• Title/Summary/Keyword: 분자 진단

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Tissue Distribution of HuR Protein in Crohn's Disease and IBD Experimental Model (염증성 장질환 모델 및 크론병 환자에서의 점막상피 HuR 단백질의 변화 분석)

  • Choi, Hye Jin;Park, Jae-Hong;Park, Jiyeon;Kim, Juil;Park, Seong-Hwan;Oh, Chang Gyu;Do, Kee Hun;Song, Bo Gyoung;Lee, Seung Joon;Moon, Yuseok
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1339-1344
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    • 2014
  • Inflammatory bowel disease is an immune disorder associated with chronic mucosal inflammation and severe ulceration in the gastrointestinal tract. Antibodies against proinflammatory cytokines, including TNF${\alpha}$, are currently used as promising therapeutic agents against the disease. Stabilization of the transcript is a crucial post-transcriptional process in the expression of proinflammatory cytokines. In the present study, we assessed the expression and histological distribution of the HuR protein, an important transcript stabilizer, in tissues from experimental animals and patients with Crohn's disease. The total and cytosolic levels of the HuR protein were enhanced in the intestinal epithelia from dextran sodium sulfate (DSS)-treated mice compared to those in control tissues from normal mice. Moreover, the expression of HuR was very high only in the mucosal and glandular epithelium, and the relative localization of the protein was sequestered in the lower parts of the villus during the DSS insult. The expression of HuR was significantly higher in mucosal lesions than in normal-looking areas. Consistent with the data from the animal model, the expression of HuR was confined to the mucosal and glandular epithelium. These results suggest that HuR may contribute to the post-transcriptional regulation of proinflammatory genes during early mucosal insults. More mechanistic investigations are warranted to determine the potential use of HuR as a predictive biomarker or a promising target against IBD.

Development of Broad-range and Specific 16S rRNA PCR for Use in Routine Diagnostic Clinical Microbiology (임상미생물 검출을 위한 광대한 범위와 특이도를 가지는 16S rRNA PCR법 개발)

  • Kim, Hyun-Chul;Kim, Yun-Tae;Kim, Hyogyeong;Lee, Sanghoo;Lee, Kyoung-Ryul;Kim, Young-Jin
    • Journal of Life Science
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    • v.24 no.4
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    • pp.361-369
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    • 2014
  • Broad-range and specific 16S rRNA gene PCR is used for detection and identification of bacterial pathogens in clinical specimens from patients with a high suspicion for infection. We describe the development of a broad-range and specific PCR primer, based on bacterial 16S rRNA, for use in routine diagnostic clinical microbiology services. The primers were designed by using conservative regions of 16S rRNA sequences from 10 strains. Ninety-eight clinical strains were isolated from clinical patient specimens. A total of 98 strains of bacteria were identified by phenotypic methods; PCR with newly designed primers and universal primers. All purified PCR products were sequenced using both forward and reverse primers on an automated DNA analyzer. In this study, we evaluated the usefulness of the newly designed primers and the universal primers for the detection of bacteria, and both these techniques were compared with phenotypic methods for bacteria detection. When we also tested 98 strains of clinical isolates with newly designed primers, about 778 bp DNA fragments were amplified and identified from all strains. Of the 98 strains, 94 strains (95.9%) correspond in comparison with phenotypic methods. The newly designed primers showed that the identities of 98 (100%) strains were the same as those obtained by universal PCR primers. The overall agreement between the newly designed primers and universal primers was 100%. The primer set was designed for rapid, accurate, and cheap identification of bacterial pathogens. We think the newly designed primer set is useful for the identification of pathogenic bacteria.

Mutation Patterns of gyrA, gyrB, parC and parE Genes Related to Fluoroquinolone Resistance in Ureaplasma Species Isolated from Urogenital Specimens (비뇨생식기계 검체로부터 분리된 Ureaplasma 종의 Fluoroquinolone 내성과 관련된 gyrA, gyrB, parC, parE 유전자의 돌연변이 양상)

