• 제목/요약/키워드: Real-Time Polymerase Chain Reaction

검색결과 795건 처리시간 0.033초

2009 H1N1 인플루엔자 폐렴에서 Procalcitonin의 유용성: 세균성 폐렴과의 감별 역할 (Procalcitonin in 2009 H1N1 Influenza Pneumonia: Role in Differentiating from Bacterial Pneumonia)

  • 안신;김원영;윤지영;손창환;서동우;김성한;홍상범;임채만;고윤석;김원
    • Tuberculosis and Respiratory Diseases
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    • 제68권4호
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    • pp.205-211
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    • 2010
  • Background: Procalcitonin is a well known marker in infection that plays a role in distinguishing between bacterial and viral infections in screening. The aim of the present study was to evaluate the role of procalcitonin in differentiating between 2009 H1N1 influenza pneumonia and community acquired pneumonia of bacterial origin, or mixed bacterial origin and 2009 H1N1 influenza infection. Methods: A retrospective observational study was performed over the 6-month winter period during the 2009 H1N1 influenza pandemic. Ninety-six patient-subjects were enrolled, all of whom had been diagnosed with community acquired pneumonia in emergency department during the study period. On admission, laboratory studies were performed, which included 2009 H1N1 influenza real-time polymerase chain reaction of nasal secretions and procalcitonin on serum; the laboratory values were compared between the study groups. Receiver operating characteristic curve analyses were performed on the resulting data. Results: Compared to those with bacterial or mixed infections (n=62) and bacterial pneumonia with confirmed organisms (n=30), patients with 2009 H1N1 pneumonia (n=34) were significantly more likely to have low procalcitonin levels (p=0.008, 0.001). Using cutoff of value >0.3 ng/mL, the sensitivity and specificity of procalcitonin for detection of patients with confirmed bacterial pneumonia were 76.2% and 60.6%, respectively. A significant difference in procalcitonin was found between 2009 H1N1 pneumonia and pneumonia caused by mixed influenza viral and bacterial infections (0.15 [0.05~0.84] vs. 10.3 [0.05~22.87] ng/mL, p=0.045). Conclusion: Serum procalcitonin measurement may assist in the discrimination between pneumonia of bacterial and of 2009 H1N1 influenza origin. High values of procalcitonin suggest that bacterial infection or mixed infection of bacteria and 2009 H1N1 influenza is more likely.

OMC-2010 추출물이 마우스의 비장세포 cytokine 생성에 미치는 영향 (Effects of OMC-2010 extracts on cytokine productions in mouse spleen cells)

  • 배기상;박경철;최선복;조일주;서상완;김종진;신용국;김민선;박규환;김현식;송호준;박성주
    • 대한본초학회지
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    • 제27권5호
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    • pp.21-25
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    • 2012
  • Objective : This study was performed to estimate the effects of OMC-2010 extract on cytokine production in mouse spleen cells. Methods : Mouse spleen cells were pre-treated with ethanol and water extract of OMC-2010 for 1 h, then stimulated with lipopolysaccharide (LPS, 1 ${\mu}g/ml$) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokines. Results : OMC-2010 ethanol extract significantly inhibited the LPS-induced interleukin (IL)-1beta, tumor necrosis factor (TNF)-alpha and IL-5 mRNA expressions, but not shown such changes in IL-6, IL-4, IL-13. OMC-2010 water extract significantly inhibited the LPS-induced TNF-alpha, and IL-5 mRNA expressions, but not shown such changes in IL-1beta, IL-6, IL-4, IL-13. Conclusions : Theses results could suggest that both ethanol and water OMC-2010 extract could inhibit the TNF-alpha and IL-5 mRNA expression.

