Fibrotic diseases encompass a range of pathological conditions characterized by the abnormal growth of connective tissue, involving various cell types and intricate signaling pathways. Central to the onset and development of fibrosis are macrophages and fibroblasts, whose interactions are a pivotal area of investigation. Macrophages facilitate the activation, growth, and collagen production of fibroblasts, doing so either directly or indirectly through the release of cytokines, chemokines, and growth factors. Conversely, fibroblasts boost macrophage activity and intensify local inflammatory responses by secreting cytokines and matrix proteins associated with fibrosis. Throughout the different phases of fibrosis, these two cell types communicate via cytokines and signaling pathways, thereby sustaining the pathological condition. This review emphasizes the interplay between macrophages and fibroblasts and their contributions to fibrosis in the lungs, liver, kidneys, and other organs. Furthermore, it delves into potential therapeutic targets within these interactions, with the aim of shedding light on future clinical research and treatment approaches for fibrotic diseases.
The Akt/PKB (protein kinase B) is a major transducer of the phosphoinositide 3-kinase (PI3K) signaling axis, regulating key cellular processes such as growth, proliferation, apoptosis, survival, and migration in both normal and cancer cells. In normal cells, oncoproteins and tumor suppressor proteins within the Akt pathway exist in equilibrium. However, this equilibrium is disrupted in cancer cells due to activating mutations in oncoproteins and inactivating mutations in tumor suppressor proteins. This dysregulation drives tumor growth and progression, making the Akt pathway an attractive target for cancer therapies. A deeper understanding of the molecular mechanisms of the Akt signaling pathway is crucial for developing novel therapeutic agents targeting Akt and its downstream effectors for cancer treatment. This review discusses the role of Akt in cancer, current Akt-targeted agents, their limitations, and future trends.
Antibody-drug conjugates (ADCs) are an emerging class of cancer therapeutics comprised of a tumor-targeting antibody linked to a cytotoxic payload. Sacituzumab govitecan (SG or IMMU-132) is comprised of a trophoblast cell-surface antigen 2 (TROP-2)-directed antibody linked to the topoisomerase 1 inhibitor, SN-38. SG was designed to exploit the overexpression of TROP-2, observed in a variety of different epithelial cancers, to enhance tumor-selective cytotoxicity while minimizing damage to normal tissues. SG is approved for pretreated metastatic triple-negative breast cancer (mTNBC) and hormone receptor-positive human epidermal growth factor receptor 2 (HER2) negative breast cancer patients. While SG has shown significant clinical benefit, the objective response rate (ORR) observed with SG in pretreated mTNBC patients in the Phase I/II basket study was 33.3%, indicating a heterogeneous response profile to SG. This article explores the potential influence of autophagy, senescence, and the patient’s immune system on the treatment response.
The prevalence of male infertility attributed to oxidative stress (OS) is a growing concern globally. Traditional methods to treat male infertility have some limitations, including low efficacy and invasiveness. Additionally, assisted reproductive procedures, such as in vitro fertilization and intracytoplasmic sperm injection, are expensive and carry higher risks. These challenges underscore the need for innovative solutions. A multidisciplinary approach is imperative, drawing insights from fields such as reproductive biology, nanotechnology, and clinical research to effectively combat male infertility caused by OS. Recent advancements in nanobiotechnology provide a promising opportunity to tackle male infertility caused by OS. These advancements enable the design and development of nanoantioxidants (nanoAOXs) and drug delivery systems tailored to the male reproductive environment. This review highlights the recent progress in the rational design of nanomaterials, with a specific focus on nanoAOXs for managing male infertility associated with OS.
Ischemic stroke ranks among the leading global causes of death and disability, driven by intricate mechanisms such as neuronal injury, inflammation, and oxidative stress. Emerging as a pivotal player in ischemic stroke progression is ferroptosis—an iron-dependent form of regulated cell death. Its hallmarks—iron metabolic dysregulation and lipid peroxidation—trigger cell membrane disruption and irreversible neuronal damage. Beyond that, ferroptosis intensifies inflammation and compromises the blood–brain barrier (BBB), substantially increasing the impact of ischemic injury. Research indicates that modulating ferroptosis-related molecular pathways could significantly mitigate the pathological progression of ischemic stroke. Based on a systematic search of PubMed, Web of Science, Embase, and Cochrane Library databases (as of April 30, 2025), this review focuses on the progress of research on the mechanisms and treatments of ferroptosis in ischemic stroke over the past five years, aiming to investigate the underlying mechanisms, pathological roles, cross-disease associations, and targeted therapeutic strategies, to lay a theoretical foundation for the development of advanced therapies, and to outline the challenges and future directions of the field.
The important immunoregulatory roles of regulatory T cells (Tregs) include fostering tolerance to infections, controlling immune surveillance, and curtailing autoimmunity. Years of research have not only generated abundant knowledge in the field of Treg biology but also enabled the initial application of Tregs in cell therapy. However, most data in this field are obtained from laboratory animals and in vitro experiments. This review provides an updated summary and the latest understanding of Treg-targeting cell therapy. We introduce the unique traits of Tregs, review animal experiments and clinical trials on Treg injections, discuss limitations of Treg applications, and consider future perspectives on Treg-based therapies. Overall, the safety and potential efficacy of Tregs will broaden the scope of cell-based treatments.
Lung cancer, the leading cause of cancer-related mortality worldwide, poses considerable therapeutic challenges due to the varied responses to programmed death-1/programmed death-ligand 1 (PD-1/PD-L1) inhibitors. Emerging highlight the pivotal role of host-microbiome interactions in modulating antitumor immunity and influencing clinical outcomes. This review examines how the respiratory and gut microbiota contribute to the immunosuppressive tumor microenvironment through dysbiosis-induced T-cell exhaustion and regulatory cell activation, while certain commensals facilitate dendritic cell-mediated recruitment of cytotoxic T lymphocytes. Additionally, this review explores the molecular mechanisms by which microbial metabolites, such as short-chain fatty acids, influence myeloid-derived suppressor cells. Therapeutically, microbiota-modulation strategies—such as tailored probiotic formulations and precision fecal microbiota transplantation—offer potential to enhance immunotherapy efficacy. This review provides a foundation for microbiome-guided immunotherapy, advocating for biomarker-driven patient stratification and the use of engineered microbial consortia to counteract therapeutic resistance. These findings pave the way for the integration of microbiome science into next-generation precision oncology.