  • Cho, Eun-Jung;Hwang, Yu Yean;Koo, Bon-Kyeong;Park, Jesoep;Kim, Young Kwon;Kim, Sunghyun
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.2
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    • pp.74-81
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    • 2016
  • Ureaplasma species can normally colonize in the bodies of healthy individuals. Their colonization is associated with various diseases including non-gonococcal urethritis, chorioamnionitis, neonatal meningitis, and prematurity. In 2012, the sum of the resistant and intermediate resistant rates of Ureaplasma spp. to ofloxacin and ciprofloxacin was 66.08% and 92.69%, respectively. DNA point mutations in the genes encoding DNA gyrase (topoisomerase II) and topoisomerase IV are commonly responsible for fluoroquinolone resistance. Each enzyme is composed of two subunits encoded by gyrA and gyrB genes for DNA gyrase and parC and parE genes for topoisomerase IV. In the current study, these genes were sequenced in order to determine the role of amino acid substitutions in Ureaplasma spp. clinical isolates. From December 2012 to May 2013, we examined mutation patterns of the quinolone resistance-determining region (QRDR) in Ureaplasma spp. DNA sequences in the QRDR region of Ureaplasma clinical isolates were compared with those of reference strains including U. urealyticum serovar 8 (ATCC 27618) and U. parvum serovar 3 (ATCC 27815). Mutations were detected in all ofloxacin- and ciprofloxacin-resistant isolates, however no mutations were detected in drug-susceptible isolates. Most of the mutations related to fluoroquinolone resistance occurred in the parC gene, causing amino acid substitutions. Newly found amino acid substitutions in this study were Asn481Ser in GyrB; Phe149Leu, Asp150Met, Asp151Ile, and Ser152Val in ParC; and Pro446Ser and Arg448Lys in ParE. Continuous monitoring and accumulation of mutation data in fluoroquinolone-resistant Ureaplasma clinical isolates are essential to determining the tendency and to understanding the mechanisms underlying antimicrobial resistance.

The Usefulness Assessment of Attenuation Correction and Location Information in SPECT/CT (SPECT/CT에서 감쇠 보정 및 위치 정보의 유용성 평가)

  • Choi, Jong-Sook;Jung, Woo-Young;Shin, Sang-Ki;Cho, Shee-Man
    • The Korean Journal of Nuclear Medicine Technology
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    • v.12 no.3
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    • pp.214-221
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    • 2008
  • Purpose: We make a qualitative analysis of whether Fusion SPECT/CT can find lesion's anatomical sites better than existing SPECT or not, and we want to show the usefulness of SPECT/CT through finding out effects of CT attenuation correction on SPECT images. Materials and Method: 1. The evaluation of fusion images: This study comprised patients who was tested $^{131}I$-MIBG, Bone, $^{111}In$-Octreotide, Meckel's diverticulum, Parathyroid MIBI with Precedence 16 or Symbia T2 from 2008 Jan to Aug. We compared SPECT/CT image with non fusion image and make a qualitative analysis. 2. The evaluation of attenuation correction: We classified 38 patients who was tested 201Tl myocardial exam with Symbia T2 into 5 sections by using Cedars Sinai' QPS program - Ant, Inf, Lat, Septum, Apex. And we showed each section's perfusion states by percentage. We compared the each section's perfusion-states differences between CT AC and Non AC by average${\pm}$standard deviation. Results: 1. The evaluation of fusion images : In high energy $^{131}I$ cases, it was hard to grasp exact anatomical lesions due to difference between regions and surrounding lesions' uptake level. After combining with CT, we could grabs anatomical lesion more exactly. And in meckel's diverticulum case or to find lesions around bowels or organs with $^{111}In$ cases, it demonstrates its superiority. Bone SPECT/CT images help to distinguish between disk spaces certainly and give correct results. 2. The evaluation of attenuation correction: There is no significant difference statistically in Ant and Lat (p>0.05), but there is a meaningful difference in Inferior, Apex and Septum (p<0.05). AC perfusion at inferior wall in the 5 sections of myocardium: The perfusion difference between Non AC perfusion image ($68.58{\pm}7.55$) and CT corrected perfusion image ($76.84{\pm}6.52$) was the largest by $8.26{\pm}4.95$ (p<0.01, t=10.29). Conclusion: Nuclear medicine physicians can identify not only molecular image which shows functional activity of lesions but also anatomical location information of lesions with more accuracy using the combination of SPECT and CT systems. Of course this combination helps nuclear medicine physician find out the abnormal parts. Moreover combined data sets help separate between normal group and abnormal group in complicated body part. So clinicians can carry out diagnosis and treatment planning at the same time with a single test image. In addition, when we examine a myocardium in thorax where attenuation can occur easily, we can trust perfusion more in a certain region in SPECT test because CT provides the capability for accurate attenuation correction. In these reasons, we think we can prove the justice after treatment fusion image.

