This study was estimated for cabbage aphid, Brevicoryne brassicae in Brassica campestris L. var. rapa (L.) Hartm. in order to institute of Economic injury levels(EILs). B. brassicae was innoculated on April 29, in differently 0, 5, 10, 20 and 40 adults per ten plant, respectively. After inoculated of B. brassicae, the density was increased until harvest ing gradually in all plots except non innoculate plot. and Higher inoculation density were increased higher than lower inoculation density. Percentage of damage leaf was higher in plots with higher initial aphid density than in plots with lower initial aphid density. And the leaf weight of commodity were decreased in higher initial aphid density. The decreasing rates of leaf weight of commodity was increased with increasing initial aphid density. The relationship between initial B. brassicae densities and the decreasing rates of leaf weight of commodity was well described by a linear regression, Y=0.8416X-3.5147, R2=0.94. Based on the relationship, the number of adults per 10 plant which can cause 5% loss of yield was estimated to be approximately 10.1. And EILs was estimated to be approximately 1.0 adults/plant in late April.

A full genomic DNA microarray technique was employed to investigate the effects of Dongchunghacho on aortal and hepatic gene expression in apolipoprotein E knockout mice fed a high-fat/high-cholesterol diet. Male 8- week - old ApoE-/- mice were randomly divided into two groups, control(high cholesterol group; HC) and supplementation of Dongchunghacho (SD). All of the mice were fed a high-fet/high cholesterol diet with or without Dongchunghacho supplemented by 1% for 6 weeks. At first, lipid profile of the Dongchunghacho was measured by biochemical analysis. No differences were observed in serum triglyceride and total cholesterol levels between the two groups. Antigenotoxic effect of the Dongchunghacho was measured by the single cell gel electrophoresis assay (Comet assay) and quantified as % fluorescence in tail. Dongchunghacho supplementation decreased significantly leukocytic DNA damage and also there was a tendency of reduction in hepatic DNA damage in Dongchunghacho group compared with the control group. In up regulated genes in liver and aorta of the mice, genes with 0 to 2- fold difference in expression level between the two group (HD and SD) was very much more in liver than in aorta, on the contrary, those with 2-fold to 16-flod difference increased greatly rather in aorta than in liver. Also, almost the same results were observed in down regulated genes in liver and aorta between the two groups. These results suggested that supplementation of Dongchunghacho might be helpful in preventing leukocytic DNA damage induced by high fat diet, and has a more crucial roles in aortal gene expression.

The completely sequenced mitochondrial genome of the brown marmorated stink bug, Halyomorpha halys, is a circular molecule of 16,518 bp with a total A+T content of 76.4%. Nucleotide composition and codon usage of this genome are near the means observed in other 12 hemipteran mitochondrial genomes; however, the initiation codon for CO1 gene appears to be TTG, dissimilar to what has been seen in the 12 mitochondrial genomes. In this genome, the A+T rich region between srRNA and tRNAIle gene includes two extensive repeat regions, in which each region includes 4 and 12 tandem repeats of a 73 bp sequence, respectively. The gene content, order, and structure of the H. halys mitochondrial genome are consistent with that of Triatoma dimidiata, belong to the same suborder Heteroptera, but different from two suborders, Auchenorrhynca and Sternorrhyncha, including various gene rearrangements. Analyzing phylogenetic relationship and comparing gene order and content of the 13 hemipteran mitochondrial genomes of three suborders, Heteroptera, Auchenorrhynca, and Sternorrhyncha, supported the morphology-based current hypothesis that both Auchenorrhynca and Sternorrhyncha are a monophyletic group.