The role of macrophages has transcended the traditional binary framework of M1/M2 polarization, emerging as “tissue microenvironment engineers” that dynamically govern organismal homeostasis and disease progression. Under physiological conditions, they maintain balance through phagocytic clearance, metabolic regulation (e.g., lipid and iron metabolism), and tissue-specific functions (such as hepatic detoxification by Kupffer cells and intestinal microbiota sensing), all meticulously orchestrated by epigenetic mechanisms and neuro-immune crosstalk. In pathological states, their functional aberrations precipitate chronic inflammation, fibrosis, metabolic disorders, and neurodegenerative diseases. Notably, this plasticity is most pronounced within the tumor microenvironment (TME): tumor-associated macrophages (TAMs) polarize toward a protumoral phenotype under conditions of low pH and high reactive oxygen species (ROS). They promote angiogenesis via vascular endothelial growth factor (VEGF), suppress immunity through interleukin-10 (IL-10)/programmed death-ligand 1 (PD-L1), and facilitate tumor invasion by degrading the extracellular matrix, ultimately fostering an immune-evasive niche. Novel intervention strategies targeting TAMs in the TME have shown remarkable efficacy: CRISPR-Cas9 spatiotemporal editing corrects aberrant gene expression; pH/ROS-responsive nanoparticles reprogram TAMs to an antitumoral phenotype; chimeric antigen receptor-macrophage (CAR-M) 2.0 enhances antitumor immunity through programmed death-1 (PD-1) blockade and IL-12 secretion; and microbial metabolites like butyrate induce polarization toward an antitumor phenotype. Despite persisting challenges—including the functional compensation mechanisms between tissue-resident and monocyte-derived macrophages, and obstacles to clinical translation—the macrophage-centered strategy of “microenvironmental regulation via cellular engineering” still holds revolutionary promise for the treatment of tumors and other diseases.
Pericytes (PCs) are multifunctional mural cells embedded in the basement membrane of microvessels and play essential roles in the development and maintenance of the central nervous system. This review provides a comprehensive synthesis of the current knowledge on PC biology, tracing their trajectory from embryonic origins to specialized functions in the adult brain. During early brain development, PCs are recruited via platelet-derived growth factor B (PDGF-BB)/platelet-derived growth factor receptor beta (PDGFRβ) signaling and contribute to the formation of the blood–brain barrier (BBB), cortical architecture, and vascular stability. Their developmental plasticity is shaped by multiple embryonic origins and dynamic interactions with endothelial and neural precursor cells. In the adult central nervous system, PCs are central to maintaining BBB integrity, regulating cerebral blood flow, and modulating neurovascular coupling. They also participate in immune responses, metabolic waste clearance, and neuroprotection through the secretion of trophic factors and cytokines. Of particular interest is their emerging role in the expression of lipocalin-type prostaglandin D synthase (L-PGDS), which synthesizes prostaglandin D2—a molecule involved in sleep regulation, inflammation, and neurodegeneration. L-PGDS may also act as an amyloid β chaperone, implicating PCs in the pathology of Alzheimer’s disease and other neurodegenerative disorders. The regulatory mechanisms of L-PGDS expression involve nuclear factor kappa B and Notch–Hes signaling, as well as potential modulation via brain-derived neurotrophic factor/tropomyosin receptor kinase B/protein kinase C pathway. By integrating developmental, molecular, and pathophysiological perspectives, this review positions PCs as key cellular regulators of brain function and highlights their potential as therapeutic targets in cerebrovascular and neurodegenerative diseases.
Histone post-translational modifications (HPTMs) have emerged as crucial epigenetic regulators in urological malignancies, including prostate, bladder, and renal cell carcinomas. This review systematically examines four key modifications—lactylation, acetylation, methylation, and phosphorylation—and their roles in carcinogenesis. These dynamic modifications, mediated by “writers”, “erasers”, and “readers”, influence chromatin structure and gene expression, thereby driving oncogenic processes such as metabolic reprogramming, immune evasion, and treatment resistance. The newly discovered lactylation modification links cellular metabolism to epigenetic regulation through lactate-derived histone marks, particularly in clear cell renal cell carcinoma, where it activates oncogenic pathways. Acetylation modifications, regulated by histone acetyltransferases (HATs) and histone deacetylases (HDACs), modulate chromatin accessibility and are implicated in silencing cancer suppressors. Methylation patterns, controlled by histone lysine methyltransferases (KMTs) and histone lysine demethylases (KDMs), demonstrate dual roles in gene regulation, with specific marks either promoting or suppressing carcinogenesis. Finally, phosphorylation dynamics affect critical cellular processes such as cell cycle progression and DNA repair. This review underscores the therapeutic potential of targeting these modifications, as evidenced by promising results with HDAC and Enhancer of zeste homolog 2 (EZH2) inhibitors. However, challenges persist in clinical translation, including off-target effects and the complexity of the cancer microenvironment. Future research should utilize multi-omics approaches to elucidate modification crosstalk and develop precision therapies. Overall, this comprehensive analysis provides valuable insights into the epigenetic mechanisms underlying urological cancers and highlights remaining knowledge gaps and therapeutic opportunities in this rapidly evolving field.
Plant-derived treatments for skin inflammation are gaining increasing interest, driven by the growing demand for safer alternatives to conventional synthetic drugs. Curcuma longa L. (turmeric) is traditionally utilized in many Asian countries for various pharmacological applications. Although the inflammation-suppressing properties of turmeric rhizomes are well established, the bioactive potential of its leaves and pseudostems remains largely unexplored. This study investigates the effects of turmeric leaf and pseudostem extract (CLE) on tumor necrosis factor (TNF)-α/interferon (IFN)-γ-stimulated HaCaT keratinocytes (HK) and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear edema in a mouse model.