Korean Red Ginseng attenuates anxiety-like behavior during ethanol withdrawal in rats

  • Zhao, ZhengLin;Kim, Young Woo;Wu, YiYan;Zhang, Jie;Lee, Ju-Hee;Li, XiaoHua;Cho, Il Je;Park, Sang Mi;Jung, Dae Hwa;Yang, Chae Ha;Kim, Sang Chan;Zhao, RongJie
    • Journal of Ginseng Research
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    • 제38권4호
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    • pp.256-263
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    • 2014
  • Background: Korean Red Ginseng (KRG) is known to have antianxiety properties. This study was conducted to investigate the anxiolytic effects of KRG extract (KRGE) during ethanol withdrawal (EW) and the involvement of the mesoamygdaloid dopamine (DA) system in it. Methods: Rats were treated with 3 g/kg/d of ethanol for 28 d, and subjected to 3 d of withdrawal. During EW, KRGE (20 mg/kg/d or 60 mg/kg/d, p.o.) was given to rats once/d for 3 d. Thirty min after the final dose of KRGE, anxiety-like behavior was evaluated in an elevated plus maze (EPM), and plasma corticosterone (CORT) levels were determined by a radioimmunoassay (RIA). In addition, concentrations of DA and 3,4-dihydroxyphenylacetic acid (DOPAC) in the central nucleus of the amygdala (CeA) were also measured by high performance liquid chromatography (HPLC). Results: The EPM test and RIA revealed KRGE inhibited anxiety-like behavior and the over secretion of plasma CORT during EW. Furthermore, the behavioral effect was blocked by a selective DA D2 receptor (D2R) antagonist (eticlopride) but not by a selective DA D1 receptor (D1R) antagonist (SCH23390). HPLC analyses showed KRGE reversed EW-induced decreases of DA and DOPAC in a dose-dependent way. Additionally, Western blotting and real-time polymerase chain reaction (PCR) assays showed that KRGE prevented the EW-induced reductions in tyrosine hydroxylase (TH) protein expression in the CeA and TH mRNA expression in the ventral tegmental area (VTA). Conclusion: These results suggest that KRGE has anxiolytic effects during EW by improving the mesoamygdaloid DA system.

성숙한 3T3-L1 지방세포에서 Akkermansia muciniphila의 지방축적 감소 효과 (Effect of Reduction in the Adipose Accumulation of Akkermansia muciniphila in Mature 3T3-L1 Adipocytes)

  • 심혜윤;임수경;신주현;이도경;서재구;김호준
    • 한방비만학회지
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    • 제19권2호
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    • pp.106-112
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    • 2019
  • Objectives: The aim of this study was to observe the reduction of lipid accumulation by treatment with Akkermansia muciniphila extract on 3T3-L1 adipocytes. Methods: After treating pasteurized Akk. muciniphila strains in HT-29 colorectal cancer cell, the relative expression of interleukin (IL)-8, tumor necrosis factor-α, IL-6, and IL-1β mRNA was analyzed by real time polymerase chain reaction, respectively. 27 strains of Akk. muciniphila which have anti-inflammatory effects were selected. 3T3-L1 pre-adipocytes were treated with Akk. muciniphila for 24 hr and then measured the toxicity using water soluble tetrazolium salt assay. The cells were incubated for 4 days and then differentiated into adipocytes using the medium including adipogenic reagents for 10 days. The Akk. muciniphila was treated when the medium was exchanged for differentiation medium at 4th day and insulin medium at 6th day. To observe the lipid accumulation, the cells were stained with Oil red O dye and were measured using a spectrophotometer. Results: In the cytotoxicity test, the cell viability of 3T3-L1 pre-adipocytes was significantly increased compared to the control group which untreated with Akk. muciniphila, and there was no cytotoxicity of Akk. muciniphila at 1×107 CFU/mL. The results on Oil red O staining and absorbance measurements were showed a significant decrease in lipid accumulation in the group which was treated with Akk. muciniphila compared to the control group. Conclusions: In our results, Akk. muciniphila has the inhibitory effect of lipid accumulation in 3T3-L1 adipocytes. This suggests that Akk. muciniphila could be help to improve obesity.