Wntsignaling is involved in the normal development and tumorigenesis via epithelial- mesenchymal transition (EMT). init iated by down-regul ation of E-cadherin by the transc ription factor Snail. Wnt signaling inhi bits Sna il phosph o rylation t hrough Axin2-dependent pathway that sustains nuclear accumul ation 0 1' Snail by driving CSK3ß nucleocytoplasmic export then consequently increases Snail protein levels and induces an EMT However. the roles of Wnt and Axin expression and their functional implication on Snail dependent EMT program a re not clear du ring the multistep carcinogenic process. We examined that canonical Wnt signaling engagingmul t istep carcinogenic process of uterine cervical cancer through Wnt-Axin2-Snail axis. In nonnal cervi cal mucosa, Wntl. Wnt3a. and Axin2 mRNA expression were locali zed in basal cell layer suggesting that canonical Wnt is required for maintenance of self-renewal program of cervica l epi theli al cells. With progression to cervical int r aepithelial neoplasia (CIN) and carcinoma. Wntl, Wnt3a‘ Axin2, and Snail expression were gradually increased in patient samples suggesting that canonical Wnt pathway is involved in earl y step of carcinogenesis in uterine cervix. LRP6 and Axin2 transfected cells showed the highly increased nuclear Snai l resul ted from dec reased level 0 1' nu clear GSK3ß , indicating that LRP6- Axin2 serves to stabili ze Sna il protein levels and susLains iLs nllclear acc llrnulation by driv ing GSK3ß . RNA interference of Axin2 and Snail on SiHa cells relieved E-cadherin proximal promotel‘ activity and block the in 띠 vo chorioalantoic membra ne ln VaSlOn These results suggest the canon ical Wnt signa ling regul ating Axin2-GSK3ß compartmentalization may important for stabi li zation of E- cad herin repressor Snail during the multistep carcinogen ic process of uteri ne cervix. It may lead to not only tracing the proper biomarker 0 [' ca ncer progression‘ but a lso the development oJ new targets for therapeutic intervention in cancer

The stem cell research is emerging as a cutting edge topic for a new treatment for many chronic diseases. Recently, dental stem cell would be possible for regeneration of tooth itself as well as periodontal tissue. However, the study of the cell characterization is scarce. Therefore, we performed the genetic profiling and the characterization of mouse fetus/neonate derived dental tissue and cell to find the identification during dental development. We separated dental arch from mandibles of 14.5 d fetal mice and neonate 0 d under the stereoscope, and isolated dental cells primarily from the tissues. Then, we examined morphology and the gene expression profiles of the primary cells and dental tissues from fetus/neonate and adult with RT-PCR. Primary dental cells showed heterogeneous but the majority was shown as fibroblast-like morphology. The change of population doubling time levels (PDLs) showed that the primary dental cells have growth potential and could be expanded under our culture conditions without reduction of growth rate. Immunocytochemical and flow cytometric analyses were performed to characterize the primary dental cell populations from both of fetus (E14.5) and neonate. Alpha smooth muscle actin (α-SMA), vimentin, and von Willebrand factor showed strong expression, but desmin positive cells were not detected in the primary dental cells. Most of the markers were not uniformly expressed, but found in subsets of cells, indicating that the primary dental cell population is heterogeneous, and characteristics of the populations were changed during culture period. And mesenchymal stem cell markers were highly expressed. Gene expression profile showed Wnt family and its related signaling molecules, growth factors, transcription factors and tooth specific molecules were expressed both fetal and neonatal tissue. The tooth specific genes (enamelin, amelogenin, and DSPP) only expressed in neonate and adult stage. These expression patterns appeared same as primary fetal and neonatal cells. In this study we isolated primary cells from whole mandible of fetal and neonatal mice. And we investigated the characteristics of the primary cells and the profile of gene expressions, which are involved in epithelial-mesenchymal interactions during tooth development. Taken together, the primary dental cells in early passages or fetal and neonatal mandibles could be useful stem cell resources.

