Search Results (242)

Background and Objectives: Immunohistochemistry (IHC) is a keystone in gastric cancer (GC) management, allowing treatment customization, including for advanced or metastatic diseases. This review aims to evaluate the critical role of IHC markers, analyzing their efficiency in molecular subclassification and prediction of response to gastric cancer-targeted therapies, while also describing state-of-the-art IHC techniques and perspectives. Results: The major challenges for the GC management were structured in two main sections, as follows: (i) the current paradigm of gastric neoplasia diagnosis, which includes subsections related to the methodological and morphological foundations, the epidemiological dynamics, and risk factors, as well as differential diagnosis of poorly differentiated tumors; and (ii) the progress in 3,3′-diaminobenzidine (DAB) application and advanced reagents in gastric cancer immunohistochemistry. Discussion: Considering the role of IHC and DAB, the following topics were successively addressed in seven sections: GC key biomarkers, such as human epidermal growth factor receptor 2 (HER2), programmed death-ligand 1 (PD-L1), and DNA replication mismatch repair (MMR) system, allow direct correlation between tissue morphology and protein expression; intestinal and gastrointestinal differentiation markers; emerging and aggressive histological subtypes; epithelial–mesenchymal transition, E-cadherin, and the process of tumor budding; implementation of innovative procedures in gastric cancer immunohistochemistry; and automation, quality control, and sustainability in the pathology laboratory. Perspectives: The main directions were focused on the integration of artificial intelligence (AI) algorithms for digital quantification of the IHC signal and also on the expansion of panels to new targets, such as Claudin 18.2 (CLDN 18.2), which redefines treatment approaches in advanced stages. Conclusions: Although faced with technical and biological limitations, immunohistochemistry remains indispensable in modern gastric oncology. The evolution towards digital pathology and the refinement of scoring criteria will transform IHC from a complementary test into a visual tool that is essential for personalizing oncological treatment. Full article

This study investigated the effects of dietary lysophospholipids on growth performance, hepatic lipid metabolism, intestinal health, and dietary lipid levels of largemouth bass. The 56-day experiment included five groups: CON (0% lysophospholipids), LL50 (0.05% lysophospholipids), LP50 (0.05% lysophospholipids—0.5% oil), LP100 (0.1% lysophospholipids—1.0% oil), and LP200 (0.1% lysophospholipids—2.0% oil), with 3 replicates (30 fish/replicate) per group. The results showed that compared with the CON group, dietary supplementation of 0.05% lysophospholipid had no significant effect on the growth performance of largemouth bass, but increased the crude protein content and decreased the crude lipid content in the whole body. An amount of 0.05% lysophospholipid improved hepatic lipid utilization efficiency. Specifically, this supplementation level promoted serum lipid transport (increased serum HDL-C content and decreased triglyceride and LDL-C contents), and enhanced hepatic lipid metabolism by regulating the expression of lipid metabolism-related genes (fas, hsl, and acc) and the levels of lipid metabolites (phosphatidylcholine and fatty acids), thereby reducing hepatic triglyceride content. In addition, 0.05% lysophospholipid improved intestinal health by increasing lipase activity and intestinal villus height, up-regulating the expression of the anti-inflammatory gene (tgf-β1) and tight junction protein genes (claudin-1, claudin-4, and zo-1), and down-regulating the expression of the pro-inflammatory gene (tnf-α). In terms of dietary lipid reduction, supplementation with 0.1% lysophospholipid allowed a 1% reduction in dietary lipid level without affecting the growth performance of largemouth bass, whereas at the same level of lysophospholipid supplementation, a 2% reduction in dietary lipid level resulted in decreased growth performance of largemouth bass. These findings provide theoretical support for the practical application of lysophospholipids, and demonstrate that reducing dietary lipid inclusion by adding lysophospholipids helps to reduce feed costs and improve aquaculture economic benefits. Full article

