Phenolic Constituents from Black Quinoa Alleviate Insulin Resistance in HepG2 Cells via Regulating IRS1/PI3K/Akt/GLUTs Signaling Pathways.

Quinoa is a nutrient-rich pseudocereal with a lower glycemic index and glycemic load. However, its therapeutic potency and underlying mechanism against insulin resistance (IR) have not been fully elucidated. In this work, network pharmacology was applied to screen IR targets and their related pathways. The efficacy and mechanism of black quinoa polyphenols (BQP) on IR improvement were evaluated and uncovered based on the IR model in vitro combined with molecular docking. Ten phenolic constituents of BQP were detected, and the network pharmacology results show that PI3K/Akt pathways are the main pathways in BQP against IR. The in vitro assay proved that BQP increases the glucose consumption and glycogen synthesis via upregulating insulin receptor substrate 1 (IRS1)/PI3K/Akt/glucose transporters (GLUTs) signaling pathways to alleviate IR. Rutin, resveratrol, and catechin show lower binding energy docking with IRS1, PI3K, Akt, and GLUT4 proteins, indicating better interactions. It might be an effective constituent against IR. Hence, BQP could become a potential functional food source for blood glucose management among insulin-resistant people.

Novel hydrocortisone magnetic molecularly imprinted polymers: Preparation, characterization, and application.

In this work, magnetic molecularly imprinted polymers (MMIPs) were prepared by a surface imprinting method using Fe3O4 nanoparticles as a support. The products were characterized by FT-IR spectroscopy, VSM, TGA, SEM, and TEM. Combined with HPLC, hydrocortisone in milk powder and milk were separated and purified, and their contents were monitored. The results showed that MMIPs with a particle size of approximately 1000 nm were successfully prepared. The adsorption mechanism of MMIPs was confirmed by kinetic adsorption and thermodynamic adsorption experiments; the maximum adsorption capacity was found to be 17.2 mg g-1, and adsorption equilibrium could be reached within 40 min. In the actual sample application, the recovery rates of milk powder and milk were 93.88-99.15% and 95.80-98.10%, respectively. These results showed that MMIPs had good performance in selectively identifying hydrocortisone and were suitable for determining hydrocortisone in milk products.

Ginger polysaccharides enhance intestinal immunity by modulating gut microbiota in cyclophosphamide-induced immunosuppressed mice.

In this study, the immunity-enhancing effect of ginger polysaccharides UGP1 and UGP2 on CTX-induced immunosuppressed mice was evaluated. The results showed that ginger polysaccharide could effectively alleviate the symptoms of weight loss and dietary intake reduction induced by CTX, increase fecal water content, reduce fecal pH, and protect immune organs of immunosuppressed mice. In addition, ginger polysaccharides also stimulated the secretion of cytokines IL-2, IL-4, TNF-α and immunoglobulin Ig-G in the serum of mice, increased the expression of Occludin and Claudin-1, and restored the level of short-chain fatty acids in the intestine to improve immune deficiency. Furthermore, ginger polysaccharides significantly reduced the relative abundance ratio of the Firmicutes and Bacteroidetes in mice and increased the relative abundance of Verrucomicrobia and Bacteroidetes at the phylum level. At the family level, ginger polysaccharides increased the relative abundance of beneficial bacteria such as Muribaculaceae, Bacteroidaceae and Lactobacillaceae, and decreased the relative abundance of harmful bacteria such as Rikenellaceae and Lachnospiraceae. Spearman correlation analysis indicated that ginger polysaccharides could enhance intestinal immunity by modulating gut microbiota associated with immune function. These results indicated that ginger polysaccharides have the potential to be a functional food ingredients or a natural medicine for the treatment of intestinal barrier injury.

Antidiabetic activities of glycoprotein from pea (Pisum sativum L.) in STZ-induced diabetic mice.

