• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.245-252
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• 2014

Human METCAM/MUC18, a cell adhesion molecule (CAM) in the immunoglobulin-like gene super family, plays a dual role in the progression of several epithelium cancers; however, its role in the nasopharyngeal carcinoma (NPC) remains unclear. To initiate the study we determined human METCAM/MUC18 expression in tissue samples of normal nasopharynx (NP), NPCs, and metastatic lesions, and in two established NPC cell lines. Immunoblotting analysis was used for the determination in lysates of frozen tissues, and immunohistochemistry (IHC) for expression in formalin-fixed, paraffin-embedded tissue sections of 7 normal nasopharynx specimens, 94 NPC tissue specimens, and 3 metastatic lesions. Human METCAM/MUC18 was expressed in 100% of the normal NP, not expressed in 73% of NPC specimens (or expressed at very low levels in only about 27% of NPC specimens), and expressed again in all of the metastatic lesions. The level of human METCAM/MUC18 expression in NPC tissues was about one fifth of that in the normal NP and metastatic lesions. The low level of human METCAM/MUC18 expression in NPC specimens was confirmed by a weak signal of RT-PCR amplification of the mRNA. Low expression levels of human METCAM/MUC18 in NPC tissues were also reflected in the seven established NPC cell lines. These findings provided the first evidence that diminished expression of human METCAM/MUC18 is an indicator for the emergence of NPC, but increased expression then occurs with metastatic progression, suggesting that huMETCAM/MUC18, perhaps similar to TGF-${\beta}$, may be a tumor suppressor, but a metastasis promoter for NPC.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.22
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• pp.9791-9795
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• 2014

To study the gene expression change and possible signal pathway during androgen-dependent prostate cancer (ADPC) becoming androgen-independent prostate cancer (AIPC), an LNCaP cell model of AIPC was established using flutamide in combination with androgen-free environment inducement, and differential expression genes were screened by microarray. Then the biological process, molecular function and KEGG pathway of differential expression genes are analyzed by Molecule Annotation System (MAS). By comparison of 12,207 expression genes, 347 expression genes were acquired, of which 156 were up-ragulated and 191 down-regulated. After analyzing the biological process and molecule function of differential expression genes, these genes are found to play crucial roles in cell proliferation, differntiation, cell cycle control, protein metabolism and modification and other biological process, serve as signal molecules, enzymes, peptide hormones, cytokines, cytoskeletal proteins and adhesion molecules. The analysis of KEGG show that the relevant genes of AIPC transformation participate in glutathione metabolism, cell cycle, P53 signal pathway, cytochrome P450 metabolism, Hedgehog signal pathway, MAPK signal pathway, adipocytokines signal pathway, PPAR signal pathway, TGF-${\beta}$ signal pathway and JAK-STAT signal pathway. In conclusion, during the process of ADPC becoming AIPC, it is not only one specific gene or pathway, but multiple genes and pathways that change. The findings above lay the foundation for study of AIPC mechanism and development of AIPC targeting drugs.

• Journal of Applied Biological Chemistry
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• v.50 no.2
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• pp.78-84
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• 2007

The anti-fibrotic effects of hot water extract of adventitious root culture of Panax ginseng (ARCP) and the possible mechanisms were investigated on $CCl_4-induced$ hepatotoxicity model mice. Fibrosis was induced by a mild treatment of $CCl_4$. Then silymarin as a positive control drug and ARCP or carrier alone as a negative control were treated. Serum GPT, GOT and ALP activity levels were lowered by 25, 21 and 11% for silymarin treated group and by 48, 39 and 14% for ARCP treated group compared to carrier treated alone. Hepatic collagen for ARCP treatment group was reduced by 18% and MDA contents decreased a little more. Pro-fibrotic gene ($TGF-{\beta}1$, TIMP-1 and ${\alpha}-SMA$) expression increased following the $CCl_4$ treatment, but both the silymarin and the ARCP treatments decreased the expressions of these genes by 20% to 50%. The antioxidant effect of ARCP was studied by DPPH free radical scavenging activity. In addition, a generation of reactive oxygen species (ROS) was also reduced in $H_2O_2-treated$ HepG2 cells upon the ARCP treatment. In summary, ARCP has antioxidant property, and can have some protection against oxidative stress; more importantly, ARCP can efficiently protect mice against $CCl_4-induced$ fibrosis.

• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2000.04a
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• pp.178-181
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• 2000

In order to develop the novel gene expression system, we introduced new control elements which could influence the foreign gene expression in animal cells. When the foreign genes are introduced into the genome of higher eukaryotic cells, the expressions from these integrated genes are often low and can vary greatly depending on the positions of the integration sites due to the complex nature of the chromatin structures (1). First we screened the various DNA sequence elements which can function as an insulator of gene expression from these position effects and can cooperate with the SV40 enhancer/promoter. Among the several DNA elements from the various sources, we identified the particular DNA element which confers the increased frequency of the positive colonies, assayed by the reporter gene from stable selections indicating significantly reduced position effects. This element also showed the several fold-increased expression level as well as the copy-number dependent expression with host cell specificity. Second we modified the transcription termination element where we introduced the specific terminator in combination with SV40 polyA signal. This modified terminator showed the increased efficiency and the level of the gene expression. By combining these two elements, we made the animal cell expression system and tested successfully for the recombinant protein productions of TGF ${\beta}$-soluble receptor, Antithrombin III, and single chain Pro-Urokinase. [Supported by grants from MOCIE]

• The korean journal of orthodontics
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• v.42 no.4
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• pp.207-217
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• 2012

Objective: This study aimed to evaluate the effect of an intentionally created socket on bone remodeling with orthodontic tooth movement in rabbits. Methods: Eighteen male rabbits weighing 3.8 - 4.25 kg were used. An 8-mm deep and 2-mm wide socket was drilled in the bone 1 mm mesial to the right mandibular first premolar. The left first premolar was extracted to serve as an extraction socket. A traction force of 100 cN was applied to the right first premolar and left second premolar. Sections were obtained at the middle third of the moving tooth for both the drilled and extraction sockets and evaluated with hematoxylin and eosin staining and immunohistochemical analyses. The amount of tooth movement and tartrate-resistant acid phosphatase (TRAP)-positive cell count were compared between the 2 groups using the Mann-Whitney U test. Results: At week 2, the distance of tooth movement was significantly higher in the intentional socket group (p < 0.05) than in the extraction socket group. The number of TRAP-positive cells decreased in week 2 but increased in week 3 (p < 0.05). However, there were no significant differences between the groups. Furthermore, results of transforming growth factor (TGF)-${\beta}$ staining revealed no significant differences. Conclusions: The intentional socket group showed greater distance of tooth movement than did the extraction socket group at week 2. Osteoclast counts and results of immunohistochemical analyses suggested elevated bone remodeling in both the groups. Thus, osteotomy may be an effective modality for enhancing tooth movement in orthodontic treatment.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.10
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• pp.4217-4224
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• 2014

Cholangiocarcinoma (CCA), a slow growing but highly metastatic tumor, is highly prevalent in Northeast Thailand. Specific tests that predict prognosis of CCA remain elusive. The present study was designed to investigate whether peripheral blood leukocyte (PBL) transcriptional profiles might be of use as a prognostic test in CCA patients. Gene expression profiles of PBLs from 9 CCA and 8 healthy subjects were conducted using the Affymetrix HG_U133 Plus 2.0 GeneChip. We indentified informative PBLs gene expression profiles that could reliably distinguish CCA patients from healthy subjects. Of these CCA specific genes, 117 genes were up regulated and 60 were down regulated. The molecular and cellular functions predicted for these CCA specific genes according to the Gene Ontology database indicated differential PBL expression of host immune response and tumor progression genes (EREG, TGF ${\beta}1$, CXCL2, CXCL3, IL-8, and VEGFA). The expression levels of 9 differentially expressed genes were verified in 36 CCA vs 20 healthy subjects. A set of three tumor invasion related genes (PLAU, CTSL and SERPINB2) computed as "prognostic index" was found to be an independent and statistically significant predictor for CCA patient survival. The present study shows that CCA PBLs may serve as disease predictive clinically accessible surrogates for indentifying expressed genes reflective of CCA disease severity.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.3
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• pp.227-237
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• 2008

STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

• IMMUNE NETWORK
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• v.12 no.6
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• pp.269-276
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• 2012

