• Journal of Plant Biotechnology
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• v.32 no.1
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• pp.51-55
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• 2005

Chinese leymus (Leymus chinensis Trin.) is a perennial grass that is widely distributed at high pH sodic and arid soil in the northeastern Asia. An efficient regeneration system was established through somatic embryogenesis of mature seeds to understand its high adaptability to harsh environmental conditions on the basis of molecular biology. The calli were efficiently induced (about $70\%$) from mature seeds on MS medium supplemented with $1.5\;\cal{mg/L}$ 2,4-D. Somatic embryos were formed from the surface of embryogenic callus on MS medium supplemented with $2.0\;\cal{mg/L}\;kinetin\;and\;0.5\;\cal{mg/L}$ NAA after 3 weeks of culture. Roots were induced from the shoot when transferred to MS medium without plant growth regulator for 1 week. Plant regeneration rate was $36\%$ and regenerated plantlets were grown to normal mature plants in pot. An efficient plant regeneration system in this study will be useful for molecular breeding of L. chinensis.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.2
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• pp.227-233
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• 2004

Chronic exposure to solar radiation, particularly ultraviolet (UV) light, causes a variety of adverse reactions on human skin, such as sunburn, photoaging and photocarcinogenesis. Free radicals and reactive oxygen species (ROS) caused by UV exposure or other environmental facts play critical roles in cellular damage. And, repeated-UV irradiation activated the expression of the matrix metalloproteinase (MMP) and induced skin irritation. Therefore, the development of effective and safe photoprotectants that can reduce and improve the skin damage has been required. The purpose of this study was to investigate the photo-protective effect of several chinese medical plants (Juniperus chinensis) on the UV -induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. Fluorometric assays of the proteolytic activities of MMP-1 (collagenase) were performed using fluorescent collagen substrates. UVA induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. Expression of prostaglandin E$_2$ (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay(EIA) using PGE$_2$ monoclonal antibody. In the human skin we tested anti-irritation effect on the SLS-induced damage skin after appling the extract containing emulsion. We found that Juniperus chinensis extract had potent radical scavenging effect by 98% at 100$\mu\textrm{g}$/mL. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25$\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. In the test of proinflammatory cytokines of human keratinocytes Juniperus chinensis extract decreased expression of interleukin 6 about 30%. The amount of PGE$_2$ by HaCaT keratinocytes was significantly increased at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB (p < 0.05). At the concentrations of 3.2-25$\mu\textrm{g}$/mL of this extract, the production of PGE$_2$ by HaCaT keratinocytes (24 h after 10mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p < 0.05). In SLS-induced skin irritation model in vivo, we found to reduce skin erythema and improve barrier recovery after appling Juniperus chinensis extract containing emulsion when compared to irritated non-treated and placebo-treated skin. Our results suggest that Juniperus chinensis extract can be effectively used for the prevention of UV and SLS-induced adverse skin reactions and applied as anti-aging and anti-irritation cosmetics.

• The Journal of Korean Society of Virology
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• v.26 no.2
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• pp.215-225
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• 1996

Aseptic meningits, an acute inflammation of the meninges, is a common illness during childhood. Virus is the most important cause of aseptic meningitis. Especially enterovirus causes approximately above 85% of all cases of aseptic meningitis. In 1993, there was a big epidemic of aseptic meningitis by ECHO 9 and ECHO 30 viruses. And ECHO 3 virus was isolated as a causative agent of aseptic meningitis in 1994. This study was aimed to detect the causative agent of aseptic meningitis in 1995 and to analyze the 5'-noncoding region which was used to detect virus. Virus was isolated from 87 stools and cerebrospinal fluid specimens of the patients by cultured RD and HEp-2 cell. Neutralizing antibody tests using enterovirus serum pool were performed on the specimens with cytopathic effect. 3 of ECHO 7 viruses and 5 of Coxsackie B3 viruses were isolated from stool specimens and 1 of ECHO 7 and Coxsackie B3 mixed type was confirmed from cerebrospinal fluid specimens. RNA was isolated from the culture supernatants of infected cells and general primers were selected in highly conserved part of the 5'-noncoding region of the enteroviral genome for RT-PCR. PCR product from this virus showed a 152bp band on gel electrophoresis. Sequence of obtained DNA was compared with prototype sequences by accessing to the Genebank database. 5'-noncoding region of isolated Coxsackie B3 virus, which has point mutations in nucleotide sequence positions 493, 497, 502, 523, was closely related to that of polio virus type 1, Mahoney strain. In case of isolated ECHO 7 virus, nucleotide has been changed from cytosine to thymine at position 581 and from thymine to cytosine at position 583. We concluded the causative agents of the outbreak of aseptic meningitis during June to July in 1995 were both ECHO 7 and Coxsackie B3 virus, and the primer used in this study could allow a rapid diagnosis of enteroviruses by PCR.

