• Title/Summary/Keyword: 실시간 정량적 중합효소연쇄반응

Search Result 19, Processing Time 0.024 seconds

Comparison of Gene Expression from Supernumerary Dental Pulp and Periodontal Ligament Stem Cells (과잉치 치수 세포와 치주인대 세포의 유전자 발현 비교)

  • Lee, Sangeun;Kim, Jongbin;Kim, Jongsoo
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.45 no.2
    • /
    • pp.242-249
    • /
    • 2018
  • The purpose of this study is to compare the properties of dental pulp and periodontal ligament stem cells from extracted supernumerary teeth by quantitative real-time PCR. Impacted supernumerary teeth in the maxillary anterior region were extracted. Dental pulp and periodontal ligament cells were collected from extracted supernumerary teeth on the same day. After isolation and culture of cells, compare characterization of them by using qRT-PCR. Primer sequences for odontoblasts are ONT, ALP, OCN, DMP-1 and DSPP. On dental pulp group, ONT has the largest quantity of gene expression, followed by OCN, ALP, DMP-1 and DSPP. On periodontal ligament group, ONT has the largest quantity of gene expression, followed by OCN, ALP, DSPP and DMP-1. Analysis of quantitative gene expression data using relative quantification showed that the expression of all genes decreased in periodontal ligament cells. Dental pulp and periodontal ligament stem cells from supernumerary teeth have the properties of odontoblasts. Considering that properties, supernumerary teeth were considered a useful donor site of dental pulp and periodontal ligament stem cells.

Anti-inflammatory Effect of Ishige foliacea in RAW 264.7 Cells (넓패추출물에 의한 RAW 264.7 세포에서의 항염효과)

  • Joonghyun Shim
    • Journal of the Society of Cosmetic Scientists of Korea
    • /
    • v.50 no.1
    • /
    • pp.29-36
    • /
    • 2024
  • This study was carried out to identify the anti-inflammatory effects of Ishige foliacea (I. foliacea) extract on skin using RAW 264.7 cells. The anti-inflammatory effects of I. foliacea extract on RAW 264.7 cells were assessed by cell viability assay, mRNA expressions, and nitric oxide (NO)/prostaglandin E2 (PGE2) productions. The anti-inflammatory effects of I. foliacea extract were elucidated by analysis of IL-1α/IL-1β/IL-6/TNFα gene expressions and PGE2/NO production. Quantitative real-time polymerase chain reaction showed that I. foliacea extract decreased the gene expression levels of iNOS/COX2/IL-1α/IL-1β and IL-6. Furthermore, PGE2/NO production also revealed that I. foliacea extract exhibited anti-inflammatory properties. These results suggest that I. foliacea extract is an anti-inflammatory compound. It could be a potent cosmeceutical material for anti-inflammatory effects. Further studies on the anti-inflammatory mechanisms of broadleaf extracts are expected to help identify pharmacological mechanisms related to inflammation in addition to cosmeceuticals.

Analysis of HBeAg and HBV DNA Detection in Hepatitis B Patients Treated with Antiviral Therapy (항 바이러스 치료중인 B형 간염환자에서 HBeAg 및 HBV DNA 검출에 관한 분석)

