• The Korea Journal of Herbology
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• v.21 no.2
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• pp.37-46
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• 2006

Objectives : The purpose of this study was to investigate the effect of Dangguibohyultang (DB) and its combination (DB-I; Astragali membraneus BUNGE : Angelica gigas NAKAI=5:1, DB-II; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:1, DB-III; Astragali membraneus BUNGE:Angelica gigas NAKAI=1:5,) on apoptosis in human colorectal adenocarcinoma HCT116 cells. Methods : To study the cytotoxic effect of methanol extract of DB-I, DB-II and DB-III on HCT116 cells, the cell viability was determined by XTT reduction method and ttypan blue exclusion assay. To confirm the induction of apoptosis, the cleavage of poly ADP-ribose polymerase (PARP), a substrate for caspase-3 and a typical sign of apoptosis, and the activation of procaspase-3, -8 and -9 were examined by western blot analysis. Furthermore, DB-induced apoptosis was confirmed by DNA fragmentation. The release of cytochrome C from mitochondria to cytosol, the level of Bcl-2 and Bax, and the expressions of Raf/MEK/ERK were examined by western blot analysis. Results : DB-I and DB-II reduced proliferation of HCT116 cells in a dose-dependent manner. DB-I and DB-II decreased procaspase-3, -8, -9 levels in a dose-dependent manner and induced the clevage of PARP. DB-I and DB-II also triggered the mitochondrial apoptotic signaling by increasing the release of cytochrome C from mitochondria to cytosol, decreasing of anti-apoptotic Bcl-2, and increasing of pro-apoptotic Bax. DB-I and DB-II decreased the activation of Ras/Raf/MEK/ERK cascade in a dose-dependent manner. Conclusion : These results suggest that DB-I and DB-II induce apoptosis via mitochondrial pathway in HCT116 cells. Furthermore, Raf/MEK/ERK cascade is involved in DB-induced apoptosis. These results suggest that DB is potentially useful as a chemotherapeutic agent in human liver cancer.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.6
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• pp.483-490
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• 2001

There were many controversies in the cause and progress of tumorigenesis. Recently, studies on the mutation of genes related to the tumor have extensively been performed due to development of molecular biology. Structural and morphological changes of chromosomes, which are related to the abnormal activation of oncogenes or inactivation of tumor suppression genes, transform the normal cells into the tumor cells. p53 and Rb are well known tumor suppressor genes, while oncogenes include c-myc, bcl-2 and ras, etc. When exposed to cell damaging agents, p53 inhibits cell growth by inducing transcription of p21. Especially p73, which is homo-logy of p53, frequently deleted in melanoma, neuroblastoma, colon cancer, and breast cancer, when over produced, p73 activates the transcription of p21, bax-1 and inhibits cell growth by inducing apoptosis. For study on mRNA expression of p21 and p73, normal oral keratinocytes, and cell lines of primary and metastatic oral squamous cell carcinoma were cultured and then electrophoresis and RT-PCR(reverse transcription-polymerase chain reaction) were performed. 1. The mRNA of p21 and p73 in normal oral keratinocyte expressed lower than that of primary squamous cell carcinoma. 2. The mRNA of p21 in metastatic oral squamous carcinoma cell lines was expressed as various patterns compared with that of normal oral keratinocyte. 3. In the metastatic oral squamous cell lines, the mRNA of HN8 expressed higher than that of HN12 or HN19. 4. The mRNA of p73 in primary oral squamous cell lines expressed 4-5 times higher than that of normal keratinocyte. 5. In metastatic oral squamous cell lines, there was no significant expression of p73 mRNA compared with that of normal oral keratinocyte. From the results obtained in this study, mRNA expression of p73 in primary oral squamous cell lines was remarkable, while mRNA expression of p21 and p73 in metastatic oral squamous cell lines were statistically insignificant.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.1
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• pp.201-208
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• 2013

