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Oxidative Inactivation of Peroxiredoxin Isoforms by H2O2 in Pulmonary Epithelial, Macrophage, and other Cell Lines with their Subsequent Regeneration (폐포상피세포, 대식세포를 비롯한 각종 세포주에서 H2O2에 의한 Peroxiredoxin 동위효소들의 산화에 따른 불활성화와 재생)

  • Oh, Yoon Jung;Kim, Young Sun;Choi, Young In;Shin, Seung Soo;Park, Joo Hun;Choi, Young Hwa;Park, Kwang Joo;Park, Rae Woong;Hwang, Sung Chul
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.31-42
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    • 2005
  • Background : Peroxiredoxins (Prxs) are a relatively newly recognized, novel family of peroxidases that reduce $H_2O_2$ and alkylhydroperoxide into water and alcohol, respectively. There are 6 known isoforms of Prxs present in human cells. Normally, Prxs exist in a head-to-tail homodimeric state in a reduced form. However, in the presence of excess $H_2O_2$, it can be oxidized on its catalytically active cysteine site into inactive oxidized forms. This study surveyed the types of the Prx isoforms present in the pulmonary epithelial, macrophage, endothelial, and other cell lines and observed their response to oxidative stress. Methods : This study examined the effect of exogenous, excess $H_2O_2$ on the Prxs of established cell lines originating from the pulmonary epithelium, macrophages, and other cell lines, which are known to be exposed to high oxygen partial pressures or are believed to be subject to frequent oxidative stress, using non-reducing SDS polyacrylamide electrophoresis (PAGE) and 2 dimensional electrophoresis. Result : The addition of excess $H_2O_2$ to the culture media of the various cell-lines caused the immediate inactivation of Prxs, as evidenced by their inability to form dimers by a disulfide cross linkage. This was detected as a subsequent shift to its monomeric forms on the non-reducing SDS PAGE. These findings were further confirmed by 2 dimensional electrophoresis and immunoblot analysis by a shift toward a more acidic isoelectric point (pI). However, the subsequent reappearance of the dimeric Prxs with a comparable, corresponding decrease in the monomeric bands was noted on the non-reducing SDS PAGE as early as 30 minutes after the $H_2O_2$ treatment suggesting regeneration after oxidation. The regenerated dimers can again be converted to the inactivated form by a repeated $H_2O_2$ treatment, indicating that the protein is still catalytically active. The recovery of Prxs to the original dimeric state was not inhibited by a pre-treatment with cycloheximide, nor by a pretreatment with inhibitors of protein synthesis, which suggests that the reappearance of dimers occurs via a regeneration process rather than via the de novo synthesis of the active protein. Conclusion : The cells, in general, appeared to be equipped with an established system for regenerating inactivated Prxs, and this system may function as a molecular "on-off switch" in various oxidative signal transduction processes. The same mechanisms might applicable other proteins associated with signal transduction where the active catalytic site cysteines exist.

Phytochrome Signal Transduction Regulates Anthocyanin Biosynthesis in Cell Suspension Cultures of Vitis vinifera (포도 세포현탁배양계에서 Phytochrome 신호전달에 의해 조절되는 안토시아닌 생합성)

  • Choi, Kwan-Sam;Kim, Sun-Kyung;In, Jun-Gyo;Shin, Dong-Ho
    • Journal of Plant Biotechnology
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    • v.31 no.3
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    • pp.239-248
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    • 2004
  • This experiment was carried out to confirm that phytochrome regulates anthocyanin bio-synthesis during cell suspension culture system of grape or not. In suspension culture of grape, maximum accumulation of anthocyanin was observed at the stationary phase under continuous white light condition. From mono-chromatic light interruption for 24h at the 4th or 7th day on the suspension cultured cells, the anthocyanin accumulation was highly enhanced at the light interruption at 7th day than 4th day under all monochromatic light treatment. However, the cell growth patterns were not affected by any light treatment. In the darkness, the anthocyanin synthesis was very low but remarkably increased by blue light or red light irradiation. However, the increase of anthocyanin accumulation by blue or red light was suppressed by far-red light in the suspension cells of grape. This suppression by far-red light on the anthocyanin synthesis also observed on the cells treated red or far-red light alternatively. These results implied that phytochrome regulation system may be involved in the anthocyanin biosynthesis of the suspension grape cells. By RNA expression analysis, chalcone synthase (CHS) gene was expressed highly by blue and red light but low by far-red light. The synergistic increase of CHS gene expression was also observed at the treatment of blue light followed by red for 24h. This result may explain the increase of anthocyanin accumulation in B/R treatment. Although the expression of phytochrome gene (PHYA or PHYB) was not highly increased by all light treatment (blue, red, and far-red light) the expression of both PHYA gene and PHYB gene was increased a little in cells treated red or far-red light. In grape suspension cells, the red light enhanced the anthocyanin synthesis, whereas the far-red light was suppressed. Although it was not confirmed whether or not phytochrome gene is activated in anthocyanin accumulating grape cells, we believed that anthocyanin biosynthesis in grape cells may be regulated under phytochrome signal transduction system.

