• BMB Reports
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• 제31권5호
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• pp.453-458
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• 1998

Human thrombopoietin (hTPO) was expressed to high levels in insect cells using the baculovirus expression system. Full-length hTPO cDNA containing a native signal peptide sequence was amplified by PCR from a human fetal liver cDNA library and cloned into the Autographa californica nuclear polyhedrosis virus (AcNPV) expression vector. Immunoblot analysis with antiserum against hTPO indicated that an approximately 55 kDa protein was produced in recombinant AcNPV infected insect cells. Recombinant hTPO was produced 4-fold higher in Trichoplusia ni (Tn5) cells than in Spodoptera frugiperda (Sf9) cells. with most of the hTPO produced in Tn5 cells secreted into the culture medium. Addition of tunicamycin in the culture medium resulted in the reduction of the size of hTPO to 35-38 kDa, and most of the protein remained within the cell. These results suggest that N-glycosylation of hTPO is required for the secretion of the protein into the culture medium in insect cells. hTPO produced in insect cells induced proliferation and maturation of megakaryocyte progenitors, indicating that it is in a biologically active form.

• 한국발생생물학회지:발생과생식
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• 제17권1호
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• pp.1-8
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• 2013

Osteoprotegerin (OPG) is a secreted glycoprotein that regulates bone resorption by inhibiting differentiation and activation of osteoclast, thereby potentially useful for the treatment of many bone diseases associated with increased bone loss. In this study, we designed a novel cDNA expression cassette by modifying the potent and mammary gland-specific goat ${\beta}$-casein/hGH hybrid gene construct and examined human OPG (hOPG) cDNA expression in transgenic mice. Six transgenic mice all successfully expressed hOPG in their milk at the level of 0.06-2,000 ${\mu}g/ml$. An estimated molecular weight of the milk hOPG was 55 kDa in SDS-PAGE, which is the same as a naturally glycosylated monomer. This hOPG expression was highly specific to the mammary glands of transgenic mice. hOPG mRNA was not detected in any organs analyzed except mammary gland. Functional integrity of milk hOPG was evaluated by TRAP (tartrate-resistant acid phosphatase) activity assay in bone marrow cell cultures. OPG ligand (OPG-L) treatment increased TRAP activity by two fold but it was completely abolished by co-treatment with transgenic milk containing hOPG. Taken together, our novel cDNA expression cassette could direct an efficient expression of biologically active hOPG, a potential candidate pharmaceutical for bone diseases, only in the mammary gland of transgenic mice.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2003년도 춘계 학술발표대회
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• pp.103-103
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• 2003

This study was carried out to investigate of the biologically active components in germinating Mung bean(Phaseolus radiata L.) and Buckwheat (Fagopyrum esculentum Moench) seeds. During the initial germination, germination ratio of 24 hours pre-soaking Mung bean and Buckwheat seeds were higher about 2∼3% than that of non-soaking. This experiment also was peformed to observe cytotoxic effect of the germinating seeds(germination length : 2, 5, 10mm) extracts against cancer cell lines including human lung carcinoma(Calu-6), human breast adenocarcinoma(MCT-7), human great intestine carcinoma(Caco-2) and human leukemia carcinoma(AML-2/WT). The growth of the cancer cells in medium containing Mung bean and Buckwheat extracts were significantly inhibited degree in proportion to the length of germination seeds, Especially, the results show that a significant shrinkage of Calu-6 cells was observed when the cells were exposed into extract of 10mm germination seeds in germinating Mung bean and Buckwheat seeds.

• 대한방사성의약품학회지
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• 제2권1호
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• pp.9-16
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• 2016

Carbon-11 is one of the most sensitive and desirable positron emission tomography radio-isotope, which offers the capacity to be incorporated, through a covalent bond, into biologically active molecules without altering their biological properties. Carbon-11 can be obtained from the cyclotron with two different chemical forms: $[^{11}C]CO_2$ and $[^{11}C]CH_4$. [$^{11}C$]Methyl iodide has been widely used as a highly reactive labelling precursor that can be applied to label carbon-11 with biologically active molecules via alkylation of N-, O-, or S-nucleophiles. A more recent and still challenging labeling method is transition metal mediated $^{11}C$-carbonylation. Advances in organic chemistry, radiochemistry and improved automated techniques greatly encourage researchers to develop more carbon-11 labelled radiotracers for molecular imaging studies. This mini-review will introduce a historical track of carbon-11 chemistry combining with examples and its role in near future.

