The central nervous system (CNS) has a limited regenerative capacity, rendering traumatic injuries such as spinal cord injury (SCI) and traumatic brain injury (TBI) highly disabling and difficult to treat. These insults trigger complex pathophysiological cascades, including extensive cell death, sustained inflammation, and the formation of a hostile inhibitory microenvironment that compromises neural plasticity and hampers tissue regeneration. The multifactorial nature of these mechanisms, together with a fragmented understanding of CNS plasticity, has hindered the development of effective therapeutic interventions. In recent years, cell-based therapies have emerged as promising strategies to support neural repair and induce pro-regenerative processes. This approach encompasses multiple cell types, including bone marrow-derived mesenchymal stem cells (BMSCs), adipose-derived stem cells (ASCs), umbilical cord mesenchymal stem cells (UCMSCs), as well as neural progenitor cells (NPCs) and olfactory ensheathing cells (OECs). Clinical studies in SCI have reported functional improvements, particularly following treatment with BMSCs and peripheral blood-derived stem cells, although substantial methodological heterogeneity limits definitive conclusions. In TBI, clinical evidence remains more limited; however, preclinical studies consistently demonstrate the neuroprotective and regenerative potential of mesenchymal stem cell-based therapies. Beyond direct cell transplantation, increasing attention has been given to cell-free approaches, including secretome and extracellular vesicle-based strategies, which recapitulate many of the beneficial effects while potentially overcoming safety and logistical constraints. Overall, both cell-based and cell-free therapeutic strategies show significant potential to enhance neuroplasticity, attenuate secondary injury, and promote functional recovery following CNS trauma. Nevertheless, successful clinical translation will require larger, well-controlled trials and the establishment of standardized protocols addressing optimal timing, dosage, and routes of administration.
Cervical cancer harbors a profoundly immunosuppressive tumor microenvironment (TME) that impairs innate and adaptive antitumor immunity and, critically, limits the efficacy of emerging radioimmunotherapy strategies. The NKG2D receptor-ligand axis-comprising the stress-inducible ligands MICA and MICB-constitutes a pivotal innate immune recognition interface whose surface expression on tumor cells determines susceptibility to NKG2D-armed effector cells and, by extension, dictates the targetability of radiolabeled NKG2D-directed probes for precision radionuclide therapy (RNT). Yet the mechanistic basis for NKG2D ligand dysregulation and its implications for radionuclide theranostics in cervical cancer remain poorly defined. This study integrated single-cell RNA sequencing (scRNA-seq) and experimental validation to comprehensively map the NKG2D-axis immune escape landscape in cervical carcinogenesis and to delineate its translational significance for precision RNT target selection and patient stratification. scRNA-seq datasets (GSM1551311 and GSM1551411) were processed using Seurat and Harmony for cell-type annotation, immune landscape characterization, and radionuclide target density profiling. Louvain clustering was performed at a resolution of 0.8 after evaluating multiple resolution parameters (0.4-1.2) using the clustree package to ensure stable cluster assignments. The top 20 principal components were retained for Uniform Manifold Approximation and Projection (UMAP) embedding based on elbow plot analysis. Harmony integration used default parameters (θ = 2 and λ = 1) with convergence assessed over 20 iterations. Doublet detection was performed using DoubletFinder (v2.0.3) with an estimated doublet rate of 4.0%; additionally, cells with >40% ribosomal protein gene reads were excluded. Batch correction quality was validated using the Local Inverse Simpson's Index, Adjusted Rand Index, and silhouette coefficient metrics. Real-time quantitative PCR and enzyme-linked immunosorbent assay (ELISA) quantified expression of four candidate RNT-relevant genes-MICA, MICB (NKG2D ligands; primary radionuclide targeting molecules), SUSD1 (immunosuppressive upregulator; potential RNT resistance mediator), and STAG3L1-in HeLa, SiHa, and normal HCerEpiC cell lines. Five independent