Agrocybe aegerita (A. aegerita), a globally cultivated mushroom, is increasingly recognized as a functional food owing to its rich profile of bioactive compounds and diverse health benefits. This review evaluates its composition and bioactivities. Predominant macronutrients are carbohydrates (84.50 g/100 g DW) and proteins (6.68 g/100 g DW), alongside essential trace elements and unsaturated fatty acids (PUFAs, 78.60

Chitooligosaccharides (COSs) are hydrolysates of chitosan polymers. COSs have many biological functions, especially in immune activation. Gut microbiota is closely related to the decomposition of natural glycans in the human intestine and plays a pivotal role in regulating the immune system. But the role and mechanism of intestinal microbiota in mediating the effects of COSs on the immune system remain unclear. In this study, BALB/c mice were gavaged with 100 μL of COSs at a concentration of 0.1 g/mL for 60 days, and the intestinal microbiota and immune cells of the mouse spleen were detected. Administration of COSs altered the gut microbiota in mice, especially promoting the abundance of Lactobacillus spp., which was consistent with the observation in the fecal sample from volunteers who received COSs. The proportion of CD3+ T cells in the spleen was improved after intragastric administration of COSs. COSs also promoted the growth of Lactobacillus spp., such as L. brevis and L. gasseri, in a dose-dependent manner, and the supernatant from co-cultured medium promoted the proliferation of Jurkat cells (human T cell leukemia). The expression of β-1,4-glycosidase significantly contributed to the utilization of COSs by these Lactobacillus strains. In addition, we utilize protein sequencing and structure analysis to examine the differences of β-1,4-glycosidase among Lactobacillus strains. Taken together, our results suggested that COSs can promote T cell proliferation by supporting the growth of intestinal Lactobacillus spp. in mice, which further revealed the immuno-regulation mechanism of COSs in the human body and its application prospects in health promotion.

Selenium-enriched Pleurotus ostreatus is a promising source of selenopolysaccharides (Se-PPs), but efficient extraction and solvent recovery remain challenging. In this study, an integrated strategy combining a pH-switchable linalool/octanoic acid deep eutectic solvent (DES) with high-speed shearing and ultrasonication was developed. Solvent formulation and extraction conditions were optimized by single-factor experiments and response surface methodology. Hot-water extraction with matched high-speed shearing and ultrasonication (HSU-HWE) served as the comparator. Under optimized conditions, the Se-PPs yield was 146.94 ± 1.28 mg/g with HSU-DES and 74.80 ± 1.60 mg/g with HSU-HWE. On the seventh use, DES-5 retained 97.65% of its initial extraction capacity. FT-IR, D2O-based 1H NMR, and 2D ROESY analyses supported local intermolecular association between linalool and octanoic acid. DFT calculations using glucose as a representative saccharide unit identified plausible non-covalent contacts among the DES components and glucose. These results are consistent with local molecular interactions that may contribute to DES formation and polysaccharide recovery. HSU-DES-Se-PPs contained less water-extractable Se(IV) + Se(VI) and had a higher calculated organic selenium fraction than HSU-HWE-Se-PPs. They also had lower IC50 values for DPPH, ABTS•+, and hydroxyl-radical scavenging (1.04, 3.42, and 1.00 mg/mL, respectively). ComplexGAPI and an AGREE score of 0.72 indicated a comparatively favorable laboratory-scale greenness profile for HSU-DES. Overall, the results support HSU-DES as a reusable approach to Se-PPs recovery under the tested laboratory conditions.

