• Title/Summary/Keyword: Polymerase Chain Reaction(PCR)

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Isolation and characterization of human dental tissue-derived stem cells in the impacted wisdom teeth: comparison of dental follicle, dental pulp, and root apical papilla-derived cells (미성숙 매복지치의 치낭, 치수, 치근유두 조직에서 다능성 줄기세포의 분리와 특성화에 대한 연구)

  • Song, Jung-Ho;Park, Bong-Wook;Byun, June-Ho;Kang, Eun-Ju;Rho, Gyu-Jin;Shin, Sang-Hun;Kim, Uk-Kyu;Kim, Jong-Ryoul
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.36 no.3
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    • pp.186-196
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    • 2010
  • Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

Association between the Human Surfactant Protein-A(SP-A) Gene Locus and Chronic Obstructive Pulmonary Disease in Korean Population (한국인에서 만성폐쇄성폐질환과 인체 폐 표면 활성제 단백-A 유전자 대립형질의 상관관계)

  • Na, Joo Ock;Oh, Myung Ho;Choi, Jae Sung;Seo, Ki Hyun;Kim, Yong Hoon
    • Tuberculosis and Respiratory Diseases
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    • v.60 no.6
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    • pp.638-644
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    • 2006
  • Backgrounds: This study investigated whether or not a polymorphism in the gene encoding the surfactant protein A(SP-A) has any bearing on the individual susceptibility to the development of chronic obstructive pulmonary disease(COPD) in a genetically homogenous Korean population. Methods: The genotypes of 19 COPD patients and 20 healthy neonates as controls were tested using a polymerase chain reaction followed by restriction fragment length polymorphism analysis for the SP-A gene. Results: The specific frequencies of the 6A2 and 6A18 alleles of SP-A1 and the 1A2 allele of SP-A2 were much higher in the COPD group than control group (p<0.05). However, the frequencies of the 6A3 and 6A4 alleles of SP-A1 and the 1A0 allele of SP-A2 in the COPD group were significantly lower than the control group. In the COPD group, the frequencies of the +50 locus genotypes GG of SP-A1 and the +9 locus genotypes CC of SP-A2 were 85.0% and 60.6%, respectively, and 19.7% and 24.8% in the control group, respectively. The frequencies of the polymorphic genotypes or alleles showed a statistically significant difference between the COPD group and the control group (P<0.05). Conclusion: A genetic polymorphism in SP-A is associated with the development of COPD in the Korean population.

Comparison of Stress Response in Diallel Crossed Korean Domestic Chicken Breeds (토종 종계를 이용한 이면 교배조합 계통 간 스트레스 반응정도 비교 분석)

  • Cho, Eun Jung;Park, Ji Ae;Choi, Eun Sik;Sohn, Sea Hwan
    • Korean Journal of Poultry Science
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    • v.43 no.2
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    • pp.77-88
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    • 2016
  • To establish a new synthetic Korean meat chicken breed, we tested $5{\times}5$ diallel cross mating experiment with domestic chicken breeds. Comparing stress responses among diallel crossed chicken breeds, we analyzed telomere length, DNA damage and expressions of heat shock protein genes (HSPs) as the markers of the stress response. The telomere length was measured by quantitative fluorescence in situ hybridization on the nuclei of lymphocytes. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes were analyzed by quantitative real-time polymerase chain reaction in lymphocytes. The DNA damage rate of lymphocytes was quantified by the comet assay known as the single cell gel electrophoresis. In results, there were significant differences in the values of the stress markers such as telomere length, HSPs and DNA damage rate, and also were significant differences in viabilities and body weights among the $5{\times}5$ diallel crossed chicken breeds. The telomere shortening rate, expression values of HSPs and DNA damage rate were significant low in W and Y crossed chickens compare to the others, but GG pure breed showed the highest values in the 25 crossed chickens. Estimating correlation coefficient, the survival rate positively correlated to telomere length, but negatively correlated to the expression levels of HSP-70, $HSP-90{\alpha}$, $HSP-90{\beta}$ genes and to the value of % DNA in tail as DNA damage rate. The expression levels of HSP-70, $HSP-90{\alpha}$ and $HSP-90{\beta}$ genes of dead chickens had significantly higher than those of survival chickens. According to the results on the stress marker analysis, it would be considered that the crossed breeds had more stress resistant than the pure breeds, and the crossed chickens with a light strain such as W or Y were relatively resistant to stress, but the crossed chickens with a heavy strain such as G, H, F were susceptible to stress.

