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Associations of Single Nucleotide Polymorphisms in miR-146a, miR-196a, miR-149 and miR-499 with Colorectal Cancer Susceptibility

  • Du, Wei;Ma, Xue-Lei;Zhao, Chong;Liu, Tao;Du, Yu-Liang;Kong, Wei-Qi;Wei, Ben-Ling;Yu, Jia-Yun;Li, Yan-Yan;Huang, Jing-Wen;Li, Zi-Kang;Liu, Lei
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.2
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    • pp.1047-1055
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    • 2014
  • Background: MicroRNAs (miRNAs) are an abundant class of endogenous small non-coding RNAs of 20-25 nucleotides in length that function as negative gene regulators. MiRNAs play roles in most biological processes, as well as diverse human diseases including cancer. Recently, many studies investigated the association between SNPs in miR-146a rs2910164, miR-196a2 rs11614913, miR-149 rs229283, miR-499 rs3746444 and colorectal cancer (CRC), which results have been inconclusive. Methodology/Principal Findings: PubMed, EMBASE, CNKI databases were searched with the last search updated on November 5, 2013. For miR-196a2 rs11614913, a significantly decreased risk of CRC development was observed under three genetic models (dominant model: OR = 0.848, 95%CI: 0.735-0.979, P = 0.025; recessive model: OR = 0.838, 95%CI: 0.721-0.974, P = 0.021; homozygous model: OR = 0.754, 95%CI: 0.627-0.907, P = 0.003). In the subgroup analyses, miR-$196a2^*T$ variant was associated with a significantly decreased susceptibility of CRC (allele model: OR = 0.839, 95%CI: 0.749-0.940, P = 0.000; dominant model: OR = 0.770, 95%CI: 0.653-0.980, P = 0.002; recessive model: OR = 0.802, 95%CI: 0.685-0.939, P = 0.006; homozygous model: OR = 0.695, 95%CI: 0.570-0.847, P = 0.000). As for miR-149 rs2292832, the two genetic models (recessive model: OR = 1.199, 95% CI 1.028-1.398, P = 0.021; heterozygous model: OR = 1.226, 95% CI 1.039-1.447, P = 0.013) demonstrated increased susceptibility to CRC. On subgroup analysis, significantly increased susceptibility of CRC was found in the genetic models (recessive model: OR = 1.180, 95% CI 1.008-1.382, P = 0.040; heterozygous model: OR = 1.202, 95% CI 1.013-1.425, P = 0.013) in the Asian group. Conclusions: These findings supported that the miR-196a2 rs11614913 and miR-149 rs2292832 polymorphisms may contribute to susceptibility to CRC.

Mapping, Tissue Distribution and Polymorphism of Porcine Retinol Binding Protein Genes (RBP5 and RBP7)

  • Gong, W.H.;Tang, Z.L.;Han, J.L.;Yang, S.L.;Wang, H.;Li, Y.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.11
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    • pp.1544-1550
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    • 2008
  • The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.

Inhibitory Effects of Ssangbohwan on Osteoclast Differentiation and Bone Resorption (쌍보환 추출물의 파골세포 분화 억제와 골 흡수 억제효과)

  • Kim, Seong Joung;Lee, Jeong Ju;Kim, June Hyun;Jo, So Hyun;Park, Min Cheol;Jo, Eun Heui
    • Journal of Acupuncture Research
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    • v.32 no.3
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    • pp.69-81
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    • 2015
  • Purpose : The first purpose of this study is to find out whether the water extract of Rehmanniae Radix Preparat(RRP), Cuscutae Semen(CS) and their combination(Ssangbohwan, SBH) have the effect of suppressing Receptor activator of nuclear factor kappa-B ligand(RANKL)-induced osteoclast differentiation. The second purpose of this study is to find out whether the water extract of RRP, CS and SBH have the effect of inhibiting osteoporosis in an osteoporosis model induced by lipopolysaccharide(LPS). Methods : After promoting differentiation of osteoclasts by treating the RANKL, we observed the effect by the administration of RRP, CS and SBH. In addition, by means of Reverse transcription polymerase chain reaction(RT-PCR), we assayed mRNA expression levels of NFATc1, c-Fos, TRAP and GAPDHS(Glyceraldehyde-3-phosphate dehydrogenase, spermatogeni) from bone marrow macrophages(BMMs). Similarly, the protein expression levels of NFATc1 (Nuclear factor of activated T-cells, cytoplasmic1), C-Fos, MAPKs(Mitogen-activated protein kinases) and ${\beta}$-actin in cell lysates were analyzed by means of Western Blotting. Finally, we determined the anti-osteoporotic effects of RRP, CS and SBH, through the use of Lipopolysaccharide-induced bone-loss mouse. Results : RRP, CS and SBH showed remarkable inhibitive effect on RANKL-treated osteoclast differentiation without cytotoxicity. SBH inhibited the phosphorylation of p38, Jun N-terminal kinases(JNK), and I-${\kappa}B$ and down-regulated the induction of c-Fos and NFATc1 by RANKL. RRP, CS suppressed degradation of I-${\kappa}B$, but it did not affect c-Fos and NFATc1 by RANKL. Lastly, in vivo data showed that RRP and SBH prevented bone erosion by LPS treatment. Conclusions : These results demonstrate SBH can be effective remedy for bone-loss diseases such as osteoporosis.

