The strategic focus on improving the quality of eggs, which are foundational and nutritious foods, offers a straightforward method to elevate the nutritional value of diets. However, studies on strategies to improve egg quality remain limited. Lycopene, a lipophilic carotenoid derived from red fruits and vegetables, enhances growth performance, lowers blood lipid levels, and improves antioxidant capacity. Therefore, in this study, a total of 360 laying hens were randomly divided into four groups. The CON group was fed a basal diet, the LYC group was supplemented with 120 mg/kg lycopene, whereas the CCBA group was supplemented with 60 mg/kg β-carotene, 250 mg/kg curcumin, 250 mg/kg allicin, and 500 mg/kg sodium butyrate, the CLYC group replaced β-carotene with lycopene, while other ingredients remained the same. For the yolk score, the LYC group significantly increased at days 14, 21, 28, 35, and 42 compared to the CON group (p < 0.05). The UFA and Vitamin A contents were significantly elevated in the LYC group compared to the CON group (p < 0.05). The result of lipid metabolism genes expression showed that the relative expression of ACC, LXR-α, and PPAR-α significantly increased in the CLYC and CCBA groups compared with the CON group (p < 0.05), the relative expression of LXR-α in the LYC group remarkably higher than that in the CON group (p < 0.05). Additionally, heatmap analysis showed that Turicibacter and Veillonella were enriched in the LYC group. The results of this study indicated that lycopene supplementation significantly improved egg quality, hepatic lipid metabolism, and systemic immunity in laying hens. Furthermore, analysis of gut revealed that lycopene increased ileal goblet cell numbers and IgA production, while enriching beneficial microbiota.

β-carotene, a carotenoid with strong antioxidant and immunity-enhancing properties, is a safe and widely available precursor of vitamin A in nature. Pumpkin powder, as a familiar agricultural product, is rich in vitamins, amino acids, and minerals, which also contain β-carotene. Currently, there remain few published reports on the effects of pumpkin powder on antioxidant function, reproductive performance, and egg quality in laying hens, representing a significant gap in existing research. 270 laying hens were divided into three groups; the CON group was fed a basal diet, the BCA group was fed the basal diet supplemented with 120 mg/kg β-carotene, and the PPD group was fed a diet containing pumpkin powder to provide an equivalent dose of β-carotene. On 35 days, the β-carotene content in the egg yolk was significantly increased in the BCA and PPD groups compared to the CON group (p < 0.01). On 49 days, the UFA, vitamin A, and β-carotene content in the BCA and PPD groups were significantly higher than in the CON group (p < 0.01). In the BCA and PPD groups, levels of SOD, CAT, GSH-Px, and T-AOC markedly increased in yolk and serum. The relative expression of Keap1 and Nrf2 levels in the BCA and PPD groups was remarkably higher compared to the CON group (p < 0.01). The relative expression levels of the BMP15, and FOXL2 genes in the BCA and PPD groups of laying hens were significantly higher than in the CON group (p < 0.05). The relative abundance of the genera Megamonas and Eubacterium in the fecal microbiota was significantly higher in the PPD group than in the CON group (p < 0.05). In summary, dietary supplementation with pumpkin powder as a source of β-carotene in laying hens effectively enhanced egg production and quality, improved systemic antioxidant status and gut health, and reduced ovarian inflammation.

Corn gluten meal (CGM) and wheat bran are widely recognized as two major by-products generated by the food-processing industry. The corn gluten meal-wheat bran mixture (CWM) was fermented using Aspergillus niger and Lactobacillus fermentum, and the potential of ultrasound-assisted fermentation to enhance its value was evaluated. Five fermented corn gluten meal-wheat bran mixtures (FCWMs) were prepared by applying ultrasound at 0, 130, 195, 260, and 320 W for 5 min. The results indicated that ultrasound-assisted fermentation at 195 W yielded the most favorable effects. At this power level, microbial biomass was increased, while high-molecular-weight proteins were effectively degraded, resulting in substantial increases in trichloroacetic acid-soluble protein (TCA-SP) and total free amino acid (TFAA) contents. Furthermore, the levels of beneficial metabolites, including total phenolics, flavonoids, and carotenoids, were elevated. Concurrently, antinutritional factor levels were reduced, and antioxidant capacity was enhanced.

