Objective The pathogenesis of type 2 diabetes mellitus (T2DM) is intrinsically linked to the suboptimal secretion of glucagon-like peptide-1 (GLP-1). While postbiotics have emerged as a compelling therapeutic modality for metabolic modulation, the precise bioactive effectors and their underlying mechanisms remain largely elusive. This study aimed to evaluate the effect of Bifidobacterium breve i1088 (B breve i1088) lysate on GLP-1 secretion and identify its key active molecules. Methods The GLP-1 secretory activity of i1088 lysate was assessed in human enteroendocrine NCI-H716 cells. Subsequently, the metabolic benefits of the lysate were validated in a T2DM mouse model. Untargeted metabolomics was employed to profile the bioactive substances of the lysate. Results In vitro, B breve i1088 lysate exhibited a robust, dose-dependent capacity to stimulate GLP-1 secretion. In vivo, high-dose lysate (HL) not only recapitulated the glucose and lipid-lowering and anti-inflammatory efficacy of live bacteria (LB), but also demonstrated superior stability. Mechanistically, HL treatment profoundly reprogrammed the intestinal incretin axis by upregulating preproglucagon (GCG) and proprotein convertase 1 (PC1), thereby enhancing endogenous GLP-1 synthesis and preserving pancreatic islet function. Crucially, we identified 5-hydroxyindoleacetic acid (5-HIAA) as the pivotal bioactive signature within the B breve i1088 lysate. Cellular assays confirmed that 5-HIAA directly triggered GLP-1 release and upregulated the mRNA expression of both the GCG and PC1 genes, serving as a key molecular messenger in the B breve i1088-mediated metabolic recovery. Conclusion HL enhances GLP-1 synthesis via the upregulation of the GCG/PC1 pathway and alleviates T2DM, with 5-HIAA serving as a key functional metabolite in this postbiotic-mediated response. The potent efficacy of the B. breve i1088 lysate highlights its potential as an innovative postbiotic therapy for T2DM management.

The rapid destabilization of fat in direct ultra-high-temperature (UHT) milk during storage is mainly caused by plasmin through the cleavage of interfacial proteins. In this study, novel fat globules were developed by modulating the interfacial proteins (milk fat globule membrane material homogenized with large fat globules) to resist plasmin hydrolysis and improve fat stability. Compared to simulated homogenized milk (milk proteins homogenized with large fat globules), the novel fat globules contained more proteins at the interface and showed a greater stability. Proteomics revealed that the protein types at the interface of novel fat globules were predominantly membrane proteins, while the interface of simulated homogenized milk was mainly composed of milk proteins. The relative abundance ratio of major MFGM proteins to casein in the novel fat globules was approximately 30 times higher than in simulated homogenized milk. After plasmin hydrolysis, the novel fat globules showed minimal changes in stability, with only small amounts of XO and casein being hydrolyzed. In contrast, the fat globules in the simulated homogenized milk underwent coalescence, and a large amount of casein was hydrolyzed at the interface. Furthermore, the novel fat globules were more stable than commercial direct UHT milk, both before and after plasmin intervention.

Following the aging of the population, cognitive impairment is becoming a major public health issue. Research in recent years has highlighted the potential of docosahexaenoic acid (DHA), folic acid (FA), and phosphatidylserine (PS) to improve synaptic plasticity and reduce inflammatory response. However, whether a combined intervention is more effective than using these supplements alone remains unclear. To investigate this, we conducted an experiment on D-galactose-induced aging mice and found that the combining DHA, FA, and PS (DFP) improved spatial learning memory. Moreover, the DFP treatment restored cognitive performance by regulating antioxidant levels, inhibiting microglia activation, and reducing inflammation. The treatment also increased the expression of neurotrophic factors and synaptic-related proteins in the hippocampal region of the brain. Our findings suggest that the supplementation of DPF could be more beneficial than using DHA alone. This combined intervention could improve neurotrophic factor expression and decrease neuroinflammation in the brain of mice, thus reducing neuron damage and enhancing synaptic plasticity. In summary, our results indicate that the combined supplementation of DFP may improve cognition, and may be more effective than supplement with DHA alone.

