Skeletal muscle atrophy is a common complication of heart failure, with myocardial infarction (MI) being the primary cause. Yet, the mechanisms linking post-MI cardiac insufficiency to muscle atrophy have remained unclear. The molecular basis for the beneficial effects of exercise on exercise intolerance in MI patients also remains absent. Serum IL-27 levels were measured in 48 MI patients and correlated with cardiac injury markers. Along with this, a rat model of post-MI cardiac insufficiency was used to assess skeletal muscle mass, cross-sectional area (CSA) of muscle fibers, and the expression of atrophy-related (MAFbx, MuRF-1) and differentiation-related markers (MyoD, Myogenin). The impact of exercise on muscle atrophy, cardiac inflammation, and IL-27 expression was then evaluated, with a focus on macrophage polarization. In vitro, the effects of macrophage-derived IL-27 on L6 myotube metabolism were assessed, and the role of HIF-1α in macrophage-derived IL-27 secretion was examined using co-culture systems. Serum IL-27 level was significantly elevated in MI patients and that it was positively correlated with myocardial injury and cardiac insufficiency. In post-MI rats, skeletal muscle mass and CSA of muscle fibers were reduced. Meanwhile, the expression level of myogenic markers was downregulated, while that atrophy markers was upregulated. IL-27 treatment promoted catabolism in L6 myotubes, and of note, HIF-1α overexpression in macrophages enhanced IL-27 secretion, and increased MAFbx and MuRF-1 expression. IL-27 level was also elevated in the heart, serum, and gastrocnemius muscle of MI rats. Exercise counteracted these effects by promoting M2-like macrophage polarization and suppressing HIF-1α, thereby reducing IL-27 expression. Furthermore, exercise ameliorated IL-27-induced muscle atrophy via the WSX-1/gp130/pSTAT3 signaling axis. IL-27 contributes to muscle atrophy in post-MI cardiac insufficiency. Exercise attenuates IL-27-driven muscle wasting by modulating inflammation and promoting M2-like macrophage polarization. These findings provide insights into the mechanisms of MI-induced muscle atrophy and highlight the therapeutic potential of exercise in cardiac rehabilitation.

Ischemic heart disease, particularly myocardial infarction (MI), is a leading cause of global mortality, and the resulting myocardial fibrosis significantly impairs cardiac function, contributing to heart failure. Despite considerable efforts, effective therapeutic interventions for reversing myocardial fibrosis and restoring heart function remain elusive. Recent studies highlight the crucial role of metabolic reprogramming in the activation of cardiac fibroblasts (CFs) and their transformation into myofibroblasts, which drive fibrosis. Gastrodin (GAS), a phenolic glycoside derived from Gastrodia elata, has demonstrated promising anti-fibrotic and metabolic regulatory effects; however, its clinical application has been limited by poor pharmacokinetic properties and lack of targeted delivery. Here, we designed an intrapericardial (iPC) injection nanocomposite hydrogel incorporating FAP-targeted CAR-T cell membrane-coated GAS nanoparticles (FAP-CAR T CM@PPA-G NPs) within a pH/ROS-responsive CP/PCPD hydrogel. This innovative system achieves specific targeting of FAP-positive myofibroblasts, on-demand release of GAS triggered by pathological ROS and pH changes, and effective inhibition of fibrosis by modulating the KLF2/CREB5/HIF-1α/PFKFB3 metabolic axis. Our findings not only provide a novel therapeutic platform for post-MI fibrosis treatment but also elucidate the mechanistic basis of GAS's anti-fibrotic effects. This strategy offers significant promise for advancing the clinical translation of GAS-based therapies for myocardial fibrosis and heart failure.

