• Title/Summary/Keyword: Transcription Culture

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EXPRESSION AND FUNCTIONAL CHARACTERIZATION OF ODONTOBLAST-DERIVED GENE: OD314 (상아모세포 관련 유전자, OD314의 발현과 기능 연구)

  • Kim, Doo-Hyun;Kim, Heung-Joong;Jeong, Moon-Jin;Son, Ho-Hyun;Park, Joo-Cheol
    • Restorative Dentistry and Endodontics
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    • v.29 no.4
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    • pp.399-408
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    • 2004
  • Odontoblasts are responsible for the formation and maintenance of dentin. They are known to synthesize unique gene products including dentin sialophosphoprotein (DSPP). Another unique genes of the cells remain unclear. OD314 was isolated from the odontoblasts/pulp cells of rats and partially characterized as an odontoblast-enriched gene (Dey et al., 2001). This study aimed to elucidate the biological function of OD314, relating to odontoblast differentiation and dentinogenesis. After determining the open reading frame (ORP) of OD314 by transient transfection analysis using green fluorescent protein (GPP) expression vector, mRNA in-situ hybridization, immunohistochemistry, reverse transcription-polymerase chain reaction (RT-PCR) and western analysis were performed. The results were as follows: 1. In in-situ hybridization, OD314 mRNAs were expressed in odontoblasts of developing coronal and root pulp. 2. OD314 was a novel protein encoding 154 amino acids, and the protein was mainly expressed in cytoplasm by transient transfection analysis. 3. Mineralized nodules were associated with multilayer cell nodules in the culture of human dental pulp cells and first detected from day 21 using alizarin-red S staining. 4. In RT-PCR analysis, OD314, osteocalcin (OC) and DSPP strongly expressed throughout 28 days of culture. Whereas, osteonectin (ON) mRNA expression stayed low up to day 14, and then gradually decreased from day 21. 5. Western blots showed an approximately 17 kDa band. OD314 protein was expressed from the start of culture and then increased greatly from day 21. In conclusion, OD314 is considered as an odontoblast-enriched gene and may play important roles in odontoblast differentiation and dentin mineralization.

A Study on Okso, Keun Seob's Hakchangeui(학창의)-Examining Data about Hakchangeui Written in $\lceil$Oksoko(옥소고)$\rfloor$- (옥소 권섭의 학창의에 관한 연구 - "옥소고"소재 학창의 관계자료를 대상으로-)

  • Lee Min-Joo
    • The Research Journal of the Costume Culture
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    • v.13 no.2 s.55
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    • pp.269-279
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    • 2005
  • Okso, Keun Seob($1671\sim1759$) was descended from a noble family of Noreun(노론) and learned from his uncle, Keun Sang-ha who is a disciple of Song Si-Yeol, Kim Chang-hyeub and Kim Chang-heub. His $\lceil$Oksoko$\rfloor$ remains in the form of a transcription or a lithographic printing, and documents on 'Hakchangeui' are recorded in jabeui(잡의), Euijedogi(의제도기), which is a part of it. He left Seoul in 1714, settled in Chungpung, journeyed to many places, including the Hwang river area and Jecheon and led a literary and artistic life as Sadaebu(사대부), which was written in a collection of his works where the record about 'Hakchangeui', the most suitable clothes for the life of a retired scholar remains today to express one part of his life. This study introduces data about Hakchangeui written in $\lceil$Oksoko$\rfloor$, pursues the reason why Okso, Keun Seob told about it and intends to embody the figure of Hakchangeui based on the system, measure, material, and sewing method according to the records in a collection. Hakchaneui described in $\lceil$Oksoko$\rfloor$ reveals a structure that is different from old Hakchaneui, showing characteristics like the following. It is a Sadaebu's costume for field amusement in which the length of a former part is longer than that of a later part and the side seam of a garment rips. Black Yeon is attached to the white texture. Yeon widths of Git(collar), Sugu and Gil are varied, respectively. Yeon's figure attached to Gil and Somaejindong is a mountain. Git is Bangryeong and link parts between Git and Gil is connected with Sangchim. This Hakchangeui is considered to playas a gown because it has no string or support to adjust his dress. Especially we can identify that Hwayanggun(화양건) was used in this Hakchangeui.

