• Asian Pacific Journal of Cancer Prevention
• /
• 제15권10호
• /
• pp.4211-4215
• /
• 2014

Background: Although roles of genetic polymorphisms of leptin receptor (LEPR) gene in several cancers have been documented, the association between polymorphisms of LEPR and clear cell renal cell carcinoma (CC-RCC) remains unknown. The aim of this study was to explore any relation. Materials and Methods: The study population consisted of 77 patients with CC-RCC and 161 healthy control subjects. Polymorphism analyses of Lys109Arg and Gln223Arg were performed by direct DNA sequencing and PCR-restriction fragment length polymorphism approaches respectively. Results: Comparisons of allelic and genotypic frequencies in Lys109Arg and Gln223Arg showed no significant difference between the cases and controls. However, when evaluating the combined genotype of Lys109Arg and Gln223Arg, risk with GG/GG was increased (OR=1.85, 95%CI=1.04-3.30) and with GA/GG or GG/GA was decreased (OR=0.07, 95%CI=0.01-0.54; OR and 95%CI of the latter could not be calculated for a value of zero). Furthermore, the G-G haplotype frequency of Lys109Arg and Gln223Arg in the cases was higher (OR=1.68; 95%CI=1.02-2.76). In contrast, the A-G and G-A haplotype frequencies in the cases were lower than those in the controls (OR=0.06; 95%CI=0.01 to 0.47; OR and 95%CI of the latter could not be calculated for a value of zero). In addition, the Lys109Arg A allele was in LD with the Gln223Arg A allele (d'=0.9399) in the CC-RCC subjects, but not in the controls. Conclusions: Our data suggest that the GG/GG combined genotype and G-G haplotype of Lys109Arg and Gln223Arg can act as evaluating factors for CC-RCC risk.

• IMMUNE NETWORK
• /
• 제3권3호
• /
• pp.242-247
• /
• 2003

Background: Psoriasis is an inflammatory skin disorder that is characterized by a marked proliferation of keratinocytes, vascular dilation and leukocyte infiltration. Cytokines play important roles in the pathogenesis of inflammatory disorders. An overexpression of proinflammatory cytokines was characterized in psoriasis plaque. Among these cytokines, IL-$1{\beta}$ is major pro-inflammatory cytokine synthesized during the infection and inflammatory process. The IL-1 receptor antagonist (IL-1Ra) competes for the same IL-1 receptor for $IL-1{\alpha}$ and $-1{\beta}$, which prevents activation of the target cells. Three single nucleotide polymorphisms (SNPs) in the IL-$1{\beta}$ gene have been reported at position -31, -511 and +3954. Within the IL-1Ra gene (IL-1RN), there is a variable number of tandem repeats (VNTR) of an 86 bp length in intron 2. These polymorphisms related to cytokine production and associated with various diseases. Methods: We investigated the polymorphisms of IL-1B (promoter -511 and +3954) and IL-1RN on 114 psoriasis patients and 311 healthy normal controls in Korean. We performed PCR-RFLP on single nucleotide polymorphisms (SNPs) of IL-1B (promoter -511 and +3954) and fragment analysis on IL-1RN 86 bp VNTR polymorphism. Results: The frequency of IL-1B $-511^*1$ allele (patients vs. controls; 50.0% vs. 42.3%, RR=1.4) was significantly increased and IL-1B $-511^*2$ allele (patients vs. controls; 50.0% vs. 57.7%, RR=0.7) decreased in psoriasis patients compared to normal controls. We also analyzed the IL-1B -511 polymorphism according to patients' characters (age of onset, sex and family history). The IL-1B -511 alleles were significantly associated in patients with male and family history than health normal controls. There were no significant associations of IL-1B +3954 and IL-1RN polymorphisms with psoriasis patients. Conclusion: These results suggest that the polymorphism of IL-1B -511 could be genetic susceptibility to psoriasis in Koreans.