Most broiler heat stress studies focus on the exposure phase, whereas the first hours after heat withdrawal remain understudied. We tested whether a flavonoid extract from Bupleurum aerial parts (BAPF), administered at recovery onset, could reduce residual liver and lung injury in yellow-feathered broilers exposed to 35°C for 6 h and then cooled to 24°C over the next 6 h. The extract improved liver and lung histology, lowered serum injury markers and circulating interleukin-1β and interleukin-18, and improved hepatic redox indices, with the most consistent responses at 46 to 68 mg/kg body weight per day. Exploratory composite analysis also prioritized the 46 to 68 mg/kg body weight per day range. In a separate heat and rewarming model using the chicken fibroblast cell line DF-1, the extract added at recovery onset lowered reactive oxygen species, increased nuclear abundance of nuclear factor erythroid 2-related factor 2, and reduced immunoblot signals for NLR family pyrin domain containing 3, cleaved caspase-1, cleaved caspase-3, and cleaved gasdermin E. An ML385 pretreatment arm attenuated several of these changes, supporting involvement of this pathway. Overall, the findings suggest that early recovery after heat exposure may provide a post-heat intervention window and warrant further evaluation of BAPF initiated at recovery onset to limit residual organ injury in yellow-feathered broilers.

Despite significant progress in understanding chitin oligosaccharides as plant immune elicitors, studies on their potential to induce insect resistance remain limited. In this study, we demonstrate that chitin oligosaccharides induce potent and broad-spectrum insect resistance in Brassica napus. NACOS treatment significantly reduced damage from both chewing insects (Plutella xylostella) and piercing-sucking insects (Myzus persicae). Multiomics analyses revealed that NACOS activated defense-related pathways, including peroxisome, phenylpropanoid biosynthesis, and glutathione metabolism, as evidenced by enhanced activities of superoxide dismutase, peroxidase, and polyphenol oxidase, as well as the accumulation of defense-related metabolites such as GABA, tannins, and oxalic acid. While inducing robust defense responses, NACOS also upregulated photosynthesis-related pathways, leading to improved photosynthetic rates and markedly promoting plant growth and yield. Our study uncovers a dual-role mechanism of NACOS in balancing defense and growth, providing a theoretical basis and practical strategies for developing oligosaccharide-based biopesticides.

Licorice, commonly named as ''sweet grass'', is a perennial herbaceous plant of the Leguminosae family, and licorice roots possess functions related to clearing heat and detoxifying toxins, tonifying qi, and harmonizing the properties of other herbs in traditional Chinese medicine. Among its bioactive ingredients, licorice polysaccharides (LPs) have considerable health benefits. Hot water extraction, acid/alkali extraction, and ultrasound-assisted extraction are frequently used for extracting LPs. LPs are purified from different fractions, while only a few of them are well characterized. LPs exhibit anti-tumor, anti-colitis, hepatoprotective, hypoglycemic, antiviral and immunomodulatory activities. In recent years, multiple studies have been conducted on deciphering structural features and pharmacological activities of LPs extracted by different methods. Nevertheless, the lack of clarity regarding the structure-pharmacology correlations of LPs represents a significant barrier to understanding their therapeutic mechanisms. Further research on higher-order structure and health-related molecular mechanisms of LPs is crucial for their high value utilization. This review comprehensively summarizes the preparation methods, structural characteristics, pharmacological activities and related to mechanisms of LPs. In addition, this review highlights their promising applications in medicine, food industry and livestock farming industry. Furthermore, this paper deliberates on future perspectives of LPs, aiming to offer novel insights and trajectory for the future development of LPs.

Alginate oligosaccharides (AOS) are degradation products of alginate, exhibiting antioxidant, antibacterial, and anti-inflammatory activities. This study aimed to investigate the protective effect and molecular mechanism of AOS against lipopolysaccharide (LPS)-induced sepsis-associated liver injury (SALI) through in vivo and in vitro experiments. In vivo investigation indicated that AOS significantly mitigated LPS-induced hepatic injury in Balb/c mice by suppressing the overexpression of pro-inflammatory cytokines, including interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β). Transcriptomic analysis confirmed that AOS regulated the mitogen-activated protein kinase (MAPK) and nuclear factor kappa-B (NF-κB) signaling pathways. In vitro assays demonstrated that AOS significantly reduced the production of nitric oxide (NO) and the aforementioned pro-inflammatory factor levels in RAW264.7 cells. Moreover, AOS treatment inhibited the phosphorylation of p38, Erk, and p65, suppressed the nuclear translocation of p65, and consequently attenuated LPS-induced activation of both the MAPK and NF-κB signaling pathways. Mechanistic studies further indicated that AOS suppressed inflammation by activating autophagy, and this protective effect was reversed by an autophagy inhibitor. Collectively, AOS alleviated LPS-induced SALI via its anti-inflammatory effects and activation of autophagy. These findings provide an experimental foundation for adjuvant nutritional strategies in SALI and support the potential of AOS as a hepatoprotective functional food ingredient.

