Yuncai Xiao

Both carboxymethyl chitosan (CMCS) and Mn²⁺ can stimulate the cGAS-STING pathway to promote the production of immune factors such as type I interferons (I IFNs), thereby regulating the body’s immune response. In this study, a sustained-release Mn²⁺-loaded CMCS-Mn hydrogel was designed, and the synergistic effects of CMCS and Mn²⁺ on innate immunity, as well as factors influencing the immune response, were investigated. The optimal gel formulation was obtained with a CMCS concentration of 1 mg/mL and a MnCl₂ concentration of 0.02 mol/L, yielding a gel with a particle size of 472.16 nm, a polydispersity index of 0.35, a Mn²⁺ content of 1.488 mg/g, and a specific surface area of 4.74 m²/g. The hydrogel forms mainly via coordination interactions between Mn²⁺ and the amino and carboxyl groups on CMCS molecular chains and exhibits pH-dependent Mn²⁺ release behavior. Cellular assays demonstrated that the combination of 600 μg/mL CMCS-Mn²⁺ hydrogel and 5 μg/mL free Mn²⁺ produced the strongest IFN-β induction. The hydrogel exhibits synergistic immunomodulatory effects of both CMCS and Mn²⁺; while the amino groups are critical for the immunological activity of CMCS, Mn²⁺ plays a more prominent role. Partial substitution of Mn²⁺ by Ca²⁺ can loosen the cross-linked network and further modulate Mn²⁺ dissociation from the hydrogel.

Weaning impairs intestinal function and growth performance in piglets. This study evaluated a fermented herbal formulation (FHF) composed of five bioactive herbal ingredients—Radix isatidis, Folium isatidis, Radix scutellariae, Fructus forsythiae, and Radix glycyrrhizae—fermented with Enterococcus faecium and Saccharomyces cerevisiae and characterized by flavonoids, phenolic acids, and hydroxylated fatty acids, using the porcine intestinal epithelial cell line (IPEC-J2) and weaned piglets. In vitro, IPEC-J2 cells were pretreated with FHF extract (100–1000 μg/mL) for 3 h prior to lipopolysaccharide (LPS) challenge. In vivo, 72 piglets were weaned at 32 days of age and, after a 3-day post-weaning adaptation period, entered a 35-day feeding trial. The piglets were then randomly assigned to three treatment groups: control (basal diet), A1 (basal diet + 0.4% FHF), and A2 (basal diet + 0.6% FHF during days 1–18, followed by 0.3% FHF during days 19–35). FHF dose-dependently alleviated the LPS-induced decrease in cell viability and suppressed IL-6, IL-8, IL-1β, and TNF-α expression. In piglets, the A2 group showed higher final body weight, average daily gain (ADG), and average daily feed intake (ADFI), lower feed conversion ratio (FCR), and a lower diarrhea rate than the control group. FHF also improved intestinal morphology, reduced serum TNF-α and diamine oxidase (DAO) levels, increased jejunal tight junction protein expression, enriched Limosilactobacillus and Lactobacillus, and elevated acetic and butyric acids. FHF improved intestinal health and growth performance in weaned piglets, with the A2 group showing the best overall efficacy.

Gut microbiota plays a pivotal role in host immunity and overall health. Enterococcus faecium HDRsEf1 (Ef1), a novel potential probiotic, has been reported to enhance immune defense against pathogenic invasion; however, its effects on inflammation and gut microbiota remain poorly understood. This study aimed to evaluate the impact of Ef1 on host health, gut microbiota, and fecal metabolite profiles using DSS-induced colitis in mice and nursery pig models. In mice, Ef1 effectively mitigated growth retardation and organ damage caused by DSS-induced inflammation. In nursery pigs, Ef1 administration reduced markers of inflammation and oxidative stress, including IL-1β, IL-6, TNF-α, and MDA, while increasing the anti-inflammatory cytokine IL-10 and the antioxidant enzyme SOD. Furthermore, Ef1 modulated gut microbiota composition, promoting beneficial genera such as Ruminococcus and Acetobacter , while suppressing potentially harmful genera like Acinetobacter . Metabolomic analysis revealed that Ef1 enriched anti-inflammatory metabolites in the gut. Collectively, these findings indicate that Ef1 can beneficially modulate gut microbiota and metabolite profiles, thereby enhancing host health.

