The polysaccharide extract (PE) of Uyghur medicinal preparation Alhagi-honey was prepared by water extraction and alcohol precipitation method. The purified polysaccharide AP1-1 was obtained from PE by macroporous adsorption resin chromatography, DEAE cellulose chromatography, and Sephadex gel chromatography; the homogeneity and the molecular weight of AP1-1 were determined by gel filtration; and the acid hydrolysis, periodate oxidation, Smith degradation, and NMR analysis were used to analyze the chemical structure of AP1-1. The result showed that AP1-1 was a homogeneous polysaccharide, whose relative molecular weight was 9.97 × 10(4). Through high-performance capillary electrophoresis analysis, we found that its molecular structure was composed of mannose, glucose, galactose, and galacturonic acid with a molar ratio of about 1.1:1.9:3.9:2.1. The main chain of AP1-1 was mainly made up of → 4)β-d-GalpA-(1 → 4)β-d-GalpA-(1 → 4)-β-d-Galp-(1 → 4)-β-d-Galp-(1 → 6)α-d-Glcp-(1 → 4)α-d-Glcp(1 → , while the side chain is composed of → 6)-α-d-Glcp and 2-CH3-α-d-Man.
Abstract. The in vitro immune activities of Saccharum Alhagi polysaccharides (SAP) have been previously studied. The present study aimed to investigate the effects of SAP-1 and SAP-2 on the activity of RAW264.7 mouse macrophages. RAW264.7 cells were treated with 150, 300 and 600 mg/l concentrations of SAP-1 (a 50% alcohol precipitation) and SAP-2 (an 80% alcohol precipitation) or with 10 mg/l lipopolysaccharide. Untreated cells were used as a negative control. An MTT assay was used to detect the proliferation of the cells, and Hoechst 33528 staining was conducted in order to visualize the cell nuclei. Additionally, the Griess method was used to measure nitric oxide (NO) levels. A neutral red uptake assay was performed to determine the phagocytic activity of the macrophages, and ELISAs were performed to detect cytokine levels. Furthermore, reverse transcription-quantitative polymerase chain reaction was used to measure the mRNA expression of certain cytokines. The results demonstrated that SAP increased the proliferative activity and activated the immune function of RAW264.7 cells, and was lacking in cytotoxicity. In addition, SAP-1 exhibited a stronger effect in promoting RAW264.7 cell proliferation than did SAP-2. Furthermore, SAP-1 and SAP-2 significantly increased the level of NO, with the effect of SAP-1 being stronger than that of SAP-2. SAP-1 increased the phagocytic activity of RAW264.7 cells and promoted the secretion of the cytokines interleukin (IL)-1β, IL-2 and tumor necrosis factor (TNF)-α by RAW264.7 cells, with an effect that was stronger than that of SAP-2. Finally, different concentrations of SAP-1 or SAP-2 had distinct effects in upregulating the expression of TNF-α, IL-1β, nuclear factor-κB and inducible nitric oxide synthase mRNA. The results of the present study demonstrate that SAP is capable of enhancing the immune activity of mouse macrophages.
A novel fluorescence imaging platform based on a high-throughput immunosensor chip and a DNA dendrimer capped with plenty of fluorescent dyes was proposed for ultrasensitive quantitation of cardiac troponin T.
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