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Background: The demand for animal products is increasing in developing countries due to population growth. However, livestock production contributes significantly to global warming, accounting for 25%. Probiotics can help improve livestock efficiency by enhancing gut microbes and fat metabolism. They can modify rumen populations, enhance fermentation, reduce methane emissions and improve feed digestion. In this study, the goal was to determine the most effective method of reducing methane emissions in the rumen of sheep in vitro by adding different concentrations of Saccharomyces cerevisiae and Bacillus subtilis., Results: Adding 8 × 10 6  CFU g -1 S. cerevisiae during fermentation reduced pH levels after 48 h. This also increased the concentrations of NH 3 -N, microbial protein and total gas production. At the same time, it decreased methane emissions. Furthermore, adding 20 × 10 6  CFU g -1 B. subtilis to the mixture increased total gas production (TGP) and methane production, with the highest production observed after 48 h. However, it did not affect pH levels after 48 h., Conclusion: It can be concluded that S. cerevisiae had significantly increased microbial protein and NH 3 -N concentrations after fermentation without altering pH. Additionally, the addition of S. cerevisiae enhanced TGP and reduced methane emissions. It is worth noting that TGP increased because B. subtilis was added at a concentration of 20 × 10 6  CFU g -1 , with no significant differences between concentrations. Therefore, we recommend adding S. cerevisiae and B. subtilis to the diet at doses of 8 and 20 × 10 6  CFU g -1 , as it resulted in higher TGP and reduced methane emissions. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Two protocols involving batch cultures were used to investigate the bioaugmentation of methane production by Pecoramyces ruminantium, and Methanobrevibacter thaueri. Protocol I examined the effect of altering the proportion of the microbial constituents in inoculum on alfalfa stalk fermentations and showed a 25 % improvement in dry matter loss in cultures where the inoculum contained just 30 % of co-culture and 70 % of fungal monoculture. Protocol II involved consecutive cultures and alternating inoculations. This protocol resulted in 17-22 mL/g DM methane production with co-cultures a 30 % increase in methane relative to the fungal monoculture. Both protocols indicate that the co-culture rapidly dominated and was more resilient than the monoculture. Synergistic interaction between fungus and methanogen, promoted more efficient lignocellulose degradation and higher methane yield. This study highlighted the potential of microbial co-cultures for enhancing methane production from lignocellulosic biomass, offering a promising bioaugmentation strategy for improving biogas yields and waste valorization., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Lead is a common environmental pollutant which can accumulate in the kidney and cause renal injury. However, regulatory effects and mechanisms of Sparassis latifolia polysaccharide (SLP) on lipid metabolism abnormality in kidney exposed to lead are not clarified. In this study, mice were used to construct an animal model to observe the histopathological changes in kidney, measure lead content, damage indicators, differentially expressed metabolites (DEMs) and genes (DEGs) in key signaling pathways that cause lipid metabolism abnormalities based on lipidomics and transcriptomics, which were later validated using qPCR and western blotting. Co-treatment of Pb and N-acetylcysteine (NAC) were used to verify the link between SLP and oxidative stress. Our results indicated that treatment with SLP identified 276 DEMs (including metabolism of glycerophospholipid, sphingolipid, glycerolipid and fatty acid) and 177 DEGs (including genes related to oxidative stress, inflammation, autophagy and lipid metabolism). Notably, regulatory effects of SLP on abnormal lipid metabolism in kidney were mainly associated with oxidative stress, inflammation and autophagy; SLP could regulate abnormal lipid metabolism in kidney by reducing oxidative stress and affecting its downstream-regulated autophagy and inflammatory to alleviate renal injury caused by lead exposure. This study provides a theoretical basis for SLP intervention in lead injury., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Morchella importuna polysaccharide (MIP) has been proven to have obvious hypoglycemic effects on mice with type 2 diabetes (T2DM). This study looked at the functional and rheological characteristics of MIP, and investigated the effects of MIP on the human fecal microbiota through in vitro fermentation experiments. The outcomes demonstrate the excellent oil-holding capacity, emulsifying, foaming, and rheological characteristics of MIP. After salivary gastrointestinal digestion, the Mw of MIP decreased from 398.2 kDa and 21.5 kDa to 21.9 kDa and 11.7 kDa. By 16S rRNA sequencing of bacteria fermented in vitro, it was found that MIP did not improve the richness and diversity of intestinal microorganisms, but it may exert an anti-T2DM function by significantly increasing the relative abundance of Firmicutes and promoting Ruminococcaceae_UCG_014 , Bacteroides , and Blautia proliferation. Escherichia-Shigella could also be inhibited to improve the intestinal microenvironment. In addition, the fermentation of MIP increased the total short-chain fatty acid (SCFA) concentration from 3.23 mmol/L to 39.12 mmol/L, and the propionic acid content increased significantly. In summary, MIP has excellent processing performance and is expected to exert potential anti-T2DM activity through the human intestinal microbiota, which has broad market prospects.

Background: Amauroderma rugosum (AR) is a medicinal mushroom commonly used to treat inflammation, gastric disorders, epilepsy, and cancers due to its remarkable anti-inflammatory and anti-oxidative properties. This study was designed to evaluate the pharmacological effects of AR and its underlying mechanism of action against ulcerative colitis (UC) in vitro and in vivo., Methods: A UC mouse model was established by administration of dextran sulfate sodium (DSS). AR extract was administered intragastrically to mice for 7 days. At the end of the experiment, histopathology, macrophage phenotype, oxidative stress, and inflammatory status were examined in vivo. Furthermore, RAW 264.7, THP-1, and Caco-2 cells were used to elucidate the mechanism of action of AR in vitro., Results: AR extract (0.5-2 mg/mL) significantly suppressed lipopolysaccharide (LPS) and interferon-gamma (IFN-γ)-induced M1 macrophage (pro-inflammatory) polarization in both RAW 264.7 and THP-1 cells. LPS-induced pro-inflammatory mediators (nitric oxide, TNF-α, IL-1β, MCP-1, and IL-6) were reduced by AR extract in a concentration-dependent manner. Similarly, AR extract downregulated MAPK signaling activity in LPS-stimulated RAW 264.7 cells. AR extract elicited a concentration-dependent increase in the mRNA expression of M2 (anti-inflammatory) phenotype markers (CD206, Arg-1, Fizz-1, and Ym-1) in RAW 264.7 cells. Moreover, AR extract suppressed DSS-induced ROS generation and mitochondrial dysfunction in Caco-2 cells. The in vivo experiment revealed that AR extract (200 mg/kg) increased colon length compared to the DSS-treated group. In addition, disease activity index, spleen ratio, body weight, oxidative stress, and colonic inflammation were markedly improved by AR treatment in DSS-induced UC mice. Finally, AR suppressed M1 and promoted M2 macrophage polarization in UC mice., Conclusion: The AR extract protected against DSS-induced UC by regulating macrophage polarization and suppressing oxidative stress. These valuable findings suggest that adequate intake of AR can prevent and/or treat UC., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

To improve the functions of Pleurotus ostreatus polysaccharide (POP), POP-EGCG conjugates were prepared using free radical graft polymerization reactions and were characterized using UV-vis, FT-IR, SEM, XRD, DSC, TG, particle size and potential, three-phase contact angle, and rheological tests; The antioxidant and antibacterial ability in vitro were detected. Moreover, effects of POP-EGCG on the quality of refrigerated minced pork were investigated. The results showed the optimal preparation conditions of POP-EGCG were 1 % POP, 1.3 % EGCG, 0.25 % Vc, 16 % concentration of H 2 O 2 , and reaction 17 h. The POP-EGCG showed the characteristic peak of EGCG and was a mesh honeycomb with rough and porous surface; It had higher crystallinity, increased particle size, but decreased thermal stability, solubility, and viscosity, and significantly enhanced antioxidant and antibacterial ability. The POP-EGCG effectively improved the sensory quality and inhibited lipid oxidation of chilled minced pork, and extended the shelf life of minced pork up to 9 days at 4 °C. Specifically, the TVB-N and TBARS of minced pork in the POP-EGCG group were respectively 14.93 mg/100 g and 0.9 mg MDA/kg, which were lower than the spoilage thresholds in the national standard. This study provides a theoretical basis for further development of natural antioxidants and antimicrobial agents., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appea red to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Objective: Previous research reported that dietary addition with phytosterols improved the energy utilisation of the rumen microbiome, suggesting its potential to alleviate the negative energy balance of perinatal cows. This experiment aimed to explore the effects of feeding phytosterols on the metabolic status of perinatal cows through plasma metabolomics and faecal bacteria metabolism., Methods: Ten perinatal Holstein cows (multiparous, 2 parities) with a similar calving date were selected four weeks before calving. After 7 days for adaptation, cows were allocated to two groups (n = 5), which respectively received the basal rations supplementing commercial phytosterols at 0 and 200 mg/d during a 42-day experiment. The milk yield of each cow was recorded daily after calving. On days 1 and 42, blood and faeces samples were all collected from perinatal cows before morning feeding for analysing plasma biochemicals and metabolome, and faecal bacteria metabolism., Results: Dietary addition with phytosterols at 200 mg/d had no effects on plasma cholesterol and numerically increased milk yield by 1.82 kg/d (p>0.10) but attenuated their negative energy balance in perinatal cows as observed from the significantly decreased plasma level of β-hydroxybutyric acid (p = 0.002). Dietary addition with phytosterols significantly altered 12 and 15 metabolites (p<0.05) within the plasma and faeces of perinatal cows, respectively. Of these metabolites, 5 upregulated plasma fatty acids indicated an improved energy status (i.e., C18:1T, C14:0, C17:0, C18:0, and C16:0). Milk yield negatively correlated with plasma concentrations of ketone bodies (p = 0.035) and 5-methoxytryptamine (p = 0.039). Furthermore, dietary addition with phytosterols at 200 mg/d had no effects on fermentation characteristics and bacterial diversity of cow faeces (p>0.10) but improved potentially beneficial bacteria such as Christensenellaceae family (p<0.05) that positively correlated with feed efficiency., Conclusion: Dietary addition with phytosterols at 200 mg/d could effectively improve the energy status in perinatal cows by attenuating their negative energy balance.

