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, Volume 17 Issue 6 Previous Issue    Next Issue
Articles
Magnolol pretreatment attenuates heat stress-induced IEC-6 cell injury
Chen Mei, Sha-sha He, Peng Yin, Lei Xu, Ya-ran Shi, Xiao-hong Yu, An Lyu, Feng-hua Liu, Lin-shu Jiang
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 413-424.   https://doi.org/10.1631/jzus.B1500261
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Objective: Heat stress (HS) is an important environmental stressor that adversely influences livestock during the summer. The aim of this study was to investigate whether magnolol protects against HS-induced intestinal epithelial cell injury. Materials and methods: An intestinal epithelial cell line (IEC-6) was subjected to HS at 42 °C, with and without magnolol pretreatment. Cell injury was detected by monitoring lactate dehydrogenase (LDH) release. MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay was used to assess cell proliferation and viability, including identifying effective concentrations of magnolol. Flow cytometry confirmed G1-phase cell-cycle arrest and its alleviation by magnolol. Active DNA synthesis was measured by incorporation of nucleic acid 5-ethynyl-2\'-deoxyuridine (EdU). G1-phase cell-cycle-related gene expression was assessed by real-time reverse transcription polymerase chain reaction (RT-PCR) and levels of G1-phase-related proteins by Western blotting. Results: HS induced IEC-6 cell injury and decreased cell viability, as demonstrated by data from LDH and MTS assays, respectively. Based on a number of criteria, IEC-6 cells subjected to HS were arrested in the G1 phase of the cell cycle. Magnolol pretreatment decreased HS-induced cell injury through relief of this cell-cycle arrest. Conclusions: Magnolol pretreatment attenuates HS-induced injury in IEC-6 cells. Magnolol is potentially promising as a protective strategy for HS in livestock.
Inhibitory effect of Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts on lipopolysaccharide-stimulated inflammatory response in RAW 264.7 cells
Wei Xu, Qing Zhou, Yong Yao, Xing Li, Jiu-liang Zhang, Guan-hua Su, Ai-ping Deng
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 425-436.   https://doi.org/10.1631/jzus.B1500213
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Blueberries are a rich source of anthocyanins, which are associated with health benefits contributing to a reduced risk for many diseases. The present study identified the functional Gardenblue blueberry (Vaccinium ashei Reade) anthocyanin extracts (GBBAEs) and evaluated their capacity and underlying mechanisms in protecting murine RAW 264.7 cells from lipopolysaccharide (LPS)-stimulated inflammation in vitro. Enzyme-linked immunosorbent assay (ELISA) kit results showed that GBBAEs significantly inhibited the production of nitric oxide (NO), prostaglandin E2 (PGE2), interleukin-6 (IL-6), IL-1β, and interferon-γ (INF-γ). Real-time polymerase chain reaction (PCR) analysis indicated that the mRNA expression levels of IL-6, IL-1β, tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and cyclooxygenase 2 (COX-2) were suppressed in LPS-stimulated RAW 264.7 cells. Additionally, Western blot analysis was used to evaluate the relative protein expression levels of COX-2 and nuclear factor-κB p65 (NF-κBp65). All these results suggested the potential use of GBBAEs as a functional food for the treatment of inflammatory diseases.
