Objectives Amiodarone remains an important antiarrhythmic drug, but cumulative exposure can cause pulmonary, hepatic, renal, thyroid, and reproductive toxicity. This systematic review critically compares interventions tested to prevent or attenuate amiodarone-induced injury, evaluates evidence strength and replication, and integrates the implicated molecular pathways. Methods PubMed, MEDLINE, Scopus, and Web of Science were searched from inception to July 2025 for English-language in vitro, animal, and human studies of protective interventions against amiodarone toxicity. Animal studies were evaluated using SYRCLE; in vitro studies using an adapted OHAT framework; and the human observational study using the applicable JBI checklist. Because of substantial heterogeneity, findings were synthesized narratively by organ, mechanism, replication, and translational readiness. Database-specific search strategies are reported in Supplementary Table S1. Disagreements were resolved by consensus; agreement was not quantified. Key findings Thirty-five reports were included. The evidence was overwhelmingly preclinical; only one observational human report was identified and no randomized clinical trial was found. Pulmonary models predominated. Vitamin E had the broadest replication, while curcumin, silymarin, l-carnitine, and grape-seed preparations were evaluated in more than one report or complementary model. Protection converged on attenuation of lipid peroxidation and inflammatory signaling, preservation of endogenous antioxidants and mitochondrial function, and suppression of apoptosis or TGF-β/Smad-associated fibrosis. Most studies had unclear risk of selection or performance bias. Conclusion Current evidence establishes biological plausibility but not clinical effectiveness. No protective adjunct can presently be recommended for routine use with amiodarone. Independent replication, clinically relevant exposure schedules, pharmacokinetic-interaction testing, and rigorously designed human studies are required.
Polycystic ovary syndrome (PCOS) is a multifactorial endocrine-metabolic and reproductive disorder affecting approximately 10-13% of women globally, although prevalence varies according to the diagnostic criteria and population studied. Emerging evidence identifies adipose tissue as an important endocrine and immune mediator of PCOS pathophysiology. In women with PCOS, adipose depots undergo maladaptive remodeling characterized by adipocyte hypertrophy, extracellular matrix (ECM) fibrosis, impaired angiogenesis, and immune cell infiltration. These changes disrupt metabolic flexibility, promote chronic low-grade inflammation, and alter systemic energy regulation. A critical component of this pathology is the crosstalk between adipokines and cytokines, which serves as a mechanistic bridge between metabolic disturbances and reproductive impairment. Dysregulation of leptin, adiponectin, chemerin, visfatin, and resistin, coupled with overexpression of pro-inflammatory cytokines such as tumour necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β, perpetuates insulin resistance (IR), hyperinsulinemia, and oxidative stress (OS). These interactions directly impair ovarian folliculogenesis, granulosa-theca cell function, and hypothalamic-pituitary-gonadal (HPG) signaling, thereby contributing to anovulation, hyperandrogenism, and subfertility. This narrative review critically synthesizes histopathological, molecular, and translational evidence linking pathological adipose remodeling to PCOS. It further explores the potential of adipokines and cytokines as biomarkers and discusses emerging therapeutic strategies to restore adipose homeostasis. Understanding adipose-immune-reproductive interactions offers new opportunities for precision medicine approaches, with the potential to transform PCOS management from symptomatic control to long-term disease modification.
Orosomucoid-like (ORMDL) proteins are a component of the serine palmitoyltransferase complex and a negative regulator of ceramide synthesis. Among these ORMDL3 in addition to its role in lipid homeostasis, plays a key role in inflammation, endoplasmic reticulum stress, and immune regulation. Single nucleotide polymorphisms in the 17q12–q21 locus, which harbors ORMDL3, have been associated with multiple autoimmune diseases, including type 1 diabetes (T1D). T1D results from dysregulation of both β-cells and immune cells; yet the role of ORMDL3 in disease pathogenesis remains unclear. Here, we deleted Ormdl3 specifically in β-cells of the T1D-prone non-obese diabetic mice. Our analyses demonstrate that loss of ORMDL3 in β-cells prior to the initiation of immune infiltration does not affect diabetes incidence, islet morphology, circulating insulin/proinsulin levels, or islet inflammation, and immune cell profiles. These findings indicate that ORMDL3 function in β-cells is dispensable in this preclinical model of T1D and underscore the need to investigate the role of ORMDL3 in immune cells, as well as the contribution of other genes within the 17q12–q21 locus to T1D susceptibility.
