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Effect of Atractylodis Rhizoma Alba on Osteoclast Formation (백출의 파골세포 분화에 미치는 영향)

  • Park, Sung-Tae;Lee, Myeung-Su;Jeon, Byung-Hun;Park, Kie-In;Oh, Jae-Min
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.1
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    • pp.109-114
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    • 2011
  • Atractylodis Rhizoma Alba is commonly used herbal medicine and it has been known that has immuno-regualtory effects and anti-cancer effects. The inhibition of osteoclastogenesis is essential for the prevention and treatment of osteoporosis. The aim of this study was to evaluate the effects of Atractylodis Rhizoma Alba on osteoclast differentiation in vitro and on resorbing activity of osteoclast. Osteoclast formation was evaluated in bone marrow cells (BMC) in the presence or absence of Atractylodis Rhizoma Alba. The expression of c-fos, tartrate-resistant acid phosphatase (TRAP), OSCAR, DC-STAMP, cathepsin K, MafB and NFATc1 mRNA in osteoclast precursor were assessed by RT-PCR. The levels of TNF receptor-associated factor-6 (TRAF-6), c-fos and NFATc1 protein were assessed by Western blot analysis. Also the correlation with MAPKs and NF-${\kappa}B$ pathways were measured by using Western blot analysis. With bone resorption study, I tried to evaluate the inhibitory effects of Atractylodis Rhizoma Alba on mature osteoclast function. Atractylodis Rhizoma Alba inhibited the RANKL induced osteoclastic differentiation from bone marrow macrophage in a dose dependant manner without cellular toxicity. Gene expression of c-fos and NFATc1 was significantly down regulated with Atractylodis Rhizoma Alba treatment. Atractylodis Rhizoma Alba markedly inhibited the RANKL-induced osteoclastogenesis through suppression of nuclear factor kappa b (NF-${\kappa}B$) pathway, down stream pathway of p38, ERK and JNK pathway. Taken together, I concluded that Atractylodis Rhizoma Alba have beneficial effect on osteoporosis by inhibition of osteoclast differentiation and by inhibition of functioning osteoclast. Thus I expect that Atractylodis Rhizoma Alba could be a treatment option for osteoporosis.

Identification of a SNP in Cattle HGD Gene with its Effect on Economic Trait in Hanwoo (한우 HGD 유전자내 변이지역과 경제형질간의 연관성 분석)

  • Han, Jung-Min;Kong, Hong Sik
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1168-1173
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    • 2014
  • The homogentisate 1,2-dioxygenase (HGD) gene, which consists of 14 exons and spans approximately 42630bp on Bos taurus autosome 1 (BTA 1), is one of the six enzymes required for catabolism of the aromatic amino acids tyrosine and phenylalanine. It has been reported that BTA1 harbors quantitative trait loci that effect marbling score (MS), carcass weight (CW), and longissimus muscle area (LMA) in cattle. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) in the HGD gene and to analyze their association with economic traits in Korean cattle (Hanwoo). Genetic polymorphisms were screened by direct sequencing, which detected 10 SNPs (T11187C, T11301A, T11398G, G29833A, G34256T, G34257C, T34284C, T42333G, T42348C, and T42468C). Six polymorphic sites were selected for genotyping, and economic traits were analyzed using a general linear model in Korean cattle (n=90). The observed genotype frequencies for G34256T were 0.5843(GG), 0.3708(GT), and 0.0449(TT). In addition, 0.3596(GG), 0.3708(GC), and 0.2697(CC) were observed for the G34257C mutation. Statistical association analysis revealed that G34256T polymorphisms were significantly associated with MS, and G34257C polymorphisms were significantly associated with MS and LMA (p<0.05). Further study is needed in order to use the genetic variant as a marker for marker-assisted selection in Korean cattle.

Evaluation of Genetic Diversity and Population Structure Analysis among Germplasm of Agaricus bisporus by SSR Markers

