. The long latency seen prior to relapse in bone is a result of tumour cell dormancy. Once disseminated to the bone, tumour cell interaction with the bone metastatic niche (endosteal niche and endovascular cells) maintains cells in a dormant state until changes to the local environment activate the niche to support outgrowth. Amassing evidence suggests that cytokines are key regulators of the bone metastatic niche, controlling bone homing and metastatic outgrowth. Pro-inflammatory cytokines, including IL-1β, IL-6, IL-8, TGFβ and RANKL, play crucial roles in attracting tumour cells to bone. Furthermore, these cytokines act in conjunction with IFN, VEGF, TGF, PTHrP, FGF, OPG and various chemokines to regulate expansion of the niche, facilitating tumour cell escape from dormancy and promoting the “vicious cycle of bone metastasis”. Here, we review the current literature to provide an up-to-date understanding of how interactions between cytokine signalling cascades regulate the bone metastatic niches to promote homing, dormancy or metastatic outgrowth of breast cancers. Because breast cancers are predominantly osteolytic, this review focuses on dormancy and metastatic outgrowth associated with lytic disease in addition to current advances in novel therapeutics aimed at preventing this condition through targeting dormant cells. Full article

Background: While live-cell therapies face significant translational hurdles, cell-free secretomes derived from fetal progenitor cells offer a unique, scalable approach natively programmed for scarless tissue repair. Methods: This GLP-compliant study evaluated the therapeutic efficacy spectrum of an off-the-shelf, clinical-grade, intratracheal lyophilized FE002-Lu lung fibroblast conditioned medium (LFCM) across five controlled Wistar rat models of induced lung injury: bleomycin (5 mg/kg), asbestos (100 µg/rat), hyperoxia (100% oxygen exposure), silica (30 mg/rat), and lipopolysaccharide (LPS, 4 mg/kg). Following lung injury induction and symptom onset, symptomatic rats were randomized to receive intratracheal LFCM (low, mid, or high dose) or a vehicle control every 4 days for 28 days. Results: The intervention demonstrated an exceptional safety profile, maintaining 100% survival with no severe procedural toxicity across all cohorts. Across all study arms, LFCM effectively attenuated pulmonary inflammation, reducing pro-inflammatory markers (IL-1β, IL-6, TNF-α, and CINC-1) in bronchoalveolar lavage fluid. Concurrently, high LFCM doses consistently elevated the anti-inflammatory cytokine IL-10. Within the lung tissue, mid- and high doses significantly reduced key pro-fibrotic drivers, including TGF-β1, TIMP-1, WISP-1, and hydroxyproline. Treatments consistently decreased α-SMA expression and pro-fibrotic gene mRNA levels, mitigating pulmonary myofibroblast activation. This correlated with reduced Ashcroft scores and collagen deposition, thereby preserving lung architecture. Notably, the FE002-Lu LFCM treatment exerted a biphasic regulation of extracellular matrix turnover: it elevated Cathepsin-D and MMP-12 in the bleomycin arm to actively clear newly deposited fibrotic debris, while reducing these markers in the hyperoxia and LPS arms to prevent acute collateral degradation of the native lung matrix. In both particulate models (asbestos and silica), the LFCM mid-dose established an optimal therapeutic threshold, avoiding the localized secretome saturation and pro-fibrotic exacerbation occasionally observed at higher doses. Conclusions: Lyophilized FE002-Lu LFCM acts as a potent, pleiotropic biologic that effectively resolves acute pulmonary inflammation and arrests progressive fibrotic remodeling across multiple distinct in vivo models. By overcoming the cold-chain and delivery limitations inherent to pulmonary live-cell therapies, this stable, cell-free secretome represents a highly scalable, “off-the-shelf” candidate poised for non-invasive, aerosolized clinical translation. Full article

