The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak has many unexpected implications, but the scientific community remains optimistic about overcoming these obstacles. Adenoviruses (Ad) are considered the most suitable vectors for transferring specific antigens to mammalian cells since they can induce both innate and adaptive immune responses. Ad-based coronavirus disease 2019 (COVID-19). vaccines were granted emergency use authorization in the COVID-19 pandemic. Many features of the Ad vector render it an appealing vaccine carrier for contagious diseases, including high titer, ease of processing, high effectiveness, low immunogenicity in clinical trials, and consistency in pharmaceutical packaging and shipment processes. Ad-based vaccines are generally effective and have few side effects since Ad induces minor infections in humans, and genetic modifications can block viral replication. These single-dose vaccines are effective not only in young individuals but also in adults. Clinical trials of these single-dose vaccines are commendable and have shown excellent safety and efficacy profiles. This review provides a summary of the development of single-dose vaccines against SARS-CoV-2.
Chronic kidney disease (CKD), driven by progressive renal fibrosis, lacks effective therapeutic targets. This study investigates thrombospondin-4 (THBS4) as a novel mediator of CKD-related fibrosis and explores its mechanistic basis.
This study collected 100 patients diagnosed with chronic kidney disease and 30 healthy individuals. Enzyme-linked immunosorbent assay (ELISA) analysis was conducted to assess the expression of THBS4 in CKD patients. Mouse unilateral ureteral obstruction (UUO) renal fibrosis model and Human Kidney-2 (HK2) cell fibrosis model were constructed to analyze the expression changes of THBS4 in renal fibrosis. To examine the effects of inhibiting THBS4 expression on the process of renal fibrosis, these two models were analyzed using Sirius red staining, Masson staining, immunohistochemistry, real-time quantitative PCR (qPCR) and western blot methods.
The expression of THBS4 in the serum of CKD patients was found to be significantly higher (p < 0.05), and its concentration showed a negative correlation with the eGFR levels (r = –0.77, p < 0.05) and an increase corresponding to the progression of CKD stages (p < 0.05). THBS4 expression was dramatically increased in UUO-treated mouse kidneys as well as in TGF-β1-stimulated HK2 cells (p < 0.05). In vitro, the expression of renal fibrosis-associated proteins was also significantly reduced after interfering with THBS4 expression (p < 0.05). UUO-induced renal fibrosis and related protein expression were suppressed in THBS4 knockdown mice when compared to control mice (p < 0.05). The levels of p-AKT and p-PI3K exhibited a significant rise in conjunction with the onset of renal fibrosis (p < 0.05). The expression of p-AKT as well as p-PI3K showed a significant reduction upon inhibition of THBS4 expression (p < 0.05). Insulin-like growth factor 1 (IGF-1) treatment reversed these effects.
THBS4 was significantly overexpressed in CKD patients. By suppressing the expression of proteins associated with renal fibrosis and inhibiting the activation of the PI3K/AKT pathway, THBS4 has the potential to mitigate renal fibrosis.
Alpha-synuclein (α-syn) has long been identified as the etiologic agent of multiple neurodegenerative diseases, the most common and well-known of which are Parkinson’s disease (PD) and Lewy body dementia (LBD). While it is known that the pathophysiology of these synucleinopathies involves aggregation of improperly-folded α-syn, the mechanisms leading to its accumulation have not been fully identified. However, multiple pathways have been proposed, any or all of which may contribute to synucleinopathies. The role of α-syn in normal homeostasis and in other organ systems, especially the hematopoietic system, has been reported recently. Research within the last decade has shown that α-syn plays many vital and conserved roles in the cell biology of various organ systems, such as packaging of cell products, exocytosis, membrane stabilization, and more. This protein has been recognized as an essential factor in normal hematopoietic and immune systems function, and its deficiency leads to an abnormal phenotype, in hematopoietic and immune cell lineages. Similar phenotypes in synucleinopathies not only emphasize the conserved nature of the synuclein family but suggest a bimodal pathophysiology in which aggregated α-syn leads to cellular toxicity while causing derangement of systems that require it. Research into specific molecular mechanisms and potential treatments may provide further understanding of neurodegenerative diseases as well as lead to novel therapies. However, elucidation of the systemic roles of α-syn in addition to its toxicity in excess is essential to prevent treatment-induced deprivation, which paradoxically harms the patient. Here, we address recent advances in systemic synucleinopathies and putative interconnectedness of these compartments. While previous studies and reviews have focused on the mechanisms of α-syn synthesis, transport, and aggregation within systems, this review focuses on the potential inter-systemic nature of synucleinopathies and their possible synergistic origins.
This review explores the structure of polyamines, including putrescine, spermidine, and spermine, and their crucial roles in immune cell functions. Polyamines are active compounds derived from ornithine that regulate signaling pathways by interacting with nucleic acids and proteins. Polyamines are essential for normal growth and development in immune cells, participating in cell signaling and neurotransmitter regulation and playing a critical role in immune responses. Notably, high concentrations of polyamines play a significant role in tumor cells and autoreactive B and T cells in autoimmune diseases. This impact should not be overlooked. Elevated levels of polyamines are associated with enhanced immune cell activity in tumor cells and autoimmune diseases. Furthermore, the connection between polyamines and normal immune cell functions, as well as their roles in autoimmune and antitumor immune cell functions, is significant. The role of polyamines in the normal function of activated T cells is well-established, and they are particularly important in antitumor immunity by modulating immune cell functions in the tumor microenvironment (TME). By synthesizing the latest research advancements, this review provides valuable insights into the roles of polyamines in immune regulation and outlines directions for future research.