2DEG Transport Analysis in AlGaAs/GaAs Interface by MONTE-CARLO Method (MONTE-CARLO 방법에 의한 AlGaAs/GaAs 계면의 전자 전달특성 분석)

  • Nam, Seung-Hun;Jung, Hak-Ki;Kim, Bong-Ryul
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.26 no.2
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    • pp.94-101
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    • 1989
  • Transport properties of 2DEG at AlGaAs/GaAs interface such as average electron energy, flight distance, each valley occupancy ratio, average electron velocity for various fields are investigated by MONTE-CARLO method. As the electric field increases, more electrons transit drastically from (000) valley to (000) upper valley. This phenomenon shows the nonstationary effect such as velocity overshoot. The duration of the transient decreases from about 1.4 psec for electric field E = 7KV/cm to about 0.7 psec for 12KV/cm. The average electron velocity during transient transport in 2DEG is about 8 times the steady-state velocity for E = 12KV/cm at room temperature. In comparison with bulk GaAs the peak velocity in the 2DEG is higher than that in even pure bulk GaAs at electric field E = 7 KV/cm. On the basis of the fact that the electrons in the 2DEG have larger peak velocity and shorter transient time of velocity than those in the bulk GaAs, it is suggested that the device with 2DEG may obtain higher mobility than that with bulk GaAs.

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A review of the contents about childhood health care in middle & high school textbooks (중·고등학교 교과서에 실린 소아 관련 정보의 조사)

  • Kim, Jung Hun;Park, Sung Won;Shin, Son Moon;Sung, In Kyung;Park, Mi Jung;Chung, Yoo Mi;Ha, Jeong Hun
    • Clinical and Experimental Pediatrics
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    • v.50 no.4
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    • pp.340-347
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    • 2007
  • Purpose : The purpose of this study is to investigate the material in middle school and high school textbooks covering child care and children's health for medically inaccurate content and to make sure that the correct information is given to students. Methods : We have examined 47 kinds of textbooks on the subjects 'Home economics' and 'Technology and home economics' published in 2007 to find out how much importance is placed on children's health and childcare and to search for incorrect definitions, inaccurate facts or insufficient explanations. We have also analyzed the credentials of the writers of these textbooks. Results : Textbooks of middle school cover psychological and physical changes during adolescence and nutrition, and those of high school cover marriage and childcare. These materials covering health made up 27.5%, 11.7% of middle & high school texts. Age definitions of neonate and infant were incorrect. Inaccurate facts were mostly about fontanelles, primitive reflexes, breastfeeding and weaning. There also were wrong informations on the umbilical cord care and developmental milestones during infancy. The childhood immunization schedules and the growth curves were not up to date. Most of the authors major in home economics and the others were school teachers. There was no evidence of any review by a medical society or an expert. Conclusions : When writing about children's health and childcare in textbooks, a board-certified pediatrician or The Korean Pediatric Society should be consulted to provide accurate medical information to middle and high school students.

The Market Segmentation of Coffee Shops and the Difference Analysis of Consumer Behavior: A Case based on Caffe Bene (커피전문점의 시장세분화와 소비자행동 차이 분석 : 카페베네 사례를 중심으로)