Reproductive efficiency determines the economic benefits of the sheep industry, yet the molecular mechanisms underlying prolificacy remain incompletely understood. To investigate transcriptomic differences associated with sheep prolificacy, we performed Oxford Nanopore Technologies (ONT) full-length transcriptome sequencing on ovarian tissues collected during the estrous phase from high-prolificacy Small-tailed Han sheep and comparatively lower-prolificacy Wadi sheep (n = 3 biological replicates per group), with an average sequencing depth of approximately 6.3 Gb per sample. With screening thresholds of |log2FoldChange| > 1 and p < 0.05, transcriptomic analysis identified 457 differentially expressed genes (DEGs; 207 upregulated, 250 downregulated) and 1033 differentially expressed transcripts (DETs). In total, 55% of DETs exhibited expression changes independent of overall gene abundance, highlighting the potential role of alternative splicing (AS)-mediated post-transcriptional regulation. We detected 55,478 AS events and screened 96 significant differential alternative splicing (DAS) events (|∆PSI| > 0.1, p < 0.05) across 78 differentially spliced genes (DSGs). Functional enrichment showed DEGs were primarily associated with reproduction pathways (e.g., TGF-β, MAPK, and ovarian steroidogenesis), whereas DSGs were enriched in p53 signaling and ribosome pathways. Protein–protein interaction network analysis highlighted highly connected candidate genes, including INHBA, CYP19, TNFAIP6, TK1, RRM2, BIRC5, BCL2, ISG15, PCLAF, and MX1, potentially involved in follicular development and reproductive signaling. The results of this study enrich the full-length transcriptomic resources for Small-tailed Han sheep and Wadi sheep, and provide candidate genes and transcriptomic resources for further functional investigation of sheep prolificacy. Full article

Urban particulate matter (UPM) is a major environmental pollutant known to induce systemic inflammation. While its respiratory toxicity is well-documented, the potential vulnerability of the female reproductive system to inhaled UPM remains a critical concern. In this study, we evaluated female reproductive outcomes in a murine model following repeated short-term intratracheal instillation of UPM (40–200 mg/kg) for a month. Our results demonstrate that while UPM exposure induced robust, dose-dependent pulmonary inflammation, characterized by immune cell infiltration (CD45+, F4/80+, and Ly6G+) and upregulation of pro-inflammatory (Il-6, Il-1β, and Tnfα) and fibrotic (Col1a1 and Tgfβ1) markers, no apparent adverse effects were observed in the reproductive endpoints examined. No significant alterations were observed in maternal pregnancy outcomes, including oocyte recovery rates, fertilization competence, gestation duration, litter size, and postnatal development of pups. Furthermore, the mRNA expression of inflammatory markers in the ovaries and uteri was not detectable. Direct in vitro exposure of two-cell embryos to UPM did not interfere with preimplantation embryo development. Although benzanthracene slightly reduced the blastocyst formation rate, UPM and polycyclic aromatic hydrocarbons, major toxic components of UPM, did not severely compromise preimplantation embryo development in vitro. In conclusion, these findings suggest that marked UPM-induced pulmonary inflammation is not accompanied by substantial alterations in the female reproductive outcomes examined under short-term exposure conditions. Full article

Coronary artery disease is a leading cause of morbidity and mortality in modern medicine. In contrast, surgical myocardial revascularization via coronary artery bypass grafting (CABG) remains the gold standard of treatment for complex multivessel disease. The great saphenous vein remains the most frequently used conduit due to its availability and technical simplicity, but its long-term patency is significantly inferior to that of arterial grafts. The primary pathological process responsible for vein graft failure is intimal hyperplasia, which represents a complex response of the vascular wall to surgical trauma, vein arterialization, inflammation, and hemodynamic stress. This process is characterized by endothelial dysfunction, inflammatory cell activation, proliferation and migration of vascular smooth muscle cells, and extracellular matrix remodeling. Underpinning these alterations are numerous molecular pathways, including NF-κB, MAPK, PI3K/Akt, TGF-β, and mTOR signaling, as well as substantial contributions from oxidative stress, cytokines, growth factors, and microRNAs. Contemporary research indicates that the phenotypic transformation of vascular smooth muscle cells constitutes the central event in the development of intimal hyperplasia. Understanding the cellular and molecular mechanisms underlying this disease’s onset enables the development of novel therapeutic strategies to preserve long-term graft patency. This review paper aims to provide a systematic overview of current knowledge regarding the molecular and cellular mechanisms of intimal hyperplasia development in vein grafts following CABG. Full article