Cardiovascular diseases (CVD) remain the leading cause of global mortality, highlighting the urgent need for the identification of novel biomarkers and the development of therapeutic approaches to improve patient outcomes. Despite great progress in CVD diagnosis, treatment, and predicting risk, current methods fall short of effectively reducing its prevalence. Recently, bone morphogenetic protein 10 (BMP10), a cardiac-specific growth factor with a role in cardiac development and vascular homeostasis, has emerged as a potential biomarker and therapeutic target in CVD. While studies have demonstrated BMP10’s diagnostic potential in atrial fibrillation (AF), its precise role across the broader CVD landscape remains poorly understood. We review the current knowledge of BMP10’s involvement across a spectrum of cardiovascular conditions, including AF, heart failure, myocardial infarction, pulmonary arterial hypertension, dilated cardiomyopathy, and diabetic cardiomyopathy. This analysis provides an in-depth examination of the mechanisms through which BMP10 may influence CVD progression and highlights its potential utility as a diagnostic and therapeutic target.
Lung cancer remains a leading cause of cancer-related mortality due to its capacity for silent metastasis and the significant challenges in achieving effective treatment. Currently, targeted therapies and chemotherapies are the primary options for advanced or inoperable lung cancer; however, their efficacy is often undermined by the cancer’s ability to develop resistance through both genetic and non-genetic mechanisms. This review explores recent advances in understanding metabolic reprogramming in non-small cell lung cancer (NSCLC), focusing on its critical role in cancer progression. NSCLC cells exhibit heterogeneous activation of metabolic pathways influenced by their oncogenic mutations. Notably, their metabolic phenotypes evolve in response to environmental stressors and therapeutic pressures. Moreover, NSCLC cells engage in metabolic crosstalk with their microenvironment to enhance survival, leveraging distinct metabolic adaptations at both primary and metastatic sites. Despite extensive preclinical studies evaluating novel therapeutic strategies targeting these metabolic pathways, many have failed in clinical trials due to severe adverse effects. This is because the targeted pathways are crucial not only for cancer cells but also for normal cellular functions. Future research must prioritize approaches that selectively disrupt cancer-specific metabolic regulation to improve therapeutic outcomes.
Parasitic diseases, caused by a diverse array of parasites, remain a substantial threat to global health. Toll-like receptor 3 (TLR3) represents a pivotal element in the innate immune system, distinguished by an ability to signal via the TIR-domain-containing adapter-inducing interferon-β (TRIF)-dependent pathway upon detecting pathogen-derived double-stranded RNA (dsRNA), exosomal RNA (exoRNA), and long non-coding RNA (lncRNA). Predominantly localized on endosomal membranes, TLR3 is extensively expressed in neurons, immune cells, fibroblasts, and epithelial cells. Upon activation, TLR3 engages adaptor molecules such as TRIF, facilitating the phosphorylation of TANK-binding kinase 1 and the subsequent activation of interferon regulatory factors. This signaling cascade triggers the production of type I interferons (IFN-α/β) and proinflammatory cytokines such as interleukin (IL)-6, IL-8, IL-12, and tumor necrosis factor-alpha, which are crucial for effective immune defense against infections. Recent findings highlight the essential role of TLR3 in parasitic infections by detecting nucleic acids from damaged cells to activate dendritic and natural killer cells. TLR3 also functions with other receptors, such as TLR2 and TLR4, to enhance cytokine production and improve parasite clearance. However, TLR3 overactivation can induce excessive, harmful inflammation and tissue damage, highlighting its dual role in balancing immune defense. This review comprehensively examines the TLR3 signaling pathway and its multifaceted role in various parasitic infections, including those caused by Plasmodium spp., Leishmania spp., Clonorchis sinensis, Schistosoma japonicum, Trichinella spiralis, and Neospora caninum.
Memory T cells are essential for effective and durable immune responses, as they provide long-term immunological surveillance and rapid reactivity upon re-exposure to a given pathogen or cancer cell. In solid tumors, the immunosuppressive tumor microenvironment (TME) often hinders immune activation, making enhancing memory T cell formation and persistence a key goal in cancer immunotherapy. Novel strategies are exploring ways to support these memory T cells, including using Listeria monocytogenes as a cancer vaccine vector. Notably, L. monocytogenes has unique properties that make it an ideal candidate for this purpose: it is highly effective at activating T cells, promoting the differentiation and survival of memory T cells, and modulating the TME to favor immune cell function. Thus, by leveraging the ability of L. monocytogenes to induce a strong, sustained T-cell response, researchers aim to develop vaccines that provide lasting immunity against tumors, reduce recurrence rates, and improve patient survival outcomes. This mini-review highlights the potential of memory T cell-focused cancer immunotherapy and the promising role of L. monocytogenes in advancing these efforts.
