• Title/Summary/Keyword: Chinese radicals

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Soluble Expression of a Human MnSOD and Hirudin Fusion Protein in Escherichia coli, and Its Effects on Metastasis and Invasion of 95-D Cells

  • Yi, Shanze;Niu, Dewei;Bai, Fang;Li, Shuaiguang;Huang, Luyuan;He, Wenyan;Prasad, Anand;Czachor, Alexander;Tan, Lee Charles;Kolliputi, Narasaiah;Wang, Feng
    • Journal of Microbiology and Biotechnology
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    • v.26 no.11
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    • pp.1881-1890
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    • 2016
  • Manganese superoxide dismutase (MnSOD) is a vital enzyme that protects cells from free radicals through eliminating superoxide radicals ($O^{2-}$). Hirudin, a kind of small active peptide molecule, is one of the strongest anticoagulants that can effectively cure thrombus diseases. In this study, we fused Hirudin to the C terminus of human MnSOD with the GGGGS linker to generate a novel dual-feature fusion protein, denoted as hMnSOD-Hirudin. The hMnSOD-Hirudin gene fragment was cloned into the pET15b (SmaI, CIAP) vector, forming a recombinant pET15b-hMnSOD-Hirudin plasmid, and then was transferred into Escherichia coli strain Rosetta-gami for expression. SDS-PAGE was used to detect the fusion protein, which was expected to be about 30 kDa upon IPTG induction. Furthermore, the hMnSOD-Hirudin protein was heavily detected as a soluble form in the supernatant. The purification rate observed after Ni NTA affinity chromatography was above 95%. The hMnSOD-Hirudin protein yield reached 67.25 mg per liter of bacterial culture. The identity of the purified protein was confirmed by western blotting. The hMnSOD-Hirudin protein activity assay evinced that the antioxidation activity of the hMnSOD-Hirudin protein obtained was $2,444.0{\pm}96.0U/mg$, and the anticoagulant activity of the hMnSOD-Hirudin protein was $599.0{\pm}35.0ATU/mg$. In addition, in vitro bioactivity assay showed that the hMnSOD-Hirudin protein had no or little cytotoxicity in H9c2, HK-2, and H9 (human $CD_4{^+}$, T cell) cell lines. Transwell migration assay and invasion assay showed that the hMnSOD-Hirudin protein could suppress human lung cancer 95-D cell metastasis and invasion in vitro.

Antioxidant Activities According To Peeling and Cultivated Years of Astragalus membranaceus Roots (황기(Astragalus membranaceus)의 박피 유무와 재배 년 수에 따른 항산화 활성 연구)

  • Goh, Eun-Jeong;Seong, Eun-Soo;Lee, Jae-Geun;Na, Jong-Kuk;Lim, Jung-Dae;Kim, Myong-Jo;Kim, Na-Young;Lee, Gwi-Hyun;Seo, Jung-Sik;Cheoi, Dae-Sung;Chung, Ill-Min;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.17 no.4
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    • pp.233-237
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    • 2009
  • Astragalus membranaceus has a long history of medicinal use in Chinese herbal medicine. It has been shown to have immunostimulant, tonic, antioxidant, antiperspirant, diuretic, anti-diabetic, expectorant properties, and a supplementary medicine during cancer therapy. In this study, we investigated the effect of anti-oxidation of Astragalus membranaceus root extract. The anti-oxidative activities of water, 80% methanol, and 100% methanol extracts from Astragalus membranaceus were analyzed by DPPH free radical scavenging activity, Superoxide dismutase-like activity, reducing power, and crude ash. The water extract demonstrated to be more effective than methanol extract for a DPPH radicals scavenging activities and reducing power. Superoxide dismutase-like activity showed higher efficiency in 80% methanol extract. Our results indicate that Astragalus membranaceus extracts could be used as a source of antioxidant ingredients in the food industry.

