• Current Research on Agriculture and Life Sciences
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• v.21
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• pp.11-16
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• 2003

Lines bred for resistance to Phytophthora capsici by incorporation of resistance to P. capsici in PI201234 or CM334 into 'Subi' and 'Chilseong', land races in Youngyang, and 'Punggak', a land race in Cheongdo in Gyeungbuk province, and lines bred for fortification of one of them above with resistance to viral complex, and tolerant selections from another landrace collection from Punggak (KC268) were evaluated for resistance to P. capsici by inoculation at seedling stage. Almost all the breeding lines showed high level of resistance to P. capsid and selections from KC268 showed tolerance or moderate resistance to P. capsid. The selected plants were grown in a net cage in an outdoor nursery for seed production. Utilization of the lines in breeding was discussed.

• Korean Journal of Pharmacognosy
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• v.30 no.3
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• pp.238-243
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• 1999

This study was undertaken to test for anti-hepatitis B virus (HBV) activity of the aqueous extracts prepared from 9 medicinal plants of Korea (Cornus officinalis, Caesalpinia sappan, Rubus coreanus, Lycium chinense, Artemisia capillaris, Isatis tinctoria, Phyllanthus urinaria, Lysimachia christinae, Lonicera japonica). Aqueous extracts were tested for cytotoxicity and assayed for inhibition of HBV replication by measurement of HBV DNA and surface antigen (HBsAg) levels in the extracellular medium f HepG2 2.2.15 cells. The extract from Rubus coreanus, Artemisia capillaris, Phyllanthus urinaria decreased the levels of extracellular HBV virion DNA at concentrations ranging from 128 to $256\;{\mu}g/ml$ and inhibited the production fo HBsAg dose-dependently without showing cytotoxicity. Our findings suggest that these three hebal medicinal plants may have potential to develop as specific anti-HBV drugs in the future.

• Biomolecules & Therapeutics
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• v.6 no.2
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• pp.139-144
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• 1998

Abstract -Phyllanthus ussuriensis which belongs to Euphorbiaceae has been used as a component of Korean traditional remedy against hepatitis and jaundice. Aqueous methanolic extract of P. ussuriensis was tested for antiviral activity of hepatitis B virus (HBV) in HepG2 2.2.15 cells which were derived through transfection of cloned HBV DNA into HepG2 human hrpatoblastoma cells. P. ussuriensis extract decreased the levels of extracellular HBV virion DNA and inhibited HBV replication at concentrations ranging from 50 to 500$\mu$g/ml. Partitioning of water suspension of the extract revealed that the activity mainly resides in the EtOAc fraction. Consecutive purification of the EtOAc fraction by silica-gel column chromatography resulted in the two active anti-HBV fractions. Southern blot analysis shows that the action mechanisms or active site of the two fractions seems to be different in terms of their inhibition of HBV replication steps.

• Journal of Plant Biotechnology
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• v.44 no.4
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• pp.401-408
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• 2017

Herein, we report the meristem-tip culture from dormant buds of grape 'Kyoho' single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2-5 leaf primordia, and 1-4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at $4^{\circ}C$). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.

• Journal of Life Science
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• v.26 no.7
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• pp.764-771
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• 2016

Extracts from Artemisia annua Linné (AAE) have been known to possess various functions, including anti-bacterial, anti-virus, and anti-oxidant effects. However, the mechanism of those effects of AAE is not well-known. The aim of this study was to analyze the inhibitory effects of AAE on cell proliferation of the human hepatoma cell line (Hep3B) and to examine its effects on apoptosis. Activation by phosphorylation of Akt is cell proliferation through the phosphorylation of TSC2, mTOR, and GSK-3β. We suggested that AAE may exert cancer cell apoptosis through Akt/mTOR/GSK-3β signal pathways and mitochondria-mediated apoptotic proteins. For this, we examined the effects of extracts of AAE on cell proliferation according to treatment concentration. Treatment with AAE not only reduced cell viability, but also resulted in the induced release of lactate dehydrogenase (LDH). These results were determined with a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and a lactate dehydrogenase (LDH) assay. Furthermore, we determined the effects of apoptosis through Hoechst 33342 staining, annexinⅤ-propidium iodide (PI) staining, 5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide (JC-1) staining, and Western blotting. Our study showed that the treatment of liver cancer cells with AAE resulted in the inhibition of Akt, TSC2, GSK-3β-phosphorylated, Bcl-2, and pro-caspase 3 and the activation of Bim, Bax, Bak, and cleaved PARP expressions. These results indicate that AAE induced apoptosis by means of a mitochondrial event through the regulate of Akt/mTOR/GSK-3β signaling pathways.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.186-192
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• 2008

For the development of specific and effective basic genetic materials to inhibit replication of hepatitis C virus (HCV), HCV genome-targeting trans-splicing aptazyme, which activity is allosterically regulated by a specific ligand, was developed. The aptazyme was designed to be comprised of sequence of RNA aptamer to the ligand, communication module sequence which can transfer structural transition for inducing ribozyme activity upon binding the ligand to the aptamer, and trans-splicing ribozyme targeting +199 nt of HCV IRES. Especially, when the aptamer and the communication module was inserted at both P6 and P8 catalytic domain of the specific ribozyme, allosteric activity of the aptazyme was the most induced. The aptazyme was shown to induce activity of trans-splicing reaction specifically and efficiently only in the presence of the specific ligand, but neither in the absence of any ligand nor in the presence of control ligand. This aptazyme can be used as a specific and effective genetic agent against HCV, and a tool for the isolation of anti-HCV lead compounds.