  • Yu, Jong-Pil;Yoon, Nam-Soo
    • Journal of Distribution Science
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    • v.9 no.4
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    • pp.5-13
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    • 2011
  • This study provides analysis of the effectiveness of domestic marketing strategies of the Korean coffee shop "Caffe Bene". It bases its evaluation on statistical outputs of 'choice attributes,' "market segmentation," demographic characteristics," and "satisfaction differences." The results are summarized in four points. First, five choice attributes were extracted from factor analysis: price, atmosphere, comfort, taste, and location; these are related to coffee shop selection behavior. Based on these five factors, cluster analysis was conducted, with statistical results classifying customers into three major groups: atmosphere oriented; comfort oriented; and taste oriented. Second, discriminant analysis tested cluster analysis and showed two discriminant functions: location and atmosphere. Third, cross-tabulation analysis based on demographic characteristics showed distinctive demographic characteristics within the three groups. Atmosphere oriented group, early-20s, as women of all ages was found to be 'walking down the street 'and 'through acquaintances' in many cases, as the cognitive path, and mostly found the store through 'outdoor advertising', and 'introduction'. Comfort oriented group was mainly women who are students in their early twenties or professionals, and appeared as a group to be very loyal because of high recommendation to other customers compared to other groups. Taste oriented group, unlike the other group, was mainly late-20s' college graduates, and was confirmed, as low loyalty, with lower recommendation activity. Fourth, to analyze satisfaction differences, one-way ANOVA was conducted. It shows that groups which show high satisfaction in the five main factors also show high menu satisfaction and high overall satisfaction. This results show that segmented marketing strategies are necessary because customers are considering price, atmosphere, comfort, taste, location when they choose coffee shop and demographics show different attributes based on segmented groups. For example, atmosphere oriented group is satisfied with shop interior and comfort while dissatisfied with price because most of the customers in this group are early 20s and do not have great financial capability. Thus, price discounting marketing strategies based on individual situations through CRM system is critical. Comfort oriented group shows high satisfaction level about location and shop comfort. Also, in this group, there are many early 20s female customers, students, and self-employed people. This group customers show high word of mouth tendency, hence providing positive brand image to the customers would be important. In case of taste oriented group, while the scores of taste and location are high, word of mouth score is low. This group is mainly composed of educated and professional many late 20s customers, therefore, menu differentiation, increasing quality of coffee taste and price discrimination is critical to increase customers' satisfaction. However, it is hard to generalize the results of study to other coffee shop brand, because this study have researched only one domestic coffee shop, Caffe Bene. Thus if future study expand the scope of locations, brands, and occupations, the results of the study would provide more generalizable results. Finally, research of customer satisfactions of menu, trust, loyalty, and switching cost would be critical in the future study.

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Cytoprotective Effects of Phaeophyta Extracts from the Coast of Jeju Island in HT-22 Mouse Neuronal Cells (제주 연안 갈조류 추출물의 신경세포 보호효과)

  • Shin, Dong-Bum;Han, Eun-Hye;Park, Sung-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.224-230
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    • 2014
  • Marine algae have long been recognized as a health and beauty food, based on its anti-tumor, anti-inflammatory and anti-obesity activities. In this study, methanol extracts were prepared from 10 different phaeophyta, after which DPPH radical scavenging and cytoprotective activities of HT-22 cells against ${\beta}$-amyloid protein ($A{\beta}$), which has neurotoxic effects, were investigated. In DPPH experiments, Ecklonia cava and Ishige okamurai showed strong ROS scavenging activities, whereas eight other phaeophyta including Petalonia binghamiae (P. bin) showed weak ROS scavenging activities. To validate the cytoprotective effects of 10 different phaeophyta in $A{\beta}$-induced HT-22 cells, protein expression levels of APP, BACE1, iNOS, phosphorylated ERK1/2, phosphorylated p38 and phosphorylated JNK1/2 were determined along with MTT assay. In the MTT assay, P. bin showed the best effective cytoprotective activity at a concentrations of $25{\mu}g/mL$, whereas Sargassum confusum, Colpomenia sinuosa, Myelophycus simplex, and Sargassum hemiphyllum showed potential. Determination of protein expression levels related to $A{\beta}$-induced neurotoxicity in the five selected phaeophyta showed that P. bin inhibited BACE1 and iNOS expression in $A{\beta}$-induced HT-22 cells. These results indicate that the cytoprotective effects of P. bin are mediated by suppressing the pathways involving $A{\beta}$-induced ERK and p38 activation.