Sertoli cells (SCs) are essential for spermatogenesis and provide structural and nutritional support to germ cells in the Chinese Holstein cattle testis. Although microRNAs (miRNAs) are known to regulate SC function, the specific role of Bta-miR-146a in bovine SCs is unclear. This study investigated the mechanisms by which Bta-miR-146a regulates bovine immature SCs. Using molecular cloning, we constructed Bta-miR-146a overexpression and interference vectors and transfected them into SCs via lipofection. Quantitative real-time PCR (RT-qPCR), 5-ethynyl-2′-deoxyuridine (EdU) proliferation assays, Cell Counting Kit-8 (CCK-8) viability assays, and flow cytometry revealed that Bta-miR-146a overexpression inhibited SC proliferation and promoted apoptosis, whereas Bta-miR-146a inhibition increased proliferation and suppressed apoptosis. Dual-luciferase reporter assays confirmed that SMAD4 is a direct target of Bta-miR-146a; SMAD4 interference reduced SC proliferation and increased apoptosis, whereas overexpression had the opposite effect. Furthermore, activity of this gene modulates the TGFβ/MAPK signaling pathway; SMAD4 interference reduces the expression of TGFβ, TGF-βRII, DAXX, MAP3K5, P38, and MAX. These findings indicate that the Bta-miR-146a/SMAD4/TGFβ/MAPK axis is a key regulator of SC proliferation and apoptosis, offering insights into the molecular mechanisms underlying bovine spermatogenesis and potential targets for improving reproductive performance. Full article

Colorectal cancer (CRC) progression is shaped by dynamic interactions between tumor-intrinsic metabolic adaptations and immune remodeling within the tumor microenvironment. In CRC, the expression of SMAD7, a classical inhibitor of TGF-β1 signaling, is increased and has been associated with tumor-associated inflammatory responses and malignant progression. In this study, we investigated the potential role of SMAD7 in regulating glycolytic metabolism and macrophage phenotype in CRC. Knockdown of SMAD7 in CRC cell lines resulted in reduced glycolytic activity, as demonstrated by decreased extracellular acidification rate, basal glycolysis, and glycolytic capacity. These metabolic changes were associated with reduced expression of the basal and IL-6- and IL-22-induced glycolytic enzyme hexokinase 2 (HK2), while glucose uptake was increased. Similar reductions in HK2 expression were observed in patient-derived CRC organoids following SMAD7 inhibition, supporting the relevance of this pathway in human tumor-derived models. Functionally, SMAD7 knockdown reduced lactate production by CRC cells and diminished the ability of tumor cell-derived conditioned medium to induce the expression of macrophage-associated immunoregulatory markers, including CD163, CD206, and ARG1. The addition of exogenous lactate restored these effects, indicating that tumor-derived lactate contributes to SMAD7-dependent control of the expression of macrophage-associated immunoregulatory markers. Analysis of human CRC transcriptomic datasets revealed positive associations between SMAD7 expression and macrophage-related signatures, including profiles associated with immunoregulatory tumor-associated macrophages. Together, these findings support a potential role for SMAD7 in controlling tumor metabolism and macrophage-associated immunoregulatory markers in CRC. Full article

Non-small cell lung cancer (NSCLC) remains a leading cause of cancer-related mortality. Smoking is the primary etiological factor, but growing evidence suggests the involvement of estrogen in its development and progression, although its role remains unclear. This study explores: (i) the effects induced by estrogen, namely, 17β-estradiol (E2), alone or in combination with a mixture of inflammatory cytokines (Mix), in two human NSCLC cell lines, A549 and Calu1, and (ii) whether and how tumor cells can modulate the activation of normal lung fibroblasts. We found that E2 significantly enhances migration, invasion, and epithelial–mesenchymal transition in NSCLC cells, as well as their resistance to cisplatin, particularly in combination with Mix. Pharmacological inhibition of ERβ reversed the E2-induced effects, implicating ERβ in E2-mediated signaling. Furthermore, E2 increased autophagic flux and induced a shift toward secretory autophagy and the release of extracellular vesicles, which activated normal lung fibroblasts, as demonstrated by the increased expression of α-SMA, FAP, PDGFR-β, and PDPN. The clinical relevance of these data was supported by computational analyses revealing an elevated expression of the Mix gene signature, including TGF-β, IL-6, IL-8, CCXL-16, and ERβ, which was associated with shorter overall survival in NSCLC patients. This molecular profile was linked to the elevated expression of secretory autophagy genes and cancer-associated fibroblast markers. Validation in three large clinical cohorts (TCGA-LUNG, OAK and POPLAR) strengthens the clinical relevance of this E2-related pro-tumor axis while suggesting a promising therapeutic avenue for NSCLC patients. Full article