BACKGROUND:The increasing challenges of pesticide resistance and environmental degradation in modern agriculture require sustainable pest management strategies. This study investigates the efficacy of a specific type of hetero-chitooligosaccharide called partially deacetylated chitooligosaccharide enriched in d-glucosamine (DACOS; 85.6% deacetylation, an average molecular mass of 1092.52 Da), as a plant defense elicitor against the two-spotted spider mite, Tetranychus urticae Koch. RESULTS:In planta bioassays revealed suppression of mite performance on DACOS-treated leaves of kidney bean seedlings. Local effects were most pronounced at 80 mg L-1 (ppm), which reduced mite survival by 30% and cumulative fecundity by 41% after 6 days. Systemic effects were strongest at 160 ppm, indicating distinct concentration requirements for local versus systemic defense induction. Behavioral choice assays revealed that DACOS-treated leaves induced 73% mite repellency. No direct acaricidal effects were observed, which confirms the role of DACOS as a plant defense elicitor. Metabolic profiling revealed enriched pathways (e.g., monoterpenoid biosynthesis and α-linolenic acid metabolism) that produce volatile organic compounds with repellent properties. CONCLUSION:These findings suggest that DACOS is an environmentally friendly and non-biotic induced resistance stimulus that can enhance plant defense, bypass pesticide resistance, and promote the sustainability of crop production. © 2026 Society of Chemical Industry.

Polysaccharides are natural polymers that are widely found in medicinal plants. Structurally, they are complex molecules composed of long chains of monosaccharide units linked by glycosidic bonds. Modern pharmacological research shows that the bioactivity of polysaccharides is closely related to their monosaccharide composition. This review summarises the monosaccharide composition of 210 polysaccharides from 72 medicinal plants. They were classified into 10 types through principal component analysis (glucans; homogalacturonan; galactans; arabinogalactans; mannans; glucomannans; arabinans; xylans; fructans; rhamnogalacturonan-I). The relationship between monosaccharide composition and biological activity was further analysed. The results are as follows: glucans make significant contributions to immunomodulation, antioxidant activity, and gut microbiota regulation; galactans are crucial for antioxidant effects, immunomodulation, and gut microbiota regulation; mannans play a key role in immunomodulation, antitumor activity, and neuroprotection; fructans are vital for gut microbiota regulation, immunomodulation, and antioxidant effects; and pectins exhibit notable immunomodulatory, antioxidant, and hypoglycaemic properties. Consequently, developing polysaccharides from medicinal plant resources based on their monosaccharide composition is expected to speed up the search for polysaccharides with high biological activity and provide a theoretical reference for polysaccharide research.

Pleurotus ostreatus is one of the most popular edible mushrooms worldwide and has been proposed as a candidate for human selenium (Se) supplementation. The objective of this study is to develop a Se-biofortification method by liquid fermentation and evaluate the effect of 2,4-dichlorophenoxyacetic acid (2,4-D), Na2SeO3 fortification doses and initial pH on P. ostreatus mycelium growth, nutritional value, and mineral accumulation. The optimal growth conditions were identified as 2,4-D 2.00 mg L- 1 , Na2SeO3 30.65 mg L- 1 , and initial pH 5.97 by BoxBehnken design. The optimization resulted in the biomass of 15.75 g L- 1 and the organic Se content of 594.85 mg kg- 1 , which was 2.50 and 8.03 times higher than the control group, respectively. In parallel, the contents of polysaccharides, proteins, essential amino acids, major functional components, and several minerals in mycelium were increased to varying degrees. Overall, this study will provide an experimental basis for the use of P. ostreatus mycelium as a good carrier for organic Se enrichment by liquid fermentation and technical support for the industrial development of new Se biofortification methods.