Analysis of Palivizumab Prophylaxis in Patients with Acute Lower Respiratory Tract Infection Caused by Respiratory Syncytial Virus (Respiratory syncytial virus로 인한 급성 하기도 감염 입원 환자에서 Palivizumab 예방요법 유무에 따른 비교 분석)

  • Min, Sung Ju;Song, Jung Sook;Choi, Jang Hwan;Seon, Han Su;Kang, Eun Kyeong;Kim, Do Hyun;Kim, Hee Sup
    • Pediatric Infection and Vaccine
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    • v.18 no.2
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    • pp.154-162
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    • 2011
  • Purpose : The aim of this study was to identify the clinical characteristics of lower respiratory tract infection due to respiratory syncytial virus (RSV) in young children and to provide information for an effective guideline for palivizumab administration in Korea. Methods : We reviewed medical charts of 167 patients under 3 years of age who were hospitalized in Dongguk University Ilsan Hospital for lower respiratory tract infection between January 2007 and February 2011. Diagnosis of the virus was made based on the multiplex real time polymerase chain reaction. Results : There were 113 patients who were infected by respiratory syncytial virus. 90 patients were term infants and 23 patients were preterm infants. No difference was shown between term and preterm infants except the days of admission which was 9.0${\pm}$6.0 days and 12.6${\pm}$21.0 days respectively. In the preterm group their mean age at the time of admission was 5.21${\pm}$4.9 months and the mean gestational age was 33.1${\pm}$4.3 weeks, and the mean birth weight was 2,152${\pm}$950 g. Only 4 patients were born under 28 weeks gestational age and were candidates for palivizumab administration. Conclusion : Most of the patients with severe RSV lower respiratory tract infection were term or near term infants who were not candidates for palivizumab prophylaxis. A nationwide study is needed to make a new risk stratified guideline for RSV prophylaxis for our country.

Whitening Activity of Sambucus Sieboldiana Var. Pendula (Nakai) Extract (말오줌나무 추출물의 미백활성 검증)

  • Yoo, Dan-Hee;Kim, Jin-Tae;Oh, Min-Jeong;Yeom, Hyeon-Ji;Lee, Jin-Young
    • Journal of Life Science
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    • v.29 no.3
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    • pp.279-286
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    • 2019
  • This study evaluated the anti-oxidant and whitening effects of a 70% ethanol extract of the Sambucus sieboldiana var. pendula (Nakai) (SS). At $1,000{\mu}g/ml$ concentration, the electron donating ability of this SS extract was found to be 86.21% and the ABTS+ radical scavenging ability was 97.9%. In terms of whitening activity, the tyrosinase inhibitory effect of the extract was 37%, also at $1,000{\mu}g/ml$ concentration. To explore the extractefftoxicity to B16F10 melanoma cells, a 3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl-tetrazoliumbromide assay was performed. Results showed 90% or more cells remained viable at $100{\mu}g/ml$ concentration. A Western blot of the SS extract was used to measure microphthalmia-associated transcription factor (MITF), tyrosinase-related protein-1 (TRP-1), tyrosinase relate protein-2 (TRP-2), and the tyrosinase protein expression inhibitory effect at 25, 50, $100{\mu}g/ml$ concentrations; ${\beta}-actin$ was used as a positive control. Consequently, the MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect were seen to decrease by 34.5%, 45.6%, 58.4%, and 79.6%, respectively, at $100{\mu}g/ml$ concentration. These were also then measured by reverse transcription-polymerase chain reaction at 25, 50, $100{\mu}g/ml$ concentrations with GAPDH as a positive control. As a result, the SS extract was seen to decrease MITF, TRP-1, TRP-2, and the tyrosinase protein expression inhibitory effect by 85.4%, 67.5%, 85.2%, 67.1%, respectively at the $100{\mu}g/ml$ concentration. We therefore confirmed the possibility of Sambucus sieboldiana var. pendula (Nakai) extract as a whitening material.