This study aims to investigate the effects of low-energy diets (LE) supplemented with Lactobacillus reuteri postbiotics (HSY) on growth performance and intestinal health of broiler chickens. A total of 2400 one-day-old Ross 308 broiler chicks with an average initial body weight of 46.10 ± 0.04 g were randomly assigned to a 2 × 2 factorial arrangement of treatments with 12 pens and 50 broiler chickens/pen for 39 days. Treatments were (1) CTR (basal diet), (2) LE (CTR-70 kcal ME/kg), (3) HSY (CTR + 0.5 kg/t HSY), and (4) LEHSY (LE + 0.5 kg/t HSY). LE increased the feed conversion ratio (FCR) of broilers (p = 0.03) without altering ADG, ADFI, and final BW. Supplementation with HSY significantly reduced the FCR of broilers (p = 0.001). HSY upregulated the activities of amylase and trypsin in jejunal digesta (p < 0.01). Furthermore, LE upregulated the expression of intestinal barrier-related genes such as Mucin-2, Claudin-1 and Occludin, and HSY upregulated the expression of Claudin-1 (p < 0.05). LE upregulated the expression of nutrient transport carriers such as SGLT1 and TRPV6 (p < 0.01), and HSY upregulated the expression of TRPV6 (p < 0.01). LE upregulated the expression of immune-related genes such as MHC-II (p = 0.002), and HSY upregulated the expression of IFN-γ, IL-10, and TGF-β (p < 0.05). LE and HSY both downregulated the expression of intestinal lipid metabolism-related genes like ACC, while upregulating the expression of FABP4 (p < 0.05). 16S rRNA sequencing showed that the HSY increased the Chao1 index of the jejunal microbiota and enriched beneficial bacteria such as Lactobacillus salivarius and Lactobacillus avium. LE and HSY both increased the concentrations of propionic and butyrate (p < 0.05). In summary, HSY can improve gut health and mitigate the negative impact of low-energy treatment on broiler growth performance by increasing the content of endogenous enzymes in the jejunum, improving gut microbiota structure, and increasing the content of short-chain fatty acids in the jejunum. Full article

The mycotoxin deoxynivalenol (DON) is a common contaminant found in swine diets, causing decreased growth performance and poor health. Additionally, F18 enterotoxigenic E. coli is a leading cause of post-weaning diarrhea. Nursery pigs are often exposed to each of them after weaning; however, it is unknown what impact the combination of these stressors has on gastrointestinal health. Therefore, the objective of this study was to investigate the effect of pre-exposure to DON on the response of intestinal porcine epithelial cells (IPEC-J2) to challenge with enterotoxigenic F18 E. coli. Four groups were compared: Control (untreated cells), DON (cells treated with 0.5 μM DON for 24 h), F18 E. coli (multiplicity of infection 5:1, varied duration) and DON + E. coli (DON treatment with subsequent E. coli infection). Gene expression of IL-8, IL-6 and TNFα was significantly increased in cells infected with E. coli for 3 h vs. uninfected cells (p < 0.0001, p < 0.0001 and p < 0.0001, respectively). There was an interactive effect between DON and E. coli on IL-8 gene expression; cells pretreated with DON before E. coli infection had a higher expression of IL-8 than those not pretreated (p < 0.05). The concentration of IL-8 protein was significantly increased by E. coli (p < 0.0001). Claudin 1 and Occludin protein abundance were reduced by E. coli as measured by Western blot. Cytotoxicity was increased by E. coli vs. Control (p < 0.05). Pretreatment with DON increased the amount of E. coli that adhered to IPEC-J2 cells (p < 0.01) 30 min post-infection. FITC-dextran passage was increased in the DON + E. coli treatment vs. E. coli alone (p < 0.0001). Transepithelial electrical resistance (TEER) was decreased by DON when compared to untreated cells at 0 h (p < 0.0001). Similarly, DON + E. coli exhibited lower TEER vs. E. coli alone at 2 h post-infection (p < 0.0001). Taken together, these results indicate that DON pre-exposure increased the severity of E. coli infection on endpoints such as barrier permeability and E. coli adhesion. Full article

