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Kwon, J. H.,Kim, J. H.,Lee, D. h.,Cho, H.,Hwang, S. Y.,Yuk, S. S.,Erdene-Ochir, T. O.,Noh, J. Y.,Hong, W. T.,Jeong, J. H. Springer Science + Business Media 2016 BioChip Journal Vol.10 No.3
<P>Highly pathogenic avian influenza (HPAI) viruses cause economic losses in the poultry industry and pose a severe threat to human health. Rapid and accurate diagnostic methods are important because they can help prevent further spread of the virus and reduce the time required for eradication of the virus. We developed a low-density microarray for the rapid detection and identification of avian influenza virus subtypes H5, H7, and H9 and their pathotypes in a previous study. In the present study, we report the development of updated probe sets and evaluation of the diagnostic microarray using H5N8 clade 2.3.4.4 HPAI viruses including clinical samples, without the need for egg propagation. Cy3-labeled DNA targets were obtained by reverse transcription polymerase chain reaction using Cy3-labeled universal primers, and labeled amplicons were hybridized to the microarray. All positive samples from RT-PCR showed H5-specific and highly pathogenic pattern in the microarray, without purification of PCR products. Furthermore, it allowed for specific detection of the subtype and pathotype from low DNA concentration samples that did not allow direct sequence analysis. Therefore, this diagnostic microarray has enormous potential for the rapid subtyping and pathotyping from clinical samples without the need for culture.</P>
Shin, M.J.,Kim, D.W.,Jo, H.S.,Cho, S.B.,Park, J.H.,Lee, C.H.,Yeo, E.J.,Choi, Y.J.,Kim, J.A.,Hwang, J.S.,Sohn, E.J.,Jeong, J.H.,Kim, D.S.,Kwon, H.Y.,Cho, Y.J.,Lee, K.,Han, K.H.,Park, J.,Eum, W.S.,Choi, Pergamon ; Elsevier Science Ltd 2016 FREE RADICAL BIOLOGY AND MEDICINE Vol.97 No.-
Proline rich Akt substrate (PRAS40) is a component of mammalian target of rapamycin complex 1 (mTORC1) and is known to play an important role against reactive oxygen species-induced cell death. However, the precise function of PRAS40 in ischemia remains unclear. Thus, we investigated whether Tat-PRAS40, a cell-permeable fusion protein, has a protective function against oxidative stress-induced hippocampal neuronal (HT-22) cell death in an animal model of ischemia. We showed that Tat-PRAS40 transduced into HT-22 cells, and significantly protected against cell death by reducing the levels of H<SUB>2</SUB>O<SUB>2</SUB> and derived reactive species, and DNA fragmentation as well as via the regulation of Bcl-2, Bax, and caspase 3 expression levels in H<SUB>2</SUB>O<SUB>2</SUB> treated cells. Also, we showed that transduced Tat-PARS40 protein markedly increased phosphorylated RRAS40 expression levels and 14-3-3σ complex via the Akt signaling pathway. In an animal ischemia model, Tat-PRAS40 effectively transduced into the hippocampus in animal brain and significantly protected against neuronal cell death in the CA1 region. We showed that Tat-PRAS40 protein effectively transduced into hippocampal neuronal cells and markedly protected against neuronal cell damage. Therefore, we suggest that Tat-PRAS40 protein may be used as a therapeutic protein for ischemia and oxidative stress-induced brain disorders.
Hwang, H.J.,Jung, T.W.,Ryu, J.Y.,Hong, H.C.,Choi, H.Y.,Seo, J.A.,Kim, S.G.,Kim, N.H.,Choi, K.M.,Choi, D.S.,Baik, S.H.,Yoo, H.J. North-Holland 2014 Molecular and cellular endocrinology Vol.392 No.1
The direct effects of dipeptidyl peptidase-IV (DPP-IV) inhibitors on endoplasmic reticulum (ER) stress-induced apoptosis and inflammation in cardiomyocytes have not been elucidated. H9c2 cell viability, which was reduced by tunicamycin, was increased after DPP-IV inhibitor gemigliptin treatment. Gemigliptin significantly decreased the tunicamycin-mediated increase in glucose regulated protein 78 (GRP78) expression and ER stress-mediated signaling molecules such as protein kinase RNA-like endoplasmic reticulum kinase (PERK)/C-EBP homologous protein (CHOP) and inositol-requiring enzyme 1α (IRE1α)/c-Jun N-terminal kinase (JNK)-p38. Furthermore, gemigliptin effectively induced Akt phosphorylation in a dose-dependent manner. Using flow cytometry and Hoechst staining, we showed that treatment with Akt inhibitor significantly blocked the anti-apoptotic effects mediated by gemigliptin. The reduction in tunicamycin-induced GRP78 level and PERK/CHOP pathway activity by gemigliptin was reversed after treatment with Akt inhibitor. In conclusion, gemigliptin effectively inhibited ER stress-induced apoptosis and inflammation in cardiomyocytes via Akt/PERK/CHOP and IRE1α/JNK-p38 pathways, suggesting its direct protective role in cardiovascular diseases.