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotes stimulation of wound healing have been well known, there are few reports about molecular mechanism associated with cell cycle by LED irradiation. The purpose of present study was to examine the molecular event in cell cycle of LED irradiation on primary human gingival fibroblast(hGF) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635nm, and manufactured that energy density was 5mW/cm2 on sample surface. The hGF were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 8 and 24 hour after irradiation. To investigate the molecular mechanisms associated with cell cycle, growth phase was determined by flow cytometry and mRNA expression of cyclin A, cyclin B, cyclin D1, cyclin E, cdc2, PCNA, p18, p27, p21, and p53 were determined by real time RT-PCR. Flow cytometric analysis demonstrated the percentage of cells in the G1 and S phase were decreased, but the G2 phase increased, which showed cells irradiated by LED were transitioned from S to G2 phase. For mRNA expression, cyclin B, cdc2, PCNA and p53 were increased at 0 hour after irradiation, and most of cell cycle molecules were increased at 8 hour after irradiation. At 24 hour after irradiation, cyclin A, cyclin E, PCNA and p18 were increased. Taken together, LED irradiation induced proliferation of hGF cells through transition from S to G2 phase.

Stevia rebaudiana (Asteraceae), a perennial plant, has been used as a low-calorie sweetener and is being developed as a therapeutic agent for diabetes, hypertension, myocardial diseases, and microbial infections. Despite the common use of its leaves and stem, the bioavailability of the components present in S. rebaudiana flowers, when used as ingredients of cosmetics, has not been well investigated. Herein, we investigated the antioxidative and antimelanogenic effects of an aqueous extract of S. rebaudiana flowers (Stevia-F). Total flavonoid and phenolic content in Stevia-F were determined to be 8.64 ± 0.23 ㎎ of quercetin equivalents/100 g and 631.5 ± 2.01 ㎎ of gallic acid equivalents/100 g, respectively. The IC50 values of Stevia-F for reducing power, and 2,2-diphenyl-1-picryl-hydrazyl-hydrate radical, hydrogen peroxide, and nitric oxide scavenging activities were 5541.96, 131.39, 466.34, and 10.44 ㎍/mL, respectively. Stevia-F showed inhibitory effects on the tyrosinase (IC50 = 134.74 ㎍/mL) and α-glucosidase (IC50 = 114.81 ㎍/mL) activities. No significant cytotoxicity of Stevia-F was observed in B16F10 cells, treated with up to 100 ㎍/mL of the extract for 24 and 48 h (p > 0.05). Stevia-F (1–100 ㎍/mL) suppressed α-melanocyte stimulating hormone-induced melanin production in B16F10 cells (p < 0.05) and also inhibited the cellular tyrosinase activity (p < 0.05). Overall, our results show that Stevia-F possesses potential for inhibiting tyrosinase and α-glucosidase activities and has significant antioxidant capacity. The antimelanogenic potential of Stevia-F should extend the usage of S. rebaudiana flowers in the development of skinwhitening products.

본 연구의 목적은 토종다래의 용매별 추출물에 따른 약리활성에 대한 검증 및 효능 평가로서 토종다래의 항산화, 항염증에 대한 효과를 확인하였다. 염증 반응은 자극이 가해지면 histamine, serotonin, prostaglandin과 같은 혈관 활성물질에 의해 혈관 투과성이 증대되어 염증을 유발하고 cytokine, free radical, lysosomal enzyme 등 다양한 매개 인자가 관여한다. 자극에 의한 macrophage cell의 염증반응은 tumor necrosis factor-α(TNF-α), interleukin-6(IL-6), interleukin-1β(IL-1β)와 같은 pro-inflammatory cytokine의 발현이 유도되고, inducible nitric oxide synthase(i-NOS)와cyclooxygenase-2(COX-2)에 영향을 받는 유전자의 발현을 자극하게 되어 nitric oxide(NO) 등의 염증 인자가 생성된다. 이에 따라 토종다래 추출물의 항염증에 대한 연구를 위해 이에 영향을 주는 인자인 i-NOS, COX-2의 단백질 발현억제 작용을 확인하였다. 그 결과, HKE > HKA > HKW 순서로 높은 효능을 확인 할 수 있었다. 가장 효과가 좋은 HKE 처리군에서 다양한 염증성 인자의 mRNA 발현량을 확인하였다. 측정 결과, HKE(2,000 μg/mL)는 i–NOS, COX-2, IL-1β, IL-6, TNF-α mRNA 발현이 각각 93.2%, 27.9%, 96.4%, 89.4%, 73.9% 억제되는 효과를 확인할 수 있었다. 또한, HKE의 nuclear factor kappa B(NF-κB) 단백질 발현에 농도의존적으로 유의미한 결과를 확인하였으며, 이에 토종다래의 항염증효과는 LPS에 의한 TLR4의 자극에서 NF-κB 경로의 완화로 나타는 것임을 검증하였다. 결론적으로 토종다래는 70% ethanol 추출물(HKE)의 항염증 효과가 가장 높았으며, HKE는 대식세포에서 NF-κB 염증관련 경로의 억제로 세포 내 mRNA 및 단백질 수준에서의 염증인자들의 생성을 저해하여 항염증 효과가 명백히 확인되었다. 향후 본 연구팀은 토종다래의 항염증과 관련된 유효성분의 분리정제 및 구조분석을 진행할 예정이다.