Polystyrene micro/nanoplastics (PS-MPs/NPs) and the neonicotinoid imidacloprid (IMI) frequently co-occur in freshwater ecosystems, yet their combined toxicity profiles remain distinct and unresolved. Here, we exposed juvenile crucian carp (Carassius auratus) to PS-MPs (5 μm), PS-NPs (60-100 nm), and IMI, alone or in combination, to unravel their interactive mechanisms via a multi-omics approach. Results revealed a clear size-dependent toxicity pattern: while PS-NPs (especially with IMI) preferentially targeted the brain, PS-MPs (with or without IMI) induced the most severe intestinal histological injury, characterized by extensive intestinal fold atrophy and goblet cell depletion. In the brain, co-exposure to PS-NPs and IMI elicited potentiated neurotoxicity, manifesting as blood-brain barrier (BBB) breakdown, neuroinflammation, and a specific disruption of the glutamate-glutamine-γ-aminobutyric acid (Glu-Gln-GABA) metabolic cycle, which coincided with hyperactive and asocial behaviors. Microbiome analysis highlighted distinct dysbiotic signatures. Integrated network analyses further linked these gut microbial shifts to central neurochemical imbalances, implicating the microbiota-gut-brain axis as a potential pathway involved in systemic toxicity. In summary, this study differentiated between gastrointestinal damage caused by MPs and systemic combined toxicity caused by the penetration of NPs and IMI across biological barriers. It emphasizes the importance of size-specific assessment in understanding the complex risks of combined exposure to plastics and pesticides, providing insights for pollution management in agricultural hotspots.

Conventional needle-based vaccination may cause tissue injury and increase the risk of cross-contamination between animals. This study evaluated the immunological effects of delivering an inactivated porcine circovirus type 2 (PCV2) vaccine using a needle-free jet injection system compared with needle injection in pigs. Immune responses were assessed using enzyme-linked immunosorbent assay (ELISA) for PCV2-specific antibody levels on days 7, 14, and 28 post-vaccination. Cellular architecture at injection sites and differential gene expression were analyzed using immunohistochemistry and transcriptome sequencing. Results showed that pigs receiving needle-free vaccination at 0.50 mL and 0.75 mL injection volumes exhibited higher PCV2-specific antibody responses than pigs receiving 1.0 mL needle injection. Needle-free vaccination also preserved dermal collagen fiber architecture, altered antigen-presenting cell distribution, and was associated with distinct activation of immune-related pathways, particularly those involved in leukocyte migration and cytokine-cytokine receptor interactions. These findings indicate that needle-free vaccination was associated with distinct PCV2-specific antibody responses and local immune microenvironment changes, supporting its potential as an alternative vaccine delivery strategy for swine PCV2 vaccination.

In intensive aquaculture, post-larval Procambarus clarkii (the red swamp crayfish) are highly susceptible to stress — with oxidative stress as the primary form — induced by long-distance transport and high-density rearing, leading to substantial mortality and economic losses. Meanwhile, illegal use of diazepam to alleviate stress causes drug residues, threatening the safety of aquatic products. To address these industry challenges, this study selected four plant-derived compounds (isolated from plants) — gastrodin, matrine, paeonol, and eucalyptol — some of which have been proven to have sedative/stress-alleviating effects in other animals, and supplemented them into the diet of post-larval crayfish to explore their potential as natural alternatives to diazepam. A control group, four dietary supplement groups each at 20 mg/L, and a diazepam positive control group at 20 mg/L were established. After 21 days of rearing, which matches the pre-transport acclimation period for post-larvae, comprehensive evaluations were conducted through behavioral observation for sedative effect, antioxidant index determination, expression analysis of antioxidant-related genes (NRF2, HSP70, MT, FER), and intestinal section observation. The results showed that paeonol and eucalyptol exhibited both sedative effects and enhanced antioxidant capacity, and by regulating the activity of antioxidant enzymes such as catalase (CAT) and the expression of related genes, they improved the ability to alleviate stress; gastrodin only enhanced antioxidant capacity without significant sedative effects. No significant differences in growth performance were observed among all experimental groups and the control group. This study provides theoretical support for the healthy rearing of post-larval Procambarus clarkii, identifies that paeonol and eucalyptol have the potential to serve as alternatives to diazepam, and offers a feasible solution to reduce transport mortality and address diazepam abuse and drug residues.

