• Title/Summary/Keyword: Bio-marker

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Ginsenoside Rb2 suppresses cellular senescence of human dermal fibroblasts by inducing autophagy

  • Kyeong Eun Yang;Soo-Bin Nam;Minsu Jang;Junsoo Park;Ga-Eun Lee;Yong-Yeon Cho;Byeong-Churl Jang;Cheol-Jung Lee;Jong-Soon Choi
    • Journal of Ginseng Research
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    • v.47 no.2
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    • pp.337-346
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    • 2023
  • Background: Ginsenoside Rb2, a major active component of Panax ginseng, has various physiological activities, including anticancer and anti-inflammatory effects. However, the mechanisms underlying the rejuvenation effect of Rb2 in human skin cells have not been elucidated. Methods: We performed a senescence-associated β-galactosidase staining assay to confirm cellular senescence in human dermal fibroblasts (HDFs). The regulatory effects of Rb2 on autophagy were evaluated by analyzing the expression of autophagy marker proteins, such as microtubule-associated protein 1A/1B-light chain (LC) 3 and p62, using immunoblotting. Autophagosome and autolysosome formation was monitored using transmission electron microscopy. Autophagic flux was analyzed using tandem-labeled GFP-RFP-LC3, and lysosomal function was assessed with Lysotracker. We performed RNA sequencing to identify potential target genes related to HDF rejuvenation mediated by Rb2. To verify the functions of the target genes, we silenced them using shRNAs. Results: Rb2 decreased β-galactosidase activity and altered the expression of cell cycle regulatory proteins in senescent HDFs. Rb2 markedly induced the conversion of LC3-I to LC3-II and LC3 puncta. Moreover, Rb2 increased lysosomal function and red puncta in tandem-labeled GFP-RFP-LC3, which indicate that Rb2 promoted autophagic flux. RNA sequencing data showed that the expression of DNA damage-regulated autophagy modulator 2 (DRAM2) was induced by Rb2. In autophagy signaling, Rb2 activated the AMPK-ULK1 pathway and inactivated mTOR. DRAM2 knockdown inhibited autophagy and Rb2-restored cellular senescence. Conclusion: Rb2 reverses cellular senescence by activating autophagy via the AMPK-mTOR pathway and induction of DRAM2, suggesting that Rb2 might have potential value as an antiaging agent.

The Gene Pyramiding Effect of qBK1 and qFfR1 Genes of Major Resistance Genes in Bakanae Disease

  • Sais-Beul Lee;Sumin Jo;Jun-Hyun Cho;Nkulu Rolly Kabange;Ji-Yoon Lee;Yeongho Kwon;Ju-Won Kang;Dongjin Shin;Jong-Hee Lee;You-Cheon Song;Jong-Min Ko;Kyung-Min Kim;Dong-Soo Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2020.12a
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    • pp.65-65
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    • 2020
  • Bakanae disease, caused by Gibberella fujikuroi, is one of the most devastating diseases threatening rice production in Korea. In recent years, the incidence of bakanae disease became alarming due to the mechanical transplanting practice where the spread of bakanae can be amplified during accelerating seeds growth, due to the use of seeding boxes. The development of resistant rice cultivars could be the primary and effective method for controlling bakanae disease. However, the effects of individual resistance genes are relatively small. Therefore, pyramiding of bakane R genes in rice breeding is a promising strategy having a high potential to mitigate the advert effects of bakanae disease. This study employed a gene pyramiding approach to develop bakanae disease resistant rice lines carrying qBK1, qFfR1 introduced from rice line MY299BK and cv. Nampyeong, respectively. The MY299BK carries qBK1 introduced from cv. Shingwang, which was found to have a high resistance compare to Nampyeong. In addition, the pyramiding effect of the qBK1 and qFfR1 resistance genes were investigated, and the presence or absence of these genes helped us investigate their interaction through bioassay method and MAS. Furthermore, the distribution of resistance in the population showed a biased distribution toward resistance in the F6:7 populutions. However, we could not confirm the accumulation effect of the resistance gene, but the difference between the two genes by the SN2 marker was confirmed. Therefore, the qBK1 gene harbored by MY299BK appears to be different from the qFfR1 carried by Nampyeong, suspected to possess a different bakanae disease resistant gene different from those found in MY299BK and Nampyeong.

