Angelica gigas polysaccharide induces CR3-mediated macrophage activation and the cytotoxicity of natural killer cells against HCT-116 cells via NF-κB and MAPK signaling pathways.

Angelica gigas (A. gigas) is traditional medicinal herb that mainly exists in Korea and northeastern China. There have been relatively few studies conducted thus far on its polysaccharides and their bioactivities. We purified and described a novel water-soluble polysaccharide derived from A. gigas and investigated its immunoenhancing properties. The basic components of crude and purified polysaccharides (F1 and F2) were total sugar (41.07% - 70.55%), protein (1.12-10.33%), sulfate (2.9-5.5%), and uronic acids (0.5-31.05%) in total content. Our results demonstrated that the crude and fractions' molecular weights (Mw) varied from 42.2 to 285.2 × 103 g/mol. As the most effective polysaccharide, F2 significantly stimulated RAW264.7 cells to release nitric oxide (NO) and express several cytokines. Furthermore, F2 increased the expression of tumor necrosis factor-α (TNF-α), interferon-gamma (IFN-É£), natural killer cytotoxicity receptors (NKp44), and granzyme-B in NK-92 cells and enhanced the cytotoxicity against HCT-116 cells. In our experiments, we found that F2 stimulated RAW264.7 cells and NK-92 cells via MAPK and NF-κB pathways. The monosaccharide and methylation analysis of the high immunostimulant F2 polysaccharide findings revealed that the polysaccharide was primarily composed of 1 â†’ 4, 1 â†’ 6, 1 â†’ 3, 6, 1 â†’ 3 and 1 â†’ 3, 4, 6 galactopyranose residues, 1 â†’ 3 arabinofuranose residues, 1 â†’ 4 glucopyranose residues. These results demonstrated that the F2 polysaccharide of A. gigas which possesses potential immunostimulatory attributes, could be used to create a novel functional food.

Recurrence of ischaemic stroke using epidemiology and neuroimaging: A retrospective study in Gansu Province.

Objectives: This retrospective study aimed to investigate the clinical and imaging recurrence of ischaemic stroke (IS), and to evaluate the risk factors for recurrence. A combined clinical and imaging diagnostic model is important for stroke prevention and management. Methods and materials: In accordance with the inclusion and exclusion criteria, we retrospectively analysed consecutively hospitalised patients with acute IS at the Affiliated Hospital of Gansu University of Chinese Medicine. Based on the epidemiological and imaging results, stroke episodes were divided into four categories: clinical first episode (CFE), clinical recurrence (CR), imaging first episode (IFE), and imaging recurrence (IR). Based on the above categories and clinical practice, a joint diagnostic system for IS was established for the first time, including the following five types: IFE, IR, CFE and IFE, CFE and IR, and CR and IR. A binomial logistic regression analysis was conducted to determine the factors which contributed to CR and IR. Results: In total, 280 patients were assessed. The CR rate was 22.9% (64/280) and the IR rate was 62.9% (176/280). The only predictor of CR was hypertension (P = 0.019, odds ratio [OR] = 3.041, 95% confidence interval [95%CI] = 1.200-7.704). The factors of hypertension (P < 0.001, OR = 3.551, 95%CI = 1.781-7.080) and age (P = 0.031, OR = 1.031, 95%CI = 1.003-1.060) were predictors of IR. Conclusion: The IR rates for IS were three times higher than the CR rates. The key to preventing IR and CR in IS was the management of blood pressure. Neuroimaging examinations were important for the early detection of IFE and IR in elderly patients with hypertension. A combined clinical imaging diagnostic model was developed for the first time.

miR-126-5p Targets SP1 to Inhibit the Progression of Parkinson's Disease.