Comparative Analysis on Antioxidant Activity in Various Human Skin Fibroblasts and Mesenchymal Stem Cells (사람의 피부 섬유아세포 및 중간엽 줄기세포에서 항산화 활성의 반응에 대한 비교 분석)

  • Kong, Ji-Weon;Park, Ryeok;Park, Joon-Woo;Lee, Joo-Yeong;Choi, Yeon-Joo;Moon, Sun-Ha;Kim, Hyeon-Ji;Jeon, Byeong-Gyun
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.2
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    • pp.394-406
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    • 2019
  • The cellular senescence may be due to damage by the reactive oxygen species (ROS). This study has compared the antioxidant activity in the human cell lines of various origins, including 10S and 50S-derived normal skin fibroblasts, and 10S bone marrow, dental tissue and adipose-derived adult stem cells. After being exposed to $H_2O_2$, half inhibitory concentration ($IC_{50}$) values by cytotoxicity assay was significantly (P<0.05) lower in 50S-derived skin fibroblasts, than in 10S-derived skin fibroblasts and various adult stem cell lines. The cell population doubling time (PDT) and the cell frequency with high senescence associated-${\beta}$-galactose activity were remarkably increased in 50S-derived fibroblasts exposed to 50 ppm $H_2O_2$ for 7 days, than those of 10S-derived fibroblasts and various adult stem cell lines. Further, the expression level of antioxidant-related genes, glutathione peroxidase (GPX) and catalase (CAT), was investigated in 10S and 50S-derived skin fibroblasts, and 10S-derived various adult stem cells by reverse transcription polymerase chain reaction (RT-PCR). The expression level of GPX was higher in most of cell lines, compared to CAT, and a significantly (P<0.05) higher expression level of GPX was observed in 10S-derived skin fibroblasts and adult stem cell lines, compared to 50S-derived skin fibroblasts. We concluded that old-aged skin fibroblasts seemed to be less resistant against ROS than young-aged skin fibroblasts and adult stem cells.

Moderate Intensity Exercise Has More Positive Effects on The Gene Expression of Inflammasome, M1, M2 Macrophage Infiltration and Brown Adipocyte Markers Compared to High Intensity Exercise in Subcutaneous Adipose of Obese Mice Induced By High Fat Diet (비만모델에서 중강도 운동에 의한 인플라마좀, 대식세포 침윤, 갈색지방 관련 바이오 마커의 개선 효과)

  • Kim, Yong-An;Pitriani, Pipit;Park, Hee-Geun;Lee, Wang-Lok
    • Journal of Life Science
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    • v.29 no.3
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    • pp.303-310
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    • 2019
  • The purpose of the study was to compare the effect of either moderate or high intensity aerobic exercise on inflammasome, M1, M2 macrophage infiltration and brown adipocyte markers in subcutaneous adipose tissue of the high fat diet-induced obese mice. The 4 weeks male C57BL/6 mice were randomly assigned to four groups: normal diet control (NC; n=10), high-fat diet control (HC; n=10), high fat diet with moderate intensity exercise (HME; n=10), or high fat diet with high intensity exercise (HIE; n=10) groups. The high fat diet was given 60% calories from fat whereas normal diet was given 18% calories from fat. The moderate intensity exercise group (HME) was set at 10m/min in the first 2 weeks, 12m/min in 3-5 weeks and 14m/min in 6-16 weeks and the high intensity exercise group (HIE) was set at 14m/min in the first 2 weeks, 17m/min in 3-5 weeks and 18m/min in 6-16 weeks. The semi quantitative reverse transcription-polymerase chain reaction (RT PCR) was used to analyze the gene expression. The moderate intensity exercise significantly reduced the expression of NLRP3, F480, CD11c and CD86. Further, the moderate intensity exercise significantly increased CD206 and $PGC1{\alpha}$, BMP7 and PRDM. The high intensity exercise significantly reduced NLRP3, CD11c and CD86. Further, the high intensity exercise significantly increased $PGC1{\alpha}$ and BMP7. In conclusion, moderate intensity exercise has more positive effects on inflammasome, M1, M2 macrophage infiltration and brown adipocyte maskers compared to high intensity exercise in high fat diet induced obese mice.