Cell viability and intracellular ROS levels in response to CLE were assessed. The potential of CLE to suppress inflammation was evaluated by monitoring the inhibition of signaling pathways and changes in cytokine/chemokine expression through Western blotting and real-time quantitative polymerase chain reaction (RT-qPCR) analyses. CLE was also examined for its impact on skin hydration and tight junction integrity. For in vivo analysis, an ear edema model was established using female BALB/c mice (7 weeks old).
CLE treatment led to a dose-dependent decline in intracellular ROS and enhanced cell viability of TNF-α/IFN-γ-stimulated HK. Treatment with CLE resulted in decreased transcription of epithelial-derived cytokines (thymic stromal lymphopoietin (TSLP), IL-25, IL-33), pro-inflammatory mediators (IL-6, IL-8, IL-13, TNF-α, IFN-γ, IL-1β), and chemokines (macrophage-derived chemokine (MDC), regulated on activation, normal T cells expressed and secreted (RANTES), thymus and activation-regulated chemokine (TARC)), along with inhibition of mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling proteins in stimulated HK. CLE improved expression of proteins associated with skin hydration and tight junctions, helping to preserve moisture balance and structural integrity. Moreover, CLE markedly reduced ear redness, swelling, and thickness in 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced mice, while alleviating histopathological changes, including inflammatory cell infiltration and dermal thickening. Additionally, CLE effectively diminished inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and pro-inflammatory cytokine expression in the ear tissues of edema-induced mice.
Collectively, CLE exhibited potential as a natural anti-inflammatory agent by attenuating oxidative stress, downregulating inflammatory mediators, enhancing skin barrier function in vitro, and reducing ear edema in vivo.
The intestinal microbiota, present in vast numbers within the human body, plays a pivotal role, with its composition and abundance varying significantly across individuals. This gut microbiota not only contributes to normal physiological development but also impacts the initiation, progression, resolution, and prognosis of various diseases. Recent studies have increasingly illuminated the connection between intestinal microbiota and pain, with a particular focus on the relationship between gut microbiota and neuropathic pain (NP). NP, an acute and chronic pain disorder arising from sensory nervous system injury, encompasses both peripheral and central neuropathic pain. Evidence suggests that intestinal microbiota influences NP occurrence and may modulate its severity. This review synthesizes current research findings on the microbiota-NP relationship, aiming to establish a theoretical foundation for future clinical investigations.
Claudins (CLDNs), key components of tight junctions, are dysregulated in various cancers. However, the roles and therapeutic potential of specific CLDN family members-particularly CLDN6, CLDN9, and CLDN10-in ovarian cancer (OC) remain incompletely defined. To address this gap, we conducted a comprehensive analysis of the CLDN family to identify novel diagnostic and prognostic biomarkers as well as potential therapeutic targets for OC.
Gene expression profiles and corresponding clinical data from The Cancer Genome Atlas ovarian cancer cohort (TCGA-OV) and two Gene Expression Omnibus (GEO) datasets (GSE18520, GSE26712) were analyzed. Differential expression of CLDN genes between OC and normal tissues was evaluated using R with appropriate bioinformatics packages (e.g., limma). Logistic regression models were employed to calculate odds ratios (ORs), and receiver operating characteristic (ROC) curves were generated across all datasets to identify consistently dysregulated CLDNs associated with OC. Prognostic hazard ratios (HRs) for these CLDNs were extracted from the Kaplan-Meier Plotter (KM Plotter) database and synthesized using a random-effects model to assess their associations with overall survival. Intersection analysis was performed to identify CLDNs exhibiting both significant differential expression and prognostic significance. Candidate targets underwent comprehensive validation, including single-cell RNA sequencing (scRNA-seq) to characterize cell-type-specific expression patterns. Notably, Key findings regarding CLDN6 were further validated by immunohistochemistry (IHC) on an independent tissue microarray (TMA), as well as functional assays in OC cell lines following siRNA-mediated knockdown. These included transwell invasion, wound healing (scratch) test, and measurements of mitochondrial depolarization, reactive oxygen species (ROS) accumulation, cell cycle arrest, and apoptosis.
CLDN6, CLDN9, and CLDN10 were consistently and significantly upregulated in OC compared to normal tissues across all datasets. Single-cell RNA sequencing revealed that CLDN6 and CLDN10 were predominantly expressed in malignant epithelial cell subsets, a pattern associated with aggressive tumor phenotypes. Meta-analysis of HRs showed that HR >1 in CLDN6 and HR <1 in CLDN10. Although CLDN10 is highly expressed in tumor cells, its hazard ratio (HR) is less than 1, and the underlying mechanism of this gene remains unclear. Experiments have confirmed that CLDN6 is closely associated with tumor invasion. Computational analysis, meta-analysis, and single-cell data collectively confirm that only CLDN6 is a clearly defined gene closely associated with tumor progression, a finding subsequently validated by experimental results. Notably, the combined signature comprising CLDN6, CLDN9, and CLDN10 exhibited superior diagnostic performance, with higher area under the curve (AUC) values in ROC analysis, compared to individual CLDNs or established OC biomarkers such as carbohydrate antigen 125 (CA125), human epididymis 4 (HE4), carcinoembryonic antigen (CEA), and alpha-fetoprotein (AFP). The signature also showed enhanced prognostic discrimination, as indicated by time-dependent ROC analysis. Protein overexpression of these targets was validated by IHC and Western blot. Functional assays further demonstrated that siRNA-mediated knockdown of CLDN6 significantly inhibited the proliferation of OC cells, promoted cell apoptosis, increased production of ROS, induced G1 phase arrest, inhibited cell invasion and migration in vitro. Furthermore, western blot analysis identified that knockdown of CLDN6 repressed the Wnt/β-catenin pathway. Nude mice experiments indicated that CLDN6 knockdown in OC cells dramatically suppresses the tumor growth and lung metastasis in vivo.