Angelica tenuissima, also known as Ligusticum tenuissimum, is classified as a food-related plant and has been used as traditional medicines treating headache and anemia in Asia. However, its anti-melanogenic effect has not been reported in detail. When the extract of Angelica tenuissima (ATE) was prepared by the extraction with 70% EtOH at 80°C (final yield = 22%), the contents of decursin and Z-ligustilide in ATE were determined 0.06% and 8.43%, respectively. Total flavonoid and phenolic content in mg ATE were 5.52±0.07 ㎍ quercetin equivalents and 237.27±13.24 ㎍ gallic acid equivalents, respectively. Antioxidant capacity of ATE determined by DPPH and ABTS assay was increased with a dose dependent manner up to 1000 ㎍/㎖. The amount of melanin synthesis followed by α-melanocyte stimulating hormone on B16F10 cells were significantly reduced in the presence of ATE (250 to 1000 ㎍/㎖, p<0.05). ATE (125 to 1000 ㎍/㎖, p<0.05) suppressed the tyrosinase activity but did not show any significant effect on α-glucosidase activity at the same condition. Taken together, ATE possesses tyrosinase inhibitory potential with significant antioxidant capacities. These effects of ATE might be involved in suppression of melanin synthesis, at least, in B16F10 cells. The anti-melanogenic potential of ATE will provide an insight into developing a new skin whitening product.

The aim of this study is to investigate the antioxidant and intracellular anti-inflammatory efficacy of blueberry leaf extracted with hot water (BLW), 70% ethanol (BLE), and 70% acetone (BLA) in RAW 264.7 macrophages. In order to evaluate the anti-inflammatory effect of blueberry leaf extracts, RAW 264.7 macrophages were stimulated with lipopolysaccharide (LPS) to induce the production of inflammation-related factors, which were measure by Western blotting and real-time PCR methods. i-NOS, COX-2 protein, and mRNA expression showed concentration-dependent decrease. The decreases in the mRNA expression levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and prostaglandin E2 (PGE2) were concentration-dependent. Further, the antioxidant effects of blueberry leaf on total polyphenol contents, electron donating ability and ABTS+ radical scavenging activity were evaluated. The total polyphenol contents of BLW, BLE, and BLA were 217.04±2.98, 156.72±3.90, and 182.88±3.02 mg TAE/g, respectively, while the electron donating abilities at 1,000 μg/mL of BLW, BLE, and BLA were 81.7, 79.6, and 79.3%, respectively. The ABTS+ radical scavenging activity was fond to be concentration dependent. The nitric oxide (NO) production inhibition activities at 50 μg/mL of BLW, BLE, and BLA were 35.1, 42.4 and 42.7%, respectively. In conclusion, the antioxidant and anti-inflammatory test results indicate that blueberry leaf extracts (BLW, BLE, and BLA) can be used as potential anti-inflammatory agents.