Evaluation of the in vivo skin moisturizing effects and underlying mechanisms of pomegranate concentrate solution and dried pomegranate concentrate powder

  • Kang, Su-Jin;Choi, Beom-Rak;Kim, Seung-Hee;Yi, Hae-Yeon;Park, Hye-Rim;Sung, Mi-Sun;Song, Chang-Hyun;Cho, Il-Je;Lee, Young-Joon;Ku, Sae-Kwang
    • 대한한의학회지
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    • 제37권2호
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    • pp.12-22
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    • 2016
  • Objectives: The aim of this study was to confirm the dose-dependent skin moisturizing effects of dried pomegranate concentrate powder (PCP) and pomegranate concentrate solution (PCS) in ICR mice. Materials and methods: To observe the in vivo skin moisturizing effects and possible underlying mechanisms of PCP and PCS, oral PCP (100, 200, and 400 mg/kg) and PCS (1, 2, and 4 mL/kg) were administered to normal ICR mice. Changes in body weight, skin water content, and skin type I collagen and hyaluronan contents were measured. Additionally, the mRNA expression levels of hyaluronan synthase (Has) 1, 2, and 3, and collagen type I alpha (COL1A) 1 and 2 were determined in the dorsal skin of mice by real-time reverse transcription polymerase chain reaction (RT-PCR). Results: Significant and dose-dependent increases in dorsal skin water content and type I collagen and hyaluronan contents were seen in PCP and PCS-treated mice. Moreover, the mRNA levels of Has 1, 2, and 3, involved in hyaluronan synthesis, and of COL1A1 and COL1A2, involved in collagen synthesis, were significantly and dose-dependently upregulated in PCS- and PCP-treated mice. Conclusions: In this study, PCP and PCS led to favorable skin moisturizing effects as indicated by increased skin water content and the upregulation of hyaluronan and collagen synthesis enzymes in mice treated with PCS (4 mL/kg) and PCP (200 mg/kg).

Vitamin D Inhibits Expression and Activity of Matrix Metalloproteinase in Human Lung Fibroblasts (HFL-1) Cells

  • Kim, Seo Hwa;Baek, Moon Seong;Yoon, Dong Sik;Park, Jong Seol;Yoon, Byoung Wook;Oh, Byoung Su;Park, Jinkyeong;Kim, Hui Jung
    • Tuberculosis and Respiratory Diseases
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    • 제77권2호
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    • pp.73-80
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    • 2014
  • Background: Low levels of serum vitamin D is associated with several lung diseases. The production and activation of matrix metalloproteinases (MMPs) may play an important role in the pathogenesis of emphysema. The aim of the current study therefore is to investigate if vitamin D modulates the expression and activation of MMP-2 and MMP-9 in human lung fibroblasts (HFL-1) cells. Methods: HFL-1 cells were cast into three-dimensional collagen gels and stimulated with or without interleukin-$1{\beta}$ (IL-$1{\beta}$) in the presence or absence of 100 nM 25-hydroxyvitamin D (25(OH)D) or 1,25-dihydroxyvitamin D ($1,25(OH)_2D$) for 48 hours. Trypsin was then added into the culture medium in order to activate MMPs. To investigate the activity of MMP-2 and MMP-9, gelatin zymography was performed. The expression of the tissue inhibitor of metalloproteinase (TIMP-1, TIMP-2) was measured by enzyme-linked immunosorbent assay. Expression of MMP-9 mRNA and TIMP-1, TIMP-2 mRNA was quantified by real time reverse transcription polymerase chain reaction. Results: IL-$1{\beta}$ significantly stimulated MMP-9 production and mRNA expression. Trypsin converted latent MMP-2 and MMP-9 into their active forms of MMP-2 (66 kDa) and MMP-9 (82 kDa) within 24 hours. This conversion was significantly inhibited by 25(OH)D (100 nM) and $1,25(OH)_2D$ (100 nM). The expression of MMP-9 mRNA was also significantly inhibited by 25(OH)D and $1,25(OH)_2D$. Conclusion: Vitamin D, 25(OH)D, and $1,25(OH)_2D$ play a role in regulating human lung fibroblast functions in wound repair and tissue remodeling through not only inhibiting IL-$1{\beta}$ stimulated MMP-9 production and conversion to its active form but also inhibiting IL-$1{\beta}$ inhibition on TIMP-1 and TIMP-2 production.