  • Cheon, Jun Hong;Chae, Hong Ju;Park, Mi Sun;Lim, Soo Yeon;Yoo, Seon Hee;Lee, Sun Ho
    • The Korean Journal of Nuclear Medicine Technology
    • /
    • v.23 no.1
    • /
    • pp.35-39
    • /
    • 2019
  • Purpose Hepatitis B virus (hepatitis B virus, HBV) infection is a worldwide major public health problem and it is known as a major cause of chronic hepatitis, liver cirrhosis and liver cancer. And serologic tests of hepatitis B virus is essential for diagnosing and treating these diseases. In addition, with the development of molecular diagnostics, the detection of HBV DNA in serum diagnoses HBV infection and is recognized as an important indicator for the antiviral agent treatment response assessment. We performed HBeAg assay using Immunoradiometric assay (IRMA) and Chemiluminescent Microparticle Immunoassay (CMIA) in hepatitis B patients treated with antiviral agents. The detection rate of HBV DNA in serum was measured and compared by RT-PCR (Real Time - Polymerase Chain Reaction) method Materials and Methods HBeAg serum examination and HBV DNA quantification test were conducted on 270 hepatitis B patients undergoing anti-virus treatment after diagnosis of hepatitis B virus infection. Two serologic tests (IRMA, CMIA) with different detection principles were applied for the HBeAg serum test. Serum HBV DNA was quantitatively measured by real-time polymerase chain reaction (RT-PCR) using the Abbott m2000 System. Results The detection rate of HBeAg was 24.1% (65/270) for IRMA and 82.2% (222/270) for CMIA. Detection rate of serum HBV DNA by real-time RT-PCR is 29.3% (79/270). The measured amount of serum HBV DNA concentration is $4.8{\times}10^7{\pm}1.9{\times}10^8IU/mL$($mean{\pm}SD$). The minimum value is 16IU/mL, the maximum value is $1.0{\times}10^9IU/mL$, and the reference value for quantitative detection limit is 15IU/mL. The detection rates and concentrations of HBV DNA by group according to the results of HBeAg serological (IRMA, CMIA)tests were as follows. 1) Group I (IRMA negative, CMIA positive, N = 169), HBV DNA detection rate of 17.7% (30/169), $6.8{\times}10^5{\pm}1.9{\times}10^6IU/mL$ 2) Group II (IRMA positive, CMIA positive, N = 53), HBV DNA detection rate 62.3% (33/53), $1.1{\times}10^8{\pm}2.8{\times}10^8IU/mL$ 3) Group III (IRMA negative, CMIA negative, N = 36), HBV DNA detection rate 36.1% (13/36), $3.0{\times}10^5{\pm}1.1{\times}10^6IU/mL$ 4) Group IV(IRMA positive, CMIA negative, N = 12), HBV DNA detection rate 25% (3/12), $1.3{\times}10^3{\pm}1.1{\times}10^3IU/mL$ Conclusion HBeAg detection rate according to the serological test showed a large difference. This difference is considered for a number of reasons such as characteristics of the Ab used for assay kit and epitope, HBV of genotype. Detection rate and the concentration of the group-specific HBV DNA classified serologic results confirmed the high detection rate and the concentration in Group II (IRMA-positive, CMIA positive, N = 53).

Validity of the New Caries Activity Test using Real-Time Polymerase Chain Reaction (실시간 중합효소연쇄반응 방법을 이용한 새로운 치아우식 활성 검사법의 유효성)

  • Kwon, Doyoun;Kim, Heejin;Nam, Okhyung;Kim, Misun;Choi, Sungchul;Kim, Kwangchul;Lee, Hyoseol
    • Journal of the korean academy of Pediatric Dentistry
    • /
    • v.45 no.3
    • /
    • pp.354-362
    • /
    • 2018
  • Periogen is a new caries activity test using real-time polymerase chain reaction. The aim of this study was to assess the validity of Periogen by evaluating the correlation with dmft, dmfts indices and comparing with Cariview and caries risk assessment tool (CAT). 83 children under 6 participated in this study. Dmft, dmfts indices and CAT were collected through an examination of oral health status. Plaque samples for Periogen and Cariview were collected and manipulated according to the manufactures' instructions. The correlation coefficient of Periogen, Cariview and CAT with the dmfts index were 0.38, 0.56 and 0.66 in each (p < 0.01). The sensitivity of Periogen, Cariview and CAT were 43%, 76% and 95% and specificity were 80%, 72% and 74% respectively. Area under curve under the receiver operating characteristic curves in each method indicated 0.69, 0.81 and 0.85. CAT and Cariview were more effective in evaluation the risk of dental caries than Periogen so far. To be used Periogen clinically, more improvements for higher validity were needed.