3′-sialyl-N-acetyllactosamine (3′-SLN) and 6′-sialyl-N-acetyllactosamine (6′-SLN) are two important human milk oligosaccharides (HMOs) which play significant functions in brain development and antiviral potential. However, their metabolism is still unknown. In this study, chemoenzymatically synthesized 3′-SLN and 6′-SLN were labeled with cyanine-7 (Cy7) via formation triazole (Tz) derivatives to investigate their metabolism and organ distribution in a mouse model. The fluorescence signals were detected in the brains of mice after 0.5 h of gavage with 3′-SLN-Tz-Cy7 and 6′-SLN-Tz-Cy7. It was found for the first time that both of them can cross the blood–brain barrier (BBB) as a whole and reach the brain in a sex-specific manner. And the results show that, whether in the in vivo imaging results or the brain fluorescence signal results, the male mice absorbed 3′-SLN-Tz-Cy7 more than 6′-SLN-Tz-Cy7; meanwhile, in the female mice, the results were exactly the opposite. Both 3′-SLN-Tz-Cy7 and 6′-SLN-Tz-Cy7 exhibited the highest fluorescence intensity in pulmonary tissues, followed by substantial hepatic deposition. This study would offer preliminary evidence for the hypothesis that the oral administration of 3′-SLN and 6′-SLN may promote brain development and provide a foundation for the further exploration of their functions in brain cognition.

The increasing challenges of pesticide resistance and environmental degradation in modern agriculture require sustainable pest management strategies. This study investigates the efficacy of a specific type of hetero-chitooligosaccharide called DACOS, as a plant defense inducer against the two-spotted spider mite, Tetranychus urticae Koch. DACOS is characterized with 85% deacetylation and an average molecular weight of 1 kDa. In planta bioassays revealed a dose-dependent suppression of mite performance on DACOS-treated leaves of kidney bean seedlings, with peak efficacy occurring at 80 ppm. This concentration reduced mite survival by 30% and fecundity by 59% after six days. In addition to the local effects, mite survival and fecundity were also reduced on untreated leaves of the same seedlings, confirming that the effect of DACOS on plant defense is systemic. Behavioral choice assays revealed that DACOS-treated leaves induced 73% mite repellency. No direct acaricidal effects were observed, which confirms the role of DACOS as a plant defense elicitor. Metabolic profiling revealed enriched pathways (e.g., monoterpenoid biosynthesis and α-linolenic acid metabolism) that produce volatile organic compounds with repellent properties. These findings suggest that DACOS is an environmentally friendly biostimulant that can enhance plant defense, bypass pesticide resistance, and promote the sustainability of crop production. ### Competing Interest Statement The authors have declared no competing interest. JSPS KAKENHI, 24K21256 National Key R&D Program of China, 2024YFD2402105

In this study, a homogeneous polysaccharide component, namely SREP-1, was purified from Stropharia rugosoannulata fermentation broth. SREP-1 was identified as a novel water-soluble neutral polysaccharide, with a molecular weight of 9.6 kDa. Monosaccharide composition analysis showed that SREP-1 was composed of glucose, galactose and mannose in a molar ratio of 78.6: 13.6: 7.8. The primary structure was elucidated through FT-IR, methylation analysis and NMR spectroscopy, revealing a backbone of →4)-α-D-Glcp-(1 → and →4,6)-α-D-Glcp-(1 → residues, and →6)-α-D-Galp-(1→, β-D-Manp-(1 → and α-D-Glcp-(→1 residues for the branched chains. Results indicated that SREP-1 possessed an amorphous globular-like structure, good thermally stability and triple-helix conformation in water. In vivo results showed that SREP-1 reversed D-galactose (D-gal)-induced body weight and organ indexes decrease, and alleviated liver damage according to improved histopathology and declined indicators in serum. Amelioration of oxidative stress and abnormal inflammation of aging liver might be due to the elevated nuclear factor-erythroid 2-related factor 2 (Nrf2) expression and decreased that of nuclear factor-κB p65 (NF-κB p65). Interestingly, the beneficial effects of SREP-1 were abolished after pretreatment with antibiotics. Our findings demonstrated that the role of SREP-1 in attenuating aging-related liver injury might involve the regulation of Nrf2-NF-κB signaling pathway and its prebiotic effect.