• Journal of Microbiology and Biotechnology
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• v.31 no.7
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• pp.933-941
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• 2021

Ginsenoside Rg4 is a rare ginsenoside that is naturally found in ginseng, and exhibits a wide range of biological activities including antioxidant and anti-inflammatory properties in several cell types. The purpose of this study was to use an in vivo model of hair follicle (HF)-mimic based on a human dermal papilla (DP) spheroid system prepared by three-dimensional (3D) culture and to investigate the effect of Rg4 on the hair-inductive properties of DP cells. Treatment of the DP spheroids with Rg4 (20 to 50 ㎍/ml) significantly increased the viability and size of the DP spheres in a dose-dependent manner. Rg4 also increased the mRNA and protein expression of DP signature genes that are related to hair growth including ALP, BMP2, and VCAN in the DP spheres. Analysis of the signaling molecules and luciferase reporter assays further revealed that Rg4 induces the activation of phosphoinositide 3-kinase (PI3K)/AKT and the inhibitory phosphorylation of GSK3β, which activates the WNT/β-catenin signaling pathway. These results correlated with not only the increased nuclear translocation of β-catenin following the treatment of the DP spheres with Rg4 but also the significant elevation of mRNA expression of the downstream target genes of the WNT/β-catenin pathway including WNT5A, β-catenin, and LEF1. In conclusion, these results demonstrated that ginsenoside Rg4 promotes the hair-inductive properties of DP cells by activating the AKT/GSK3β/β-catenin signaling pathway in DP spheres, suggesting that Rg4 could be a potential natural therapy for hair growth.

• Development and Reproduction
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• v.6 no.2
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• pp.97-104
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• 2002

In most mammals, mature oocyte-cumulus complexer(OCCs) ovulate into the oviduct where fertilization by sperm takes place. However, the complex that fail to fertilize eventually undergoes degeneration while they reside in the oviduct. Yet there is no blown mechanism how both oocyte and cumulus cells degenerate. Using human follicular fluid (hFF), bovine oviductal tissue extract (BOX) and mouse OCC, the present study aimed to find how the oviduct influence the viability of the oocyte and cumulus cells in vitro. There was no difference of oocyte maturation rate between the control and BOX-treated groups. However, there was a significant difference in the survival of cumulus cells between two groups. Cumulus cells cultured in the presence of hFF alone underwent initially expansion and then they formed monolayer in the culture dish. Even after 72 hr, they proliferated well and showed fibroblast-like morphology. Cumulus cells cultured in the presence of both hFF and BOX also expanded after 24 hr, however, after 72 hr culture, they eventually detached and degenerated. Cumulus cells cultured in the BOX alone gave a similar drastic result. When the cumulus cells cultured in the presence of BOX were stained with DAPI, their nuclei showed partial condensation and fragmentation. After detailed analysis of these cells by TUNEL assay, many nuclei of them exhibited well stained spots indicating the signs of apoptosis. Based upon these observations, it is suggested that BOX might possess a factor that leads mouse cumulus cells to undergo apoptosis in vitro.

• The Korean Journal of Mycology
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• v.46 no.2
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• pp.161-176
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• 2018

Fungal ${\beta}$-glucan, known to have immunostimulatory and antitumor activities, can be recognized by host immune cells as one of the pathogen-associated molecular patterns (PAMPs). Although there are several reports on the diverse immunostimulatory activities of ${\beta}$-glucan, little is known about the intracellular signal transduction of ${\beta}$-glucan. Stimulation of RAW264.7 macrophage cells with ${\beta}$-glucan from Ganoderma lucidum induced the expressions of dectin-1, toll-like receptor 2 (TLR2), TLR4, and TLR6 at the transcription stage. Treatment with ${\beta}$-glucan also induced inflammatory mediators such as macrophage inflammatory proteins (MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-$1{\gamma}$, interleukin (IL)-$1{\beta}$, and tumor necrosis factor (TNF)-${\alpha}$. Treatment of the cells with polymyxin B, an inhibitor of lipopolysaccharides (LPS), blocked the induction of inflammatory mediators in LPS- or ${\beta}$-glucan-stimulated systems. Pretreatment of the cells in our cell culture system with LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, or U0126, a mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) kinase (MEK)1/MEK2 inhibitor, led to a reduction in the induction of inflammatory mediators in a concentration-dependent manner. These results show that stimulation of the macrophage cells by ${\beta}$-glucan induced the expressions of both dectin-1 and TLRs. We also found that the PI3K/Akt and MEK pathways were involved in the induction of inflammatory mediators in macrophage cells during intracellular signal transduction of ${\beta}$-glucan.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.1
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• pp.20-26
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• 2016

This study investigated the fermentation effect of Taraxacum platycarpum Dahlst. leaf extracts using Hericium ernaceum mycelia to test antioxidant and anti-inflammatory activities in vitro. The antioxidant activities of fermented or non-fermented extracts of T. platycarpum leaves were determined by 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, ferric reducing antioxidant power, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activity. The leaf extract of T. platycarpum showed higher antioxidant activity than extract of fermented leaves. However, ethanolic extract of fermented T. platycarpum leaves decreased levels of nitric oxide production and pro-inflammatory cytokines such as interleukin-6 and tumor necrosis factor-${\alpha}$ in lipopolysaccharide-stimulated RAW 264.7 cells. Moreover, fermented leaf extract suppressed protein expression of inducible nitric oxide synthase in RAW 264.7 cell culture. Therefore, the enhanced anti-inflammatory activity of ethanolic extracts of fermented T. platycarpum leaves might be attributed to the molecular conversion of leaf ingredients during fermentation and the active ingredients might have specific affinity with ethanol during extraction.