Association Study of Zygote Arrest 1 on Semen Kinematic Characteristics in Duroc Boars (두록 정자 운동학적 특성과 Zygote arrest 1 유전자 변이와의 연관성 분석)

  • Lee, Mi Jin;Ko, Jun Ho;Kim, Yong Min;Choi, Tae Jeong;Cho, Kyu Ho;Kim, Young Sin;Jin, Dong Il;Kim, Nam Hyung;Cho, Eun Seok
    • ANNALS OF ANIMAL RESOURCE SCIENCES
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    • v.29 no.4
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    • pp.150-157
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    • 2018
  • The Zygote arrest 1 (ZAR1) gene is known to affect early embryonic development in various vertebrates. In this study, we performed the association analysis to check whether there is any significant relationship between semen kinematic characteristics and the ZAR1 gene. To determine semen kinematic characteristics, we measured motility (MOT), straight-line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), straightness (STR), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF) of spermatozoa in boars. In order to detect single nucleotide polymorphisms (SNPs), we extracted genomic DNA from multiple Duroc boars, and then subsequently used them in sequencing reactions. As a result, three SNPs were detected in the intronic region of ZAR1 gene (g.2435T>C in intron 2, g.2605G>A and g.4633A>C in intron 3 ). SNPs g.2435T>C and g.2605G>A were significantly associated with MOT (p<0.01) and VSL (p<0.05), and g.4633A

Epidemiology and Clinical Characteristics of Parainfluenza Virus Type 4 in Korean Children: a Single Center Study, 2015-2017 (소아에서 파라인플루엔자 바이러스 4형의 역학 및 임상 양상에 대한 단일기관 연구: 2015-2017)

  • Sohn, Young Joo;Choi, Youn Young;Yun, Ki Wook;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.25 no.3
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    • pp.156-164
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    • 2018
  • Purpose: We aimed to identify the epidemiology and the clinical characteristics of human parainfluenza virus type 4 (HPIV-4) infection compared to HPIVs 1-3 infections in Korean children. Methods: We reviewed medical records of children with HPIV infection who visited Seoul National University Children's Hospital from 2015 to 2017. Detection of respiratory viruses was performed using real time-polymerase chain reaction (rt-PCR), which could differentiate HPIVs 1-4. Diagnosis was classified as a febrile illness, upper respiratory tract infection (URI), croup, bronchiolitis, or pneumonia. The epidemiology, demographic features, and clinical characteristics among HPIV types were compared. The clinical data were analyzed only for the previously healthy children. Results: Of the 472 children diagnosed with HPIV infection, 108 (22.9%) were previously healthy: 24 (22.2%), 19 (17.6%), 39 (36.1%), and 26 (24.1%) in HPIV types 1, 2, 3, and 4, respectively. The median age of children with HPIV-4 infection was 11 (0-195) months: the proportion of children aged < 2 years and 2 to < 5 years were 65.4% and 19.2%, respectively. Clinical diagnoses of HPIV-4 infection were bronchiolitis (38.5%), pneumonia (30.8%), and URI (30.8%). Croup was the most prevalent in HPIV-2 (21.1%) and none in HPIV-4 infection (P=0.026). Hospital admission rates among HPIV types were not significantly different (P>0.05). Conclusions: We observed seasonal peak of HPIV-4 infection in 2015 and 2017. HPIV-4 was a common respiratory pathogen causing lower respiratory tract infection in hospitalized children.

Conditioned Media of RAW 264.7 Cells Stimulated with Phellinus linteus Extract Regulates the Epithelial-mesenchymal Transition in Prostate Cancer Cells (상황버섯에 의해 활성화된 RAW 264.7 대식세포주 배양액의 인간 전립선암 세포주의 epithelial-mesenchymal transition 조절)

  • Kang, Taewoo;An, Hyun-Hee;Park, Sul-Gi;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Ji-Won;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.29 no.8
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    • pp.904-915
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    • 2019
  • Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.