• Korean journal of applied entomology
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• v.31 no.4
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• pp.396-415
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• 1992

To investigate identification of species distribution state, population density of plant parasitic nematodes and damaged state by parasitism of nematodes with soil samples taken from major cultivating area of medicinal herbs in Kyungbuk districts are summarized as follow; total species belonging to genera in families were identified from 43 different medicinal herbs. The dominant nematode species on medicinal herbs was Meloidogyne hapla by 55.6% of field infection. Damaged state by parasitizm of Root-knot nematode, M. hapla, M. incognita are severed 15 medical plant except P. japonica, infected field were in 54.5~88.0%, and highered in population densities of 2nd larvae, Gall, Egg sac. Percentage of yield-decrease were 57.8% in A. gigas, 49.1% in P.japonica. Lighten effect of M. spp. by cropping system were lowered in population density in Rice+Paeony than Paeony+Paeony in paddy field, and also lowered in Sesame+Paeony or one year fallowing after harvest Paeony than Red pepper+Pseony, Paeony+Paeony in upland field.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.283-292
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• 2009

Objective: Human embryonic stem cells (hESCs) can proliferate indefinitely and differentiate into all kinds of cell types in vitro. Therefore, hESCs can be used as a cell source for cell-based therapy. Transduction of foreign genes to hESCs could be useful for tracing differentiation processes of hESCs and elucidation of gene function. Thus, we tried to introduce enhanced green fluorescent protein (eGFP) gene to hESCs, XX and XY cell lines in this study. Methods: Lentivirus containing eGFP was packaged in 293T cells and applied to hESCs to transduce eGFP. Expression of transduced eGFP was evaluated under the fluorescence microscope and eGFP positive population was analyzed by FACS. Expression of undifferentiation state markers such as Oct4, Nanog, SSEA4 and Tra-1-81 was examined by RT-PCR and/or immunofluorescence in eGFP-hESCs after transduction. In addition, the ability of eGFP-hESCs to form embryoid bodies (EBs) was tested. Results: eGFP was successfully transduced to hESCs by lentivirus. eGFP expression was stably maintained up to more than 40 passages. eGFP-hESCs retained expression patterns of undifferentiation state markers after transduction. Interestingly, disappearance of transduced eGFP was notably observed during spontaneous differentiation of eGFP-hESCs. Conclusion: We established eGFP expressing hESC lines using lentivirus and showed the maintenance of undifferentiation characteristics of these eGFP-hESCs. This reporter-containing hESCs could be useful for tracing the processes of differentiation of hESCs and other studies.

• Clinical and Experimental Pediatrics
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• v.50 no.10
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• pp.1011-1017
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• 2007

Purpose : We tried to assess the optimal conditions to improve low transduction efficiency and their effect on target cells. Methods : Cultured NIH 3T3 cells were incubated with retroviral vectors bearing an enhanced green fluorescent protein (eGFP) gene. We varied the ratio of viral vectors to target cells (1:1-1:8) and the number of transfections (${\times}1$, ${\times}2$), and compared transduction efficiencies. Also, the effects of polybrene on transduction efficiency and viability of target cells were assessed. Transduction of the eGFP gene was evaluated by observing NIH 3T3 cells under a fluorescence microscope and efficiencies were measured by the percentage of eGFP positive cells using FACscan. Results : As the ratio of retroviral vectors to target cells increased, transduction efficiency was greatly improved, from 7% (1:1) to 38% (1:4). However, transduction efficiency did not increase any more when the ratio increased from 1:4 to 1:8. Cells transfected twice showed higher transduction efficiencies than cells transfected once, at a ratio of 1:8. The eGFP gene transduced to NIH 3T3 cells sustained its expression during repeated passages. However, after the third passage (day 9), the percentage of eGFP positive cells began to decline. The degree of this decline in eGFP expression was lower in cells transfected twice than in cells transfected once (P<0.05). The addition of polybrene did not have any toxic effect on NIH 3T3 cells and greatly increased transduction efficiency (P=0.007). In addition to vector component, transduction efficiency was very sensitive to culture confluence. Cells cultured and transfected in 24-well plate showed higher transduction efficiency, although cells cultured in 6- well plate proliferated more (P=0.024). Conclusion : Our data could be used as a basis for retrovirus-based gene therapy. Further study will follow using human cells as target cells.

• Investigative Magnetic Resonance Imaging
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• v.16 no.1
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• pp.47-54
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• 2012

Purpose : This study was performed to evaluate the characteristics of rat mesenchymal stem cells (RMSCs) transduced with human ferritin gene and investigate $in$ $vitro$ MRI detectability of ferritin-transduced RMSCs. Materials and Methods: The RMSCs expressing both myc-tagged human ferritin heavy chain subunit (myc-FTH) and green fluorescence protein (GFP) were transduced with lentiviurs. Transduced cells were sorted by GFP expression using a fluorescence-activated cell sorter. Myc-FTH and GFP expression in transduced cells were detected by immunofluorescence staining. The cell proliferative ability and viability were assessed by MTT assay. The RMSC surface markers (CD29+/CD45-) were analyzed by flow cytometry. The intracellular iron amount was measured spectrophotometically and the presence of ferritin-iron accumulation was detected by Prussian blue staining. $In$ $vitro$ magnetic resonance imaging (MRI) study of cell phantoms was done on 9.4 T MR scanner to evaluate the feasibility of imaging the ferritin-transduced RMSCs. Results: The myc-FTH and GFP genes were stably transduced into RMSCs. No significant differences were observed in terms of biologic properties in transduced RMSCs compared with non-transduced RMSCs. Ferritin-transduced RMSCs exhibited increased iron accumulation ability and showed significantly lower $T_2$ relaxation time than non-transduced RMSCs. Conclusion: Ferritin gene as MR reporter gene could be used for non-invasive tracking and visualization of therapeutic mesenchymal stem cells by MRI.