biological replicates were performed per cell line, each with three technical replicates, following Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines. Shapiro-Wilk normality testing and Levene's test for homogeneity of variance were applied prior to all parametric analyses. Cervical cancer scRNA-seq profiles revealed significantly depleted cluster of differentiation 8 (CD8)+ T cells (mean difference: -0.12; 95% CI: [-0.16, -0.08]; Cohen's d = 1.45) and natural killer (NK) cells (Cohen's d = 1.12), with increased CD25+ regulatory T cells (+0.08; 95% CI: [+0.05, +0.11]), establishing an RNT-unfavorable immunosuppressive TME. Comparative benchmarking against RNT-responsive tumor types, neuroendocrine tumors and prostate-specific membrane antigen (PSMA) positive prostate cancer, confirmed that cervical cancer exhibits a combination of reduced target surface density, depleted NKG2D-effector populations, and enriched immunosuppressive subsets collectively predictive of attenuated RNT efficacy. Experimental validation confirmed dramatic downregulation of MICA (HeLa: 0.44 ± 0.07 relative expression, p < 0.001, n = 5) and MICB (HeLa: 0.51 ± 0.09, p < 0.05), translating to markedly reduced MICA protein secretion (124.3 ± 18.5 pg/mL in HeLa versus 285.4 ± 31.2 pg/mL in controls, p < 0.01). Concurrently, SUSD1 was markedly upregulated (HeLa: 2.28 ± 0.25-fold; protein 3.42 ± 0.45 ng/mg, p < 0.001, n = 5). Strong mRNA-protein correlations, r = 0.78-0.92, p < 0.001; computed from five independent biological replicates per cell line; coefficient of variation (CV) < 15% for all measurements, validated transcriptomic profiling as a reliable proxy for theranostic target protein density estimation. This integrative study reveals that MICA/MICB downregulation and SUSD1 upregulation converge to suppress NKG2D-mediated antitumor immunity in cervical cancer, creating an immune-cold TME that limits current immunotherapy and radionuclide targeting efficacy. The NKG2D ligand expression landscape mapped here delineates a precision RNT strategy: scRNA-seq-guided patient stratification, radiolabeled anti-MICA/MICB nanobody theranostic imaging to confirm surface target density, and combination radioimmunotherapy integrating MICA/MICB re-expression induction with targeted radionuclide delivery to selectively irradiate the NKG2D-ligand-negative tumor cell population.
Multiple interacting factors within the tumor microenvironment, including mechanical pressure, extracellular matrix components, hypoxia, and vascular architecture, are known to promote cancer progression. Although fibronectin 1 functions as a critical extracellular matrix glycoprotein in colorectal cancer, its specific interaction with mechanical pressure is not well characterized. This study utilized a meta-analysis (PROSPERO: CRD42024571414) alongside an in vitro weight-induced compression model to evaluate the prognostic role of fibronectin 1 and its interaction with mechanical pressure during colorectal cancer progression. The meta-analysis demonstrated significantly elevated fibronectin 1 expression in colorectal cancer patients compared to controls (SMD = 0.90, 95% CI: 0.56-1.25, P < 0.001) and a positive association with distant metastasis (OR = 3.63, 95% CI: 1.21-10.95, P = 0.0219). Analysis of colorectal cancer tissues versus adjacent normal tissues (n = 19) revealed markedly increased fibronectin 1 expression in both tumor cells and stromal components. Single-cell RNA sequencing analysis (GSE302903) identified FN1 expression in diverse cell populations, including cancer-associated fibroblasts, endothelial cells, macrophages, and tumor cells. Furthermore, high fibronectin 1 levels were established as a poor prognostic factor in colorectal cancer (HR = 2.47, 95% CI: 1.88-3.25, P < 0.001). In vitro experiments showed that pressure enhanced viability, proliferation, migration, and invasion of colorectal cancer cells and cancer stem cells through fibronectin 1. RNA sequencing indicated significant pressure-induced gene expression changes in colorectal cancer cells and identified the activation of multiple pathways. Specifically, the combined effect of pressure and fibronectin 1 upregulated of TGFBR1, SMAD2, SMAD4, and ROCK1 gene and p-SMAD2 expression. The expression of fibronectin 1 predicts poor overall survival in colorectal cancer and, together with mechanical pressure, facilitates colorectal cancer progression.