CLDN6, CLDN9, and CLDN10 are critically involved in the pathogenesis and progression of OC. A biomarker panel combining these three claudins demonstrates superior diagnostic and prognostic performance compared to individual markers and established clinical biomarkers such as CA125 and HE4. Notably, functional evidence indicates that CLDN6 plays a pivotal role in regulating malignant phenotypes, highlighting its potential as a novel therapeutic target. These findings collectively support the clinical utility of the CLDN6/9/10 axis as both a non-invasive biomarker signature and a promising avenue for targeted intervention in ovarian cancer.
Genes belonging to the adenylate cyclase (ADCY) family regulate various biological processes, including tumor metabolism, metastasis, angiogenesis, and immune escape. However, the functions of these genes in multiple cancers unclear.
This study analyzed the expression, prognostic value, correlation, mutation, and methylation patterns of ten genes belonging to the ADCY family across multiple cancers using multi-omics data. Additionally, the correlation between ADCY5 and immune cells, as well as the function of ADCY5 in multiple cancers were examined using single-cell data and spatial transcriptomic data.
Ten ADCY family genes were differentially expressed in most tumors and normal tissues, and their aberrant expression in multiple cancers significantly reduced patient survival. The expression level of ADCY5 was significantly correlated with the immune microenvironment. We also identified and validated the potential of ADCY5 as a potential biomarker for gastric cancer.
Our pan-cancer analysis nominates the ADCY family as a source of potential cancer biomarkers. We specifically validated ADCY5 in gastric cancer, establishing it as a promising prognostic biomarker with clinical and functional relevance, with significant implications for optimizing immunotherapy strategies and prognostic assessment in this malignancy.
Lymphangiogenesis and phenotypic transformation of endothelial cells are closely associated with the progression of renal interstitial fibrosis. Inflammatory injury triggered by mineralocorticoid receptor (MR) activation serves as the initial stimulus for lymphangiogenesis.
Thirty specific pathogen-free (SPF) male C57BL/6 mice were assigned to three groups randomly: the control group (CON), aldosterone-treated group (ALD group, in which aldosterone was infused at a rate of 0.75 μg/h via mini-osmotic pumps for 12 weeks), and esaxerenone-treated group (ESA group, administered at a dosage of 1 mg/kg/day via diet). The expression levels of lymphatic markers (lymphatic vessel endothelial hyaluronan receptor 1 (LYVE-1), vascular endothelial growth factor receptor 3 (VEGFR3), podoplanin, and VEGFC) were assessed using immunohistochemistry, immunofluorescence, and western blot analysis. Inflammatory injury markers (CD68, F4/80, IL-1β, TNF-α and TGF-β1) and endothelial–to–mesenchymal transition (EndMT, LYVE-1+ vimentin/α smooth muscle actin (α-SMA)+) were evaluated. In vitro, the effects of aldosterone on the migration, tube formation, and phenotypic transformation of human lymphatic endothelial cells (HLECs) in the presence of TGF-β1 or VEGFC were investigated.
In the ALD group, significant increases in lymphangiogenesis, macrophage infiltration, and the expression of TGF-β1, TNF-α, IL-1β and VEGFC were observed. Immunofluorescence double staining revealed that VEGFC was predominantly secreted by macrophages, and that lymphatic endothelial cells exhibited expression of vimentin and α-SMA. In vitro experiments demonstrated that aldosterone promoted HLECs migration and tube formation, as well as the activation of inflammatory cytokines and MR. Flow cytometry analysis indicated that HLECs underwent myofibroblastic transformation, which could be attenuated by MR blocker esaxerenone.
Aldosterone induces inflammatory injury, thereby promoting renal lymphangiogenesis and EndMT.
Peritoneal fibrosis is a significant complication arising from long-term peritoneal dialysis (PD), primarily due to the loss of peritoneal mesothelial cells (PMCs). Recent studies have implicated periostin (POSTN) in the progression of various fibrotic diseases; however, its specific role in PD-induced peritoneal fibrosis remains unclear. Sodium alginate (SA) microgels have emerged as promising carriers for cell encapsulation in tissue engineering and regenerative medicine. This study investigated the therapeutic potential of PMCs encapsulated in SA microgels (SA/PMC) for reducing PD-induced peritoneal fibrosis, with a focus on the modulation by the periostin/nuclear factor kappa-B (NF-κB)/CXCL8 signaling pathway.
Primary human peritoneal mesothelial cells (PHPMCs) were isolated from the PD effluent of patients. The effect of SA encapsulation on PMCs proliferation was evaluated using a Cell Counting Kit 8 (CCK-8) assay. The expression levels of POSTN, NF-κB p65, and CXCL8, as well as fibrosis markers, including α-smooth muscle actin (α-SMA), collagen I, transforming growth factor-β (TGF-β), and fibronectin, were evaluated in patients undergoing PD and a PD mouse model.
Patients undergoing PD for 1 year exhibited elevated levels of POSTN, NF-κB p65, CXCL8, and fibrosis markers compared with those undergoing PD for 1 week.
Consistent results from in vivo and in vitro models demonstrated that PD and hyperglycemic conditions upregulated the expression of POSTN, NF-κB p65, CXCL8, and profibrotic markers, leading to peritoneal thickening and fibrotic progression. Treatment with SA/PMC microgels ameliorated these effects. By modulating the POSTN/NF-κB/CXCL8 pathway and enhancing PMCs survival, SA/PMC microgels may have therapeutic potential in mitigating peritoneal fibrosis in PD patients.
Premature ovarian insufficiency (POI) is a condition marked by diminished ovarian function and reduced fertility, caused by the chemotherapy drug cyclophosphamide (CTX) used to treat gynecologic cancers. The abnormal inflammation of ovarian tissue induced by CTX represents a key factor that impairs follicular cells and disrupts fertility. Therefore, the present study aims to investigate the underlying mechanisms of CTX-induced abnormal ovarian inflammation and identify potential therapeutic agents.