Polysaccharides have hypoglycemic activity and pea protein has high nutritional value. The purified pea glycoprotein PGP2 has been shown to inhibit the activity of α-glucosidase and α-amylase in previous studies. To study the mechanism of PGP2-induced blood glucose lowering in vivo, this paper established a diabetic mouse model by intraperitoneal injection of STZ and high-fat diet, and evaluated the blood-glucose-lowering activity of the pea component PGP2 at different doses. The results showed that intragastric administration of PGP2 could effectively reduce diabetic weight loss and polyphagia symptoms, reduce fasting blood glucose levels in mice, and improve oral glucose tolerance levels in mice. PGP2 could promote insulin secretion and had a protective effect on mouse organs. After intragastric administration of PGP2 in mice, the serum levels of total cholesterol, triglycerides and low-density lipoprotein decreased. PGP2 up-regulated the gene expression of insulin receptor substrates IRS-1 and IRS-2 in liver tissues, thereby reducing insulin resistance. Based on the above experimental results, PGP2 had good hypoglycemic activity and was expected to be developed as a natural medicine for the treatment of type II diabetes.

A new type of magnetic molecular imprinted material combined with ß-cyclodextrin for the selective adsorption of zearalenone.

In this paper, a new magnetic molecular imprinted polymer-cyclodextrin (MMIP-CD) material was prepared by connecting ß-cyclodextrin (CD) on the surface of a magnetic molecular imprinted polymer (MMIP) and used for the rapid and specific adsorption of zearalenone (ZEN). By using warfarin as the virtual template molecule, tetraethyl orthosilicate (TEOS) as the crosslinking agent, and (3-aminopropyl) triethoxysilane (APTES) as the functional monomer, a MMIP was produced by surface imprinting technology. Sulfobutyl ether-ß-cyclodextrin attached to the surface of the MMIP under heating conditions produced a new specific adsorption material with exceptional adsorption capacity and excellent selectivity for ZEN. Scanning electron microscopy (SEM), transmission electron microscopy (TEM) and TEM-mapping results showed that the prepared MMIP-CD had a uniform particle size of about 480 nm, and the molecularly imprinted layer was successfully wrapped on the surface of the nanoparticles with a thickness of about 50 nm, whereby the cyclodextrin was effectively attached to the surface of the MMIP. The adsorption mechanism of MMIP-CD was confirmed by kinetic adsorption and thermodynamic adsorption experiments, the maximum adsorption capacity was found to be about 30 mg g-1, and the adsorption equilibrium could be reached within 20 min. The value of IF (QMMIP-CD/QMNIP) is 4.642. This showed that compared with MNIP, MMIP-CD showed a greatly improved specific adsorption capacity of ZEN. Selective experiments proved that MMIP-CD effectively combined the advantages of MMIP and CD, enhancing the adsorption capacity together with reducing the disadvantages that MMIP cannot distinguish structural analogs and CD cannot identify hydrophobic compounds effectively. In actual sample testing, the limit of quantification (LOQ) and limit of detection (LOD) were 0.1 ng kg-1 and 0.3 ng kg-1, respectively. The stability and detection precision of this method were 0.98-2.76% and 1.67-3.88%, respectively. The results proved that MMIP-CD had good development potential in the field of selective adsorption of ZEN, and laid the foundation for follow-up research.

Preparation of magnetic molecularly imprinted polymers for the identification of zearalenone in grains.

This study was based on the specific binding ability of magnetic molecularly imprinted polymers (MMIPs) combined with a high-performance liquid chromatography-fluorescence detector (HPLC-FLD) for the rapid determination of zearalenone (ZEN) in cereals. A novel magnetic molecularly imprinted polymer was prepared by surface imprinting technology. Warfarin was used as a virtual template, 3-aminopropyl triethoxysilane (APTES) was used as the functional monomer, and tetraethyl orthosilicate (TEOS) was used as the cross-linking agent. Analysis by a vibrating sample magnetometer (VSM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) showed that MMIPs were prepared with a particle size about 450 nm, the imprinted molecular layer accounting for 10.7% of the total mass, and saturation magnetization of about 34.54 emu/g. The maximum adsorption capacity (Qmax) of the thermodynamic and kinetic adsorption experiments were 13.90 mg/g and 8.71 mg/g, respectively. The Langmuir model showed that the binding sites were uniformly distributed on the surface of the MMIPs. The Scatchard analysis showed that MMIPs had two types of binding sites with Qmax of 8.22 mg/g and 15.37 mg/g, respectively. In actual sample detection, the limit of detection (LOD) and limit of quantification (LOQ) were 0.4 ng/kg and 0.9 ng/kg, respectively. The sample recovery rate was 90.56-99.96%, the daytime stability was 1.35-2.87%. These results showed that MMIPs had good performance in selectively identifying ZEN and were suitable for determining ZEN in cereals.