To explore the promotion of dynamic distraction on osteogenic differentiation of human bone marrow mesenchymal stem cells (hBMMSCs) in three-dimensional culture. Dynamic stretching in a three-dimensional culture for hBMMSCs was achieved with proportions set at 5%, 10%, and 20%, a frequency of 0.5 Hz, and a dynamic stretching duration of 2 hours per day. Static culture was used as the control group. Deformation of hBMMSCs induced by dynamic stretching was observed through cytoplasmic fluorescence staining. After 7 days of dynamic stretching culture, the impact of dynamic stretching on the viability of hBMMSCs was observed through cell live/dead staining. The effect of dynamic stretching on the osteogenic differentiation of hBMMSCs was observed through alkaline phosphatase (ALP) staining and the expression of osteogenic related genes and proteins after 7 days of dynamic stretching culture. Dynamic stretching in a three-dimensional culture for hBMMSCs was successfully constructed, which could achieve different durations, frequencies, and ratios of dynamic stretching. Dynamic stretching led to deformation of hBMMSCs, and the greater the stretching ratio, the more pronounced the cell deformation, transitioning from a circular to a flat oval shape. After 7 days of dynamic stretching culture, hBMMSCs in the static control group and dynamic stretching groups were mostly green stained live cells, with only a few red stained dead cells. The difference in the proportion of live cells between the groups was not statistically significant (P>0.05). The ALP staining in the dynamic stretching group was deeper than that in the static control group, and the number of ALP staining positive cells observed under the microscope was higher. The expression of osteogenic related genes and proteins increased after 7 days of dynamic stretching culture, and the difference was statistically significant (P < 0.05). Among them, the dynamic stretching group with 10% had the deepest ALP staining, the highest number of positive cells, and the most significant increase in the expression of osteogenic related genes and proteins compared with the static control group. Dynamic stretching caused deformation of hBMMSCs without a significant impact on cell viability, and it could effectively promote the osteogenic differentiation of hBMMSCs.
Shugoshin proteins SGO1 and SGO2 regulate chromosome cohesion and segregation during mitosis and meiosis, and their aberrant expression is implicated in several cancers. However, the mechanistic involvement of these two proteins in papillary renal cell carcinoma (KIRP) and their contribution to immune evasion remains unclear. Therefore, in the present study, we aimed to determine SGO1 and SGO2 expression patterns and prognostic significance, clarify their functional involvement in driving tumor progression, and evaluate their impact on the tumor immune microenvironment. We performed an integrative analysis using multi-omics datasets retrieved from TCGA, GTEx, and GEO, complemented by single-cell transcriptomics, immunohistochemistry of patient samples, and functional assays in renal cancer cell lines. We also investigated the expression, clinical relevance, and mechanistic roles of SGO1 and SGO2 in KIRP progression and tumor immune regulation. SGO1 and SGO2 were significantly upregulated in KIRP, correlating with advanced tumor stage and poor overall survival. Receiver operating characteristic analyses demonstrated strong diagnostic performance, particularly when SGO1 and SGO2 were evaluated jointly. Functional knockdown experiments revealed that both SGO1 and SGO2 promote the proliferation, migration, and invasion of renal cancer cells. Gene enrichment analyses further linked these proteins to E2F signaling, genomic stability, and immune-related pathways. Importantly, high SGO1 and SGO2 expression was strongly associated with increased infiltration of immunosuppressive cells, including regulatory T cells, M2 macrophages, cancer-associated fibroblasts, and myeloid-derived suppressor cells, suggesting their potential contribution to an immune-evasive tumor microenvironment. SGO1 and SGO2 are associated with aggressive tumor biology in papillary renal cell carcinoma and correlate with remodeling of the tumor immune microenvironment toward an immunosuppressive state. Importantly, while these findings highlight a strong association with the tumor immune microenvironment, they remain correlative, and further mechanistic studies are warranted to establish a direct causal relationship. Nevertheless, their associations with both tumor progression and immunosuppressive features suggest potential as prognostic biomarkers and targets for future mechanistic studies.
