• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.291-310
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• 1998

This study was performed to evaluate the effects of extracts of Drynariae Rhizoma on the characteristics of rat calvaria cells(RCV) and bone marrow cells(RBM) which have the important role on the bone formation in vitro. Drynariae Rhizoma has been known as the useful herbal medicament for treatment of the wound healing including regeneration of bone fracture, and also has been used to treat the periodontal lesions, tooth mobility, gingival bleeding and pus discharge via sulcus in Oriental Medicine. In control group, the cells were cultured alone with Dulbeco's Modified Eagle's Medium contained with 10% fetal bovine serum, 100U/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, extracts of Drynariae Rhizoma(0.1, 1, 5, 10, $50{\mu}g/ml$) were added into the above culture condition. And then each group was characterized by examing the cell proliferation at 1, 3, 7, 14, 21, 30th day, the amount of total protein synthesis and alkaline phosphatase activity of RCV at 2,4th day and those of RBM at 3, 6th day. And also, the calcified nodule of RCV was examed at 3, 5th day in three goup, control, experimental, culture with the PDGF group. The results were as follow ; 1. Both RCV and RBM cells in Drynariae Rhizoma-treated experimental group proliferated more rapidly than nontreated control group. The experimental group below $5{\mu}g/ml$ Drynariae Rhizoma-treated showed more prominent cell proliferation from the 7th day to the 21st day than the control group and above $10\;{\mu}g/ml$ treated group in RCV. 2. Amount of total protein synthesis was more increased in Drynariae Rhizomatreated group than in control group. In $5{\mu}g/ml$ Drynariae Rhizoma-treated group showed most prominent protein synthesis of the any other exrperimental group and control group. 3. Alkaline phosphatase activity also more increased in Drynariae Rhizomatreated group than control group. 4. Mineralized nodules in Drynariae Rhizoma-treated group were more than not in control group but also in PDGF-treated group. From the above results, Drynariae Rhizoma appeared to enhanced the proliferation, protein synthesis, alkaline phosphatase activity and cellular ability of mineralized nodule formation than PDGF. So that, we conclude that Drynariae Rhizoma enhances the activities of bone cells which have the important role on the periodontal regeneration and optimal application of Drynariae Rhizoma was thought to be useful as the means in bone regeneration.

• Journal of Periodontal and Implant Science
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• v.30 no.1
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• pp.77-91
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• 2000

The purpose of this study was performed to evaluate the effect of Armeniacae Semen extracts on human gingival fibroblasts and periodontal ligament cells in vitro. A experiment was done to evaluate the effect of Armeniacae Semen extracts in high glucose media. $400mg/d{\ell}$ glucose was added to the culture media of all groups. In control group, the cells($4.5{\times}10^4cells/ml$) were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum. In experimental groups, Armeniacae Semen extracts was added to the above culture media at the final concentrations of $1{\mu}g/m{\ell}$(Test group 1) and $l0{\mu}g/m{\ell}$(Test group 2). Then each group was tested for the rate of cell proliferation at 1, 2, 5 days, protein levels at 2, 5 days, and alkaline phosphatase activity at 2, 5 days. The results were as follows ; 1. Under the high glucose condition 1)As centration of Armeniacae Semen extracts increased, the rate of cell proliferation decreased significantly in test group 2 at 5 days in human gingival fibroblasts, but increased significantly in test group 2 at 5 days in human periodontal ligament cells(P<0.05). 2)In human gingival fibroblasts, test group 2 showed significantly decreased protein levels as compared to control group at 5 days. In periodontal ligament cells, test group 1 and 2 showed not significantly increased protein levels as compared to control group at 2, 5 days(P<0.05). 3)Alkaline phosphatase activity of human periodontal ligament cells increased as concentration of Armeniacae Semen extracts increased. The test group 1and 2 showed significant increase as compared to control group at 5 days(P<0.05). From the above results, Armeniacae Semen extracts appeared to enhance cellular activities including the rate of cell proliferation, protein levels and alkaline phosphatase activity of selectively human periodontal ligament cells in high glucose media. This study suggests that Armeniacae Semen extracts seem to be able to subside the inflammation of periodontal tissue and regenerate the destructed periodontal tissue.

• Animal cells and systems
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• v.11 no.1
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• pp.9-15
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• 2007

The naphthoquinone analog (2,3-dichloro-6,9-dihydroxy-1,4-naphtoquinone, NA) has an inhibitory effect on cdc25A protein phosphatase in vitro, which is responsible for G1/S transition during cell cycle. However, the exact mechanism inducing the growth inhibition is not understood. In this study, we investigated the regulatory mechanisms of growth arrest induced by NA, as a new potent inhibitor of cdc25A phosphatase, in human hepatocarcinoma SK-hep-1 cells. We found that NA induced the G1 arrest by perturbation of protein tyrosine dephosphorylation of Cdk2, which may be resulting from inhibition of cdc25A phosphatase. In addition, p21 was expressed in a p53-independent manner and participated in the NA-induced G1 arrest by inhibiting Cdk2 activity. Although the exact mechanism is not known, the p21 expression might be related to MAPK activation. From these results, we suggest that NA induces G1 arrest via inhibition of cdc25A and induction of p53-independent p21 expression in SK-Hep-1 cells.