Protective effects of extracts from six local strains of Pyropia yezoensis against oxidative damage in vitro and in zebrafish model

  • Dai, Yu-Lin;Kim, Gwang Hoon;Kang, Min-Cheol;Jeon, You-Jin
    • ALGAE
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    • v.35 no.2
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    • pp.189-200
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    • 2020
  • Pyropia yezoensis has been used as functional food in East Asia, especially in Korea and Japan, for more than five hundred years. This study aims to evaluate the antioxidant effect of polyphenols and proteins-rich extracts from P. yezoensis (PPPs) against 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced oxidative cell damage. Among six Korean local strains obtained from Jinhae (JiH), Haenam (HN), Jangheung (JaH), Jindo (JD), Wando (WD), and Sinan (SA) areas, the extracts of P. yezoensis from SA and JD are relatively higher in polyphenols and proteins contents. SA showed the lowest IC50 scavenging activities against 1,1-diphenyl-2-picryl-hydrazyl and alkyl radicals and displayed protective effects against reactive oxygen species (ROS) in AAPH-induced Vero cells. Especially, the PPPs extracts from SA and JD showed protective activities against AAPH-induced apoptosis, as observed by nuclear staining with Hoechst 33342. Furthermore, in vivo studies of the SA extract in zebrafish showed significantly reduced ROS generation, lipid peroxidation, and cell damage. This is the first study, to our knowledge, to evaluate the antioxidant bioactivity of PPP in the Korean Peninsula using a zebrafish model. Due to SA and JD both located in the west coast of Korea, we deduced that the chemical content of the different PPP extracts was mildly influenced by their geographic location, and this alga has potential of protective activity against AAPH-induced ROS both in vitro and in vivo.

Antioxidant Properties in Water and 70% Ethanol Extracts of Houttuynia Cordata Thunb (어성초의 추출물에 따른 항산화력 연구)

  • Kim, Hyung Soo;Kim, Mi-Ja;Cheong, Chul;Kang, Soon Ah
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.10
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    • pp.5091-5096
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    • 2013
  • Houttuynia cordata Thunb is a famous traditional Chinese medicine. Also it has rich in functional and bioactive substances with outstanding immunopotentiation, anti-cancer, antimicrobial, and anti-aging effects. This study was evaluated water and 70% ethanol extracts of Houttuynia cordata Thunb using in vitro system antioxidant properties methods. The scavenging DPPH and ABTS radicals of 2 mg/mL 70% ethanol extract were 91.8 and 54.8%, respectively. Also, The FRAP assay of this showed increase in activity. In results, the antioxidant properties of 70% ethanol extract increased significantly compared to water extract(p < 0.05). Therefore, 70% ethanol extract of Houttuynia cordata Thunb may contribute to antioxidant property.

Enhanced photo-Fenton degradation of tetracycline using TiO2-coated α-Fe2O3 core-shell heterojunction

  • Zheng, Xiaogang;Fu, Wendi;Kang, Fuyan;Peng, Hao;Wen, Jing
    • Journal of Industrial and Engineering Chemistry
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    • v.68
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    • pp.14-23
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    • 2018
  • $TiO_2-coated$ cubic ${\alpha}-Fe_2O_3$ with mostly exposed (012) and (101) facets (${\alpha}-Fe_2O_3@TiO_2$) was fabricated using a hydrothermal route for the photo-Fenton degradation of tetracycline under visible light irradiation. $TiO_2$ coating could greatly affect the photocatalytic activity of ${\alpha}-Fe_2O_3@TiO_2$. Compared with cubic ${\alpha}-Fe_2O_3$ alone for photodegradation of tetracycline, ${\alpha}-Fe_2O_3@TiO_2$ with $TiO_2$ shell of around 15 nm exhibited higher removal efficiency of tetracycline in photo-Fenton system, and its durability was slightly affected after five cycle times under same conditions. It is ascribed to the well-matched interface between cubic ${\alpha}-Fe_2O_3$ core and $TiO_2$ shell, leading to the broadened light-absorption and the efficient separation of photo-generated electon-hole pairs. The $^{\bullet}OH$ radicals were main responsible for the advanced photocatalytic performance of ${\alpha}-Fe_2O_3@TiO_2$ in visible-light driven degradation of tetracycline.

Antioxidant Activity of Methanol Extracts from the Genus Lespedeza (싸리속 식물 메탄올 추출물의 항산화 활성)