Next-generation sequencing (NGS) technology was used to analyze the gene mutation profile of lymph node metastases in renal cell carcinoma, and the molecular characteristics associated with poor prognosis were found, providing new ideas for mechanism research and treatment. Retrospective clinical data collection was conducted on 31 patients with lymphoid metastatic renal cell carcinoma and 21 patients with non-metastatic renal cell carcinoma. A total of 81 formalin-fixed paraffin-embedded tissue samples were retrieved from the Department of Pathology, including primary tumor, lymph node metastasis, and distant metastasis samples. The gene mutation profiles of the patients were examined using next-generation sequencing technology. The patients were followed up to analyze the correlation between lymph node metastasis and patient prognosis. The lymph node metastasis group showed differences in tumor size (P=0.006), World Health Organization (WHO)/International Society of Urological Pathology (ISUP) grade (P=0.002), T stage (P=0.003) and tumor thrombus (P=0.025) compared with non-metastatic renal cell carcinoma. The most commonly mutated genes in our cohort were the tumor suppressor genes VHL (38%), PBRM1 (22%), and SETD2 (20%). More-over, copy number variations were associated with tumor metastasis, and some mutation features were highly similar to known mutation patterns. There was a difference in mutation frequency between the patients in the metastasis group and samples in the non-metastasis group. The mutation frequency of most genes in the metastasis group was higher, however, Reactome pathway enrichment analysis did not show statistically significant differences in the shared enriched pathways between the two groups. There was a strong degree of concordance between the tumor' s primary and metastatic foci in the same patient, and genomic indicators [such as purity, ploidy, weighted-genomic integrity index (WGII), and intra-tumor heterogeneity (ITH)] as well as clonal and subclonal composition analysis further supported this consistency. The overall survival (OS) was higher in the patients without metastases (P=0.041), and specific gene mutations (such as IGF2R, JUN, EPHA5, and FH) were associated with poorer prognosis. To facilitate distant metastasis, lymph nodes might function as a "metastatic pool". The multigene NGS evaluates multiple relevant markers simultaneously, revealing several genetic alterations in the patients with lymphatic metastatic renal cell carcinorma. NGS-based molecular analysis can assist clinicians in assessing a patient' s prognosis and identifying novel, potentially therapeutic mechanisms.
Infertility is a major global health challenge, and non-obstructive azoospermia (NOA) represents the most severe form of male infertility. While a small proportion of NOA cases are caused by congenital factors, the majority arise from unknown causes and are therefore classified as idiopathic non-obstructive azoospermia (iNOA). Understanding the pathogenesis of iNOA is essential for developing effective diagnostic and therapeutic strategies. In this study, we analyzed single-cell RNA sequencing (scRNA-seq) data from testicular tissues of patients with iNOA and healthy controls, and validated the findings using hematoxylin and eosin (H&E) staining, Masson's trichrome (MT) staining, and Multiplex Immunofluorescence of patient testicular sections. Our results identified peritubular myoid cells (PTMs) as the primary contributors to testicular interstitial fibrosis. Cell-cell communication analysis revealed that macrophage-derived PDGF signaling is significantly upregulated in iNOA and promotes PTMs fibrosis. Fibrotic PTMs were associated with changes in the immune microenvironment, including pro-inflammatory activation of macrophages, T cells, and mast cells, consistent with a potential fibrosis-inflammation interaction. Furthermore, through integrative network pharmacology and molecular docking analyses, we identified regorafenib as a potential therapeutic agent that targets PTM fibrosis. These findings offer new insights into the diagnosis and treatment of iNOA.
Colorectal carcinoma (CRC) ranks among the top five most common malignancies globally, characterized by a high incidence and poor prognosis. Hepatocyte growth factor receptor (HGFR), a tyrosine kinase oncoprotein, plays a critical role in tumor progression. This study aimed to investigate the effects of stylosin (STL), a natural monoterpene, on human CRC cells, with focus on its potential interactions with HGFR. For computational analysis, the expression of HGFR was assessed in CRC tissues, pharmacokinetics of STL were predicted, and the interaction between STL and HGFR was determined by molecular docking. For experimental studies, STL was isolated from Ferula ovina roots via thin layer chromatography, and its structure was confirmed by 1H NMR spectroscopy. LoVo cells were treated with STL at concentrations of 25, 50, and 100 μM for 24, 48, 72, 96, and 120 h. Cell viability was assessed by alamarBlue, while apoptosis was evaluated by annexin V-FITC/PI staining followed by flow cytometry analysis. Results demonstrated significant overexpression of HGFR in CRC tissues relative to normal samples. The pharmacokinetic analysis of STL predicted high bioavailability, favorable distribution, and low toxicity risks. Molecular docking predicted a favorable binding affinity of STL to the active site of HGFR. Treatment with STL led to a dose-dependent decrease in cell viability, with the most pronounced cytotoxic effect observed at 100 μM. This was supported by notable morphological changes and increased apoptosis in treated cells. Collectively, the present findings show anticancer activity of STL in CRC cells, with computational predictions suggesting possible HGFR involvement and favorable pharmacokinetic properties. These results provide initial evidence supporting further investigation and position STL as a potent agent against CRC.