Reducing the dependence on traditional protein sources, and decreasing feeding costs and nitrogen emissions, are important tasks in livestock production. A 5 × 5 Latin square nitrogen balance trial (five castrated male pigs) and an animal growth experiment with 120 Landrace × Rongchang pigs were performed and randomly divided into five diets: a normal crude protein level diet (CON); LP diet; diversified LP containing broken rice, rapeseed meal, and DDGS (DLP); DLP + 0.05% cellulase (DLP + CE); and DLP + 20% fermented feed (FDLP). The CON group showed higher nitrogen intake, urinary nitrogen, and total nitrogen excretion than the other four groups (p < 0.05). The fecal nitrogen was decreased with the LP, DLP + CE, and FDLP groups compared to the CON group (p < 0.05). The mRNA expression of jejunal fatty acid transport protein 1 was upregulated in the LP, DLP + CE, and FDLP groups compared to the CON and DLP groups (p < 0.05). The DLP + CE group showed a higher intramuscular fat content in pigs than the CON and DLP groups (p < 0.05). In the LD muscle, the FDLP and DLP + CE groups upregulated fatty acid synthase expression compared to the LP and DLP groups (p < 0.05). Colonic mRNA expression of zonula occludens-1 and claudin-1 was upregulated in the FDLP group compared to the CON and DLP groups (p < 0.05). These results suggest that the supplementation of cellulase and fermented feed in DLP diets improved nitrogen utilization and intestinal health without compromising growth performance or meat quality in Landrace × Rongchang pigs. Full article

Background: The interaction between keratinocytes and proinflammatory cytokines is essential in the development of psoriatic lesions. The synergism among these cytokines and their involvement in ferroptosis are not yet elucidated. This study aimed at evaluating the early impact of a complete proinflammatory microenvironment on keratinocyte differentiation, intercellular adhesion, proliferation, and induction of ferroptosis. Methods: HaCaT cells were differentiated with 1.8 mM CaCl₂ and treated with a cytokine combination (MIX) containing IL-17A, IL-22, IL-23, and TNF-alpha for 24 and 48 h. Claudin 1 (CLDN-1), Zonula Occludens 1 (ZO-1), and keratins (K)10/K14 expression was analyzed by immunofluorescence and immunoblot analysis, paralleled by proliferation and ultrastructural analysis. Ferroptosis was induced with erastin and RSL3 and evaluated by testing glutathione (GSH)/glutathione peroxidase 4 (GPX4) protein expression, GSH levels, cell availability/toxicity, intracellular iron and ATP levels. Results: After MIX incubation at T48, CLDN-1 and ZO-1 immunofluorescences were reduced in HaCaT cells, while K10 and K14 were unaffected. The proliferative activity was reduced. Psoriatic-like MIX triggered the ferroptotic pathway, as shown by the increase in intracellular iron levels as well as by the reduction in GPX4 protein expression, the decrease in GSH levels, cell availability, and ATP levels. Conclusions: This experimental model mimics the early pathogenetic processes underlying psoriatic plaque formation/progression paving the way for new therapeutic strategies. Full article

Avian pathogenic Escherichia coli (E. coli) impairs poultry production and causes substantial economic losses. This study investigated the effects of Lactobacillus reuteri postbiotics (LR) on growth performance and intestinal health of broiler chickens challenged with E. coli. A total of 180 one-day-old Arbor Acres+ broilers were allocated into three groups (six replicates per group and 10 chicks each replicate): CTR, control group; E. coli-infected group, orally challenged with a mixture of E. coli O₁, O₂, and O₇₈ at a dose of 10⁹ CFU/mL; LR + E. coli-infected group, challenged with E. coli and fed a basal diet supplemented with 100 mg/kg LR. The results showed that dietary LR significantly improved the average daily gain (ADG) in the LR + E. coli group compared to the E. coli-infected group from days 1 to 18 (p < 0.05). However, no significant differences in average daily feed intake (ADFI) or feed conversion ratio (FCR) were observed among the CTR, E. coli, and LR + E. coli groups. Infection with E. coli led to lower total antioxidant capacity in jejunum and activity of total superoxide dismutase in ileum. Moreover, dietary LR significantly alleviated the down-regulation of Mucin2 and Aquaporin-3 gene expression in jejunum and ileum caused by E. coli infection and up-regulated the gene expression of Claudin-1 and Zonula occludens 1 in the ileum. In addition, dietary LR treatment led to the up-regulation of interleukin-10 mRNA transcripts in the jejunum. Further analysis demonstrated that dietary supplementation with LR reshaped the ileal flora of birds challenged with E. coli via elevating the relative abundance of Romboutsia and Bacteroidota, while reducing the abundance of Candidatus_Arthromitus and Escherichia-Shigella. In conclusion, dietary LR supplementation improved the expression of intestinal barrier and anti-inflammatory genes and reshaped the intestinal flora in E. coli-infected broilers. Full article