Adipose-derived stem cells (ASCs) have been extensively investigated for regeneration and tissue engineering applications owing to their inherent regenerative ability. However, the effects of various species and depot-specific extraction sites on ASC differentiation and renewal capacity have yet to be explored thoroughly, limiting the clinical use of ASCs. Despite promising clinical results, ASCs are also associated with poor disease outcomes, specifically in the context of breast cancer and obesity. Only when ASC-driven obesity and breast cancer are understood separately will the connection between the two diseases and the ASC-associated effects therein be fully established. Therefore, this review aimed to assess the behavioral differences of ASCs from various large mammalian species and human-derived anatomical niches. This review analyzes ASC migration, the role of ASCs in breast cancer progression and immune modulation, and breast cancer-driven ASC dysfunction to further the understanding of ASCs for future clinical applications.
Adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) is an energy homeostasis controller that regulates various metabolic pathways to promote adenosine triphosphate (ATP) generation and suppress energy expenditure, thereby restoring energy homeostasis. As a co-factor in many enzymes, iron is an essential mineral for maintaining ATP levels in our bodies. Ferroptosis is an iron-dependent mode of cell death that occurs in various pathological processes, including cancer, metabolic disorders, and autoimmune diseases, by regulating iron metabolism, lipoperoxidation, and anti-oxidation functions. Ferroptosis is triggered by oxidative and energy stress, both controlled by cancer-associated signaling pathways. Emerging studies have demonstrated that AMPK directly influences ferroptosis by modulating lipid metabolism, redox homeostasis, and iron transport. Cancer cells exhibiting elevated baseline AMPK activity demonstrate resistance to ferroptosis, whereas AMPK suppression enhances their susceptibility to this regulated form of cell death. While the precise mechanistic details are yet to be fully elucidated, accumulating evidence suggests that AMPK-mediated ferroptosis regulation may contribute to cancer development and therapeutic responses. This review summarizes recent advances in understanding the interplay between AMPK and ferroptosis in cancer biology and discusses the potential of targeting the AMPK-ferroptosis axis for innovative anticancer strategies.
Pulmonary fibrosis is a life-threatening progressive lung disease characterized by increased fibrogenesis and decreased lung function. Pulmonary fibrosis has a poor prognosis and a low patient survival rate, with no effective treatments currently available. Cellular senescence is thought to contribute to the pathogenesis of this aging-related disease. Cellular senescence and premature aging are involved in the development of pulmonary fibrosis, which affects various cellular processes such as proliferation, apoptosis, and inflammatory responses. Multiple pathways contribute to cellular senescence and participate in the pathogenesis of pulmonary fibrosis, such as tumor protein p53 (p53)/cyclin dependent kinase inhibitor 1A (p21) and cyclin dependent kinase inhibitor 2A (p16)/retinoblastoma protein (pRB). However, many unanswered questions remain concerning the relationship between cellular senescence and pulmonary fibrosis. In this review, we first summarize the common causes of lung cell senescence and pulmonary fibrosis, including aging, inflammation, chemotherapy drugs, and environmental pollutants. We also discuss the enriched signaling pathways and epigenetic factors (e.g., non-coding RNAs) that are important during cell senescence and the progression of pulmonary fibrosis. Finally, we discuss current strategies for treating pulmonary fibrosis by targeting cellular senescence, including relevant preclinical and clinical studies. This review provides new mechanistic insights for understanding the role of cellular senescence in the development of pulmonary fibrosis and its treatment by targeting cellular senescence.
Bronchoalveolar lavage (BAL) constitutes a valuable diagnostic approach for the differential diagnosis of various pulmonary fibrotic diseases. BAL fluids from patients with interstitial lung diseases (ILDs) can also be utilized for research purposes, offering cell populations suitable for functional and phenotypical studies. In this study, we demonstrate the feasibility of isolating a discrete number of fibroblasts/myofibroblasts in vitro from the BAL fluid from ILD patients, a procedure typically performed during the early stages of disease when high-resolution computed tomography does not yield a definitive diagnosis.
We obtained BAL samples from a total of 43 patients. Fibroblasts were successfully derived in vitro from 20 patients, with larger quantities of cells from 11 patients. Whenever possible, the cells were cultured and expanded until passage 12–15. Fibroblasts could be expanded to passage 36 in only one case. The expression of typical fibrotic markers, such as type I collagen, α-smooth muscle actin, and fibronectin-extra domain A or B (FN-EDA/-EDB), was therefore compared in fibroblasts obtained from ILD-patients with fibroblasts derived from non-diseased controls by quantitative RT-PCR, immunofluorescence, and cytofluorographic analysis. The rate of proliferation, migration, and response to the anti-fibrotic drug pirfenidone was further determined in 2D and in 3D models of in vitro cultures.
A specific morphological heterogeneity among fibroblasts/myofibroblasts derived from patients with fibrotic or non-fibrotic ILD was observed, such as enlarged and flattened shaped cells vs spindle-shaped cells. Moreover, a higher expression of α-smooth muscle actin (α-SMA), type I collagen (collagen I), and fibronectin was demonstrated in ILD fibroblasts than in control fibroblasts. The anti-fibrotic drug pirfenidone was effective in inhibiting the growth and migration of ILD-fibroblasts both in 2D and 3D in vitro models.
Collectively, the present study suggests that BAL-derived fibroblasts from ILD patients may serve as a useful in vitro model for studying and assaying pulmonary fibrosis. This approach has the potential to improve our understanding of ILD pathogenesis and overcome ethical and availability concerns associated with biopsy-derived tissues.