This study aims to identify and analyze published studies in which anti-inflammatory or immunoregulatory compounds have been investigated and their effects evaluated using Biomphalaria glabrata as an experimental model. B. glabrata, a mollusk highly susceptible to Schistosoma mansoni, is increasingly being considered a promising model for studying inflammatory processes and screening potential therapeutic agents. Inflammation, although a natural component of the immune response, can become chronic and contribute to the development of various diseases. In this mollusk, hemocytes act as key immune cells, responding to parasitic infections through mechanisms functionally analogous to inflammation in vertebrates. Substances such as hydrocortisone and Tenofovir Disoproxil Fumarate (TDF) have demonstrated the ability to alter immune responses in B. glabrata, either by suppressing hemocyte function, reducing oxidative stress, or regulating immune signaling pathways. Due to its low maintenance cost, ease of handling, and rapid life cycle, B. glabrata represents a valuable invertebrate model for early-stage screening of immunomodulatory and potentially therapeutic compounds. Overall, this work supports the use of B. glabrata as a viable alternative model in research, contributing to the development of novel anti-inflammatory strategies and reducing the need for early testing in mammalian systems.
Background Alzheimer's disease (AD) is the most common neurodegenerative disease worldwide, imposing a substantial economic burden. As the population ages and cases rise, interest in early identification, prevention, and treatment has intensified. After establishing that pathology involves neurofibrillary tangles, neuroinflammation, and neuronal loss, interventions have been tested in laboratory animals to slow disease progression. One commonly tested intervention is physical exercise, an economical approach. Laboratory animal models of AD aim to demonstrate how different types and intensities of exercise affect disease pathology. This review aims to categorize and elucidate the mechanisms and their outcomes. Materials and methods Original manuscripts concerning the impact of exercise, with or without medicinal intervention, on Alzheimer's dementia progression modeled in laboratory animals from 2020 onwards were included. The routes through which exercise exerts its impact at molecular, behavioral, and histomorphological levels were categorized. Results The most frequent modeling approach was Aβ injection into the hippocampus of Wistar rats, followed by Streptozocin and other chemicals, and transgenic models. Exercise mainly consisted of treadmill or swimming. Interventions were categorized into short-, medium-, and long-term protocols, ranging from 3 to 24 weeks, with most at 4 weeks, 30–60 min per session, 5 days/week. Molecular tests primarily measured Reactive Oxygen Species (ROS), while the most common behavioral test was the Morris Water Maze (MWM), assessing learning and memory. Histopathological assessments focused on β-Amyloid plaque formation, dark cells, and glial activation. Most studies agreed on AD modeling methods, but exercise protocols varied in intensity, duration, and type. The combined use of molecular, behavioral, and histological tests to assess intervention effects was consistent. Conclusion Physical exercise has been linked to lower oxidative stress and neuroinflammation, enhanced cognitive function, and fewer pathological changes in animal models of AD. However, variability in disease models and exercise protocols makes it difficult to identify the best exercise approach.
Gastric cancer (GC) remains one of the leading causes of cancer-related mortality worldwide, largely due to its high metastatic potential and poor prognosis at advanced stages. Epithelial–mesenchymal transition (EMT) is a pivotal biological process that facilitates tumor invasion, dissemination, and therapeutic resistance in GC. In recent years, long non-coding RNAs (lncRNAs) have emerged as critical regulators of gene expression, orchestrating multiple aspects of cancer progression through epigenetic, transcriptional, and post-transcriptional mechanisms. This review provides a comprehensive overview of the current understanding of lncRNA-mediated regulation of EMT and metastasis in gastric cancer. We summarize how lncRNAs function as epigenetic modulators, competing endogenous RNAs (ceRNAs), and molecular scaffolds to regulate EMT-associated transcription factors, signaling pathways, and chromatin dynamics. In particular, key oncogenic pathways such as Wnt/β-catenin, PI3K/AKT, STAT3, and NF-κB are highlighted as major downstream targets of lncRNA activity. Furthermore, we discuss the involvement of lncRNAs in tumor microenvironment remodeling, hypoxia response, angiogenesis, immune regulation, and extracellular matrix reorganization, all of which contribute to metastatic progression. Importantly, dysregulated lncRNA expression is closely associated with clinicopathological features and patient outcomes, underscoring their potential as diagnostic and prognostic biomarkers. In addition, the therapeutic targeting of lncRNAs represents a promising strategy for overcoming metastasis and treatment resistance in gastric cancer. Overall, this review integrates recent advances in the field and provides insights into the complex regulatory networks governed by lncRNAs, highlighting their translational potential in improving GC management.