Background: Methotrexate (MTX) effectively eliminates cancerous cells but can also cause inflammation intestinal, known as mucositis. Forsythiaside A (FTA) from Forsythia suspensa has shown promise in relieving mucositis by targeting the NLRP3 pathways. Since NLRP3 inflammasome activation is negatively regulated by autophagy, this study explores how FTAmediated autophagy affects NLRP3 inflammasome in treating MTX-induced intestinal inflammation. Method: Intestinal mucositis was induced in rats with MTX. FTA's impact was assessed using HE staining and ELISA. The mechanism was studied using immunofluorescence, western blot, and ELISA. result: FTA treatment resulted in reduced levels of D-lactic acid and diamine oxidase (DAO) in MTX-treated rats. Western blot and immunofluorescence analyses revealed up-regulation of Beclin- 1 and LC3II/I, accumulation of LC3, and down-regulation of p62 expression levels in MTXtreated rats following 40 or 80 mg/kg FTA intervention. However, when the autophagy inhibitor 3-MA was used, the intestinal pathology was exacerbated, the inflammatory scores increased, and serum levels of TNF-α, IL-1β, and IL-18 were elevated. Western blotting indicated decreased LC3II/I expression, while NLRP3, cleaved caspase 1, and cleaved IL-1β expressions were upregulated. Conclusion: These findings suggested that FTA alleviated MTX-treated intestinal mucositis by activating autophagy, which in turn inhibits the NLRP3 inflammasome.

Corn gluten meal and rice bran mixture (CRM) are major agricultural by-products of food processing. The nutritional quality of fermented CRM (FCRM) was enhanced through ultrasound-assisted fermentation with Aspergillus oryzae AOX and Lactobacillus helveticus LHS. Additionally, polydopamine/chitosan-gum Arabic/FCRM (P-C-FCRM) microcapsules were fabricated under ultrasonic conditions to enhance the stability of bioactive compounds. After ultrasound-assisted fermentation, high-molecular-weight proteins, zein, and fibers in FCRM were significantly degraded, resulting in 3.32-, 1.42-, and 11.87-fold increases in peptides, total free amino acids, and reducing sugars, respectively, while levels of bioactive compounds-including phenolics, flavonoids, and carotenoids-were also markedly elevated. P-C-FCRM achieved 95.47% encapsulation efficiency and exhibited excellent storage stability. In vitro simulated digestion experiments demonstrated that the degradation rate of unencapsulated FCRM was 59.17%, whereas that of P-C-FCRM was 20.31%, enabling the targeted release of P-C-FCRM in the intestine. Furthermore, in vivo experiments were conducted to examine the regulatory effects of P-C-FCRM on the gut microbiota of chicks. In vivo studies demonstrated that P-C-FCRM improved chick growth and gut health by increasing probiotics and short-chain fatty acids. This work introduces a scalable bioprocess integrating ultrasound, microbial fermentation, and microencapsulation, offering a novel strategy for the valorization of agricultural by-products into value-added functional feed products.

BackgroundMaternal dietary intervention utilizing complex additives rich in β-carotene has demonstrated the capacity to enhance embryonic intestinal development and influence microbial composition in offspring. Nevertheless, the extended impact of maternal β-carotene inclusion on the intestinal health of post-hatching chicks is still not fully elucidated.ObjectiveThis research aimed to evaluate the impacts of maternal β-carotene supplementation on the intestinal development and microbial communities in chicks after hatching.MethodsA total of 1,215 Hy-Line Brown laying hens were divided into three groups, with each group containing five subgroups of 81 hens each. The hens received a basal diet (CON) or diets added with β-carotene at 120 mg/kg (LBC) or 240 mg/kg (HBC). After 6 weeks of dietary intervention, eggs were collected for incubation. Following hatching, 40 male chicks from each group were randomly chosen and evenly distributed into five distinct subgroups. All chicks were then reared on a uniform basal diet for a duration of 6 weeks.ResultsChicks from the LBC group exhibited significantly higher initial body weight, enhanced villus height (VH), greater villus height to crypt depth ratio (VCR), higher goblet cell (GC) density, and elevated expression of Mucin 2 (MUC2), zonula occludens-1 (ZO1), and zonula occludens-2 (ZO2) in both the jejunum and ileum at 42 days of age. In addition, maternal inclusion of β-carotene 240 mg/kg markedly improved VCR in the jejunum and ileum of chicks. In spite of the observed enhancements in intestinal health, there were no notable variations in overall growth performance across the groups during days 1 to 42. Beta diversity analysis revealed distinct microbial clustering in the ileum of both LBC and HBC groups, clearly separated from the CON group. Taxonomic profiling showed an enrichment of Marivita, Burkholderia, and Turicibacter in the CON group; Oxalobacteraceae, Anoxybacillus, Roseburia, and Anaerorhabdus in the LBC group; and SMB53 and Allobaculum in the HBC group. Correlation analyses indicated positive associations between Anaerorhabdus, Anoxybacillus, and Roseburia and improved intestinal histomorphology, GC abundance, and barrier-related gene expression.ConclusionThese findings suggest that maternal β-carotene addition confers sustained benefits to intestinal health in offspring, potentially mediated by modulation of the cecal microbiome up to day 42.