• Journal of Radiation Protection and Research
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• v.18 no.2
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• pp.61-69
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• 1993

We examined various enzyme activity changes by intraperitoneal injection uranium in the carp liver. These enzyme activity changes can be used as biochemical indicators of internal exposure to uranium. The results were followings ; 1) Total protein concentration decreased by intraperitoneal injection in the carp liver. 2) Lysosomal acid pretense and ${\beta}-glucuronidase$ activities increased in the liver until sixth intraperitoneal injection of uranium, but Lysosomal acid phosphatase activities decreased in the liver until the sixth injection of uranium. 3) Alkaline phosphatase activities sharply increased and Glutamate oxaloacetate Transaminase activities steadily decreased in the liver until the sixth injection of uranium. 4) Creatine %kinase activities steadily decreased and malate dehydrogenase activities sharply decreased in the liver after the primary injection of uranium. Any malate dehydrogenase activities was not detected after sixth injection of uranium.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.7
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• pp.1033-1039
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• 2006

The metabolic response of Labeo rohita to thermal acclimation was assessed. Advanced fingerlings of L. rohita (average weight $31{\pm}1.4g$) were acclimated to 31, 33 and $36^{\circ}C$ compared with ambient temperatures ($26^{\circ}C$) for 30 days and different enzymes associated with stress response were estimated. Glycolytic enzyme-Lactate dehydrogenase, (LDH, E.C.1.1.1.27), TCA cycle enzyme-Malate dehydrogenase (MDH, E.C.1.1.1.37), Protein metabolizing enzymes-Aspartate amino transferase (AST, E.C.2.6.1.1) and Alanine amino transferase (ALT, E.C.2.6.1.2) of liver, gill and muscle, Gluconeogenic enzymes-Fructose 1,6 Bi phosphatase (FBPase, E.C. 3.1.3.11) and Glucose 6 phosphatase (G6Pase, E.C. 3.1.3.9) of liver and kidney were significantly (p<0.05) different with increasing acclimation temperatures. Heat Shock Protein-70 (HSP-70) was expressed in increasing intensity at 31, 33 and $36^{\circ}C$ but was not expressed at $26^{\circ}C$. Results suggest that higher acclimation temperatures enhance metabolism and L. rohita maintains homeostasis between $26-36^{\circ}C$ via an acclimation episode. Such adaptation appears to be facilitated by resorting to gluconeogenic and glycogenolytic pathways for energy mobilization and induction of HSPs.

• Journal of Sericultural and Entomological Science
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• v.39 no.2
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• pp.119-133
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• 1997

Isozyme was used to characterize general protein patterns of genetic relationships among 303 silkworm stocks preserved in National Sericultural and Entomology Research Institute, RDA. Six isozymes (esterase, acid phosphatase, alkaline phosphatase, amylase, glucose-6-phosphate dehydrogenase and sucrase) from hemolymph, midgut, and digestive juice were employed to construct dendograms(UPGMA method) using a polycrylamide gel electrophoresis. A cluster analysis revealed four major group, which were divided into several subgroups within each group, contained assemglages of Japanese and Chinese races. Especially, genetic differentiation in the first and second group was greatest rather than within Japanese and Chinese races repectively and was concordant with the hypothesis of phyletic sorting of initial variability in China many years ago. Hypothesized recent introgression between groups was also plausible, but the eviednce suggested bidirectional gene flow between the Chinese and the Japnaese lineages. Interpreting the results in light of evidence from the current study, the genetic diversity and relationship showed in Korean silkworm race, Hansammyun reflected early and independent evolution from the Chinese ancestor, limited addition of new variability and phyletic sorting within Korean peninsula more than 4,000 years.

• Journal of Cancer Prevention
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• v.22 no.4
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• pp.234-240
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• 2017

Background: Lung cancer is the leading cause of cancer-related death worldwide, for which smoking is considered as the primary risk factor. The present study was conducted to determine whether genetic alterations induced by radon exposure are associated with the susceptible risk of lung cancer in never smokers. Methods: To accurately identify mutations within individual tumors, next generation sequencing was conduct for 19 pairs of lung cancer tissue. The associations of germline and somatic variations with radon exposure were visualized using OncoPrint and heatmap graphs. Bioinformatic analysis was performed using various tools. Results: Alterations in several genes were implicated in lung cancer resulting from exposure to radon indoors, namely those in epidermal growth factor receptor (EGFR), tumor protein p53 (TP53), NK2 homeobox 1 (NKX2.1), phosphatase and tensin homolog (PTEN), chromodomain helicase DNA binding protein 7 (CHD7), discoidin domain receptor tyrosine kinase 2 (DDR2), lysine methyltransferase 2C (MLL3), chromodomain helicase DNA binding protein 5 (CHD5), FAT atypical cadherin 1 (FAT1), and dual specificity phosphatase 27 (putative) (DUSP27). Conclusions: While these genes might regulate the carcinogenic pathways of radioactivity, further analysis is needed to determine whether the genes are indeed completely responsible for causing lung cancer in never smokers exposed to residential radon.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1397-1401
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• 2005