RNA sequencing data derived from CTX-induced mouse ovarian tissues were first intersected with inflammation-related genes retrieved from the Gene Ontology (GO) database. This was followed by functional enrichments analysis and protein-protein interaction (PPI) analyses to identify target genes. Subsequently, the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) was screened to obtain corresponding candidate therapeutic agents. Finally, a CTX-induced mouse model was established to verify the therapeutic efficacy of the candidate drug and elucidates its underlying mechanisms.
A total of 25 candidate genes were identified, with interleukin 1β (IL1β) confirmed as the core gene. Subsequent screening resulted in the identification of Irisolidone as a potential therapeutic agent. The present study demonstrated that Irisolidone ameliorates CTX-induced follicular cell developmental impairment and improves fertility in mice with POI. Mechanistically, it was found that Irisolidone suppressed abnormal ovarian inflammation by inhibiting the CTX-disrupted nuclear factor kappa B (NFκB)/NOD-like receptor pyrin domain-containing 3 (NLRP3)/Caspase1 signaling pathway.
The present study demonstrates that Irisolidone can effectively alleviate CTX-induced POI by inhibiting abnormal inflammation. These findings suggest that Irisolidone holds promise as a novel therapeutic candidate for POI, thereby providing a potential new treatment strategy for clinical management of this condition.
The nucleotide “words” (k-mers) of the genome exhibit two essentially universal properties that follow probabilistically from the Conservation of Hartley-Shannon Information (CoHSI): (1) a Zipfian rank-ordered distribution of frequencies and (2) universal inverse symmetry. Here, we address the presence of these 2 properties in the transcriptome, a question of interest given the strong and specific structure/function constraints on RNAs, especially the protein-coding (CDS) sequences.
CDS and ncRNA (non-coding RNA) databases were accessed at e!Ensembl. For determination of a power-law, statistical tests of both necessity (linearity) and sufficiency (confidence that a power-law distribution could not be rejected) were applied. Compliance with inverse symmetry was assessed by linearity and residual standard error.
The CDS and non-coding RNAs for 53 species were analyzed separately and the data presented as short movies. The results were consistent for all species analyzed, and taking the bonobo (Pan paniscus) as a representative species, the following results were obtained. For the Zipfian distribution of k-mer frequencies, statistically robust tests of both necessity (adjusted R-squared of 0.9932 and p ≤ 2.2 × 10-16) and sufficiency were obtained for the CDS; for non-coding RNAs the test of necessity was robust (adjusted R-squared = 0.9982 and p ≤ 2.2 × 10-16). Perturbations of inverse symmetry were observed in both CDS (slope = 0.91, adjusted R2 = 0.77) and non-coding RNAs (slope = 1.02, adjusted R2 = 0.84). The disruption of inverse symmetry in the CDS affected particularly the 3- and 6-mers and was shown to be associated with codon (especially stop codon) frequency in the open reading frame.
Whereas the CoHSI-predicted Zipfian distribution of k-mer frequencies was observed in both the protein-coding and non-coding RNAs of 53 species, in contrast the compliance with inverse symmetry was weaker. This weakening of compliance was seen to a greater extent in the CDS than in the non-coding portions of the transcriptome and may be associated with the necessity to maintain the integrity of the reading frame in the CDS. These results illustrate the general principle that local perturbations of an overall CoHSI-guided equilibrium state of a biological system can provide insight into the underlying causes of such perturbations.
Heart failure (HF) remains a leading cause of morbidity and mortality worldwide. Although dapagliflozin, a selective sodium–glucose cotransporter 2 (SGLT2) inhibitor, has demonstrated significant cardiovascular benefits in large clinical trials, the underlying mechanisms beyond glucose lowering remain incompletely understood. Increasing evidence suggests that gut microbiota and its metabolites may contribute to HF progression through gut–heart axis interactions.
In this study, a total of 135 individuals with HF were recruited, comprising 84 patients treated with dapagliflozin (Y group) and 51 receiving conventional therapy (N group). Gut microbial communities were characterized through 16S rRNA gene sequencing to evaluate compositional structure, diversity metrics, and taxa differences between groups. Untargeted metabolomic profiling of plasma samples was conducted to identify significantly altered metabolites and enriched metabolic pathways. Furthermore, the interrelationships between gut bacterial taxa and circulating metabolites were systematically explored to delineate potential microbiome–metabolome interactions.
Dapagliflozin treatment significantly altered gut microbial composition (p < 0.05, permutational multivariate analysis of variance [PERMANOVA]), characterized by increased Prevotella, Akkermansia, Collinsella, and Fusobacterium, and reduced Bacteroides, Parabacteroides, Subdoligranulum, and Bifidobacterium in the dapagliflozin group, whereas control-enriched taxa included Lachnoclostridium and the Ruminococcus gauvreauii group. Fourteen plasma metabolites were differentially abundant between groups, including higher levels of O-phospho-L-threonine and epiandrosterone in the dapagliflozin group, while salicyluric acid and L- (+)-rhamnose were enriched in the control group. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated alterations in amino acid and one-carbon metabolism, as well as carbohydrate and steroid-related pathways. Correlation analysis revealed that Collinsella was positively associated with fludarabine phosphate (p < 0.05), whereas Akkermansia and Paraprevotella showed negative correlations with maslinic acid and phospho-L-valine, respectively (p < 0.01 to p < 0.001).
Dapagliflozin modulates gut microbiota composition and circulating metabolic signatures in HF patients, supporting a potential gut–heart axis mechanism contributing to its cardioprotective effects.
Cysteine and Glycine Rich Protein 1 (CSRP1) is a member of the cysteine-rich protein family, characterized by a unique double-zinc finger motif. It plays an important role in development and cellular differentiation. Aberrant expression of CSRP1 has been reported in several malignancies, including prostate cancer and acute myeloid leukemia. However, its function in renal cell carcinoma (RCC) remains unexplored. In this study, we investigated the role of CSRP1 in RCC for the first time.