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Establishment of Reference Intervals of Osteocalcin according to Age in Women for Health Promotion Center (건강검진이 의뢰된 여성의 연령에 따른 Osteocalcin의 참고범위 설정)

  • Kang, Ji-Soon;Yoo, Byoung-Joo;Oh, Jung-Eun;Kim, Geon-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.13 no.1
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    • pp.104-111
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    • 2009
  • Purpose: Osteocalcin is also known as the bone gamma-carboxyglutamic acid (Gla) protein (BGP), is noncollagenous bone protein synthesized by osteoblasts. Serum concentrations of Osteocalcin have been used as a biochemical marker of bone turnover. The reference intervals of Osteocalcin is categorized by kit corporation according to the age. However, each laboratory should establish its own reference intervals. In this study, the variation in the serum Osteocalcin level were used to find actual standard age-specific Osteocalcin reference intervals. Materials and Methods: We have selected 864 healthy females aged 20~80 years who visited a health promotion center between Aug. 2007 and Sep. 2008. The Osteocalcin IRMA Kit (OSTEO-RIACT, CIS Bio international, Gif-sur-Yvette, France) was used for the quantification. Each results were analyzed with the SPSS 12.0 statistical software. Results: The analyzed reference intervals of Osteocalcin by using Hoffmann method are from 8.8~39.4 ng/mL to 6.3~28.8 ng/mL for the case of the age from 20 to 30, from 7.7~31.9 ng/mL to 5.9~17.4 ng/mL for the case of the age from 31 to 40, and from 8.0~36.0 ng/mL to 5.5~20.1 ng/mL for the case of the age from 41 to 50, and from 8.0~50.5 ng/mL to 6.7~27.0 ng/mL for the case of the age from 51 to 60, and from 12.9~55.9 ng/mL to 7.5~27.5 ng/mL for the case of the age from 61 to 80. Reference intervals of Osteocalcin were not in agreement with those recommended by the manufacturers. Conclusions: Osteocalcin is used as an indication of metabolic bone diseases. So in our study we wanted to provide reference intervals of Osteocalcin that can be useful to a clinical decisions. Also, previous reference intervals should not be re-used and new intervals should be set by continuous analyzing.

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Temporal and Spatial Variation of Microalgal Biomass and Community Structure in Seawater and Surface Sediment of the Gomso Bay as Determined by Chemotaxonomic Analysis (색소분석을 통한 곰소만 내 해수와 퇴적물 중 미세조류 생체량과 군집구조의 시공간적 변화)

  • Lee, Yong-Woo;Park, Mi-Ok;Yoon, Ji-Hyun;Hur, Sung-Bum
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.2
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    • pp.87-94
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    • 2012
  • To compare monthly variations of phytoplankton biomass and community composition between in seawater and sediment of the Gomso Bay (tidal flat: approximately 75%), the photosynthetic pigments were analyzed by HPLC every month in 1999 and every two months in 2000. Ambient physical and chemical parameters (temperature, salinity, nutrients, dissolved oxygen, and chemical oxygen demand) were also examined to find the environmental factors controlling structure of phytoplankton community. The temporal and spatial variations of chlorophyll a concentration in seawater were correlated well with the magnitude of freshwater discharge from land. The biomass of microphytobenthos at the surface sediments was lower than that in other regions of the world and 2-3 times lower than phytoplankton biomass integrated in the seawater column. Based on the results of HPLC pigment analysis, fucoxanthin, a marker pigment of diatoms, was the most prominent pigment and highly correlated with chlorophyll a in seawater and sediment of the Gomso Bay. These results suggest that diatoms are the predominant phytoplankton in seawater and sediment of the Gomso Bay. However, the monthly variation of chlorophyll a concentration in seawater at the subtidal zone was not a good correlation with that in sediment of the Gomso Bay. Although pelagic plankton was identified in seawater by microscopic examination, benthic algal species were not found in the seawater. These results suggest that contribution from the suspended microphytobenthos in the tidal flat to the subtidal zone of the Gomso Bay may be low as a food source to the primary consumer in the upper water column of the subtidal zone. Further study needs to elucidate the vertical and horizontal transport magnitude of the suspended microphytobenthos in the tidal flat to the subtidal zone.

Effects of rrhGM-CSF on Morphology and Expression of PCNA in Regenerating Rat Liver (재생 중인 흰쥐 간의 형태학적 변화 및 PCNA 발현에 미치는 rrhGM-CSF의 영향)