• IMMUNE NETWORK
• /
• 제3권3호
• /
• pp.219-226
• /
• 2003

Background: Phage display is the most widely used technique among display methods to produce monoclonal antibody fragment with a specific binding activity. Having a large library for efficient antibody display/selection is quite laborious process to have more than $10^9$ members of transformants. To overcome these limitations, several in vitro selection approaches have been reported. Ribosome display that links phenotypes, proteins, directly to genotype, mRNA, is one of the in vitro display methods. Ribosome display can reach the size of scFv library up to $10^{14}$ molecules and it can be further diversified during PCR steps. To select the high affinity scFv from one pot library, we established ribosome display technique by modifying the previously reported eukaryotic translation system. Methods: To establish the antibody selection system by ribosome display, we used 3D8, anti-DNA antibody. A 3D8 scFv was synthesized in vitro by an in vitro transcription-translation system. The translated 3D8 scFv and the encoding 3D8 mRNA are connected to the ribosome. These scFv-ribosome-mRNA complexes were selected by binding to their specific antigens. The eluted mRNAs from the complexes are reverse transcribed and re-amplified by PCR. To apply this system, antibody library from immunized mouse with terminal protein (TP)-peptide of hepatitis B virus DNA polymerase TP domain was also used. This TP-peptide encompasses the 57~80 amino acid residues of TP. These mRNA/ribosome/scFv complexes by our system were panned three times against TP-peptide. The enrichment of antibody from library was determined by radioimmunoassay. Results: We specifically selected 3D8, anti-DNA antibody, against ssDNA as a model system. The selected 3D8 RNAs sequences from translation complexes were recovered by RT-PCR. By applying this model system, we enriched TP-peptide-specific scFv pools through three cycles of panning from immunized library. Conclusion: We show that our translating ribosome complexes are well maintained and we can enrich the TP-specific scFv pools. This system can be applied to select specific antibody from an antibody library.

• Tuberculosis and Respiratory Diseases
• /
• 제67권1호
• /
• pp.8-13
• /
• 2009

Background: The insulin receptor substrate-1 (IRS-1) is the primary docking molecule for the insulin-like growth factor I receptor (IGF-IR), and is required for activation of the phosphatidylinositol 3'-kinase (PI3K) pathway. IRS-1 activation of the (PI3K) pathway regulates IGF-mediated survival, enhancement of cellular motility and apoptosis. Therefore, we attempted to ascertain whether IRS-1 genetic variations affect an individual's risk for non-small cell lung cancer (NSCLC). Methods: Two-hundred and eighteen subjects, either diagnosed with NSCLC or control subjects, were matched by age, gender and smoking status. Genomic DNA from each subject was amplified by PCR and analyzed according to the restriction fragment length polymorphism (RFLP) profile to detect the IRS-1 G972R polymorphism. Results: The frequencies of each polymorphic variation, in the control population, were as follows: GG=103 (94.5%) and GR=6 (5.5%); for the NSCLC subjects, the genotypic frequencies were as follows: GG=106 (97.2%) and GR=3 (2.8%). We could not demonstrate statistically significant differences in the genotypic distribution between the NSCLC and the control subjects (p=0.499, Fisher's Exact test). The relative risk of NSCLC, associated with the IRS-1 G972R polymorphic variation, was 1.028 (95% CI; 0.63~9.90). In addition, we found no differences between polymorphic variants with regard to the histological subtype of NSCLC. Conclusion: We did not observe any noteworthy differences in the frequency of the IRS-1 G972R polymorphism in NSCLC patients, compared to control subjects. These results suggest suggesting that, in our study population, the IRS-1 G972R polymorphism does may not appear to be associated with an increased risk of NSCLC.

• Journal of Gastric Cancer
• /
• 제8권3호
• /
• pp.113-119
• /
• 2008

목적: TFF1 유전자의 -2bp에 위치하고 있는 단일염기다형성의 유전자형 및 대립형질 빈도와 H. pylori 감염 및 위암발생 민감성과의 연관성을 조사하고자 하였다. 대상 및 방법: 2000년 1월부터 2003년 12월까지 위 샘암종으로 근치적 수술을 시행 받은 167명과 정상 건강인 299명의 DNA를 이용하였다. TFF1 유전자의 단일염기다형성의 유전자형은 중합효소연쇄반응 후 제한효소를 이용한 제한 분절길이다형성과 single strand conformation polymorphism의 방법으로 분석하였다. 또한, H. pylori 감염은 Giemsa 염색으로 조사하였다. 결과: 정상 건강인의 TFF1 단일염기다형성에 대한 유전자형 및 대립형질의 빈도를 위암 환자군과 비교한 결과 통계적으로 의의가 없었다(P=0.715 & P=0.595). 위암 환자군을 성별로 나누어 정상 건강인과 비교한 경우에도 통계적 의의를 발견할 수 없었다. 또한, 위암 조직의 조직학적 소견 및 환자 연령과 TFF1 단일염기다형성의 유전자형 및 대립형질 빈도를 조사한 결과, 위암 조직의 조직학적 소견 및 연령은 TFF1 유전자 단일염기다형성과 무관하였다(P=0.088 & P=0.551). 한편, H. pylori 감염은 모두 39예에서 관찰되었는데 TFF1 유전자형과는 통계적으로 연관성이 없었다(P=0.7200). 결론: TFF1 유전자의 -2bp에 존재하는 단일염기다형성은 TFF1 단백의 합성에 큰 영향이 없으며 H. pylori 감염과 위암의 발생과는 무관하다는 것을 의미한다.

