Colorectal cancer (CRC) remains the third most common malignancy and the second leading cause of cancer-related death worldwide, with limited biomarkers available for accurate prognostic stratification. This study integrates multi-cohort transcriptomic data from four GEO datasets (GSE39582, GSE17536, GSE14333, GSE33113; n = 1123) and the TCGA-CRC cohort (n = 621) to identify key differentially expressed genes and construct a validated prognostic risk score model. Differential expression analysis using the limma framework identified 642 upregulated and 588 downregulated genes (|log2FC|>1, FDR<0.05). Weighted gene co-expression network analysis (WGCNA) extracted hub genes from modules significantly correlated with overall survival. LASSO Cox regression then reduced the hub gene set to an eight-gene prognostic signature (MMP1, AURKA, TOP2A, CDH1, COL1A1, CLDN1, ETV4, CDX2) that stratified patients into high- and low-risk groups with significantly different survival outcomes (p < 0.001, log-rank test). The model achieved AUCs of 0.824, 0.796, and 0.768 for 1-, 3-, and 5-year overall survival in the training cohort and was validated in three independent datasets. High-risk patients exhibited immunosuppressive microenvironments (elevated M2 macrophages, Tregs) and increased resistance to standard chemotherapy agents. Integration of the risk score with clinical variables improved prognostic accuracy (C-index 0.812). These findings offer a robust bioinformatics-derived prognostic tool with implications for immunotherapy patient selection and personalized treatment planning in CRC.

Due to the poor prognosis of hepatocellular carcinoma (HCC), there is an urgent need to identify novel therapeutic targets. This study aims to evaluate the potential of Aurora kinase B (AURKB) as a therapeutic target for HCC and provide a theoretical basis for immunotherapy in HCC. In this study, 374 patients were included from The Cancer Genome Atlas (TCGA) database, and single-sample gene set enrichment analysis was performed to classify them into high-immunity and low-immunity groups. Through differential gene expression analysis and univariate Cox regression analysis, AURKB was screened out as a risk gene, followed by Kaplan-Meier survival analysis, immune infiltration analysis, and pathway enrichment analysis. The results demonstrated that overexpression of AURKB was associated with worse overall survival (OS) and disease-free survival (DFS) in HCC patients. High AURKB expression was correlated with increased infiltration of CD8 + T cells, macrophages, and dendritic cells, along with elevated expression of immune checkpoint molecules, including programmed death-1 (PD-1), cytotoxic T lymphocyte-associated antigen-4 (CTLA-4), CD80, and CD86. To further validate the role of AURKB in HCC, we constructed HCC cell lines with AURKB knockdown. Knockdown of AURKB inhibited HCC cell proliferation, colony formation, migration, and invasion. Functional experiments further indicate that AURKB promotes CD8 + T cell exhaustion and immune evasion. In conclusion, our study indicates that AURKB is an immune-related prognostic biomarker that promotes immune escape and tumor progression in HCC.

With the emergence of 3D bioprinting, the biofabrication of artificial skin has become possible through the precise arrangement of cells that can imitate the complexities of the skin tissue structure and functions. The current paper reviews concepts on skin drug delivery, paying special attention to the impact of skin physiology, barriers, and design of formulations on clinical performance. 3D printing technology refers to the diversity of technologies that transform computer-based designs into physical objects. Several of these techniques have been investigated for the fabrication of delivery systems and for surface functionalization through drug-loaded coatings. Biomedical applications of 3D bioprinting encompass burn management, accelerated wound healing, disease modeling, alopecia treatment through follicle regeneration, insulin delivery via microneedle systems and targeted anticancer therapies. With the incorporation of 3D bioprinting into the cosmetic and pharmaceutical sectors, it is now possible to manufacture customized skin care products. The review also examines the clinical significance and toxicological considerations associated with 3D-printed constructs, along with current regulatory frameworks governing their translation to clinical practice. Addressing these regulatory and translational barriers will be necessary to achieve clinical translation and commercialization of 3D-bioprinted skin constructs, accelerating next-generation personalized therapeutic and cosmetic applications.