  • Jeong, Jin-Ju;Heo, Si-Hyun;Kim, Ji-Hyun;Yoon, Kwang-Ho;Lee, Young-Jun;Han, Kyu-Boem;Kim, Wan-Jong
    • Applied Microscopy
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    • v.40 no.2
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    • pp.73-80
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    • 2010
  • Liver regeneration is a result of highly coordinated proliferation of hepatocytes and nonparenchymal liver cells. Partial hepatectomy (PH) is the most often used stimulus to study liver regeneration because, compared with other methods that use hepatic toxins, it is not associated with the tissue injury and inflammation, and the initiation of the regenerative stimulus is precisely defined. Granulocyte macrophage-colony stimulating factor (GM-CSF), which is a cytokine able to regulate the proliferation and differentiation of epithelial cells, was first identified as the most potent mitogen for bone marrow. Particularly, rrhGM-CSF, which is highly glycosylated and sustained longer than any other types of GM-CSF in the blood circulation, was specifically produced from rice cell culture. In this experiment, effects of rrhGM-CSF administration were evaluated in the regenerating liver after 78% PH of rats. Morphological changes induced by PH were characterized by destroyed hepatocyte plate around the central vein and enlarged nuclear cytoplasmic ratio and increased hepatocytes with two nuclei. And then, proliferation of liver cells (parenchymal and nonparenchymal) and rearrangement of plates and lobules seemed to be carried out during liver regeneration. These alterations in the experimental group preceded those of the control. Since proliferating cell nuclear antigen (PCNA) is known to be a nuclear protein maximally elevated in the S phase of proliferating cells, the protein was used as a marker of liver regeneration after PH in rats. PCNA levels by western blot analysis and immunohistology were compared between the two groups. PCNA protein expression of two groups at 12 hr and 24 hr after injury showed similar pattern. The protein expression showed the peak at 3 days in both groups, however, the protein level of the experimental group was higher than that of the control. On immunohistochemical observations, the reaction product of PCNA was localized at the nuclei of proliferating cells and the positive reaction in experimental group at 3 days was clearly stronger than that in control group. The results by Western blotting and immunohistology for PCNA showed similar pattern in terms of the protein levels. In conclusion, rrhGM-CSF administration during liver regeneration after 78% PH accelerated breakdown and restoration of the hepatic plate and expression of PCNA. These results suggest that rrhGM-CSF might play an important role during liver regeneration in rats.

Cytoprotective Effects of Schisandrin A against Hydrogen Peroxide-induced Oxidative Stress in SW1353 Human Chondrocytes (SW1353 인간 연골세포에서 산화적 스트레스에 대한 schisandrin A의 세포 보호 효과)

  • Jeong, Jin-Woo;Choi, Eun Ok;Kwon, Da Hye;Kim, Bum Hoi;Park, Dong Il;Hwang, Hye Jin;Kim, Byung Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1070-1077
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    • 2017
  • Chondrocyte apoptosis induced by reactive oxygen species (ROS) plays an important role in the pathogenesis of osteoarthritis. Schisandrin A, a bioactive compound found in fruits of the Schisandra genus, has been reported to possess multiple pharmacological and therapeutic properties. Although several studies have described the antioxidant effects of analogues of schisandrin A, the underlying molecular mechanisms of this bioactive compound remain largely unresolved. The present study investigated the cytoprotective effect of schisandrin A against oxidative stress (hydrogen peroxide [$H_2O_2$]) in SW1353 human chondrocyte cells. The results showed that schisandrin A preconditioning significantly inhibited $H_2O_2-induced$ growth inhibition and apoptotic cell death by blocking the degradation of poly (ADP-ribose) polymerase proteins and down-regulating pro-caspase-3. These antiapoptotic effects of schisandrin A were associated with attenuation of mitochondrial dysfunction and normalization of expression changes of proapoptotic Bax and antiapoptotic Bcl-2 in $H_2O_2-stimulated$ SW1353 chondrocytes. Furthermore, schisandrin A effectively abrogated $H_2O_2-induced$ intracellular ROS accumulation and phosphorylation of histone H2AX at serine 139, a widely used marker of DNA damage. Thus, the present study demonstrates that schisandrin A provides protection against $H_2O_2-induced$ apoptosis and DNA damage in SW1353 chondrocytes, possibly by prevention of ROS generation. Collectively, our data indicate that schisandrin A has therapeutic potential in the treatment of oxidative disorders caused by overproduction of ROS.

Diagnostic Usefulness of Serum Level of Cyfra 21-1, SCC Antigen and CEA in Lung Cancer (폐암에서 혈중 Cyfra 21-1, SCC 항원 및 CEA의 진단적 유용성)