Stropharia rugosoannulata (S. rugosoannulata) is one of the most widely recognized edible mushrooms globally. However, knowledge of the fermentation process remains limited. In this study, a combination of statistical optimization techniques was employed to optimize the culture medium for exopolysaccharide (SREP) production during submerged fermentation. These methods included one-factor-at-a-time experiments, Plackett-Burman design, the steepest ascent method, and the Box-Behnken design. Maltose, pH and inoculum size were identified as the most significant factors affecting SREP production. The optimized conditions were determined to be: potato 150 g/L, maltose 26.8 g/L, yeast paste 1 g/L, KH2PO4 0.5 g/L, MgSO4 0.5 g/L, vitamin C 0.02 g/L, temperature 28 degrees C, rotational speed 150 r/min, inoculum size 7 %, liquid volume 200 mL, and initial pH 9. Compared to pre-optimization conditions, the yield of SREP increased from 1.06 g/L to 1.32 g/L following optimization. Additionally, cultivation kinetic models were established to study the relationships between mycelial cell growth, SREP production and substrate consumption throughout the fermentation process. These models provided practical guidance for the production of exopolysaccharides from S. rugosoannulata through liquid submerged fermentation.

Human milk oligosaccharides (HMOs) play important roles in the development of infants, which are the third most abundant component in human milk. N-Acetyllactosamine (LacNAc) is an important intermediate for the biosynthesis of other HMOs and antigens. Since currently appropriate synthetic methods for large-scale production of LacNAc are not available, it is urgently needed to develop an efficient and cost-effective synthetic pathway for LacNAc preparation. In this study, a cost-effective pathway of LacNAc synthesis involving regeneration of adenosine triphosphate (ATP) and uridine 5'-triphosphate (UTP) was established. After optimizing the reaction conditions, LacNAc was synthesized at a yield of >90% via a sequential one-pot multienzyme (OPME) method with crude enzymes at 100 mM substrates. Finally, LacNAc was produced efficiently and cost-effectively at a 5 L scale. This strategy would possibly meet the requirements of potential industrial production of LacNAc and would provide guidance for the production of other structurally complex HMOs or functional oligosaccharides in the future.

Glycans are one of the most important biomacromolecules and are essential to nearly all known life forms. However, their structural complexity makes structural analysis more challenging than nucleic acids or proteins, significantly limiting the opportunities for investigating their structure-function relationship. Here, we proposed a novel glycan sequencing strategy in which a suite of specific non-reducing-end exoglycosidases was integrated with electrochemical detection of monosaccharides in a digital microfluidic (DMF) chip. In this study, a DMF platform with integrated non-enzymatic electrochemical sensors for monosaccharides was set up and characterized. Nine mammal monosaccharides were electrochemically investigated for the first time through cyclic voltammetry (CV) and pulse amperometry (PA) on this DMF platform. Automatic enzymatic hydrolysis of tagged lactoses was performed, and released monosaccharides were electrochemically detected and quantified, and the sequence of lactose was deduced. This current study would offer a promising new strategy for accurate and efficient sequencing of pure glycans with defined sequences and connectivity in their molecular structures.

Alginate oligosaccharide (AOS) is a structurally distinct carbohydrate derived from marine algae. In this study, AOS was obtained through the enzymatic hydrolysis of alginate, and the anti-inflammatory efficacy of AOS was assessed in lipopolysaccharide (LPS)-induced inflammatory Balb/c mice. AOS effectively suppressed the overexpression of TNF-α, IL-6, and MDA while restoring the reduced SOD activity. Histopathological analysis revealed that AOS significantly reduced the level of LPS-induced tissue edema, inflammatory infiltration, and villous destruction. Additionally, AOS notably upregulated tight junction proteins Claudin-1, Occludin, and ZO-1 expression. Transcriptomic and Western blot analyses indicated that AOS primarily mediated the restriction of the TLR4/MAPK/NF-κB pathway in the jejunum. Moreover, AOS ameliorated gut microbiota dysbiosis, such as increasing in Bacteroidota, alongside decreasing in Firmicutes, Campylobacter, and Desulfovibrio, respectively. Metabolomics demonstrated that AOS improved the LPS-induced reduction of short-chain fatty acids in the gut. These results provide compelling evidence supporting the potential of AOS against acute intestinal inflammation.