Effects of a ß-type glycosidic polysaccharide from Flammulina velutipes on anti-inflammation and gut microbiota modulation in colitis mice.

Using the Flammulina velutipes polysaccharide (FVP) extracted from our previous study, herein, we investigated the improvement of this starch-free ß-type glycosidic polysaccharide in alleviating dextran sodium sulfate-induced ulcerative colitis (UC) in mice. The absolute and relative abundance of intestinal microbes in the mouse feces were both determined by 16s RNA gene sequencing. The results from the histological analysis indicate that FVP treatment reduced the symptoms of UC, up- or down-regulated the relative gene expression levels in the colon tissue, and enhanced the capacity of metabolic and biogenesis in the UC mice, as predicted by PICRUSt. 11 species of gut microbes including Lactobacillus, Bifidobacterium, and Clostridium associated with UC symptoms were analyzed by correlation analysis. Our findings suggest that FVP can alleviate the UC symptoms in mice by regulating specific gut microbes, improving the understanding of the functional activity of FVPs as prebiotics.

Preparation of magnetic molecularly imprinted polymer for selective identification of patulin in juice.

A highly efficient and selective method was successfully developed by using magnetic molecularly imprinted polymers (MMIPs) combined with high performance liquid chromatography (HPLC) to quickly determine patulin (PAT) in juice. MMIPs was prepared by surface imprinting method using Fe3O4 nanoparticles as supporter, 2-oxindole as virtual template, (3-Aminopropyl) triethoxysilane (APTES) as functional monomer and tetraethyl orthosilicate (TEOS) as crosslinking agent. The structure of the product was characterized by vibrating sample magnetometer (VSM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). The results showed that MMIP with a particle size of about 450 nm was successfully prepared, the imprinted molecular layer accounted for about 11.6% of the total mass, and the saturation magnetization was about 6.82 emu/g. The maximum adsorption capacities (Qmax) of kinetic and thermodynamic adsorption experiments were 1.97 mg/g and 4.241 mg/g, respectively. The adsorption process was highly selective and fitted well with the pseudo-second-order model. Langmuir model demonstrated that the binding sites were evenly distributed on the surface of the MMIPs. Scatchard analysis showed that MMIPs had two types of binding sites with Qmax of 4.53 mg/g and 5.73 mg/g, respectively. In the actual sample application, the limit of detection (LOD) and the limit of quantification (LOQ) were 3 µg/kg and 10 µg/kg. And the recovery rate of the sample was 86.44-95.50%. MMIPs possessed excellent applicability with stability of 1.11-3.16% and accuracy of 0.63-1.94%. These results indicated that MMIPs had good performance in separating PAT and was suitable for determining PAT in actual samples.

Characterization and antitumor activities of polysaccharides obtained from ginger (Zingiber officinale) by different extraction methods.

Three different extraction technologies including hot water extraction (HWE), enzyme assisted extraction (EAE) and ultrasonic cell grinder extraction (UCGE) were employed to extract crude ginger polysaccharides (GPs) under their respective best parameters, then crude GPs were purified by DEAE cellulose-52 and Sephadex G-200 size-exclusion chromatography in that order. Five GPs fractions (HGP, EGP1, EGP2, UGP1, and UGP2, respectively) were obtained. The differences of five GPs in chemical composition, characterization and antitumor activities were further compared. The molecular weights were different in five GPs, varying from 11.81 to 1831.75 kDa. Mannose and glucose as the main monosaccharide and the glycosidic linkage of →4)-α-D-Glc(1→ and -α-Manp-(1→ existed in both five GPs. While EGP2 and UGP1 possessed specific structure of →6)-ß-D-Galp-(1→ and UGP1 contained more sulfate group. Moreover, UGP1 exhibited strong inhibitory effect on three tumor cells especially the colon cancer. The inhibition rates of UGP1 on H1975, HCT116 and MCF-7 were 23.339 ± 2.285%, 56.843 ± 2.405% and 21.061 ± 1.920% respectively. The study indicated GPs extracted by UCGE could reserve more active structure and inhibit colon cancer more significantly.