CSRP1 and programmed death-ligand 1 (PD-L1) expression levels were determined using quantitative real-time polymerase chain reaction (qRT-PCR). The effects of CSRP1 overexpression on cellular proliferation, migration, and apoptosis were assessed in vitro through CCK-8, wound healing, and flow cytometry assays. To evaluate the role of CSRP1 in immunotherapy, Balb/c mice were treated with anti-PD-L1 antibody, and tumor growth was monitored.
In vitro, overexpression of CSRP1 significantly inhibited proliferation and migration of A498 cells while enhancing their sensitivity to sunitinib treatment. Mechanistically, CSRP1 overexpression downregulated PD-L1 expression in RCC cells. In BALB/c mice inoculated with Renca cells, CSRP1 overexpression led to reduced tumor growth and improved response to anti-PD-L1 therapy.
CSRP1 may play a role in regulating cell viability, migration, drug resistance, and possibly innate immunity in RCC. These findings suggest that CSRP1 could increase the efficacy of targeted drugs and immunotherapy in combination treatment strategies for RCC.
Hepatocellular carcinoma (HCC) is as the most frequently observed histological subtype among primary liver malignancies. While quercetin (QT) shows potential antitumor activity, its preclinical anti-HCC effects and safety (especially in animals) remain unclear. Most existing studies use single methods (e.g., individual animal or in vitro assays), which compromises the reliability of the conclusions. This study’s novelty lies in its use of a combined approach—integrating meta-analysis to quantify efficacy and network pharmacology to explore mechanisms, with experimental validation—to address this research gap. This work explores QT’s preclinical anti-HCC effects and adverse effects using this integrated approach.
We collected literature on the treatment of HCC with QT from January 2000 to August 2024. Nine articles meeting the requirements were included in the current study. Subsequent to this, a meta-analysis was conducted, with further validation via network pharmacology approaches and experimental assays.
A meta-analysis found that QT significantly inhibited HCC growth (reduced tumor volume/weight) and reduced mortality in tumor-bearing mice, with no significant effect on body weight. Network pharmacology identified protein kinase B alpha (AKT1) and the phosphoinositide 3-kinase (PI3K)/AKT pathway as potential therapeutic targets. Finally, the aforementioned conclusions were further verified through experimental validation.
Preclinically, QT effectively inhibited HCC growth and reduced mortality in tumor-bearing mice without affecting body weight, likely via the PI3K/AKT pathway (targeting AKT1). Our study results furnish preliminary evidence for QT as a promising candidate for HCC adjuvant treatment, supporting its further evaluation in clinical trials. Limitations include reliance on preclinical data; thus, the translational value needs clinical validation, and the underlying mechanisms require more in-depth investigation.
Although epirubicin is used among therapeutic options for multiple myeloma (MM), its clinical use remains limited, in part because the subgroup of patients most likely to benefit has not been clearly defined. Identifying robust biomarkers capable of predicting chemosensitivity is therefore essential to aimed personalized treatment strategies and enhance therapeutic outcomes. This study sought to characterize the molecular effects of epirubicin in MM cells, elucidate its tumor-suppressive mechanisms, and determine potential indicators for patient stratification.
The half-maximal inhibitory concentration (IC50) for epirubicin was quantified using the Cell Counting Kit-8 (CCK-8) viability assay. Gene expression alterations before and after epirubicin exposure were investigated via microarray profiling, followed by bioinformatic interrogation of publicly available datasets to examine the prognostic value of CDC20 expression in MM. Subsequently, functional validation was performed through in vitro assays and in vivo xenograft models to evaluate the impact of epirubicin on cell-cycle progression and tumor growth.
Epirubicin exhibited an IC50 of 23.85 μM in MM.1R cells. Transcriptome analysis revealed 115 genes upregulated and 25 genes downregulated post-treatment. Among the significantly altered genes were CDC20 (log FC = –2.409), KIF20A (log FC = –1.693), FAM72A (log FC = –1.742), CCNB1 (log FC = –1.787), PIF1 (log FC = –2.201), and LMNB1 (log FC = –1.589). Higher CDC20 expression was associated with shorter overall survival (OS), event-free survival (EFS), and post-progression survival (PPS). Mechanistic studies demonstrated that epirubicin triggers G2/M arrest in MM cells by suppressing CDC20, and in vivo experiments corroborated that decreased CDC20 expression contributes to reduced tumor proliferation via cell-cycle blockade.
Epirubicin exerts anti-myeloma effects by downregulating CDC20 and inducing cell-cycle arrest in MM, highlighting CDC20 as a potential biomarker for identifying MM patients likely to benefit from epirubicin.
In contemporary gene therapy research, recombinant adeno-associated virus (rAAV) has emerged as a pivotal delivery vehicle due to its favorable safety profile and capacity for sustained transgene expression. The most commonly utilized rAAV variants are hybrid vectors constructed by packaging the AAV2 genome within the capsid proteins of alternative serotypes (e.g., AAV1, AAV5, AAV8, or AAV9). This chimeric design combines the stable genomic integration and long-term expression characteristics inherent to AAV2 with the distinct tissue tropisms conferred by different capsid serotypes. Consequently, these engineered rAAVs exhibit enhanced organ-targeting specificity, enabling more efficient and selective gene delivery to desired tissues while minimizing off-target effects.
To identify the optimal hybrid vector for germ cell-directed gene delivery in mice, ten distinct chimeric AAV variants were generated by pseudotyping the AAV2 genome with diverse capsid proteins, followed by microinjection into seminiferous tubules via the efferent ducts. Transduction efficiency was comparatively evaluated at 4 weeks post-injection.
We demonstrated that AAV2/9 mediated robust and widespread enhanced green fluorescent protein (EGFP) expression in the mouse testis. Through immunofluorescence assays, we demonstrated that AAV2/9 demonstrated the capability to express in nearly all testicular cells, with remarkable efficiency and durability.