Foundation models such as scGPT have demonstrated strong potential for single-cell multi-omics integration; however, their downstream performance is highly sensitive to hyperparameter selection. Manual fine-tuning remains computationally expensive, dataset-dependent, and often irreproducible. Despite the increasing adoption of foundation models in single-cell analysis, systematic strategies for robust hyperparameter optimization remain underexplored. We developed a Bayesian optimization framework based on Tree-structured Parzen Estimators (TPE) for automated fine-tuning of scGPT and evaluated its performance on two benchmark bone marrow mononuclear cell (BMMC) multi-omics datasets, including CITE-seq and GSE194122 datasets. Across datasets, Bayesian optimization consistently improved biological conservation and batch integration metrics compared with default scGPT configurations. On the original BMMC benchmark, optimization improved AvgBIO from 0.59 to 0.67 and PCR from 0.33 to 0.52. On the GSE194122 dataset, the default configuration exhibited unstable convergence and weak biological preservation (AvgBIO = 0.19; ARI = 0.007), whereas Bayesian optimization substantially improved integration performance (AvgBIO = 0.60; ARI = 0.63) while reducing validation loss from 137 to 47.1. These findings demonstrate substantial dataset-specific sensitivity of scGPT fine-tuning and highlight the importance of automated optimization for stable deployment across heterogeneous multi-omics datasets. Our study demonstrates that Bayesian optimization provides an effective and reproducible strategy for stabilizing scGPT fine-tuning across diverse single-cell multi-omics datasets. Rather than introducing a new integration architecture, this work emphasizes the importance of systematic optimization for improving robustness and reproducibility of foundation-model applications in computational biology. Our model and dataset are freely available at: https://github.com/daren642/scGPT_multiomic_tuning.
Tracheal defect repair is a major clinical challenge in thoracic surgery, with long-segment defects having a low clinical repair rate. Autologous cartilage transplantation is limited by high postoperative stenosis and insufficient autologous tissue, while traditional artificial tracheal materials have poor biocompatibility and no epithelial regeneration-inducing capacity, leading to poor long-term efficacy. This study aimed to fabricate a biomimetic composite hydrogel and evaluate its physicochemical properties and tracheal repair efficacy, providing a new tissue-engineered strategy for clinical tracheal defect management. A PEGDA/DCM/EGF composite hydrogel was prepared via photopolymerization. Its physicochemical properties and EGF release pattern were characterized. In vitro experiments on rat tracheal epithelial cells assessed cell viability, proliferation and migration. A tracheal defect model was established in SD rats, with the hydrogel implanted; gross observation, HE staining and immunohistochemistry were performed at 2 weeks post-implantation to evaluate repair efficacy, with statistical methods used for intergroup comparisons. The PEGDA/DCM hydrogel had optimized microstructure and mechanical properties matching native trachea, maintained structural stability under physiological conditions, and achieved sustained EGF release without burst effect. The PEGDA/DCM/EGF hydrogel significantly promoted the viability, proliferation and migration of rat tracheal epithelial cells in vitro. In vivo, implanted SD rats had intact tracheal architecture and unobstructed lumens at 2 weeks; HE staining and immunohistochemistry confirmed continuous epithelial layer formation and successful epithelialization at the defect site. The PEGDA/DCM/EGF composite hydrogel has favorable physicochemical properties, excellent biocompatibility and effective tracheal repair efficacy, with DCM optimizing hydrogel performance and sustained EGF release accelerating epithelial regeneration. This study provides a promising tissue-engineered strategy for tracheal defect repair, with significant clinical translation potential for thoracic surgery clinical practice.
Cancer remains a major global health challenge, with rising incidence and persistent therapy resistance highlighting the need for new multi-targeted therapeutic agents. Pinocembrin (5,7-dihydroxyflavanone), a naturally occurring flavanone found in honey, propolis, and several medicinal plants, has gained increasing attention for its broad-spectrum anticancer potential. Recent studies demonstrate that pinocembrin modulates multiple hallmarks of cancer by regulating the PI3K/Akt/mTOR, STAT3, and NF-κB pathways, resulting in antiproliferative, pro-apoptotic, anti-metastatic, and anti-inflammatory effects. Evidence from diverse cancer models-including breast, prostate, colon, lung, ovarian, and melanoma-shows that pinocembrin induces cell-cycle arrest, activates intrinsic and extrinsic apoptotic pathways, inhibits angiogenesis, and suppresses epithelial-mesenchymal transition. Structure-activity relationship (SAR) analyses further reveal that modifications such as hydroxylation, esterification, and glycosylation enhance its bioavailability and anticancer activity. Despite its therapeutic promise, the clinical utility of pinocembrin is limited by poor solubility and rapid metabolic clearance. Recent nanotechnology-based formulations, including polymeric micelles, liposomes, nanoparticles, and nanoemulsions, have significantly improved their stability, bioavailability, and tumor-targeted delivery. Pinocembrin also exhibits synergistic effects with conventional chemotherapeutics while maintaining low toxicity toward normal cells, underscoring its suitability for combination therapy and chemoprevention. Preliminary clinical data indicate a favorable safety profile, although long-term toxicity, optimal dosing, and pharmacokinetic parameters require further investigation. This review synthesizes current knowledge on the anticancer mechanisms, SAR-driven insights, nanotechnology-enhanced delivery, synergistic actions, and safety considerations of pinocembrin. By integrating recent findings and highlighting research gaps, it provides a comprehensive foundation for advancing pinocembrin toward future preclinical and clinical applications in oncology.