Inhibitory Effects of Water Extracts of Eucommiae Cortex and Psoraleae Semen Alone and in Combination on Osteoclast Differentiation and Bone

  • Park, Jin Soo;Park, Ga Young;Choi, Han Gyul;Kim, Seong Joung;Kim, June Hyun;park, Min Cheol;Kim, Yun Kyung;Han, Sang Yong;Jo, Eun Heui
    • Journal of Acupuncture Research
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    • v.34 no.2
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    • pp.1-18
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    • 2017
  • Objectives : The purpose of this study was to evaluate the effects of water extracts of Eucommiae cortex (EC), Psoraleae semen (PS), and their combination on receptor activator of nuclear factor-kappa-B ligand (RANKL)-induced osteoclast differentiation. Methods : We assayed the protein expression levels of nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), c-Fos, mitogen-activated protein kinases (MAPKs), and ${\beta}-actin$ in cell lysates using western blotting. Similarly, mRNA expression levels of NFATc1, c-Fos, tartrateresistant acid phosphate (TRAP), and glyceraldehyde-3-phosphate dehydrogenase, spermatogeni (GAPDHS) from bone marrow macrophages (BMMs) were analyzed using reverse transcription-polymerase chain reaction (RT-PCR). Furthermore, we determined the anti-osteoporotic effects of the water extracts of EC, PS, and their combination in a lipopolysaccharide (LPS)-induced bone-loss mouse model. Results : The in vitro data revealed showed that the combination of EC and PS extract showed a more remarkable inhibition of osteoclast differentiation than each herb did alone. The combination downregulated the induction of c-Fos, NFATc1, and TRAP by suppressing the phosphorylation of p38 and c-Jun N-terminal kinases (JNKs) and inhibiting nuclear factor kappa-light-chain-enhancer of activated B cells ($NF-{\kappa}B$). Lastly, the in vivo data showed that PS reduced the LPS-induced bone erosion. Conclusion : The result of this study suggests that EC and PS could be potential therapeutic agents for bone loss diseases such as osteoporosis.

CONSTRUCTION OF HNGF-$\beta$ RECOMBINANT ADENOVIRUS & SCREENING OF ITS EXPRESSION AFTER TRANSFECTION INTO VARIOUS CELL LINES (말초신경재생을 위한 hNGF-$\beta$ recombinant Adenovirus의 제작 및 수종세포주에서 신경성장인자의 발현)

  • Gao, En-Feng;Chung, Hun-Jong;Ahn, Kang-Min;Kim, Yoon-Tae;Park, Hee-Jung;Sung, Mi-Ae;Kim, Nam-Yeol;Yoo, Sang-Bae;Myoung, Hoon;Hwang, Soon-Jung;Kim, Myung-Jin;Kim, Sung-Min;Jang, Jeong-Won;Lee, Jong-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.5
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    • pp.446-456
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    • 2005
  • Nerve growth factor(NGF) has a critical role in peripheral nerve regeneration. The aim of this study is to construct a well-functioning hNGF-$\beta$ recombinat adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with adenovirus mediated hNGF-$\beta$ gene transfection into Schwann cells. First PCR associated cloning of GFP-tagged hNGF-$\beta$ which was ligated into E1/E3 deleted adenoviral vector was performed and tranfected into E. coli to construct hNGF-$\beta$ recombinant adenovirus. After production of recombinat adenovirus in a large scale, its transfection efficiency, expression, and function were evaluated using cell lines or primarily cultured cells of HEK293 cells, Schwann cells, fibroblast(NIH3T3) and myocyte(CRH cells). GFP expression was observed in 90% of infected cells compared to uninfected cells. Total mRNA isolated from hNGF-$\beta$ recombinat adenoviru infected cells showed strong RT-PCR band, however, LacZ recombinant adenovirus infected or uninfected cells did not. NGF quantification by ELISA showed a maximal release of 18.865 +/- 0.31ng/mL at 4th day. PC-12 cells exposed to media with hNGF-$\beta$ recombinant adenovirus infected Schwann cell demonstrated higher levels of differentiation compared with controls. We generated hNGF-$\beta$ recombinant adenovirus and induced over expression of NGF successfully in nonneuronal and neuronal cells. Following these result, it is expected to develop an improved treatment strategy peripheral nerve regeneration using the hNGF-$\beta$ gene transfected cells.