Background: Skin barrier dysfunction leads to increased transepidermal water loss (TEWL), inflammation, and compromised skin protection. While Paeonia ostii (peony) flowers are recognized in traditional Chinese medicine for their reducing melanin synthesis, anti-inflammatory, and anti-aging effects, their role in repairing skin barrier damage has not been fully explored. Methods: We investigated the therapeutic potential of peony flower extract (PFE) in the tape-stripping-induced mouse model with skin barrier damage. Skin surface imaging, hydration measurements, H&E, proteomics, qPCR, and immunofluorescence were applied to clarify the potential mechanism of PFE in attenuating skin barrier impairment. Results: PFE significantly reduced erythema, TEWL, and edema while restoring epidermal architecture. Proteomics analysis identified cornified envelope formation and tight junction assembly as essential mechanisms in skin barrier repair. It increased the expression of key skin barrier proteins, including filaggrin (Flg), involucrin (Ivl), loricrin (Lor), claudin-1 (Cldn1), tight junction protein 1 (Tjp1), and occludin (Ocln). Conclusions: This study demonstrates that PFE restores skin barrier integrity by upregulating key structural proteins within the cornified envelope and tight junction. These findings suggest that PFE is a promising therapeutic candidate for skin barrier repair, with high potential in translational medicine applications. Full article

The effects of chlorogenic acid (CGA) on intestinal histomorphology, barrier integrity, antioxidant parameters, and gut microbiota in heat-stressed rabbits were assessed in this study. One hundred and twenty weaned New Zealand rabbits were assigned to three groups: control (CON) at 25 ± 1 °C, heat stress (HS) at 35 ± 1 °C, and HS with CGA supplementation (HS + CGA) at 35 ± 1 °C. Rabbits in the CON and HS groups were fed a basic diet, while those in the HS + CGA group receive the basic diet added with 800 mg/kg CGA. HS induced intestinal oxidative stress, impaired intestinal morphology and barrier function, and altered the gut microbiota. CGA supplementation mitigated HS-induced increases in serum diamine oxidase and D-lactate levels, and intestinal malondialdehyde content (p < 0.05), and countered HS-induced reductions in intestinal superoxide dismutase activity, villus height/crypt depth ratio, and claudin-1 and ZO-1 mRNA expressions (p < 0.05). In addition, HS decreased the abundances of Akkermansia and uncultured_bacterium_g__Akkermansia and increased the Firmicutes/Bacteroidota ratio and uncultured_bacterium_g__unclassified_o_Clostridia_UCG-014 abundance as well as the abundance of bacterial functions related to animal_parasites_or_symbionts and human_pathogens_all. HS-induced gut microbiota dysbiosis was significantly restored by CGA supplementation. The findings indicated that dietary 800 mg/kg CGA supplementation effectively safeguarded intestinal health in rabbits under high temperatures. Full article

Background: Infection with Mycobacterium avium paratuberculosis (MAP) is closely associated with Crohn’s disease (CD) development, where excessive inflammation and marked intestinal damage are observed. Objectives: In this study, the role of short chain fatty acids, including propionic acid (PPA) and butyric acid (BA), was evaluated in an in vitro model, mimicking CD characteristics. Methods: MAP-infected THP-1 macrophages were treated with 1 mM and 10 mM of PPA or BA, and the conditioned media was co-cultured in Caco-2 cells. Results: Both PPA and BA caused an M2 shift with significant downregulation (p-value < 0.0001) in pro-inflammatory markers at both the RNA and protein levels. The downregulation is most likely due to the antimicrobial properties of PPA and BA. MAP growth was inhibited by several folds in MGIT (Mycobacteria Growth Indicator Tube) culture media supplemented with PPA or BA. Dysfunctional Caco-2 intestinal epithelial cells’ integrity and function, due to MAP infection, were restored with PPA and BA treatment. Specifically, NOX1 expression was significantly decreased in 10 mM of PPA or BA-treated cells (p < 0.001), as validated by RT-PCR and microscopy. PPA and BA restored tight junction integrity by decreasing Claudin-2 expression in the MAP group. Conclusions: The data clearly demonstrated that short chain fatty acids contain anti-inflammatory and antimicrobial properties with downstream beneficial effects on damaged intestinal epithelial cells, suggesting potential benefits as a dietary supplement for CD patients, particularly those who are not pregnant, due to a possible increased risk of autism spectrum disorder (ASD) development in offspring associated with propionic acid exposure. Full article