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Detection of Lily symptomless virus, Lily mottle virus, and Cucumber mosaic virus from Lilium Grown in Korea by RT-PCR (RT-PCR법을 이용한 백합 바이러스 LSV, LMoV, CMV의 검출)

  • Lim, Ji-Hyun;Bae, Eun-Hye;Lee, Yong-Jin;Park, Sung-Han;Lee, Kyu-Jun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.251-256
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    • 2009
  • Leaf samples and bulbs showing characteristic symptoms of virus infection were collected from Gang-won, Chung-nam, and Jeju Province of Korea in 2008-2009. Three viruses, Lily symptomless virus (LSV), Lily mottle virus (LMoV), and Cucumber mosaic virus (CMV) were detected by RT-PCR. Virus-infected plant samples were identified; 12 plants with LSV, 20 plants with LMoV, and 1 plant with CMV. Of the twelve LSV infected samples, seven samples were found to be mix-infected with LMoV and LSV. Symptoms of LMoV and LSV mixed infection were fairly severe, like as vein clearing, leaf curling, leaf mottling, leaf mosaic, and yellow streaking. Mixed infection with LMoV and LSV was also found in lily bulbs which have been stored under unfavorable environmental conditions. LMoV predominated in our tests, whereas spread of Lilyvirus X (LVX) was not found. The nucleotide sequences of coat protein (CP) region of seven isolates (4 LMoV, 2 LSV, and 1 CMV) were compared with the corresponding regions of LMoV (AJ564636), LSV (AJ516059) and CMV(AJ296154). The nucleotide sequence homologies between reference viruses and seven isolates were 95-99%. Complete sequencing of seven isolates is necessary to obtain more information on the molecular characteristics of these viruses as well as to increase sensitivity and rapidity of viral detection.

Genetic diversity of Clostridium perfringens form food-poisoning outbreak in Gyeonggi-do, 2013-2014 (경기도 식중독에서 분리된 Clostridium perfringens의 유전적 특성 분석)

  • Park, Sung-Hee;Choi, Ok-Kyung;Jeong, Jin-A;Kim, Woon-Ho;Lee, Yea-Eun;Park, Kwang-Hee;Yoon, Mi-Hye
    • Korean Journal of Microbiology
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    • v.52 no.3
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    • pp.286-297
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    • 2016
  • Clostridium perfringens is both a ubiquitous environmental bacterium and a major cause of human gastrointestinal disease, and C. perfringens food poisoning ranks among the most common gastrointestinal diseases in developed countries. 120 isolates of C. perfringens were obtained from food-poisoning outbreaks in 2013~2014, Gyeonggi-do. Using PCR, all 120 isolates were identified as C. perfringens type A. Of the tested isolates, 49 isolates carried the cpe gene, 71 isolates carried the cpb2 gene. The outbreak cases of cpb2 and cpe /cpb2 genes were 7 and 7, whereas the outbreak cases of cpe-gene were 2. The epidemiological relationship between C. perfringens isolates has previously been investigated chiefly by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST). The genetics relatedness of the isolates raged from 53.5-100% and 75 district PFGE type were observed. The PFGE results revealed a wide genetic diversity among the 64 cpb2 carrying isolates (except 7 isolates), while 46 cpe-carrying isolates (except 3 isolates) showed a high genetic similarity. The MLST analysis revealed that 14 cpe isolates (cpe-chromosomal isolates) belong to a distinct cluster that is significantly distant from all the other cpb2 isolates (cpe-plasmid carrying and cpe-negative isolates). The isolates carrying a cpb2 appear to be rarely related, and are more variable than chromosomal cpe isolates. The results suggest that the cpe-positive outbreak isolates showed close genetic relation, whereas the cpb2-positive isolates revealed a wide genetic diversity.