Drug development is a complex, time-consuming, and costly process, making drug repurposing an important strategy for identifying new therapeutic applications for existing drugs. Computational drug-disease association (DDA) prediction has therefore attracted increasing attention in recent years. Although many existing methods incorporate similarity information and graph neural networks for DDA prediction, effectively capturing both local structural patterns and global dependencies, as well as modeling complex interactions between drugs and diseases, remains challenging. To address these limitations, we propose Drug-disease association prediction model (KNN, GIN, Graph Transformer and Cross-attention, KGTC-DDA), a novel framework that combines Graph Isomorphism Networks (GIN), Graph Transformers (GT), and cross-attention mechanisms for drug-disease association prediction. Specifically, multiple biological similarities, including drug chemical structure similarity, disease phenotype similarity, and Gaussian interaction profile (GIP) kernel similarity, are integrated to construct comprehensive drug and disease similarity graphs. GIN and Graph Transformer modules are then employed in parallel to simultaneously learn local topological features and global contextual representations from the similarity networks. Furthermore, a cross-attention mechanism is introduced to capture high-order interdependencies between drug and disease embeddings, thereby enhancing heterogeneous feature interaction and representation learning. Extensive experiments on three benchmark datasets under 10-times 10-fold cross-validation demonstrate that KGTC-DDA achieves superior predictive performance compared with several state-of-the-art methods in terms of AUROC and AUPR. In addition, experiments on unseen disease prediction further validate the robustness and generalization capability of the proposed framework. These results indicate that KGTC-DDA provides an effective and promising approach for computational drug repositioning and drug-disease association prediction.
Ice-binding proteins (IBPs) are functional molecules that enable organisms to survive severe cold by binding specifically to ice crystals and suppressing their growth and recrystallisation. Although their primary role has traditionally been viewed as antifreeze activity, recent studies have suggested additional cell-protective functions, including possible effects on membrane stability. Heterologous expression of the fish type III IBP NfeIBP in Caenorhabditis elegans has previously been shown to increase tolerance to cold and freezing stress. However, the understanding of the intracellular localisation of IBPs in vivo and their effects on gene expression remains limited. Here, we examined the intracellular localisation and associated cellular effects of the fish-derived IBP variant NfeIBP6(A20I), which is known to confer strong cold tolerance in C. elegans. An NfeIBP6(A20I)–wrmScarlet fusion protein expressed in body wall muscle cells was detected in both cytoplasmic and nuclear regions and partially overlapped with the nuclear marker NLSsfGFP, indicating its presence within nuclear compartment. Expression of NfeIBP6(A20I) did not cause detectable abnormalities in the physiological traits examined. Transcriptomic analysis further revealed biased changes in gene expression, with substantially more downregulated genes than upregulated genes. The downregulated genes were significantly enriched in biological processes related to cuticle development and immune response. These findings indicate that NfeIBP6(A20I) exhibits previously unrecognised intracellular behaviour in C. elegans and suggest that the functional scope of IBPs may extend beyond their conventional role in ice binding.