AAV2/9 demonstrated superior efficacy as a gene delivery vector for murine testicular germ cells without observable adverse effects on testicular development or spermatogenesis. This favorable safety profile, combined with high transduction efficiency establishes AAV2/9 as a promising candidate for therapeutic gene transfer to testicular germ cells. Collectively, our study identifies AAV2/9 as a premier vector for germ cell-directed gene therapy, providing crucial preclinical evidence to inform capsid selection for treating male infertility.
Temozolomide (TMZ) is a standard chemotherapeutic agent for glioma, but prolonged use frequently leads to drug resistance, reducing its therapeutic efficacy. Schisandrin B (Sch B), a lignan isolated from Schisandra chinensis, demonstrates promising anti-neoplastic activity. This study investigated the synergistic effects of Sch B and TMZ on U87 glioma cells to explore their combined influence on cell viability, apoptosis, and mitochondrial function.
U87 glioma cells were treated with Sch B, TMZ, or their combination. Cell viability was assessed using MTT assays. Apoptosis was evaluated by Hoechst staining and flow cytometry, while JC-1 staining and Western blotting were used to assess mitochondrial membrane potential, oxidative stress markers, and apoptosis-related proteins. Cell cycle analysis and pre-treatment with Z-VAD-FMK were performed to confirm pathway involvement.
Combination treatment significantly reduced cell viability (54.14%) compared to TMZ (72.47%) or Sch B (70.4%) alone. Flow cytometry indicated elevated apoptosis (22.3%) in the combination group. JC-1 staining and protein expression analyses revealed mitochondrial depolarization, cytochrome c release, activation of caspase-3 and -9, and a decreased Bcl-2/Bax ratio. The combined treatment induced G2/M cell cycle arrest via p53/p21 activation and increased oxidative stress. Pre-treatment with Z-VAD-FMK partially reversed these effects, confirming caspase-dependent mitochondrial apoptosis.
Sch B enhances TMZ-induced cytotoxicity in U87 glioma cells by promoting mitochondrial dysfunction, oxidative stress, and caspase-mediated apoptosis. These findings suggest that Sch B may serve as a promising adjuvant to improve the efficacy of TMZ-based glioma therapy, warranting further validation in resistant and in-vivo models.
Klebsiella pneumoniae is one of the most critical Gram-negative bacteria according to the World Health Organization (WHO). Due to the ability of this bacterium to evade antibiotics, phage therapy is becoming a promising tool. However, the use of isolated proteins rather than entire phages could reduce several risks associated with phage replication. Thus, understanding the protein composition and structural organization of bacteriophages is crucial for unlocking their biology and holds great potential for medicine and biotechnology.
In this study, artificial intelligence with AlphaFold 3.0 (AF3) and bioinformatic analysis were used to model the hitherto unknown structure of the Klebsiella phage KP32 (KP32), a complex and selective phage that targets K. pneumoniae strains with the K3 and K21/KL163 capsular serotypes.
By combining AF3 with sequence and structure analysis, we reconstructed the entire phage KP32. This complex phage is composed of over 500 protein chains, of which 415 compose its capsid and 104 its core-portal-tail complex, a platform that allows the phage to adhere to K. pneumoniae, hydrolyze its capsular sugars and finally inject its genetic code into the bacterium.
Phage therapy is a potentially promising tool for controlling antimicrobial resistance (AMR). However, one limitation arises from the limited knowledge of their nature and mechanisms of action, as only a few phages have been structurally characterized. The reconstruction of entire phages is currently a viable strategy for elucidating their mechanistic properties, knowledge that will enhance their potential applications as therapeutic alternatives.
Ultraviolet B (UVB) irradiation is a major environmental factor causing corneal epithelial cell apoptosis, leading to ocular surface damage and vision impairment.
This study aimed to investigate whether the standardized extract of Peucedanum japonicum Thunb. (SBP) protects corneal cells from UVB-induced apoptosis and explore its mitochondrial regulatory mechanisms.
Corneal epithelial cells were exposed to UVB irradiation, with or without treatment with SBP extract or its fractions. Nicotinamide adenine dinucleotide dehydrogenase activity, mitochondrial membrane potential, and mitochondrial morphology were assessed, and apoptosis-related proteins were analyzed using a cytokine antibody array kit. In vivo mouse models were also used to evaluate corneal damage following UVB exposure.
The SBP extract, particularly the n-butanol (n-BuOH) fraction, significantly attenuated UVB-induced mitochondrial dysfunction and reduced apoptosis. Treatment restored mitochondrial membrane potential and improved corneal morphology in UVB-exposed mice. Chlorogenic acid, a major active compound, exhibited similar protective effects. The n-BuOH fraction demonstrated protective effects comparable to those of chlorogenic acid.
SBP protects corneal cells from UVB-induced apoptosis through mitochondrial stabilization, suggesting its potential as a therapeutic agent for ocular surface disorders.
Hyperuricemic nephropathy is associated with mitochondrial dysfunction. Dynamin-related protein 1 (DRP1), a key regulator of mitochondrial fission, is activated under stress and translocates to the mitochondria, where it interacts with adapter proteins such as mitochondrial fission 1 protein (FIS1), thereby promoting excessive mitochondrial fission and apoptosis. Recent research has shown that inhibiting the DRP1/FIS1 interaction can reduce cellular injury in various disease models; however, its role in hyperuricemic nephropathy is unclear.
An in vitro model of hyperuricemic nephropathy was established by treating human renal tubular epithelial cells with uric acid (UA). Reverse transcription quantitative PCR and western blotting, and enzyme-linked immunosorbent assays were used to quantify the mRNA and protein levels of the target molecules. A specific peptide inhibitor, P110, was used to disrupt the binding between DRP1 and FIS1. Co-immunoprecipitation (Co-IP) was performed to confirm the interactions between DRP1 and FIS1. Cell viability was assessed using propidium iodide staining and the Cell Counting Kit-8 assay.