In poultry, maternal nutritional interventions affect the development and intestinal microbiota of embryos. β-carotene possesses immune-boosting and gut microbiota-regulating properties. We examined the influences of supplementing hen diets with β-carotene on offspring growth, development, and immunity to determine whether maternal β-carotene benefits offspring health. Our findings showed that β-carotene increased serum IgG, lysozyme, and beta-defensins in hens, subsequently elevated these parameters in the serum of their offspring, and promoted their growth and development. In offspring, there were significant positive correlations between body weights and intestinal development indices with serum lysozyme and beta-defensin levels. The augmentation of vertical transfer of lysozyme and beta-defensins may be linked to the increased expression of these genes in the maternal jejunum. The number of shared taxa between the magnum and offspring gut is higher than that between the maternal gut and offspring. Among the taxa, were increased in the maternal magnum and gut microbiome, only the Caloramator abundance was significantly elevated in the guts of 21-day-old offspring. In conclusion, maternal β-carotene inclusion improves offspring growth and development, potentially through enhancing maternal intestinal immunity and thereby promoting immune-mediated maternal effects. The vertical transfer of maternal microbes to offspring exhibits selectivity in chicken.

The corn gluten meal-corn stalk mixture (CCM) was used as a fermentation substrate composed of two agricultural by-products: corn gluten meal, obtained during wet milling for starch and syrup production, and corn stalk, a lignocellulosic residue left after corn harvest. To facilitate the valorization of agricultural by-products, ultrasound-assisted fermentation was conducted using Aspergillus niger and Limosilactobacillus fermentum at power densities of 50 W/L (F-L) and 100 W/L (F-H), respectively. CCM ultrasound-assisted fermentation led to significant degradation of macromolecules and zein. Compared to the non-ultrasound group, trichloroacetic acid-soluble protein in F-H increased by 6.79-fold, and total amino acids increased by 1.44-fold. Total phenols and flavonoids in F-L and F-H increased, thereby enhancing the antioxidant capacity of CCM. Ultrasound-assisted fermentation primarily increased the abundance of Limosilactobacillus, Pediococcus, Pichia, and Aspergillus, while reducing the abundance of Pantoea, Xanthomonas, Curtobacterium, Staphylococcus, Alternaria, Blumeria, Cladosporium, Fusarium, and Mucor. Environmental acidification drove the microbial community to shift towards more acid-tolerant species. Functional predictions revealed upregulated pathways involved in amino acid and bioactive substance biosynthesis, providing a new perspective on the high-value utilization of CCM.

The strategic focus on improving the quality of eggs, a foundational and nutritious food, offers a straightforward method to elevate the nutritional value of diets. However, studies on strategies to improve egg quality remain limited. Lycopene, a lipophilic carotenoid derived from red fruits and vegetables, enhances growth performance, lowers blood lipid levels, and improves antioxidant capacity. Therefore, in this study, a total of 360 laying hens were randomly divided into four groups. The CON group was fed a basic diet, the LYC group supplemented with 120 mg/kg lycopene, whereas the CCBA group supplemented with 60 mg/kg β-carotene, 250 mg/kg curcumin, 250 mg/kg allicin, and 500 mg/kg sodium butyrate, the CLYC group replaced β-carotene with lycopene, while other ingredients remained the same. For the yolk score, the LYC group significantly increased at days 14, 21, 28, 35, and 42 compared to the CON group ( P< 0.05 ). The UFA and Vitamin content are significantly elevated in the LYC group compared to the CON group ( P<0.05 ). The result of lipid metabolism genes was shown, the relative expression of ACC , LXR-α , and PPAR-α significantly increased in the CLYC and CCBA groups compared with the CON group ( P< 0.05 ), the relative expression of LXR-α in the LYC group remarkedly higher than the CON group (P< 0.05). Additionally, the heatmap analysis showed that Turicibacter and Veillonella were enriched in the LYC group. The results of this study indicated that lycopene supplementation significantly improved egg quality, hepatic lipid metabolism, and systemic immunity in laying hens. Furthermore, analysis of gut revealed that lycopene increased ileal goblet cell amount and IgA production, while enriching beneficial microbiota.