  • An, Hyejin;Lee, Hwa-Yong;Shin, Hyeran;Bang, Jun Hyoung;Han, Seahee;Oh, Youn-Lee;Jang, Kab-Yeul;Cho, Hyunwoo;Hyun, Tae Kyung;Sung, Jwakyung;So, Yoon-Sup;Jo, Ick-Hyun;Chung, Jong-Wook
    • Mycobiology
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    • v.49 no.4
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    • pp.376-384
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    • 2021
  • Agaricus bisporus is a popular edible mushroom that is cultivated worldwide. Due to its secondary homothallic nature, cultivated A. bisporus strains have low genetic diversity, and breeding novel strains is challenging. The aim of this study was to investigate the genetic diversity and population structure of globally collected A. bisporus strains using simple sequence repeat (SSR) markers. Agaricus bisporus strains were divided based on genetic distance-based groups and model-based subpopulations. The major allele frequency (MAF), number of genotypes (NG), number of alleles (NA), observed heterozygosity (HO), expected heterozygosity (HE), and polymorphic information content (PIC) were calculated, and genetic distance, population structure, genetic differentiation, and Hardy-Weinberg equilibrium (HWE) were assessed. Strains were divided into two groups by distance-based analysis and into three subpopulations by model-based analysis. Strains in subpopulations POP A and POP B were included in Group I, and strains in subpopulation POP C were included in Group II. Genetic differentiation between strains was 99%. Marker AB-gSSR-1057 in Group II and subpopulation POP C was confirmed to be in HWE. These results will enhance A. bisporus breeding programs and support the protection of genetic resources.

Comprehensive Bioinformation Analysis of the MRNA Profile of Fascin Knockdown in Esophageal Squamous Cell Carcinoma

  • Wu, Bing-Li;Luo, Lie-Wei;Li, Chun-Quan;Xie, Jian-Jun;Du, Ze-Peng;Wu, Jian-Yi;Zhang, Pi-Xian;Xu, Li-Yan;Li, En-Min
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7221-7227
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    • 2013
  • Background: Fascin, an actin-bundling protein forming actin bundles including filopodia and stress fibers, is overexpressed in multiple human epithelial cancers including esophageal squamous cell carcinoma (ESCC). Previously we conducted a microarray experiment to analyze fascin knockdown by RNAi in ESCC. Method: In this study, the differentially expressed genes from mRNA expression profilomg of fascin knockdown were analyzed by multiple bioinformatics methods for a comprehensive understanding of the role of fascin. Results: Gene Ontology enrichment found terms associated with cytoskeleton organization, including cell adhesion, actin filament binding and actin cytoskeleton, which might be related to fascin function. Except GO categories, the differentially expressed genes were annotated by 45 functional categories from the Functional Annotation Chart of DAVID. Subpathway analysis showed thirty-nine pathways were disturbed by the differentially expressed genes, providing more detailed information than traditional pathway enrichment analysis. Two subpathways derivated from regulation of the actin cytoskeleton were shown. Promoter analysis results indicated distinguishing sequence patterns and transcription factors in response to the co-expression of downregulated or upregulated differentially expressed genes. MNB1A, c-ETS, GATA2 and Prrx2 potentially regulate the transcription of the downregulated gene set, while Arnt-Ahr, ZNF42, Ubx and TCF11-MafG might co-regulate the upregulated genes. Conclusions: This multiple bioinformatic analysis helps provide a comprehensive understanding of the roles of fascin after its knockdown in ESCC.

The Stimulatory Effect of P2Y6 Receptor Antagonist on RANKL-induced Osteoclastogenesis (P2Y6 수용체 길항제의 파골세포 분화 촉진 효과 규명)

  • Noh, A Long Sae Mi;Moon, Miran;Yim, Mijung
    • YAKHAK HOEJI
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    • v.59 no.5
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    • pp.207-214
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    • 2015
  • P2Y receptors, a type of P2 receptor family, are G-protein coupled receptors and 8 subtypes have been characterized ($P2Y_1$, $P2Y_2$, $P2Y_4$, $P2Y_6$, $P2Y_{11-14}$). Recently, several studies have shed light on the role of P2Y receptors in bone biology. Among them, little is known on the role of $P2Y_6$ receptor on osteoclast differentiation. Thus, we investigated the role of $P2Y_6$ receptor on osteoclastogenesis using $P2Y_6$ receptor selective antagonist, MRS 2578. When osteoblasts and bone marrow cells were co-cultured in the presence of $VitD_3$ and $PGE_2$, $P2Y_6$ antagonist increased the formation of TRAP positive osteoclasts. To elucidate the target cells of $P2Y_6$ antagonist, we first checked the effect of MRS 2578 on osteoblasts. Treatment of MRS 2578 did not affect OPG : RANKL mRNA ratio in osteoblasts. Next, we checked the effects of $P2Y_6$ antagonist on osteoclast precursors using mouse bone marrow macrophages (BMMs). Addition of MRS 2578 increased the number of osteoclasts in RANKL-treated BMMs. Although $P2Y_6$ antagonist had no effect on RANKL-induced NFATc1, c-Fos and MafB expression levels, it significantly stimulated RANKL-induced Blimp1 mRNA expression in BMMs. Taken together, these data indicate that $P2Y_6$ antagonist increases osteoclast formation by upregulation of Blimp1 expression.

