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A neutral polysaccharide designated as CMDP-1a (molecular mass 9.263 kDa) was isolated from Cucurbita moschata Duch through hot water extraction, ethanol precipitation, and column chromatography. On the basis of methylation, fourier-transform infrared, monosaccharide composition, and one- and two-dimensional nuclear magnetic resonance spectroscopy analyses, the structure of CMDP-1a was determined to be a backbone composed of α-1,4 linked glucopyranosyl residues with α-Glcp residue linkage at backbone C-6. Atomic force microscopy and scanning electron microscopy analyses revealed that CMDP-1a had a spherical conformation in solution. In immunostimulation assays, CMDP-1a promoted the proliferation of RAW 264.7 macrophages and significantly enhanced their pinocytic and phagocytic capacity. Furthermore, CMDP-1a induced the M1 polarization of original macrophages and the conversion of macrophages from M2 to M1, thereby modulating the balance of M1/M2 macrophages. These results indicated that CMDP-1a might be a potential immunomodulator for food purposes.

Pumpkin polysaccharides with various bioactivities are mainly taken orally, thus detailed knowledge of the intestinal transport of which are essential for understanding its bioactivities. The Caco-2 cells monolayer model (mimic intestinal epithelium) was successfully constructed and Cucurbia moschata polysaccharides (PPc-F) were successfully conjugated with fluorescein isothiocyanate (FITC) to evaluate the transcellular transport mechanism and subcellular localization of PPc. The transport process of PPc-F was energy-dependent, and a moderately-absorbed biological macromolecule according to the apparent permeability coefficients (Papp) value. The endocytosis process of PPc-F in Caco-2 cells included the clathrin- and caveolae (or lipid draft)-medicated routes. And the translocation process was related to endoplasmic reticulum (ER), golgi apparatus (GA), tubulin and the acidification of endosomes. As for the intracellular location of PPc-F, it was mainly accumulated in ER. The study provided an understanding of the transmembrane transport of PPc-F, and could help studying the mechanisms of its effects.

To date, a safe and reliable treatment of osteoarthritis (OA) has not yet been announced. Inflammatory response and degradation of the articular extracellular matrix (ECM) induced by IL-1β are important pathological characteristics of OA. Laquinimod is a quinoline-3-carboxamide and a novel oral immunomodulatory compound in clinical use. However, whether laquinimod has a beneficial effect in OA is not known. In our research, we found that laquinimod could ameliorate IL-1β-induced generation of ROS and improve mitochondrial function by increasing mitochondrial membrane potential (ΔΨm). Furthermore, treatment with laquinimod suppressed IL-1β-induced production of TNF-α and IL-6. Notably, laquinimod prevented the degradation of type II collagen by inhibiting MMP-3 and MMP-13. Meanwhile, the presence of laquinimod attenuated the reduction in aggrecan by mediating ADAMTS-4 and ADAMTS-5. Mechanistically, laquinimod ameliorated IL-1β-induced inflammation and degeneration of ECM by suppressing the activation of NF-κB. Taken together, our findings reveal that laquinimod possesses a beneficial effect against IL-1β insults in human chondrocytes, implying an important role of laquinimod in OA.

This study investigated an efficient and recyclable approach for purification of crude pumpkin oligosaccharide (POS) by macroporous resins. Five resins with different physical and chemical properties were tested for decoloration of POS. In virtue of its higher decoloration ratio (92.6%) and POS recovery ratio (81.3%), the macroporous resin DM28 was considered to a better selection. Depending on the changes of molecular weight, part of the monosaccharides in crude POS were removed simultaneously after decoloration by DM28. Operating conditions were also determined by the dynamic breakthrough and desorption curves. Moreover, UV/vis spectroscopy and Fourier transform infrared results revealed that most of the colored impurities and proteins can be removed, but the characteristic groups of the POS exhibited no significant difference. Compared with traditional methods, DM28 resin is superior in decoloration efficiency, pigment recovery and oligosaccharide recovery. This research contributes to further exploration on the structure and function of POS.