In discussing the interplay between light exposure/blue light exposure (LE/BLE) and circadian health, we emphasize the role of light hygiene and its effects on maintaining sleep, mood, and metabolic health, among other physiological processes. We define compromised circadian light hygiene as low dynamic range and/or irregular 24-hour patterns of LE. Poor light hygiene interferes with circadian entrainment and weakens circadian robustness alignment, thereby increasing health hazards. We provide an overview of the complex molecular pathways underlying light perception and downstream signaling. Given that genetic polymorphisms influence key elements within these light signaling pathways, we propose that personalized light hygiene approaches be designed for populations affected by compromised circadian LE or at risk of light-induced circadian disruption.
Infection with Staphylococcus aureus (S. aureus) is an important contributor to intervertebral disc degeneration (IDD). Endoplasmic reticulum stress (ERS) is a major pathway through which bacteria regulate cell fate. The aim of this study was to examine the role of ERS in S. aureus-induced IDD.
We assessed the S. aureus-induced degeneration, apoptosis, and senescence of nucleus pulposus cells (NPCs) in vitro by Western blot, flow cytometry, and staining for β-galactosidase, and in vivo by magnetic resonance imaging/computed tomography (MRI/CT) imaging, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and histological staining. RNA sequencing was conducted to identify differentially expressed genes, while siRNA, lentiviral vectors, and Atf3-knockout (Atf3-KO) mice were utilized to confirm the role of ATF3 in persistent IDD following transient S. aureus infection.
Following the eradication of S. aureus in vitro, the expression of Aggrecan and collagen II in NPCs continued to decline, accompanied by an increase in the proportion of apoptotic and senescent cells. Transient S. aureus infection was shown to activate the Activating Transcription Factor 3 (ATF3)-CCAAT/Enhancer-Binding Protein Homologous Protein (CHOP) signaling pathway, leading to sustained swelling of the endoplasmic reticulum in NPCs. In vivo experiments further demonstrated that transient S. aureus infection resulted in progressive IDD, activation of the ATF3-CHOP pathway, increased numbers of TUNEL-positive cells, and elevated P21 expression. Knockdown of ATF3 expression in vitro attenuated the S. aureus-mediated increase in apoptotic and senescent cells, while Atf3-KO mice exhibited milder IDD compared to wild type (WT) mice, with fewer apoptotic cells and reduced P21 expression.
Transient S. aureus infection may lead to progressive IDD by triggering sustained ER stress and activating related signaling pathways. The ATF3-CHOP pathway may be an important target for alleviating the sustained disc degeneration caused by transient S. aureus infection.
The metabolites derived from judicious dietary choices play a crucial role in the management and treatment of depression. Hydroxy-carboxylic acid receptor 2 (HCAR2) functions as a receptor for various diet-derived metabolites. Although a growing body of evidence indicates these metabolites exert beneficial effects on depression, the precise mechanisms underlying these benefits require further investigation.
We established a mouse model of corticosterone (Cor)-induced depression to evaluate the therapeutic potential of HCAR2 activation on depression. A series of behavioral experiments were conducted to investigate whether HCAR2 activation could alleviate depressive-like behaviors in mice. The neuroprotective effects of HCAR2 in the hippocampus were examined using Nissl and hematoxylin-eosin (HE) staining. The levels of monoamine neurotransmitters in mouse serum were quantified, as well as the cell viability and lactate dehydrogenase (LDH) activity of hippocampal neurons co-cultured with primary microglia. Microglia-associated neuroinflammation was evaluated by quantifying pro-inflammatory cytokines using ELISA, and by assessing the polarization state of M1 microglia, including the mRNA expression levels of M1 markers and double fluorescence staining for inducible nitric oxide synthase/ionized calcium-binding adapter molecule 1 (iNOS/Iba1). The expression level of proteins in the protein kinase B-inhibitor of nuclear factor kappa-B kinase subunits alpha and beta-nuclear factor kappa-light-chain-enhancer of activated B cells (AKT-IKKαβ-NFκB) pathway in primary microglia was analyzed using western blot. Transcriptomic changes in microglia induced by HCAR2 activation were examined through RNA sequencing. Mice were fed PLX5622 chow to deplete microglia in vivo.
Activation of HCAR2 by its agonist MK-6892 in a Cor-induced model of depression significantly alleviated depressive-like behaviors, attenuated hippocampal neuronal injury, increased serum monoamine levels, reduced microglia-associated neuroinflammation, and inhibited the expression of proteins in the AKT-IKKαβ-NFκB pathway in primary microglia. Additionally, HCAR2 activation markedly enhanced hippocampal neuronal viability and decreased LDH activity in this co-culture system. Importantly, these protective effects were abolished in HCAR2 knockout mice. RNA sequencing revealed that HCAR2 activation induced changes in multiple signaling pathways. Moreover, the depletion of microglia also eliminated the protective effects of MK-6892.
Activation of HCAR2 can reduce depressive-like behaviors, neuronal injury, and neuroinflammation. Our findings suggest these neuroprotective effects are, at least in part, mediated through modulation of microglial activity by HCAR2.