The Fontan circulation is associated with progressive multisystem complications, yet its underlying molecular mechanisms remain incompletely understood. We aimed to characterize systemic proteomic alterations and identify biological pathways associated with Fontan physiology, long-term adaptation, and clinically unfavorable phenotypes. Peripheral vein serum proteomics was performed using mass spectrometry in 48 Fontan patients and matched controls. Differential protein expression and pathway enrichment analyses were applied across the overall cohort and clinically defined subgroups according to Fontan duration and composite clinical risk phenotyping. More than one third of quantified proteins were differentially expressed, demonstrating extensive systemic proteomic remodeling. Extracellular matrix remodeling emerged as the dominant signature, characterized by fibrosis-associated proteins and enrichment of TGF-β-related profibrotic pathways. Prolonged Fontan duration and clinically unfavorable status were not associated with substantially amplified extracellular matrix signatures. Instead, long-term Fontan patients demonstrated increasing immune, complement, and endothelial activation, whereas high-risk patients showed metabolic and redox-related alterations. These findings indicate that extracellular matrix remodeling and profibrotic signaling are dominant features of the Fontan circulation, while increasing Fontan duration and clinical deterioration are associated with immune activation, endothelial stress, and metabolic dysregulation with oxidative stress. Full article

Background: Homeobox transcription factors regulate developmental programs, cellular plasticity, and tumor progression, yet the role of H2.0-like homeobox (HLX) in prostate cancer (PCa) remains poorly defined. We investigated the clinical significance of HLX and its relationship to the pro-metastatic transcription factor TBX2. Methods: Transcriptomic and clinical datasets from TCGA, MET500, and SU2C/PCF cohorts were analyzed to assess HLX expression, clinicopathologic associations, and its relationship with TBX2. Functional studies in human PCa cell lines included TBX2 gain- and loss-of-function, HLX knockdown, chromatin immunoprecipitation (ChIP), and expression analyses. Shared HLX- and TBX2-associated pathways were evaluated by Reactome enrichment analysis, and Hallmark Gene Set Enrichment Analysis compared castration-resistant prostate cancer (CRPC) bone metastases with high versus low HLX expression (GSE77930; n = 5/group). In vivo relevance was assessed in an orthotopic TBX2 dominant-negative PCa xenograft model. Results: Human PCa datasets showed that HLX expression was elevated in PCa versus normal prostate tissue and associated with higher Gleason grade, lymph node involvement, aggressive molecular subtypes, and shorter disease-free survival. HLX expression also positively correlated with TBX2 across human PCa cohorts. HLX- and TBX2-associated transcriptional programs converged on extracellular matrix organization, cell adhesion, NOTCH, and VEGF-MAPK signaling pathways. Furthermore, HLX-high CRPC bone metastases were enriched for epithelial–mesenchymal transition, NOTCH, TGF-β, inflammatory, angiogenic, hypoxic, and KRAS signaling pathways. Mechanistic studies showed that HLX knockdown suppressed extracellular matrix-associated genes and key NOTCH pathway components. ChIP demonstrated direct TBX2 binding to the HLX promoter, and genetic modulation of TBX2 expression established HLX as a downstream target of TBX2. Consistent with these findings, reduced HLX expression in orthotopic TBX2 dominant-negative xenografts was associated with loss of metastatic progression. Conclusions: HLX is a candidate biomarker of aggressive PCa and a direct transcriptional target of TBX2. These findings identify a previously unrecognized TBX2–HLX regulatory axis associated with metastatic transcriptional programs and aggressive disease in advanced PCa. Full article

Deoxynivalenol (DON) and patulin (PAT) are common mycotoxins in cereals and fruits, posing health risks for animals and human beings. In order to study their liver toxicity, 24 mice were randomly assigned to four groups, with six replicates in each group (one mouse per cage). The mice were intragastrically administered with DON, PAT, DON + PAT (DP), or without DON and PAT (the control group) for 28 days, respectively. The results showed that body weight gain was significantly reduced by all toxin treatments, compared with the control group, and the lowest body weight gain was observed in the DP group. Histopathology revealed that hepatocyte damage and inflammatory infiltration were more serious in the DP group, exhibiting the highest mRNA abundances of MyD88, IFN-γ, and JAK2. The severity of hepatocyte apoptosis induced in each group followed the order: DON > DP > PAT; the severity of oxidative stress was ranked as DP > DON > PAT. Transcriptomic analysis revealed that numerous differentially expressed genes were regulated by DP treatment, which were mainly enriched in the MAPK, JAK-STAT, and transforming growth factor (TGF)-β signaling pathways. In conclusion, individual exposure to DON or PAT triggered hepatic injury, and their co-exposure further exacerbated liver damage. Full article