A pectic polysaccharide (named CMDP-4b) with a molecular weight of 31.97 kDa was extracted from Cucurbita moschata Duch and purified by column chromatography. On the basis of methylation, Fourier-transform infrared, monosaccharide composition, and one- and two-dimensional nuclear magnetic resonance spectroscopy analyses, the structure of CMDP-4b was determined to be composed of an α-1,4-linked homogalacturonan backbone, which was slightly acetylated and highly methyl-esterified, and branched at the O-3 position of the →4)-α-D-GalpA-6-OMe-(1→. Immunomodulatory assays showed that CMDP-4b not only induced the secretion of nitrous oxide and cytokines (i.e. IL-1β, TNF-α, and IL-6) but also promoted pinocytic and phagocytic activities of macrophages, suggesting that CMDP-4b possessed immunomodulatory activity. Moreover, toll-like receptor 4 and complement receptor 3 may play a critical role in CMDP-4b-induced macrophage activation through the NF-κB and the MAPKs signaling pathways. Our study provides the molecular basis for the potential use of CMDP-4b as a natural immunostimulant.

In this paper, subcritical water (SCW) was applied to modify pumpkin (Cucurbita moschata Duchesne ex Poiret) polysaccharides, and the properties and antioxidant activity of pumpkin polysaccharides were investigated. SCW treatments at varying temperature led to changes in the rheological and emulsifying properties of pumpkin polysaccharides. SCW treatments efficiently degraded pumpkin polysaccharides and changed the molecular weight distribution. Decreases in intrinsic viscosity, viscosity-average molecular weight, and apparent viscosity were also observed, while the activation energy and flow behavior indices increased. The temperature of SCW treatment has a great influence on the linear viscoelastic properties and antioxidant activity of pumpkin polysaccharides. Pumpkin polysaccharides solution treated by SCW at 150 &deg, C exhibited the highest emulsifying activity and antioxidant activity, which was probably due to a broader molecular mass distribution and more reducing ends exposed after treatment. Scanning electron microscopy showed that SCW treatment changed the microstructure of pumpkin polysaccharides, resulting in the exposure of bigger surface area. Our results suggest that SCW treatment is an effective approach to modify pumpkin polysaccharides to achieve improved solution properties and antioxidant activity.

The novel natural low-molecular-mass polysaccharide (SLWPP-3) from pumpkin (Cucurbia moschata) was separated from the waste supernatant after macromolecular polysaccharide production and purified using a DEAE cellulose-52 column and gel-filtration chromatography. Chemical and instrumental studies revealed that SLWPP-3 with a molecular mass of 3.5 kDa was composed of rhamnose, glucose, arabinose, galactose and uronic acid with a weight ratio of 1: 1: 4: 6: 15, and primarily contained →3,6)-β-d-Galp-(1→, →4)-α-GalpA-(1→(OMe), →4)-α-GalpA-(1→, →2,4)-α-d-Rhap-(1→, →3)-β-d-Galp-(1→, →4)-α-d-Glcp, and →4)-β-d-Galp residues in the backbone. The branch chain passes were connected to the main chain through the O-4 atom of glucose and O-3 atom of arabinose. Physiologically, the ability of SLWPP-3 to inhibit carbohydrate-digesting enzymes and DPPH and ABTS radicals, as well as protect pancreatic β cells from oxidative damage by decreasing MDA levels and increasing SOD activities, was confirmed. The findings elucidated the structural types of pumpkin polysaccharides and revealed a potential adjuvant natural product with hypoglycemic effects.