Impact of mushroom (Pleurotus eryngii) flour upon quality attributes of wheat dough and functional cookies-baked products.

The objective of this study was to create a healthier version of a commonly consumed baked food (cookies) by replacing some of the wheat flour with a nutraceutical-rich mushroom flour. The impact of incorporating different levels of powdered mushroom (Pleurotus eryngii) flour on the rheological properties of the cookie dough and the final cookies was therefore determined. The rheological properties of wheat dough supplemented with 0%-25% (w/w) of mushroom flour were analyzed using a Mixolab instrument and a shear rheometer. Increasing the ratio of mushroom-to-wheat flour in the doughs increased the peak and final viscosities, but decreased dough stability and elastic modulus. Sensory evaluation using an orthogonal test showed that 15% mushroom flour, 10% maltodextrin, and 1.5% sodium bicarbonate were the optimal composition for producing cookies with the best sensory score. In conclusion, our results showed that cookies with acceptable textures and appearances could be produced by replacing up to 15% of wheat flour with mushroom flour.

Purification, characterization and anti-tumor activities of polysaccharides from Ecklonia kurome obtained by three different extraction methods.

To investigate and compare the effects of different extraction methods on the structure and anti-tumor activity of Ecklonia kurome polysaccharides (EP), three techniques, namely hot water extraction (HW), ultrasonic-assisted extraction (UA) and enzyme-assisted extraction (EA), were used to extract EP, and three crude EPs were purified by DEAE-cellulose and gel filtration chromatography. The significant antitumor active components in each method were screened by MTT assay and named as HW-EP5, UA-EP4 and EA-EP3, respectively. The molecular weight, FT-IR assay and NMR showed that HW-EP5, UA-EP4 and EA-EP3 were pyran polysaccharides with a molecular weight of 14,466, 15,922 and 16,947 Da, respectively. HW-EP5 contained the most monosaccharides and the highest content of sulfate and uronic acid. HW-EP5 had an even and smooth sheet-like appearance, while UA-EP4 and EA-EP3 exhibited irregular and rough fragments. All three polysaccharides can inhibit the migration of human breast cancer cells (MCF-7) and promote its apoptosis. All three polysaccharides promoted caspase activity during apoptosis. HW-EP5 and UA-EP4 up-regulated the expression of proapoptotic proteins Bax and p53, while EA-EP3 only up-regulated the expression of p53. These experimental results indicate that Ecklonia kurome polysaccharides, especially HW-EP5, have great potential as a natural medicine for the treatment of breast cancer.

16α-Hydroxytrametenolic Acid from Poria cocos Improves Intestinal Barrier Function Through the Glucocorticoid Receptor-Mediated PI3K/Akt/NF-κB Pathway.

This study evaluated the effect of triterpenoids from edible mushroom Poria cocos on intestinal epithelium integrity and revealed the transcriptional regulatory pathways that underpin restorative mechanisms in the gut. Based on computational docking studies, transcriptional activation experiments and glucocorticoid receptor (GR) protein immunofluorescence localization assays in cultured cells, 16α-hydroxytrametenolic acid (HTA) was discovered as a novel GR agonist in this study. HTA ameliorates TNF-α-induced Caco-2 monolayer intestinal epithelial barrier damage and suppressed activation of phosphatidylinositol 3-kinase (PI3K) and protein kinase B (Akt), which attenuated downstream IκB and nuclear factor kappa-B (NF-κB) phosphorylation through GR activation. Moreover, HTA prevented NF-κB translocation into the nucleus and binding to its cis-element and suppressed lipopolysaccharide-induced downstream NO production and pro-inflammatory cytokines at both protein and mRNA expression levels. In conclusion, HTA from P. cocos improves intestinal barrier function through a GR-mediated PI3K/Akt/NF-κB signaling pathway and may be potentially exploited as a supportive dietary therapeutic strategy for restoring gut health.

Characterization and antioxidant activity of polysaccharides obtained from ginger pomace using two different extraction processes.