We examined ruminal lamina propria stromal cells of 5-, 30-, and 60-day-old Boer goats using transmission electron microscopy and immunofluorescence to explore their functional morphology in pre-weaning lambs. At 5 days, cells were poorly differentiated, irregular, organelle-poor, and linked by short processes. By 60 days, fibroblasts, myofibroblasts, macrophages, and lymphocytes were observed, and junctional plaques and vesicles had emerged. Telocytes (TCs) with long, slender processes rich in rough endoplasmic reticulum and mitochondria were observed to release vesicles at 60 days. Immunofluorescence showed that processes of CD34+ vimentin+ cells lengthened with age, forming a dense network at 60 days. α-SMA+ cells were distributed continuously but did not co-localize with CD34+ cells. S100A4+ cells were scarce at 5 days and increased at 30 days. CD34+ S100A4+ cells accumulated at papilla bases, whereas CD34+ vimentin+ cells concentrated at both papilla tips and bases, implying a ruminal epithelial stem cell niche. The results reveal stromal remodeling during pre-weaning rumen development and provide morphological support for the role of stromal cells in regulating epithelial proliferation and differentiation.

Abstract Axially chiral biaryl dimethanols are ubiquitous and multifunctional intermediates for the synthesis of valuable atropisomeric molecules in advanced materials, drugs, natural products, and organocatalysts. Despite their broad synthetic utility, direct enantiodivergent catalytic access to this specific class of compounds remains largely unexplored, particularly in biocatalysis. Herein, we report a biocatalytic platform for enantiodivergent and atroposelective dynamic kinetic carbonyl reduction via transient seven-membered cyclic lactol intermediates. These engineered alcohol dehydrogenase (ADH)-driven transformations deliver up to 99% yield and enantioselectivity (>99:1 e.r. and < 1:99 e.r). This strategy demonstrates broad substrate compatibility, extending even to complementary “flipped” substrate series. Furthermore, the protocol is readily scalable to gram quantities and accommodates diverse downstream derivatizations. Mechanistic experiments and theoretical calculations delineate the pathway and illuminate the origin of enzymatic stereocontrol.

OBJECTIVES:To investigate the role of lactate in myocardial ischemia-reperfusion (MIR)-induced gastric mucosal injury. METHODS:Thirty C57BL/6J mice were randomized into sham-operated group, MIR (45 min ischemia and 2 h reperfusion) group, and MIR+LDHAI group, and gastric mucosal injury was assessed using macroscopic scoring and HE staining; serum lactate and cTnT levels were measured using ELISA. Metabolomics analysis was conducted to identify the differential serum metabolites between the sham-operated and MIR mice. In the cell experiment, human gastric mucosal cells (GES-1) were indirectly co-cultured with THP-1 cell-derived macrophages polarized to the M1 phenotype by lactate and/or lipopolysaccharide (LPS) stimulation. In both the gastric mucosa tissues of the mice and the co-cultured GES-1 cells, the protein expression levels of CD68, iNOS, Arg-1, COX-1, and COX-2 were detected using Western blotting and immunofluo-rescence staining. RESULTS:Compared to sham operation, MIR induced concurrent injury in both the heart and stomach of the mice and caused elevation of iNOS protein levels and reduction of Arg-1 protein expression in the gastric mucosa, where increased overlapping fluorescent areas were detected after MIR using fluorescent double-labeling for CD68 and iNOS. Serum metabolomics analysis revealed a significant increase in circulating lactate levels after MIR. In GES-1 cells, indirect co-culture with M1 macrophages induced obvious cell injury, decreased COX-1 expression, and increased cellular COX-2 protein expression. Western blotting and immunofluorescence staining confirmed that 15 mmol/L lactate significantly increased iNOS and decreased Arg-1 expression in THP-1-derived macrophages. Indirect co-culture of GES-1 cells with M1 macrophages induced by lactate or LPS resulted in obvious GES-1 cell injury. In the mouse models of MIR, inhibition of lactic acid production effectively alleviated MIR-induced gastric mucosal injury. CONCLUSIONS:Lactate-induced M1 macrophage polarization mediates MIR-related gastric mucosal injury in mice via the heart-gastric axis, an effect reversible by LDHA inhibition.