Polysaccharides are natural polymers that widely exist in medicinal plants. Polysaccharides are complex molecules composed of long chains of monosaccharide units joined by glycosidic bonds from a structural point of view. Modern pharmacological research shows that the molecular weight (MW) of polysaccharides is closely related to their bioactivity. This review summarizes the preparation and detection methods of medicinal plant polysaccharides with different molecular weights. Subsequently, the paper analyzed the general experience of the correlation between bioactivity and their relative MW of polysaccharides from medicinal plants. The results showed that the polysaccharides with large relative MW (> 100 kDa) play a leading role in immune regulation. Polysaccharides with medium relative MW (10-100 kDa) play a leading role in the protection of the liver. Polysaccharides with small relative MW (< 10 kDa) play a leading role in the antioxidant effect, regulation of intestinal flora, and regulation of blood glucose. Consequently, the precise development of polysaccharides from medicinal plant resources based on relative MW is expected to be a shortcut for searching high biological activities of polysaccharides and provide a theoretical reference for polysaccharide research.

Among over 200 human milk oligosaccharides, 6'-SLN (6'-sialyl-N-acetyllactosamine) and 3'-SLN possess potential antiviral function and can potentially enhance brain development and knowledge. So far, no efficient and cost-effective synthetic pathways for 6'-SLN and 3'-SLN have been developed. In this study, efficient enzymatic pathways to synthesize 6'-SLN and 3'-SLN were established involving economic cytidine 5'-triphosphate (CTP) regeneration from commercial N-acetyllactosamine (LacNAc) with yields of >95%. To meet the requirement of industrial production, their synthesis was further optimized starting from in situ-produced LacNAc. Consequently, the cost of LacNAc purification could be saved. Furthermore, conducting synthesis at pH 7.0 and with supplementation of polyP6 in three batches led to yields of 6'-SLN (95.0%) and 3'-SLN (94.8%), respectively. Under optimized conditions, enzymatic syntheses of 6'-SLN and 3'-SLN were conducted at a 5 L scale with yields of 96.1% (259.2 g) and 92.3% (248.7 g), respectively. This study would lay the foundation for industrial production of 3'-SLN and 6'-SLN in the future and provide a guide for multienzymatic preparation of other sialylated oligosaccharides.

ABSTRACT The exopolysaccharide SREP‐1, purified from the fermentation broth of Stropharia rugosoannulata , exhibited antiaging potential. As aging significantly alters gut structure and function, protective effect of SREP‐1 was investigated using a d ‐galactose‐induced aging mouse model. SREP‐1 administration reversed D‐galactose‐induced body weight loss and colon damage, as evidenced by improved histopathology. SREP‐1 mitigated weight loss and colon damage, enhanced the activities of antioxidants (SOD, GSH‐Px, and CAT), and reduced the level of MDA. It decreased proinflammatory cytokines (TNF‐α, IL‐1β, and IL‐6) and elevated IL‐10 in colon tissue, while boosting serum immunoglobulins (IgG and IgM). Crucially, these effects were abolished by antibiotic pretreatment, highlighting the role of gut microbiota in SREP‐1 bioactivity. This role was further confirmed through fecal microbiota transplantation (FMT) and fecal supernatant transplantation (FST) experiments. Based on 16S rRNA sequencing, SREP‐1 restored gut microbial diversity, increased beneficial genera (e.g., Faecalibacterium , Akkermansia , Lactobacillus , and Bacteroides ), and decreased harmful bacteria (e.g., Escherichia‐Shigella and Collinsella ). Furthermore, short‐chain fatty acids (SCFAs) levels were elevated in the SREP‐1 group, which might regulate GPCRs/NF‐κB/Nrf2 signaling pathways and exert biological activity. This study revealed the potential of SREP‐1 to alleviate aging‐related intestinal dysfunction and underscored the crucial role of gut microbiota in mediating these effects.