Objective: To investigate changes in the invasive capacity of gastric cancer cells in vitro after expression inhibition of T lymphoma invasion and metastasis inducing factor 1 (Tiam 1) and underlying mechanisms. Methods: Using adhesion selection, two subpopulations with high ($M_H$) or low ($M_L$) invasive capacity were separated from the human gastric cancer cell line MKN-45 ($M_0$). Tiam 1 antisense oligodeoxynucleotide (ASODN) was transfected into $M_H$ cells with liposomes, and expression of Tiam 1 mRNA and protein was determined by RT-PCR and quantitative cellular-ELISA. Changes in the cytoskeleton, invasive capacity in vitro and expression of ras-related $C_3$ botulinum toxin substrate 1 (Rac 1), integrin ${\beta}1$ and matrix metalloproteinase 2 (MMP 2) between Tiam 1 ASODN transfected $M_H$ cells and non-transfected cells were observed by HE staining, cytoskeletal protein staining, scanning electron microscopy, Boyden chamber tests and cyto-immunohistochemistry. Results: A positive correlation existed between the expression level of Tiam l mRNA or protein and the invasion capacity of gastric cancer cells. After ASODN treatment ($0.43{\mu}M$ for 48 h), Tiam 1 mRNA transcription and protein expression in $M_H$ cells were decreased by 80% and 24% respectively (P < 0.05), compared with untreated controls, while invasive capacity in vitro was suppressed by 60% (P < 0.05). Morphologic and ultrastructural observation also showed that ASODN-treated $M_H$ cells exhibited smooth surfaces with obviously reduced filopodia and microspikes, which resembled $M_0$ and $M_L$ cells. Additionally, cytoskeletal distribution dramatically altered from disorder to regularity with reduced long filament-like structure, projections, pseudopodia on cell surface, and with decreased acitn-bodies in cytoplasm. After Tiam 1 ASODN treatment, the expression of Rac 1 and Integrin ${\beta}1$ in $M_H$ cells was not affected (P > 0.05), but that of MMP 2 in $M_H$ cells was significantly inhibited compared with untreated cells (P < 0.05). Conclusion: Over-expression of Tiam-1 contributes to the invasive phenotype of gastric cancer cells. Inhibition of Tiam 1 expression could impair the invasive capacity of gastric cancer cells through modulating reconstruction of the cytoskeleton and regulating expression of MMP 2.

• Molecules and Cells
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• v.26 no.6
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• pp.566-575
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• 2008

Twenty-five chipmunk species occur in the world, of which only the Siberian chipmunk, Tamias sibiricus, inhabits Asia. To investigate mitochondrial cytochrome b sequence variations and population structure of the Siberian chipmunk in northeastern Asia, we examined mitochondrial cytochrome b sequences (1140 bp) from 3 countries. Analyses of 41 individuals from South Korea and 33 individuals from Russia and northeast China resulted in 37 haplotypes and 27 haplotypes, respectively. There were no shared haplotypes between South Korea and Russia - northeast China. Phylogenetic trees and network analysis showed 2 major maternal lineages for haplotypes, referred to as the S and R lineages. Haplotype grouping in each cluster was nearly coincident with its geographic affinity. In particular, 3 distinct groups were found that mostly clustered in the northern, central and southern parts of South Korea. Nucleotide diversity of the S lineage was twice that of lineage R. The divergence between S and R lineages was estimated to be 2.98-0.98 Myr. During the ice age, there may have been at least 2 refuges in South Korea and Russia - northeast China. The sequence variation between the S and R lineages was 11.3% (K2P), which is indicative of specific recognition in rodents. These results suggest that T. sibiricus from South Korea could be considered a separate species. However, additional information, such as details of distribution, nuclear genes data or morphology, is required to strengthen this hypothesis.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6155-6157
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• 2012

