Effects of slit dual-frequency ultrasound-assisted pulping on the structure, functional properties and antioxidant activity of Lycium barbarum proteins and in situ real-time monitoring process.

To improve the protein dissolution rate and the quality of fresh Lycium barbarum pulp (LBP), we optimized the slit dual-frequency ultrasound-assisted pulping process, explored the dissolution kinetics of Lycium barbarum protein (LBPr), and established a near-infrared spectroscopy in situ real-time monitoring model for LBPr dissolution through spectral information analysis and chemometric methods. The results showed that under optimal conditions (dual-frequency 28-33 kHz, 300 W, 31 min, 40 °C, interval ratio 5:2 s/s), ultrasonic treatment not only significantly increased LBPr dissolution rate (increased by 71.48 %, p < 0.05), improved other nutrient contents and color, but also reduced the protein particle size, changed the amino acid composition ratio and protein structure, and increased the surface hydrophobicity, zeta potential, and free sulfhydryl content of protein, as well as the antioxidant activity of LBPr. In addition, ultrasonication significantly improved the functional properties of the protein, including thermal stability, foaming, emulsification and oil absorption capacity. Furthermore, the real-time monitoring model of the dissolution process was able to quantitatively predict the dissolution rate of LBPr with good calibration and prediction performance (Rc = 0.9835, RMSECV = 2.174, Rp = 0.9841, RMSEP = 1.206). These findings indicated that dual-frequency ultrasound has great potential to improve the quality of LBP and may provide a theoretical basis for the establishment of an intelligent control system in the industrialized production of LBP and the functional development of LBPr.

Slit dual-frequency ultrasound-assisted pulping of Lycium barbarum fresh fruit to improve the dissolution of polysaccharides and in situ real-time monitoring.

In this study, the slit dual-frequency ultrasound-assisted pulping of fresh Lycium barbarum fruit was optimized to improve the dissolution of polysaccharides. The microscopic mechanism of polysaccharide dissolution was explored through establishing polysaccharides dissolution kinetics model and visualizing the multi-physical fields during ultrasonic process, and an in situ real-time monitoring model was established by the relationship between the chemical value and spectral information collected by near-infrared spectroscopy. The results showed that, under optimal conditions, treatment with ultrasound (28-33 kHz, 250 W, 30 min) not only significantly promoted the dissolution rate of polysaccharides in Lycium barbarum pulp (LBPPs, increased by 43.64 %, p < 0.01), reduced its molecular weight, but also improved the arabinose molar ratio, the uniformity of polysaccharide particles, and the antioxidant activity of LBPPs. Correlation analysis indicated that ultrasonic treatment is closely related to LBPPs content, particle size and scavenging capacity against superoxide anion radicals (ptotal sugar content < 0.01, pparticle size < 0.05 and psuperoxide anion scavenging < 0.05). Moreover, the in situ real-time monitoring model for the pulping process could quantitatively predict LBPPs dissolution rate and its superoxide anion radical scavenging capacity with good calibration and prediction performance (Rc = 0.9841, RMSECV = 0.0873, Rp = 0.9772, RMSEP = 0.0530; Rc = 0.9874, RMSECV = 0.1246, Rp = 0.9868, RMSEP = 0.0665). These results indicated that slit dual-frequency ultrasound has great potential in improving the quality of Lycium barbarum pulp, which may provide theoretical support for the industrial development of intelligent systems for polysaccharides preparation.

Flavonoids from Lycium barbarum Leaves Exhibit Anti-Aging Effects through the Redox-Modulation.

Lycium barbarum leaves are a kind of vegetable, and modern nutrition studies have found that they have an anti-aging function. Our study aims to investigate the anti-aging effects of Lycium barbarum leaf flavonoid (LBLF) extracts and its underlying molecular mechanism. LBLFs were purified using D101 and polyamide resin, characterized by ultraperformance liquid chromatography coupled with mass spectrometry, and administered to hydrogen peroxide (H2O2)-treated human umbilical vein endothelial cells (HUVECs) and Caenorhabditis elegans. Appropriate enrichment conditions were optimized through dynamic adsorption and desorption experiments, the content of flavonoids reached 909.84 mg/g, rutin and kaempferol being the main ones. LBLFs attenuated H2O2-induced HUVEC apoptosis, decreased reactive oxygen species and malondialdehyde production levels, increased superoxide dismutase, glutathione peroxidase and catalase activities. Furthermore, pre-treatment with LBLF increased mRNA expression of erythropoietin (EPO) and heme oxygenase-1 (HO-1) via the mitogen-activated protein kinase (MAPK) signaling pathway in HUVECs. Compared with 100 µM rutin monomer, LBLF prolonged the lifespan of Caenorhabditis elegans, enhanced their mobility in middle life stages and upregulated expression of sod-2, gcs-1 and skn-1 genes, which indicated that the anti-aging effects of LBLF were due to its redox-modulation.

