• 대한임상검사과학회지
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• 제41권4호
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• pp.145-152
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• 2009

Novel influenza A virus, subtype H1N1 of swine-lineage, has been transmitted rapidly to many regions of the world. Rapid detection of the virus is essential to instigate appropriate patient care and public health management and for disease surveillance. The aim of this study is to determine the prevalence of novel influenza A (H1N1) virus in Korea using reverse-transcription real time polymerase chain reaction (rRT-PCR). Novel H1N1 virus was detected in a total of 8,948 nasopharyngeal samples from patients with influenza-like illness throughout Korea from August to September 2009. RNA was extracted from $300{\mu}l$of sample using an RNA extraction kit (Zymo Research, CA, USA). In the present study, Genekam kit (Genekam, Duisburg, Germany) was used to detect novel H1N1 virus. Novel H1N1 virus was found in 1,130 samples from a total of 8,948 samples (12.6%). The highest frequency was found in 10- to 19-year-olds (M: 29.3% vs. F: 16.4%), followed by 20- to 29-year-olds (M: 17.9% vs. F: 15.4%), 40- to 49-year-olds (M: 6.5% vs. F: 8.1%), 50- to 59-year-olds (M: 6.0% vs. F: 5.5%), and 30- to 39-year-olds (M: 4.6% vs. F: 3.8%). The mean positive rate was higher in men than in women (M: 14.7% vs. F: 7.4%). Novel H1N1 virus showed the lowest prevalence in patients over 60 years old. The positive rate increased daily and showed a significant high peak in mid-September 2009. In 19 provinces of Korea, Cheonan (41.1%), Busan (37.3%), Gangneung (33.3%), Jinju (32.1%), Ulsan (24.6%), Deajeon (23.7%) areas showed high frequencies and other provinces were found less than 10% of novel H1N1 virus. Since reverse-transcription real time PCR assay is rapid, accurate, and convenient, it may assist public health laboratories in detecting novel H1N1 virus. Moreover, these data could be useful for the management of patients with influenza-like illness.

• 대한임상검사과학회지
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• 제51권3호
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• pp.329-335
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• 2019

매년 호흡기 바이러스 감염으로 인해 수 백만명의 소아들이 사망한다. 호흡기 바이러스 감염의 원인 병원체 중 Human rhinovirus (HRV)는 코감기의 주요 원인 균으로 면역력이 약한 영, 유아, 노인 그리고 천식 환자에게 심각한 호흡기 감염의 원인으로 작용하는 병원체이다. 2012년 1월부터 2018년 12월까지 천안 단국대학교 병원 진단검사의학과에 호흡기 바이러스 검사가 의뢰된 호흡기 검체 9,010개의 검체를 real time reverse transcription PCR (real time RT-PCR) 방법으로 검사했다. 총 12종의 호흡기 바이러스를 real-time RT-PCR로 검출했다. 연구기간 중 평균 검출률은 21.3%이었고, HRV 양성 환자의 평균 연령은 6.5세였다. 7월의 검출률이 32.4%로 가장 높게 나타났고 2월이 8.3%로 가장 낮았다. 연령대별로 검출률을 분석해봤을 때 10세 미만의 검출률이 가장 높았다. HRV의 중복 감염률은 35.3%이고, 가장 흔한 조합은 Adenovirus와의 조합이었다. 호흡기 바이러스 감염증은 비슷한 임상 증상을 가지고 있어 빠른 진단이 이루어 져야 적절한 시기에 적절한 치료를 할 수 있다. 호흡기 바이러스 감염은 보통 면역력이 약한 어린아이와 노인에서 주로 발생하는 것으로 알려져 있다. 하지만 본 연구에서는 10세 미만에 이어 10대 환자들의 검출률이 높았다. 그리고 1,2월을 제외하고 15% 이상의 detection rate를 보였다. HRV의 감염 양상에 대한 꾸준한 연구가 필요할 것으로 사료된다.