Histochemical staining patterns may vary with age because of changes in cell number, differentiation state, marker expression, tissue geometry, or pre-analytical conditions. Recent data indicate that epithelial tuft-cell abundance decreases and LGR5 immunoreactivity changes in the ageing human colon. We emphasize that marker-positive structures, cell number, and biological function are related but non-equivalent outcomes, particularly for rare epithelial populations and the cellularly complex enteric nervous system. To strengthen interpretation and international reproducibility, we propose a minimum framework combining orthogonal cell-identity markers, transcript-level confirmation in representative samples, donor-level statistical inference, transparent reporting of fixation, staining, imaging, and segmentation variables, and anatomically resolved denominators. A tiered validation pathway could enable laboratories with different technical capacities to generate comparable evidence through standardized bright-field histochemistry, regional multiplex-imaging hubs, and targeted analysis of discordant specimens. This approach could help define age-related staining differences as continuous spatial phenotypes for mechanistic investigation and clinical ageing research.

In a recent Letter to the Editor, Dutt et al. have presented interesting arguments to improve the application of different therapies to melanoma. Although sharing most of their arguments, we feel it is necessary to clarify better some points in relation to the photothermal therapy of cancer applied to melanomas. In particular, when it has to do with continuous photothermal excitation. In this case, in our opinion, the need to assess in depth the features of the melanoma target relaxes, because heat will diffuse from the heating particles and affect the whole treated volume in a more or less homogeneous way.

Angiogenesis during endochondral ossification is crucial to skeletal development, bone regeneration, and fracture healing. Vascular buds at the chondro-osseous border represent key markers of angiogenic activity. Conventional quantification approaches, such as global vessel density measurements or manual counting of buds, are time-consuming and often lack reproducibility. Therefore, we developed SACOB (Semi-Automated Counting of Buds), a modular Python-based software pipeline designed for reproducible and anatomically guided quantification of vascular buds in histological sections of chondro-osseous tissues. The SACOB workflow integrates image import, user-defined path annotation along the cartilage-bone interface, extraction and optional smoothing of intensity profiles, and algorithmic peak detection to identify vascular buds. Validation was performed on Endomucin-stained femoral sections from C57BL/6 mice aged 4, 12, and 18 months (n = 18). Four sections per animal, derived from central and peripheral regions, were analyzed to assess both age-related and spatial differences in angiogenesis (n = 6 animals/group; 4 sections/animal (total sections per age group = 24), balanced by sex). The software reliably detected vascular buds and reproduced biological patterns of vascular decline with aging. Bud density was highest in 4-month-old mice (22.23 ± 3.06 mm[-1]), significantly reduced at 12 months (9.97 ± 1.54 mm[-1], p < 0.001), and further declined at 18 months (6.71 ± 2.11 mm[-1], p < 0.001). No significant differences were observed between central and peripheral femoral sections, indicating that angiogenesis activity is spatially consistent across the growth plate. Using SACOB workflow enables rapid, reproducible determination of vascular buds at the chondro-osseous border in tissue sections.

Stockert and Blázquez-Castro have skilfully changed the perspective on melanin, turning it from a histological pigment to a possible therapeutic substrate in melanoma. However, if melanin is considered a binary and/or spatially uniform property, then translation may be restricted. The amount of pigment, its molecular structure, redox status, and intracellular localization and interactions with living tumour and immune cells could account for the ability of melanin to shield from photodynamic damage, enhance photothermal damage, or represent a photodynamic damage-resistant state. We suggest a spatial melanin phenotype, based on pigment burden, physicochemical state, cellular compartment, microenvironmental context and optical response. A combination of validated histochemistry and image analysis (digital image) with selected spectroscopic or spatial method might be a resource adaptive workflow. The hypothesis of prospective studies is whether this phenotype predicts temperature distribution, and oxidative injury, immune activation and residual viable tumour more accurately than conventional pigmentation grading. This would adjust the place of histochemistry as a tool of phototherapy selection and dosimetry that can be used for action.

