• Title/Summary/Keyword: 분자 생물학적 분석

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Development of Species-Specific PCR to Determine the Animal Raw Material (종 특이 프라이머를 이용한 동물성 식품원료의 진위 판별법 개발)

  • Kim, Kyu-Heon;Lee, Ho-Yeon;Kim, Yong-Sang;Kim, Mi-Ra;Jung, Yoo Kyung;Lee, Jae-Hwang;Chang, Hye-Sook;Park, Yong-Chjun;Kim, Sang Yub;Choi, Jang Duck;Jang, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.347-355
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    • 2014
  • In this study, the detection method was developed using molecular biological technique to distinguish authenticity of animal raw materials. The genes for distinction of species about animals targeted at Cytochrome c oxidase subunit I (COI), Cytochrome b (Cytb), and 16S ribosomal RNA (16S rRNA) genes in mitochondrial DNA. The species-specific primers were designed by that Polymerase Chain Reaction (PCR) product size was around 200 bp for applying to processed products. The target 24 raw materials were 2 species of domestic animals, 6 species of poultry, 2 species of freshwater fishes, 13 species of marine fishes and 1 species of crustaceans. The results of PCR for Rabbit, Fox, Pheasant, Domestic Pigeon, Rufous Turtle Dove, Quail, Tree Sparrow, Barn Swallow, Catfish, Mandarin Fish, Flying Fish, Mallotus villosus, Pacific Herring, Sand Lance, Japanese Anchovy, Small Yellow Croaker, Halibut, Jacopever, Skate Ray, Ray, File Fish, Sea Bass, Sea Urchin, and Lobster raw materials were confirmed 113 bp ~ 218 bp, respectively. Also, non-specific PCR products were not detected in compare species by species-specific primers. The method using primers developed in this study may be applied to distinguish an authenticity of food materials included animal raw materials for various processed products.

Prevalence and Patterns of Congenitally Missing Teeth among Pediatric Patients Aged 8 - 16 in Pusan National University Dental Hospital (부산대학교 치과병원에 내원한 8 - 16세 환자의 선천성 치아 결손 유병률 및 유형 평가)

  • Eunjin Kim;Soyoung Park;Eungyung Lee;Taesung Jeong;Jonghyun Shin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.50 no.2
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    • pp.179-191
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    • 2023
  • The purpose of this study was to investigate the prevalence and patterns of congenitally missing teeth in permanent teeth excluding third molars, in patients aged 8 to 16 years who visited Pusan National University Dental Hospital from January 2010 to February 2021. This retrospective study evaluated tooth agenesis and the pattern of missing teeth represented by the tooth agenesis code by reviewing panoramic radiographs and electronic medical records of 11,759 patients, including 5,548 females and 6,211 males. The prevalence of congenitally missing teeth was 10.74% (females 11.95%, males 9.66%, p < 0.0001). Patients with tooth agenesis had an average of 2.22 missing teeth, and congenitally missing teeth occurred more frequently in the mandible (8.39%) than in the maxilla (4.52%, p < 0.0001). The mandibular second premolar (58.19%) was the most frequently missing tooth. The second premolar was the most frequently missing tooth in all quadrants (30.10%, 31.67%, 43.14%, and 35.59%) when a single tooth was absent, while the first and second premolars were the most commonly absent teeth (11.69%, 11.47%, 5.94%, and 5.24%) when two or more teeth were missing. In the relationship between maxillary-mandibular antagonistic quadrants and full mouth, the 1st to 4th place of the missing patterns were all involved with the 1st and 2nd premolars. This study can be clinically helpful in establishing a treatment plan for patients with missing teeth. In addition, it can be used as basic data for molecular biological research to find out the relationship between tooth agenesis and specific genes.

Correlation of p53 Protein Overexpression, Gene Mutation with Prognosis in Resected Non-Small Cell Lung Cancer(NSCLC) Patients (비소세포폐암에서 p53유전자의 구조적 이상 및 단백질 발현이 예후에 미치는 영향)