Pumpkin polysaccharide is able to alleviate diabetes, but understanding of the underlining mechanism is still limited. In this study, we hypothesized that the alleviating effects of pumpkin polysaccharide is modulated via changes in the gut microbiota and short-chain fatty acid (SCFA) production in type 2 diabetic rats. After the type 2 diabetic model successfully was established, three groups of high-fat diet induced diabetic rats were intragastrically administered pumpkin polysaccharide, metformin, or saline solution respectively. We utilized 16S rRNA gene sequencing and multivariate statistics to analyze the structural and key species of gut microbiota in the type 2 diabetic rats. The results revealed that pumpkin polysaccharide alleviated the type 2 diabetes by improving the insulin tolerance and decreasing the levels of serum glucose (GLU), total cholesterol (TC), and low-density lipoprotein (LDL-C), while increasing the levels of high-density lipoprotein (HDL-C). Simultaneously, pumpkin polysaccharide changed the structure of gut microbiota and had selective enrichment in key species of Bacteroidetes, Prevotella, Deltaproteobacteria, Oscillospira, Veillonellaceae, Phascolarctobacterium, Sutterella, and Bilophila. The correlations between the key species and SCFA production indicated the underlining mechanisms of pumpkin polysaccharide on type 2 diabetes.

Zhaotao Wang,1,* Guoyong Yu,2,* Zhi Liu,3 Jianwei Zhu,4 Chen Chen,1 Ru-en Liu,3 Ruxiang Xu1 1Department of Neurosurgery, Affiliated Bayi Brain Hospital, General Army Hospital, Southern Medical University, Beijing, China; 2Department of Neurosurgery, Jiangxi Provincial People’s Hospital, Nanchang, China; 3Department of Neurosurgery, Peking University People’s Hospital, Peking University, Beijing, China; 4Department of Neurosurgery, Affiliated Hospital of Traditional Chinese Medicine, Southwest Medical University, Luzhou, China *These authors contributed equally to this work Background: Paeoniflorin, a polyphenolic compound derived from Radix Paeoniae Alba (Paeonia lactiflora), has exhibited anticancer activity in various human cancers, including glioblastoma. However, the mechanisms underlying the effects of this compound have not been fully elucidated. Toll-like receptor 4 (TLR4) plays an important role in the regulation of cancer cell proliferation and progression, and high TLR4 expression in glioblastoma specimens is associated with a poor prognosis. The present study aimed to investigate whether paeoniflorin suppresses glioblastoma via inhibition of TLR4 expression.Methods: CCK-8 experiments and clone formation assay were performed to detect the cell proliferation. Western blotting was used to analyze protein expression levels. Detection of Triad3A binding with TLR4 was assessed by the immunoprecipitation. Orthotopic xenograft mouse model was used to evaluate the effect of paeoniflorin in vivo. MST was used to analyze the interaction between paeoniflorin and TLR4 protein.Results: In our study, we found that paeoniflorin effectively inhibited glioblastoma growth and suppressed TLR4 protein levels, as well its downstream effectors both in vivo and in vitro. Moreover, when overexpressed TLR4 in glioblastoma abolished the effects of paeoniflorin on cell proliferation, migration, and invasion. Furthermore, we found that paeoniflorin decreased TLR4 protein through ubiquitination proteasome pathway (UPP)-mediated degradation in glioblastoma cells. Mechanistically, paeoniflorin promoted Triad3A to conjugate with TLR4, resulting in degradation. In addition, Triad3A-shRNA abolished paeoniflorin-enhanced UPP-mediated TLR4 degradation. Finally, we found that paeoniflorin could directly bind with TLR4 protein as assessed by MST assay.Conclusion: Our study is the first to identify a novel mechanism for the antitumor activity of paeoniflorin, specifically: it decreases tumor growth by directly targeting TLR4 and modulating the TLR4/Triad3A-dependent axis, leading to TLR4 protein degradation and inhibition of glioblastoma cell progression in vitro and in vivo. Our current findings indicate that paeoniflorin is a potential glioblastoma therapeutic agent due to its Triad3A-dependent ubiquitin degradation of TLR4. Keywords: paeoniflorin, glioblastoma, TLR4, ubiquitin, Triad3A&nbsp