GLUT2 (Slc2a2) is a key glucose transporter in pancreatic β-cells, and its reduced expression is closely linked to defective glucose-stimulated insulin secretion (GSIS) and diabetes. We previously reported that pancreatic β-cell–specific nardilysin (NRDC)-deficient mice (BetaKO) exhibit a severe diabetic phenotype with defective GSIS and reduced Slc2a2 expression in islets. However, because BetaKO mice also showed reduced MafA, a key upstream regulator of Slc2a2, along with an increased α-cell/β-cell ratio and other secondary changes that could influence GLUT2 levels, the mechanism by which NRDC regulates Slc2a2 transcription remained unclear. Here, we demonstrate that NRDC controls Slc2a2 expression in a β-cell-autonomous and MafA-independent manner. By integrating publicly available ATAC-seq and ChIP-seq datasets, we identified four active enhancer regions around the murine Slc2a2 locus, two of which are evolutionarily conserved in human islets. Luciferase assays in MIN6 β-cells revealed that NRDC selectively controls the activity of a conserved enhancer located 39 kb downstream of the Slc2a2 transcription start site. Chromatin immunoprecipitation (ChIP) and re-ChIP assays further revealed that, in MIN6 cells, NRDC occupies this enhancer and is required for efficient recruitment of ISLET1, a transcription factor upstream of Slc2a2. These findings suggest that NRDC contributes to Slc2a2 regulation in addition to MafA, highlighting multifaceted roles of NRDC in pancreatic β-cell gene regulation.
Background Intellectual disabilities (IDs) are part of neurodevelopmental disorders (NDDs) and are genetically heterogeneous conditions characterized by impairments in cognition, learning, and adaptive functioning. Despite advances in gene discovery, many individuals, particularly those from understudied populations, remain without a molecular diagnosis. Recent reports implicate CCDC82 (HGNC: 26282) as an autosomal recessive ID gene, although the phenotypic spectrum and biological context remain incompletely defined. Methods Exome sequencing (ES) was performed in a consanguineous Pakistani family (PKMR06A) with four affected individuals presenting with moderate to severe ID. Variant segregation was confirmed by Sanger sequencing. In silico analyses, including pathogenicity prediction, protein structural modeling, and domain intolerance assessment, were used to evaluate the functional consequences of the identified variant. Spatiotemporal gene expression patterns were examined using bulk and single-cell human brain transcriptomic datasets. Results Clinically, affected individuals of family PKMR06A presented with early childhood global developmental delay, speech delay, hypotonia, gait abnormalities, spasticity, and mild facial dysmorphism. Genetic screening revealed a recurrent rare homozygous frameshift variant in CCDC82 (NM_024725.4): c.373del; p.(Asp125Ilefs*6), segregating with disease in all available affected individuals of the family. The identified c.373del variant was absent from the gnomAD database and was classified as pathogenic (PVS1, PM2, and PP1) based on ACMG/AMP criteria. The c.373del variant is predicted to introduce a premature termination codon, p.(Asp125Ilefs*6), leading to deletion of essential coiled-coil domains from the encoded protein, supporting a loss-of-function mechanism. In silico, transcriptomic analyses demonstrated preferential CCDC82 expression during prenatal human brain development, providing developmental context for the neurodevelopmental phenotype associated with the identified truncating variant. Conclusions This study expands the mutational landscape of CCDC82 and provides additional clinical and molecular evidence supporting its role in autosomal recessive NDD. The findings reinforce the importance of CCDC82 in human neurodevelopment and highlight the value of genomic investigation in underrepresented populations.
Neonatal necrotizing enterocolitisis a fatal gastrointestinal emergency affecting preterm infants, which is linked to destruction of intestinal barrier. The structural overexpression and negative regulation of Toll like receptor 4, a key receptor of lipopolysaccharidefrom Gram-negative bacteria in preterm intestinal epithelial cells, constitute a core hub connecting flora dysbiosis and barrier breakdown. When TLR4 neutrally binds with LPS, it signals the activation of the NF-κB-mediated inflammatory cascade through the MyD88/TRIF pathway which in turn induces apoptosis, necroptosis, and tight junction protein degradation in intestinal epithelial cells. degradation of tight junction protein of intestinal epithelial cells. The body naturally possesses protective mechanisms such as competitive receptors, intracellular negative regulatory networks, and microbial metabolites, which can be the target of drugs. Methods for the protection of the intestinal barrier by fine-tuning TLR4 signaling activity include small-molecule drugs, nano-delivery systems and nutritional interventions. Nonetheless, clinical translation remains hindered by the issues of TLR4 gene polymorphism, gestational age-dependent regulation, and immune balance maintenance. In the future, efforts should concentrate on the regulatory mechanisms of small molecule drugs and naturally occurring foodborne peptides. The goal is to combine the precision offered by genetic screening with a specific delivery system. This will allow for the targeted prevention and control of TLR4 signal systems. Ultimately, this will lead to a breakthrough in tackling the challenges associated with NEC treatment.