As a crucial component of the fungal cell membranes, ergosterol has been demonstrated to possess surface activity attributed to its hydrophobic region and polar group. However, further investigation is required to explore its emulsification behavior upon migration to the oil-water interface. Therefore, this study was conducted to analyze the interface properties of ergosterol as a stabilizer for water in oil (W/O) emulsion. Moreover, the emulsion prepared under the optimal conditions was utilized to load the water-soluble bioactive substance with the chlorogenic acid as the model molecules. Our results showed that the contact angle of ergosterol was 117.017°, and its dynamic interfacial tension was obviously lower than that of a pure water-oil system. When the ratio of water to oil was 4: 6, and the content of ergosterol was 3.5 % (ergosterol/oil phase, w/w), the W/O emulsion had smaller particle size (438 nm), higher apparent viscosity, and better stability. Meanwhile, the stability of loaded chlorogenic acid was improved under unfavorable conditions (pH 1.2, 90 °C, ultraviolet irradiation, and oxidation), which were 73.87 %, 59.53 %, 62.53 %, and 69.73 %, respectively. Additionally, the bioaccessibility of chlorogenic acid (38.75 %) and ergosterol (33.69 %), and the scavenging rates of the emulsion on DPPH radicals (81.00 %) and hydroxyl radicals (82.30 %) were also enhanced. Therefore, a novel W/O Pickering emulsion was prepared in this work using ergosterol as an emulsifier solely, which has great potential for application in oil-based food and nutraceutical formulations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

This study aimed to develop novel nanoparticles that can serve as an excellent oil-in-water (O/W) Pickering stabilizer. The polysaccharide-protein complex nanoparticles (PPCNs-20 and PPCNs-40) were prepared at different ultrasonication amplitudes (20 % and 40 %, respectively) from the polysaccharide-protein complexes (PPCs) which were extracted from the residue of Clitocybe squamulose. Compared with PPCs and PPCNs-20, the PPCNs-40 exhibited dispersed blade and rod shape, smaller average size, and larger zeta potential, which indicated significant potential in O/W Pickering emulsion stabilizers. Subsequently, PPCNs-40 stabilized Pickering emulsions were characterized at different concentrations, pHs, and oil phase contents. The average size, micromorphology, rheological properties, and storage stability of the emulsions were improved as the concentration of PPCNs-40, the ratio of the soybean oil phase and pH value increased. Pickering emulsions showed the best stability when the concentration of PPCNs-40 was 3 wt%, and the soybean oil fraction was 30 % under both neutral and alkaline conditions. The emulsions demonstrated shear thinning and gelation behavior. These findings have implications for the use of eco-friendly nanoparticles as stabilizers for Pickering emulsions and provide strategies for increasing the added value of C. squamulosa., Competing Interests: Declaration of competing interest We confirm that there are not any conflicts of interest with respect to this publication and there has been no financial support for this article that could influence the outcome., (Copyright © 2024. Published by Elsevier B.V.)

Amauroderma rugosum (AR), also known as "Blood Lingzhi" in Chinese, is a basidiomycete belonging to the Ganodermataceae family. Four polysaccharide fractions were systematically isolated and purified from AR. Subsequently, their compositions were examined and analyzed via high-performance gel permeation chromatography (HPGPC), analysis of the monosaccharide composition, Fourier-transform infrared spectroscopy (FT-IR), and 1 H nuclear magnetic resonance (NMR). The zebrafish model was then used to screen for proangiogenic activities of polysaccharides by inducing vascular insufficiency with VEGF receptor tyrosine kinase inhibitor II (VRI). The third fraction of AR polysaccharides (PAR-3) demonstrated the most pronounced proangiogenic effects, effectively ameliorating VRI-induced intersegmental vessel deficiency in zebrafish. Concurrently, the mRNA expression levels of vascular endothelial growth factor (VEGF)-A and VEGF receptors were upregulated by PAR-3. Moreover, the proliferation, migration, invasion, and tube formation of human umbilical vein endothelial cells (HUVECs) were also stimulated by PAR-3, consistently demonstrating that PAR-3 possesses favorable proangiogenic properties. The activation of the Akt, ERK1/2, p38 MAPK, and FAK was most likely the underlying mechanism. In conclusion, this study establishes that PAR-3 isolated from Amauroderma rugosum exhibits potential as a bioresource for promoting angiogenesis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Chemo-photodynamic combination therapy has demonstrated significant potential in the treatment of cancer. Triptolide (TPL), a naturally derived anticancer agent, when combined with the photosensitizer Chlorin e6 (Ce6), has shown to provide enhanced anti-tumor benefits. However, the development of stimuli-responsive nanovehicles for the co-delivery of TPL and Ce6 could further enhance the efficacy of this combination therapy., Methods: In this study, we synthesized a pH/ROS dual-responsive mPEG- TK -PBAE copolymer, which contains a pH-sensitive PBAE moiety and a ROS-sensitive thioketal (TK) linkage. Through a self-assembly process, TPL and Ce6 were successfully co-loaded into mPEG- TK -PBAE nanoparticles, hereafter referred to as TPL/Ce6 NPs. We evaluated the pH- and ROS-sensitive drug release and particle size changes. Furthermore, we investigated both the in vitro suppression of cellular proliferation and induction of apoptosis in HepG2 cells, as well as the in vivo anti-tumor efficacy of TPL/Ce6 NPs in H22 xenograft nude mice., Results: The mPEG- TK -PBAE copolymer was synthesized through a one-pot Michael-addition reaction and successfully co-encapsulated both TPL and Ce6 by self-assembly. Upon exposure to acid pH values and high ROS levels, the payloads in TPL/Ce6 NPs were rapidly released. Notably, the abundant ROS generated by the released Ce6 under laser irradiation further accelerated the degradation of the nanosystem, thereby amplifying the tumor microenvironment-responsive drug release and enhancing anticancer efficacy. Consequently, TPL/Ce6 NPs significantly increased PDT-induced oxidative stress and augmented TPL-induced apoptosis in HepG2 cells, leading to synergistic anticancer effects in vitro. Moreover, administering TPL/Ce6 NPs (containing 0.3 mg/kg of TPL and 4 mg/kg of Ce6) seven times, accompanied by 650 nm laser irradiation, efficiently inhibited tumor growth in H22 tumor-bearing mice, while exhibiting lower systemic toxicity., Conclusion: Overall, we have developed a tumor microenvironment-responsive nanosystem for the co-delivery of TPL and Ce6, demonstrating amplified synergistic effects of chemo-photodynamic therapy (chemo-PDT) for hepatocellular carcinoma (HCC) treatment., Competing Interests: The authors report no conflicts of interest in this work., (© 2024 Huang et al.)

This experiment aimed to examine the impact of Salicornia europaea L. extract on sheep growth performance, rumen fermentation variables, nutrient apparent digestibility, and gastrointestinal microbial diversity. Forty-eight male Altay sheep, weighing 32.5 ± 2.8 kg and approximately 3.5 months old, were chosen. Four dietary treatments, each consisting of four replicates and three sheep per replicate, were distributed randomly to the sheep. The pelleted total mixed ration containing Salicornia europaea L. extract at 0.0, 0.2, 0.4, and 0.6% DM was freely available to the sheep in the four treatment groups. The 56-day experiment consisted of 45 days of measurements followed by 11 days of adaptation. The growth performance was not affected by nutrition Salicornia europaea L. extract ( p  ≤ 0.05), but the feed-to-gain ratio was reduced when the extract was given at 0.4% DM ( p  ≤ 0.05). Compared to the 0 and 0.2% treatments, the apparent digestibility of DM, OM, NDF, and ADF was substantially greater in the 0.4, and 0.6% treatments. Furthermore, compared to sheep in the 0 and 0.2% groups, sheep in the 0.6% group had a noticeably higher apparent digestibility of CP. As the amount of Salicornia europaea L. extract added to the rumen fluid rose, the molar ratio of acetic acid increased. In contrast, the molar ratio of propionic acid gradually decreased, and the total volatile fatty acid content gradually reduced. Thus, adding a suitable quantity of Salicornia europaea L. extract to the sheep ration is natural and secure, which may improve the environmental sustainability of small ruminant production systems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Kamal, Lele, Shuzhen, Jiandi, Rongyan, Yanfeng, Wenqi, Xiangyu and Cheng.)