Hwang, S.M.,Sung, K.,Choi, J.H.,Kim, W.,Joo, J.,Lim, J.H.,Kim, C.J.,Park, Y.S.,Kim, D.H. North-Holland 2010 Physica. C, Superconductivity Vol.470 No.20
We selected polyacrylonitrile (PAN, -[C<SUB>3</SUB>H<SUB>3</SUB>N]-) as an O-free organic dopant and fabricated C-doped MgB<SUB>2</SUB> wires by in situ and powder-in-tube techniques. 0-5 wt.% PAN powders were uniformly mixed with B powder using a liquid mixing method. The precursor powders were mixed with Mg powder, filled into Fe tubes, and then drawn into wires. Sintering was performed at 900<SUP>o</SUP>C for 1h in a flowing Ar gas. The PAN doping decreased the critical temperature (T<SUB>c</SUB>) and a-axis lattice parameter, but significantly improved the critical current density (J<SUB>c</SUB>) in high fields, upper critical field (H<SUB>c2</SUB>), and irreversibility field (H<SUB>irr</SUB>) performances. These results are attributed to the replacement of B sites with C by the PAN doping. Furthermore, as expected, the MgO amount did not increase as the doping content increased. The J<SUB>c</SUB> of the PAN-doped MgB<SUB>2</SUB> wires was more than one order of magnitude higher than that of the undoped MgB<SUB>2</SUB> wire at 5K and 6.6T (1.46-3.82kA/cm<SUP>2</SUP> vs. 0.11kA/cm<SUP>2</SUP>).
Joung, H.J.,Kim, Y.S.,Hwang, J.W.,Han, Y.K.,Jeong, J.H.,Lee, J.S.,Moon, S.H.,Jeon, B.T.,Park, P.J. Academic Press 2014 FISH AND SHELLFISH IMMUNOLOGY Vol.38 No.1
In this study, Haliotis discus hannai (H. discus hannai) fermentation was attempted with Cordyceps militaris (C. militaris) mycelia using a solid culture. We tried to ferment H. discus hannai to determine the optimal conditions fermentation with regards to its anti-inflammatory effects. The extracts of H. discus hannai fermented with C. militaris mycelia (HFCM-5) showed higher nitric oxide inhibitory effects than H. discus hannai and C. militaris alone. HFCM-5 also decreased pro-inflammatory cytokines, TNF-α and IL-6 in a dose-dependent manner. HFCM-5 did not affect the MyD88-dependent pathway, but decreased phosphorylation of IRF3 and STAT1 which are involved in TRIF-dependent pathway. Taken together, our results suggest that HFCM-5 exerts its anti-inflammatory effects via TRIF signaling pathway and could potentially be used as a functional food in the regulation of inflammation.
Park, H.J.,Jeong, J.M.,Bae, J.S.,Kim, J.W.,An, C.M.,Min, B.H.,Kim, S.Y.,Myeong, J.I.,Hwang, H.K.,Park, C.I. Pergamon Press ; Elsevier Science 2016 DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY Vol.65 No.-
<P>A new lily-type lectin RbLTL was identified from rock bream (Oplegnathus fasciatus) and its expression analysed. In this study, a new lily-type lectin gene (RbLTL) was cloned from rock bream using expressed sequence tag (EST) analysis. The full-length RbLTL cDNA was encoding a 117-amino acid protein. The deduced amino acid sequence of RbLTL contained all of the conserved features crucial for its fundamental structure, including B-lectin domain and three D-mannose binding sites. RbLTL mRNA was predominately expressed in the gills, with reduced expression noted in intestine tissue. Expression analysis of time series sampled fertilized eggs revealed that expression gradually increased 1, 3, 12, and 24 h: However, expression decreased at 36 h. RbLTL expression was differentially up-regulated in rock bream gills challenged with Streptococcus iniae, Edwardsiella tarda and RSIV. Our results revealed that novel rock bream lily-type lectin may be an important molecule involved in pattern recognition and pathogen elimination in the innate immunity of rock bream. (C) 2016 Elsevier Ltd. All rights reserved.</P>
Escherichia coli-based expression of functional novel DNA-binding histone H1 from Carassius auratus
Wei, Q.,Jung, H.J.,Hwang, D.S.,Hwang, B.H.,Gim, Y.,Cha, H.J. IPC Science and Technology Press ; Elsevier Scienc 2007 Enzyme and microbial technology Vol.40 No.6
Histones are DNA-binding proteins that assist in DNA packaging and protection. Here, we, for the first time, cloned a novel histone H1 cDNA (638bp) from the goldfish, Carassius auratus. Sequencing revealed that this histone H1 shared 68.1% amino acid identity and 73.9% similarity with that from the rainbow trout, Salmo gairdneri. We successfully expressed a full-length recombinant version (∼20kDa) and a truncated C-terminal fragment (∼6kDa; 61 amino acids) of this histone H1 as a partially soluble form using a maltose binding protein (MBP) fusion strategy in an Escherichia coli expression system. Our results revealed that both these recombinant histone H1 versions had DNA binding and protection functions, and MBP fusion system was an effective way to produce biological functional recombinant histone proteins in E. coli. This novel recombinant histone H1 protein and/or its C-terminal peptide could be used as potential mediators for efficacious gene delivery.