Background and Clinical Significance: Zolbetuximab, a claudin 18.2-targeted monoclonal antibody, has demonstrated efficacy in advanced gastric cancer. Hypoalbuminemia has emerged as a notable adverse effect, but its underlying mechanism remains unclear. Case Presentation: A 53-year-old male with unresectable advanced gastric cancer received zolbetuximab-based combination therapy, achieving tumor regression enabling conversion surgery. During six cycles of treatment, serum albumin levels decreased from 4.3 g/dL to 3.5-3.6 g/dL (grade 1 hypoalbuminemia). A histopathological examination of the surgical specimen revealed hypertrophic gastritis characterized by marked foveolar hyperplasia, increased mucus secretion, and pyloric gland metaplasia on the lesser curvature. These findings suggest that zolbetuximab-induced mucosal alterations may contribute to hypoalbuminemia through enhanced protein loss. Conclusions: This is the first pathological documentation of hypertrophic gastritis associated with zolbetuximab therapy. Clinicians should monitor albumin levels during treatment and consider nutritional support when indicated. These findings provide important insights for optimizing patient management and ensuring safe conversion surgery planning. Full article

This study aimed to investigate the effects of glycerol monolaurate (GML) on the growth performance, intestinal morphology, inflammatory factors, and gut microbiota of weaned piglets challenged by lipopolysaccharide (LPS). A total of eighteen weaned piglets [Duroc × (Landrace × Yorkshire), average weight 7.54 ± 0.86 kg] were randomly assigned to the following three groups: (1) CON:basal diet; (2) LPS: basal diet + LPS challenge; and (3) LPS_GML: with 1000 mg/kg GML. On day 21, LPS or saline was injected into the piglets via intraperitoneal injection, and samples were collected after 4 h. Results showed that no differences were observed in the average daily gain (ADG), average daily feed consumption (ADFI), and G/F ratio. GML supplementation increased (p < 0.05) the villus height and villus height/crypt depth ratio, the protein expression of claudin-1 and occludin, the mRNA expression of claudin-1, SOD, and GSH-Px, while decreased (p < 0.05) the serum diamine oxidase (DAO) and reactive oxygen (ROS) concentrations and the mRNA expression of IL-1β, IL-6, and IL-8 compared with the LPS group. Moreover, GML regulated LPS-induced gut microbiota dysbiosis by reshaping the composition of gut microbiota, such as increasing (p < 0.05) Actinobacteriota, Lactobacillus amylovorus DSM 20531, Lactobacillus mucosae LM1, and Bifidobacterium boum, while decreasing (p < 0.05) Spirochaetota. Dietary GML supplementation improved the barrier function, reduced the inflammatory factors, and modulated the composition of gut microbiota in piglets challenged by LPS. Full article

The dedifferentiation of smooth muscle cells (SMCs) is the main cause of atherosclerosis and vascular calcification. This study integrated the gene expression data of multiple microarrays to identify relevant marker molecules. A total of 72 Gene Expression Omnibus (GEO) samples (GSM) were collected from 10 gene expression data series (GSE) and divided into five groups: non-SMC, SMC, atherosclerotic SMC (SMC-ath), calcified SMC (SMC-calc), and treated SMC (SMC-t). The SMC-t group included synthetic SMCs that had undergone treatment to inhibit proliferation, migration, or inflammation. The gene expression data were merged, normalized, and batch effects were removed before differential gene expression (DGE) analysis was performed via linear models for microarray data (limma) and statistical analysis of metagenomic profiles (STAMPs). The genes with expressions that significantly differed were subsequently subjected to protein-protein interaction (PPI) and functional prediction analyses. In addition, the random forest method was used for classification. Twelve proteins that may be marker molecules for SMC differentiation and dedifferentiation were identified, namely, Proprotein convertase subtilisin/kexin type 1 (PCSK1), Transforming growth factor beta-induced (TGFBI), Complement C1s (C1S), Phosphomannomutase 1 (PMM1), Claudin 7 (CLDN7), Calcium binding and coiled-coil domain 2 (CALCOCO2), SAC3 domain-containing protein 1 (SAC3D1), Natriuretic peptide B (NPPB), Monoamine oxidase A (MAOA), Regulator of the Cell Cycle (RGCC), Alpha-crystallin B Chain (CRYAB), and Alcohol dehydrogenase 1B (ADH1B). Finally, their possible roles in SMCs are discussed. This study highlights the feasibility of bioinformatics analysis for studying SMC dedifferentiation. Full article