In this study, we used fresh Oudemansiella raphanipes as raw materials and pre-treated through hot air drying (HD), infrared radiation drying (ID), and vacuum freeze drying (VD) to investigate the effects of different drying methods on the rehydration rate, appearance quality, microstructure, and volatile flavor components of the dried products, as well as to determine the physicochemical properties and bioactivities of the polysaccharides in the dried O. raphanipes. The results showed that the VD O. raphanipes had the highest rehydration rate and the least shrinkage in appearance, and it better maintained the original color of the gills, but their aroma was not as strong as that of the HD samples. The scanning electron microscopy results indicate that VD maintains a good porous structure in the tissue, while HD and ID exhibit varying degrees of shrinkage and collapse. Seventy-five common volatile substances were detected in the three dried samples, mainly alkanes, alcohols, and esters. The polysaccharides (PS-H, PS-I, and PS-V) extracted from the dried samples of these three species of O. raphanipes had similar infrared spectral features, indicating that their structures are basically consistent. The highest yield was obtained for PS-V, and the polysaccharide content and glucuronic acid content of PS-I were higher than those of the remaining two polysaccharides. In addition, PS-V also showed better antioxidant activity and inhibitory activity against α-glucosidase as well as α-amylase. In conclusion, among the above three drying methods, the quality of O. raphanipes obtained by vacuum freeze drying is the best, and this experiment provides a theoretical basis for the selection of drying methods for O. raphanipes .

To overcome the difficulty of separation and low rate of extraction caused by highly viscous polysaccharides from Naematelia aurantialba (NA), four N. aurantialba polysaccharides (NAPs) were sequentially extracted using water (enzyme-/ultrasound-assisted extraction), alkali (0.1 mol/L NaOH), and acid (0.1 mol/L HCl), and named E-NAP, U-NAP, Al-NAP, and Ac-NAP. The properties of four NAPs were different. The yields of NAPs were 26.05 % (Ac-NAP) > 20.33 % (Al-NAP) > 17.99 % (U-NAP) > 12.77 % (E-NAP), respectively. The monosaccharide composition of NAPs was composed primarily of mannose, xylose, glucose, glucuronic acid, and galactose. Sequential extraction improved the purity and solubility of NAPs, but decreased the particle size, thermal stability, water retention, and crystallinity. Two polysaccharides, U-NAP and Al-NAP, had a triple helix structure. All the NAPs were pseudoplastic fluids with concentration/frequency-dependent entangled structure. Al-NAP with the highest viscosity exhibited an elastic gel, while Ac-NAP with the lowest viscosity was a viscous gel. The behavior of NAPs differed from that predicted using the Cox-Merz rule, and in particular, E-NAP and U-NAP more significantly deviated from the rule. In this study, four NAPs with different properties were extracted sequentially, which provided a theoretical basis for the down-stream processing with high added-value and utilization of NA and NAP., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

The Microbiome Protocols eBook (MPB) serves as a crucial bridge, filling gaps in microbiome protocols for both wet experiments and data analysis. The first edition, launched in 2020, featured 152 meticulously curated protocols, garnering widespread acclaim. We now extend a sincere invitation to researchers to participate in the upcoming 2nd version of MPB, contributing their valuable protocols to advance microbiome research., Competing Interests: The authors declare no conflict of interest., (© 2024 The Authors. iMeta published by John Wiley & Sons Australia, Ltd on behalf of iMeta Science.)

To study the gel-forming properties of polysaccharide from the fruiting body of Clitocybe squamulosa (CSFP) and its degradation product (UH-CSFP), the changes in steady-state and dynamic rheological properties of CSFP and UH-CSFP under different conditions (polysaccharide mass fraction, temperature, pH, and salt ion concentration) were studied. Polysaccharides with good gel-forming properties were selected and mixed with common edible thickeners (gelatin, guar gum, and locust bean gum), after which the properties of the composite gel were assessed. The steady-state rheological results showed that CSFP and UH-CSFP were pseudoplastic fluids, their apparent viscosity decreased with increasing temperature, the viscosity was greatest when the pH was 7. The addition of Na + and Ca 2+ could increase the viscosity, and the viscosity of UH-CSFP was lower than that of CSFP at the same mass fraction. The results of dynamic rheology indicated that G´ and G´´ of CSFP and UH-CSFP increased with increasing mass fraction, pH, and ion concentration (0.01 M to 1 M), and G´´ was always smaller than G´ indicating weak gel behavior. The thixotropy-related experimental results showed that the thixotropy ring area of CSFP and UH-CSFP increased with increasing mass fraction, the ring area of CSFP was larger than that of UH-CSFP, and the gel strength of CSFP was greater than that of UH-CSFP. The results of CSFP and three types of edible gels showed that the composite gels were pseudoplastic fluids, and their apparent viscosity was ranked (in descending order) as follows: guar bean gum, locust bean gum, and gelatin. The addition of CSFP improved the gel-forming properties of guar gum but did not significantly improve the gel properties of locust bean gum and gelatin. This study provides a theoretical basis for the selection of processing methods and the application of polysaccharides., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Background: Providing high-quality roughage is crucial for improvement of ruminant production because it is an essential component of their feed. Our previous study showed that feeding bio-fermented rice straw (BF) improved the feed intake and weight gain of sheep. However, it remains unclear why feeding BF to sheep increased their feed intake and weight gain. Therefore, the purposes of this research were to investigate how the rumen microbiota and serum metabolome are dynamically changing after feeding BF, as well as how their changes influence the feed intake, digestibility, nutrient transport, meat quality and growth performances of sheep. Twelve growing Hu sheep were allocated into 3 groups: alfalfa hay fed group (AH: positive control), rice straw fed group (RS: negative control) and BF fed group (BF: treatment). Samples of rumen content, blood, rumen epithelium, muscle, feed offered and refusals were collected for the subsequent analysis., Results: Feeding BF changed the microbial community and rumen fermentation, particularly increasing (P < 0.05) relative abundance of Prevotella and propionate production, and decreasing (P < 0.05) enteric methane yield. The histomorphology (height, width, area and thickness) of rumen papillae and gene expression for carbohydrate transport (MCT1), tight junction (claudin-1, claudin-4), and cell proliferation (CDK4, Cyclin A2, Cyclin E1) were improved (P < 0.05) in sheep fed BF. Additionally, serum metabolome was also dynamically changed, which led to up-regulating (P < 0.05) the primary bile acid biosynthesis and biosynthesis of unsaturated fatty acid in sheep fed BF. As a result, the higher (P < 0.05) feed intake, digestibility, growth rate, feed efficiency, meat quality and mono-unsaturated fatty acid concentration in muscle, and the lower (P < 0.05) feed cost per kg of live weight were achieved by feeding BF., Conclusions: Feeding BF improved the growth performances and meat quality of sheep and reduced their feed cost. Therefore, bio-fermentation of rice straw could be an innovative way for improving ruminant production with minimizing production costs., (© 2024. The Author(s).)

Biomass from agriculture, forestry, and urban wastes is a potential renewable organic resource for energy generation. Many investigations have demonstrated that anaerobic fungi and methanogens could be co-cultured to degrade lignocellulose for methane generation. Thus, this study aimed to evaluate the effect of natural anaerobic fungi-methanogens co-culture on the methane production and lignocellulosic degradation of wastes from rice, corn and sugarcane. Hu sheep rumen digesta was used to develop a natural anaerobic fungi-methanogen co-culture. The substrates were rice straw (RS), rich husk (RH), corn stover (CS), corn cobs (CC), and sugarcane baggage (SB). Production of total gas and methane, metabolization rate of reducing sugar, glucose, and xylose, digestibility of hemicellulose and cellulose, activity of carboxymethylcellulase and xylanase, and concentrations of total acid and acetate were highest (P < 0.05) in CC, moderate (P < 0.05) in RS and CS, and lowest (P < 0.05) in SB and RH. The pH, lactate and ethanol were lowest (P < 0.05) in CC, moderate (P < 0.05) in RS and CS, and lowest (P < 0.05) SB and RH. Formate was lowest (P < 0.05) in CC, RS and CS, moderate (P < 0.05) in SB, and lowest (P < 0.05) in RH. Therefore, this study indicated that the potential of methane production and lignocellulosic degradation by natural anaerobic fungi-methanogens co-culture were highest in CC, moderate in RS and CS, and lowest in SB and RH., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

This study comparatively evaluated the effects of the commonly used six extraction methods (acidic, alkaline, enzymatic, ultrasonic, high-pressure, and microwave) on the physico-chemical properties, processing characteristics, and biological activities of polysaccharides from Clitocybe squamulosa (CSFPs). The results show that polysaccharides extracted using an enzyme-assisted extraction method has a relatively high extraction yield (4.46 ± 1.62 %) and carbohydrate content (70.79 ± 6.25 %) compared with others. Furthermore, CSFPs were all composed of glucose, galactose, mannose, xylose, and glucosamine hydrochloride. Only ultrasonic-assisted extraction of polysaccharides (CSFP-U) has a triple helix chain conformation. Scanning electron microscopy (SEM) revealed significant differences in the microstructure of polysaccharides prepared using different methods. Besides that, the polysaccharides prepared by alkali extraction (CSFP-B) and high-pressure assisted extraction (CSFP-H) have good water (2.86 ± 0.29 g/g and 3.15 ± 0.29 g/g) and oil (8.13 ± 0.32 g/g and 7.97 ± 0.04 g/g) holding properties. The rheological behavior demonstrated that CSFPs solutions were typical non-Newtonian fluid. Apart from this, the antioxidant capacity (clearing DPPH (IC 50  = 0.29) and ABTS free radicals (IC 50  = 0.19), total reduction ability (IC 50  = 3.02)) of polysaccharides prepared by the microwave-assisted extraction (CSFP-M) method was significantly higher than that of other extraction methods. By contrast, the polysaccharide prepared by acid extraction (CSFP-A) has the optimum binding capacity (bile acid salt (71.30 ± 6.78 %) and cholesterol (57.07 ± 3.26 mg/g)). The antibacterial activity of CSFPs was positively correlated with their concentration. Thus, the research results can provide a theoretical basis for the development and utilization of polysaccharides from C. squamulosa., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

To study the effects of Naematelia aurantialba (NA) on the rheological and gelatinization properties of starch, the processing methods of NA were diversified. In this study, the gelatinization and rheological properties of corn starch (CS) and edible cassava starch (ECS) were investigated by adding NA with different mass fractions. Starch soft candy was prepared using NA, CS, and ECS as the main raw materials. Rheological studies showed that both CS-NA and ECS-NA exhibited elastic modulus (G') > viscosity modulus (G″), implying elastic behavior. G' was such that CS+1%NA > CS+5%NA > CS+3%NA > CS > CS+2%NA > CS+4%NA > ECS+4%NA > ECS+3%NA > ECS+5%NA > ECS+2%NA > ECS+1%NA > ECS. The gelatinization implied showed that after adding NA, the pasting temperature of CS-NA and ECS-NA increased by 1.33 °C and decreased by 2.46 °C, while their breakdown values decreased by 442.35 cP and 866.98 cP, respectively. Through a single-factor test and orthogonal test, the best formula of starch soft candy was as follows: 0.4 f of NA, 10 g of white granulated sugar, a mass ratio of ECS to CS of 20:1 (g:g), 0.12 g of citric acid, 1 g of red date power, and 16 mL of water. The soft candy was stable when stored for two days. This study offers a new direction for the research and development of NA starch foods.