An exploration of the antioxidant effects of garlic saponins in mouse-derived C2C12 myoblasts
Kang, J. S.,Kim, S. O.,Kim, G.-Y.,Hwang, H. J.,Kim, B. W.,Chang, Y.-C.,Kim, W.-J.,Kim, C. M.,Yoo, Y. H.,Choi, Y. H. Spandidos Publications 2016 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.37 No.1
<P>In this study, we aimed to confirm the protective effects of garlic saponins against oxidative stress-induced cellular damage and to further elucidate the underlying mechanisms in mouse-derived C2C12 myoblasts. Relative cell viability was determined by 3-(4.5-dimethylthiazol-2-yl)-2.5 diphenyltetrazolium bromide assay. Comet assay was used to measure DNA damage and oxidative stress was determined using 2 ',7 '-dichlorofluorescein diacetate to measure intracellular reactive oxygen species (ROS) generation. Western blot analysis and small interfering RNA (siRNA)-based knockdown were used in order to investigate the possible molecular mechanisms. Our results revealed that garlic saponins prevented hydrogen peroxide (H2O2)-induced growth inhibition and exhibited scavenging activity against intracellular ROS. We also observed that garlic saponins prevented H2O2-induced comet tail formation and decreased the phosphorylation levels of gamma H2AX expression, suggesting that they can prevent H2O2-induced DNA damage. In addition, garlic saponins increased the levels of heme oxygenase-1 (HO-1), a potent antioxidant enzyme associated with the induction and phosphorylation of nuclear factor erythroid 2-related factor 2 (Nrf2) and the translocation of Nrf2 from the cytosol into the nucleus. However, the protective effects of garlic saponins on H2O2-induced ROS generation and growth inhibition were significantly reduced by zinc protoporphyrin IX, an HO-1 competitive inhibitor. In addition, the potential of garlic saponins to mediate HO-1 induction and protect against H2O2-mediated growth inhibition was adversely affected by transient transfection with Nrf2-specific siRNA. Garlic saponins activated extracellular signal-regulated kinase (ERK) signaling, whereas a specific ERK inhibitor was able to inhibit HO-1 upregulation, as well as Nrf2 induction and phosphorylation. Taken together, the findings of our study suggest that garlic saponins activate the Nrf2/HO-1 pathway by enabling ERK to contribute to the induction of phase II antioxidant and detoxifying enzymes, including HO-1 in C2C12 cells.</P>
Kim, J.W.,Park, C.I.,Hwang, S.D.,Jeong, J.M.,Kim, K.H.,Kim, D.H.,Shim, S.H. Academic Press 2013 Fish & shellfish immunology Vol.35 No.1
Cathepsins are lysosomal cysteine proteases belonging to the papain family, whose members play important roles in normal metabolism for the maintenance of cellular homeostasis. Rock bream (Oplegnathus fasciatus) cathepsin H (RbCTSH) cDNAs were identified by expressed sequence tag analysis of a lipopolysaccharide-stimulated rock bream liver cDNA library. The full-length RbCTSH cDNA (1326 bp) contained an open reading frame of 978 bp encoding 325 amino acids. The presence of an ERFNIN-like motif was predicted in the propeptide region of RbCTSH. Furthermore, multiple alignments showed that the EPQNCSAT region was well conserved among other cathepsin H sequences. Phylogenetic analysis revealed that RbCTSH is most closely related to Nile tilapia cathepsin H. RbCTSH was expressed significantly in the intestine, spleen, head kidney and stomach. RbCTSH mRNA expression was also examined in several tissues under conditions of bacterial and viral challenge. All examined tissues of fish infected with Edwardsiella tarda, Streptococcus iniae and red sea bream iridovirus (RSIV) showed significant increases in RbCTSH expression compared to the control. In the kidney and spleen, RbCTSH mRNA expression was upregulated markedly following infection with bacterial pathogens. These findings indicate that RbCTSH plays an important role in the innate immune response of rock bream. Furthermore, these results provide important information for the identification of other cathepsin H genes in various fish species.