Evaluation for detection of Cryptosporidium oocysts in diarrheal feces of calves (야외 송아지 설사변에서 작은와포자충 검출에 대한 평가)

  • Wi, Seong-Hwan;Ju, Hu-Don;Gang, Yeong-Bae
    • Parasites, Hosts and Diseases
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    • v.34 no.2
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    • pp.121-126
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    • 1996
  • For the detection of Cwptospori,mum oocysts, fecal samples were collected from 201 calves which showed diarrhea. Among the 201 samples, 29 samples (14.4%) were positive for Cwptosporinium spry. by the DMSO-modified acid-fast stain (MAFS) , 23 samples (11.4%) were positive by commercial kit (Meridian Diagnostics, Cincinnati, Ohiol and 23 by the indirect immunofluorescence antibody (IFA )assay employing the monoclonal antibody (mAb C6). When tested by both IFA and MAFS, 20 fecal samples were positive for Cwptosporinium oocysts whereas 169 fecal samples were negative. If the MAFS is considered a standard method for oocyst detection, the IFA showed 69% of sensitivity and 98% of specificity. When tested by both IFA and commercial kit, 22 fecal samples were positive for Cwptospori,mum oocysts while 177 samples were negative. One sample tested by IFA was found to be false negative, when compared with the results by commercial kit. The sensitivity of IFA was calculated as high as 96%; the specificity as 99% and the predictive value was also 99%. In the present study, IFA employing the nAb C6 revealed that 23 samples (11.4%) were positive among the 201 calves showing diarrhea. Of 23 IFA positive samples, 4 samples (5%) showed cryptosporidial oocysts more than 105 OPG Therefore. it is concluded that the calves showing cryptosporidial oocysts more than 105 OPG in the feces were highly associated with clinical cryptosporidiosis.

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EFFECT OF CALCIUM HYDROXIDE REMNANT AS AN INTRACANAL DRESSING ON THE APICAL LEAKAGE (근관내 잔존 Calcium Hydroxide제재가 치근단 미세누출에 미치는 영향)

  • 신수일;조용범
    • Restorative Dentistry and Endodontics
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    • v.26 no.1
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    • pp.77-85
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    • 2001
  • The purpose of this in vitro study was to compare the apical leakage in extracted teeth filled with gutta-percha subsequent to dressing with one of three different calcium hydroxide preparations. Thirty six extracted teeth with single canal were used in this study. After working length determination, canals were prepared with K flexo files to a #40 at the working length. Step-back flaring was produced by using #45, #50 K flexo files and #2, #3, #4 Gates Glidden burs. The teeth were randomly divided into 3 groups of 10 each : the remaining six teeth were used for negative and positive leakage control: Group 1, dressed with pure calcium hydroxide powder (Sigma, USA) mixed with distilled water; Group 2, dressed with Metapaste (Metadent, Korea) ; Group 3, dressed with Vitapex (Neo Dental, Japan). Teeth were sealed with Caviton (GC, Japan) and incubated in 100% humidity, at 37$^{\circ}C$ for 1 wk. All kinds of calcium hydroxide were removed from the canal with a MAF and 5% NaOCl. The canals were filled with AH-26$^{\circledR}$ sealer and gutta-percha using lateral condensation technique, incubated in 100% humidity, at 37$^{\circ}C$ for 2 days for the sealer to be set. The teeth were coated twice with nail varnish except for an area of approximately 2mm surrounding the apical foramen. All specimens were placed in 2% methylene blue solution for 2 days. The root were sectioned longitudinally, the amount of apical leakage was measured to the most coronal part of the root canal to which the dye had penetrated. The independent measurements were made for each root using a stereomicroscope ($\times$40 magnification) and the average was recorded for statistical analysis. The results were as follows ; 1. The mean of apical leakage in group of pure calcium hydroxide ranged 0.102$\pm$0.156mm, in Metapaste$^{\circledR}$ ranged 0.062$\pm$0.069mm, and in Vitapex$^{\circledR}$ ranged 0.067$\pm$0.072mm. 2. Group of pure calcium hydroxide exhibited more leakage than those of 2 manufactured calcium hydroxide preparations, but it was not statistically significant. 3. Group of water-based Metapaste$^{\circledR}$ showed lesser leakage than that of oil-based Vitapex$^{\circledR}$, but it was not statistically significant.