  • Lee, Y.H.;Shin, D.H.;Kim, J.H.;Lim, H.Y.;Chung, K.Y.;Yang, W.I.;Kim, S.K.;Chang, J.;Roh, J.K.;Kim, S.K.;Lee, W.Y.;Kim, B.S.;Kim, B.S.
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.4
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    • pp.339-353
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    • 1994
  • Background : The p53 gene codes for a DNA-binding nuclear phosphoprotein that appears to inhibit the progression of cells from the G1 to the S phase of the cell cycle. Mutations of the p53 gene are common in a wide variety of human cancers, including lung cancer. In lung cancers, point mutations of the p53 gene have been found in all histological types including approximately 45% of resected NSCLC and even more frequently in SCLC specimens. Mutant forms of the p53 protein have transforming activity and interfere with the cell-cycle regulatory function of the wild-type protein. The majority of p53 gene mutations produce proteins with altered conformation and prolonged half life; these mutant proteins accumulate in the cell nucleus and can be detected by immunohistochemical staining. But protein overexpression has been reported in the absence of mutation. p53 protein overexpression or gene mutation is reported poor prognostic factor in breast cancer, but in lung cancer, its prognostic significance is controversial. Method : We investigated the p53 abnormalities by nucleotide sequencing, polymerase chain reaction-single strand conformation polymorphism(PCR-SSCP), and immunohistochemical staining. We correlated these results with each other and survival in 75 patients with NSCLC resected with curative intent. Overexpression of the p53 protein was studied immunohistochemically in archival paraffin- embedded tumor samples using the D07(Novocastra, U.K.) antibody. Overexpression of p53 protein was defined by the nuclear staining of greater than 25% immunopositive cells in tumors. Detection of p53 gene mutation was done by PCR-SSCP and nucleotide sequencing from the exon 5-9 of p53 gene. Result: 1) Of the 75 patients, 36%(27/75) showed p53 overexpression by immunohistochemical stain. There was no survival difference between positive and negative p53 immunostaining(overall median survival of 26 months, disease free median survival of 13 months in both groups). 2) By PCR-SSCP, 27.6%(16/58) of the patients showed mobility shift. There was no significant difference in survival according to mobility shift(overall median survival of 27 in patients without mobility shift vs 20 months in patients with mobility shift, disease free median survival of 8 months vs 10 months respectively). 3) Nucleotide sequence was analysed from 29 patients, and 34.5%(10/29) had mutant p53 sequence. Patients with the presence of gene mutations showed tendency to shortened survival compared with the patients with no mutation(overall median survival of 22 vs 27 months, disease free median survival of 10 vs 20 months), but there was no statistical significance. 4) The sensitivity and specificity of immunostain based on PCR-SSCP was 67.0%, 74.0%, and that of the PCR-SSCP based on the nucleotide sequencing was 91.8%, 96.2% respectively. The concordance rate between the immunostain and PCR-SSCP was 62.5%, and the rate between the PCR-SSCP and nucleotide sequencing was 95.3%. Conclusion : In terms of detection of p53 gene mutation, PCR-SSCP was superior to immunostaining. p53 gene abnormalities either overexpression or mutation were not a significant prognostic factor in NSCLC patients resected with curative intent. However, patients with the mutated p53 gene showed the trends of early relapse.

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A Study of Microsatellite Instability in Primary Small Cell Lung Cancers by Microsatellite Analysis (원발성 소세포폐암에서 Microsatellite 분석을 이용한 Microsatellite 불안정화에 대한 연구)

  • Cho, Eun-Song;Chang, Joon;Park, Jae-Min;Shin, Dong-Hwan;Kim, Se-Hoon;Kim, Young-Sam;Chang, Yoon-Soo;Cho, Chul-Ho;Kwak, Seung-Min;Lee, Jun-Gu;Chung, Kyung-Young;Kim, Sung-Kyu;Lee, Won-Young;Kim, Se-Kyu
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.2
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    • pp.180-190
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    • 2000
  • Background: Genomic instability, which is manifested by the replication error(RER) phenotype, has been proposed for the promotion of genetic alterations necessary for carcinogenesis. Merlo et al. reported frequent microsatellite instability in primary small cell lung cancers. However, Kim et al. found that instability occurred in only 1% of the loci tested and did not resemble the replication error-positive phenotype. The significance of microsatellite instability in the tumorigenesis of small cell lung cancer as well as the relationship between microsatellite instability and its clinical prognosis was investigated in our study. Methods: Fifteen primary small cell lung cancers were chosen for this study. The DNAs extracted from paraffin-embedded tissue blocks with primary tumor and corresponding control tissue were investigated. Forty microsatellite markers on chromosome 1p, 2p, 3p, 5q, 6p, 6q, 9p, 9q, 13q, and 17p were used in the microsatellite analysis. Results: Thirteen(86.7%) of 15 tumors exhibited LOH in at least one of the tested microsatellite markers. Three of 13 tumors exhibiting LOH lost a larger area in chromosome 9p. LOH was shown in 72.7% on chromosome 2p, 40% on 3p, 50% on 5q, 46.7% on 9p, 69.2% on 13q, and 66.7% on 17p(Table 1). Nine(60%) of 15 tumors exhibited shifted bands in at least one of the tested microsatellite markers. Nine cases exhibiting shifted bands showed altered loci ranging 2.5~52.5%(mean $9.4%\pm16.19$)(Table 2). Shifted bands occurred in 5.7% (34 of 600) of the loci tested(Table 2). Nine cases with shifted bands exhibited LOH ranging between 0~83.3%, and the median survival duration of those cases was 35 weeks. Six cases without shifted bands exhibited LOH ranging between 0~83.3%, and the median survival duration of those cases was 73 weeks. There was no significant difference between median survival durations of the two groups(p=0.4712). Conclusion: Microsatellite instability as well as the inactivation of several tumor suppressor genes may play important roles in the development and progression process of tumors. However, the relationship between microsatellite instability and its clinical prognosis in primary small cell lung cancer could not be established.

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