The proliferation of small intestinal epithelial cells is fundamental to intestinal development and barrier function maintenance in infants and young children. To investigate the effects and mechanisms of u03C9-3 polyunsaturated fatty acids (docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA)), commonly used in infant formula, on this process, this study employed the rat intestinal epithelial cell line 6 (IEC-6) as a model. After DHA and EPA treatment at different combinations of concentration (1, 10, 50, 100 u03BCmol/L) and duration 12, 24, 36, 48 h, cell viability was assessed using the cell counting kit-8 and 5-ethynyl-2u2019-deoxyuridine assays, and the mRNA and protein expression of key molecules involved in the Wnt/u03B2-catenin signaling pathways were analyzed via real-time quantitative polymerase chain reaction and Western blot. The results showed that both DHA and EPA significantly promoted the proliferation of IEC-6 cells, with optimal concentrations of 100 and 10 u03BCmol/L, respectively, and an optimal treatment duration of 24 h. Specifically, treatment with 100 u03BCmol/L DHA and 10 u03BCmol/L EPA for 24 h increased cell viability by nearly 4- and 3.5-fold compared to the control group (P u0026lt; 0.001), respectively. Both DHA and EPA activated the Wnt/u03B2-catenin signaling pathway, significantly upregulating the mRNA and protein expression of cell proliferation-related genes such as cyclin D1 and u03B2-catenin (P u0026lt; 0.05, P u0026lt; 0.01, or P u0026lt; 0.001). These findings indicate that DHA and EPA enhance intestinal epithelial cell proliferation by activating the Wnt signaling pathway.

To develop an ultrafiltration process for separating α-lactalbumin (α-La) and achieve high-value utilization of by-products, this study employed a small-scale tangential flow ultrafiltration equipment to optimize the process parameters for enriching α-La from whey protein concentrate. Additionally, the study investigated the heat treatment modification of the by-product after α-La separation. The results showed that the optimal conditions for α-La separation were as follows: Feed pH of 6.5, ultrafiltration membrane molecular weight cutoff of 100 kDa, volume concentration factor of 4, operating pressure of 0.3 MPa, and 5 diafiltration cycles. Under these conditions, the ratio of α-La to β-lactoglobulin (β-Lg) in the permeate increased to over 9, a 23-fold improvement compared to the whey protein concentrate. The foaming properties of the β-Lg-rich retentate, remaining after α-La separation, were optimal under heat treatment at 60 ℃ for 10 minutes, with foam ability and foam stability reaching 18.1% and 69.9%, respectively. In contrast, the emulsifying properties were optimal at 80 ℃ for 10 minutes, with emulsifying activity index and emulsifying stability index reaching 77.0 m2/g and 82.8%, respectively. Overall, this study established an optimal membrane separation process for enriching α-La and enhanced the functional properties of the by-product retentate through heat treatment, providing a valuable technical solution for the industrial production of α-La and the comprehensive utilization of by-products.

Microfluidics technology emerges as a transformative solution in drug development paradigms. The demand for microfluidics technologies for pharmaceutical science accelerated to the development of Pharm-Lab on a Chip. This review aims to inform how the Pharm-Lab on a Chip facilitates pharmaceutical analysis. First, we survey innovative microfluidics-driven methodologies for drug synthesis, separation and analytical characterization, highlighting technological breakthroughs in molecular isolation and quantification. Subsequently, we highlight the recent pioneering work in developing microfluidic-based biomimetic organ models, and micro-physiological environment models. Notably, we present groundbreaking pharmacological and toxicological evaluation platforms that enable high-fidelity assessment of drug efficacy profiles and safety parameters through precise microenvironmental control. In the end, we discuss future challenges that need to be addressed before microfluidics can be widely used for drug analysis. We believe that this review could provide a systematic and in-depth understanding of microfluidics-based pharmaceutical analysis.

Lactoferrin (LF) is a multifunctional iron-binding glycoprotein that possesses a variety of functions, such as antibacterial, anti-inflammatory, antitumor, and intestinal health promotion. However, its application was restricted by its instability and poor gelation property. Recently, an increasing number of studies have focused on the synergistic effects of LF to broaden its application in the fields of functional food, nutritional supplements, and health care. LF can form complexes with other substances or be modified to enhance its stability and functionality. Additionally, the charge characteristics and cage-like structure of LF enable it to act as a carrier to encapsulate and protect functional ingredients. Furthermore, due to its broad range of receptors, LF can serve as a targeted delivery material for the precise delivery of functional ingredients. New insights into future research trends were also present, which are expected to further expand the utility of LF.