Correlation between Clinicopathology and Expression of HSP70, BAG1 and Raf-1 in Human Diffuse Type Gastric Carcinoma (미만형 위암에서 임상병리학적 인자와 Hsp70, BAG1과 Raf-1 발현간의 상관성)

  • Jung, Sang Bong;Lee, Hyoun Wook;Chung, Kyung Tae
    • Journal of Life Science
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    • v.26 no.1
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    • pp.101-108
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    • 2016
  • The aim of this study was to evaluate the relationships between the expression of Heat shock protein70 (HSP70), Raf-1 and Bcl-2-associated athanogene-1 (BAG1) protein in diffuse type gastric carcinoma and examine association of HSP70, Raf-1 and BAG1 expression with various clinic-pathological factors and survival. Heat shock protein70 is induced in the cells in response to various stress conditions, including carcinogens. Overexpression of heat shock protein 70 has been observed in many types of cancer. The proto-oncoprotein Raf is pivotal for mitogen-activated protein kinase (MAPK) signaling, and its aberrant activation has been implicated in multiple human cancers. Overexpression of BAG1 protein has been documented in some type of human cancer. BAG1 has been reported to interact with protein involved with a variety of signal pathway, and regulation of cell differentiation, survival and apoptosis. These interaction partners include HSP70 and Raf-1. The percentage of tumors exhibiting HSP70 positivity was significantly in cases of positive lymph node metastasis (64.9%) compared to cases without lymph node metastasis (35.1%, p=0.007). HS70 expression was correlated with pathological N-stage (p=0.006). Expression of BAG1 was detected in the majority of diffuse type gastric carcinoma tissues (71.7%), especially in younger patients (80% vs 52.6%, p=0.035). Furthermore BAG1 expression was correlated with tumor size (p=0.020). Raf-1 expression was found to be significantly associated with tumor size (p=0.005). The result indicate that HSP70 was significantly correlated the progression of diffuse type gastric cancer. Expression of BAG1 and Raf-1 may be used as diagnostic markers for gastric carcinoma.

Polymorphisms of TAS1R3 and GNAT3 Genes Are Associated with Patients with Taste Disorder (미각장애와 TAS1R3 및 GNAT3 유전자의 다형성과의 연관성)

  • Bae, Jae-Woong;Kim, Un-Kyung;Kwon, Tae-Jun;Choi, Su-Jin;Ye, Mi-Kyung
    • Journal of Life Science
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    • v.21 no.3
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    • pp.412-416
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    • 2011
  • Taste sensation plays a crucial role in selecting and ingesting foods with different qualities which convey information about their nutrient content and/or safety. Sweetness is one of the five modalities in humans and serves as an energy resource for metabolism. There are reports on allelic polymorphisms which influence perception of sweetness in mice and humans. Since the influence of genetic factors on taste disorder has not been studied, we investigated the association of genetic polymorphisms in TAS1R3 and guanine nucleotide binding protein, alpha transducing 3 (GNAT3) genes and taste disorder. A total of 150 individuals composed of 50 patients with taste disorder and 100 healthy controls were recruited for the study and PCR-mediated directing sequencing method was used to genotype for two different single nucleotide polymorphisms (SNPs) - rs307355 (T>C) and rs35744813 (T>C) in the TAS1R3 gene, and rs7792845 (T>C) and rs1524600 (C>T) in the the GNAT3 gene. The allele and genotype frequencies of rs307355 and rs35744813 in the TAS1R3 gene showed a significant association between patients with taste disorder (p=0.022 and p=0.013 in both of SNPs, respectively). In addition, the frequency of T-T haplotype in the TAS1R3 gene was higher in taste disorder cases than in the controls (OR, 1.93: 95%. CI, 1.09-3.39, p=0.022). In the GNAT3, the genotype frequency of rs7792845 in the patients was also different from the controls (p=0.048), but allele frequency was not significantly associated in either group. Our result provides the frequencies of SNPs and haplotypes of the TAS1R3 and GNAT3 genes for the fundamental information of nutrigenetics in perception of the taste of sweetness in the Korean population. Also, the study suggests that the allelic polymorphisms of TAS1R3 and GNAT3 genes may be useful as a molecular marker for evaluating patients with taste disorder. Further studies with large samples are required to clarify our observation.