EP was obtained through 20% ethanol extraction of Pueraria lobata root, and the fermented form of EP, FEP, was prepared from the EP after incubating with Lactobacillus rhamnosus vitaP1. There was no significant toxicity by EP and FEP up to 1000 ㎍/㎖ in NIH-3T3, HaCaT, and B16F10 cells. In addition to antioxidant potentials of EP and FEP determined by DPPH and ABST assays, we confirmed increase of procollagen type I and elastin synthesis by supplementation of the EP and FEP at the concentration of 50 ㎍/㎖ using ELISA kits. The protein expression levels of matrix metalloprotease (MMP)-1, -3, and -9, those are involved in the degradation of collagen or other skin matrix proteins, were remarkably suppressed while their inhibitory protein metallopeptidase inhibitor 1 (TIMP-1) was greatly up-regulated by supplementation of the EP and FEP at a concentration of 50 ㎍/㎖. Taken together, both EP and FEP supplementation could be involved in the suppression of the skin wrinkle formation through inhibiting degradation of collagen and stimulating the synthesis of collagen and elastin. The results showed that the anti-wrinkle potential of the EP and FEP will be a promising candidate for developing cosmeceutical compounds or products.

Backgoound : This study was conducted to serve as a basis for the selection of superior lines by analyzing the content of antioxidant component and antioxidant activity in Schisandra chinensis Collections Methods and Results : In order to examine antioxidant component and antioxidant activity, 154 species of Schisandra chinensis from whole country were used. Antioxidant component was investigated by total flavonoid content and total phenolic content and antioxidant activity was evaluated by DPPH radical scavenging activity and ABTS radical scavenging activity. As a result, the total amounts of flavonoids was highest in SC-20 with 5.03 ㎎/g. However the content of polyphenols showed highest in SC-22 with 2.76 ㎎/g. In addition antioxidant activity results were also relatively high in SC-22. The IC50 value was 548 ㎍/㎖ in DPPH radical scavenging and 640 ㎍/㎖. in ABTS+ radical scavenging. Conclusion : As demand for high income crop has increased, new cultivar breeding is required to produce high quality Schisandra chinensis From this study, analyses of antioxidant component and antioxidant activity in collection can be used for new Schisandra chinensis breeding. Especially SC-22 can be superior lines with high antioxidant component and antioxidant activity.

Background : Undulatum Rhubarb, commonly produced in domestic, is rhizome of Rheum undulatum L. that belongs to the family Polygonaceae. It also can be used as a substitute of R. palmatum L., R. tanguticum Maximowicz ex Balf., and R. officinale Baillon which completely depend on import system. However, there should be clear clarification among Undulatum Rhubarb and Rhubarb, because Undulatum Rhubarb contains rhaponticin as marker compound that is not indicated at Rhubarb. Some of the recently imported Undulatum Rhubarbs have been found to be Rhubarb. Also, it is known that only Undulatum Rhubarb is cultivated at domestic environment. But some plant origins of Rhubarb are grown in Korea, too. Further study are needed to clarify clear origin between Undulatum Rhubarb and Rhubarb. Thus, we collected some domestically cultivated samples and identified them. Methods and Reseults : Rheum undulatum L., Rhubarb, Rheum tanguticum Maximowicz ex Balf. which were cultivated in Gangwondo Agricultural Research and Extension Services in Cheorwon were collected and anayzed the DNA sequences. We also compared DNA sequences in Rhubarb collected from England and R. rhabarbarum L., R. undulatum L., and R. franzenbachii on NCBI. As a result, two kinds of rhubarb cultivated in the test plantation were identified as R. rhabarbarum and R. officinale. In addition, R. undulatum (plant origin of Undulatum Rhubarb) was identified as Rhubarb (Rheum rhabarbarum) in England with 99 - 100% identical in nuclear ITS gene region. Conclusion : R. undulatum, plant origin of Undulatum Rhubarb, is reported as synonym of R. rhabarbarum, R. franzenbachii. Rheum speices which are cultivated as tester in Gangwondo Agricultural Research and Extension Services in Cheorwon are estimated as R. undulatum and R. officinale. Therefore, not only Undulatum Rhubarb but Rhubarb could be grown in Korea.