Telocytes (TCs), a distinct interstitial cell population, are increasingly being reported in the male reproductive system. This study aimed to confirm TC presence in goat testes, to characterize their immunophenotype and age-related differences. Testes from one-, two-, and twelve-month-old goats were examined by using transmission electron microscopy (TEM) and double immunofluorescence (DIF) staining. TEM revealed that TCs were widely distributed among peritubular myoid cells (PMC) in testes from 12-month-old goats and exhibited reduced cell bodies and cytoplasm compared to TCs in juvenile goats. Their telopodes (TPs) displayed the characteristic bead-like structure of alternating podomers (Pms) and podoms (Pds). The telopodes were slender and curved, intertwining near collagen bundles and blood vessels and releasing abundant vesicles. DIF staining reliably identified goat testicular TCs as CD34+/vimentin+ cells with elongated processes and spindle- or triangular-shaped bodies. TCs in 12-month-old goats were significantly longer than those in juvenile goats and had more elongated nuclei. They also exhibited markedly more protrusions and were arranged in four or five parallel stromal layers. The DIF findings corroborated the TEM observations. This study provides evidence of age-related morphological and distributional changes in goat testicular TCs, establishing a foundation for understanding their role in maintaining the microenvironmental homeostasis of the testes.

A recently identified mesenchymal cell population, telocytes (TCs), has been found in many tissues of different animal species. Jet needle-free injection (JNFI) is a promising non-invasive drug-delivery method that can trigger effective immune responses in the skin. In this preliminary morphological study, an inactivated porcine circovirus vaccine was delivered into pig neck skin by JNFI, and untreated normal neck skin served as the control. Histopathology, immunohistochemistry (IHC), immunofluorescence (IF), and transmission electron microscopy (TEM) were used to evaluate TC distribution, ultrastructure, and selected quantitative TEM parameters 24 h after injection. TCs were widely distributed in porcine skin, located between collagen fibers and around blood vessels, adipocytes, and sweat glands. They were also observed in contact with mast cells. TCs around sweat glands were CD34+, Vimentin+, and α-SMA+, whereas TCs at other sites were CD34+, Vimentin+, and α-SMA−. After JNFI, inflammatory cell infiltration into the skin was observed; TCs were in contact with these cells, and TCs surrounding adipocytes redistributed into the adjacent loose connective tissue. Quantitative TEM analysis showed that TC profile density and visible telopod length did not differ significantly between normal skin and JNFI 24 h skin (p ≥ 0.05). In contrast, vesicle profiles per TC increased significantly in both perivascular and adipose-associated compartments (p < 0.05). These findings provide morphological evidence suggesting that TCs may participate in early cutaneous responses after JNFI vaccine delivery.

T-2 toxin is a prevalent mycotoxin that poses a substantial threat to male reproductive health, while effective protective strategies remain limited. This study evaluated the protective effects of quercetin (Qu), a natural flavonoid, against T-2 toxin-induced testicular injury and investigated the underlying mechanisms, focusing on NLRP3 inflammasome-associated inflammation and fibrotic remodeling. Male ICR mice were exposed to T-2 toxin (1 mg/kg) for 28 days, with or without Qu (50 mg/kg) or the NLRP3 inhibitor MCC950 (20 mg/kg). Testicular morphology, TLR4/MyD88/NLRP3 pathway activation, pyroptosis-associated molecular changes, fibrotic remodeling, and blood-testis barrier (BTB)-associated proteins were assessed using histological staining, immunofluorescence, immunohistochemistry, and Western blotting. T-2 toxin exposure caused marked testicular histopathological alterations and activated the TLR4/MyD88/NLRP3 pathway, accompanied by increased IL-1β and IL-18 levels and alterations in pyroptosis-associated molecular markers in Leydig and Sertoli cells (P < 0.05). These changes were associated with increased TGF-β1 expression and peritubular collagen deposition, suggesting early fibrotic remodeling (P < 0.05). T-2 toxin also reduced the expression of the BTB-associated proteins ZO-1, occludin, CX43, claudin-1, and β-catenin (P < 0.05). Qu treatment attenuated testicular pathological changes and inflammasome-associated inflammatory responses, reduced early fibrotic remodeling, and restored BTB-associated protein expression. MCC950 intervention further supported the involvement of NLRP3 inflammasome-associated signaling in T-2 toxin-induced testicular injury. Collectively, these findings suggest that Qu alleviates T-2 toxin-induced testicular injury, at least in part, by modulating NLRP3 inflammasome-associated inflammation and fibrotic remodeling, thereby maintaining testicular homeostasis. Qu may therefore have potential for mitigating mycotoxin-induced reproductive toxicity. Nevertheless, because the conclusions regarding pyroptosis are based primarily on pathway-associated molecular markers in a murine model, further studies incorporating direct assessments of pyroptotic cell death and validation in higher-order animal models and clinical settings are warranted.