  • Kim, Kyoung-Ah;Lee, Me-Hwa;Koh, Youn-Suck;Kim, Seon-Hee;Lim, Chae-Man;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Moon, Dae-Hyuk
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.6
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    • pp.846-854
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    • 1995
  • Background: Cytokeratin 19 is a subunit of cytokeratin intermediate filament expressed in simple epithelia such as respiratory epithelial cells and their malignant counterparts. An immunoradiometric assay is available to detect a fragment of the cytokeratin, referred to as Cyfra 21-1 in the serum. This study was conducted to evaluate the clinical utility of this new marker in the diagnosis of lung cancer compared with established markers of squamous cell carcinoma antigen (SCC Ag) and carcino-embryonic antigen(CEA). In addition, we compared the diagnostic sensitivity and specificity of Cyfra 21-1 with those of SCC Ag in squamous cell carcinoma of the lung. We also measured the level of Cyfra 21-1 in the different stages of squamous cell carcinoma of the lung. Method: We measured Cyfra 21-1(ELSA-CYFRA 21-1), SCC Ag(ABBOTT SCC RIABEAD) and CEA(ELSA2-CEA) in 79 patients with primary lung cancer and in 78 persons as a comparison group including 32 patients with pulmonary tuberculosis, 23 patients with benign lung disease and 23 cases with healthy individual. Cyfra 21-1 is measured by a solid-phase immunoradiometric assay(CIS Bio International, France) based on the two-site sandwich method. SCC Ag is measured by a radioimmunoassay(Abbott Laboratories, USA). CEA is measured by a immunoradiometric assay(CIS Bio International, France). All data were expressed as the mean$\pm$standard deviation. Results: 1) The mean value of Cyfra 21-1 was $18.38{\pm}3.65\;ng/mL$ in the lung cancer and $1.l6{\pm}0.53\;ng/mL$ in the comparison group(p<0.0001). SCC Ag was $3.53{\pm}6.06\;ng/mL$ in the lung cancer and $1.19{\pm}0.5\;ng/mL$ in the comparison group(p<0.01). CEA was $35.03{\pm}13.9\;ng/mL$ in the lung cancer and $2.89{\pm}1.01\;ng/mL$ in the comparison group(p<0.0001). 2) Cyfra 21-1 level in squamous cell carcinoma($31.52{\pm}40.13\;ng/mL$) was higher than that in adenocarcinoma($2.41{\pm}1.34\;ng/mL$)(p<0.0001) and small cell carcinoma($2.15{\pm}2.05\;ng/mL$)(p=0.007). SCC Ag level in squamous cell carcinoma($5.1{\pm}7.68\;ng/mL$) was higher than that in adenocarcinoma($1.36{\pm}0.69\;ng/mL$)(p=0.009) and small cell carcinoma($1.1{\pm}0.24\;ng/mL$) (p=0.024). 3) The level of Cyfra 21-1 was not correlated with the progression of stage in squamous cell carcinoma of the lung. 4) Using the cut-off value of 3.3ng/mL, the diagnostic sensitivity of Cyfra 21-1 was 83% in squamous cell carcinoma, 22% in adenocarcinoma and 17% in small cell carcinoma. The sensitivity of SCC Ag and CEA were 39% and 20%, respectively in squamous cell carcinoma, 11% and 39% in adenocarcinoma, and 0% and 33% in small cell carcinoma. 5) Comparison of the receiver operating characteristics curves(ROC curve) for Cyfra 21-1, SCC Ag and CEA revealed that Cyfra 21-1 showed highest diagnostic sensitivity among them in the diagnosis of lung cancer. Conclusion: Cyfra 21-1 is thought to be a better tumor marker for the diagnosis of lung cancer than SCC Ag and CEA, especially in squamous cell carcinoma of the lung.

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Effect of combined exercise on metabolic bio-marker in overweight and obese children (복합운동이 과체중 및 비만 남자 초등학생의 대사적지표에 미치는 영향)

  • Kim, Hyun-Jun;Kim, Tae-Un;Lee, Sangyeoup;Shin, Goon-Soo;Kim, Young-Joo;Kim, Su-Yung
    • Clinical and Experimental Pediatrics
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    • v.49 no.9
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    • pp.946-951
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    • 2006
  • Purpose : The purpose of this study was to demonstrate the effectiveness of combined exercise for 12 weeks on the adiponectin and obesity related variables in overweight and obese children. Methods : Eighteen children in 5th grade in a certain elementary school in Busan were recruited. They were all overweight or obese children(more than 85 percentile in body mass index). Nine children in the experimental group were given exercises consisting of walking and band resistant training for 12 weeks. Auxological data(including height, weight and body fat mass) and laboratory data (fasting blood sugar, insulin, adiponectin) were checked at baseline and at the 1 week, and at the 4 weeks and 12 weeks stages of their exercise program. Insulin resistance and sensitivity were evaluated indirectly using HOMA index and QUICKI index. Results : Adiponectin gradually decreased until the 4 weeks point and gradually increased thereafter to the starting level at the 12 weeks stage. Body weight, body mass index(BMI) and HOMA index significantly decreased more at the 1 week, 4 weeks, and 12 weeks stages in the experimental group than in the control group. Body fat mass significantly decreased at 12 weeks. The change of insulin was significantly correlated with changes of body weight and BMI. But there was no correlation between changes of adiponectin and changes of insulin. Conclusion : Exercise seems to effect the adiponectin concentration. And it might be assumed that exercise increases the adiponectin concentration if it is continued for long time(may be more than 12 weeks). More studies may be necessary to draw that conclusion.