The absence of induced pluripotent stem cell (iPSC) lines derived from Emirati patients with developmental disease hampers region-specific disease modeling and therapeutic research. Herein, we describe the creation of an iPSC line from peripheral blood mononuclear cells obtained from a 21-year-old Emirati female patient with ventricular septal defect (VSD) using Sendai virus-mediated delivery of reprogramming factors. The resulting line, UAEUi001-A, exhibited typical colony morphology, was mycoplasma negative, successfully generated embryoid bodies), and demonstrated strong alkaline phosphatase activity. These iPSCs were further characterized for pluripotency potential and their differentiation potential into the three germ layers under in vitro culture conditions through immunostaining using stage-specific markers. To the best our knowledge, this is the first reported generation of an iPSC line from an Emirati patient with VSD. Overall, this iPSC line may serve as a valuable model for establishing an Emirati-specific iPSC repository, supporting disease modeling and drug discovery relevant to the Emirati population.Impact StatementThis study establishes the first induced pluripotent stem cell (iPSC) line derived from an Emirati patient with ventricular septal defect, addressing a critical gap in region-specific disease models. It provides a valuable platform for understanding developmental cardiac disorders in underrepresented populations and supports the development of precision medicine and targeted therapeutic strategies relevant to the Emirati population.
Hepatocellular carcinoma is a leading cause of cancer-related mortality. Several microRNAs play key roles in HCC development and progression. Epigenetic processes, such as DNA methylation, might regulate these RNAs during HCC pathogenesis. In this study, we show that the miR-379/656 cluster/C14MC acts as a tumor suppressor cluster and is epigenetically regulated by DNA methylation. C14MC miRNA expression was determined in HCC cell lines using the nCounter assay and from the TCGA-LIHC clinical cohort. C14MC putative promoter was identified, characterized using cloning and luciferase assay, and the methylation status of promoter-bound CpGs was determined using bisulfite Sanger sequencing. The expressions of C14MC targets were experimentally validated by transcriptomic sequencing or transfecting mimics, followed by qRT-PCR. Furthermore, the diagnostic and prognostic significance of C14MC and its target interactome in HCC was assessed using clinical data from the TCGA-LIHC cohort. C14MC was downregulated in HCC cell lines and in TCGA-LIHC. The loss of C14MC tumor suppressor function was directly regulated by the hypermethylation of promoter-CpGs. Reactivating specific C14MC miRNAs, such as miR-299-5p and miR-376c-3p via mimics, abrogated the expression of several target oncogenes, including PARP1, SPP1, RAD21, and CENPA, which regulate critical molecular pathways such as the p53 signaling and NF-κB signaling pathways in HCC. Additionally, overexpressing these miRNAs inhibited HCC cell migration and invasion. Also, C14MC and its target interactome exhibited significant clinical correlation in terms of survival outcomes of HCC patients. This is the first study to show that C14MC is a methylation-dependent cluster in HCC. Several of these miRNAs and their targets can be used for early HCC diagnosis and prognosis. Thus, targeting C14MC can be useful in HCC management.
Head and neck squamous cell carcinoma is a heterogeneous disease with poor survival outcomes. Bcl-xL regulates tumor cell survival and apoptosis and has been associated with metastasis and poor prognosis, although its overall role remains undeciphered. We systematically evaluated the prognostic value of Bcl-xL in head and neck squamous cell carcinoma following PRISMA 2020 guidelines. We identified 2241 reports from seven databases, of which 20 original studies with overall good quality (REMARK) and low bias (QUIPS) were included. An association between Bcl-xL levels and survival was rarely observed. However, high Bcl-xL levels correlated with increased risk of lymph node metastasis in 40% of studies. A study on oral tongue cancer identified a significant correlation between high Bcl-xL levels and decreased survival and lymph node metastasis. Current evidence does not support Bcl-xL as a general prognostic marker in head and neck squamous cell carcinoma. Nevertheless, Bcl-xL may have prognostic relevance in oral tongue cancer if further studies confirm the original finding.