Global DNA Methylation Patterns and Gene Expression Associated with Obesity-Susceptibility in Offspring of Pregnant Sprague-Dawley Rats Exposed to BDE-47 and BDE-209 (임신 중 BDE-47 및 BDE-209에 노출된 어미와 새끼 Sprague-Dawley 랫드의 Global DNA 메틸화 양상과 비만 감수성과 연관된 유전자 발현)

  • Park, Byeong-Min;Yoon, Ok-Jin;Lee, Do-Hoon
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.1
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    • pp.28-39
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    • 2017
  • Persistent organic pollutants (POPs) can affect epigenetic mechanisms and obesity development. Polybrominated diphenyl ethers (PBDEs)-widely used to make flames-are one of the important POPs. Prenatal exposure to endocrine disrupting chemicals (EDCs), such as POPs, may affect global DNA methylation in long interspersed nuclear elements (LINE-1), increasing the risk of obesity later in life. Therefore, pregnant Sprague-Dawley (SD) rats were used to elucidate whether BDE-47 and BDE-209 transferred through placenta and breast milk cause epigenetic changes in LINE-1 and increase genetic susceptibility to obesity as obesogen during the developmental periods. Global DNA methylation in LINE-1 and gene expression related to obesity were measured in dams and offspring, using a methylation-sensitive high resolution melting analysis (MS-HRM) and direct bisulfite sequencing and quantitative real time polymerase chain reaction (qPCR), respectively. The results of MS-HRM showed global DNA hypomethylation patterns in LINE-1 of exposed offspring (2 of total 4) at PND 4, but bisulfite sequencing showed no difference in both the exposed and non-exposed groups. Gene expression in dams related to ${\beta}$-oxidation pathway and those related to adipokines showed different patterns between the two groups. On the contrary, gene expressions of offspring showed a similar pattern. Gene expressions related to ${\beta}$-oxidation pathway and obesity were significantly increased when compared with 'at birth', but not $PPAR-{\alpha}$. In conclusion, this study demonstrated the possibility that co-exposure to BDE-47 and BDE-209-via the placenta and breast milk-may affect epigenetic changes and modulate gene expression levels related to obesity.

Studies on the Transovarial Transmission of Cytoplasmic Polyhedrosis Virus with Reference to the Phenomena of Induction, Interference and Virulence Enhancement in the Silkworm, Bombyx mori (가잠 세포질다각체 바이러스의 유발, 간섭 및 병원성 증진현상에 의한 경난전달에 관한 연구)

  • 임종성
    • Journal of Sericultural and Entomological Science
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    • v.16 no.2
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    • pp.55-75
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    • 1974
  • Transovarial transmission of cytoplamic polyhedrosis virus in the silkworm was studied by observing the phenomena of induction, interference and virulence enhancement in the larvae from moths inoculated with hexagonal Polyhedra of cytoplasmic polythedrosis virus. The experimental results obtained are as followings. 1. The effect of inoculation with tetragonal polyhedra of cytoplasmic polyhedrosis virus on the rate of infective induction with hexagonal polyhedron virus and with hexagonal plus tetragonal polyhedron viruses in the larvae from moths infected with hexagonal polyhedron virus was studied. Infection rate was higher by 40 to 60 percent in the larvae from infected female group than in tile larvae from noninfected female group. 2. In the studies of the effect of formalin-feeding on the induction of infection with hexagonal polyhedron virus, infection rate was higher by 40 percent in the larvae from infected female group than in the larvae from noninfected female group. However, there was no significant difference in the infection rates between the two formalin-concentration groups. 3. The effect of cold treatment on the induction of infection with hexagonal polyhedron virus was studied. Infection rate was higher by 50 percent in the larvae from infected female group than ill the larvae from noninfected female group. No difference was found in the infection rates of the two treatment groups of 12 and 48 hours. 4. The phenomena of induction and interference were studied by observing rate of infection with hexagonal polyhedron virus induced by the inoculation with tetragonal polyhedron virus. The degree of interference of primary hexagonal polyhedron virus by secondary tetragonal Ployhedron virus was increased as the dosage of secondary virus was increased. At the concentration of 1${\times}$10$\^$8/m1 of the secondary virus, the degree of interference was similar to. that of control group. On the other hand, infection with tetragonal polyhedron virus at low concentration was interfered by the primary virus. At the concentration of 1N10f/m1 of tetragonal polyhedron virus, however, the rate of infection with tetragonal polyhedron virus was increased sharply, which is still lower by 30 percent than that of control group. 5. In the studies of induction and virulence enhancement, based on the 1ate of mixed infection with hexagonal and tetragonal polyhedron viruses, the highest difference of infection rate between experimental group and control group exceeded 40 percent when the concentration of tetragonal polyhedron virus was 1${\times}$10$\^$7/m1. However, the infection rate of control group was not affected by concentrations of tetragonal polyhedron virus.