• Korean Journal of Pharmacognosy
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• v.5 no.2
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• pp.111-124
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• 1974

The radioactive compound sodium $acetate-U-C^{14}\;(C^{14}-acetate)$ was administered to two- and four-year-old July and September American ginseng (Araliaceae, Panax quinquefolium L.) plants and cuttings. The $C^{14}-acetate$ uptake was approximately 99%. The autoradiochromatograms suggest that the saponins isolated by preparative thin-layer chromatography contained impurities, especially those isolated from the leaf and stem extracts. The root and fruit methanol extracts yielded relatively pure saponins. The large amounts of panaquilin B and its proximity to panaquilin C on preparative thin-layer plates resulted in some admixing. The average concentration (% plant dry weight) of semi-purified saponins were high in the leaves (13.8%), as compared to fruits (9.8%), stems (7.9%) and roots (6.3%). The average percentage of $C^{14}-acetate$ incorporation into panaquilins was 4.8%. The average percentage of $C^{14}-acetate$ incorporation into panaquilins B and C was higher (1.40% and 1.13%, respectively) than that into panaquilins C, (d), G-1 and G-2 (0.75%, 0.65%, 0.13% and 0.53%, respectively). Panaquilin synthesis may be depending upon the part, collection period and age of the plant. The average percentage of $C^{14}-acetate$ incorporation into panaquilin B is high in roots (0.58%) and stems (0.48%); that into panaquilins C and (d) high in leaves (0.40% and 0.45%, respectively); and that into panaquilin E high in roots and leaves (0.55% and 0.50%, respectively). Panaquilin G-2 was synthesized in all parts of plants. The panaquilins appear to be biosynthesized more actively in July than September (exception-panaquilin G-1). Panaquilins B, C and G-1 may be biosynthesized more actively in four-year-old plants and panaquilins (d) and E more actively in two-year-old plants. The results from expectance with cuttings suggest that the panaquilins are synthesized de novo in the above-ground parts of ginseng plants, and that panaquilin G-1 may be synthesized de novo in the leaf. It is known from the tissue culture studies that panaquilins are produced by leaf, stem and root callus tissues and cailus-root cultures of American and Korean ginseng plants. Panaquilins may actively be synthesized de novo in most any cell or organ of the ginseng plants. It was verified that $C^{14}-acetate$ was incorporated into the panaxadiol portions of the panaquilins of two-year-old plants (sp. act. 0.56 mmcCi/mg) and four-year-old plants $(sp.\;act.\;0.54\;m{\mu}Ci/mg)$.

• The Journal of Natural Sciences
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• v.4
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• pp.103-142
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• 1991

These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.31 no.4 s.54
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• pp.349-357
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• 2005

New antioxidative substances for cosmeceuticals were screened from natural resources such as microbial metabolites, mushrooms, and medicinal plants. Four antioxidants were isolated from the fungal metabolite of Eupenicillium shearii and their structures were determined to be new phenolic compounds. The compounds were designated as melanocins A, B, C, and D. Melanocins $A{\sim}D$ exhibited free radical scavenging activity on DPPH and superoxide with $EC_{50}$ values of $21{\sim}94\;and\;7{\sim}84{\mu}M$, respectively, which were stronger activity than those of ${\alpha}-tocopherol$ and BHA. Melanocin A showed anti-wrinkle effects on the UV-irrated hairless mouse skin. A novel hispidin antioxidative compound designated as inoscavin A was isolated from the fruiting body of the mushroom, Inonotus xeranticus. Inoscavin A scavenged superoxide radical with $EC_{50}$ values of $0.03{\mu}g/mL$, and inhibited rat liver microsomal lipid peroxidation with $EC_{50}$ values of $0.3{\mu}g/mL$. Benzastatins $A{\sim}G$, the novel antioxidants isolated from the culture of Streptomyces nitrosporeus showed potent lipid peroxidation inhibitory activity with $EC_{50}$ values of $3{\sim}30{\mu}M$. A cyclopentene compound with strong hypopigmentary effect was isolated from the fungal metabolite of Penicillium sp. and identifed as terrein. Terrein significantly reduced melanin levels in a melanomacyte cell line, Mel-Ab. It showed 10 times stronger activity than kojic acid, but exhibited no cytotoxic effect even in $100{\mu}M$. It was suggested that terrein reduced melanin synthesis by reducing tyrosinase production by MITF down-regulation.