ObjectiveCytomegalovirus reactivation is a clinically significant complication after allogeneic hematopoietic stem cell transplantation. Valganciclovir is commonly used for cytomegalovirus prophylaxis; however, the optimal dosing strategy remains unclear. This study compared the efficacy and safety of ultra-low-dose valganciclovir (450 mg every other day) and low-dose valganciclovir (450 mg daily) in preventing cytomegalovirus reactivation after allogeneic hematopoietic stem cell transplantation.MethodsWe retrospectively reviewed adult patients who underwent allogeneic hematopoietic stem cell transplantation from January 2020 to December 2023. Individuals receiving low-dose (n = 31) or ultra-low-dose (n = 35) valganciclovir as cytomegalovirus prophylaxis were included. Cytomegalovirus DNAemia was evaluated within the first 14 weeks and then at 14-28 weeks after transplantation. Cytomegalovirus disease, acute graft-versus-host disease, survival outcomes, and hematologic toxicities were also assessed.ResultsThe prevalence of early cytomegalovirus reactivation was 13.8% in the low-dose group and 20.0% in the ultra-low-dose group (p = 0.301). Late reactivation rates were similarly low in the low-dose and ultra-low-dose groups (3.6% vs. 6.2%, p = 1.000). Hematologic adverse events were comparable; however, treatment discontinuation due to cytopenia was significantly more common with low-dose therapy (48.4% vs. 17.1%, p = 0.007). Overall survival (80.6% vs. 75.8%, p = 0.839) and acute graft-versus-host disease (29.1% vs. 45.7%, p = 0.381) rates did not exhibit statistically significant differences between the two groups.ConclusionsUltra-low-dose valganciclovir demonstrated better tolerability but may provide less protection against cytomegalovirus reactivation.
While the central dogma outlines DNA-to-protein information flow, existing gene regulators mainly target transcription. Here, we developed the λN-Guided RNA Targeting System (λGRTS), a CRISPR-independent platform enhancing mammalian mRNA translation via specific translation-guiding RNAs (tgRNAs). λGRTS integrates λN (high-affinity BoxB binder), mutated eIF4E1 (ablated non-specific 5' cap binding, retains TIC recruitment), and auxiliary factors (HuR for dsRNA stabilization, PABP and RRM2-RRM3 for mRNA closed loops). Optimized 22-nt tgRNAs (targeting 58 bp upstream of mRNA ATG in 5' UTR) and NES-tagged proteins maximized efficacy. λGRTS outperformed dCasRX (smaller ~ 50 kDa vs. ~ 150 kDa, single tgRNA vs. crRNA-tracrRNA), boosting functional proteins (e.g., GFP). It activated P53/PTEN, suppressing GBM cell proliferation, inducing G1 arrest, and reducing invasion in vitro. In vivo, lentiviral λGRTS inhibited orthotopic GBM in nude mice, extended survival by approximately 35-40 days, with IHC confirming P53/PTEN upregulation. Mass spectrometry showed no off-target effects. Cross-species tests (human, mouse, bovine cells) validated broad applicability via conserved eIF4E1. λGRTS offers a specific, safe translation-centric tool for gene regulation and oncology research.
Stroke induces profound neuroinflammation in which macrophages play a complex dual role, contributing to both injury and repair. The traditional M1/M2 classification is increasingly recognized as oversimplified. Advances in single-cell RNA sequencing (scRNA-seq) have revealed a spectrum of dynamic macrophage subpopulations with distinct functional and metabolic states, fundamentally reshaping our understanding of post-stroke immunity. This review synthesizes recent insights into macrophage heterogeneity from a single-cell perspective, highlighting novel subsets such as an LCP1⁺ population defined by coupled glycolipid metabolism. We discuss how metabolic reprogramming, including glycolysis, oxidative phosphorylation, cholesterol metabolism, hypoxia‑driven gradients, and mitochondrial dynamics, critically underpins macrophage polarization. Glycolysis fuels pro-inflammatory (M1-like) responses, whereas oxidative phosphorylation and fatty acid oxidation support anti-inflammatory and reparative (M2-like) functions. We further explore innovative nano‑therapeutic strategies, including engineered liposomes, exosomes, and responsive polymeric nanoparticles, that enable spatiotemporally precise modulation of macrophage activity. Based on these advances, we propose an integrative framework that directly links scRNA‑seq‑defined macrophage subsets to their metabolic pathways, druggable targets, and tailored nano‑interventions. We also critically examine clinical translation barriers and prioritize actionable targets (e.g., CCR2, PPARγ, Nrf2) for future stroke therapy. The convergence of single‑cell genomics, immunometabolism, and nanotechnology offers a transformative path toward precision immunomodulation in stroke. Moving beyond the static M1/M2 dichotomy to target macrophage subpopulations and their metabolic drivers guided by an integrated framework holds significant promise for developing more effective therapies.