Introduction: Lung cancer is a highly heterogeneous disease, and the tumor microenvironment (TME) characteristics are closely related to disease progression and treatment outcomes. We elucidated the cell type-specific transcriptome landscape of cancer cells and the effect of the TME on lung adenocarcinoma (LUAD). Methods: Single-cell RNA sequencing of the LUAD tissues and matched adjacent normal tissues of three patients in the early-stage via histopathological and immunohistochemical confirmation. Results: The results revealed the landscape of 68,579 cells in LUAD microenvironment, and highly heterogeneous AT2 cells are crucial source of lung epithelial cell carcinogenesis. Genes KRT81 , SPP1 , PCDH7 , SLC2A1 , and TET1 were significantly upregulated in tumor tissues and associated with poor prognosis and survival, providing insights for exploring lung cancer biomarkers in future studies. Trajectory analysis identified ERBB4 , SEMA4A , GCNT2 and SOX4 as key factors in AT2 cells that may promote cell proliferation, migration, or epithelial-mesenchymal transition (EMT) during the progression of lung adenocarcinoma (LUAD). We elucidated the transcription factor–target gene regulatory network involving NKX2-1 and TEAD1 in malignant tumor cells derived from AT2 cells. The findings indicated that malignant AT2 cells regulate communication between epithelial and immune cells in the TME by predicted FN1-CD44 and CADM1-CADM1 ligand-receptor interactions, which ultimately suppresses the host immune response. Discussion: This comprehensive single-cell analysis increases our understanding of AT2 cells molecular and dynamics of metastatic lung cancer. In summary, single-cell RNA profiling of LUAD offers valuable prognostic insights based on AT2 cell types and identifies potential biomarkers for therapeutic responses, aiding the future development of LUAD treatment strategies.

Background: Enteropathogenic Escherichia coli (EPEC) disrupts intestinal barrier integrity by adhering to epithelial cells, leading to diarrhea, impaired nutrient absorption, oxidative stress, and intestinal inflammation in young animals. This study aimed to isolate and characterize a neutral exopolysaccharide (EPS-T1) from Enterococcus faecium HDRsEf1, evaluate its functional activities in vitro, and assess its protective effects against EPEC-induced enteritis in vivo. Results: EPS-T1, with a molecular weight of 81.21 ± 1.28 kDa, was mainly composed of glucose, galactose, rhamnose, and mannose, and exhibited a porous, sheet structure with relatively high thermal stability. In vitro, EPS-T1 (200 μg/mL) significantly inhibited EPEC growth and biofilm formation, reduced bacterial adhesion to intestinal epithelial cells, and exhibited broad-spectrum free radical scavenging activity. In vivo, EPS-T1 treatment alleviated EPEC-induced weight loss and intestinal tissue damage, reduced the intestinal EPEC load, downregulated pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6), upregulated the anti-inflammatory cytokine IL-10, and improved serum antioxidant indices (T-AOC, SOD, and GSH-PX) while decreasing MDA levels. Conclusions: These results demonstrate that EPS-T1 derived from Enterococcus effectively mitigates EPEC-induced intestinal inflammation and oxidative stress, highlighting its potential as an immunobiotic functional candidate.

This study aimed to identify the heat-resistant bioactive components of Enterococcus faecium HDRsEf1 (HDRsEf1) and investigate their beneficial mechanism. Heat-treated culture supernatants of HDRsEf1 significantly suppressed CXCL-1 expression in LPS-stimulated MODE-K cells (p < 0.001), indicating the presence of heat-resistant anti-inflammatory components. Crude protein (P-Ef1) and crude expolysaccharide (EPS-Ef1) were isolated from an HDRsEf1 culture supernatant using ammonium sulfate and ethanal precipitation. Critically, only crude EPS-Ef1 retained an anti-inflammatory effect after heat treatment, while crude P-Ef1 lost this activity. Further investigation revealed that crude EPS-Ef1 (25 μg/mL) promoted MODE-K cell proliferation via EdU assays (p < 0.001), potentially through an upregulation of PCNA mRNA expression (p < 0.001). Animal studies demonstrated that an oral administration of crude EPS-Ef1 (4 mg/kg bw, 14 days) significantly increased body weight gain and jejunal crypt depth (p < 0.05) while reducing intestinal CXCL-1 mRNA levels (p < 0.001). These in vivo findings are consistent with in vitro observations. A structural analysis using HPAEC and SEC-MALLS-RI characterized crude EPS-Ef1 as a heteropolysaccharide (Mw 80.3 kDa) with a near-spherical conformation (slope 0.13) composed of mannose, glucose, glucuronic acid, and galactose (5.4:4.4:1.2:1). In summary, this study identifies crude EPS-Ef1 as the heat-resistant postbiotic component. Crude EPS-Ef1 possesses the dual effects of suppressing intestinal inflammation and promoting intestinal epithelial cell proliferation, which provides a theoretical foundation for a crude EPS-Ef1-based postbiotic.