The aim of this study was to investigate the effects of ensiled agricultural byproducts from Qinghai-Tibet plateau on growth performance, rumen microbiota, ruminal epithelium morphology, and nutrient transport-related gene expression in Tibetan sheep. Fourteen male Tibetan sheep were randomly assigned to one of two diets: an untreated diet (without silage inoculum, CON, n = 7) or an ensiled diet (with silage inoculum, ESD, n = 7). The total experimental period lasted for 84 d, including early 14 d as adaption period and remaining 70 d for data collection. The ESD increased average daily gain (P = 0.046), dry matter intake (P < 0.001), ammonia nitrogen (P = 0.045), microbial crude protein (P = 0.034), and total volatile fatty acids concentration (P < 0.001), and decreased ruminal pH value (P = 0.014). The proportion of propionate (P = 0.006) and the copy numbers of bacteria (P = 0.01) and protozoa (P = 0.002) were higher, while the proportion of acetate (P = 0.028) was lower in the sheep fed ESD compared to CON. Pyrosequencing of the 16S ribosomal RNA gene revealed that ESD increased the relative abundance of Firmicutes, Ruminococcus, Lachnospiraceae&#95;AC2044&#95;group, Lachnospiraceae&#95;XPB1014&#95;group, and Christensenellaceae&#95;R-7&#95;group in the rumen (P < 0.05), while decreased the relative abundance of Bacteroidota, Prevotellaceae&#95;UCG-003, and Veillonellaceae&#95;UCG-001 (P < 0.05). Analyses with PICRUSt2 and STAMP indicated that the propionate metabolism pathway was enriched in the sheep fed ESD (P = 0.026). The ESD increased the rumen papillae height (P = 0.012), density (P = 0.036), and surface area (P = 0.001), and improved the thickness of the total epithelia (P = 0.018), stratum corneum (P = 0.040), stratum granulosum (P = 0.042), and stratum spinosum and basale (P = 0.004). The relative mRNA expression of cyclin-dependent Kinase 2, CyclinA2, CyclinD2, zonula occludens-1, Occludin, monocarboxylate transporter isoform 1 (MCT1), MCT4, sodium/potassium pump, and sodium/hydrogen antiporter 3 were higher in the rumen epithelial of sheep fed ESD than CON (P < 0.05). Conversely, the relative mRNA expressions of Caspase 3 and B-cell lymphoma-2 were lower in the sheep fed ESD than CON (P < 0.05). In conclusion, compared with an untreated diet, feeding an ensiled diet altered the rumen microbial community, enhanced nutrient transport through rumen epithelium, and improved the growth performance of Tibetan sheep., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Xanthones are among the most fundamental phytochemicals in nature. The anti-cancer activities of xanthones and their derivatives have been extensively studied. Recently, we found that garcinone E (GE), an effective anti-cancer phytochemical isolated from mangosteen (Garcinia mangostanal.), showed promising anti-cancer effects in vitro and in vivo. However, little is known about its effects on epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor 2 (VEGFR2) activity., Purpose: This study aimed to identify potent dual EGFR and VEGFR2 inhibitors from mangosteen-derived xanthones using structure-activity relationship analyses., Study Design: The interaction of xanthones with EGFR and VEGFR2 was analyzed using molecular docking experiments. The kinase activities of EGFR and VEGFR2 were determined using bioluminescence assays. The rat aortic ring and Matrigel plug angiogenesis assays were used to evaluate blood vessel formation ex vivo and in vivo. A breast tumor-bearing nude mouse model was established to examine the anti-tumor effects of different xanthones., Results: Molecular docking analysis showed that GE bound tightly to EGFR and VEGFR2, with binding energies of -9.73 and -9.56 kcal/mol, respectively. Kinase activity assessment showed that GE strongly inhibited both EGFR and VEGFR2 kinase activity, with IC 50 values of 315.4 and 158.2 nM, respectively. Moreover, GE significantly abolished the EGF- and VEGF-induced phosphorylation of EGFR and VEGFR2, respectively. GE also showed strong inhibitory effects on cancer cell growth, endothelial cell migration, invasion, and tube formation. Ex vivo and in vivo angiogenesis assays showed that GE dose-dependently suppressed blood vessel formation in the rat aorta, Matrigel plugs, and transgenic zebrafish embryos, with the lowest effective concentration of 0.25 μM. Furthermore, GE (2 mg/kg) strongly inhibited tumor growth and reduced tumor weight in MDA-MB-231 breast tumor-xenografted mice. GE significantly reduced microvessel density and downregulated the expression of VEGFR2, EGFR, and Ki67 in tumor tissues., Conclusion: The present study demonstrated that GE was the most potent dual inhibitor of EGFR and VEGFR2 among all xanthones tested. These findings may provide valuable information for the future development of novel and effective dual inhibitors of EGFR and VEGFR2., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier GmbH.)

Exposure to lead can have harmful effects on the intestines and gut microbiota, leading to toxicity. This study aimed to explore the protective role of Sparassis latifolia polysaccharide (SLP) in safeguarding the intestinal barrier of Kunming mice exposed to lead. The findings indicated that SLP effectively alleviates intestinal lesions, increases the density of cupped cells in the intestine, and reduces inflammation in both serum and the small intestine. Furthermore, SLP maintains the expression of key genes such as ZO-1, Occludin, Claudin-1, Lyz, Ang4, and ZO-2, as well as proteins like claudin-1 and Occludin-1. Furthermore, SLP positively impacts the diversity and richness of microorganisms in the mouse gut microbiota at both the genus and gate levels. It also increases the levels of short-chain fatty acids (SCFAs), including acetic acid, butyric acid, and propionic acid, to varying degrees. In summary, SLP plays a role in alleviating the impaired small intestinal barrier in lead-exposed mice by modulating the intestinal flora, which is consistent with reduced lead absorption. This modulation enhances the integrity of the intestinal barrier, suppresses inflammation, and facilitates the excretion of lead., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Cancer metastasis remains the most common cause of death in breast cancer patients. Tumor-associated macrophages (TAMs) are a novel therapeutic target for the treatment of metastatic breast cancer. Despite the good anti-cancer activity of garcinone E (GE), there are no reports on its therapeutic effects on breast cancer metastasis. The objective of this study was to examine the anti-cancer effects of GE on metastatic breast cancer. RAW 264.7 and THP-1 cells were polarized to M2 macrophages by IL-4/IL-13 in vitro. A 4T1 mouse breast cancer model and the tail vein breast cancer metastasis model were used to explore the effect of GE on breast cancer growth and metastasis in vivo. In vitro studies showed that GE dose-dependently suppressed IL-4 + IL-13-induced expression of CD206 in both RAW 264.7 cells and differentiated THP-1 macrophages. However, GE did not affect the LPS + IFN-γ-induced polarization to the M1-like macrophages in vitro. GE inhibited the expression of the M2 macrophage specific genes in RAW 264.7 cells, and simultaneously impaired M2 macrophage-induced breast cancer cell proliferation and migration, and angiogenesis. In animal studies, GE significantly suppressed tumor growth, angiogenesis, and lung metastasis in 4T1 tumor-bearing mice, without causing toxicity. In both tumor and lung tissues, the proportion of M2-like TAMs was significantly decreased while the proportion of M1-like TAMs was markedly increased by GE treatment. Mechanistically, GE inhibited phosphorylation of STAT6 in vitro and in vivo. Our results demonstrate for the first time that GE suppresses breast cancer growth and pulmonary metastasis by modulating M2-like macrophage polarization through the STAT6 signaling pathway., (© 2023 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.)

This study compared the growth performance, serum biochemical indicators, rumen fermentation parameters, rumen bacterial structure, and fecal bacterial structure of cattle and yaks fed for two months and given a feed containing concentrate of a roughage ratio of 7:3 on a dry matter basis. Compared with cattle, yak showed better growth performance. The serum biochemical results showed that the albumin/globulin ratio in yak serum was significantly higher than that in cattle. Aspartate aminotransferase, indirect bilirubin, creatine kinase, lactate dehydrogenase, and total cholesterol were significantly lower in yaks than in cattle. The rumen pH, acetate to propionate ratio, and acetate were lower in yaks than in cattle, whereas the lactate in yaks was higher than in cattle. There were significant differences in the structure of ruminal as well as fecal bacteria between cattle and yaks. The prediction of rumen bacterial function showed that there was a metabolic difference between cattle and yaks. In general, the metabolic pathway of cattle was mainly riched in a de novo synthesis of nucleotides, whereas that of yaks was mainly riched in the metabolic utilization of nutrients. This study provides a basis for understanding a rumen ecology under the condition of a high concentrate diet.