Pyruvate carboxylase (PC) replenishes tricarboxylic acid cycle intermediates, driving cancer metabolic reprogramming. To improve the metabolic stability of erianin, a potent PC inhibitor from Dendrobium chrysotoxum Lindl, we designed and synthesized 55 derivatives, culminating in the identification of CIB-Q22, which exhibited potent PC inhibition (IC50 = 1.74 nM) and suppressed HCC cell viability (IC50 = 25.18 nM), comparable to erianin. Notably, CIB-Q22 demonstrated significantly improved in vivo stability, with a half-life (T1/2 = 1.21 h) much longer than erianin (T1/2 ∼ 0.1 h). Mechanistically, CIB-Q22 suppressed HCC proliferation and metastasis by inducing apoptosis and ferroptosis. Moreover, it promoted mitochondrial oxidative stress and inhibited glycolysis, thereby sensitizing cells to glutamine deprivation. In vivo, CIB-Q22 exhibited comparable antitumor efficacy but improved safety compared to sorafenib. With its potent PC inhibition and favorable drug-like properties, CIB-Q22 represented a promising therapeutic candidate for HCC treatment.
This study aimed to recapitulate adipose-tumor interactions within the colorectal cancer (CRC) tumor microenvironment (TME) and to elucidate the role of adipose-derived stem cells (ADSCs) in regulating CRC progression through cytokine-mediated signaling. Human ADSCs were isolated from adipose tissue and directly cocultured with CRC cells using an oxygen-permeable, PDMS-based 3D coculture chip, followed by cytokine profiling of conditioned media, immunofluorescence analysis of spheroids, FACS-based cell separation, and molecular analyzes including western blotting, qPCR, and immunoprecipitation to interrogate HIF-1-related mechanisms. The results revealed that cancer-associated ADSCs secrete CCL8, which markedly enhances CRC cell migration. Mechanistically, ADSC-derived CCL8 activated the ERK signaling pathway in CRC cells, leading to increased HIF-1α protein accumulation without significant changes in protein stability. This was accompanied by enhanced interaction between HIF-1α and the transcriptional cofactor p300. Consequently, HIF-1α transcriptional activity was increased, resulting in the upregulation of downstream epithelial-mesenchymal transition markers and promoting a pro-migratory and aggressive cancer phenotype. These effects were particularly pronounced in the coculture system, where intensified crosstalk between ADSCs and cancer cells amplifies oncogenic signaling within the TME. Collectively, these findings demonstrate that ADSC-derived CCL8 functions as a key mediator of adipose-tumor crosstalk in CRC progression by driving HIF-1α-dependent signaling pathways. Furthermore, the PDMS-based 3D coculture platform employed in this study provides a robust and physiologically relevant experimental system for dissecting complex cell-cell interactions and cytokine-driven mechanisms within the TME of CRC.
Metabolic dysfunction-associated steatotic liver disease (MASLD) and its progressive inflammatory form, metabolic dysfunction-associated steatohepatitis (MASH), constitute major causes of hepatic morbidity worldwide. Central to the pathophysiology of these conditions is the progressive impairment of liver sinusoidal endothelial cells (LSECs), whose characteristic fenestrated architecture is indispensable for hepatic homeostasis. Capillarization, defined as the loss of LSEC fenestrations, propels disease progression by disrupting the equilibrium between pro-fenestration and pro-capillarization signaling cascades. This review critically appraises the LSEC Fenestration Signaling Nexus as a central, targetable mechanism in metabolic liver disease and assesses emerging therapeutic strategies engineered to restore fenestrations. We present a translational roadmap that prioritizes combinatorial pharmacological approaches and LSEC-specific biomarkers for precision medicine. This integrative framework furnishes new perspectives for rational drug development and individualized therapeutic strategies in metabolic liver disease.