Background/Objectives: Fonte Essenziale^®, a mineral water rich in bicarbonate, sulfate, calcium, and magnesium, has shown potential in modulating the gut–liver axis and microbiota in hepatic steatosis. However, its long-term effects on intestinal permeability (IP), systemic inflammation (SI), and oxidative stress—key factors in Metabolic dysfunction-associated steatotic liver disease (MASLD)—remain unexplored. Methods: Eighty-seven MASLD patients were randomized into two groups: group A received Fonte Essenziale^® (400 mL/day, fasting) plus a controlled nutritional regimen for 12 months, followed by a 6-month water washout; group B followed only the controlled nutritional regimen. IP markers, SI (IL-1β, IL-6, TNF-α), oxidative stress (dROMs/BAP), and clinical data (including Controlled Attenuation Parameter—CAP) were assessed at baseline (T0), 12 months (T12), and post-washout (T18). Baseline increased IP (in-IP) was defined by fecal zonulin > 110 ng/mL and serum LBPp > 10 µg/mL; improvement (im-IP) required normalization of both. A ≥30% CAP reduction indicated steatosis improvement. Results: Thirty-eight patients in group A and thirty-nine in group B completed the study. At T12, group A showed significant reductions in fecal zonulin (p: 0.0163) and serum LBPp (p < 0.0001), with increased occludin and claudin 1 (all p < 0.0001). Im-IP prevalence was higher in group A (p: 0.0037). Group A also showed significant reductions in IL-1β, TNF-α, IL-6, LPS, and dROM/BAP (all p < 0.05), especially among those with im-IP. CAP, insulin, and HDL levels improved significantly (all p < 0.0001). Multivariate analysis confirmed water intake (aOR: 2.185, p: 0.001) and im-IP achievement (aHR: 1.267, p: 0.021) as predictors of steatosis improvement. Benefits persisted at T18. Conclusions: Prolonged Fonte Essenziale^® intake improved hepatic steatosis and MASLD outcomes by modulating IP, SI, and oxidative stress. This trial has been registered on clinicaltrials.gov (NCT07211113). Full article

As an economically important species endemic to the upper tributaries of Yangtze River in China, long-finned gudgeon fish (Rhinogobio ventralis) has been classified as endangered due to habitat destruction and population decline. In this study, we constructed a chromosome-level genome assembly of R. ventralis by integration of MGI, PacBio and Hi-C sequencing technologies. The final genome assembly was 1015.9 Mb in length (contig N50: 25.91 Mb; scaffold N50: 39.99 Mb), and 97.19% of the haplotypic genome sequences were anchored onto 25 chromosomes. Repetitive elements accounted for 51.00% of the entire genome assembly. A total of 23,220 protein-coding genes were predicted for the assembled genome, of which 99.79% were functionally annotated. Genome evaluation revealed 99.72% completeness for the genome assembly. Through genome-wide prediction of antimicrobial peptides (AMPs), we identified and localized 561 putative AMP-containing genes in the R. ventralis genome. These genes were further classified into 185 distinct functional categories based on public databases, with the top ten components of Penetratin (21.74%), Histone (5.70%), E6AP (4.09%), Scolopendin 1 (2.67%), D38 (2.31%), WBp-1 (2.13%), Defensin (2.13%), Claudin 1 (1.96%), Azurocidin (AZU1, 1.78%), and Ubiquitin (1.60%). Our data presented here provide a potential genetic resource for promoting fundamental research and wild population conservation of this endangered fish species. Full article