BACKGROUND:Netrin 4 (NTN4) has been reported to be involved in a variety of pathophysiological processes, such as the occurrence and development of tumors, viral replication and infection, and diabetic retinopathy. However, the relationships between NTN4 and metabolic diseases have not been reported. METHODS:The PhenoScanner tool and R language were used for bioinformatics analysis. Serum NTN4 was measured by ELISA in 211 healthy women and 193 overweight/obesity (OW/OB) women. Alterations in the serum NTN4 level were examined during the cold-exposure tests, acute exercise, lipid infusion, OGTT and EHC. RESULTS:GWAS and Bioinformatics analysis revealed that NTN4 was closely related to energy metabolism and OS. In a population-based cohort study, we observed that individuals with OW/OB and IR exhibited significantly elevated levels of circulating NTN4. In addition, the serum NTN4 concentration was found to be significantly correlated with indicators of obesity, IR, sex hormone and glucose/lipid metabolism. CONCLUSIONS:NTN4 is associated with obesity-related IR. TRIAL REGISTRATION:All research plans were approved by the Human Research Ethics Committee of Chongqing Medical University (No. (74)2012, No. (72)2014 and No. (74)2015).

Cardiac fibrosis (CF) is a common pathophysiological process in the development of various cardiovascular diseases, during which many cardiac fibroblasts undergo myofibroblast transdifferentiation. Fibroblast activation protein (FAP) can serve as a specific target for myofibroblasts, and chimeric antigen receptor (CAR)-based therapy is a promising immunotherapy strategy. In this study, we attempted to construct CAR natural killer (NK) cells that target FAP and explored their potential therapeutic role in CF. Our results suggested FAP CAR-NK-92 cells can specifically recognize and kill FAP+ cells in vitro. In addition, compared with parental NK-92 cells, FAP CAR-NK cells cocultured with FAP HEK-293 T cells presented increased cytotoxicity, cytokine secretion, and degranulation, indicating an effect-to-target ratio dependence. Coculturing FAP CAR-NK cells with mouse cardiac fibroblast lines (MCFs) eliminated the activated fibroblasts, reduced fibrosis-related protein secretion, and significantly reversed the contractile phenotype of myofibroblasts, which is characterized by alpha-smooth muscle actin (α-SMA) and stress fiber formation. Intravenous injection of FAP CAR-NK cells in mice 7 days after Ang II/PE-induced injury significantly improved cardiac function and reduced fibrosis. In terms of the killing mechanism, the early apoptosis rate of target cells was significantly increased, the antiapoptotic protein Bcl-2 was significantly decreased, and the proapoptotic proteins Bax and Caspase 3 were markedly increased. Our findings demonstrate that FAP CAR-NK-92 cells can specifically recognize FAP+ target cells and exert potent anti-fibrotic effects both in vitro and in vivo. Therefore, FAP CAR-NK-92 cells could be considered an effective therapeutic option for CF patients.

Teratoscincus roborowskii, an endemic species to China, is of particular interest due to its unique biological characteristics. It is a strictly nocturnal reptile that inhabits extremely harsh environmen. In this study, we generated the first chromosome level genome assembly of T. roborowskii using PacBio circular consensus sequencing (CCS) combined with Hi-C scaffolding. The genome assembly at the contig level was mounted onto chromosomes, with approximately 2,077.46 Mb of sequence allocated to 18 chromosomes, accounting for 99.92% of the total length. Among the sequences mapped to the chromosomes, approximately 2,070.96 Mb have determined order and orientation, making up 99.69% of the total length of the mapped sequences, with a contig N50 and scaffold N50 values of 117.38 Mb and 156.21 Mb respectively, and contains 21,341 predicted protein-coding genes, of which 99.62% were successfully annotated against public database. This high-quality genome not only fills a critical gap in genomic resources for this species, but also provides a valuable foundation for future study on its evolutionary history, gene functions and conservation biology.