Epithelial ovarian cancer (EOC) is a highly lethal gynecological malignancy characterized by frequent late-stage diagnosis, high rates of chemoresistance, and poor long-term survival. Emerging evidence underscores the central role of iron metabolism dysregulation in EOC pathogenesis, progression, and treatment resistance. Ovarian cancer cells and cancer stem cells exhibit an “iron-addicted” phenotype, characterized by increased iron uptake, reduced export, and enhanced storage, which sustains proliferative signaling, redox imbalance, and metastatic potential. Recent advances have illuminated ferroptosis, a regulated form of iron-dependent cell death driven by lipid peroxidation, as a promising therapeutic target for overcoming resistance to platinum-based chemotherapy. This review provides a comprehensive synthesis of the mechanisms governing iron metabolism and ferroptosis in EOC, with a particular focus on Class IV ferroptosis inducers (FINs). These agents act by disrupting iron homeostasis and promoting labile iron pool accumulation, thereby triggering oxidative stress and ferroptotic death. Preclinical studies demonstrate that Class IV FINs, including iron nitroprusside, superparamagnetic iron oxide nanoparticles, ferric ammonium citrate, and Ferlixit, exhibit potent antitumor activity in EOC models, particularly in chemoresistant and stem-like tumor subpopulations. Furthermore, Class IV FINs show synergistic effects when combined with other ferroptosis modulators or immunotherapeutic agents. Despite their promise, clinical translation remains limited by challenges in bioavailability, delivery specificity, and potential systemic toxicity. Ongoing efforts in nanotechnology, biomarker discovery, and tumor stratification offer new avenues for refining ferroptosis-based interventions. Ultimately, this review highlights Class IV FINs as a mechanistically distinct and clinically actionable strategy to target metabolic vulnerabilities in EOC, with the potential to reshape therapeutic paradigms and improve patient outcomes.
Heart regeneration requires renewal of lost cardiomyocytes. However, the mammalian heart loses its proliferative capacity soon after birth, and the molecular signaling underlying the loss of cardiac proliferation postnatally is not fully understood.
This study aimed to investigate the role of Catenin alpha 3 (Ctnna3), coding for alpha T catenin (αT-catenin) protein in regulating cardiomyocyte proliferation and heart regeneration during the neonatal period.
Here we report that ablation of Ctnna3 and highly expressed in hearts, accelerated heart regeneration following heart apex resection in neonatal mice.
Our results show that Ctnna3 deficiency enhances cardiomyocyte proliferation in hearts from postnatal day 7 (P7) mice by upregulating Yes-associated protein (Yap) expression.
Our study demonstrates that Ctnna3 deficiency is sufficient to promote heart regeneration and cardiomyocyte proliferation in neonatal mice and indicates that functional interference of α-catenins might help to stimulate myocardial regeneration after injury.
Nitrate transporter NRT1/PTR family (NPF) proteins are crucial for plant nitrogen uptake and utilization. As an important hexaploid crop for grain and forage, oat (Avena sativa L.) requires substantial levels of nitrogen. However, the oat nitrate transporter 1 (NRT1) family remains uncharacterized.
In this study, the oat NRT1 subfamily members were identified through the Hmm and Pfam databases. Bioinformatics analysis was performed using the MEGA 11 and TBtools software to elucidate the physicochemical properties, evolutionary relationships, chromosomal localization, and gene structures. Furthermore, quantitative real-time polymerase chain reaction (qRT-PCR) analysis and the green fluorescent protein (GFP) fusion expression vector were utilized to investigate the candidate oat NRT1s.
Phylogenetic classification categorized oat NRT1s into eight subfamilies, with the most abundant being the NPF5 subfamily. Physicochemical property analysis revealed that the number of amino acids in the proteins encoded by these genes ranged from 235 to 673, with their molecular weights (MWs) ranging from 26 kDa to 74 kDa. Chromosomal localization revealed that these genes were unevenly distributed across all 12 oat chromosomes. Promoter analysis revealed that light-responsive elements appeared most frequently in the promoters of these genes (39.3%), followed by abscisic acid (ABA)-responsive elements (13.5%) and methyl jasmonate (MeJA)-responsive elements (9.4%). qRT-PCR analysis revealed that most of the genes exhibited tissue-specific expression patterns. Among them, AsNPF2.6 was highly expressed in the leaves at 1 h post-low nitrogen (LN) treatment, while AsNPF4.5 was highly expressed in the leaves at 12 h. Both these genes exhibited low expression levels in the roots. However, AsNPF7.16 and AsNPF7.19 were both highly expressed in the roots at 9 h post-LN treatment but exhibited low expression in the leaves. Subcellular localization revealed that all five proteins (AsNPF2.6, AsNPF4.5, AsNPF7.16, AsNPF6.8, and AsNPF7.19) were localized to the cytoplasm and cell membrane.
Our results demostrate the involvement of AsNRT1 family members in nitrogen transport in oat, providing theoretical support for further investigation into the functions and molecular mechanisms of action of oat NRT1s in nitrogen transport.
Ion and water transport in the central nervous system (CNS) is governed by tightly coupled mechanisms involving electrodiffusion, osmotic pressure, and fluid convection. Disruptions to these processes are implicated in pathological conditions. Understanding the coordinated roles of glial cells and perivascular spaces in regulating ionic and fluid homeostasis is essential for interpreting neural function and dysfunction.
We developed a multicompartment model of the optic nerve incorporating axons, glial cells, extracellular space (ECS), and three perivascular compartments (arterial, venous, and capillary-associated). The model integrates electrodiffusion of ions, osmotic water transport, and convection, while enforcing electroneutrality and compartmental volume conservation. Numerical simulations were performed using a finite volume method under axisymmetric geometry, and parameter sensitivity was explored through variations in glial membrane conductance, connexin permeability, and aquaporin-4 (AQP4) expression.