The Significance of Ku70 Expression in Patients with Nasopharyngeal Carcinoma (비인강암에서 방사선치료의 예측인자로써 Ku70 발현의 의의)

  • Song Si Yeol;Lee Sang-wook;Yu Eunsil;Cho Kyung-Ja;Park Jin-hong;Kim Sang Yoon;Nam Soon Yuhl;Lee Bong-Jae;Kim Sung Bae;Choi Seung-Ho;Ahn Seung Do;Shin Seong Soo;Choi Eun Kyung
    • Radiation Oncology Journal
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    • v.23 no.1
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    • pp.1-8
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    • 2005
  • Purpose : The objective of this study was to determine whether the expressions of the two components of DNA-dependent protein kinase, Ku70 and DNA-PKcs, influence the response to radiotherapy (RT) and outcome of treatment of non-disseminated nasopharyngeal carcinoma (NPC) in patients who received definitive RT. Materials and Methods : Sixty-six patients with NPC who were treated with radiotherapy alone or with concurrent chemotherapy between June 1995 and December 2001 were divided into groups based on the levels of immunoreactivity for Ku70 and DNA-PKcs in pretreatment biopsy specimens. The over-expression of Ku70 or DNA-PKcs groups Included patients whose biopsy specimens showed at least 50% immunopositive tumor cells; patients in which less than 50% of the tumor cells in the biopsy tissues were immunopositive were placed in the low Ku70 and DNA-PKcs groups. The immunoreactivities for Ku70 and DNA-PKcs were retrospectively compared with the sensitivity of the tumor to radiation and the patterns of therapy failure. Univariate analyses were peformed to determine the prognostic factors that influenced locoregional control of NPC. Results : The five-year locoregional control rate was significantly higher in the low Ku70 group (Ku(-)) (85%) than in the high Ku70 group (Ku(+)) (42%) (p=0.0042). However, there were no differences in the metastases-free survival rates between the two groups (Ku70 (+), 82%; Ku70 (-), 78%; p=0.8672). Univariate analysis indicated that the over-expression of Ku70 surpassed other well-known predictive clinocopathologic parameters as an Independent prognostic factor for locoregionai control. Eighteen of 22 patients who had locoregional recurrences of the tumor displayed an over-expression of Ku70. No significant association was found between the level of DNA-PKcs expression and the clinical outcome. Conclusion : Our data suggest that the level of Ku70 expression can be used as a molecular marker to predict the response to RT and the locoregional control after RT and concurrent chemotherapy in patients with non-disseminated NPC.

Immunohistochemical Detection of p53, erbB-2 and CEA Oncoprotein in Lung Cancer; Clinical Correlations (폐암 환자에서 면역조직화학 염색을 통한 p53, erbB-2, CEA 종양단백 발현과 임상적 의의)

  • Jeong, Seong-Su;Kang, Dong-Won;Lee, Gyu-Seung;Ko, Dong-Seok;Suh, Jae-Chul;Kim, Geun-Hwa;Shin, Kyoung-Sang;Kim, Ju-Ock;Song, Gyu-Sang;Kim, Sun-Young
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.766-775
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    • 1998
  • Background : The prognosis of patients with lung cancer is still poor. Lung cancer exhibits a variable clinical outcome, even in those patients with same stage. Numerous reports suggest that oncogene expression might playa role in explaining the variability of response and survival But many of these reports are still under debate. So we studied the clinical relevance of oncogene expression in Korean lung cancer patients. Immunohistochemistry of p53, erbB-2, CEA expression was performed. Method: From March, 1992 until March, 1997, 120 patients with lung cancer were reviewed. p53, erbB-2, and CEA expression were detected on paraffin-embedded tumor blocks with the use of monoclonal antibodies. The survival and response has correlated with the expressibility of p53, erbB-2, and CEA oncoprotein Results: Overall, the expression rates of p53, erbB-2, and CEA were 33.7%, 59.3%, and 32.6% respectively. Expression rates were not correlated to cell type or stage. Compared with response to chemotherapy, no correlation was found. The expression of p53, erbB-2, or CEA was not correlated with 2-year survival. With simultaneous applications of p53, erbB-2, and CEA, patients with 2 or more expressions also did not show poor response to chemotherapy. Conclusion: We conclude the p53, erbB-2, and CEA expression are clinically less useful in predicting response to chemotherapy or survival.

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