Changes in the functional properties and structures of Clitocybe squamulosa protein isolate (CSPI) in the process of freeze-thaw (F-T) cycles were explored. Remarkable alterations and the reduced content of protein ordered structure were revealed through structural analysis of CSPI after F-T treatments. The surface hydrophobicity and free sulfhydryl content of CSPI first increased and then decreased. However, after the F-T treatments, the carbonyl content of CSPI continued to increase. Similarly, the water holding capacity (WHC), oil holding capacity (OHC), and solubility of CSPI all declined as the number of F-T cycles increased. The foaming properties and emulsifying properties of CSPI were significantly improved and reached maximum values after three F-T cycles. CSPI undergoing two F-T cycles showed the highest digestibility, maximum polypeptide content, and highest DPPH and ·OH-radical-scavenging activities. The ·OH-radical-scavenging activities and reducing power of the gastrointestinally digested CSPI had the highest value after one F-T cycle. Therefore, it has been demonstrated that F-T treatments could be a residue-free and cost-effective tool for improving mushroom protein functional properties.

The microbial decomposition and utilization of lignocellulosic biomass present in the plant tissues are driven by a series of carbohydrate active enzymes (CAZymes) acting in concert. As the non-catalytic domains widely found in the modular CAZymes, carbohydrate-binding modules (CBMs) are intimately associated with catalytic domains (CDs) that effect the diverse hydrolytic reactions. The CBMs function as auxiliary components for the recognition, adhesion, and depolymerization of the complex substrate mediated by the associated CDs. Therefore, CBMs are deemed as significant biotools available for enzyme engineering, especially to facilitate the enzymatic hydrolysis of dense and insoluble plant tissues to acquire more fermentable sugars. This review aims at presenting the taxonomies and biological properties of the CBMs currently curated in the CAZy database. The molecular mechanisms that CBMs use in assisting the enzymatic hydrolysis of plant polysaccharides and the regulatory factors of CBM-substrate interactions are outlined in detail. In addition, guidelines for the rational designs of CBM-fused CAZymes are proposed. Furthermore, the potential to harness CBMs for industrial applications, especially in enzymatic pretreatment of the recalcitrant lignocellulose, is evaluated. It is envisaged that the ideas outlined herein will aid in the engineering and production of novel CBM-fused enzymes to facilitate efficient degradation of lignocellulosic biomass to easily fermentable sugars for production of value-added products, including biofuels., Competing Interests: Declaration of Competing Interest All the authors declare that they have no competing interests in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Breast cancer metastasis is leading cause of cancer death among women worldwide. Tumor-associated macrophages (TAMs) have been considered as potential targets for treating breast cancer metastasis because they promote tumor growth and development. Glycyrrhetinic acid (GA) is one of the most important phytochemicals of licorice which has shown promising anti-cancer efficacies in pre-clinical trials. However, the regulatory effect of GA on the polarization of TAMs remains elusive., Purpose: To investigate the role of GA in regulating the polarization of M2 macrophages and inhibiting breast cancer metastasis, and to further explore its underlying mechanisms of action., Study Design: IL-4 / IL-13-treated RAW 264.7 and THP-1 cells were used as the M2-polarized macrophages in vitro. A 4T1 mouse breast cancer model and the tail vein breast cancer metastasis model were applied to study the effect of GA on breast cancer growth and metastasis in vivo., Results: In vitro studies showed that GA significantly inhibited IL-4 / IL 13-induced M2-like polarization in RAW 264.7 and THP-1 macrophages without affecting M1-like polarization. GA strongly decreased the expression of M2 macrophage markers CD206 and Arg-1, and reduced the levels of the pro-angiogenic molecules VEGF, MMP9, MMP2 and IL-10 in M2 macrophages. GA also increased the phosphorylation of JNK1/2 in M2 macrophages. Moreover, GA significantly suppressed M2 macrophage-induced cell proliferation and migration in 4T1 cancer cells and HUVECs. Interestingly, the inhibitory effects of GA on M2 macrophages were abolished by a JNK inhibitor. Animal studies showed that GA significantly suppressed tumor growth, angiogenesis, and lung metastasis in BALB/c mice bearing breast tumor. In tumor tissues, GA reduced the number of M2 macrophages but elevated the proportion of M1 macrophages, accompanied by activation of JNK signaling. Similar results were found in the tail vein breast cancer metastasis model., Conclusion: This study demonstrated for the first time that GA could effectively suppress breast cancer growth and metastasis by inhibiting macrophage M2 polarization via activating JNK1/2 signaling. These findings indicate that GA could be served as the lead compound for the future development of anti-breast cancer drug., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Sparassis latifolia polysaccharides (SLPs) can regulate inflammatory cytokines. However, little is known about the regulation mechanism of SLPs on colon cancer. In this study, we investigated the mechanism of SLPs on metabolism in mice with colon cancer. The results showed that SLPs could improve the colon morphology and physiological indices, and inhibit the infiltration of immune cells in colon. Moreover, it could improve metabolism disorder of colon cancer via reducing the levels of TNF-α, IL-6, NF-κB, COX-2 and IL-1β mRNA or protein, increasing IκB mRNA or protein expression. In addition, it could comprehensively regulate the colon cancer related metabolism by changing the abundance of key intestinal flora and 35 metabolites including phosphatidylcholine, tryptophan and tetrahydrobiopterin. Some biomarkers associated with colon cancer metabolism were related significantly with the abundance of specific intestinal flora. These findings indicate that SLPs can attenuate metabolism disorder of colon cancer by modulating gut microbiota and metabolites., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The aim of this study was to establish a human digestion model in vitro to explore the degradation characteristics of a novel high-purity polysaccharide from Clitocybe squamulosa (CSFP2). The results showed that the content of reducing sugars ( C R ) of CSFP2 increased from 0.13 to 0.23 mg/mL, the molecular weight ( Mw ) of CSFP2 decreased significantly during the saliva-gastrointestinal digestion. The constituent monosaccharides of CSFP2, including galactose, glucose, and mannose, were stable during in vitro digestion, but their molar ratios were changed from 0.023: 0.737: 0.234 to 0.496: 0.478: 0.027. The surface of CSFP2 changes from a rough flaky structure to a scattered flocculent or rod-shaped structure after the gastrointestinal digestion. However, the apparent viscosity of CSFP2 was overall stable during in vitro digestion. Moreover, CSFP2 still maintains its strong antioxidant capacity after saliva-gastrointestinal digestion. The results showed that CSFP2 can be partially decomposed during digestion. Meanwhile, some physico-chemical properties of the fermentation broth containing CSFP2 changed significantly after gut microbiota fermentation. For example, the pH value (from 8.46 to 4.72) decreased significantly ( p  < 0.05) after 48 h of fermentation. the OD 600 value increased first and then decreased (from 2.00 to 2.68 to 1.32) during 48-h fermentation. In addition, CSFP2 could also increase the amounts of short-chain fatty acids (SCFAs) (from 5.5 to 37.15 mmol/L) during fermentation (in particular, acetic acid, propionic acid, and butyric acid). Furthermore, the relative abundances of Bacteriodes , Bifidobacterium , Catenibacterium , Lachnospiraceae&#95;NK4A136&#95;group , Megasphaera , Prevotella , Megamonas , and Lactobacillus at genus level were markedly increased with the intervention of CSFP2. These results provided a theoretical basis for the further development of functional foods related to CSFP2., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Background: The nutrient availability of roughages could affect the dietary utilization efficiency of ruminants even in isocaloric and isonitrogenous diets. Here, we analyzed the bacterial composition and their metabolic pathways in the gastrointestinal tracts (GITs) of Hu sheep fed with wheat straw (WS) instead of alfalfa (AL) in isocaloric and isonitrogenous diets, trying to explore the reasons from the perspective of GITs bacterial network structure changes., Results: We employed 16S rRNA gene sequencing in combination with the Kruskal-Wallis test, Spearman correlation analysis, and other statistical methods to describe the microbiota composition in the GITs of Hu sheep. The results showed after the roughage was replaced from AL to WS, the most positive response occurred in the rumen microbiota, resulting in a more obvious microbiological and functional redundancy phenomenon. Whereas extended biogeographic studies of the GITs bacterial community found opposite results for the hindgut microbiota and metabolism networks compared to the forestomach. The abundance of fiber-degrading bacteria such as Prevotella, Oscillospiraceae NK4A214 group, and Treponema was significantly increased in GITs, but low-efficiency crude fiber degradation inhibited energy use efficiency, the pentose phosphate pathway, gluconeogenesis, and volatile acid synthesis. In addition, dietary shifting from AL to WS decreased the abundance of beneficial bacteria such as the Lachnospiraceae NK3A20 group and Alistipes, thereby enhancing the underlying inflammatory response., Conclusions: These findings suggest that feeding untreated WS affected the structure and function of the bacterial network in the GITs due to limited total digestible nutrients, and in particular increases the complexity of the rumen bacterial network, and limit the abundance of bacteria involved in the crude fiber degradation in the hindgut., (© 2023. The Author(s).)