Bioactive compounds present in foods could have beneficial effects on human health. In this study, we report the capacity of peptides released from oat, highland barley, and buckwheat proteins after enzymatic digestion to inhibit platelet aggregation in vitro. All hydrolysates showed high antiplatelet activity, with IC50 values of 0.282mg/ml (oat flour gastrointestinal hydrolysate, 6h) to 2.496mg/ml (highland barley glutelin tryptic hydrolysate, 14h) in a dose-dependent manner. Thirty-eight peptides with more than seven residues were identified in the tryptic hydrolysates of oat globulin. Results of computational modeling revealed that nine peptides, including ALPIDVLANAYR, EFLLAGNNKR, GEEFGAFTPK, QLAQIPR, LQAFEPLR, ALPVDVLANAYR, GEEFDAFTPK, QKEFLLAGNNK, and TNPNSMVSHIAGK bound the cyclooxygenase-1 active centers with low binding energy (-6.5 to -7.5kcal/mol). This is the first report to identify antiplatelet peptides from grain hydrolysates and the binding modes at the molecular level, leading to their possible use as functional food ingredients to prevent thrombosis.

As a new platform of systems biology, metabolomics provides a powerful approach to discover therapeutic biomarkers and mechanism of metabolic disease. Type 2 diabetes mellitus (T2DM) is a global metabolic disease, thus, a urinary metabolomics profiling was analyzed to study the anti-diabetic effects and mechanism of stachyose (ST) on high-fat diet- and low dose streptozotocinc-induced T2DM rats. The results showed that ST treatment regulated the level of insulin, low-density lipoprotein cholesterol, and triglycerides, which demonstrates improvement in T2DM on ST treatment. Urinary samples from the ST and T2DM group were enrolled in metabolomics study, 21 differential metabolites were identified from urinary metabolomics analysis, indicating that the ST treatment partly exerted the anti-diabetes activity by regulating energy metabolism, gut microbiota changes and inflammation. A metabolomics strategy is both suitable and reliable for exploring the anti-diabetes effects and understanding the mechanisms of ST treatment against T2DM.

Paeoniflorin (PF), as one of the important valid natural compounds of the total glucosides of peony, has displayed a potential effect in cancer prevention and treatment. Aggressive migration and invasion, as an important process, can contribute to tumor progression through infiltrating the surround normal tissue. Actin cytoskeleton rearrangement plays a key role in cells migration and invasion, involving multiple signal pathways. HGF/c-Met signal, as an important couple of oncoprotein, has been demonstrated to regulate actin cytoskeleton rearrangement. In our study, we aim to explore whether paeoniflorin can inhibit migration and invasion and actin cytoskeleton rearrangement via regulation of HGF/c-Met/RhoA/ROCK signal. Various approaches were applied to demonstrate the mechanism of paeoniflorin-mediated anticancer effect, including cell wound healing assay, invasion assay, immunofluorescence staining and transfection, and western blotting. We observed that paeoniflorin inhibited HGF-induced migration and invasion and actin cytoskeleton rearrangement in glioblastoma cells. Furthermore, the inhibition of HGF-induced migration and invasion and actin cytoskeleton rearrangement involved c-Met-mediated RhoA/ROCK signaling in glioblastoma. Thus, our study proved that paeoniflorin could inhibit migration and invasion and actin cytoskeleton rearrangement through inhibition of HGF/c-Met/RhoA/ROCK signaling in glioblastoma, suggesting that paeoniflorin might be a candidate compound to treat glioblastoma.

Insoluble dietary fiber (DF) extracted from carrot pomace was modified by complex enzyme method, ultrafine comminution and high hydrostatic pressure and their structural, physicochemical, and functional properties were evaluated. Results showed that complex enzyme method increased the content of soluble DF to 15.07%, and the cholesterol adsorption capacity peaked; ultrafine comminution greatly improved the total antioxidant activities, DPPH radical scavenging capacity and antioxidant abilities in linoleic acid system of DF; high hydrostatic pressure led to a significant increase in capacities of water retention (7.14 g/g, 600 MPa), water swelling (10.02 mL/g, 500 MPa), oil retention (2.35 g/g, 500 MPa), cation exchange (2.29 mmol/g, 600 MPa), and glucose adsorption (2.634 mmol/g, 400 MPa). Furthermore, SEM showed that surface structures of all modified DF were loose. Overall, three kinds of modification methods have shown their respective advantages. Modified DF from carrot pomace can be applied as a functional ingredient in diverse food products.