The anti-tumor effect of monocyte-derived DC (MoDC) vaccine was studied in lung cancer model with feasible but weak Ag-specific immune response and incomplete blocking of tumor growth. To overcome this limitation, the hematopoietic stem cell-derived DC (SDC) was cultured and the anti-tumor effect of MoDC & SDC was compared in mouse lung cancer minimal residual model (MRD). Therapeutic DCs were cultured from either $CD34^+$ hematopoietic stem cells with GM-CSF, SCF and IL-4 for 14 days (SDC) or monocytes with GM-CSF and IL-4 for 7 days (MoDC). DCs were injected twice by one week interval into the peritoneum of mice that are inoculated with Lewis Lung Carcinoma cells (LLC) one day before the DC injection. Anti-tumor responses and the immune modulation were observed 3 weeks after the final DC injection. CD11c expression, IL-12 and TGF-${\beta}$ secretion were higher in SDC but CCR7 expression, IFN-${\gamma}$ and IL-10 secretion were higher in MoDC. The proportion of $CD11c^+CD8a^+$ cells was similar in both DC cultures. Although both DC reduced the tumor burden, histological anti-tumor effect and the frequencies of IFN-${\gamma}$ secreting $CD8^+$ T cells were higher in SDC treated group than in MoDC. Conclusively, although both MoDC and SDC can induce the anti-tumor immunity, SDC may be better module as anti-tumor vaccine than MoDC in mouse lung cancer.

• International Journal of Oral Biology
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• v.33 no.2
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• pp.51-58
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• 2008

A mutation of UNCL, an inner nuclear membrane RNAbinding protein, has been found to eliminate mechanotransduction in Drosophila. UNCL is expressed in human periodontal tissue including in periodontal ligament (PDL) fibroblasts. However, it is unclear how a mechanical stimulus is translated into cellular responses in PDL fibroblasts. The aim of this study was to evaluate the effect of UNCl on mechanical stress related genes in PDL fibroblasts in response to mechanical stress. The mRNA of TGF-$\beta$, COX-2, and MMP-2 was up-regulated after UNCL inactivation in PDL fibroblasts under the compression force. Under the tensile force, inactivation of UNCL decreased the expression of Biglycan, RANKL, MMP-2, and TIMP-2 mRNAs while it increased the expression of TIMP-1. p38-MAPK was expressed in PDL fibroblasts under compression forces whereas phospho-ERK1/2, p65-NFkB, and c-fos were expressed under tension forces. The expression and phosphorylation of the mechanical stress related genes, kinases, and transcription factors were changed according to the types of stress. Furthermore, most of them were regulated by the inactivation of UNCL. This suggests that UNCL is involved in the regulation of mechanical stress related genes through the signaling pathway in PDL fibroblasts.

• Korean Journal of Oriental Medicine
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• v.8 no.2 s.9
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• pp.95-107
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• 2002

Yukmijihwang-tang(YM) is a noted herbal prescription in Chinese and Korean traditional medicines, and it has been known to reinforce the vital essence and has been widely used for a variety of disease such as stroke, osteoporosis, anti-tumor, and hypothyrodism. Regarding its traditional use, YM has been known to reinforce the Yin (vital essence) of liver and kidney. Also it has been known to reinforce nutrition and biological function in brain. Recently, studies suggested that YM increase antioxidant activities and exert the protective effect against oxidant-induced liver cell injury. We investigated the high-throughput gene expression analysis on the Yukmijihwang-tang administrated in SD rats. Microarray data were validated on a limited number of genes by semiquantitative RT-PCR and Western blot analyses. The recent availability of microarrays provides an attractive strategy for elaborating an unbiased molecular profile of large number of genes in drug discovery This experimental approach offers the potential to identify molecules or cellular pathways not previously associated with herbal medicine. Total RNA from normal control brain and Yukmijihwang-tang administrated brain were hybridized to microarrays containing 10,000 rat genes. The 52 genes were found to be up-regulated(twice or more) excluding EST gene. The nine genes were found to be down-regulated(twice or more) excluding EST gene. Gene array technology was used to identify for the first time many genes expression pathway analysis that arecell cycle pathway, apoptosis pathway, electron transport chain pathway, cytoplasmic ribosomal protein pathway, fatty acid degradation pathway, and TGF-beta signaling pathway. These differentially expressed genes pathway analysis have not previously been iavestigated in the context of herbal medicine efficacy and represent novel factors for further study of the mechanism of herbal medicine efficacy.