Nail lichen planus (NLP) is a chronic inflammatory condition that can lead to considerable cosmetic and functional impairment. Failure to administer prompt and effective treatment may result in the development of permanent scarring and nail loss. The precise pathogenesis of NLP remains poorly understood, and there is currently an absence of safe and effective treatment options. Although not FDA-approved for the treatment of lichen planus, Janus kinase (JAK) inhibitors have shown considerable promise as therapeutic agents for a variety of dermatoses. This case report describes a patient with NLP who showed improvement after six months of treatment with upadacitinib, a selective JAK1 inhibitor. Changes were assessed using the Nail Lichen Planus Severity Index (NALSI) score. Following medication administration, the total score of the NALSI for the patient's nail involvement decreased from 146 to 37. However, a mild recurrence was observed following the reduction of the medication dosage (NALSI score to 47).

In therapeutic studies of pancreatic cancer, ultrasound-targeted microbubble destruction (UTMD) has shown potential in promoting apoptosis as a safe and non-invasive adjuvant therapy. Autophagy, a regulatory mechanism for cellular stress response and survival, plays a dual role in tumor development, progression, and treatment. However, the role of autophagy in UTMD-induced apoptosis in pancreatic cancer cells remains unclear. In this study, chloroquine (CQ), an autophagy inhibitor, was combined with UTMD to treat pancreatic cancer both in vitro and in vivo, with changes in apoptosis assessed through Western blot and TUNEL staining. The results showed that UTMD induced both apoptosis and autophagy in pancreatic cancer cells. Notably, inhibiting autophagy significantly enhanced UTMD-induced apoptosis, while the inhibition of apoptosis did not affect UTMD-induced autophagy. These findings suggest that autophagy reduces the effectiveness of UTMD in treating pancreatic cancer. This study offers a new perspective on UTMD for treating pancreatic cancer, suggesting that combining autophagy inhibitors could be a promising strategy to enhance the effectiveness of pancreatic cancer therapy.

Intervertebral disc degeneration (IDD), osteoarthritis (OA), and osteoporosis (OP) are common musculoskeletal disorders (MSDs) with similar age-related risk factors, representing the leading causes of disability. However, successful therapeutic development and translation have been hampered by the lack of clinically-relevant animal models. In this study, we investigated the potential suitability of the tree shrew, a small mammal with a close genetic relationship to primates, as a new animal model for MSDs. Age-related spontaneous IDD in parallel with a gradual disappearance of notochordal cells were commonly observed in tree shrews upon skeletal maturity with no sex differences, while age-related osteoporotic changes including bone loss in the metaphyses were primarily presented in aged females, similar to observations in humans. Moreover, in the osteochondral defect model, tree shrew cartilage exhibited behavior similar to that of humans, characterized by a more restricted self-healing capacity compared to the rapid spontaneous healing of joint surfaces observed in rats. The induced OA model in tree shrews was highly efficient and reproducible, characterized by gradual deterioration of articular cartilage, recapitulating the human OA phenotype to some degree. Surgery-induced IDD models were successfully established in tree shrews, in which the lumbar spine instability model developed slow progressive disc degeneration with more similarity to the clinical state, whereas the needle puncture model led to the rapid development of IDD with more severe symptoms. Taken together, our findings pave the way for the development of the tree shrew as a new animal model for the study of MSDs and aging.