Elucidation of Bacterial Pneumonia-Causing Pathogens in Patients with Respiratory Viral Infection

  • Jung, Hwa Sik;Kang, Byung Ju;Ra, Seung Won;Seo, Kwang Won;Jegal, Yangjin;Jun, Jae-Bum;Jung, Jiwon;Jeong, Joseph;Jeon, Hee-Jeong;Ahn, Jae-Sung;Lee, Taehoon;Ahn, Jong Joon
    • Tuberculosis and Respiratory Diseases
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    • 제80권4호
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    • pp.358-367
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    • 2017
  • Background: Bacterial pneumonia occurring after respiratory viral infection is common. However, the predominant bacterial species causing pneumonia secondary to respiratory viral infections other than influenza remain unknown. The purpose of this study was to know whether the pathogens causing post-viral bacterial pneumonia vary according to the type of respiratory virus. Methods: Study subjects were 5,298 patients, who underwent multiplex real-time polymerase chain reaction for simultaneous detection of respiratory viruses, among who visited the emergency department or outpatient clinic with respiratory symptoms at Ulsan University Hospital between April 2013 and March 2016. The patients' medical records were retrospectively reviewed. Results: A total of 251 clinically significant bacteria were identified in 233 patients with post-viral bacterial pneumonia. Mycoplasma pneumoniae was the most frequent bacterium in patients aged <16 years, regardless of the preceding virus type (p=0.630). In patients aged ${\geq}16years$, the isolated bacteria varied according to the preceding virus type. The major results were as follows (p<0.001): pneumonia in patients with influenza virus (type A/B), rhinovirus, and human metapneumovirus infections was caused by similar bacteria, and the findings indicated that Staphylococcus aureus pneumonia was very common in these patients. In contrast, coronavirus, parainfluenza virus, and respiratory syncytial virus infections were associated with pneumonia caused by gram-negative bacteria. Conclusion: The pathogens causing post-viral bacterial pneumonia vary according to the type of preceding respiratory virus. This information could help in selecting empirical antibiotics in patients with post-viral pneumonia.

Molecular Cloning of the cDNA of Heat Shock Protein 88 Gene from the Entomopathogenic Fungus, Paecilomyces tenuipes Jocheon-1

  • Liu, Ya-Qi;Park, Nam Sook;Kim, Yong Gyun;Kim, Keun Ki;Park, Hyun Chul;Son, Hong Joo;Hong, Chang Ho;Lee, Sang Mong
    • International Journal of Industrial Entomology and Biomaterials
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    • 제28권2호
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    • pp.71-84
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    • 2014
  • The full-length heat shock protein 88 (HSP88) complementary DNA (cDNA) of Paecilomyces tenuipes Jocheon-1 was obtained by screening the Paecilomyces tenuipes (P. tenuipes) Jocheon-1 Uni-Zap cDNA library and performing 5' RACE polymerase chain reaction (PCR). The P. tenuipes Jocheon-1 HSP88 cDNA contained an open reading frame (ORF) of 2,139-basepair encoding 713 amino acid residues. The deduced amino acid sequence of the P. tenuipe s Jocheon-1 HSP88 cDNA showed 77% identity to Nectria haematococca HSP88 and 45-76% identity to other fungal homologous HSP88s. Phylogenetic analysis and BLAST program analysis confirmed that the deduced amino acid sequences of the P. tenuipes Jocheon-1 HSP88 gene belonged to the ascomycetes group within the fungal clade. The P. tenuipes Jocheon-1 HSP88 also contained the conserved ATPase domain at the N-terminal region. The cDNA encoding P. tenuipes Jocheon-1 HSP88 was expressed as an 88 kilodalton (kDa) polypeptide in baculovirus-infected insect Sf9 cells. Under higher temperature conditions for the growth of the entomopathogenic fungus, mRNA expression of P. tenuipes Jocheon-1 HSP88 was quantified by real time PCR (qPCR). The results showed that heat shock stress induced a higher level of mRNA expression compared to normal growth conditions.

