Tamoxifen (TAM), a selective estrogen receptor modulator, competes with estrogen for binding to estrogen receptor (ER) sites, thereby inhibiting estrogen-induced proliferation in ER-positive breast cancer cells. However, long-term treatment with TAM often leads to resistance, and its mechanisms, particularly involving biochemical markers, remain unclear. This study investigated the effects of TAM (10-6 M and 10-8 M) compared to 17β-estradiol (10-6 M and 10-8 M) on galectin-3 and laminin expressions in ER-positive MCF-7 breast cancer cells. The MTT assays showed that 10-6 M TAM significantly reduced cell viability and altered cellular morphology. Indirect immunofluorescence microscopy revealed that galectin-3 was present on the cell surface and in the nucleus; however, its localization was diminished after 48 hours of TAM (10-6 M) exposure. Western blot analysis showed no change in galectin-3 expression at 24 hours but a significant reduction at 48 and 72 hours. In contrast, laminin expression remained unchanged. ELISA results further confirmed reduced galectin-3 but not laminin levels after 48 hours of TAM treatment. These findings suggest that galectin-3, but not laminin, is involved in TAM-induced suppression of MCF-7 cell growth and may serve as a biochemical marker for TAM responsiveness.

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major hepatic complication of obesity with a complex pathogenesis. This study aims to investigate the combined effects of empagliflozin (EMPA) and resveratrol (RSV) treatment on obesity-associated MASLD in a high-fat diet (HFD)-fed mouse model. C57BL/6J mice were fed by HFD for 23 weeks, along with treatment with EMPA (0.01% w/w), RSV (0.4% w/w), and their combination. Metabolic parameters, glucose tolerance, hepatic steatosis, inflammation, and adipose tissue morphology and function were evaluated using biochemical assays, histology, immunofluorescence, qPCR, western blotting, and 18F-FDG PET scan. Combination therapy reduced body weight gain and improved glucose tolerance more effectively than monotherapies. It significantly alleviated hepatic steatosis, as shown by decreased liver weight, triglyceride content, MASLD activity score (MAS), and serum ALT/AST levels. Co-treatment also reduced hepatic inflammation by suppressing macrophage infiltration, shifting macrophages toward an M2 anti-inflammatory phenotype, inhibiting NF-κB activation, and enhancing AMPK phosphorylation. Furthermore, the combination more effectively activated brown adipose tissue thermogenesis and induced browning in subcutaneous white adipose tissue, evidenced by upregulation of UCP1, PGC-1α, TFAM, and TMEM26, increased mitochondrial biogenesis, and enhanced cold tolerance. The combination of EMPA and RSV exerts a multi-targeted therapeutic effect against HFD-induced MASLD. This combination represents a promising novel strategy for MASLD prevention.

Inflammation-associated diseases in pigs impair the intestinal tract and impair its function, thereby posing a major threat to their health and productivity. However, current in vitro cell models fail to fully replicate porcine physiological conditions. In this study, we aimed to develop a novel inflammation model using porcine small intestinal organoids derived from adult jejunum stimulated with lipopolysaccharide (LPS). The organoids were exposed to LPS to induce inflammation, and the responses were evaluated by quantifying cytokine mRNA expression, assessing intestinal epithelial barrier integrity, and analyzing changes in the expression and localization of proteins involved in nutrient transport, receptor signaling, and enteroendocrine function using quantitative real-time PCR and immunofluorescence staining. Transcriptome analysis was performed to identify LPS-responsive differentially expressed genes and enriched pathways in porcine small intestinal organoids. LPS stimulation effectively induced inflammation in porcine small intestinal organoids, as evidenced by increased TLR4 and cytokine gene expression, compromised epithelial barrier integrity, and altered expression and localization of proteins involved in nutrient transport, bile acid signaling, and enteroendocrine function. Furthermore, transcriptome profiling revealed LPS-induced broad transcriptional changes in porcine small intestinal organoids, including enrichment of immune- and inflammation-related pathways, as well as functional categories associated with barrier maintenance, membrane function, and adaptation to inflammatory stress. Collectively, these findings demonstrate that porcine small intestinal organoids provide a physiologically relevant in vitro model for studying intestinal inflammation. This model offers a valuable platform for investigating the mechanisms underlying porcine inflammatory bowel diseases and for screening novel therapeutic agents, supporting the 3Rs principle by potentially reducing reliance on animal testing in translational research.

Oncogenesis of anal high-grade squamous intraepithelial lesions (HSIL) is highly variable. Therefore, all HSIL are ablated, leading to overtreatment and associated burden because not all lesions will progress to cancer. This exploratory study investigated differences in the tumor immune microenvironment between regressive and progressive HSIL in people living with HIV to enable a more tailored approach. Multiplex imaging mass cytometry showed more inflammation in HSIL that progressed to cancer and cancer samples, compared with HSIL that spontaneously regressed and controls. Densities of HLA-DR+ macrophage phenotypes were higher in regressive HSIL, while progressive HSIL showed a predominance of CD163+ and/or CD204+, HLA-DR- macrophage phenotypes. Validation with immunofluorescence confirmed this phenotypical distribution. Moreover, HSIL lesions closest to progression, and HSIL in individuals with a low nadir CD4 count, showed more unfavorable macrophage profiles. These preliminary findings may support the development of biomarkers or immunotherapeutic targets, enabling more individualized treatment approaches for anal HSIL.