Modern management of chicks hinders the vertical transmission of intestinal microbiota, which is closely related to immunity. Inulin is a substrate that can be utilized by the microbiota. This study aimed to determine whether fecal microbiota transplantation (FMT) combined with inulin played a "1 + 1 > 2" role in enhancing the development and function of immune organs. Chicks were treated with 1 % inulin and/or fecal microbiota suspension on days 1-6. The growth performance, immune organ development, and immune indicators were evaluated on days 7, 14, and 21. Results showed that the combination of FMT and inulin significantly increased the immune organ index on day 7 and promoted the morphological structure and the expression of proliferating cell nuclear antigen (PCNA) in immune organs on days 7, 14, and 21. Each treatment increased the gene expression of interferon-gamma (IFN-gamma), interleukin-4 (IL-4), interleukin-2 (IL-2), B cell-activating factor receptor (BAFFR), B cell linker (BLNK), C-X-C Motif Chemokine Ligand 12 (CXCL12), C-X-C Motif Chemokine Receptor 4 (CXCR4), and Biotin (Bu-1) to varying degrees. FMT combined with inulin significantly increased the expression of IgA-positive cells on days 7 and 14. In conclusion, the synergistic effect of FMT and inulin had beneficial impacts on the development and function of immune organs.

Fecal microbial transplantation (FMT) is an important technology for treating diarrhea and enteritis. Additionally, FMT has been applied to improve productivity, alter abnormal behavior, relieve stress, and reduce burdens. However, some previous studies have reported that FMT may cause stress in acceptor animals. Inulin, a prebiotic, can promote growth, enhance immunity, and balance the gut microbiota. Currently, there are limited reports on the effects of combining FMT with inulin on early growth performance in chicks. In this study, a total of 90 1-day-old chicks were randomly divided into the control group (CON), FMT group, and inulin group (INU). The CON group was fed a basic diet, whereas the FMT and INU groups received fecal microbiota transplantation and FMT with inulin treatment, respectively. Compared with the FMT and CON groups, the INU group presented significantly greater average daily gain (ADG) and average daily feed intake (ADFI) values (P < 0.05). However, the organ indices did not significantly change (P > 0.05). The ratio of the villi to crypts in the ileum significantly differed at 21 and 35 days (P < 0.05). In addition, the cecum concentrations of acetic acid and butyric acid significantly increased in the INU group (P < 0.05). In addition, gut inflammation and serum inflammation decreased in the INU group, and immune factors increased after inulin supplementation. (P < 0.05). Firmicutes and Bacteroidetes were the dominant phyla, with more than 90 Experimental design process diagram

Ochratoxin A (OTA) is a widespread environmental toxin that poses a serious threat to human and animal health. OTA has been shown to cause cellular and tissue damage and is a global public health problem. However, the effects of OTA on gastrointestinal aging have not been reported. The aim of this study was to investigate the effects of OTA on intestinal aging in vitro and in vivo. In vitro experiments showed that OTA induced cellular inflammation through calcium overload and oxidative stress, significantly up-regulated the expression of P16, P21, and P53 proteins, markedly increased senescence-associated β-galactosidase activity (SA-β-gal) positive cells, and obviously decreased the expression of proliferating cell nuclear antigen (PCNA) proteins, which led to intestinal cell senescence. Meanwhile, we found that treatment with β-carotene ameliorated OTA-induced intestinal cell senescence. Consistent with the results of the in vitro experiments, in vivo studies showed that the intestinal aging of mice fed OTA was significantly higher than that of the control group. In conclusion, OTA may induce intestinal aging through calcium overload, oxidative stress and inflammation. This study lays a foundation for further research on the toxicological effects of OTA.