Background: Mutations in the MAPK (Mitogen Activated Protein Kinase) signaling pathway - EGFR/Ras/RAF/MEK have been associated with the development of several carcinomas. ERK2, a downstream target of the MAPK pathway and a founding member of the MAPK family is activated by cellular signals emanating at the cell membrane. Activated ERK2 translocates into the nucleus to transactivate genes that promote cell proliferation. MKP - a dual specific phosphatase - interacts with activated ERK2 via the common docking (CD) domain of the later to inactivate (dephosphorylate) and effectively terminate further cell proliferation. A constitutively active form of ERK2 carrying a single point mutation - E322K in its CD domain, was earlier reported by our laboratory. In the present study, we investigated the prevalence of this CD domain E322K mutation in 88 well differentiated OSCC tissue samples. Materials and Method: Genomic DNA specimens isolated from 88 oral squamous cell carcinoma tissue samples were amplified with primers flanking the CD domain of the ERK2 gene. Subsequently, PCR amplicons were gel purified and subjected to direct sequencing to screen for mutations. Results: Direct sequencing of eighty eight OSCC samples identified an E322K CD domain mutation in only one (1.1%) OSCC sample. Conclusions: Our result indicates that mutation in the CD domain of ERK2 is rare in OSCC patients, which suggests the role of genetic alterations in other mitogenic genes in the development of carcinoma in the rest of the patients. Nevertheless, the finding is clinically significant, as the relatively rare prevalence of the E322K mutation in OSCC suggests that ERK2, being a common end point signal in the multi-hierarchical mitogen activated signaling pathway may be explored as a viable drug target in the treatment of OSCC.

• Korean Journal of Ichthyology
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• v.29 no.1
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• pp.13-21
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• 2017

We investigated that changes of survival rate and hematological factors in sevenband grouper (Epinephelus septemfasciatus), exposed to ammonia. First, the fish was exposed and was observed survival rate. We observed the survival rate of the fish when exposed at each ammonia concentrations (0, 0.35, 0.4, 0.5, 0.8 mg/L, $NH_3$) for 96 hours. The 96-h $LC_{50}$ (lethal ambient concentration for 50% of the population) of $NH_4^+$ and un-ionized ammonia ($NH_3$) for sevenband grouper were 26.7 and 0.6 mg/L, respectively. Secondly, we exposed the fish to 96-h $LC_{50}$ ammonia concentration (0.6 mg/L) for 96 hours, then sampled the blood of groupers for analysis of cortisol, glucose, $NH_3$, GOT, GPT, total protein (TP), electrolytes ($Na^+$, $Cl^-$) and osmolality in plasma. The plasma cortisol, glucose, GOT and GPT were increased with increasing of elapsed time and ammonia concentration. In addition, the $NH_3$ and total protein in plasma were significantly increased to 12 hours exposed to ammonia, and then decreased. But, the values were significantly higher than the control. The plasma electrolytes ($Na^+$, $Cl^-$) and osmolality were decreased with increasing of elasped time. The results in the present study suggest that the continuous exposure of ammonia to juveniles of sevenband grouper may be a stressor and negative influence.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.11
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• pp.1529-1539
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• 2017

Objective: The objective of this study was to compare the DNA methylation profile in the longissimus dorsi muscle between Small Tailed Han and Dorper${\times}$Small Tailed Han crossbred sheep which were known to exhibit significant difference in meat-production. Methods: Six samples (three in each group) were subjected to the methylated DNA immunoprecipitation sequencing (MeDIP-seq) and subsequent bioinformatics analyses to detect differentially methylated regions (DMRs) between the two groups. Results: 23.08 Gb clean data from six samples were generated and 808 DMRs were identified in gene body or their neighboring up/downstream regions. Compared with Small Tailed Han sheep, we observed a tendency toward a global loss of DNA methylation in these DMRs in the crossbred group. Gene ontology enrichment analysis found several gene sets which were hypomethylated in gene-body region, including nucleoside binding, motor activity, phospholipid binding and cell junction. Numerous genes were found to be differentially methylated between the two groups with several genes significantly differentially methylated, including transforming growth factor beta 3 (TGFB3), acyl-CoA synthetase long chain family member 1 (ACSL1), ryanodine receptor 1 (RYR1), acyl-CoA oxidase 2 (ACOX2), peroxisome proliferator activated receptor-gamma2 (PPARG2), netrin 1 (NTN1), ras and rab interactor 2 (RIN2), microtubule associated protein RP/EB family member 1 (MAPRE1), ADAM metallopeptidase with thrombospondin type 1 motif 2 (ADAMTS2), myomesin 1 (MYOM1), zinc finger, DHHC type containing 13 (ZDHHC13), and SH3 and PX domains 2B (SH3PXD2B). The real-time quantitative polymerase chain reaction validation showed that the 12 genes are differentially expressed between the two groups. Conclusion: In the current study, a tendency to a global loss of DNA methylation in these DMRs in the crossbred group was found. Twelve genes, TGFB3, ACSL1, RYR1, ACOX2, PPARG2, NTN1, RIN2, MAPRE1, ADAMTS2, MYOM1, ZDHHC13, and SH3PXD2B, were found to be differentially methylated between the two groups by gene ontology enrichment analysis. There are differences in the expression of 12 genes, of which ACSL1, RIN2, and ADAMTS2 have a negative correlation with methylation levels and the data suggest that DNA methylation levels in DMRs of the 3 genes may have an influence on the expression. These results will serve as a valuable resource for DNA methylation investigations on screening candidate genes which might be related to meat production in sheep.