The fungicidal mechanism of chlorine dioxide on Saccharomyces cerevisiae was investigated. During S. cerevisiae inactivation by ClO2, protein, DNA, and ion leakage, enzyme activity, genomic DNA structure, and cell ultrastructure were examined. Protein and DNA leakages were not observed, while ion leakages of K+, Ca2+, and Mg2+ were detected and were related to the inactivation rate. The glucose-6-phosphate dehydrogenase, citrate synthase, and phosphofructokinase activities were inhibited and were also correlated with the inactivation rate. Genomic DNA structure was not damaged except for an extremely high ClO2 concentration (100 mg L-1). Electron micrographs showed that cell surface damage was pronounced and disruption in inner cell components was also apparent. The ion leakage, the inhibition of key enzyme activities of metabolic pathway, and the alteration of cell structure were critical events in S. cerevisiae inactivation by ClO2.

Vinegar has been used as both a common seasoning and a traditional Chinese medicine. Sorghum vinegar is an excellent source of physiological substances with multiple health benefits.To evaluate the antiplatelet aggregation activity of alditols and monosaccharides extracted from sorghum vinegar and analysis its mechanism.Alditol and monosaccharide extract (AME) from sorghum vinegar was first evaluated for antiplatelet activity using the turbidimetric method. Blood was collected from healthy volunteer donors. The platelet aggregation was induced by arachidonic acid (AA), collagen, adenosine diphosphate (ADP) and thrombin in vitro. AME was divided into three experimental groups with the concentration were 0.10, 0.25 and 0.50 mg/mL. In order to determine the inhibitory activity of AME on COX1, TXS and TXA2 production experiments were conducted using the COX1, TXS and TXB2 EIA kit. Computational docking was used to find the docking pose of monosaccharides and alditols with COX1.AME showed significant induction of antiplatelet activity by arachidonic acid (AA), collagen, adenosine diphosphate (ADP) and thrombin in a concentration-dependent manner (p0.05). AME (0.50 mg/mL) reduced the AA-induced aggregation rate to 10.35%±0.46%, which was comparable to acetylsalicylic acid (aspirin, ASA) (0.50 mg/mL, 6.35%±0.58%), a medical standard. Furthermore, AME strongly inhibited cyclooxygenase-1 (COX1) and thromboxane-A2 synthase (TXS), and subsequently attenuated thromboxane-A2 (TXA2) production. These findings indicated that AME attenuates platelet aggregation through the AA metabolism pathway. Computational docking showed that alditols (L-erythritol, L-arabitol, xylitol and D-sorbitol), monosaccharides (D-glucopyranose, D-fructofuranonse, D-xylopyranose, D-galactopyranose and D-ribose), ethyl glucoside and 3,4-(methylenedioxy) mandelic acid could dock directly into the active site of COX1.Alditols and monosaccharides from sorghum vinegar inhibit multiple steps in the platelet aggregation pathway, and may be beneficial for the treatment of cardiovascular diseases.

Hawthorn seed (HS), an important by-product of the Hawthorn industry, is rich in potentially health-promoting flavonoids compounds. In this paper, the ultrasound-assisted extraction (UAE) of FC from HS was investigated. Important variables and their levels were obtained using Plackett-Burman (PB) design and Box-Behnken (BB) design. A mathematical model was developed to show the effects of each variable and their combinatorial interactions on extraction yield of FC. A high coefficient of determination (R(2) = 91.26%) indicated good agreement between the experimental and predicted values of FC yield. The optimum levels of these significant parameters were determined using response surface methodology (RSM), which revealed these as follows: ultrasound temperature 65 °C, ultrasonic time 37 min, extraction temperature 91 °C, extraction time 1.5h, solid-liquid ratio of 1:18, and 72% ethanol. Under the optimum condition, the UAE rate of FC was up to 91.7%, and the yield of FC was 16.45 ± 0.02 mg/g (P