The Comparative Study of Immunomodulatory Effect by Glycyrrhiza New Varieties and Official Compendia (감초 신품종 및 약전 수재감초의 면역조절 효과 비교 연구)

  • Kang, Yun-Mi;Kim, Wonnam;Jin, Jong-Sik;Lee, Jong-Hyun;Chang, Jae Ki;Lee, Jeonghoon;An, Hyo-Jin
    • The Korea Journal of Herbology
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    • v.35 no.6
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    • pp.11-19
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    • 2020
  • Objective : The genus Glycyrrhiza has been used in food and traditional herbal medicine. Glycyrrhiza new varieties Wongam and Sinwongam have been developed by Korea Rural Development Administration and investigated to register on Korean Pharmacopoeia of the Ministry of Food and Drug Safety. The aim of this study is to investigate the immunomodulatory effect of Wongam and Sinwongam comparing with listed Glycyrrhiza species (Glycyrrhiza uralensis Fischer and G. glabra Linne) for evaluations about pharmacological effect of Glycyrrhiza new varieties. Methods : We studied the immunomodulatory effect of Wongam and Sinwongam compared with G. uralensis and G. glabra using THP-1 cell in vitro model. The cells were treated with phorbol 12-myristate 13-acetate (PMA) for differentiation and stimulated with lipopolysaccharides (LPS) to induce immune activation. We analyzed and compared the effects Glycyrrhiza new varieties and listed Glycyrrhiza species using nitric oxide (NO) assay, western blot, and reverse transcription-quantitative polymerase chain reaction analysis. 1) Results : Wongam and Sinwongam showed no cytotoxicity in THP-1 cells. Wongam and Sinwongam, and listed Glycyrrhiza species increased NO production, and cyclooxygenase (COX)-2 expression with or without LPS in differentiated THP-1 macrophages. Furthermore, Wongam and Sinwongam and listed Glycyrrhiza species upregulated the mRNA expressions of T helper type 1 (Th 1)-associated cytokines in LPS-stimulated THP-1 macrophages. Conclusion : These results indicated that Wongam and Sinwongam would have effect of enhancing immune response through the increase of NO and COX-2 expression, and activate Th1-associated cytokines. The findings of this study suggest the wide applicability of Glycyrrhiza new varieties.

Analysis of SNPs in Bovine CSRP3, APOBEC2 and Caveolin Gene Family (소의 CSRP3, APOBEC2, Caveolin 유전자들의 단일염기다형 분석)

  • Bhuiyan, M.S.A.;Yu, S.L.;Kim, K.S.;Yoon, D.;Park, E.W.;Jeon, J.T.;Lee, J.H.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.719-728
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    • 2007
  • The cysteine and glycine rich protein 3 (CSRP3), apolipoprotein B mRNA editing enzyme catalytic polypeptide‐like 2(APOBEC2) and caveolin (CAV) gene family(CAV1, CAV2, CAV3) have been reported to play important roles for carcass and meat quality traits in pig, mouse, human and cattle. As an initial step, we investigated SNPs in these 5 genes among eight different cattle breeds. Eighteen primer pairs were designed from bovine sequence data of NCBI database to amplify the partial gene fragments. Sequencing results revealed 9 SNPs in the coding regions of three caveolin genes, 1 SNP in CSRP3 and 3 SNPs in APOBEC2 gene. All the identified SNPs were confirmed by PCR-RFLP. Also, 9 more intronic SNPs were detected in these genes. However, all identified mutations in the coding region do not change amino acid sequence. Allelic distributions were significantly different for 5 SNPs in CAV2, CAV3, CSRP3 and APOBEC2 genes among the eight different breeds. These results gave some clues about the polymorphisms of these genes among the cattle breeds and will be useful for further searches for identifying association between these SNPs and meat quality traits in cattle.

Effect of Water Extract of Saussureae Radix in RANKL-induced Osteoclast Differentiation (파골세포 분화에 목향 물 추출물의 효과)