Background: Lignocellulose biomass is the most abundant and renewable material in nature. The objectives of this study were to characterize two endoglucanases TrepCel3 and TrepCel4, and determine the effect of the combination of them (1.2 mg TrepCel3, 0.8 mg TrepCel4) on in vitro rumen fermentation characteristics. In this study, three nature lignocellulosic substrates (rice straw, RS; wheat straw, WS; leymus chinensis, LC) were evaluated for their in vitro digestibility, gas, NH 3 -N and volatile fatty acid (VFA) production, and microbial protein (MCP) synthesis by adding enzymatic combination., Methods: Two endoglucanases' genes were successfully expressed in Escherichia coli (E. coli) BL21 (DE3), and enzymatic characteristics were further characterized. The combination of TrepCel3 and TrepCel4 was incubated with lignocellulosic substrates to evaluate its hydrolysis ability., Results: The maximum enzymatic activity of TrepCel3 was determined at pH 5.0 and 40 °C, while TrepCel4 was at pH 6.0 and 50 °C. They were stable over the temperature range of 30 to 60 °C, and active within the pH range of 4.0 to 9.0. The TrepCel3 and TrepCel4 had the highest activity in lichenan 436.9 ± 8.30 and 377.6 ± 6.80 U/mg, respectively. The combination of TrepCel3 and TrepCel4 exhibited the highest efficiency at the ratio of 60:40. Compared to maximum hydrolysis of TrepCel3 or TrepCel4 separately, this combination was shown to have a superior ability to maximize the saccharification yield from lignocellulosic substrates up to 188.4% for RS, 236.7% for wheat straw WS, 222.4% for LC and 131.1% for sugar beet pulp (SBP). Supplemental this combination enhanced the dry matter digestion (DMD), gas, NH 3 -N and VFA production, and MCP synthesis during in vitro rumen fermentation., Conclusions: The TrepCel3 and TrepCel4 exhibited the synergistic relationship (60:40) and significantly increased the saccharification yield of lignocellulosic substrates. The combination of them stimulated in vitro rumen fermentation of lignocellulosic substrates. This combination has the potential to be a feed additive to improve agricultural residues utilization in ruminants. If possible, in the future, experiments in vivo should be carried out to fully evaluate its effect., (© 2023. The Author(s).)

This study aimed to evaluate the effects of high-voltage pulsed electric fields (HPEF) and transglutaminase (TGase) cross-clinking on the physicochemical and rheological properties of Pleurotus eryngii protein (PEP). The results showed that HPEF increased α-helixes and β-turns but decreased β-folds. A HPEF at 1500 V/cm maximized the free sulfhydryl content and solubility of PEP. TGase formed high-molecular-weight polymers in PEP. TGase at 0.25% maximized the free sulfhydryl groups, particle size, and solubility; shifted the maximum absorption wavelength from 343 nm to 339 nm and 341 nm; increased α-helixes and β-turns and decreased β-folds; and showed better rheological properties. Compared with TGase cross-linking, HPEF-1500 V/cm and 1% TGase significantly reduced the free sulfhydryl groups, particle size, and solubility, produced more uniform network structures, and improved the rheological properties. These results suggest that HPEF can increase the cross-linking of TGase and improve rheological properties of TGase-cross-linked PEP by affecting the physicochemical properties.

Swainsonine induced liver inflammation in livestock; however, the underlying mechanisms, especially the role of bile acids (BAs), in the pathogenesis remained elusive. Here, our results showed that swainsonine induced hepatic inflammation via changing BA metabolism and gut microbiota in mice. Swainsonine significantly upregulated the levels of deoxycholic acid (DCA) and taurine-β-muricholic acid (T-β-MCA) in the serum and liver of mice due to the markedly increased genus Clostridium and the decreased genus Lactobacillus in the gut. As antagonists of the farnesoid X receptor (FXR), elevated DCA and T-β-MCA inhibited hepatic Fxr gene expression and thus suppressed FXR-SHP signaling and activated hepatic Cyp7a1 gene expression, which induced a significant upregulation of the total BA level in serum, contributing to liver inflammation. These findings offer new insights into the underlying mechanisms in which swainsonine induced liver inflammation in mice via the gut-liver axis and suggest that gut microbiota and its metabolite BAs may be underlying triggering factors.

To efficiently use lignocellulosic materials in ruminants, it is crucial to explore effective enzymes, especially bifunctional enzymes. In this study, a novel stable bifunctional cellulase-xylanase protein from buffalo rumen metagenome was expressed and characterized, CelXyn2. The enzyme displayed optimal activity at pH 6.0 and 45 °C. The residual endoglucanase and xylanase activities were 90.6% and 86.4% after a 60-min pre-incubation at 55 °C. Hydrolysis of rice straw, wheat straw, sheepgrass and sugar beet pulp by CelXyn2 showed its ability to degrade both cellulose and hemicellulose polymers. Treatment with CelXyn2 improved the hydrolysis of agricultural residues with an evident increase in production of total gas, lactate and volatile fatty acids. The results of 16S rRNA and real-time PCR showed that the effect on in vitro ruminal microbial community depended on fermentation substrates. This study demonstrated that CelXyn2 could strengthen lignocellulose hydrolysis and in vitro ruminal fermentation. These characteristics of CelXyn2 distinguish it as a promising candidate for agricultural application., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2023 The Authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd.)

Lian-Zhi-Fan (LZF) decoction is a hospital-prescribed traditional Chinese medicine botanical drug prepared by the fermentation of decocted Coptidis Rhizome (Huanglian), Gardeniae Fructus (Zhizi), and alum (Baifan). It has been used clinically in China for the treatment of anal fistula, perianal abscess, ulcerative colitis (UC), and other anorectal diseases for hundreds of years. However, due to the complexity of traditional Chinese medicine, the potential mechanisms of LZF in the treatment of UC have remained unknown. This study primarily investigated the remarkable pharmacological effects of LZF on TNBS-induced UC rats. To explore the complex targets and regulatory mechanisms of metabolic networks under LZF intervention, a metabolomics approach mediated by HPLC/Q-TOF-MS analysis was used to screen the different metabolites and their metabolic pathways in the serum in order to characterize the possible anti-UC mechanisms of LZF. After rectal administration of LZF for seven consecutive days, significant amelioration effects on body weight loss, DAI score, and colon inflammation were found in UC rats. Based on this, further metabolomics identified 14 potential biomarkers in the treatment of UC with LZF, of which five possessed diagnostic significance: L-alanine, taurocholic acid, niacinamide, cholic acid, and L-valine. These metabolites are mainly involved in 12 metabolic pathways, including nicotate and nicotinamide metabolism, glycospholipid metabolism, arginine and proline metabolism, primary bile acid biosynthesis, and pantothenate and CoA biosynthesis. These metabolic pathways suggest that LZF ameliorates UC by regulating amino acid metabolism, fat metabolism, and energy production. This study provides a useful approach for exploring the potential mechanisms of herbal prescription in UC treatment mediated by metabolomics., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Bai, Xiong, Wang, Cheng, Luo, Yang and Fu.)

Research Background: Sparassis latifolia polysaccharides can regulate lipids and cholesterol in serum and liver. However, little is known about the regulation mechanism of the polysaccharides on cholesterol metabolism and especially the causal relationship with gut microbiota regulation. This study will provide a theoretical basis for the cholesterol-lowering mechanism of S. latifolia polysaccharides and further development of functional foods., Experimental Approach: In this study, we investigated how the regulation mechanism of Sparassis latifolia polysaccharides affects intestinal cholesterol metabolism in high-fat and high-cholesterol diet-fed rats. Briefly, enzymatic colorimetric microplate assay was used to determine the concentration of faecal bile acid. Gas chromatography-mass spectrometry was used to detect the content of cholesterol and alcohol in faeces. Haematoxylin and eosin staining method was applied to observe the changes in the structure of the small intestine tissue. The related gene expressions in jejunum and ileum were detected by real-time fluorescent quantitative polymerase chain reaction. The related protein expressions in jejunum were studied by using Western blot. High-throughput sequencing was used to detect the intestinal flora changes of the caecal contents. Gas chromatography-mass spectrometry was applied to detect the concentration of short-chain fatty acids in the caecal content., Results and Conclusions: The results showed that Sparassis latifolia polysaccharides could improve the intestinal morphological structure and physiological indices in rats fed high-fat and high-cholesterol diet. Moreover, it could improve intestinal cholesterol metabolism disorder induced by high-fat and high-cholesterol diets via the reduction of the expression of HMGCR, NPC1L1, ACAT2, MTP, ASBT and IBABP mRNA or protein, increasing ABCG8 mRNA expression. In addition, it could also increase the relative abundance of Bacteroides , Butyricicoccus , Parabacteroides , Parasutteerella and Alloprevotella and the short-chain fatty acid concentration, to comprehensively regulate the intestinal cholesterol metabolism. The metabolomics analysis found that Sparassis latifolia polysaccharides could affect lipid, carbohydrate and other related metabolites. Some biomarkers associated with cholesterol metabolism correlated significantly with the abundance of specific intestinal microbiota., Novelty and Scientific Contribution: These findings indicate that Sparassis latifolia polysaccharides could attenuate intestinal cholesterol metabolism disorder, correlating with modulating gut microbiota and improving host metabolism. They provide theoretical support for the development of Sparassis latifolia as a new food resource., Competing Interests: CONFLICT OF INTEREST We declare that we have no financial or personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the position presented in, or the review of, this manuscript.