Effects of Bushen-Tiaojing-Fang on the pregnancy outcomes of infertile patients with repeated controlled ovarian stimulation.

Bushen-Tiaojing-Fang (BSTJF) is commonly used to treat infertility. This study investigated the effects of BSTJF on the pregnancy outcomes of patients with repeated controlled ovarian stimulation (COS), on mitochondrial function, and on oxidative stress in ovarian granulosa cells (GCs) and follicular fluid (FF). The samples and clinical data of 97 patients, including 35 in the control group, 29 in the placebo group and 33 in the BSTJF group, were collected for this study. The mitochondrial ultrastructure, ATP content, mitochondrial DNA (mtDNA) number, 8-hydroxy-2-deoxyguanosine (8-OHdG), Mn-superoxide dismutase (Mn-SOD), glutathione peroxidase (GSH-Px) activity levels, and mRNA expression levels of Mn-SOD, GSH-Px, and nuclear factor erythroid-derived factor 2-related factor 2 (Nrf2) were analyzed. The high-grade embryo (P < 0.001), implantation (P = 0.033), and clinical pregnancy (P = 0.031) rates, as well as the ATP content (P = 0.014), mtDNA number (P = 0.035), GSH-Px activity (P = 0.004 in GCs and P = 0.008 in FF) and mRNA expression levels (P = 0.019), were significantly lower in the placebo group than in the control group, whereas the 8-OHdG content was significantly (P = 0.006 in FF) higher in the placebo group than in the control group. Compared with those in the placebo group, the high-grade embryo rate (P = 0.007), antioxidant enzyme activity (P = 0.037 and 0.036 in Mn-SOD; P = 0.047 and 0.030 in GSH-Px) and mRNA level (P < 0.001 in Nrf2, P = 0.039 in Mn-SOD and P = 0.002 in GSH-Px) were significantly higher in the BSTJF group, as were changes in mitochondrial ultrastructure, ATP (P = 0.040) and mtDNA number (P = 0.013). In conclusion, BSTJF can improve oxidative stress in patients with repeated COS and pregnancy outcomes.

Burkholderia cepacia lipase immobilized on heterofunctional magnetic nanoparticles and its application in biodiesel synthesis.

Biodiesel production using immobilized lipase as a biocatalyst is a promising process. The performance of immobilized lipase is mainly determined by supporting materials and immobilization method. To avoid the shortcomings of adsorption and covalent bonding methods, in this study, we developed a novel heterofunctional carrier of being strengthened anion exchange and weakened covalent binding to avoid activity loss and improve operational stability of the immobilized lipase. 2,3-epoxypropyltrimethylammonium chloride with epoxy and quaternary ammonium group and glutaraldehyde were grafted onto aminated magnetic nanoparticles (AMNPs) to generate a new matrix, named GEAMNP. Then Burkholderia cepacia lipase (BCL) was immobilized on GEAMNP via anion exchange and covalent bonding. The transesterification between soybean oil and methanol was used to test the activities. Activity recovery of the immobilized BCL was up to 147.4% and the corresponding transesterification activity was 1.5-fold than that of BCL powder. The immobilized lipase was further used for biodiesel production to confirm its feasibility. The fatty acid methyl esters conversion yield could reach 96.8% in the first 12 h. Furthermore, the immobilized lipase, BCL-GEAMNP showed markedly improved operational stability, better reusability and higher esters than BCL-GAMNP, where MNPs were only modified with (3-aminopropyl) triethoxysilane and glutaraldehyde.

Polysaccharides from Lycium barbarum leaves: isolation, characterization and splenocyte proliferation activity.