Myricetin protects against diet-induced obesity and ameliorates oxidative stress in C57BL/6 mice
Hong-ming Su, Li-na Feng, Xiao-dong Zheng, Wei Chen
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 437-446.   https://doi.org/10.1631/jzus.B1600074
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Background: Myricetin is a naturally occurring antioxidant commonly found in various plants. However, little information is available with respect to its direct anti-obesity effects. Objective: This study was undertaken to investigate the effect of myricetin on high-fat diet (HFD)-induced obesity in C57BL/6 mice. Results: Administration of myricetin dramatically reduced the body weight of diet-induced obese mice compared with solely HFD-induced mice. Several parameters related to obesity including serum glucose, triglyceride, and cholesterol were significantly decreased in myricetin-treated mice. Moreover, obesity-associated oxidative stress (glutathione peroxidase (GPX) activity, total antioxidant capacity (T-AOC), and malondialdehyde (MDA)) and inflammation (tumor necrosis factor-α (TNF-α)) were ameliorated in myricetin-treated mice. Further investigation revealed that the protective effect of myricetin against HFD-induced obesity in mice appeared to be partially mediated through the down-regulation of mRNA expression of adipogenic transcription factors peroxisome proliferator-activated receptor γ (PPARγ) and CCAAT/enhancer-binding protein α (C/EBPα), and lipogenic transcription factor sterol regulatory element-binding protein 1c (SREBP-1c). Conclusions: Consumption of myricetin may help to prevent obesity and obesity-related metabolic complications.
Apigenin-7-O-β-D-glycoside isolation from the highly copper-tolerant plant Elsholtzia splendens
Hong-yun Peng, Xue-hong Zhang, Jin-zhong Xu
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 447-454.   https://doi.org/10.1631/jzus.B1500242
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Elsholtzia splendens (Lamiaceae) is a copper-tolerant plant species growing on copper deposits in the south of China. Chromatographic separation of n-BuOH extracts from the flowering aerial biomass afforded apigenin-7-O-β-D-glycoside, using macroporous resin, Sephadex™ LH-20 gel, polyamide resin as well as preparative high-performance liquid chromatography (P-HPLC) columns. Chemical structure was elucidated using HPLC/ESI-MS (electrospray ionization-mass spectrometry), Fourier transform infrared (FTIR), and 1D- and 2D-nuclear magnetic resonance (NMR). Apigenin-7-O-β-D-glycoside could be the post-harvesting product from E. splendens biomass.
Characterization of β-glucosidase from Aspergillus terreus and its application in the hydrolysis of soybean isoflavones
Feng-ying Yan, Wei Xia, Xiao-xu Zhang, Sha Chen, Xin-zheng Nie, Li-chun Qian
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 455-464.   https://doi.org/10.1631/jzus.B1500317
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An extracellular β-glucosidase produced by Aspergillus terreus was identified, purified, characterized and was tested for the hydrolysis of soybean isoflavone. Matrix-assisted laser desorption/ionization with tandem time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS) revealed the protein to be a member of the glycosyl hydrolase family 3 with an apparent molecular mass of about 120 kDa. The purified β-glucosidase showed optimal activity at pH 5.0 and 65 °C and was very stable at 50 °C. Moreover, the enzyme exhibited good stability over pH 3.0–8.0 and possessed high tolerance towards pepsin and trypsin. The kinetic parameters Km (apparent Michaelis-Menten constant) and Vmax (maximal reaction velocity) for p-nitrophenyl-β-D-glucopyranoside (pNPG) were 1.73 mmol/L and 42.37 U/mg, respectively. The Km and Vmax for cellobiose were 4.11 mmol/L and 5.7 U/mg, respectively. The enzyme efficiently converted isoflavone glycosides to aglycones, with a hydrolysis rate of 95.8% for daidzin, 86.7% for genistin, and 72.1% for glycitin. Meanwhile, the productivities were 1.14 mmol/(L·h) for daidzein, 0.72 mmol/(L·h) for genistein, and 0.19 mmol/(L·h) for glycitein. This is the first report on the application of A. terreus β-glucosidase for converting isoflavone glycosides to their aglycones in soybean products.