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A Study on the Direction of Restoration for Baegun-dong Garden in Gangjin through a Diachronic Interpretation of the Prototype Scenery (통시적 원형경관 해석을 통해 본 강진 백운동 원림의 복원 방향)

  • Ha, Hye-Kyung;So, Hyun-Su
    • Journal of the Korean Institute of Landscape Architecture
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    • v.45 no.3
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    • pp.116-128
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    • 2017
  • In this study, the original scenery of Baegun-dong Garden in Gangjin was compared with that of the reconstructed site to suggest directions for restoration in the future. The diachronic prototype scenery of the Baegun-dong Garden was derived by integrating the elements of the original and contemporary prototype sceneries from the old poetry series. The old poetry series is comprised of "Baegun-dong 8Yeong(詠)" by Lee Damro(李聃老), "Baegun-dong 10Su(首)" by Song Ikhwi(宋翼輝), "Baegun-dong 12Gyeong (景)" by Jeong Yakyong(丁若鏞), and "Baegun-dong 14Gyeong" by Lee Siheon (李時憲). Guidance for directions for future restoration was suggested through a comparative analysis between the diachronic prototype scenery of Baegun-dong Garden and the current scenery of the restored site. The research result is as follows. First, the prototype scenery of Baegun-dong Garden was a harmonic view composed of the camellia forest road, 'Baegun-dong' the letter carved on a rock, a cliff 'Changhabyeok(蒼霞壁)' the red letter written by Jeong Yakyong, paulownia trees, waterfalls and a maple forest. Outside the Byeolseo(別墅), there were a red plum blossom forest, a pine forest, and a bamboo forest, as well as the view of Okpanbong(玉版峯) from Jeongseondae(停仙臺). In the yard, there were Chwimiseonbang(翠微禪房), Jug-gak(竹閣), Pyeongsang(榻), a lotus Bangji(方池), Gogsugeo(曲水渠) and Hwagye(花階). Cranes were even raised in the yard. Second, comparing the diachronic prototype scenery of Baegun-dong Garden with its restored site, following a restoration guide is suggested. There should be more water flow in Baegun-dong Valley and Jug-gak should be restored. 'Changhabyeok' the red letter should be carved on the stone wall. Trees should be controlled for a better view of Okpanbong from Jeongseondae. More trees and plants, such as red plum blossom, pine tree, paulownia trees, azaleas(映山紅), chrysanthemum, orchid, daffodil, and lotus should be planted in the yard. A system of integrated guide boards is also required for effective information transmission for visitors.

Antioxidant and Cytoprotective Effects of Socheongja and Socheong 2, Korean Black Seed Coat Soybean Varieties, against Hydrogen Peroxide-induced Oxidative Damage in HaCaT Human Skin Keratinocytes (HaCaT 인간 피부 각질세포에서 과산화수소 유도 산화 손상에 대한 소청자 및 소총2호의 항산화 및 세포보호 효능)

  • Choi, Eun Ok;Kwon, Da Hye;Hwang, Hye-Jin;Kim, Kook Jin;Lee, Dong Hee;Choi, Yung Hyun
    • Journal of Life Science
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    • v.28 no.4
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    • pp.454-464
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    • 2018
  • Black soybeans are used as food sources as well as for traditional medicines because they contain an abundance of natural phenolic compounds. In this study, total phenolic contents (TPCs) of Korean black seed coat soybean varieties Socheongja (SCJ), Socheong 2 (SC2) and Cheongja 2 (CJ2) as well as their antioxidant capacities were investigated. Among them, TPCs were abundantly present in the order of CJ2$H_2O_2$-stimulated HaCaT human keratinocytes. Our results revealed that treatment with SCJ and SC2 prior to $H_2O_2$ exposure significantly increases the viability of HaCaT cells, indicating that the exposure of HaCaT cells to SCJ and SC2 conferred a protective effect against oxidative stress. SCJ and SC2 also effectively inhibited $H_2O_2$-induced apoptotic cell death through the blocking of mitochondrial dysfunction. SCJ and SC2 also attenuated the phosphorylation of Histone H2AX. Furthermore, they effectively induced the levels of thioredoxin reductase (TrxR) 1, a potent antioxidant enzyme, which is associated with the induction of nuclear transcription factor erythroid-2-like factor 2 (Nrf2); however, the protective effects of SCJ and SC2 were significantly reversed by Auranofin, a TrxR inhibitor. These results indicate that they have protective activity through the blocking of cellular damage related to oxidative stress via the Nrf2 signaling pathway. In conclusion, our study indicated that SCJ and SC2 might potentially serve as novel agents for the treatment and prevention of skin disorders caused by oxidative stress.