The polysaccharides from the fruits of Lycium barbarum have received considerable attention in previous publication, but the polysaccharides from the leaves were rarely reported. In the present work, four water-soluble polysaccharide fractions: LBP-I, LBP-II, LBP-III and LBP-IV isolated from L. barbarum leaves were purified through DEAE-Sephadex A-25. LBP-II and LBP-IV respectively showed one symmetrical peak on HPGPC with average molecular weight of 9.39 × 104 Da and 4.18 × 105 Da. UV and IR analysis of the two fractions showed the characteristics of acidic polysaccharides combined with polypeptides or proteins. GC analysis showed LBP-IV was mainly composed of rhamnose, arabinose, xylose, glucose and galactose with molar ratio of 1.61:3.82:3.44:7.54:1.00, and the uronic acid content was 47.68% (w/w) determined by sulfuric acid-carbazole method. ¹H and ¹³C NMR spectra of LBP-IV also showed the presence of carboxyl carbon and five anomeric carbons, and suggested there may be both α- and ß-anomeric configurations in this fraction. Moreover, splenocyte proliferation activity assay showed that LBP-IV significantly enhanced the proliferation of splenocyte stimulated by ConA or LPS, indicating the fraction has the beneficial effect on immunostimulating activity.

[Polysaccharides from Scurrula parasitica L. inhibit sarcoma S180 growth in mice].

To study the anti-tumor activity of Scurrula parasitica polysaccharides (SP). Water extraction and ethanol precipitation were used to isolate SP from S. parasitica leaf. S180, K562 and HL-60 cell lines proliferation inhibition by SP were detected by MTT assay. The expressions of Ki-67, Cyclin D1, Bax and Bcl-2 protein in the sarcoma S180 tissues were detected by immunohistochemistry technique to approach the anti-tumor mechanism of SP+ SP could not inhibit cancer cell proliferation. SP ip could inhibit the growth of sarcoma S180 in mice, 100 mg x kg(-1) x d(-1). SP ip was the optimal dose on inhibiting S180 growth, with the tumor inhibition rate of 54%. The expression of Ki-67, Cyclin D1, Bax and Bcl-2 protein in the sarcoma S180 tissues were detected by immunohistochemistry technique to approach the anti-tumor mechanism of SP. The result showed that SP could down-regulate the expression of Ki-67, CyclinD1 and Bcl-2 protein, and up-regulate the expression of Bax protein. It indicted that inhibiting cancer cell proliferation and promoting cancer cell apoptosis in vivo maybe one of the anti-cancer mechanisms of SP.

[Study on cytotoxic activities on human leukemia cell line HL-60 by flavonoids extracts of Scurrula parasitica from four different host trees].

OBJECTIVE: To compare the anticancer effects of flavonoids extracts of Scurrula parasitica from different host trees in vitro. METHOD: 80% ethanol extracts of S. parasitica parasitizing on Nernium indicum, Morus alba, Opsmanthus fragrans, and Sapindus mulorossi were purified by polyamides column chromatography, and the eluates of 30%, 50%, 70% and 90% ethanol were mixed as flavonoids extracts. Human acute myeloid leukemia cell line HL-60 was used to evaluate the cytotoxicity induced by flavonoids extracts of S. parasitica L with MTT assay. Apoptosis was detected by AO/EB fluorescence staining and DNA fragmentation analysis, apoptosis rates and cell cycle distribution were detected by flow cytometry analysis. RESULT: Extract of S. parasitica parasitizing on N. indicum (NISPEX) was the most sensitive to HL-60 cells of the 4 different host trees, the IC50 value being 0.60 mg x L(-1); and extract of S. parasitica parasitizing on M. alba took the second place, the IC50 value, being 2.49 mg x L(-1); extract of S. parasitica parasitizing on O. fragrans had no effectiveness as high as 50 mg x L(-1) concentration. NISPEX induced HL-60 cell apoptosis and inhibited the cell proliferation in dose and time-dependent manner. Cell cycles were arrested at G0-G1 phase after treated with NISPEX. CONCLUSION: Anticancer effects of S. parasitica correlated with the host trees. Flavonoids extracts of S. parasitica parasitizing on N. indicum exhibited comparatively better anticancer activity in vitro among the host trees studied. NISPEX is found to be a good candidate for anticancer.