Molecularly imprinted polymers as the extracted sorbents of clenbuterol ahead of liquid chromatographic determination
Sovichea Lay, Hai-ning Yu, Bao-xiang Hu, Sheng-rong Shen
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 465-475.   https://doi.org/10.1631/jzus.B1500225
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A pre-treatment methodology for clenbuterol hydrochloride (CLEN) isolation and enrichment in a complex matrix environment was developed through exploiting molecularly imprinted polymers (MIPs). CLEN-imprinted polymers were synthesized by the combined use of ally-β-cyclodextrin (ally-β-CD) and methacrylic acid (MAA), allyl-β-CD and acrylonitrile (AN), and allyl-β-CD and methyl methacrylate (MMA) as the binary functional monomers. MAA-linked allyl-β-CD MIPs (M-MAA) were characterized by Fourier transform-infrared (FT-IR) spectroscopy and a scanning electron microscope (SEM). Based upon the results, M-MAA polymers generally proved to be an excellent selective extraction compared to its references: AN-linked allyl-β-CD MIPs (M-AN) and MMA-linked allyl-β-CD MIPs (M-MMA). M-MAA polymers were eventually chosen to run through a molecularly imprinted solid-phase extraction (MISPE) micro-column to enrich CLEN residues spiked in pig livers. A high recovery was achieved, ranging from 91.03% to 96.76% with relative standard deviation (RSD) ≤4.45%.
Evaluation of the major royal jelly proteins as an alternative to fetal bovine serum in culturing human cell lines
Di Chen, Xiao-xuan Xin, Hao-cheng Qian, Zhang-yin Yu, Li-rong Shen
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 476-483.   https://doi.org/10.1631/jzus.B1500295
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Royal jelly (RJ) is a well-known bioactive substance. It contains large amounts of major royal jelly proteins (MRJPs), which express growth-factor-like activity in several animal and human cell lines. However, the question on whether MRJPs possess growth-factor-like activity on all types of cell cultures remains. In order to determine whether MRJPs can be used as an alternative to fetal bovine serum (FBS) in different types of human cell culture, the proliferation of the complex serum with different ratios of MRJPs/FBS (M/F) was evaluated on five cell lines: 293T, HFL-I, 231, HCT116, and Changliver using MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay. The proliferation activity of the combination of the complex M/F serum with cytokines on the test cell lines was also measured. The results demonstrated that the complex serum with M/F 6/4 possessed the highest proliferation activity similar to or in excess of FBS. However, no activity of complex medium with M/F 6/4 was observed in 231 cells, indicating a selectivity of MRJPs on cell types. Compared with the complex medium with M/F 6/4, the complex medium with M/F 6/4 together with two cytokines, epidermal growth factor (EGF) and insulin-transferrin-selenium (ITS), promoted proliferations of Changliver, 293T, HCT116, and HFL-I by 18.73%‒56.19% (P<0.01). Our findings demonstrate that MRJPs could partially replace FBS in culturing many human cell lines.
Investigation of absorption and scattering characteristics of kiwifruit tissue using a single integrating sphere system
Zhen-huan Fang, Xia-ping Fu, Xue-ming He
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2016, 17(6): 484-492.   https://doi.org/10.1631/jzus.B1500086
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For a quantitative understanding of light interaction with fruit tissue, it is critical to obtain two fundamental parameters: the absorption coefficient and the scattering coefficient of the tissue. This study was to investigate the optical properties of kiwifruit tissue at the wavelength of 632.8 nm. The total reflectance and total transmittance of kiwifruit tissue from three parts (including the flesh part, the seed part, and the seed-base part) were measured using a single integrating sphere system. Based on the measured spectral signals, the absorption coefficient μa and the reduced scattering coefficient μs\' of kiwifruit tissue were calculated using the inverse adding-doubling (IAD) method. Phantoms made from Intralipid 20% and India ink as well as a Biomimic solid phantom were used for system validation. The mean values of μa and μs\' of different parts of the kiwifruit were 0.031–0.308 mm−1 and 0.120–0.946 mm−1, respectively. The results showed significant differences among the μa and μs\' of the three parts of the kiwifruit. The results of this study confirmed the importance of studying the optical properties for a quantitative understanding of light interaction with fruit tissue. Further investigation of fruit optical properties will be extended to a broader spectral region and different kinds of fruits.
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