The simulations reveal that potassium released from axons during stimulation is cleared via glial uptake and redistributed through electric drift within glial syncytia. The perivascular pathway provides a secondary route for potassium and water clearance. Decreased glial conductance leads to abnormal firing in unstimulated axons, mimicking epileptiform activity, while reduced connexin coupling increases dependence on perivascular drainage. Changes in AQP4 expression had limited effect on ionic homeostasis in the current model.
This model provides a biophysically consistent framework to study ionic-fluid coupling in CNS microcirculation. It demonstrates how glial and perivascular compartments cooperate to maintain extracellular potassium balance. The findings offer insight into the mechanisms underlying pathological K+ accumulation and suggest potential therapeutic targets involving glial modulation and perivascular enhancement. The framework is extensible to other brain regions and conditions involving impaired clearance or excitability.
Hypokalemia induces abnormal spontaneous pacemaker activities of cardiomyocytes, which is strongly associated with fatal cardiac arrhythmias caused by hypokalemia. However, the mechanism remains unclear.
For the study of the mechanisms associated with hypokalemia, optical mapping recordings were performed on isolated murine hearts perfused with hypokalemia solutions, which allows for the concurrent examination of membrane potential and calcium transient morphology and arrhythmogenesis. Human Kir2.1, Kir2.1-E224G mutant, or Kir4.1 channels were constructed with lentiviral vectors. Patch clamp recordings were performed to verify the corresponding currents of these constructed channels in the heterologous expression system chinese hamster ovary (CHO) cells, and to explore how Kir2.1 channels influence the resting membrane potentials of human iPSC-derived cardiomyocytes (hiPSC-CMs) when exposed to low [K+]e.
Isolated murine hearts perfused with hypokalemia solution (1 mmol/L) developed a high frequency of spontaneous ventricular tachycardia (VT), which was initiated as an after-depolarization triggered activity associated with Ca2+ overload. The VT was maintained by abnormal spontaneous pacemaker activities caused by membrane potential depolarization. In response to 1 mmol/L [K+]e, hiPSC-CMs overexpressing Kir2.1 channels exhibited membrane potential depolarization, leading to the induction of abnormal pacemaker activities. The cells overexpressing rectification-deficient Kir2.1-E224G mutant channels or weak rectification Kir4.1 channels exhibited membrane potential hyperpolarization without the occurrence of abnormal pacemaker activities.
Kir2.1 channel-mediated membrane potential depolarization contributes to hypokalemia-induced abnormal spontaneous pacemaker activities of cardiomyocytes. The inward rectification of Kir2.1 channels plays a critical role in this process.
Quercetin, a naturally occurring flavonoid, possesses anti-inflammatory properties and has emerged as a potential modulator of tissue repair. Impaired wound healing and pathological scarring are often driven by excessive inflammation and dysregulated myofibroblast differentiation. Current therapeutic approaches, however, frequently fall short in simultaneously addressing these intertwined challenges. This study investigates whether quercetin can provide a bifunctional therapeutic advantage by promoting early wound closure through inflammation resolution and suppressing scar formation via the inhibition of myofibroblast differentiation.
A murine excisional wound model was employed to evaluate quercetin’s effects in vivo. Mice (C57BL/6, n = 8/group) received daily topical applications of 1% quercetin. Wound closure kinetics were meticulously quantified using planimetry. To assess molecular and cellular changes, protein levels (CASPASE-1, interleukin-1 beta (IL-1β), alpha-smooth muscle actin (α-SMA)) and collagen III/I ratios were determined through multiplex qPCR, RNA sequencing, western blot analysis, and histomorphometry. For in vitro investigations, human dermal BJ fibroblasts were treated with transforming growth factor beta 1 (TGF-β1) (10 ng/mL) ± quercetin (5–50 μM) to assess myofibroblast differentiation markers (α-SMA, collagen I) via immunofluorescence, western blot, and qPCR.
Quercetin significantly accelerated wound closure in vivo. The acceleration was accompanied by a reduction in the expression of IL-1β and CASPASE-1. RNA sequencing data revealed that quercetin’s anti-inflammatory effects in early wound healing involve the modulation of inflammasome complexes, including NLRP3, as well as inflammasome-mediated signaling pathways. Furthermore, treated wounds exhibited increased collagen III/I ratios relative to control groups (p < 0.05), indicative of a more regenerative matrix remodeling process. In vitro, experiments demonstrated that quercetin suppressed TGF-β1-induced myofibroblast differentiation, evidenced by decreased α-SMA expression (p < 0.05) and reduced collagen I synthesis. Notably, quercetin exhibited cell type-specific effects: while suppressing BJ fibroblast migration (scratch assay), it enhanced keratinocyte proliferation. This unique duality prevents aberrant myofibroblast recruitment without compromising essential epithelial coverage—a critical balance for minimizing scar formation.
Quercetin exhibits a compelling dual therapeutic role in wound healing: resolving inflammation to expedite early wound healing and inhibiting TGF-β-driven myofibroblast differentiation to attenuate scarring. By harmonizing these actions, quercetin addresses both phases of repair, positioning it as a promising candidate for scar-free wound therapy. Further efforts should focus on optimizing its bioavailability to enhance clinical translation.