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In vitro characterization of human dental pulp stem cells isolated by three different methods

  • Jang, Ji-Hyun;Lee, Hyeon-Woo;Cho, Kyu Min;Shin, Hee-Woong;Kang, Mo Kwan;Park, Sang Hyuk;Kim, Euiseong
    • Restorative Dentistry and Endodontics
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    • v.41 no.4
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    • pp.283-295
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    • 2016
  • Objectives: In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. Materials and Methods: HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting. Results: Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups. Conclusions: Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

Effect of cell-penetrating peptide-conjugated estrogen-related receptor ${\beta}$ on the development of mouse embryos cultured in vitro

  • Yang, Ning Jie;Seol, Dong-Won;Jo, Junghyun;Jang, Hyun Mee;Yoon, Sook-Young;Lee, Dong Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.1
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    • pp.1-8
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    • 2014
  • Objective: Estrogen related receptor ${\beta}$ (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotencyrelated genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development. Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without $2{\mu}g/mL$ CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry. Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group. Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos' expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.

Post-pandemic influenza A (H1N1) virus detection by real-time PCR and virus isolation

  • Zaki, Ali Mohamed;Taha, Shereen El-Sayed;Shady, Nancy Mohamed Abu;Abdel-Rehim, Asmaa Saber;Mohammed, Hedya Said
    • Korean Journal of Microbiology
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    • v.55 no.1
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    • pp.25-32
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    • 2019
  • Influenza A (H1N1) virus caused a worldwide pandemic in 2009-2010 and still remains in seasonal circulation. Continuous surveillance activities are encouraged in the post pandemic phase to watch over the trend of occurrence every year, this is better to be done by a rapid and sensitive method for its detection. This study was conducted to detect proportions of occurrence of influenza A virus (H1N1) in patients with influenza-like illness. Samples from 500 patients with influenza or influenza-like clinical presentation were tested by real-time reverse transcription polymerase chain reaction (RT-PCR) and virus tissue culture. Among the total 500 participants, 193 (38.6%) were females and 307 (61.4%) males. Seventy-one patients (14.2%) were positive for H1N1 virus infection with real-time RT-PCR while 52 (10.4%) were positive by tissue culture. Non-statistically significant relation was found between age and gender with the positivity of H1N1. Sensitivity and specificity of real-time RT-PCR was 98.08% and 95.54%, respectively, in comparison to virus isolation with accuracy 95.8%. This study showed that H1N1 virus was responsible for a good proportion of influenza during the post-pandemic period. Real-time RT-PCR provides rapidity and sensitivity for the detection of influenza A virus (H1N1) compared with virus isolation and thus it is recommended as a diagnostic tool.

A Study on the Gold Foil Patch Design Using Traditional Patterns (전통문양을 활용한 금박패치디자인 연구)

  • Oh, Yu-Kyeong;Song, Jung-A
    • Fashion & Textile Research Journal
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    • v.24 no.1
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    • pp.95-107
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    • 2022
  • In reality, the distinction between the Korean traditional culture and the cultures of other countries is at a delicate boundary. Additionally in the wake of the recent socio-cultural confrontation between Korea and China, it has become necessary to establish the foundation and area of Korean traditional culture and to actively utilize the importance of improving awareness of Korean traditional culture. To reorganize the reckless use of the gold foil pattern shown in the rental hanbok, data on the museum's collection of gold leaf patterns were collected and analyzed. Based on the gold foil, Gilsang characters such as Phoenix pattern, Crane pattern, Bat pattern, Flower pattern, Fruit pattern and recovery advice were extracted through references. The traditional gold foil pattern was reconstructed and relocated to design the gold leaf patch. Based on the collection and analysis of the museum's relics, the Wonsam & Daedae, Dangeui, Sranchima, Sagyusam, Jeonbok, Bokgun, and Daenggi were produced. Therefore, we present the possibility of producing gold foil and modern methods for producing gold foil using laser cutting techniques that can express refinement and complexity well, and gold foil thermal transfer paper with retouchable effects. Additionally, we would like to reflect upon the practicality and the convenience to modern people by considering the complexity and hassle of the traditional gold foil production process, and the disadvantages of processes that require relatively longer time. It intends to help revitalize the market of Korean traditional clothing and fashion products.