• Journal of Ginseng Research
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• 제39권4호
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• pp.406-413
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• 2015

Background: Pathogenesis-related 10 (PR-10) proteins are small, cytosolic proteins with a similar three-dimensional structure. Crystal structures for several PR-10 homologs have similar overall folding patterns, with an unusually large internal cavity that is a binding site for biologically important molecules. Although structural information on PR-10 proteins is substantial, understanding of their biological function remains limited. Here, we showed that one of the PgPR-10 homologs, PgPR-10.3, shares binding properties with flavonoids, kinetin, emodin, deoxycholic acid, and ginsenoside Re (1 of the steroid glycosides). Methods: Gene expression patterns of PgPR-10.3 were analyzed by quantitative real-time PCR. The three-dimensional structure of PgPR-10 proteins was visualized by homology modeling, and docking to retrieve biologically active molecules was performed using AutoDock4 program. Results: Transcript levels of PgPR-10.3 expressed in leaves, stems, and roots of 3-wk-old ginseng plantlets were on average 86-fold lower than those of PgPR-10.2. In mature 2-yr-old ginseng plants, the mRNA of PgPR-10.3 is restricted to leaves. Ginsenoside Re production is especially prominent in leaves of Panax ginseng Meyer, and the binding property of PgPR-10.3 with ginsenoside Re suggests that this protein has an important role in the control of secondary metabolism. Conclusion: Although ginseng PR-10.3 gene is expressed in all organs of 3-wk-old plantlets, its expression is restricted to leaves in mature 2-yr-old ginseng plants. The putative binding property of PgPR-10.3 with Re is intriguing. Further verification of binding affinity with other biologically important molecules in the large hydrophobic cavity of PgPR-10.3 may provide an insight into the biological features of PR-10 proteins.

• Journal of Microbiology and Biotechnology
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• 제23권3호
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• pp.322-328
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• 2013

In this study, we show the expression and purification of the human recombinant farnesoid X receptor (FXR)- ligand binding domain (LBD) protein in E. coli using a double cistronic vector, pACYCDuet-1, as a soluble form. We describe here the expression and characterization of a biologically active $FXR-LBD_{(248-476)}$. When expressed in the influence of bacterial promoters ($P_{T7}$ and $P_{Tac}$) of the single cistronic expression vectors, the human recombinant $FXR-LBD_{(248-476)}$ was found to be totally insoluble. However, by using a double cistronic expression vector, we were able to obtain the human recombinant $FXR-LBD_{(248-476)}$ in a soluble form. To allow for biological activities, we have subcloned into the pACYCDuet-1 vector, expressed in E. coli cells at some optimized conditions, and purified and characterized the human recombinant active $FXR-LBD_{(248-476)}$ proteins using the fluorescence polarization assay. This suggests that the expression of FXR-LBD in a double cistronic vector improves its solubility and probably assists its correct folding for the biologically active form of the proteins. We suggest that this may represent a new approach to high expression of other nuclear receptors and may be useful as well for other classes of heterodimeric protein partners.

• Journal of Microbiology and Biotechnology
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• 제27권7호
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• pp.1281-1287
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• 2017

Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of $EC_{50}$ of 2.93 ng/ml.

• 생약학회지
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• 제15권2호
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• pp.85-90
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• 1984

The whole plant of Duchesnea indica (Andr.) Focke (Rosaceae) which has been used in folklore in treating amenorrhea, inflammation, fever and traumatic injuries, in detoxifying and breaking up clots, was studied. The pharmacological test showed that the water extract had estrogenic and histaminergic actions, but no antiserotonin action. As a result of systematic separation in order to detect the active compounds revealing the estrogenic effect, the active compounds were found in the ether fraction. TLC of the ether fraction revealed 8 spots. Among them three major spots (Rf=0.54, 037, 0.31) were separated by preparative TLC. Some chemical properties of those major spots suggested that they were phenolic compounds, but that they were neither linoleic acid nor ${\beta}-sitosterol$ previously reported.

• 생약학회지
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• 제21권3호
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• pp.223-234
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• 1990

Hepatoprotective activities of 26 traditional Korean herbal prescriptions described in Donghibogam were evaluated in vivo and in vitro screening system. Twenty one prescriptions appeared to be active. Six prescriptions among them showed highly significant hepatoprotective activities; Shihogyegy-Tang(柴胡桂枝湯), Soshiho-Tang(小柴胡湯), Shiho plus Younggolmoryo-Tang(柴胡加龍骨牡蠣湯), Yinjinho-Tang(茵蔯蒿湯), Yinsam-Tang(人蔘湯) and Shashim-Tang(瀉心湯). Shihogyegy-Tang was most active. They reduced the release of transaminases from dissociated hepatocytes by thioacetamide, and serum transaminases and alkaline phosphatase activities of rat with carbontetrachloride-induced hepatitis. They inhibited significantly lipid peroxidation and cellular fatty change of liver by carbontetrachloride.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.334.2-334.2
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• 2002

As the part of our continuing study for antihyperlipidemic agents from the herbal medicinal resources. we examined the possibility of the ethylacetate fraction of Alpinia officinarum water extract in vitro and in vivo. We isolated some compounds from the ethylacetate fraction of Alpinia officinarum and measured their antihyperlipidemic activities. The active components isolated by silicagel column lmproved serum TG. HDL and LDL level in corn oil feeding and triton WR-1339 induced hyperlipidemic mice. The most active compound was 3-Methylethergalangin. Consequently. these biologically active gerbal functional foods could be used for preparing the healthy food which might diabetas. hyperlipidemia and other disease.