Organelles establish dynamic contacts to facilitate inter-organelle communication. Although their structures and dynamics are commonly observed by light microscopy, the spatial precision of organelle proximity is restricted by optical diffraction limit. While super-resolution microscopy has significantly advanced the visualization of subcellular ultrastructure, optimizing imaging parameters and robust downstream analysis remains a key challenge. Here we present a practical workflow for live-cell super-resolution imaging and quantitative analysis of organelle spatial proximity. By combining super-resolution microscopy and machine learning-driven batch image processing, this method enables accurate estimation of organelle morphology and juxtapositions. Notably, this workflow is broadly adaptable to different subcellular structures, labeling strategies and imaging conditions. It is designed to be accessible to most cell biology laboratories without requiring large-scale training datasets or extensive computational resources.
Osteosarcoma is an aggressive bone malignancy characterized by early metastasis, chemoresistance, and poor prognosis, particularly in recurrent or metastatic cases. The long non-coding RNA PVT1 has been implicated as an oncogene in various cancers, but its therapeutic potential in osteosarcoma remains underexplored. This study investigates the feasibility of targeting PVT1 using lipid nanoparticle-encapsulated siRNA (LNP-siPVT1) as a standalone or combination therapy for osteosarcoma. PVT1 expression was found to be significantly upregulated in osteosarcoma cell lines (U2OS, Saos-2, MG-63), patient tissues, and public datasets (GSE126209 and GSE309091). Tumor-targeted LNPs encapsulating siPVT1 were synthesized via microfluidic mixing and exhibited a mean diameter of ∼100 nm, favorable encapsulation efficiency, and accelerated siRNA release under acidic pH (5.5). In vitro, LNP-siPVT1 effectively suppressed PVT1 expression, reduced cell viability (IC50 = 29.30 nM), and induced apoptosis in MG-63 cells; moreover, combining LNP-siPVT1 with doxorubicin (DOX) or sorafenib produced synergistic cytotoxic and pro-apoptotic effects. In a humanized immune system mouse model bearing MG-63 xenografts, LNP-siPVT1 plus DOX significantly inhibited tumor growth compared to either monotherapy, without causing overt toxicity or body weight loss, as confirmed by histology and serum biochemistry. Collectively, PVT1 serves as a prognostic biomarker and a promising therapeutic target in osteosarcoma, and LNP-mediated delivery of siPVT1, especially in combination with conventional chemotherapeutics such as DOX, represents an effective and safe strategy to enhance anti-osteosarcoma efficacy, supporting further clinical translation of LNP-siPVT1-based therapy.
Objective To develop a novel experimental autoimmune prostatitis (EAP) model in rats using a prostate-specific antigen (PSA) mixture and to determine the optimal dose by evaluating the dose-dependent effects. Methods Male SD rats were divided into control (saline), low-dose (0.1 mL PSA mixture), and high-dose (0.2 mL PSA mixture) groups. After 7 days of daily injections, general status, body weight, and prostate histopathology were assessed. Serum immunoglobulins IgA, IgG, IgM and cytokines IL-1β, IL-10 were measured by ELISA, while tissue TNF-α and CRP were detected by immunohistochemistry. Testosterone, androgen, and liver/kidney function indicators were also analyzed. Results Compared with the control group, both model groups exhibited successful EAP induction, as evidenced by significant weight loss, disrupted prostate glandular architecture, and severe inflammatory cell infiltration. These pathological changes were consistently more pronounced in the high-dose group. Immunological analysis revealed that model groups had significantly elevated serum immunoglobulins and pro-inflammatory factors, while levels of the anti-inflammatory factor IL-10 and local testosterone and androgens were markedly decreased. Importantly, no significant damage was detected in major organ biochemical indicators or histopathology, confirming the good biosafety of the modeling method. Conclusion This study successfully developed a novel EAP modeling method based on a PSA antigen mixture. The model effectively mimics the typical pathological and immune characteristics of chronic prostatitis and exhibits a clear dose-dependent response, providing an optimized and reliable experimental tool for subsequent research into the pathogenesis of chronic prostatitis and the evaluation of drug efficacy.