Calcium ions (Ca2+) regulate cell proliferation and differentiation and participate in various physiological activities of cells. The calcium transfer protein inositol 1,4,5-triphosphate receptor (IP3R), located between the endoplasmic reticulum (ER) and mitochondria, plays an important role in regulating Ca2+ levels. However, the mechanism by which IP3R1 affects porcine meiotic progression and embryonic development remains unclear. We established a model in porcine oocytes using siRNA-mediated knockdown of IP3R1 to investigate the effects of IP3R1 on porcine oocyte meiotic progression and embryonic development. The results indicated that a decrease in IP3R1 expression significantly enhanced the interaction between the ER and mitochondria. Additionally, the interaction between the ER and the mitochondrial Ca2+ ([Ca2+]m) transport network protein IP3R1-GRP75-VDAC1 was disrupted. The results of the Duolink II in situ proximity ligation assay (PLA) revealed a weakened pairwise interaction between IP3R1-GRP75 and VDAC1 and a significantly increased interaction between GRP75 and VDAC1 after IP3R1 interference, resulting in the accumulation of large amounts of [Ca2+]m. These changes led to mitochondrial oxidative stress, increased the levels of reactive oxygen species (ROS) and reduced ATP production, which hindered the maturation and late development of porcine oocytes and induced apoptosis. Nevertheless, after treat with [Ca2+]m chelating agent ruthenium red (RR) or ROS scavenger N-acetylcysteine (NAC), the oocytes developmental abnormalities, oxidative stress and apoptosis caused by Ca2+ overload were improved. In conclusion, our results indicated IP3R1 is required for meiotic progression and embryonic development by regulating mitochondrial calcium and oxidative damage.

Using new materials to improve detection accuracy and efficiency is important to broaden the application of immunoassay sensors. Among numerous materials for improving sensors, chitosan, as a natural polysaccharide, has excellent biocompatibility, mechanical adjustability, stimulation sensitivity and porous structure, which holds significant potential for enhancing the performance of immunoassay sensors. However, at present, there are few systematic reviews on the application of chitosan in immune sensors. In this paper, the principle of immunoassay is discussed systematically, we reviewed the recent development of chitosan enhancement strategies in various immunoassay sensors, including surface plasmon resonance immunoassay sensors, colorimetric immunoassay sensors, electrochemical immunoassay sensor, electrochemical luminescence immunoassay sensors are reviewed. Focused on the theoretical basis of improving sensor performance in immunoassay by use chitosan, as well as the various functions and applications of chitosan, and discussed how to solve the challenges of immunoassay sensors by using chitosan and the future research trend. By providing a robust foundation for the development of more efficient detection platforms, it provides insights for advancing the use of chitosan in the detection of complex biological samples. This is crucial for promoting the widespread application of immunoassay sensors with high performance in clinical diagnosis, environmental monitoring and food safety.

Long-chain chlorinated paraffins (LCCPs) are industrial raw materials extensively utilized worldwide. Recently, their environmental impact has escalated, exacerbating challenges in animal husbandry and contributing to pollution from the food industry, which poses certain risks to animal growth and development. However, the toxicological effects of LCCPs exposure on poultry remain inadequately understood. The liver is a critical organ in poultry, serving not only as the largest digestive gland but also as the center of metabolism. Consequently, this study employed primary chicken embryo hepatocyte as a model to investigate the toxicological effects of LCCPs exposure and its potential mechanisms of action. Our findings indicate that the proliferation capacity of primary chicken embryo hepatocytes exposed to LCCPs at concentrations of 1, 10, and 100 μg/L was significantly diminished, with an observed arrest in the G0/G1 phase and a notable reduction in the proportion of cells in the G2/M phase. Additionally, we observed that LCCPs exposure markedly decreased the autophagy levels in primary chicken embryo hepatocytes while significantly increase the levels of apoptosis. To elucidate the molecular mechanisms underlying LCCP-induced apoptosis in these cells, we assessed oxidative stress levels (ROS) and mitochondrial membrane potential, and found that the level of ROS was significantly increased, and the level of mitochondrial membrane potential was significantly decreased in primary chicken embryo hepatocytes after exposure to LCCPs. To further clarify whether LCCPs induced apoptosis in primary chicken embryo hepatocytes through oxidative stress, oxidative stress inhibitors (NAC) were used, and it was found that apoptosis caused by LCCPs exposure was significantly alleviated. These data suggest that LCCPs exposure could induce apoptosis in primary chicken embryo hepatocytes through oxidative stress. In conclusion, the current work shows that LCCPs have multiple toxic effects on primary chicken embryo hepatocytes, and lays a theoretical foundation for future research on the harmful effects of LCCPs in the poultry industry.