Comparison of Quantitative Relationship between Real-Time PCR and Acid Fast Bacilli Staining for Diagnosis of Pulmonary Tuberculosis (폐결핵 진단을 위한 실시간중합효소연쇄반응과 AFB 염색진단검사의 정량적 연관성 비교)

  • Jung, Taewon;Kim, Sang-Ha;Kim, Sunghyun;Choi, Jae-Sun;Kim, Young-Kwon
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.52 no.4
    • /
    • pp.335-341
    • /
    • 2020
  • This study investigates the association of the AFB stain with the cycle threshold (Ct) value of the Cobas TaqMan MTB test (CTM test, Roche Diagnostics, Basel, Switzerland), and it establishes the base data for semi-quantitative identification of M. tuberculosis by the Ct value. CTM test were simultaneously conducted on 8,389 specimens submitted to the Samsung Medical Center from January 2015 to December 2015, and the results were analyzed and compared retrospectively investigates the association of the AFB stain with the Ct value of the CTM test, and it establishes the base data for semi-quantitative identification of M. tuberculosis by the Ct value. The Ct values for 135 positive specimens of the CTM were inversely correlated with the AFB stain (rs=-0.545, P<0.01). When the Ct value of the CTM test and the time to positivity (TTP) of the mycobacteria cultures were verified based on the AFB stain, they were found to have a positive correlation (rs=0.136, P<0.01). The negative correlation between the CTM test and the AFB stain grade was demonstrated. The clinical significance was verified by applying these criteria to the clinical results. The semi-quantitative criteria of this study can be used to facilitate the rapid isolation of patients with active tuberculosis and infection control in the hospital.

Mitochondrial DNA Copy Number in the Patients of Korean Polycystic Ovary Syndrome (PCOS) (한국인 다낭성난소증후군 환자에서 미토콘드리아 DNA Copy 수의 정량적 분석)

  • Park, Ji-Eun;Jang, Min-Hee;Cho, Sung-Won;Kim, Yoo-Shin;Won, Hyung-Jae;Cho, Jung-Hyun;Baek, Kwang-Hyun;Lee, Sook-Hwan
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.33 no.4
    • /
    • pp.245-251
    • /
    • 2006
  • Objective: We analyzed quantification of mitochondria DNA (mtDNA) to investigate the relationship of mitochondria and pathogenesis of PCOS. Materials and Methods: Peripheral blood samples were collected from 28 patients with PCOS who were under the inclusion criteria for PCOS and from 28 healthy controls. Genomic DNA was used to analyze real-time PCR for mtDNA copy number quantification. The mtDNA copy number was compared between the control and PCOS groups. All data was expressed as mean ${\pm}$ SD. Statistical analysis was assessed by t-test. Results: In this study, the mtDNA $C_T$ was $11.67{\pm}0.422$ in PCOS patients and $11.51{\pm}0.722$ in control group, respectively. The mtDNA copy number was $1726410.71{\pm}407858.591$ the patients of in PCOS and $2167887.51{\pm}252459.28$ in control group (p=0.08), respectively. Conclusion: In our study, using real-time PCR, there was a tendency of lower mtDNA copy number in the patients of PCOS when comparing to the control group even though statistical difference was not significant. However, more extensive analysis is required to clarity relationship between mtDNA copy number and pathogenesis of PCOS.

Analysis of Stress Response of Domestic Chicken Breeds for the Development of a New Synthetic Parent Stock (국산 종계 개발을 위한 토종 계통들의 스트레스 반응 정도 분석)

  • Sohn, Sea Hwan;Cho, Eun Jung;Park, Ji Ae;Hong, Young Ho;Kim, Chong Dae
    • Korean Journal of Poultry Science
    • /
    • v.42 no.2
    • /
    • pp.157-167
    • /
    • 2015
  • We compared the degrees of stress response of 12 domestic purebred chicken strains that have been bred at National Institute of Animal Science, RDA, Korea since 1980. As a physiological marker of stress response, the expression levels of heat shock protein (HSP)-70, HSP-$90{\alpha}$, HSP-$90{\beta}$, hydroxyl-3-methyl-glutaryl coenzyme A reductase (HMGCR) genes and telomere length were measured by quantitative real-time polymerase chain reaction using the lymphocytes of 1,101 chickens. There was significant difference in HSP-70, HSP-$90{\alpha}$, HMGCR expression and telomere length among the strains. There was also significant difference in HSP-$90{\alpha}$, HSP-$90{\beta}$, and HMGCR expression between male and female chickens. Different age groups of chicken exhibited different expression levels of HSP-70, HSP-$90{\alpha}$ and telomere length. The results of the HSPs expression level suggested that, the strains of R, L and Y were highly resistant to stress, whereas the strains of S, O and W were susceptible to stress. Although the statistical differences in some of HSPs gene expression existed between genders, the HSP expression results varied in different strains that some opposed to the others, and there might be interaction between strains and genders, which conclude that there was no difference in stress response between male and female chickens. Moreover, despite of significant difference in some of HSPs expression level, it was considered that there was no difference in stress response between ages due to the inconsistent trends among HSP markers.