Cytotoxic brain edema is a life-threatening condition arising from pathological water accumulation within brain parenchymal cells, primarily astrocytes, following metabolic or toxic insults. Aquaporin-4 (AQP4), the predominant water channel in the central nervous system, is highly enriched in the perivascular endfeet of astrocytes and is considered a key facilitator of bidirectional water flux. Despite extensive molecular and light-microscopic studies, the precise nanoscale distribution of AQP4 within endfoot membranes and its spatial reorganization during the development of cytotoxic edema remain insufficiently characterized. We employed a high-resolution post-embedding immunogold electron microscopy protocol on brain tissue from an established murine model of water-intoxication-induced cytotoxic edema. Adult C57BL/6 J mice were subjected to acute systemic hypo-osmolality by intraperitoneal water loading, and perfused at defined time points. Ultrathin sections of frontoparietal cortex embedded in hydrophilic resin were labeled with primary anti-AQP4 antibodies and secondary antibodies conjugated to ultra-small (1.4 nm) or 10 nm gold particles, followed by silver enhancement for size-based discrimination of two gold particle populations. Quantitative analyses included gold particle density mapping along astrocytic plasma membrane domains, nearest-neighbour distance measurements, polarization indices, and double-label colocalization studies with the inwardly rectifying potassium channel Kir4.1. In control brains, AQP4 immunoreactivity was strikingly polarized, with a 12.7-fold higher gold particle density on perivascular endfoot membranes (62.4 ± 4.8 particles/µm) compared to the parenchymal-facing membranes of the same cells (4.9 ± 1.2 particles/µm; p < 0.001). Double immunogold labeling revealed that 73.5% of AQP4-positive clusters in endfeet colocalized with Kir4.1 signals. Following induction of cytotoxic edema, the density of AQP4 gold particles on endfoot membranes decreased by 41% within 15 min and by 64% at 60 min (p < 0.001), accompanied by a reciprocal increase in labeling on non-endfoot membranes. The polarization index collapsed from 12.7 to 2.1 at 60 min. Morphometric analysis documented a 3.2-fold increase in endfoot cross-sectional area and a progressive loss of orthogonal array-like particle clusters. Across pooled groups, AQP4 polarization was inversely associated with endfoot cross-sectional area; however, within-group analyses indicated that this association was largely attributable to treatment-group differences. This study provides a time-resolved quantitative ultrastructural analysis of AQP4 labeling across astrocytic membrane domains during the evolution of cytotoxic edema. These findings demonstrate a marked time-dependent loss of perivascular AQP4 polarization during acute cytotoxic edema and provide a quantitative framework for future studies examining the mechanisms underlying changes in AQP4 membrane organization.

Hepatitis B virus (HBV) infection remains a major global health challenge, requiring accurate diagnostic tools and precise therapeutic drug monitoring (TDM) to guide antiviral treatment and prevent disease progression. Conventional laboratory‑based assays, while reliable, are often limited by long turnaround times, high operational costs, and the need for specialized infrastructure. Recent advances in nanomaterials have transformed the landscape of HBV diagnostics and TDM by enabling highly sensitive, rapid, and cost‑effective analytical platforms. Nanostructures such as gold nanoparticles, quantum dots, carbon nanotubes, graphene derivatives, and magnetic nanomaterials offer unique physicochemical properties including large surface area, tunable optical signatures, and enhanced biorecognition capabilities that significantly improve assay performance. These materials have been integrated into diverse sensing modalities, including electrochemical biosensors, fluorescence‑based assays, plasmonic platforms, and point‑of‑care devices, achieving ultralow detection limits for HBV DNA, HBsAg, and other clinically relevant biomarkers. In therapeutic monitoring, nanomaterial‑enabled sensors facilitate real‑time quantification of antiviral drugs such as tenofovir and entecavir, supporting personalized treatment strategies and improving adherence assessment. Despite promising advancements, challenges remain regarding clinical translation, standardization, biocompatibility, and large‑scale manufacturing. This narrative review summarizes current nanomaterial‑based approaches for HBV detection and TDM, highlights emerging technologies, and discusses future directions for integrating nanodiagnostics into routine clinical practice to enhance HBV management and global disease control.