BACKGROUND: At present, symptomatic treatment may improve the life quality of Parkinson's disease (PD) patients to a certain extent but cannot completely cure PD. Therefore, it is urgent medical problem to be solved for improving the efficacy and safety of PD treatment. METHODS: SH-SY5Y and SK-N-SH cells were treated with 1-methyl-4-phenylpyridinium (MPP+) to establish PD model cells. miR-126-5p and specific protein-1 (SP1) expression levels were detected by quantitative Real-Time PCR (qRT-PCR). Western blot was applied to measure protein levels of SP1, Bax, and Bcl-2. The viabilities and apoptosis rates of treated cells were measured using cell counting kit-8 assay and flow cytometry analysis. Enzyme-linked immunosorbent assay was performed to measure TNF-α and IL-1ß releases. Interaction between miR-126-5p and SP1 was examined by dual-luciferase reporter assay. RESULTS: MPP+ treatment greatly downregulated miR-126-5p expression while upregulated SP1 expression in SH-SY5Y and SK-N-SH cells in a time- and does-dependent manner. Overexpression of miR-126-5p facilitated cell viability, while reduced cell apoptosis and inflammatory responses induced by MPP+ treatment. Moreover, SP1 was a target of miR-126-5p and could be negatively regulated by miR-126-5p. Overexpression of SP1 could reverse the effects of miR-126-5p on MPP+-administrated cells. CONCLUSION: Our results suggested that miR-126-5p attenuated the neurotoxicity induced by MPP+ in vitro through targeting SP1 (Graphical abstract), which further enhanced our understanding of the pathological mechanism of PD.

Effects of Auricularia auricula-judae polysaccharide on pasting, gelatinization, rheology, structural properties and in vitro digestibility of kidney bean starch.

Auricularia auricula-judae polysaccharide (AP) has unique molecular structures and multiple bioactivities with excellent gel-forming property and thermal tolerance. However, few researches focus on the interactions between AP and legume starches. In this study, the effects of AP on the pasting, gelatinization, rheology, microstructure, and in vitro digestibility of kidney bean starch (KBST) were evaluated. The pasting, gelling and structural properties of AP-KBST mixtures were characterized by rapid visco analyzer, rheometry, texture analyzer, laser particle analyzer, low-field nuclear magnetic resonance, Fourier transform infrared spectroscopy and scanning electron microscopy, respectively. And an in vitro method was employed to measure the digestibility of AP-KBST composites. The pasting viscosity, swelling degree of starch granules, viscoelasticity, gel strength, cold storage stability and water-retention capacity of KBST were enhanced with increasing AP concentration. The combination of AP and KBST exhibited a higher short-range ordered and a firmer and denser structure than that of KBST alone. Moreover, AP increased the contents of resistant starch and slowly digestible starch, which were positively correlated with the storage modulus and the degree of order, thereby suggesting that the formation of strong and ordered gel network structure by synergistic interactions between AP and KBST was responsible for the reduced starch digestibility.

Fractionation, structural characteristics and immunomodulatory activity of polysaccharide fractions from asparagus (Asparagus officinalis L.) skin.

The physicochemical properties, structural features and structure-immunomodulatory activity relationship of pectic polysaccharides from the white asparagus (Asparagus officinalis L.) skin were systematically studied. Using sequential ethanol precipitation, five sub-fractions namely WASP-40, WASP-50, WASP-60, WASP-70 and WASP-80 with distinct degree of esterification (DE) and molecular weight (Mw) were obtained. The Mw and DE values were decreased with the increase of the ethanol concentrations. Structurally, although 4-α-D-GalpA was the dominant sugar residue in all fractions, the molar ratios were decreased, whereas other sugar residues including arabinose- and mannose-based sugar residues overall increased with the increase of ethanol concentration. In addition, the effects of sub-fractions on the RAW 264.7 cells indicated that pectic polysaccharides with the higher DE value showed a stronger immunomodulatory activity. Moreover, the structure-activity relationship was also discussed in this study, which extends the value-added application of asparagus and its processing by-products.

Molecular insights into the inhibitory mechanisms of gallate moiety on the Aß1-40 amyloid aggregation: A molecular dynamics simulation study.