UA significantly upregulated DRP1 expression, activated DRP1, and promoted mitochondrial translocation. P110 inhibited DRP1/FIS1 binding, preventing DRP1 UA-induced mitochondrial translocation. Excessive mitochondrial fission, reactive oxygen species generation, release of inflammatory factors, and apoptosis were significantly alleviated. In addition, inhibition of DRP1 mitochondrial translocation decreased the expression of apoptosis-related markers and apoptosis.
The overactivation of DRP1 is crucial for UA-induced renal tubular epithelial cell injury. P110 exerts a cytoprotective effect by inhibiting the DRP1/FIS1 interaction and modulating the mitochondrial apoptotic pathway. This study proposes a possible target for therapeutic intervention in the treatment of hyperuricemic nephropathy.
Programmed death-ligand 1 (PD-L1) partners with specificity Protein 1 (SP1) or signal transducer and activator of transcription 3 (STAT3) to modulate the transcription of growth arrest-specific 6 (GAS6) and early growth response protein 1 (EGR1), necessitating mediators to avoid feedback. Based on binding and stemness data, high mobility group A1 (HMGA1) and Small Mother Against Decapentaplegic3 (SMAD3) were identified as potential mediators in this context. While the SMAD3–P300–STAT3 complex facilitates SMAD3–STAT3 crosstalk, it remains unclear whether the PD-L1–HMGA1–SP1 or PD-L1–SMAD3–SP1 complexes bind to GAS6 and EGR1 promoters to regulate their transcription.
MG63 osteosarcoma cells and SW620 colon cancer cells with unidentified nuclear PD-L1 function were chosen for our study. Chromatin immunoprecipitation and co-immunoprecipitation assays were performed to evaluate SP1, HMGA1, SMAD3, STAT3, P300 and PD-L1 (also denoted CD274) enrichment at the GAS6 and EGR1 promoters; the existence of the PD-L1–(HMGA1 or SMAD3)–SP1 complexes; whether P300 binds to STAT3; and whether HMGA1 and SMAD3 bind to P300. The alterations in GAS6, EGR1 and PD-L1 mRNA levels after their combined over-expression and/or knockdown were assessed via qPCR. Two representative target genes identified via PD-L1 chromatin immunoprecipitation (ChIP)-seq were examined to determine whether HMGA1 and SMAD3 were enriched at their promoters.
PD-L1, HMGA1, SMAD3, SP1, P300 and STAT3 were enriched at GAS6 and EGR1 promoters in two cell lines. HMGA1 or SMAD3 antibody pulled down PD-L1 and SP1; PD-L1 antibody pulled down HMGA1, SMAD3 and SP1; P300 antibody pulled down STAT3; and, surprisingly, HMGA1 and SMAD3 antibodies pulled down P300. Combined over-expression or knockdown significantly altered GAS6, EGR1 and PD-L1 mRNA levels. PD-L1 ChIP-seq indicated 114 target genes, among which PD-L1 and beta-transducin repeat containing E3 ubiquitin protein ligase (BTRC) were chosen to verify the promoter enrichment of HMGA1 and SMAD3.
Our study provides initial evidence that PD-L1 might form HMGA1- and SMAD3-dependent complexes to bind the GAS6, EGR1 and CD274 promoters, thus modulating the transcription of GAS6, EGR1 and PD-L1 mRNA in cancer and sarcoma cells.
Natural killer (NK) cells are crucial in inflammatory skin diseases, but their diverse functions and lack of a standardized classification system across diseases have limited deeper insights into their roles.
We merged single-cell transcriptomic data from 40 skin samples to create a comprehensive atlas of NK cells across various skin diseases, identifying nine distinct NK cell subsets with unique functions.
Our analysis revealed a conserved Aryl Hydrocarbon Receptor (AHR)+ NK cell subset that is broadly present across multiple skin diseases. Notably, the Granzyme B (GZMB)+ NK cell subset may be associated with the pathogenesis of psoriasis (PSO) and appears to undergo a differentiation trajectory toward IL13+ NK cells in the pseudotime analysis. This finding suggests a potential role for these cells in mediating paradoxical cutaneous inflammation. Our analysis identified NK cells expressing GZMB in a variety of skin diseases. Notably, the NK cell subpopulation expressing GZMB appears to be associated with the pathogenesis of PSO and ultimately exhibits the expression of IL13 in pseudotime analysis, suggesting that it may play a role in regulating contradictory skin inflammation.
This study offers a comprehensive overview of skin NK cells, identifies pathogenic subsets that may drive skin disease progression, and provides novel insights for future targeted therapies.
This study investigated necroptosis-related molecular alterations in the endometrium of patients with polycystic ovary syndrome (PCOS) using quantitative proteomic analysis and developed a predictive model for pregnancy outcomes based on these findings.
Liquid chromatography-tandem mass spectrometry was used to identify and quantify endometrial proteins. Differentially expressed proteins (DEPs) were screened and subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses to identify key pathways. Candidate prognostic necroptosis–related proteins were obtained by intersecting DEPs with the necroptosis gene set, followed by univariate Cox and Least Absolute Shrinkage and Selection Operator (LASSO) regression analyses to select those associated with pregnancy outcomes and construct a predictive model.
A total of 611 DEPs were identified (132 upregulated and 479 downregulated). KEGG enrichment revealed significant involvement of the necroptosis pathway. Six necroptosis-related proteins were identified using Cox and LASSO regression analyses and used to construct the predictive model. Kaplan–Meier analysis showed that the low-risk group had significantly better pregnancy outcomes than the high-risk group. The model achieved an area under the receiver operating characteristic curve of 0.903 for predicting live birth at 37 weeks, and decision curve analysis demonstrated superior clinical benefit compared to conventional clinical indicators. Furthermore, correlation analysis revealed significant associations between necroptosis-related proteins and classical endometrial receptivity markers, suggesting potential molecular crosstalk.
Proteomic profiling revealed enrichment of the necroptosis pathway in the endometrium of patients with PCOS. The constructed model indicated preliminary predictive potential for pregnancy outcomes, suggesting that necroptosis may contribute to impaired endometrial receptivity.