BackgroundMyocardial ischemia-reperfusion injury (MIRI) frequently occurs during rapid restoration of blood flow in the infarcted myocardium. While Gastrodin (GAS) mitigates MIRI, its mechanism requires further exploration.MethodsWe evaluated GAS effect in SD rats following 45-min left coronary artery ligation and reperfusion. GAS (intraperitoneal) was administered preoperatively for 3 days. Triphenyltetrazolium chloride (TTC) staining was used to detect infarct size. The cardiac function was monitored by the Langendorff isolated cardiac perfusion system. Hematoxylin-Eosin (H&E) staining was applied to detect cardiac injury. H9c2 cells underwent oxygen and glucose deprivation (OGD) and were subsequently restored to normal culture conditions, mimicking MIRI. Cell Counting Kit-8 (CCK-8) was used to detect the cytotoxicity of GAS. Myocardial cell injury was determined by detecting lactate dehydrogenase (LDH) level in the medium. The expression of protein was detected by Western blot (WB) and immunofluorescence (IF) assay. Coimmunocoprecipitation (Co-IP), coupled with molecular docking detected the combination among transgelin2 (TG2), and CNPase.ResultsGAS reduced the size of myocardial infarction, alleviated myocardial fiber damage, and ameliorated MIRI-mediated cardiac dysfunction. Mechanistically, GAS inhibited apoptosis by restoring MIRI-altered TG2/CNPase expression. TG2 directly bound and negatively regulated CNPase. CNPase deficiency enhanced MIRI amelioration by reducing apoptosis.ConclusionTaken together, GAS protects against MIRI by modulating apoptosis through the TG2/CNPase pathway, revealing a novel therapeutic target.

Vascular calcification is tightly associated with cardiometabolic risk events. PDZK1 (PDZ domain containing 1) has been implicated in protecting from atherosclerosis, however, its relationship with vascular calcification remains unclear. Here we show that the expression levels of PDZK1 are notably boosted in calcified mice aortas, human and mouse vascular smooth muscle cells (VSMCs). PDZK1 deficiency in mice improves aortic calcification mediated by excessive vitamin D3 (VitD3). Consistently, blocking PDZK1 in human and mouse VSMCs effectively alleviates vascular calcification caused by high phosphate (Pi). PDZK1 overexpression in vitro promotes vascular calcification. Mechanistically, PDZK1 positively regulates the expression of β-catenin and its phosphorylated form at serine 552 (p-β-cateninSer552). Rescue experiments confirm that β-catenin is a critical mediator through which PDZK1 exacerbates vascular calcification. PDZK1 extends the half-life of β-catenin and p-β-cateninSer552 by blocking their ubiquitination and degradation. Furthermore, inhibition of PDZK1 attenuates high Pi-induced nuclear translocation of β-catenin whereas PDZK1 overexpression facilitates this process. More deeply, we reveal the direct interaction between PDZK1 and β-catenin is mediated by binding of β-catenin to the PDZ1 domain of PDZK1. These findings elucidate a novel PDZK1/β-catenin axis in vascular calcification progression and provide a potential therapeutic target for treating cardiovascular calcification in high-risk populations. Mechanistic insights into PDZK1-aggravated vascular calcification via β-catenin stabilization reveal a novel therapeutic target for prevention and treatment.

The aquaporin (AQP) gene family plays a critical role in water balance and osmotic regulation, yet its function and regulatory mechanisms in plateau reptiles remain poorly understood. In this study, we systematically identified 10 AQP genes in the Qinghai toad-headed agama (Phrynocephalus vlangalii) based on whole-genome data, and conducted a comprehensive analysis of their physicochemical properties, phylogenetic relationships, conserved domains, gene structures, and expression patterns. The results showed that the AQP genes of P. vlangalii are predicted to localize to the plasma membrane and exhibit significant tissue-specific expression, with the highest levels detected in the kidney and liver. Under low-temperature stress, multiple AQP genes displayed dynamic expression patterns during the stress and recovery phases. Specifically, AQP0, AQP2, and AQP5 were persistently downregulated in the liver, kidney, and brain, whereas AQP3, AQP7, and AQP9 were initially upregulated during early cold exposure but significantly downregulated during recovery, suggesting their coordinated roles in energy metabolism and water conservation. This study provides evidence supporting the involvement of the AQP gene family in the adaptation of P. vlangalii to the cold and arid plateau environment, providing new insights into the regulatory mechanisms of water metabolism in reptiles.