Growth factors improve the proliferation of Jeju black pig muscle cells by regulating myogenic differentiation 1 and growth-related genes

  • Park, Jinryong;Lee, Jeongeun;Song, Ki-Duk;Kim, Sung-Jo;Kim, Dae Cheol;Lee, Sang Cheol;Son, Young June;Choi, Hyun Woo;Shim, Kwanseob
    • Animal Bioscience
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    • v.34 no.8
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    • pp.1392-1402
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    • 2021
  • Objective: The growth rate of pigs is related to differentiation and proliferation of muscle cells, which are regulated by growth factors and expression of growth-related genes. Thus, the objective of this study was to establish optimal culture conditions for Jeju black pig (JBP) muscle cells and determine the relationship of various factors involved in muscle growth with the proliferation of JBP muscle cells. Methods: Muscles were taken from the femur skeletal muscle of JBP embryos. After isolation of the muscle cells, cells were cultured in a 6-well plate under four different culture conditions to optimize culture conditions for JBP muscle cells. To analyze proliferation rate of JBP muscle cells, these muscle cells were seeded into 6-well plates at a density of 1.5×105 cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (MyoD) expression and growth-related gene expression in JBP muscle cells, respectively. Results: We established a muscle cell line from JBP embryos and optimized its culture conditions. These muscle cells were positive for MyoD, but not for paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (EGF+bFGF) than that without a growth factor or containing EGF alone. Treatment with EGF and bFGF significantly induced the expression of MyoD protein, an important transcription factor in muscle cells. Moreover, we checked the changes of expression of growth-related genes in JBP muscle cells by presence or absence of growth factors. Expression level of collagen type XXI alpha 1 gene was changed only when EGF and bFGF were added together to culture media for JBP muscle cells. Conclusion: Concurrent use of EGF and bFGF increased the expression of MyoD protein, thus regulating the proliferation of JBP muscle cells and the expression of growth-related genes.

The Effects of Dictamni Radicis Cortex Extracts on DNCB Induced Atopic Dermatitis-like Skin Lesions of NC/Nga Mouse (백선피(白鮮皮) 추출물이 DNCB로 유발된 NC/Nga 생쥐의 아토피 피부염에 미치는 영향)

  • Lee, Youn Shil;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.28 no.4
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    • pp.1-29
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    • 2014
  • Objectives Dictamni Radicis Cortex extracts (DRC) has been known to suppress allergic reaction, however the cellular target of DRC and its mode of action remain unclear. The purpose of this study is to investigate the effects of Dictamni Radicis Cortex extracts on DNCB induced atopic dermatitis-like skin lesions of NC/Nga mouse. Methods This study was designed to investigate the effects of DRC extract in the DNP-IgE-induced activation of MC/9 murine mast cell lines in vitro and in the DNCB-induced activation of NC/Nga mouse in vivo. For this investigation, We examined IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ and GM-CSF mRNA expression by Real-time PCR, IL-13, MIP-$1{\alpha}$ production by ELISA analysis and manifestations of NFAT1, NFAT2, AP-1 and NF-${\kappa}B$ p65 transcription factors by western blotting in vitro. Then, we examined WBC, eosinophil and neutrophil in NC/Nga mouse, IL-5, IL-13 in serum, IFN-${\gamma}$, IL-4 in the spleenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $^+Gr-1^+CD11b$, $B220^+CD23^+$ in the ALN, PBMCs and dorsal skin, IL-5, IL-13 in the dorsal skin by Real-time PCR and the distribution of mast cells by H&E and toluidine blue. Results In vitro the mRNA expression of IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$, GM-CSF and IL-13, MIP-$1{\alpha}$ production by ELISA analysis were completely abolished by DRC and the western blot analysis decreased the expression of mast cell-specific transcription factors including NFAT-1, NF-${\kappa}B$ p65. In vivo DRC oral adminstration also decreased the counts of WBC, eosinophils and inflammatory cytokines such as IL-13 and IgE in the serum. DRC oral adminstration elevated IL-4 level in the spleenocyte culture supernatant. DRC oral adminstration decreased total ALN cells, total skin cells, cell numbers of $CD4^+$, $B220^+CD23^+$ in the ALN, $^+Gr-1^+CD11b$ in the PBMCs and $CD4^+$, $CD8^+$ in the dorsal skin. The mRNA expression of IL-5, IL-13, thickness of epidermis, inflammation immune cells and mast cells were abolished by DRC in the dorsal skin. Conclusions Histological examination showed that infiltration levels of immune cells in the skin of AD-induced NC/Nga mouse were much improved by DRC oral adminstration. These results, therefore, suggest that DRC can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis induced in NC/Nga mouse, and may play an important role in recovering AD symptoms.