Fermented ginseng extract, BST204, disturbs adipogenesis of mesenchymal stem cells through inhibition of S6 kinase 1 signaling

  • Yi, Sang Ah;Lee, Jieun;Park, Sun Kyu;Kim, Jeom Yong;Park, Jong Woo;Lee, Min Gyu;Nam, Ki Hong;Park, Jee Hun;Oh, Hwamok;Kim, Saetbyul;Han, Jihoon;Kim, Bo Kyung;Jo, Dong-Gyu;Han, Jeung-Whan
    • Journal of Ginseng Research
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    • 제44권1호
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    • pp.58-66
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    • 2020
  • Background: The biological and pharmacological effects of BST204, a fermented ginseng extract, have been reported in various disease conditions. However, its molecular action in metabolic disease remains poorly understood. In this study, we identified the antiadipogenic activity of BST204 resulting from its inhibition of the S6 kinase 1 (S6K1) signaling pathway. Methods: The inhibitory effects of BST204 on S6K1 signaling were investigated by immunoblot, nuclear fractionation, immunoprecipitation analyses. The antiadipogenic effect of BST204 was evaluated by measuring mRNA levels of adipogenic genes and by chromatin immunoprecipitation and quantitative real-time polymerase chain reaction analysis. Results: Treatment with BST204 inhibited activation and nuclear translocation of S6K1, further decreasing the interaction between S6K1 and histone H2B in 10T1/2 mesenchymal stem cells. Subsequently, phosphorylation of H2B at serine 36 (H2BS36p) by S6K1 was reduced by BST204, inducing an increase in the mRNA expression of Wnt6, Wnt10a, and Wnt10b, which disturbed adipogenic differentiation and promoted myogenic and early osteogenic gene expression. Consistently, BST204 treatment during adipogenic commitment suppressed the expression of adipogenic marker genes and lipid drop formation. Conclusion: Our results indicate that BST204 blocks adipogenesis of mesenchymal stem cells through the inhibition of S6K1-mediated histone phosphorylation. This study suggests the potential therapeutic strategy using BST204 to combat obesity and musculoskeletal diseases.

Hepatitis C Virus Nonstructural 5A Protein (HCV-NS5A) Inhibits Hepatocyte Apoptosis through the NF-κb/miR-503/bcl-2 Pathway

  • Xie, Zhengyuan;Xiao, Zhihua;Wang, Fenfen
    • Molecules and Cells
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    • 제40권3호
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    • pp.202-210
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    • 2017
  • The nonstructural protein 5A (NS5A) encoded by the human hepatitis C virus (HCV) RNA genome is a multifunctional phosphoprotein. To analyse the influence of NS5A on apoptosis, we established an Hep-NS5A cell line (HepG2 cells that stably express NS5A) and induced apoptosis using tumour necrosis factor $(TNF)-{\alpha}$. We utilised the MTT assay to detect cell viability, real-time quantitative polymerase chain reaction and Western blot to analyse gene and protein expression, and a luciferase reporter gene experiment to investigate the targeted regulatory relationship. Chromatin immunoprecipitation was used to identify the combination of $NF-{\kappa}B$ and miR-503. We found that overexpression of NS5A inhibited $TNF-{\alpha}$-induced hepatocellular apoptosis via regulating miR-503 expression. The cell viability of the $TNF-{\alpha}$ induced Hep-mock cells was significantly less than the viability of the $TNF-{\alpha}$ induced Hep-NS5A cells, which demonstrates that NS5A inhibited $TNF-{\alpha}$-induced HepG2 cell apoptosis. Under $TNF-{\alpha}$ treatment, miR-503 expression was decreased and cell viability and B-cell lymphoma 2 (bcl-2) expression were increased in the Hep-NS5A cells. Moreover, the luciferase reporter gene experiment verified that bcl-2 was a direct target of miR-503, NS5A inhibited $TNF{\alpha}$-induced $NF-{\kappa}B$ activation and $NF-{\kappa}B$ regulated miR-503 transcription by combining with the miR-503 promoter. After the Hep-NS5A cells were transfected with miR-503 mimics, the data indicated that the mimics could reverse $TNF-{\alpha}$-induced cell apoptosis and blc-2 expression. Collectively, our findings suggest a possible molecular mechanism that may contribute to HCV treatment in which NS5A inhibits $NF-{\kappa}B$ activation to decrease miR-503 expression and increase bcl-2 expression, which leads to a decrease in hepatocellular apoptosis.