In mammals, skeletal muscle typically constitutes approximately 55% of body weight. The thermogenesis of skeletal muscle increases with increased cold stress, and skeletal muscle maintains the animal’s body temperature through the heat generated by shivering. However, less attention has been paid to investigating the impact of cold stress on the fiber type makeup of skeletal muscle, especially the gastrocnemius. Consequently, this research explored how cold stress regulates muscle development and fiber type composition.
A cold stress model was established by subjecting mice to a 4 °C environment for 4 hours daily. This model was combined with an in vitro siRNA-mediated knockdown model for joint validation. The impact of cold stress on skeletal muscle development and myofiber type transformation was assessed using experimental techniques, including immunofluorescence and western blotting.
Following cold stress, the expression level of Myosin Heavy Chain 7 (MYH7) in the mouse gastrocnemius increased, while Myosin Heavy Chain 4 (MYH4) expression decreased. Concurrently, elevated expressions of Mindbomb-1 (Mib1) and the myogenic differentiation (MyoD) were observed. Subsequent knockdown of Mib1 in C2C12 cells resulted in increased MYH4 expression and decreased MYH7 expression.
Cold stress induces skeletal muscle fibers to shift from fast-twitch to slow-twitch through the Mib1/Notch signaling pathway.
Inflammation plays a pivotal role in the progression of tissue fibrosis. Our previous research demonstrated that Na+, K+-ATPase (NKA) α1 deficiency impairs mitochondrial function and accelerates isoproterenol (ISO)-induced cardiac remodeling. This study aims to investigate the interplay between inflammation and NKAα1 deficiency in ISO-induced cardiac fibrosis.
Age-matched male wild-type (WT) and NKAα1+/- mice received daily subcutaneous injections of ISO (30 mg/kg body weight) over 14 consecutive days. Comprehensive histopathological evaluation was performed to assess myocardial architecture and leukocyte infiltration profiles. Mitochondrial ultrastructure was analyzed using transmission electron microscopy. The molecular techniques of real-time quantitative polymerase chain reaction (RT-qPCR), immunoblotting, and enzyme-linked immunosorbent assay (ELISA) were utilized to quantify fibrotic markers and inflammatory mediators. A cell co-culture model was established to investigate the interactions between different cell types.
NKAα1 haploinsufficiency exacerbated heart lesions and fibrosis, led to macrophage accumulation, and increased the expression of inflammatory factors in ISO-challenged hearts. Although NKAα1 deficiency did not directly activate macrophages or fibroblasts under ISO conditions, it significantly accelerated cardiomyocyte death in response to ISO insult. Paracrine crosstalk between damaged NKAα1+/- cardiomyocytes, macrophages, and fibroblasts amplified macrophage activation, inflammatory cytokine release, and fibroblast differentiation. Estrogen-related receptor α (ERRα) was identified as a key mediator of NKAα1 haploinsufficiency-induced cardiomyocyte death and interleukin-18 (IL-18) release. Furthermore, treatment with an NKAα1 897DVEDSYGQQWTYEQR911 (DR)-region antibody mitigated ISO-induced cardiac fibrosis and macrophage infiltration.
This study provides evidence that NKAα1 deficiency exacerbates cardiac fibrosis by promoting ERRα-dependent cardiomyocyte death and by facilitating intercellular cross-talk between damaged NKAα1+/- cardiomyocytes, macrophages, and fibroblasts. Based on these findings, we suggest that NKAα1 may be a potential regulator of cardiac fibrosis, and that its DR-region represents a potential therapeutic target.
Efferocytosis (ER) plays a crucial role in the programmed clearance of dead cells, a process that is mediated by phagocytic immune cells. However, further exploration is needed to determine the full extent of its impact on the progression of pancreatic ductal adenocarcinoma (PDAC), particularly through interactions among tumor cells, stromal cells, and immune cells within the tumor microenvironment (TME).
In this study, we comprehensively analyzed the Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) database, as well as additional databases from multiple bioinformatics websites, utilizing 167 ER features derived from the integration of single-cell RNA sequencing (scRNA-seq) and bulk transcriptomic data. A set of 14 ER-associated prognostic signatures, referred to as the “14-gene panel” genes, was identified based on overall survival (OS)/disease-free survival (DFS) data, Pearson correlation coefficients, and multivariate Cox regression analyses. The model pathways enriched by the four-gene combination represented by “LEAF” and the 14-gene combination represented by the “14-gene panel” presented a high degree of similarity, including among the adhesion, mitotic, G2/M checkpoint, and epithelial‒mesenchymal transition (EMT) signaling pathways. Least absolute shrinkage and selection operator (LASSO) regression was subsequently employed to construct an ER risk scoring system using deep learning, based on the following formula: LGALS3, EMP1, ASPH, and FNDC3B, collectively termed the “LEAF” panel. Additionally, random survival forest (RSF) algorithms facilitated the identification of a key panel of genes, designated “LEAP” genes, including LGALS3, EREG, ASPH, and PLS3; three of which genes (ASPH, LGALS3, and EREG) were identified as key factors influencing the behaviors of PDAC tumors, tumor-associated stroma, and macrophages. Finally, we utilized experimental methods, including Boyden chamber analyses, immunohistochemical staining, and cell cycle analyses, to demonstrate that interference with ASPH suppresses the malignant properties of tumors, including proliferation and migration. Multiplex immunofluorescence staining was employed to identify EREG as highly relevant to the M2 macrophage subpopulation.
Our findings underscore the importance of considering a novel prognostic signature comprising 14 ER genes in the context of the TME when investigating the biology of PDAC. Future studies may explore how modulating these interactions could lead to novel therapeutic opportunities.