The Effects of Prunus Armeniaca Linne Var Fractions on Th2 Cytokine Expression and Atopic Dermatitis of NC/Nga Mouse (행인(杏仁) 분획물이 Th2 cytokine 발현과 NC/Nga mouse의 아토피 피부염에 미치는 영향)

  • Kang, Ki Yeon;Han, Jae Kyung;Kim, Yun Hee
    • The Journal of Pediatrics of Korean Medicine
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    • v.30 no.4
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    • pp.29-59
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    • 2016
  • Objectives PRAL (Prunus armeniaca Linne Var) has been known to suppress allergic reaction. However, the cellular target and its mechanism of action were unclear. This study was designed to investigate the effect of PRAL on RBL-2H3 mast cell, which is PMA-Ionomycin-induced activated in vitro and the effect of PRAL on the MNC/Nga mice that are DNCB-induced activated in vivo. Methods In this study, IL-4, IL-13 production were examined by ELISA analysis; IL-4, IL-13, IL-31, IL-31Ra, $TNF-{\alpha}$ and GM-CSF mRNA expression were examined by Real-time PCR; manifestations of AP-1 and MAPKs transcription factors were examined by western blotting in vitro. Then skin rashes have been evaluated and verified the distribution of mast cells by H&E and toluidine blue. Also, WBC, eosinophil and neutrophil, IgE level in serum, $IFN-{\gamma}$, IL-4, IL-5 in the splenocyte culture supernatant, the absolute cell numbers of $CD4^+$, $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$, $CD3^+CD69^+$ in the Axillary Lymph Node (ALN), PBMCs and dorsal skin and IL-5, IL-13, IL-31, IL-31Ra in the dorsal skin by Real-time PCR were all evaluated from the NC/BNga mice. Results As a result of this study, the mRNA expression of IL-4, IL-13, IL-31, IL-31Ra and $TNF-{\alpha}$ and IL-4, IL-13 production, shown in ELISA analysis, were suppressed by PRAL. Results from the western blot analysis showed decrease on the expression of mast-cell-specific transcription factors, including AP-1 and p-JNK, p-ERK. Histological examination showed that infiltration levels of immune cells in the skin of the AD-induced NC/Nga mice were improved by PRAL orally adminstration. Orally- administered PRAL group also showred decreased level of IgE in the serum. This group has shown decreased the level of IL-4, IL-5, but shown elevated $IFN-{\gamma}$ level in the splenocyte culture supernatant. The same group also has shown decreased cell numbers of $CD4^+$, $CD8^+$, $CD3^+CD69^+$ in the ALN, and $CD4^+$, $Gr-1^+CD11b^+$ in the dorsal skin. PRAL oral adminstration increased cell numbers of $CD4^+$, but decreased cell numbers of $CD8^+$, $Gr-1^+CD11b^+$, $B220^+CD23^+$ in the PBMCs. Conclusions Obtained results suggest that PRAL can regulate molecular mediators and immune cells that are functionally associated with atopic dermatitis (AD) induced in the NC/Nga mice. This may play an important role in recovering AD symptoms and suppressing pruritus.

The Study on Vitrification and Ultrarapid Thawing of Human Embryonic Stem Cells (인간 배아 줄기세포의 초자화 동결 및 초급속 융해에 관한 연구)

  • Moon, Shin-Yong;Park, Yong-Bin;Kim, Hee-Sun;Sung, Ki-Chung;Oh, Sun-Kyung;Chun, Dae-Woo;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.13-20
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    • 2002
  • Objective: This study was carried out to establish the effectiveness of the vitrification method and the optimal cryoprotectants in the cryopreservation of human embryonic stem cells (ESC). Materials and Methods: Human ESC clumps established at Seoul National University Hospital (SNUhES 1) were cryopreserved with the vitrification method using the EM grid. EDS and EFS40 were used as vitrification solutions. Results: Between the EDS and EFS40 groups, there was no significant difference in the recovery rate after cryopreservation of human ESC. The formation rates of ESC colonies in the vitrified groups were significantly lower than those in the control ESC group (p<0.05, p<0.05). In addition, the formation rate of ESC colonies in the EDS group was significantly higher than that in the EFS40 group (p<0.05). The ESC colonies in the vitrified groups were significantly smaller after culture duration of 2 and 4 days, respectively, compared with the control ESC group (p<0.1, p<0.05). However, these effects could be reduced to nonsignificant level by the additional culture of ESC colonies. The vitrified human ESC retained the properties of pluripotent cells, including the expression of cell surface. markers for the undifferentiated cells such as alkaline phosphatase and SSEA-4 (stage-specific embryonic antigen-4), and the expression of transcription factor Oct-4 (octamer-binding transcription factor-4), and the normal karyotype. Conclusion: The vitrification method using the EM grid and EDS solution was confirmed to be very effective for the cryopreservation of human ESC.