Background : In previous studies, adlay seeds showed a prevalence of diversified fungal flora with the predominant fungal genera being Fusarium (45.6%) and In vitro test showed that fungal toxins like Fumonisin and Zearalenone were produced by Fusarium fujikuroi, F. asiaticum and F. graminearum. Because of this we performed experiments to selecting disinfective chemicals for controlling the Fusarium contamination in the adlay seed. Methods and Results : We carried out the chemical efficacy test such as seed disinfectants selection test appling before planting and pesticides selection test using in the earing season. In the present study, eleven different commercially available seed disinfectant were applied to the adlay seeds. Among 11 seed disinfectants, Hexaconazole+Prochloraz emulsifiable concentrate (EC) and Prochloraz emulsifiable concentrate (EC) had control value of 80% or above against Fusarium species tested. In the pesticides selection test, seven different commercially available pesticides for Fusarium blight (Scab) control were applied and we observed that Metconazole suspension concentrate(SC) strongly inhibited the mycelial growth of 10 Fusarium species all. Conclusion : From the above results, we selected Hexaconazole+Prochloraz EC and Prochloraz EC as a seed disinfectants and Metconazole SC as a pesticide using in the earing season for Scab control.

Background : This experiment was conducted to select GAP applying seed disinfectants in Astragalus membranaceus and Platycodon grandiflorum. Methods and Results : We carried out the chemical efficacy and injury test. For the efficacy test, we investigated fungal detection rate by seed disinfectants and for the chemical injury, we investigated germination rate and emergence rate by seed disinfectants in reference amount and fold amount. These experiments carried out two times. The results obtained are as follows. In the experiment for seed disinfectants selection of Astragalus membranaceus, all tested chemicals such as Tebuconazole emulsifiable concentrate(EC), Thiophanate-methyl + Triflumizole wettable powder(WP), Prochloraz copper chloride complex+Tebuconazole suspension concentrate(SC), Prochloraz emulsifiable concentrate(EC), Fludioxonil wetting liquid(WL) and Hexaconazol+Prochloraz emulsifiable concentrate(EC) had control value of 80% or above against seed contaminated fungi. However two chemicals such as Tebuconazole EC and Prochloraz copper chloride complex+Tebuconazole SC and two chemicals such as Prochloraz EC and Hexaconazol+Prochloraz EC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. In the case of seed disinfectants selection of Platycodon grandiflorum, Prochloraz copper chloride complex+Tebuconazole SC, Prochloraz EC and Hexaconazol+Prochloraz EC had control value of above 80% against seed contaminated fungi except Thiophanate-methyl+Triflumizole WP and Fludioxonil WL. However Hexaconazol+Prochloraz EC and Prochloraz copper chloride complex+Tebuconazole SC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. Conclusion : From the above results, we finally selected three items of seed disinfectants including Thiophanate-methyl+Triflumizole WP and Fludioxonil WL in Astragalus membranaceus and Prochloraz emulsion in Platycodon grandiflorum.

Background : The objective of this study was to investigate antioxidant activities, inhibitory activities against heme induced colonic epithelial cell proliferations, anti-inflammatory activities and anthocyanin profiles in the anthocyanin rich fraction (ARFAM) from fruits of Aronia melanocarpa, where these are considered functional substances and available food coloring agents in Korea. Methods and Results : Anthocyanins were identified by reversed-phase C18 column chromatography and HPLC-DAD-ESI/MS analysis. To compare the antioxidative and anti-inflammatory capacity of Aronia melanocarpa berries, recognized for their high content of anthocyanins, isolation method was developed to obtain high-purity anthocyanins in the extract. Anthocyanin-rich fractions (ARFAM) enriched in anthocyanins were found to be potent strong inhibitory activity towards heme induced colonic epithelial cell proliferations are associated with an increased risk of colon cancer than acidic ethanol extract (AME). The immunomodulation properties were assessed in growth of both human B and T cells, its cytokines secretion such as IL-6 (interleukin-6) and TNF-α (tumor necrosis factor-alpha). AME enhanced interleukin-6 and reduced tumor necrosis factor-a production, whereas ARFAM only had a effect in increasing of IL-6 expression. Conclusion : These results demonstrated that there was no major relationship between the antioxidative and immunomodulation capacities of AME and ARFAM.