Hepatocellular carcinoma (HCC) is the leading cause of cancer-related mortality worldwide. Despite advances in therapeutic approaches, the lack of effective biomarkers continues to limit early detection and prognostic evaluation. Pseudogenes, once considered nonfunctional, have emerged as regulators of biological processes in tumors and as potential biomarkers. This study aimed to identify and validate BMS1 Pseudogene 8 (BMS1P8) as a liver-specific, clinically relevant diagnostic and prognostic biomarker in HCC.
A comprehensive survey of pseudogene expression across different stages of liver disease was performed and validated using clinical HCC samples. Correlation, enrichment, and competing endogenous RNA (ceRNA) analyses integrating matched microRNA (miRNA)-seq and mRNA-seq were used to explore the functional networks surrounding BMS1P8. Public RNA-seq datasets (GSE114564, The Cancer Genome Atlas-Liver Hepatocellular Carcinoma (TCGA_LIHC)) were used to delineate differentially expressed pseudogenes, and 98 paired tumor and non-tumor tissues were assessed using quantitative reverse transcription polymerase chain reaction. Diagnostic and prognostic performances were evaluated using receiver operating characteristic curves and Kaplan–Meier statistics.
BMS1P8 was markedly upregulated in HCC and was overexpressed in 25 other cancer types. Receiver operating characteristics analysis yielded an area under the curve of 0.81, underscoring the diagnostic utility. High BMS1P8 expression and enrichment of cell cycle pathways were associated with poor survival. ceRNA screening revealed an inverse BMS1P8–miR-30c-2-3p correlation and concordant NME/NM23 nucleoside diphosphate kinase 6 (NME6) upregulation, with the BMS1P8/miR-30c-2-3p/NME6 triad further stratifying patient outcomes.
Our findings highlight BMS1P8 as a novel liver-specific biomarker with substantial diagnostic and prognostic value in HCC. Its diagnostic utility suggests its potential application in early detection and personalized treatment strategies, contributing to improved patient outcomes.
Ageing is a progressive functional decline in health conditions and a risk factor for many chronic diseases. To address the elevated burden of age-related pathologies, the ageing process has been extensively studied over the past decades, and yet the underlying mechanisms remain to be fully understood. One of the prominent features of ageing is cellular senescence, a special form of durable cell-cycle arrest. While senescent cells release the senescence-associated secretory phenotype (SASP) molecules that recruit immune cells to facilitate the clearance of senescent cells, senescence is also indispensable for many essential physiological functions. However, a ‘chronic’ nature of senescence arises due to immune deficiencies and persists during ageing. Immunosenescence, the ageing of immune cells, is the underlying key driving the pathological burdens of senescence, leading to systemic ageing as demonstrated by animal studies. Thymic regeneration has been shown by several studies to be a potential anti-ageing intervention, restoring immunity as well as reversing immunosenescence and ageing. The specific targeting of senescent cells by senolytic and/or senomorphic drugs is also promising but needs to be dealt with caution to protect the essential physiological roles of senescence. A deeper understanding of the biological origins of immunosenescence is crucial for unveiling the potential root cause of ageing.
Methamphetamine (METH) addiction is a global concern due to its severe impact on public health, including heightened aggression and neurotoxic effects. Genetic and epigenetic factors, particularly involving the SLC6A4 and COMT genes, are implicated in individual vulnerability to METH addiction. Thus, understanding the molecular mechanisms involved is crucial for developing targeted prevention and treatment strategies.
A systematic literature review was conducted following the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA) guidelines. Six major databases (MEDLINE/PubMed, Scopus, ScienceDirect, ResearchGate, Web of Science, Google Scholar) and Spanish-language platforms (Dialnet, Redalyc, CSIC, RECyT) were searched for studies published in English, Spanish, and Portuguese over the last 40 years. The inclusion criteria encompassed original research focusing on genetic and/or epigenetic determinants of METH addiction, with particular emphasis on the SLC6A4 and COMT genes. Studies focusing on substances other than METH, non-human subjects, or those that did not meet the language or temporal restrictions were excluded. Data on genetic variants, epigenetic alterations (e.g., DNA methylation, histone modifications), and relevant behavioral outcomes were extracted.
From an initial 600 articles, 25 studies met the inclusion criteria and were included in the qualitative synthesis. Polymorphisms in SLC6A4 (e.g., 5-HTTLPR) were associated with an increased risk of METH addiction (odds ratio (OR) = 2.31, 95% confidence interval (CI): 1.45–3.68; p = 0.001); meanwhile, variations in COMT (Val158Met) were linked to both susceptibility and executive function deficits. Epigenetic modifications—most notably DNA methylation in SLC6A4 and COMT—also emerged as important contributors to addiction pathways, potentially influencing dopamine and serotonin regulation. Gene-environment interactions, including factors such as childhood trauma and socioeconomic status, were found to modulate genetic predispositions, suggesting a multifaceted etiology for METH dependence.
Both genetic polymorphisms and epigenetic alterations play a critical role in METH addiction vulnerability. The reviewed evidence highlights the need for more comprehensive, regionally diverse studies and integrative approaches that combine genetics, neurobiology, and psychosocial factors. Such strategies could inform personalized prevention and treatment interventions, improving patient outcomes and mitigating the global burden of METH addiction.