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.
Background Decalcifying mineralized samples for microscopic analysis presents significant challenges. Aim To evaluate the effects of different decalcification solutions on Safranin O staining and immunohistochemistry for aggrecan. Materials and methods The paws of AG/WT, BALB/c, C57, DBA1/J mice, and Wistar rats were removed, fixed, and randomly divided into three groups: 10% nitric acid, 12.5% EDTA at room temperature, and 12.5% EDTA at 35°C with shaking. Tissue sections were prepared using a microtome and stained with Safranin O and an anti-aggrecan antibody. The staining intensity was then assessed. Results No statistically significant difference was found between the EDTA at room temperature and EDTA at 35°C for Safranin O staining. However, EDTA-treated samples generally exhibited stronger staining. For aggrecan quantification, the nitric acid protocol resulted in heterogeneous labeling among the studied strains. Notably, this protocol showed a statistically significant decrease in staining intensity in AG/WT and DBA1/J strains compared to the EDTA protocol. Conclusion The EDTA protocol produced more intense Safranin O staining compared to nitric acid across all strains. In terms of aggrecan detection, the AG/WT and DBA1/J strains exhibited reduced immunohistochemical reactivity when decalcified with nitric acid compared to EDTA.
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.
