Water-in-oil Pickering emulsion using ergosterol as an emulsifier solely.

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.

Self-Amplified pH/ROS Dual-Responsive Co-Delivery Nano-System with Chemo-Photodynamic Combination Therapy in Hepatic Carcinoma Treatment.

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.

Dietary supplementing phytosterols improves the metabolic status of perinatal cows revealed by plasma metabolomics and faecal microbial metabolism.

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 ß-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.

Influence of dietary Salicornia europaea L. extract supplementation on feed efficiency of Altay sheep by modifying their gastrointestinal bacteria communities.

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.

Effect of Different Drying Methods on the Quality of Oudemansiella raphanipes.

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.

The effect of sequential extraction on the physicochemical and rheological properties of Naematelia aurantialba polysaccharides.

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.

Methane production and lignocellulosic degradation of wastes from rice, corn and sugarcane by natural anaerobic fungi-methanogens co-culture.

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.

Dynamic changes of rumen microbiota and serum metabolome revealed increases in meat quality and growth performances of sheep fed bio-fermented rice straw.

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.

The Effects of Naematelia aurantialba on the Pasting and Rheological Properties of Starch and the Research and Development of Soft Candy.

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.

Polysaccharide from Clitocybe squamulosa: Gel-forming properties and its compound effect with food thickener.

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 Ca2+ 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.

Effects of different extraction methods on the physico-chemical characteristics and biological activities of polysaccharides from Clitocybe squamulosa.

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 (IC50 = 0.29) and ABTS free radicals (IC50 = 0.19), total reduction ability (IC50 = 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.

A Comparative Study of Growth Performance, Blood Biochemistry, Rumen Fermentation, and Ruminal and Fecal Bacterial Structure between Yaks and Cattle Raised under High Concentrate Feeding Conditions.

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.

[Q-marker prediction of resin ethanol extract of Gegen Qinlian Decoction based on characteristic spectrum and network pharmacology].

The resin ethanol extract of Gegen Qinlian Decoction(GGQLD) has been found to significantly alleviate the intestinal toxicity caused by Irinotecan, but further research is needed to establish its overall quality and clinical medication standards. This study aimed to establish an HPLC characteristic fingerprint of the resin ethanol extract of GGQLD, predicted the targets and signaling pathways of its pharmacological effects based on network pharmacology, identified core compounds with pharmacological relevance, and analyzed potential quality markers(Q-markers) of the resin eluate of GGQLD for relieving Irinotecan-induced toxicity. By considering the uniqueness, measurability, and traceability of Q-markers based on the "five principles" of Q-markers and combining them with network pharmacology techniques, the overall efficacy of the resin ethanol extract of GGQLD can be characterized. Preliminary predictions suggested that the four components of puerarin, berberine, baicalin, and baicalein might serve as potential Q-markers for the resin etha-nol extract of GGQLD. This study provides a basis and references for the quality control and clinical mechanism of the resin ethanol extract of GGQLD.

Polysaccharide from Sparassis latifolia alleviates intestinal barrier dysfunction in mice exposed to lead.

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.

Effects of Freeze-Thaw Cycles on the Structures and Functional Properties of Clitocybe squamulosa Protein Isolates.

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.

Garcinone E suppresses breast cancer growth and metastasis by modulating tumor-associated macrophages polarization via STAT6 signaling.

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.

[Equivalence of combined decoction and mixed single decoctions of Gegen Qinlian Decoction in alleviating chemotherapy-associated diarrhea].

This study compared the chemical profiles, component content, dry paste yield, and pharmacological effects of samples obtained from the mixed single decoctions and the combined decoction of Gegen Qinlian Decoction(GQD), aiming to provide an experimental foundation for evaluating the equivalence of the two decocting methods and the suitability of TCM formula granules in clinical application. The same decoction process was used to prepare the combined decoction and mixed single decoctions of GQD. Ultra-performance liquid chromatography coupled with Q-Exactive Orbitrap mass spectrometry(UPLC-Q-Exactive Orbitrap MS) was employed to compare the chemical profiles between the two groups. High-performance liquid chromatography(HPLC) was used to compare the content of nine characteristic components between the two groups. Then, a delayed diarrhea mouse model induced by irinotecan was established to compare the pharmacological effects of the two groups on chemotherapy-induced diarrhea. The UPLC-Q-Exactive Orbitrap MS in ESI~+ and ESI~- modes identified 59 chemical components in the compound decoction and mixed single decoctions, which showed no obvious differences in component species. The content of baicalin and wogonoside was higher in the compound decoction, while that of puerarin, daidzein-8-C-apiosylglucoside, berberine, epiberberine, wogonin, glycyrrhizic acid, and daidzein was higher in the mixed single decoctions. Further statistical analysis revealed no significant difference in the content of the nine characteristic components between the compound decoction and the mixed single decoctions. The dry paste yield had no significant difference between the two groups. Compared with the model group, both compound decoction and mixed single decoctions alleviated the weight loss and reduced diarrhea index in mice. Both of them lowered the levels of tumor necrosis factor-α(TNF-α), interleukin-1ß(IL-1ß), cyclooxygenase-2(COX-2), intercellular adhesion molecule-1(ICAM-1), interleukin-10(IL-10), malondialdehyde(MDA), and nitric oxide(NO) in the colon tissue. Furthermore, they significantly increased the levels of glutathione peroxidase(GSH-Px) and superoxide dismutase(SOD). Hematoxylin-eosin(HE) staining showed that colon tissue cells were tightly arranged with clear nuclei in both groups without obvious difference. The compound decoction and mixed single decoctions showed no significant differences in chemical component species, content of nine characteristic components, dry paste yield, or the pharmacological effects on alleviating chemotherapy-induced diarrhea. The findings provide a reference for evaluating the flexibility and superiority of combined or single decocting method in the preparation of TCM decoctions or formula granules.

Transglutaminase-Catalyzed Glycosylation Improved Physicochemical and Functional Properties of Lentinus edodes Protein Fraction.

Lentinula edodes has high nutritional value and abundant protein. In order to develop and utilize edible mushroom protein, this study was designed to investigate the effects of TGase-catalyzed glycosylation and cross-linking on the physicochemical and functional properties of Lentinus edodes protein fraction. The results showed that within a certain time, glycosylation and TGase-catalyzed glycosylation decreased the total sulfydryl, free sulfydryl, disulfide bond, surface hydrophobicity, ß-fold and α-helix, but increased the fluorescence intensity, random coil, ß-turn, particle size and thermal stability. The apparent viscosity and the shear stress of the protein with an increase in shear rate were increased, indicating that TGase-catalyzed glycosylation promoted the generation of cross-linked polymers. In addition, the TGase-catalyzed glycosylated proteins showed a compact texture structure similar to the glycosylated proteins at the beginning, indicating that they formed a stable three-dimensional network structure. The flaky structure of proteins became more and more obvious with time. Moreover, the solubility, emulsification, stability and oil-holding capacity of enzymatic glycosylated Lentinus edodes protein fraction were significantly improved because of the proper TGase effects of glycosylation grafting and cross-linking. These results showed that glycosylation and TGase-catalyzed glycosylation could improve the processing characteristics of the Lentinula edodes protein fraction to varying degrees.

Characterization of a novel bifunctional enzyme from buffalo rumen metagenome and its effect on in vitro ruminal fermentation and microbial community composition.

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.

Glycyrrhetinic acid suppresses breast cancer metastasis by inhibiting M2-like macrophage polarization via activating JNK1/2 signaling.

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.