Hydroxyurea (HU) is a growth-inhibiting agent used to treat conditions characterized by excessive cellular proliferation. As an antimetabolite, HU inhibits ribonucleotide reductase, reducing the deoxyribonucleotide pool and thereby slowing DNA synthesis. Pharmacological inhibition of cell division is closely associated with the development of a cellular senescence phenotype, which may contribute to therapeutic resistance and tumor-associated inflammation. This phenotype has extensive effects, potentially inducing bystander effects and fostering tumor immunosuppression, thereby increasing the risk of cancer recurrence or relapse. Furthermore, HU has been shown to induce cellular senescence in non-malignant cells in laboratory settings. This review examines the principal molecular mechanisms underlying HU's cytotoxic and genotoxic effects, as well as the salient features of cellular senescence induction. A comprehensive understanding of the pathways through which HU prompts cellular senescence in normal cells may enhance current chemotherapy strategies and mitigate adverse effects in patients with cancer and other pathologies.

Tuberculosis (TB), caused by Mycobacterium tuberculosis (M.tb), remains a major global health threat. The tripartite motif (TRIM) family of E3 ligases plays crucial roles in various cellular processes, yet the involvement of TRIM32 in TB remains poorly understood. This study explores the function of TRIM32 in M.tb infection and its underlying mechanisms. We found that M.tb infection significantly reduced TRIM32 expression in mouse macrophages, suggesting a regulatory role in TB progression. TRIM32 inhibited NLRP3 inflammasome activation and pyroptosis of M.tb-infected macrophages. Furthermore, TRIM32 overexpression mitigated mitochondrial dysfunction, evidenced by decreased reactive oxygen species (ROS) generation, inhibited mitochondrial ROS (mtROS), and restored mitochondrial membrane potential. Importantly, TRIM32 directly interacted with TLR4 and promoted its degradation via ubiquitination at the K48 site, a critical process for proteasomal degradation. The inhibitory effects of TRIM32 on pyroptosis and mitochondrial dysfunction were partially reversed by TLR4 overexpression, confirming TLR4 as a key mediator of TRIM32's actions. Our findings uncover TRIM32 as a critical regulator of mitochondrial health and pyroptosis in M.tb-infected macrophages through TLR4-mediated ubiquitylation, Moreover, silencing of endogenous TRIM32 aggravated NLRP3 inflammasome activation, pyroptosis and mitochondrial injury, and AAV-mediated TRIM32 overexpression alleviated lung pathology, lowered bacterial burden and improved survival in M.tb-infected mice, with these benefits being partially abrogated by TLR4 co-overexpression. Our findings identify TRIM32 as a candidate regulatory node of macrophage pyroptosis in TB that merits further preclinical evaluation.

Ambystoma mexicanum has emerged as an important experimental model in regenerative biology due to its remarkable regenerative capacity and neotenic condition. Although histological descriptions of several organs have been previously reported, an integrative characterization of the tissues and organ systems of neotenic axolotls remains limited. To investigate the structural features and cellular organization of axolotl organs, we applied a systematic histological approach with comparative perspectives. Organ samples from neotenic axolotls were processed using conventional histological techniques and analyzed by light microscopy. Fundamental tissues and multiple organ systems, including digestive, cardiorespiratory, urinary, reproductive, tegumentary, nervous, endocrine, and lymphohematopoietic systems, were characterized. The analysis revealed several distinctive features, including cartilage characterized by predominantly isolated chondrocytes, trabeculated myocardial organization, epithelial transitions in the lung, the organization of exocrine pancreatic acini and endocrine islets, distinctive retinal organizational features, and extensive hematopoietic tissue associated with the liver capsule and pericardial serosa. In addition, this study provides detailed histomorphological descriptions of the pancreas and urinary bladder in neotenic axolotls. Overall, the findings demonstrate that while many tissues share organizational similarities with other amphibians and vertebrates, A. mexicanum exhibits particular structural adaptations potentially associated with neoteny and regenerative biology. This work expands the existing histological knowledge of the species and supports future studies in regeneration, developmental biology, comparative anatomy, pathology, and conservation.