Previous studies have shown that various active components of licorice have anticancer effects. However, few studies have investigated the mechanism of action of licorice in gastric cancer. The effect of active compounds in licorice on the biological activity of gastric cancer cells was investigated in vitro (MKN-45 cells). Network pharmacology and molecular docking were used to predict the potential targets of licorice against gastric cancer and verify the binding stability of target proteins to compounds. In addition, the anticancer effect of licorice was assessed using a mouse model of gastric cancer. The licorice-active component (quercetin) effectively inhibited proliferation, caused cell cycle arrest, and promoted apoptosis in MKN-45 cells, accompanied by increased Cyt-C, decreased BCL-2, and decreased mitochondrial membrane potential and mitochondrial damage. Further research showed that quercetin targeted EGFR, blocked the ERK signaling pathway, and downregulated PTGS2. In the in vivo experiment, quercetin treatment resulted in reduced tumor volume, decreased Ki67 and BCL-2 expression in tumor tissue, increased caspase 3 and BAX levels, and induced mitochondrial damage.

The late stage of embryo development is a crucial period of metabolic changes, with rapid organ development requiring a substantial supply of nutrients. During this phase, maternal nutritional levels play a vital role in the growth, development, and metabolism of the offspring. In this study, we added 2 doses of β-carotene (βc) (120 mg/kg and 240 mg/kg) to the daily diet of Hailan Brown laying hens to investigate the impact of maternal nutritional enrichment on embryo development. Maternal nutrition supplementation significantly increased the expression of chicken embryo liver index, growth hormone (GH), insulin-like growth factor-1 (IGF-1), and hepatocyte growth factor (HGF) in serum. At the same time, the expression of GH/growth hormone receptor (GHR), IGF-1 mRNA, and Proliferating Cell Nuclear Antigen (PCNA) protein in the liver was upregulated, indicating that maternal nutrition intervention may promote chicken embryo liver development through the GH-IGF-1 axis. Transcriptome sequencing results showed that differential genes in liver after maternal nutritional supplementation with β-carotene were enriched in pathways related to cell proliferation and metabolism. Consequently, we postulated that maternal β-carotene supplementation might operate via the GH-IGF-1 axis to regulate the expression of genes involved in growth and development, thereby promoting liver development. These results contribute to formulating more effective poultry feeding strategies to promote offspring growth and development.

Objective: To explore the role and molecular mechanism of cancer-associated fibroblasts (CAFs) in the tumor microenvironment of gastric cancer (GC). Methods: The expression of CAFs in GC patients was first assessed for abundance, and survival analysis was performed. Subsequently, The Cancer Genome Atlas (TCGA) data were used for differential analysis, survival analysis, and EPIC analysis, while single-cell data (GSE183904) were downloaded for differential analysis of CAFs. Clinical data pooling, univariate and multivariate Cox analysis, and immunofluorescence were carried out on clinical GC tissue samples to explore RCN3 expression within patient CAFs. Western blot and quantitative polymerase chain reaction (qPCR) were used to detect the expression of RCN3. The relationship between RCN3, PCSK6, and STAT1 was explored by chromatin immunoprecipitation (CHIP) experiments, and the effects of the genes on macrophage polarization were detected by detecting biomarkers of biological M1/M2. Results: CAFs in GC were found to be significantly higher compared to the normal group. Revealing the results of TCGA differential analysis, it was observed that GC exhibited a substantial upregulation in the expression levels of RCN3. The clinical statistics indicate a positive correlation between an elevated level of RCN3 expression and the T-stage classification of tumor size. In addition, RCN3 was found to have a significant impact on the overall survival of patients with gastric cancer, acting as an independent prognostic indicator. Analysis of single-cell data showed high expression of PCSK6 in macrophages, and immunofluorescence staining of samples from GC patients showed increased expression of PCSK6 on the cell membranes of macrophages in GC tissues. The subsequent cellular experiments confirmed RCN3 protein can regulate the expression of PCSK6, and PCSK6 regulates macrophage polarization through STAT1. Conclusions: CAFs regulate macrophage polarization through the RCN3/PCSK6/STAT1 pathway in GC.