  • Lee, Myeung-Su;Kim, Jeong-Joong;Oh, Jae-Min;Choi, Min-Kyu;Song, Mi-Jin;Ahn, Yong-Hwan;Lee, Jeong-Hugh;Jeon, Byung-Hoon;Park, Kie-In;Jang, Sung-Jo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.3
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    • pp.516-520
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    • 2011
  • Osteoporosis is social problem around the world, because fracture of old age may lead to critical medical condition. Osteoclast is a main target for prevention and treatment of osteoporosis due to their responsibility for bone resorption. Saussureae Radix has been known that has gastro-protective, bronchodilatory effect and has a anti-biotic effect. Saussureae Radix has been widely used in Oriental medicine. However, the effect of extract of Saussureae Radix in osteoclast differentiation remains unknown. Thus, we examined the effect of Saussureae Radix in receptor activator of nuclear factor-${\kappa}$B ligand (RANKL)-induced osteoclast differentiation. From the results of our study, Here we found that Saussureae Radix significantly inhibited osteoclast differentiation induced by RANKL. Saussureae Radix suppressed the activation of NF${\kappa}$B in bone marrow macrophages (BMMs) treated with RANKL. Also, Saussureae Radix significantly inhibited the mRNA expression of c-Fos, tartrate-resistant acid phosphatase (TRAP), osteoclast-associated receptor (OSCAR), nuclear factor of activated T cells (NFAT)c1 and cathepsin K in BMMs treated with RANKL. Particularly, Saussureae Radix greatly inhibited the protein expression of c-fos and NFATc1. especially in the case of NFATc1 expression, a master transcription factor of the differentiation of osteoclasts is very important step for osteoclastogenesis. These results demonstrate that Saussureae Radix may be useful treatment option of bone-related disease such as osteoporosis and rheumatoid arthritis.

Inhibition of in Vitro Growth of Three Soil-borne Turfgrass Diseases by Antagonistic Bacteria from Composted Liquid Manure (가축분뇨액비의 길항미생물에 의한 토양전염성 병원균의 생육억제 효과)

  • Ryu, Ju Hyun;Shim, Gyu Yul;Kim, Ki Sun
    • Horticultural Science & Technology
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    • v.32 no.6
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    • pp.879-886
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    • 2014
  • This study was conducted to test in vitro the antagonistic effect of composted liquid manure (CLM) against soil-borne turfgrass pathogenic fungi, Rhizoctonia solani AG-2-2 (IIIB) (brown patch), R. solani AG-2-2 (IV) (large patch), and Sclerotinia homoeocarpa (dollar spot) for environmentally friendly turfgrass management. CLMs were collected from 9 livestock excretion treatment facilities around the country including Gunwi (GW), Hapcheon (HC), Hoengseong (HS), Icheon (IC), Iksan (IS), Muan (MA), Nonsan (NS), and Yeoju (YJ). CLMs of IC, GW, and IS showed s ignificant (p < 0.05) mycelium growth inhibition that was 17.8%, 20.4%, and 48.0% against R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. A t otal of 110 bacterial isolates were obtained from the CLMs that showed antagonistic effects. Among them, 5, 4, and 10 microbe isolates showed promising antifungal activity against mycelium growth of R. solani AG-2-2 (IIIB), R. solani AG-2-2 (IV), and S. homoeocarpa, respectively. The bacterial isolates ICIIIB60, GWIV70, and ISSH20 effectively inhibited the mycelial growth of three soil-borne turfgrass pathogens. Selected bacterial isolates were identified as Alcaligenes sp., Bacillus licheniformis Ab2, and B. subtilis C7-3 through 16s rDNA gene sequence analysis. Among 5 fungicides, the most compatible fungicide with ICIIIB60, GWIV70, and ISSH20 was tebuconazol, toclofos-methyl and toclofos-methyl, respectively. These findings suggested that CLMs could be effectively used not only as organic liquid fertilizer sources but also as biological control agents for soil-borne turfgrass diseases such as brown patch, large patch, and dollar spot.

Isolation of CONSTANS as a TGA4/OBF4 Interacting Protein

  • Song, Young Hun;Song, Na Young;Shin, Su Young;Kim, Hye Jin;Yun, Dae-Jin;Lim, Chae Oh;Lee, Sang Yeol;Kang, Kyu Young;Hong, Jong Chan
    • Molecules and Cells
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    • v.25 no.4
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    • pp.559-565
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    • 2008
  • Members of the TGA family of basic domain/leucine zipper transcription factors regulate defense genes through physical interaction with NON-EXPRESSOR OF PR1 (NPR1). Of the seven TGA family members, TGA4/octopine synthase (ocs)-element-binding factor 4 (OBF4) is the least understood. Here we present evidence for a novel function of OBF4 as a regulator of flowering. We identified CONSTANS (CO), a positive regulator of floral induction, as an OBF4-interacting protein, in a yeast two-hybrid library screen. OBF4 interacts with the B-box region of CO. The abundance of OBF4 mRNA cycles with a 24 h rhythm under both long-day (LD) and short-day (SD) conditions, with significantly higher levels during the night than during the day. Electrophoretic mobility shift assays revealed that OBF4 binds to the promoter of the FLOWERING LOCUS T (FT) gene, a direct target of CO. We also found that, like CO and FT, an OBF4:GUS construct was prominently expressed in the vascular tissues of leaf, indicating that OBF4 can regulate FT expression through the formation of a protein complex with CO. Taken together, our results suggest that OBF4 may act as a link between defense responses and flowering.