Feruloyl esterase (FAE; EC 3.1.1.73) cleaves the ester bond between ferulic acid (FA) and sugar, to assist the release of FAs and degradation of plant cell walls. In this study, two FAEs (Fae13961 and Fae16537) from the anaerobic fungus Pecoramyces sp. F1 were heterologously expressed in Pichia pastoris (P. pastoris). Compared with Fae16537, Fae13961 had higher catalytic efficiency. The optimum temperature and pH of both the FAEs were 45 ℃ and 7.0, respectively. They showed good stability-Fae16537 retained up to 80% activity after incubation at 37 ℃ for 24 h. The FAEs activity was enhanced by Ca 2+ and reduced by Zn 2+ , Mn 2+ , Fe 2+ and Fe 3+ . Additionally, the effect of FAEs on the hydrolytic efficiency of xylanase and cellulase was also determined. The FAE Fae13961 had synergistic effect with xylanase and it promoted the degradation of xylan substrates by xylanase, but it did not affect the degradation of cellulose substrates by cellulase. When Fae13961 was added in a mixture of xylanase and cellulase to degrade complex agricultural biomass, it significantly enhanced the mixture's ability to disintegrate complex substrates. These FAEs could serve as superior auxiliary enzymes for other lignocellulosic enzymes in the process of degradation of agricultural residues for industrial applications., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Background: As one of the most classic antineoplastic agents, doxorubicin (Dox) is extensively used to treat a wide range of cancers. Nevertheless, the clinical outcomes of Dox-based therapies are severely hampered due to the significant cardiotoxicity. Glycyrrhetinic acid (GA) is the major biologically active compound of licorice, one of the most well-known food additives and medicinal plants in the world. We previously demonstrated that GA has the potential capability to protect mice from Dox-induced cardiac injuries. However, the underlying cardioprotective mechanism remains unexplored., Purpose: To investigate the cardioprotective benefits of GA against Dox-induced cardiotoxicity and to elucidate its mechanisms of action., Study Design/methods: H9c2 cardiomyoblasts and AC16 cardiomyocytes were used as the cell models in vitro. A transgenic zebrafish model and a 4T1 mouse breast cancer model were applied to explore the cardioprotective effects of GA in vivo., Results: In vitro, GA inhibited Dox-induced cell death and LDH release in H9c2 and AC16 cells without affecting the anti-cancer effects of Dox. GA significantly alleviated Dox-induced ROS generation, mitochondrial dysfunction, and apoptosis in H9c2 cells. Moreover, GA abolished the expression of pro-apoptotic proteins and restored Nrf2/HO-1 signaling pathway in Dox-treated H9c2 cells. On the contrary, Nrf2 knockdown strongly abrogated the cardioprotective effects of GA on Dox-treated H9c2 cells. In vivo, GA attenuated Dox-induced cardiac dysfunction by restoring stroke volume, cardiac output, and fractional shortening in the transgenic zebrafish embryos. In a 4T1 mouse breast cancer model, GA dramatically prevented body weight loss, attenuated cardiac dysfunction, and prolonged survival rate in Dox-treated mice, without compromising Dox's anti-tumor efficacy. Consistently, GA attenuated oxidative injury, reduced cardiomyocytes apoptosis, and restored the expressions of Nrf2 and HO-1 in Dox-treated mouse hearts., Conclusion: GA protects against Dox-induced cardiotoxicity by suppressing oxidative stress, mitochondrial dysfunction, and apoptosis via upregulating Nrf2/HO-1 signaling pathway. These findings could provide solid evidence to support the further development of GA as a feasible and safe adjuvant to Dox chemotherapy for overcoming Dox-induced cardiotoxicity., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Increasing evidence indicates that type 2 diabetes mellitus (T2DM) is closely related to intestinal bacteria disorders and abnormal hepatic metabolism. Morchella importuna polysaccharide (MIP) shows excellent hypoglycemic activity in vitro. However, the hypoglycemic effect and mechanism of MIP in vivo have yet to be investigated. In this study, the blood glucose, blood lipid and insulin resistance of diabetic mice after MIP intervention were measured to evaluate its hypoglycemic effect. Then, the microbiome and metabolomics were combined to explore the hypoglycemic mechanism of MIP. Results indicated that high dose MIP (400 mg/kg) had significant hypoglycemic effect. Furthermore, MIP could reverse diabetes-induced intestinal disorder by increasing the abundance of Akkermansia, Blautia, Dubosiella, and Lachnospiraceae, as well as decreasing the abundance of Helicobacteraceae. Besides, the hepatic metabolites and complex network systems formed by multiple metabolic pathways were regulated after MIP treatment. Notably, a new biomarker of diabetes (N-P-coumaroyl spermidine) was discovered in this study. Moreover, the significant association between intestinal bacteria and hepatic metabolites was determined by correlations analysis, which in turn confirmed MIP alleviated T2DM via the gut-liver axis. Therefore, these findings elucidated in-depth hypoglycemic mechanisms of MIP and provided a new biomarker for the prevention of diabetes., Competing Interests: Declaration of competing interest There are no conflicts of interest to declare., (Copyright © 2022. Published by Elsevier B.V.)

To explore a scientific and reasonable cooking method for Clitocybe squamulosa , four cooking methods (boiling, steaming, microwaving, and frying) were applied to C. squamulosa , and the effects of different cooking methods on volatile flavor compounds, nutritional constituents, and antioxidant activities in C. squamulosa were systematically investigated. The results showed that 54, 53, 61, 63, and 49 volatile flavor compounds were detected in raw, boiled, steamed, microwaved, and fried samples, respectively. Large differences in volatile flavor compounds between the four cooking and raw samples were determined by using relative odor activity values (ROAV) and principal component analysis (PCA). In addition, steaming and microwaving could protect the nutrients of C. squamulosa , reduce losses during the cooking process and improve the color of cooked products compared to boiling and frying cooking methods. Meanwhile, cooking treatment exerted different effects on the antioxidant activity of C. squamulosa , and the antioxidant activity of C. squamulosa was the highest after microwave cooking treatment. This research can provide a theoretical basis for the cooking, processing and utilization of C. squamulosa and other wild edible fungi., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Yuan, Xu, Chang, Meng, Feng, Geng, Cheng and Liu.)

Yaks living on the Qinghai-Tibetan Plateau for a long time have evolved a series of mechanisms to adapt to the unique geographical environment and climate characteristics of the plateau. Compared with other ruminants, yaks have higher energy utilization and metabolic efficiency. This paper presents possible mechanisms responsible for the efficient energy utilization, absorption and metabolism resulting from the unique evolutionary process of yaks. It is hoped that the information discussed in this review will give a better insight into the uniqueness and superiority of yaks in regards to energy metabolism and utilization compared with cattle and open new avenues for the targeted regulation of energy utilization pathways of other ruminants., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, and there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2022 The Authors. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co. Ltd.)

This study examined the effect of the ratio of dietary metabolizable energy (MJ) to nitrogen (g) content (ME:N) on average daily gain (ADG), blood biochemical indices, rumen fermentation parameters, and rumen bacterial community in yaks. Thirty-six male yaks, aged 2-3 years, were divided into three groups and received a ME:N ratio of 0.42 (HY), 0.36 (MY,) or 0.32 (LY) MJ/g. Dry matter intake ranged between 3.16 and 3.63 kg/d and was lesser ( p  < 0.001) in the LY group than the other two groups. ME intake increased ( p  < 0.001) with an increase in the ME:N ratio, while N intake did not differ among groups. The ADG was 660 g/day for the MY group, which was higher ( p  < 0.005) than the 430 g/day in the LY group, while the HY group gained 560 g/day and did not differ from the other two groups. Feed intake to ADG ratio ranged between 5.95 and 7.95, and numerically was highest in the LY group and lowest in the MY group. In general, the concentration of ruminal total volatile fatty acids ( p  < 0.03) and molar proportions of propionate ( p  < 0.04), increased, while the molar proportion of acetate ( p  < 0.005) and the acetate:propionate ratio decreased ( p  < 0.001) with a decrease in the ME:N ratio. The molar proportion of butyrate did not differ among groups ( p  = 0.112). Group MY had higher ruminal NH 3 -N content than group HY and had a higher serum glucose content but lower urea content, lactate dehydrogenase, and creatine kinase content than group LY. In ruminal bacteria at the phylum level, the relative abundance of Firmicutes (F) was greater and of Bacteroidetes (B) was lesser, while the F:B ratio was greater in group MY than in groups HY an LY. We concluded that the yaks consuming the diet containing a ME:N ratio of 0.36 MJ/g had the best performance of the three groups., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Liu, Li, Hao, Degen, Wang, Ma, Niu, Cheng and Liu.)

α-galactosidase (EC 3.2.1.22) are glycosidases that catalyze the hydrolysis of α-1,6-linked D-galactosyl residues of different substrates, which has been widely applied in the food industry. Oudemansiella radicata is a kind of precious edible medicinal mushroom, which is a healthy, green, and safe food-derived enzyme source. In this study, a novel acidic α-galactosidase was purified from the dry fruiting bodies of O. radicata by ion-exchange chromatography and gel filtration, and designated as ORG ( O. radicata α-galactosidase). ORG was further immobilized to obtain iORG by the sodium alginate-chitosan co-immobilization method. Then, the characterization of free and immobilized enzymes and their potential application in the removal of the RFOs from soymilk were investigated. The results showed that ORG might be a 74 kDa heterodimer, and it exhibited maximum activity at 50 °C and pH 3.0, whereas iORG showed maximum activity at 50 °C and pH 5.5. In addition, iORG exhibited higher thermal stability, pH stability, storage stability, and a better degradation effect on raffinose family oligosaccharides (RFOs) in soymilk than ORG, and iORG completely hydrolyzed RFOs in soymilk at 50 °C within 3 h. Therefore, iORG might be a promising candidate in the food industry due to its excellent stability, high removal efficiency of RFOs from soymilk, and great reusability.