• Journal of Korea Water Resources Association
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• v.50 no.10
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• pp.661-671
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• 2017

The objective of this study was to analyze the flood stage considering the uncertainty caused by the river roughness coefficients and discharge. The methodology of this study involved the GLUE (Generalized Likelihood Uncertainty Estimation) to quantify the uncertainty bounds applying three different storm events. The uncertainty range of the roughness was 0.025~0.040. In case of discharge, the uncertainty stemmed from parameters in stage-discharge rating curve, if h represents stage for discharge Q, which can be written as $Q=A(h-B)^C$. Parameters in rating curve (A, B and C) were estimated by non-linear regression model and assumed by t distribution. The range of parameters in rating curve was 5.138~18.442 for A, -0.524~0.104 for B and 2.427~2.924 for C. By sampling 10,000 parameter sets, Monte Carlo simulations were performed. The simulated stage value was represented by 95% confidence interval. In storm event 1~3, the average bound was 0.39 m, 0.83 m and 0.96 m, respectively. The peak bound was 0.52 m, 1.36 m and 1.75 m, respectively. The recurrence year of each storm event applying the frequency analysis was 1-year, 10-year and 25-year, respectively.

• Journal of Wetlands Research
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• v.13 no.1
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• pp.149-160
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• 2011

This study developed a HSI for the creatures in Hongcheon River in order to analyze the conditions proper for habitats. For the index, the investigator identified a total of seven items encompassing hydraulic characteristics such as flow velocity and water depth, and water quality characteristics such as water temperature, BOD, DO, TN, and TP. The subject river was simulated, inspected, and revised with a two-dimensional river model (RMA-2) and water quality model (QUAL2E). Using GIS, the developed index was divided by section by reflecting river characteristics and compared and analyzed with the statistics. The river was divided into a total of 29 reaches by reflecting the basic characteristics and the features of the hydraulic coefficient on the cross-sections of the river. According to the analysis results, the fish scored the highest mean of the overall habitat suitability index of 0.769 at reach 27. Each of the variables had the following mean values: 0.122 m/s for flow velocity, 0.782m for water depth, $14.3^{\circ}C$ for water temperature, 0.68 mg/l for BOD, 10.3 mg/l for DO, 2.4 mg/l for TN, and 0.0121mg/l for TP.

• Journal of Korea Water Resources Association
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• v.48 no.1
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• pp.45-55
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• 2015

In this study, when seawall or harbor gate is installed for coastal disaster prevention, a two-dimensional water analysis in the bay is carried out to consider the flood amount of river inflow and effect of harbor gate. The Yeongsan river and the port Mokpo area are selcected for the study region. Then, by analyzing the hydraulic characteristics of flood flow of the Yeongsan river, we analysed the compatibility of the results in the two-dimensional hydrodynamic model. A tw-odimensional water analysis were conducted for the four cases considering whether a harbor gate is installed or not, and whether the inland water boundary condition is considered or not, also with open sea boundary condition. The results of the two-dimensional water analysis shows that water level change near the port Mokpo area is mainly caused by the discharge of the estuary barrage of the Yeongsan river because the harbor gate was installed. In addition, it is revealed that the volume of reservoir created by the harbor gate and the estuary barrage is too much small compared to the volume of the discharge from the Yeongsan river. Therefore, when the harbor gate is installed in the open sea, we concluded that a flexible management between the harbor gate and the estuary barrage of the Yeongsan river is required. A initial water level of the bay and outflow from the harbor gate are proposed for disaster prevention in the coastal area of port Mokpo.