The role of CD8+ T cells in the pathogenesis of ulcerative colitis (UC) remains unclear. Similarly, the posttranscriptional regulation of the highly heterogenic CD8+ T cell populations and their effector function in IBD also remains poorly understood. Here, we find that miR-29a and -29b (miR-29a/b) regulate T cell fate, and their expression is higher near damaged colon tissue in patients with IBD compared to controls. In mice, we find that miR-29a/b suppresses the differentiation of CD8+ T cells and the secretion of pro-inflammatory and chemotactic factors during severe colitis by inhibiting transcriptional pathways, including those involving the T cell receptor and JAK-STAT signaling. Furthermore, we identify Ifng, an inflammatory factor that drives immune response and the reshaping of CD8+ T cell fate, as a potential target of the miRNAs. Finally, we show that delivery of miR-29 mimics to the colon of mice is sufficient to alleviate DSS-induced inflammation. Together, these data show that miR-29 plays an important role in suppressing T cell overactivation during inflammatory diseases.

Zhaiji millet vinegar (ZJMV) is a traditional grain-fermented condiment whose quality improvement is associated with aging time. This study revealed the evolution of physicochemical properties and whole-component of ZJMVs aging 2 to 10 years. Results demonstrated pH, chroma, and the content of total acid, reducing sugar, and amino acid nitrogen fluctuate dynamically, while antioxidant activities were significantly correlated with the content of total flavonoids, total phenols, tetramethylpyrazine, amino acids, and organic acids (p < 0.05). GC-IMS and metabolomics identified 47 volatile organic compounds (VOCs) and 6530 metabolites, with 6 differential VOCs and 19 differential metabolites (p < 0.05, variable importance in projection (VIP) > 1, fold change (FC) < 0.5 or > 2) being shared aging markers. KEGG pathway analysis indicated amino acid metabolism enriched the most differential metabolites, whereas biosynthesis of various alkaloids was critical. Our findings provide novel mechanistic insights into the quality development of ZJMV during aging.

Human milk oligosaccharides (HMOs) have been shown to have multiple benefits, including enhance intestinal barrier, antioxidant and anti-inflammatory properties, and therefore have the potential to improve the aging intestinal barrier. Aging can lead to damage to the intestinal barrier and function, which can affect human health. However, the effectiveness and mechanism of HMOs with different structures in improving the aging intestinal barrier are still unclear. Here, we used honey bee to examine the effects of HMOs on physiology, intestinal barrier, and gut microbiota. The results showed that HMOs reduced mortality rate, improved the intestinal barrier, and increased the mucus layer thickness in aging honey bee. We also found that 2'-FL and LNnT boosted the proportion of Bifidobacteria in the bee gut microbiota, while 3'-SL elevated the proportion of Gilliamella. HMOs increased the content of leucine in intestinal metabolites and reduced oxidative stress levels through tryptophan metabolism. In conclusion, our study showed that HMOs can improve the aging gut barrier via altering gut microbiota. The improvement effect of 2'-FL is better than that of 3'-SL and LNnT. The study provided a theoretical basis for the development of new functional ingredients to improve the aging gut.

Depression is the main cause of the global mental health burden and was recently reported to be correlated with constipation[1,2].Scattered reports have demonstrated that the association between mood disorders and gastrointestinal disturbances may be related to cross-talk between the gut and the central nervous system,referred to as the gut-brain axis[3].

Milk-derived small extracellular vesicles (M-sEVs) represent a key bioactive component of breast milk, demonstrating multifaceted benefits for intestinal health, including preservation of intestinal barrier integrity, modulation of gut microbiota composition, immunoregulatory functions, and amelioration of colitis. However, their regulatory roles in early-life intestinal and immune development remain incompletely elucidated. Here, M-sEVs derived from human breast milk significantly enhanced body weight, body length, and small intestinal length in suckling mice. Further analysis revealed that M-sEVs potently stimulated intestinal epithelial regenerative capacity and increased the population of secretory lineage epithelial cells (including goblet cells, Paneth cells, and enteroendocrine cells). In addition, M-sEVs administration regulated immune programming in the intestinal lamina propria, enhanced the differentiation of IgA+ plasma cells, upregulated the proportions of CD4+ and CD8+ T cells, and exerted regulatory effects on overall immune development. Finally, we identified that M-sEVs upregulated genes and pathways associated with immune regulation, proliferation and differentiation, as well as digestion and absorption, thereby playing a pivotal role in intestinal maturation during infancy. Overall, this study provides novel insights into the potential role of M-sEVs in modulating intestinal epithelial and immune development during infancy, thereby establishing a scientific foundation for developing early-life nutritional intervention strategies.