Background: Sparassis latifolia ( S. latifolia ) is a precious edible fungus with multiple biological activities. To date, no study has been investigated the underlying molecular mechanism of immunoregulation caused by the neutral polysaccharide of S. latifolia., Materials and Methods: To investigate immunomodulatory mechanism of S. latifolia neutral polysaccharide (SLNP), SLNP was obtained from S. latifolia and its structure, immune receptors and regulation mechanism were studied., Results: S. latifolia neutral polysaccharide consisted of arabinose, galactose, glucose, xylose, and mannose with a molar ratio of 6:12:63:10:5. SLNP was a pyran polysaccharide with a relative molecular weight of 3.2 × 10 5 Da. SLNP promoted the proliferation of RAW264.7, which further induced the secretions of nitric oxide, TNF-α, IL-6, and IFN-β, and upregulated the immune receptor TLR4 expression. Moreover, SLNP increased remarkably the levels of TRAF6, IRF3, JNK, ERK, p38, and p38 mRNA and protein mediated by TLR4., Conclusion: S. latifolia neutral polysaccharide regulated the immune function of RAW264.7 through MyD88-dependent and -independent signaling pathways mediated by TLR4 receptor, which suggests that SLNP is a new immunomodulator., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Qiao, Zhao, Wang, Cao, Chang, Yun, Cheng, Cheng and Feng.)

Although a natural anti-inflammatory ingredient, isoliquiritigenin (ISL), plays an effective role in ulcerative colitis (UC) treatment, a series of drawbacks still limit its clinical application, including the poor solubility, instability in gastrointestinal tract, and rapid elimination rate of ISL. Zein-based NPs display the benefits on drug loading and delivery, whereas with the poor stability. In this study, an edible nano-system composed by zein/caseinate complex was fabricated for the colon-targeting delivery of ISL, to improve its colon retention and anti-UC effects. The optimized ISL loaded zein/caseinate NPs (ISL@NPs) were prepared by single-factor design by anti-solvent precipitation method, and then characterized. The improved cellular uptake of ISL@NPs on NCM460 and RAW 264.7 cells was evaluated in vitro . The colon tissue permeability and retention capacity in vivo , and the anti-UC efficacy of ISL@NPs in DSS-induce UC were implemented. As a result, ISL@NPs with the high drug loading efficiency of 9.39% ± 0.26%, the average particle diameter of 137.32 ± 2.54 nm, exhibited the pH-sensitive stability in the different simulated gastrointestinal buffer. Compared with free ISL, ISL@NPs showed significantly higher cellular uptake ability in NCM460 and RAW 264.7 cells. Based on in vivo imaging system, zein/caseinate NPs showed the prolonged colonic retention and the enhanced penetration into the colonic epithelium. Finally, the oral administration of ISL@NPs could effectively alleviate the UC-related symptoms, down-regulate the production of pro-inflammatory factors, and reduce the infiltration of macrophages and neutrophils in colon tissues. In this study, an oral colon-specific nano-system, composed with the natural compound and edible materials, was developed as the promising alternatives in the prevention and treatment of UC., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Xiao, Wu, Cheng, Ma, Luo, Chang, Zhang, Chen, Zou, You and Zhang.)

Background: The limited therapeutic outcomes of atherosclerosis (AS) have allowed, traditional Chinese medicine has been well established as an alternative approach in ameliorating AS and associated clinical syndromes. Clinically, Tongsaimai tablet (TSMT), a commercial Chinese patent medicine approved by CFDA, shows an obvious therapeutic effect on AS treatment. However, its effective mechanism and quality control still need thorough and urgent exploration., Methods: The mice were orally administered with TSMT and their serum was investigated for the absorbed compounds using serum pharmacochemistry via the UPLC-Q-Exactive Orbitrap/MS analysis was employed to investigate these absorbed compounds in serum of mice orally administrated with TSMT. Based on these absorbed prototype compounds in serum derived from TSMT, a component-target-disease network was constructed using network pharmacology strategy, which elucidated the potential bioactive components, effective targets, and molecular mechanisms of TSMT against AS. Further, the screened compounds from the component-target network were utilized as the quality control (QC) markers, determining multi-component content determination and HPLC fingerprint to assess quality of nine batches of TSMT samples., Results: A total of 164 individual components were identified in TSMT. Among them, 29 prototype compounds were found in serum of mice administrated with TSMT. Based on these candidate prototype components, 34 protein targets and 151 pathways related to AS were predicted, and they might significantly exhibit potential anti-AS mechanisms via synergistic regulations of lipid regulation, shear stress, and anti-inflammation, etc. Five potentially bioactive ingredients in TSMT, including Ferulic acid, Liquiritin, Senkyunolide I, Luteolin and Glycyrrhizic acid in quantity not less than 1.2798, 0.4716, 0.5419, 0.1349, 4.0386 mg/g, respectively, screened from the component-target-pathway network. Thereby, these indicated that these five compounds of TMST which played vital roles in the attenuation of AS could serve as crucial marker compounds for quality control., Conclusions: Overall, based on the combination of serum pharmacochemistry and network pharmacology, the present study firstly provided a useful strategy to establish a quality assessment approach for TSMT by screening out the potential anti-AS mechanisms and chemical quality markers., (© 2022. The Author(s).)

Although the enrichment of antibiotic resistance genes (ARGs) in diverse organic soils have been explored, understanding of the ecological processes governing the composition of ARGs in long-term organically fertilized soils still remains limited across typical agricultural regions. Thus, the distribution and assembly of ARG profile in three typical agricultural soils (black soil, fluvo-aquic soil, and red soil) under long-term contrasting fertilization regimes (chemical-only vs organic-only) were investigated using high-throughput qPCR (HT-qPCR). The application of organic manure significantly increased the abundance and number of ARGs across soils, as compared to those with chemical fertilizer. Organic manure application enriched the abundance of mobile genetic elements (MGEs), which were positively associated with ARGs. In addition, it is long-term organic fertilizer that enriched the number and abundance of opportunist and specialist ARGs in the fluvo-aquic and red soils, but not black soils. The number and abundance of most generalist ARGs did not change significantly among different fertilization or soil types. The assembly process of the ARG profiles tends to be more deterministic in organically fertilized soils than in chemically fertilized soils. These results suggest that long-term organic fertilizer application may contribute to the persistence and health risk of the soil antibiotic resistomes (especially specialist ARGs)., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This study aimed to investigate the changes in the blood metabolic profiles of grazing yaks during the cold season to reveal their physiological status and seek the nutrients needed to be supplemented. Six castrated yaks (3 years old) with 166.8 kg (standard deviation = 5.3) of liveweight grazed in the Qinghai-Tibetan Plateau were used as experimental animals without supplementary feeding. Blood samples of each animal were collected in October and December 2015, and March 2016 for the analysis of serum biochemicals and metabolome. Results showed serum indices involved in protein metabolism in grazing yaks showed greater differences during the cold season than the metabolisms of energy or minerals. Cold stress in December had minor effects on the serum metabolic profiles of yaks compared with those in October. Yaks in October and December shared seven differential serum metabolites and enrichments of the "arachidonic acid metabolism" and "glycine, serine, and threonine metabolism" pathways compared with those in March caused by the shortage of feeds. Summarily, the nutrient deficiency would be influential on the physiological status of grazing yaks during the cold season, especially on the protein metabolism, which could be improved by supplementary feeds.

Phytosterols are natural steroids in plants, possessing bioactivities that could modify gut microbes. This experiment aimed to evaluate the effects of feeding phytosterols on the community structures and metabolic functions of the rumen microbiota in perinatal cows. Perinatal cows were supplied with 0 mg (control) or 200 mg (treatment) phytosterols per day. Multiomic analyses were used to analyze the community structures and metabolic functions of rumen microbiota. Results showed that dietary phytosterols increased the copy number of total ruminal bacteria, the concentration of microbial crude protein, and the molar percentage of propionate in the rumen of perinatal cows but had no effects on the alpha diversity of ruminal bacteria. However, they enriched three genera (i.e., Fibrobacter ) and seven species (i.e., Fibrobacter succinogenes) within active ruminal bacteria. Metatranscriptomic and metabolomic analyses revealed that dietary phytosterols enhanced the pathway of glycolysis and the family of glycoside hydrolase 13 but depressed the citrate cycle and pyruvate metabolism and several pathways of amino acid biosynthesis. In conclusion, dietary addition of phytosterols improved the growth of ruminal bacteria and changed rumen fermentation by modifying the rumen microbiome and the energy metabolism pathways, which would be beneficial for the energy utilization of perinatal cows. IMPORTANCE Perinatal cows suffer serious physiological stress and energy deficiency. Phytosterols have bioactive functions for gut microbes. However, little knowledge is available on their effects on rumen microbiota and rumen fermentation. Results of the present experiment revealed that dietary supplementation of phytosterols could improve the growth of ruminal bacteria and changed the rumen fermentation to provide more glycogenetic precursors for the perinatal cows by modifying the ruminal bacteria community and altering the energy metabolism pathways of the rumen microbiota. These findings suggest that dietary supplementation of phytosterols would be beneficial for perinatal cows suffering from a negative energy balance.