• Fisheries and Aquatic Sciences
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• 제15권4호
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• pp.317-324
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• 2012

We characterized the cytoskeletal beta-actin (${\beta}$-ACT) gene (actb) and its 5'-upstream regulatory region in the Javanese ricefish Oryzias javanicus. The gene and protein structures were deduced from amino acid sequences of the actb gene and conserved in the teleost lineage. The O. javanicus actb gene has common transcription factor binding motifs in its regulatory region found in teleostean orthologues. Following quantitative reverse transcription-PCR, actb gene transcripts were detected in all tissues examined; however, the basal expression levels were different. During early development, O. javanicus actb mRNA levels showed a gradual increase and peaked between late somitogenesis and the heartbeat stage. Microinjection of O. javanicus embryos with the actb gene promoter-driven red fluorescent protein (RFP) gene reporter vector showed a ubiquitous distribution of RFP signals, although most exhibited a mosaic pattern of transgene expression. A small number of microinjected embryos displayed a wide distribution of RFP signals over their entire body, which resembled the expression pattern of endogenous actb. Data from this study provide a basis to develop a transgenic system with ubiquitous expression of foreign genes in O. javanicus.

• Journal of Microbiology
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• 제39권3호
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• pp.186-194
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• 2001

Scrub typhus, caused by Orientia tsutsugamushi infection, is clinically and histopathologically characterized by local as well as systemic inflammatory reactions, indicating that orientiae induce mechanisms that amplify the inflammatory response. To reveal underlying mechanisms of chemoattraction and activation of responding leukocytes, expression of chemokine and tumor necrosis factor alpha (TNF-$\alpha$) genes in murine peritoneal macrophages after infection with the obligate intracellular bacterium Ο.tsutsugamushi was investigated. The genes that were unregulated included macrophage inflammatory proteins l$\alpha$/$\beta$(MIP-l$\alpha$/$\beta$), MIP-2, monocyte chemoattractant protein 1(MCP-1), RANTES (regulated upon activation, normal T-cell expressed and secreted), gamma-interferon-inducible protein 10(IP-10) and TNF-$\alpha$. Peak expression of these chemokines and TNF-$\alpha$ was observed between 1 and 3 h after infection. These responses returned to or approached baseline preinfection levels 6 h after challenge. Semiquantitative reverse transcription (RT)-PCR analysis revealed dramatic Increases during infection in the steady-state levels of mRNA ceding for the inhibitory subunit of NF-kB (IkB$\alpha$), whose transcription is enhanced by binding of NF-kB within the IkB$\alpha$promoter region. Thus, Ο. tsutsugamushi appears to be a stung inducer of chemokines and TNF-$\alpha$ which may significantly contribute to inflammation and tissue damage observed in scrub typhus by attracting and activating phagocytic leukocytes.

• 동의생리병리학회지
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• 제34권2호
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• pp.67-74
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• 2020

The objective of this study is to investigate the skin whitening and antioxidant effects of the Anemarrhenae Rhizoma extract (ARE). Following the previously studied method, we examined the inhibitory effects of melanin synthesis and tyrosinase activity by using B16F10 cells. First, we measured the Diphenylpicrylhydrazyl (DPPH) assay, nitrite scavenging activity, and superoxide dismutase-like activity to verifying antioxidant efficacy according to skin whitening. In addition, we confirmed the skin whitening efficacy of ARE by measuring gene expression associated with a skin whitening by the Reverse transcription polymerase chain reaction (RT-PCR) method in B16F10 cells. In this study, we confirmed that ARE has skin whitening and antioxidant effects at high concentrations. In particular, ARE at a concentration of 500 ㎍/ml inhibited the expression of Tyrosinase, TRP-2 (tyrosinase-related protein), and MITF (microphthalmia transcription factor) genes better than Arbutin. In conclusion, our results confirmed that ARE has the potential for development as a skin whitening efficacy substance.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.303-310
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• 2011

We characterized a cytoplasmic actin gene locus in greenling Hexagrammos otakii (Scorpaeniformes). Genomic clones isolated from the greenling DNA library contained two homologous cytoplasmic actin gene copies (HObact2.1 and HObact2.2) in a tail-to-head orientation. Their gene structure is characterized by six translated exons and one non-translated exon. Exon-intron organization and the nucleotide sequences of the two actin gene isoforms are very similar. However, only the HObact2.1 isoform contains microsatellite-like, dinucleotide repeats in the 5'-flanking region (named HOms2002) and intron 1 following the non-translated exon 1 (named HOms769). One microsatellite locus (HOms769) was highly polymorphic while the other (HOms2002) was not. Based on bioinformatic analysis, different transcription factor binding motifs are related to stress and immune responses in the two actin isoforms. Semiquantitative and real-time reverse transcription-PCR assays showed that both isoform transcripts were detectable ubiquitously in all the tissues examined. However, the basal expression levels of each isoform varied across tissues. Overall, the two isoforms showed a similar, but not identical, expression pattern. Our data suggest that the cytoplasmic actin genes may be the result of a recent duplication event in the greenling genome, which has not experienced significant subfunctionalization in their housekeeping roles.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.836-843
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• 2004