In this study, two different processes of hot water (HW) and ultrasonic-assisted (UA) for the extraction of polysaccharide from ginger pomace (GPPs) were employed under their respective best parameters, and the characterization and antioxidant activity of the purified polysaccharide (HW-GPP1, HW-GPP2, HW-GPP3, and UA-GPP1, UA-GPP2, UA-GPP3, respectively) were analyzed. The data implied that the yield of the polysaccharide obtained by UA was higher than that of HW. Meanwhile, two kinds of GPPs possessed the different preliminary structural characteristics including molecular weight distributions, total sugar and protein content, uronic acid content, while similar monosaccharide compositions and sulfuric radical contents. In vitro antioxidant activity assays indicated that UA-GPP3 showed the strongest scavenging abilities on DPPH radicals, while UA-GPP2 possessed the strongest scavenging abilities on hydroxyl and superoxide radicals. Moreover, the antioxidant activity of each fractions of GPPs extracted by UA was better than that of the corresponding fractions of GPPs extracted by HW. These results showed that UA was more beneficial to enhance the extraction yields of the polysaccharides, and also resulted in GPPs with higher bioactivity. Therefore, it indicated that UA-GPPs could be used as a potential natural antioxidant. Accordingly, the ginger pomace could be used as a potential source for natural antioxidant.

Effects of ultrasound-assisted extraction on antioxidant activity and bidirectional immunomodulatory activity of Flammulina velutipes polysaccharide.

In order to investigate the impacts of ultrasound-assisted extraction (UAE) on Flammulina velutipes polysaccharides (FVPs), the differences between FVPs extracted by ultrasound-assisted extraction (FVPU) and FVPs extracted by hot water extraction (FVPH) were compared in terms of yield, primary compositions, surface microstructure, helix-coil transition structure, molecular weight distribution, antioxidant activity, and bidirectional immunomodulatory activity. Results indicated that UAE changed the above properties of FVPs. Compared with FVPH, higher yield, protein content, and uronic acid content but lower polysaccharide and polyphenol contents were observed in FVPU. UAE changed the surface microstructure, destroyed the triple helix structure, and increased the proportion of low molecular weight polysaccharide components of FVPU. Compared with FVPH, FVPU showed a stronger reducing power and scavenging activities on DPPH radical, hydroxyl radical, and superoxide anion radical. FVPU was a better inhibitor of inflammation compared with FVPH. However, FVPH had a better immunity enhancing effect compared with FVPU. These results were attributed to the cavitation effect of ultrasonic waves on the structure of polysaccharides during the extraction process of UAE. These findings suggested that UAE was an efficient and environmentally friendly method to produce new polysaccharides from F. velutipes for the development of functional foods or nutraceuticals.

Preparation and in vitro absorption studies of a novel polysaccharide­iron (III) complex from Flammulina velutipes.

Iron is essential in fundamental bioactivities, so it makes sense to improve the efficiency of iron on epithelial transport. In this work, a novel polysaccharide­iron(III) complex (FVP-Fe(III)) was prepared from Flammulina velutipes with a specific structure. The FVP-Fe(III) had a molecular weight of 15.13 kDa with a monosaccharide composition of mannose, galacturonic acid, glucose, galactose, xylose, fucose in a molar ratio 3.6:2.1:60.8:18.7:3.9:10.9. In the vitro digestion model, the complex could maintain better solubility and steady of iron than FeSO4. In the cell assay, FVP-Fe(III) showed lower cytotoxicity and better absorption. The transport amount of FVP-Fe(III) was 1.5-fold of FeSO4 at same concentration and 1.8-fold of FeSO4 at same time. The transport was mediated by the peptide transporter (pepT1) active transport pathway and the efflux of the sample was mainly mediated by multidrug-resistance proteins (MRP) transporter. The results of this study suggested that the polysaccharide obtained from F. velutipes could be developed a new kind of iron delivery for further study.

Immunomodulatory activities of proteins from Astragalus membranaceus waste.