Analysis of ethyl glucuronide (EtG) in Hair for the diagnosis of chronic alcohol abuse of Korean (한국인의 만성 알코올 중독 진단을 위한 모발에서 Ethyl Glucuronide (EtG) 분석법 연구)

  • Gong, Bokyoung;Jo, Young-Hoon;Ju, Soyeong;Min, Ji-Sook;Kwon, Mia
    • Analytical Science and Technology
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    • v.33 no.3
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    • pp.151-158
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    • 2020
  • Alcohol, which can easily be obtained in the same way as ordinary beverages, is harmful enough to cause death due to excessive drinking and chronic alcohol intake, so it is important to maintain a proper amount of drinking and healthy drinking habits. In addition, the incidence of behavioral disturbances and impaired judgments that can be caused by chronic alcohol drinking of more than adequate amounts of alcohol is also significant. Accordingly it is very useful for forensic science to check whether the person involved is drunken or is alcoholism state in various accidents. Currently, in Korea, alcohol consumption is determined by detecting the level of alcohol or alcohol metabolism 'ethyl glucuronide (EtG)' in blood or urine samples. However, analysis of alcohol or EtG in blood or urine can only provide information about the current state of alcohol consumption because of a narrow window of detection time. Therefore, it is important to analyze the EtG as a long-term direct alcohol metabolite bio-marker in human hair and to investigate relationship between alcohol consumption and EtG concentration for the evaluation of chronic ethanol consumption. In this study, we established an analytical method for the detection of EtG in Korean hair efficiently and validated selectivity, linearity, limits of detection (LOD), limits of quantification (LOQ), matrix effect, recovery, process efficiency, accuracy and precision using liquid chromatography tandem mass spectrometry (LC-MS/MS). In addition, the assay performance was evaluated in Korean social drinker's hair and the postmortem hair of a chronic alcoholism. The results of this study can be useful in monitoring the alcohol abuse of Korean in clinical cases and legal procedures related to custody and provide a useful tool to evaluate postmortem diagnosis of alcoholic ketoacidosis in forensics.

Effect of Keratinocyte Derived Exosome on Proliferation and Migration on Human Skin Keratinocyte (각질형성세포 유래 엑소좀이 피부각질형성세포의 증식과 이주에 미치는 영향)

  • Kim, Do Yoon;Yu, Ho Jin;Hwang, Dae Il;Jang, Sang Hee;Lee, Hwan Myung
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.4
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    • pp.359-366
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    • 2016
  • Exosome, a small vesicle secreted from cells, has diverse functions depending on cell origins and tissue types and plays a important role in cell viability and intercellular communication. Recently, many researchers have demonstrated the use of exosomes for the treatment of cancers and immune diseases, and the development of diagnostic biomarker. However, the secretion mechanism of exosome from skin cell and its physiological functions in skin remain unclear. Thus, this study aimed to explore whether keratinocyte-derived exosome affects proliferation and migration in HaCaTs. Exosomes were isolated from HaCaTs by ExoQuick-TC and then boiled or unbolied. Boiled and unboiled exosome induced proliferation in HaCaTs in a dose-dependant manner ($0.1{\sim}20{\mu}g/mL$), respectively. Boiled and unboiled exosome at concentration of $20{\mu}g/mL$ increased proliferation level in HaCaTs by $186.96{\pm}3.87%$ and $193.48{\pm}10.48%$ compared with control group. Unboiled exosome stimulated migration in HaCaTs in a dose-dependent manner ($0.1{\sim}20{\mu}g/mL$), which reached a maxium at concentration of $20{\mu}g/mL$ ($179.39{\pm}4.89%$ of control), but boiled exosome did not affect HaCaT migration. In addition, unboiled exosome ($0.1{\sim}20{\mu}g/mL$) dose-dependently stimulated sprout outgrowth in HaCats. These results demonstrate that in exosome from HaCaTs, heat-stable components such as lipid may induce HaCaT proliferation and heat-unstable components such as protein may stimulate migration and sprout outgrowth in HaCaTs, thereby leading to reepithelialization and skin-wound healing activities. It is concluded that exosomes from HaCaTs may be used as cosmetic materials.