Nontypeable H. influenzae (NTHi), a Gram-negative obligate human pathogen, causes pneumonia, chronic bronchitis, and otitis media, and the respiratory epithelium is the first line of defense that copes with the pathogen. In an effort to identify transcriptional responses of human respiratory epithelial cells to infection with NTHi, we examined its differential gene expression using high density cDNA microarrays. BEAS-2B human bronchial epithelial cells were exposed to NTHi for 3 hand 24 h, and the alteration of mRNA expression was analyzed using microarrays consisting of 8,170 human cDNA clones. The results indicated that approximately 2.6% of the genes present on the microarrays increased in expression over 2-fold and 3.8% of the genes decreased during the 24-h infection period. Upregulated genes included cytokines (granulocyte-macrophage colony stimulating factor 2, granulocyte chemotactic protein 2, IL-6, IL-10, IL-8), transcription factors (Kruppel-like factor 7, CCAAT/enhancer binding protein $\beta$, E2F-1, NF-$\kappa$B, cell surface molecules (CD74, ICAM-1, ICAM-2, HLA class I), as well as those involved in signal transduction and cellular transport. Selected genes were further confirmed by reverse-transcription-PCR. These data expand our knowledge of host cellular responses during NTHi infection and should provide a molecular basis for the study of host-NTHi interaction.

• Journal of Periodontal and Implant Science
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• 제35권3호
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• pp.799-811
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• 2005

This study was carried out to examine the potency of the three surface compo- nents from Porphyromonas gingivalis to stimulate the murine macrophage cell line RAW264.7 to synthesize the pro-inflammatory cytokine tumor necrosis factor alpha($TNF-{\alpha}$) and nitric oxide (NO). Lipopolysaccharide(LPS). lipid A-associated proteins(LAP) and saline-extractable surface -associated material(SAM) were isolated from P. gingivalis 381. $TNF-{\alpha}$ release into culture supernatants was determined by two-site ELISA. NO production was assayed by measuring the accumulation of nitrite in culture supernatants. Western blot analysis of iNOS and analysis of reverse transcription(RT)-PCR products were carried out. The surface components extracted from this bacterium were almost equally potent in stimulating release of $TNF-{\alpha}$ and NO by RAW264.7 cells. $TNF-{\alpha}$ that was being measured immunologically was due to activation of $TNF-{\alpha}$ gene transcription. The present study clearly shows that P. gingivalis surface components fully induced iNOS expression in RAW264.7 cells in the absence of other stimuli. The ability of P. gingivalis surface components to promote the production of $TNF-{\alpha}$ and NO may be important in the pathogenesis of inflammatory periodontal disease.

• Journal of Microbiology and Biotechnology
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• 제32권11호
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• pp.1406-1415
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• 2022

The formation of macrophage foam cells stimulated by oxidized low-density lipoprotein (ox-LDL) is deemed an important cause of atherosclerosis. Transcription factor Yin Yang 1 (YY1), which is a universally expressed multifunctional protein, is closely related to cell metabolism disorders such as lipid metabolism, sugar metabolism, and bile acid metabolism. However, whether YY1 is involved in macrophage inflammation and lipid accumulation still remains unknown. After mouse macrophage cell line RAW264.7 cells were induced by ox-LDL, YY1 and proprotein convertase subtilisin/kexin type 9 (PCSK9) expressions were found to be increased while low-density lipoprotein receptor (LDLR) expression was lowly expressed. Subsequently, through reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot analysis, Oil Red O staining and cholesterol quantification, it turned out that silencing of YY1 attenuated the inflammatory response and lipid accumulation in RAW264.7 cells caused by ox-LDL. Moreover, results from the JASPAR database, chromatin immunoprecipitation (ChIP) assay, luciferase reporter assay and Western blot analysis suggested that YY1 activated PCSK9 by binding to PCSK9 promoter and modulated the expression of LDLR in the downstream of PCSK9. In addition, the results of functional experiments demonstrated that the inhibitory effects of YY1 interference on ox-LDL-mediated macrophage inflammation and lipid accumulation were reversed by PCSK9 overexpression. To sum up, YY1 depletion inhibited its activation of PCSK9, thereby reducing cellular inflammatory response, cholesterol homeostasis imbalance, and lipid accumulation caused by ox-LDL.