  • Kim, Sang-Min;Jung, Yu-Jin;Pan, Cheol-Ho;Um, Byung-Hun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.5
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    • pp.769-775
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    • 2010
  • The genus Lespedeza belonging to Leguminosae is an annual or perennial herb, which has been used as a Chinese herbal medicine. Antioxidant activity on DPPH ((1,1)-diphenyl-2-picrylhydrazyl) and ABTS (2,2'-azino-bis(3-ethylenebenzothiazoline-6-sulfonic acid) radicals and the contents of total polyphenol and flavonoid were analyzed with 12 methanol extracts from six Lespedeza genus plants with different plant parts. The highest content of total polyphenol was detected in the leaves of Lespedeza$\times$robusta (194.6 mg GAE/g), while the highest content of total flavonoid existed in the aerial part of Lespedeza thunbergii var. intermedia (47.9 mg QE/g). Generally, the amounts of polyphenol and flavonoid compounds were higher in the leaves than in the stems. The $SC_{50}$ value of DPPH radical scavenging activity ranged from 10.16 to 90.94 ppm, of which the strongest activity was determined in the leaves of L. robusta. Most ABTS radical scavenging activities from the investigated methanol extracts were higher than that of L-ascorbic acid, implying the excellent antioxidant activity. The radical scavenging activity in this study showed high correlation with the amount of total polyphenol rather than that of total flavonoid. These data suggest that the methanol extracts from these Lespedeza spp. could be potential candidates for natural antioxidants.

Changes in the quality characteristics of kimchi broths and growth of the isolated strains due to blue light emitting diode irradiation (Blue LED (Light emitting diode) 조사에 의한 배추김치와 백김치 발효액의 품질 특성 및 분리 균주의 생육 변화)

  • Oh, Yeong Ji;Hong, Jungil
    • Korean Journal of Food Science and Technology
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    • v.52 no.5
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    • pp.538-545
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    • 2020
  • Kimchi is a widely consumed traditional Korean food, and its probiotic properties have received great attention. In this study, changes in the quality characteristics of fermentation broths obtained from two types of Chinese cabbage kimchi (red with red pepper and white without red pepper) were assessed after the administration of blue light emitting diode (BLED) irradiation at 4℃; characteristics assessed included acidity, chromaticity, antioxidant activity, and growth modulation of isolated microorganisms. The pH of the white kimchi (WK) broth decreased with time; the decrease was delayed significantly under BLED irradiation (p<0.05). BLED irradiation decreased the L (lightness) and b (yellowness) values and increased a (redness) in WK, whereas the a and b values of the red kimchi (RK) broth increased with BLED irradiation. Growth stimulation of lactic acid bacteria by BLED irradiation was observed in both WK and RK. The numbers of yeast and mold were also increased in RK (p<0.05), but not in WK. There was no change observed in the scavenging activities against ABTS (2,2'-azido-bis(3-ethylbenzthiazoline-6-sulfonic acid) radicals in both kimchi broths after BLED treatment. The results of this study indicated that BLED irradiation could modulate the fermentation process and the quality characteristics of kimchi during storage.

Protective Effects of a Herb, Artemisia capillaris, Against Radiation-induced DNA Damage (방사선 유도 DNA 손상에 대한 인진쑥의 방어효과)

  • Jo, Sung-Kee;Oh, Heon;Cheon, Eui-Hyun;Jeong, U-Hee;Cho, Nam-Jeong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.1
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    • pp.22-27
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    • 2004
  • In the present study, the protective effects of Artemisia capillaris (AC) on the DNA damage induced by $^{60}$ Co ${\gamma}$-rays were evaluated using alkaline single-cell gel electrophoresis (SCGE, comet assay) in the mouse peripheral lymphocytes and micronuclei (MN) formation test in the Chinese hamster ovary (CHO) cells. We also investigated the effect of AC on 8-hydroxy-2'-deoxyguanosine (8-OHdG) formation in the mouse liver and thymus exposed to ${\gamma}$-ray, The tail moment and the frequency of MN, which were markers of DNA damage in the SCGE and MN formation test, were decreased in the groups treated with AC extract before exposure to 200 cGy of ${\gamma}$-ray. We also observed its activities, lowering 8-OHdG level, an index of oxidative DNA damage, in the groups treated with AC extract before whole body ${\gamma}$-irradiation (800 cGy). It is plausible that scavenging of free radicals by AC may have played an important role in providing the protection against the radiation-induced damage to the DNA. These results indicated that AC protects the DNA damage induced by ${\gamma}$-rays and might be a useful radioprotector, especially since it is a relatively nontoxic product.

Photoprotection and Anti-inflammatory Effects of Chinese Medical Plants (약용식물추출물의 광보호 효과와 항염증 효과 연구)

  • Jin-Hwa, Kim;Sung-Min, Park;Gwan-Sub, Sim;Bum-Chun , Lee;Hyeong-Bae, Pyo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.227-233
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    • 2004
  • Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.