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Association Study in Endothelin 1 (EDN1) Gene Polymorphism and Excess or Deficiency Syndrome in Korean Asthmatic Patients (한국인 기관지 천식 허증(虛證), 실증(實證) 환자와 EDN1 유전자 다형성과의 상관성 연구)

  • Yoem, Yu-rim;Kim, Kwan-il;Baek, Hyun-jung;Kim, Mi-a;Lee, Beom-joon;Kim, Jin-ju;Kim, Su-kang;Chung, Joo-ho;Jung, Hee-jae;Jung, Sung-ki
    • The Journal of Internal Korean Medicine
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    • v.37 no.1
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    • pp.47-64
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    • 2016
  • Objectives: In the present study, a genetic analysis was conducted to investigate the association of the expression of SNPs of EDN1 gene polymorphism with the clinical phenotype in bronchial asthma patients with either excess or deficiency syndrome.Methods: Ninety-four healthy control subjects and 52 asthma patients were included in this study. The asthma patients were divided into two groups: those with deficiency syndrome and those with excess syndrome. We searched the exonic and promoter areas of the EDN1 gene in the NCBI website SNPs with <0.01 minor allele frequency (MAF) and <0.01 heterozygosity. Pro programs were performed to obtain the odds ratio, 95% confidence interval, and p-value. Multiple logistic regression models were conducted to analyze the genetic data.Results: In our genotype and allele analyses, there were significant differences in the codominant 2 model of the rs3087459 SNP genotype and also in the CGG haplotype between the control group and the asthma group. Genotype and allele analyses were conducted between the deficiency and excess syndrome group. There were significant differences in the dominant and log-additive model and also in the frequency of C-alleles of rs3087459 SNP genotype. There were significant differences in codominant 1, dominant and log-additive model and T-allele of rs5370 SNP genotype. The AGG haplotype also revealed significant differences.Conclusions: EDN1 SNPs (rs3087459, rs5370) showed a significant association with symptomatic excess syndrome in Korean asthmatic patients.

Exonic SNP (rs7144, 3’-UTR) in CD46 Molecule and Complement Regulatory Protein (CD46) Gene Associated with Excess Syndrome to Categorize Korean Bronchial Asthma Patients (한국인 기관지 천식 허증(虛證), 실증(實證) 환자와 CD46 유전자 다형성과의 관계)

  • Lee, Mei;Baek, Hyun-jung;Park, Eui-keun;Kim, Kwan-il;Lee, Beom-joon;Kim, Su-kang;Chung, Joo-ho;Kim, Jin-ju;Kim, Mi-a;Jung, Hee-jae;Jung, Sung-ki
    • The Journal of Internal Korean Medicine
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    • v.36 no.4
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    • pp.547-561
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    • 2015
  • Objectives In this study, we divided Korean asthma patients into excess syndrome or deficiency syndrome groups according to clinical phenotype. Genetic analysis was conducted to investigate the association of exonic SNPs in the CD46 gene polymorphism with the clinical phenotype based on the differentiation syndrome of the bronchial asthma patients.Methods There were 95 healthy patients (control group) and 53 asthma patients. (The deficiency syndrome group included 24 and the excess syndrome group 29). We searched the exonic areas of the CD46 gene in the NCBI website SNPs with <0.01 minor allele frequency (MAF) and <0.01 heterozygosity. We finally selected two SNPs: rs138843816, Ser13Phe and rs7144, 3’-UTR. Hardy-Weinberg equilibrium was calculated using SNPStats.Results There were significant differences in the codominant 1 model and the dominant model between the healthy group and the asthma group. There were significant differences between deficiency syndrome group and the excess syndrome group in the genotype frequencies and in the codominant 1 model, the dominant model, and the log-additive model. The allele frequency of rs7144C showed a significant difference between the deficiency syndrome group and the excess syndrome group. Two-SNP haplotype analysis showed a significant difference in frequency in the deficiency syndrome group and in the excess syndrome group. There were significant differences between the healthy group and the excess syndrome group in the codominant 1 model, the dominant model, and the log-additive model. The frequency of the rs7144 C allele exhibited a significant difference in the demonstration. SNP haplotype analysis between the healthy group and the excess syndrome group showed a significant difference in the frequency of the CT haplotype and the CC haplotype.Conclusions The results indicate that two CD46 SNPs (rs138843816, Ser13Phe and rs7144, 3′–UTR) might be associated with the symptomatic excess syndrome in Korean asthma patients.