BACKGROUND: Astragalus membranaceus is a traditional Chinese medicine that has a long history of medical applications. It is of interest to investigate the functional components of A. membranaceus waste with regard to its development and utilization and increasing resource utilization. RESULTS: The protein AMWP was isolated from the A. membranaceus waste. This protein was further purified by DEAE-cellulose-52 chromatography and Sephadex G-200 size-exclusion chromatography to obtain three fractions, named AMWPDG2, AMWPDG4 and AMWPDG6. Then, their immunomodulatory activities were evaluated by using cell model experiments. The results indicated that the protein fractions could significantly increase the proliferation of splenic lymphocytes, peritoneal macrophages and bone-marrow-derived cells (BMDCs). AMWPDG2 showed the highest immunocompetence. AMWPDG2, AMWPDG4 and AMWPDG6 not only significantly improved the phagocytosis and immunomodulatory factors (interleukin (IL)-6, tumor necrosis factor-α, nitric oxide, hydrogen peroxide) secretion of peritoneal macrophages, but also promoted the expression of inflammatory cytokines (IL-6, IL-12 p40, IL-1ß, IL-1α) and chemokines (CXCL1, CCL3) in BMDCs. CONCLUSION: Taken together, these results indicated that three protein fractions from the A. membranaceus waste might be a potential natural immunomodulator. Moreover, it also provided the theoretical basis for further researching the mechanism of AMWPDG2, AMWPDG4 and AMWPDG6 on improving the immune response. © 2019 Society of Chemical Industry.

Identification of ortho-naphthoquinones as anti-AML agents by highly efficient oxidation of phenols.

A straightforward method for synthesizing ortho-naphthoquinones was identified using an easily available cobalt-Schiff base complex. Efficient oxidation of phenols to ortho-naphthoquinones was useful in obtaining compounds with potent biological activity for the treatment of acute myeloid leukemia (AML). Among these compounds, the compound 4h effectively inhibited the proliferation of different AML cell lines in vitro. Further in vivo antitumor studies indicated that 4h at 40 mg/kg/d led to tumor regression in led to tumor regression in an MV4-11 xenograft model without evident toxicity. The cobalt-Schiff base complex was found to be an efficient catalyst in the transformation of phenols to ortho-quinones, and the compound 4h represents a potential scaffold to optimize the production of a treatment for AML.

Simultaneous separation and determination of six arsenic species in Shiitake (Lentinus edodes) mushrooms: Method development and applications.

A method for simultaneously separating six arsenic (As) species was established by ultrasound-assisted extraction-anion exchange chromatography coupled with inductively coupled plasma mass spectrometry. Six As species could be well separated within 15 min when 0.3 M acetic acid and 25 mM NH4H2PO4 were used as the extraction solvent and mobile phase, respectively. Under optimized conditions, the limits of detection and recovery of As species and the relative standard deviation were in the ranges of 0.31-0.59 µg/L, 94.30-102.75%, and 1.63-3.72%, respectively. Furthermore, the established method was successfully applied to fresh Shiitake (Lentinus edodes) mushrooms and processed products. The inorganic As contents of all samples ranged from 0.204 to 0.500 mg/kg, half of samples (>0.400 mg/kg) were close to the maximum contaminant level allowed by the Chinese (0.5 mg/kg). The health risk index estimation results indicated that no particular risk of As exposure to Chinese consumers existed in L. edodes.

Effects of polysaccharides from the base of Flammulina Velutipes stipe on growth of murine RAW264.7, B16F10 and L929 cells.

A polysaccharide (FVSP) was isolated from the base of Flammuliana Velutipes stipe, and FVSP was further purified by DEAE-cellulose-52 chromatography and Sephadex G-100 size-exclusion chromatography to obtain three fractions named FVSP-1, FVSP-2 and FVSP-3. Then their activation of macrophage cell RAW 264.7 and anti-proliferative effects to the murine melanoma B16F10 and fibroblasts L929 cells were evaluated by using the cell model experiments. The results indicated that the polysaccharide fractions could increase the proliferation and phagocytic activity of macrophage significantly and play an inhibited effect on the cancer cells. Moreover, the anti-proliferative activities of FVSPs increased with the participation of the antitumor factors induced from macrophage by polysaccharides fractions. Taken together, these results indicated that three polysaccharides fractions from the base of F. Velutipes stipe may be useful as potent antitumor agents for the prevention of tumorigenesis.