Bone morphogenetic protein-4 (BMP-4) is a signaling homodimeric molecule that acts as a morphogen to influence cell fate in a concentration-dependent manner. The limited supply of a pure preparation of BMP-4, due to very low level of their expression in vivo, makes it difficult not only to study the biological activities of BMPs, but also to use them as a clinical tool. For a large-scale production of BMP-4, human BMP-4 cDNA was expressed in Chinese hamster ovary (CHO) cells by a recently development vector system, which confers position-independent stable expression of the foreign genes. The CHO cell line expressing recombinant human BMP-4 (rhBMP-4) at the level of $7\;{\mu}g/ml$ could be obtained after stepwise selection with methotrexate. This level of expression is about 70 times higher than those previously reported. The partially processed form of BMP-4 as well as mature form could be detected, when the aliquots of culture media were analyzed by Western blot. The glycosylation pattern and biological activity of the rhBMP-4 were determined by glycosidase treatment and the induction rate of alkaline phosphatase in mouse osteoblastic cells.

• Biomedical Science Letters
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• v.18 no.1
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• pp.1-9
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• 2012

Differentially expressed genes by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) were identified in order to evaluate them as dioxin-sensitive markers and crucial signaling molecules to understand dioxin-induced toxic mechanisms in human bronchial cells. Gene expression profiling was analyzed by cDNA microarray and ten genes were selected for further study. They were cytochrome P450, family 1, subfamily B, polypeptide 1 (CYP1B1), S100 calcium binding protein A8 (calgranulin A), S100 calcium binding protein A9 (calgranulin B), aldehyde dehydrogenase 1 family, member A3 (ALDH6) and peroxiredoxin 5 (PRDX5) in up-regulated group. Among them, CYP1B1 was used as a hallmark for dioxin and sharply increased by TCDD exposure. Down-regulated genes were IK cytokine, interferon-induced protein with tetratricopeptide repeats 1 (IFIT1), nuclease sensitive element binding protein 1 (NSEP1), protein tyrosine phosphatase type VI A, member 1 (PTP4A1), ras oncogene family 32 (RAB32). Although up-regulated 4 genes in microarray were coincided with northern hybridization, down-regulated 5 genes showed U-shaped expression pattern which is sharply decreased at lower doses and gradually increased at higher doses. These results introduce some of TCDD-responsive genes can be sensitive markers against TCDD exposure and used as signaling cues to understand toxicity initiated by TCDD inhalation in pulmonary tissues.

• Korean Journal of Microbiology
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• v.52 no.1
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• pp.10-17
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• 2016

Enzyme immunoassay to analyze specific binding activity of antibody to antigen uses horseradish peroxidase (HRP) or alkaline phosphatase (AP). Chemical methods are usually used for coupling of these enzymes to antibody, which is complicated and random cross-linking process. As results, it causes decreases or loss of functional activity of either antibody or enzyme. In addition, most enzyme assays use secondary antibody to detect antigen binding activity of primary antibody. Enzymes coupled to secondary antibody provide a binding signal by substrate-based color development, suggesting secondary antibody is required in enzyme immunoassay. Additional incubation time for binding of secondary antibody should also be necessary. More importantly, non-specific binding activity caused by secondary antibody should also be eliminated. In this study, we cloned AP isolated from Escherichia coli (E. coli) chromosome by PCR and fused to) hAY4 single-chain variable domain fragment (ScFv) specific to death receptor (DR4) which is a receptor for tumor necrosis factor ${\alpha}$ related apoptosis induced ligand (TRAIL). hAY4 ScFv-AP expressed in E. coli showed 73.8 kDa as a monomer in SDS-PAGE. However, this fusion protein shown in size-exclusion chromatography (SEC) exhibited 147.6 kDa as a dimer confirming that natural dimerization of AP by non-covalent association induced ScFv-AP dimerization. In several immunoassay such as ELISA, Western blot and immunocytochemistry, it showed antigen binding activity by color development of substrates catalyzed by AP directly fused to primary hAY4 ScFv without secondary antibody. In summary, hAY4 ScFv-AP fusion protein was successfully purified as a soluble dimeric form in E. coli and showed antigen binding activity in several immunoassays without addition of secondary antibody which sometimes causes time-consuming, expensive and non-specific false binding.