Beta-casein (β-CN) and whey protein are highly promising ingredients for infant milk formula (IMF), and their addition to IMF can further bridge the nutritional gap between IMF and human milk. At present, there are few studies on pilot-scale co-enrichment of β-CN and whey protein, which is crucial for industrial-scale production. This study investigated the optimization of pilot-scale process parameters for the co-enrichment of β-CN and whey protein through cold microfiltration. A pilot-scale 3-stage ceramic membrane filtration system was used, and two different membrane pore sizes, 0.14 μm and 0.2 μm, were evaluated. The results demonstrated that the 0.14 μm ceramic membrane exhibited higher filtration efficiency than the 0.2 μm membrane. Additionally, the effects of transmembrane pressure (TMP, 50–140 kPa), volume concentration factor (VCF, 2–3.5), and filtration temperature (10–25 °C) on process performance and protein-enrichment were assessed using the 0.14 μm membrane. The results indicated that the enrichment rate increased with the rise in TMP, VCF and temperature. Filtration efficiency also increased with TMP and temperature; however, it initially increased and then decreased as the VCF increased. The optimal TMP, VCF, and temperature for high-efficiency enrichment of β-CN and whey protein were 140 kPa, 3, and 15 °C, respectively. These findings will provide valuable guidance for the future industrial-scale production of β-CN and whey protein.

Fatty acids (FAs) have various functions on cell regulation considering their abundant types and metabolic pathways. In addition, the relation between FA and other nutritional metabolism makes their functions more complex. As the first place for diet-derived FA metabolism, intestine is significantly influenced despite lack of clear conclusions due to the inconsistent findings. In this review, we discuss the regulation of fatty acid metabolism on the fate of intestinal stem cells in homeostasis and disorders, and also focus on the intestinal tumor development and treatment from the aspect of gut microbiota-epithelium-immune interaction. We summarize that the balances between FA oxidation and glycolysis, between oxidative phosphorylation and ketogenesis, between catabolism and anabolism, and the specific roles of individual FA types determine the diverse effects of intestinal FA metabolism in different cases. We hope this will inspire further dissection and suggest precise dietary/metabolic intervention for different demands related to intestinal health.

BACKGROUND:Food-derived extracellular vesicles (FEVs) from plants, milk and probiotics are emerging as bioactive nanocarriers linking nutrition and metabolism. Their natural stability provides unique advantages for gastrointestinal therapeutics. Inflammatory bowel disease (IBD) is a refractory intestinal disease, and the current research shows that FEVs have potential application value in improving IBD. Scope and Approach: This review systematically summarizes the current understanding of FEVs from three major sources-plant-derived, milk-derived, and bacteria-derived EVs, encompassing their biogenesis, structural characteristics, application characteristics and mechanism of action, with emphasis on their role in IBD intervention. And put forward new strategies and prospects for FEVs' intervention and application in IBD, providing a comprehensive perspective for their role in nutrition intervention. KEY FINDINGS AND CONCLUSIONS:FEVs from distinct sources exhibit characteristic structural and functional profiles that enable source-specific intervention strategies for IBD. These natural nanovesicles demonstrate multi-modal therapeutic effects by modulating gut microbiota composition, enhancing intestinal barrier integrity, and fine-tuning immune responses. Their role as nutritional modulators and drug delivery vehicles holds significant clinical promise for IBD management. However, translational challenges persist, including mechanism uncertainties, suboptimal targeting efficiency, unverified long-term safety, and limited production yields. Future studies should prioritize bridging these knowledge gaps to facilitate FEV-based clinical translation.

β-casein (β-CN) is the predominant casein fraction in breast milk, while current infant milk formula (IMF) contains substantially lower β-CN levels than breast milk. The impact of β-CN fortification on neonatal digestive characteristic and bioactive peptide release remains an understudied area in vivo. This study investigated the effect of β-CN fortification in milk protein on digestion properties and release of bioactive peptides using a suckling rat pup model. Rat pups were, respectively, gavaged with two milk protein solutions: one with ordinary β-CN content (OBCN) and the other with fortified β-CN content (FBCN). The gastric emptying rate, proteolytic efficiency, and peptidomic profiles of intestinal digesta were evaluated. Results indicated that the FBCN group exhibited accelerated gastric emptying into the intestinal phase and enhanced proteolytic efficiency compared to OBCN group. Furthermore, the FBCN group generated greater peptide diversity in the small intestine, with significantly elevated abundance of bioactive peptide candidates exhibiting broader functional spectra. These findings provide additional evidence for the health effects of β-CN fortification in IMF.