TNF-α stimulated IL-8 and IL-10 expression in monocytes from patients with chronic granulomatous disease (만성육아종질환 환자 단핵구에서 TNF-α 자극에 의한 IL-8과 IL-10의 발현 양상)

  • Shin, Kyung-Sue
    • Clinical and Experimental Pediatrics
    • /
    • v.51 no.10
    • /
    • pp.1096-1101
    • /
    • 2008
  • Purpose : Patients with chronic granulomatous disease (CGD) have genetic mutations in a component of the NADPH oxidase enzyme that is necessary for the generation of the superoxide anion. The profound defect in innate immunity is reflected by the patients susceptibility to catalase-positive bacteria and fungi. In addition, CGD patients display signs of persistent inflammation, which is not associated only with deficient superoxide anion production. The aim of this study was to elucidate the cytokine responses in CGD patients after $TNF-{\alpha}$ stimulation. Methods : Heparinized blood samples were collected from 8 CGD patients and 10 healthy volunteers. Monocytes ($1{\times}10^6cell/well$) isolated by the magnet cell isolation system were incubated with a constant amount of $TNF-{\alpha}$ (10 ng/mL) at $37^{\circ}C$ for 6 h. Incubated cells were harvested at 60-min intervals for IL-8 and IL-10 mRNA analysis, and the supernatant was collected at the same intervals to determine IL-8 and IL-10 expression. Monocytes from healthy volunteers were also incubated with antioxidants followed by $TNF-{\alpha}$ stimulation for IL-8 and IL-10 expression. Results : In CGD patients, a high expression of IL-8 together with a significantly higher IL-10 expression than in the healthy controls was seen after $TNF-{\alpha}$ stimulation. Moreover, normal monocytes treated with antioxidants exhibited increased IL-8 responses. Conclusion : The absence of phagocyte-derived reactive oxidants in CGD might be associated with a dysregulated production of pro- and antiinflammatory cytokines. Additional research related to reactive oxidants is needed to clarify the role of cytokines in CGD patients.

Resveratrol Ameliorates NMDA-induced Mitochondrial Injury by Enhanced Expression of Heme Oxygenase-1 in HT-22 Neuronal Cells (NMDA를 처리한 HT-22 신경세포에서 미토콘드리아 손상을 완화하는 레스베라트롤의 보호 효과와 헴 산화효소-1의 역할)

  • Kang, Jae Hoon;Woo, Jae Suk
    • Journal of Life Science
    • /
    • v.32 no.1
    • /
    • pp.11-22
    • /
    • 2022
  • N-methyl-D-aspartate (NMDA) receptors have received considerable attention regarding their involvement in glutamate-induced neuronal excitotoxicity. Resveratrol has been shown to exhibit neuroprotective effects against this kind of overactivation, but the underlying cellular mechanisms are not yet clearly understood. In this study, HT-22 neuronal cells were treated with NMDA in Mg2+-free buffer and subsequently used as an experimental model of glutamate excitotoxicity to elucidate the mechanisms of resveratrol-induced neuroprotection. We found that NMDA treatment causes a drop in MTT reduction ability, disrupts inside-negative transmembrane potential of mitochondria, depletes cellular ATP levels, and stimulates intracellular ROS production. Double fluorescence imaging studies demonstrated an increased formation of mitochondrial permeability transition (MPT) pores accompanied by apoptotic cell death, while cobalt protoporphyrin and bilirubin showed protective effects against NMDA-induced mitochondrial injury. On the other hand, zinc protoporphyrin IX significantly attenuated the protective effects of resveratrol which was itself shown to enhance heme oxygenase-1 (HO-1) mRNA and protein expression levels. In cells transfected with HO-1 small interfering RNA, resveratrol failed to suppress the NMDA-induced effects on MTT reduction ability and MPT pore formation. The present study suggests that resveratrol may prevent mitochondrial injury in NMDA- treated HT-22 cells and that enhanced expression of HO-1 is involved in the underlying cellular mechanism.