Central nervous system (CNS)-associated diseases such as neurological (including cerebral ischemia-reperfusion injury and stroke, Parkinson's disease, Alzheimer's disease, traumatic brain injury, and spinal cord injury) and psychological (including depression, anxiety, attention deficit hyperactivity disorder, and dementia) diseases are known as serious problems for public health worldwide. These diseases contribute to the occurrence of several disabilities and even death in affected individuals, and also decrease their quality of life. Therefore, discovering effective complementary treatments is essential. Recently, baicalin, a multifunctional natural product, has attracted much attention because of its therapeutic potentials in various diseases. Moreover, it has been shown that this agent can regulate key cellular and molecular processes in human diseases. In the case of CNS-related diseases, this review article aimed to summarize and discuss the available data from in vivo and in vitro investigations on the therapeutic application of baicalin, based on underlying mechanisms.
Articular cartilage injury is a common clinical condition in orthopedics. The avascular and aneural nature of the tissue results in extremely poor self-healing ability, which seriously impairs patients' motor function and quality of life. Current clinical treatment methods such as bone marrow stimulation, chondrocyte transplantation, and artificial joint replacement generally have limitations including the repaired tissue being primarily fibrocartilage, poor integration with host cartilage, and the need for secondary surgery, making them difficult to meet clinical needs. Hydrogels have become promising tissue engineering scaffolds for mimicking the natural cartilage extracellular matrix (ECM) due to their high water content, three-dimensional porous structure, and excellent biocompatibility. However, their defects such as insufficient mechanical strength, lack of biological activity, and antibacterial properties limit their clinical application and translation. Zinc oxide nanoparticles (ZnO NPs) are multifunctional bioactive materials that can enhance hydrogel mechanical stability and provide antibacterial, anti-inflammatory, and chondrogenic benefits through controlled Zn2+ release. This review summarizes the physicochemical properties and biological functions of ZnO NPs, discusses strategies for constructing ZnO NP-enhanced biomimetic hydrogels, and reviews current in vitro and in vivo evidence supporting their application in cartilage repair. It also highlights key challenges, including biosafety, concentration control, and translational feasibility, providing guidance for the rational design and clinical development of ZnO-based hydrogel scaffolds. Recent studies have explored hydrogel-based strategies to address this limitation by providing a microenvironment that favors hyaline cartilage regeneration over fibrocartilage formation.
Human-induced pluripotent stem cells (iPSCs) hold considerable potential for generating motor neurons (MNs), offering new avenues for disease modeling and regenerative medicine. To elucidate the molecular mechanisms underlying iPSC differentiation into MNs, we conducted an integrated bioinformatics analysis of transcriptomic data from the mature Day 28 stage of a publicly available 28-day iPSC differentiation dataset. Protein-protein interaction (PPI) networks were constructed using STRING and visualized in Cytoscape, while CytoHubba identified 15 hub genes, including BUB1, TOP2A, AURKB, CCNA2, and TP53, which are primarily associated with cell cycle regulation, mitotic progression, and genomic stability. Gene Ontology (GO) enrichment analysis identified biological functions related to chromosomal organization, cytoskeletal remodeling, metabolic activity, and translational regulation, whereas Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis highlighted cell cycle, DNA replication, cellular senescence, p53 signaling, and oocyte meiosis. CytoCluster analysis identified distinct subnetwork modules linked to neuronal differentiation. Promoter motif analysis via MEME uncovered conserved transcription factor binding sites, whereas miRNA-mRNA interaction prediction using the psRNATarget database identified 5316 potential regulatory pairs that may fine-tune gene expression during MN maturation. Codon usage analysis of 163 genes indicated moderate to high translational optimization, likely influenced by both mutational bias and selective codon preferences, with GC3 content strongly affecting codon choice. These patterns suggest potential mechanisms that may promote efficient protein synthesis during MN differentiation; however, experimental validation is required. This systems-level bioinformatics framework integrates multiple computational analyses to characterize transcriptional, post-transcriptional, and translational regulatory mechanisms associated with human MN differentiation. However, as these results are derived from bioinformatics analyses, they warrant further experimental validation and have the potential to advance our understanding of the molecular interactions and signaling pathways that regulate MN differentiation and maturation. These findings provide a computational framework for exploring regulatory mechanisms of motor neuron differentiation and prioritizing candidate genes for future experimental validation.