Alzheimer's disease is the most common form of neurodegenerative disease and the formation of Aß amyloid aggregates has been widely demonstrated to be the principal cause of Alzheimer's disease. Our previous study and other studies suggested that the gallate moiety played an obligatory role in the inhibition process of naturally occurring polyphenols on Aß amyloid fibrils formation. However, the detailed mechanisms were still unknown. Thus, in the present study, the gallic acid (GA) was specially selected and the molecular recognition mechanisms between GA molecules and Aß1-40 monomer were examined and analyzed by molecular dynamics simulation. The in silico experiments revealed that GA significantly prevented the conformational changes of Aß1-40 monomer with no ß-sheet structure during the whole 100 ns. By analyzing the binding sites of GA molecules to Aß1-40 monomer, we found that both hydrophilic and hydrophobic amino acid residues were participated in the binding of GA molecules to Aß1-40 monomer. Moreover, results from the binding free energy analysis further demonstrated that the strength of polar interactions was significantly stronger than that of nonpolar interactions. We believed that our results could help to elucidate the underlying mechanisms of gallate moiety on the anti-amyloidogenic effects of polyphenols at the atomic level.

Synergistic rheological behavior and morphology of yam starch and Auricularia auricula-judae polysaccharide-composite gels under processing conditions.

Polysaccharides from Auricularia auricula-judae (AP) have unique functionalities. The influence of AP-blending ratios (0.1-0.8%), temperature, pH, and ionic strength on the rheological interactions of yam starch (YS)-AP blends (YA, 6%) was investigated. YA gels showed shear-thinning behavior with flow-behavior indices of 0.28-0.37 and greater pseudoplasticity in the presence of 0.8% AP. With incremental AP addition, the viscosity and consistency indices significantly increased, whereas the activation energy decreased from 7.55 to 5.12 kJ/mol (p < 0.05). YA gels exhibited excellent thickening and heat tolerance and behaved as weak gels over an AP concentration range of 0.1-0.4% but as true gels at higher concentrations (e.g. 0.8%). The viscosity/elasticity of the combinations weakened with NaCl addition or extreme pH values. A complex 3-dimensional network structure of the YA system was confirmed by scanning electron microscopy. These results indicated that AP has great potential for functional applications in the starchy food industry.

Molecular characteristics of water-soluble extracts from Hypsizigus marmoreus and their in vitro growth inhibition of various cancer cell lines and immunomodulatory function in Raw 264.7 cells.

Water-soluble extracts isolated from Hypsizigus marmoreus and fractionated by ion-exchange chromatography were investigated to determine their molecular characteristics and biological activities. The crude and fractionated water-soluble extracts (F(1), F(2) and F(3)) consisted mostly of carbohydrates (55.8-98.9%) with varied protein contents (1.1-23.0%). Their monosaccharide levels were significantly different, including the glucose content (67.0-100%) and galactose content (0-33.0%). The backbone of each fraction was mainly glucose molecules connected with 1,4-glycosidic linkages which contained considerable amounts of glucose and/or galactose connected through 1,6-glycosidic linkages. The average molecular weight (M(w)) of the extracts varied considerably, ranging from 1,665×10(4) to 19×10(4) g/mol. Little cytotoxicity was apparent in cancer cell lines from the addition of these extracts; however, they significantly stimulated Raw 264.7 cells to release nitric oxide (NO) and prostaglandin E(2) (PGE(2)), suggesting their potential immunomodulating activities.

Effect of sulfated modification on the molecular characteristics and biological activities of polysaccharides from Hypsizigus marmoreus.

The effect of sulfated modification on polysaccharides from Hypsizigus marmoreus was examined by determining their molecular structures and bioactivities. The sulfation, which was implemented by using an orthogonal array design, produced polysaccharides with varying degrees of substitution (DS) ranging from 0.11 to 1.06. The sulfated polysaccharides exhibited a lower average molecular weight (M(w)) and considerably higher radius of gyration (R(g)) than those of native polysaccharide, suggesting that the conformation of the sulfated polysaccharides had been changed towards a more extended type. The inhibitory activity toward cancer cell growth was enhanced by treating with the sulfated polysaccharides by up to 34%, as compared to the native polysaccharide. In addition, treating with the sulfated polysaccharides increased the nitric oxide (NO) and cytokine (IL-1beta and TNF-alpha) release to levels comparable to those detected in the positive control, lipopolysaccharide (LPS), suggesting that the sulfated polysaccharides might have strong immunomodulatory activity.