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In the present study, we studied anti-Alzheimer′s disease (AD) activities of chondroitin sulfate (CS) oligosaccharides with different molecular weights. CS from shark cartilage was degraded by a recombinant CS endolyase, chondroitinase ABC I (CHSase ABC I), and CS disaccharide (DP2), tetrasaccharide (DP4), hexasaccharide (DP6), octasaccharide (DP8), decasaccharide (DP10) and dodecasaccharide (DP12) were obtained by separation with gel filtration. Anti-AD activities of CS oligosaccharides were assessed using Aβ-injured SH-SY5Y cells and BV2 cells. It was shown that CS oligosaccharides could block Aβ-induced oxidative stress, mitochondrial dysfunction and activation of intrinsic apoptotic pathway for SH-SY5Y cells. Furthermore, these activities increased with the increase of molecular weights. For Aβ-injured BV2 cells, CS oligosaccharides inhibited oxidative stress, the production of proinflammatory cytokines and the activation of toll-like receptor pathway, and CS DP2 had the best activity among them. In conclusion, CS oligosaccharides suppressed Aβ-induced oxidative stress and relevant injury in vitro, and these effects had different relationships with the molecular weights of CS oligosaccharides for different cell lines, which might be caused by different mechanisms. [ABSTRACT FROM AUTHOR]

Hyaluronic acid (HA), a natural high molecular weight polysaccharide, is produced by Streptococcus zooepidemicus. However, Streptococcus has several drawbacks including its potential to produce exotoxins, so there is demand for an alternative HA source. Here, a recombinant HA biosynthesis operon, as well as the HA biosynthesis operon of S. zooepidemicus were introduced into L. lactis using the nisin-controlled expression system, respectively. HA was successfully synthesized by recombinant L. lactis. Furthermore, overexpression of the endogenous enzymes directing the synthesis of precursor sugars was effective at increasing HA production, and increasing the supply of UDP-activated monosaccharide donors aided synthesis of monodisperse HA polysaccharides. Besides GRAS host strain ( L. lactis) and NICE system, the selecting marker ( lacF gene) of the recombinant strain is also food grade. Therefore, HA produced by recombinant L. lactis overcomes the problems associated with Streptococcus and provides a source of food-grading HA appropriate for widespread biotechnological applications. [ABSTRACT FROM AUTHOR]

Currently, there is no analytical method for the quantification of hemocoagulase agkistrodon (HCA) in pharmaceutical preparations. This study presents a pre-column derivatization method for the quantification of HCA, a compound extracted from the venom of Agkistrodon acutus, in a pharmaceutical preparation (trade name Suling). In the proposed method, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate was used to tag the HCA substrate, and the derivatives were analyzed by high-performance liquid chromatography with fluorescence detection. Complete and homogeneous derivatization of HCA was confirmed by matrix-assisted laser desorption/ionization–time-of-flight mass spectrometry analysis. The specificity of the method was validated by forced degradation, and interference was assessed using a placebo. Under the optimum chromatographic conditions, the calibration curve was linear over a range of 10 to 500 ng/mL, featuring a correlation coefficient of 0.9999. The limits of detection and quantification of the method were 0.57 and 1.6 ng/mL, respectively. The percentage recovery of HCA in quality control samples ranged from 97.49 to 99.15%. Overall, this novel method can be applied to the quantitative determination of HCA in pharmaceutical preparations. [ABSTRACT FROM AUTHOR]

A simple and accurate capillary electrophoresis (CE) method was developed to simultaneously separate and quantify heparin, chondroitin sulfate and hyaluronic acid. The relative standard deviations (intra-day) of migration time, peak height and peak area for heparin, chondroitin sulfate and hyaluronic acid were lower than 1.11, 5.45 and 2.82%, respectively. The limits of detection of heparin, chondroitin sulfate and hyaluronic acid were 0.91, 0.12 and 9.04 × 10−3 mg/mL, respectively. The developed electrophoretic method was successfully applied to the analysis of commercial drug products and biological samples containing chondroitin sulfate and/or hyaluronic acid. The recoveries for chondroitin sulfate and hyaluronic acid were in the range of 95.9 ∼107.0%. This was the first time the content of hyaluronic acid in the synovial fluids from osteoarthritic rabbits was investigated by CE. The results suggested that hyaluronic acid in the synovial fluids from osteoarthritic rabbits may be further metabolized and the administration of chondroitin sulfate or hyaluronic acid could affect the content and metabolism of hyaluronic acid in the synovial fluids. The developed CE method was simple to implement without sample pretreatment such as depolymerisation, very repeatable and easily transferred from lab to lab. [ABSTRACT FROM PUBLISHER]