• IMMUNE NETWORK
• /
• 제2권4호
• /
• pp.242-247
• /
• 2002

Background: Tumor necrosis factor-alpha (TNF-$\alpha$) is known to play an important role in various conditions such as inflammation, autoimmunity, apoptosis, insulin resistance and sleep induction. Five single nucleotide polymorphisms (SNPs) have been known to affect the transcriptional activities of TNF-$\alpha$: -1,031T/C, -863C/A, -857C/T, -308G/A and -238G/A. Methods: We have investigated 5 SNPs of the promoter region of TNF-$\alpha$ gene, the distribution of 5-locus TNF-$\alpha$ haplotypes, and their haplotypic associations with previously typed HLA-A, -B and -DRB1 loci in 107 healthy unrelated Koreans. TNF-$\alpha$ SNPs were typed using PCR-single-strand conformation polymorphism (SSCP) and PCR-restriction fragment length polymorphism (RFLP) methods. Results: The allele frequencies of -1,031C, -863A, -857T, -308A, and-238A, which are known as the high-producer-type, were 19.3%, 15.9%, 14.0%, 5.9%, and 2.9%, respectively. The frequency of -308A allele, known to be associated with autoimmune diseases, was 5.9% in Koreans which was lower than Caucasians (14~17%) and somewhat higher than Japanese (1.7%). Five most common TNF-$\alpha$ haplotypes (-1,031/-863/-857/-308/-238) comprised over 95% of total haplotypes: TCCGG (58.4%), CACGG (14.8%), TCTGG (13.7%), TCCAG (5.3%), and CCCGA (3.1%). Strong positive associations (P<0.001) were observed between TCCGG and B62; between CACGG and B51, $DRB1^*0901$; between TCTGG and B35, B54, B59, $DRB1^*1201$; and between TCCAG and A33, B58, $DRB1^*0301$, $DRB1^*1302$. Five most common extended haplotypes (>3%) comprised around 16% of total haplotypes: A33-B58-TCCAG-$DRB1^*1302$, A24-B52-TCCGG-$DRB1^*1502$, A33-B44-TCCGG-$DRB1^*1302$, A24-B7-TCCGG-$DRB1^*0101$, and A11-B62-TCCGG-$DRB1^*0406$. The distribution of extended HLA and TNF-$\alpha$ haplotypes showed that most of HLA haplotypes were almost exclusively associated with particular TNF-$\alpha$ haplotypes. Conclusion: The results obtained in this study would be useful as basic data for anthropologic studies and disease association studies in Koreans.

• 대한족부족관절학회지
• /
• 제17권2호
• /
• pp.130-135
• /
• 2013

Purpose: Preshaped Locking compression plate(LCP) has holes with fixed angle between screw and plate and have advantage firm fixation because it has stability of angular and axial deformity. We evaluated usefulness of LCP after open reduction and internal fixation in distal fibular fracture. Materials and Methods: Between December 2011 and May 2012, 23 patients with fracture of distal fibula were followed up at least 12 months underwent open reduction and internal fixation with LCP. There were 15 males and 8 females with a mean age 39.8(20~69) years. According to Danis-Weber classification, there were 20 cases of type B and 3 cases of type C. There were 13 cases of isolated lateral malleolus fractures, 1 case of bimalleolar fracture, 6 cases of trimalleolar fractures and 3 cases of distal tibia fractures with proximal fibula fracture. Intraoperatively, we assessed whether preshaped LCP fit lateral margin of distal fibula or not and evaluated quality of reduction and postoperative complications. The cases were analyzed by radiological bone union time and clinical results according to the criteria of Meyer Results: Of all cases, complete bone union was achieved and average radiological bone union time was 7.3(6~12) weeks. The clinical results were excellent in 18 cases(78%), good in 5 cases(22%). There were 5 cases of plate with 3 holes, 13 cases of plate with 4 holes, 2 cases of plate with 5 holes, 1 case of plate with 6 holes and 2 cases of plate with 7 holes. The average number of screws at proximal fragement was 2.5 and at distal fragment was 3.5. In 14 cases (60.8 %), we needed re-bending of plate because the distance between plate and lateral cortical margin of distal fibula was more than 5 mm at anteroposterior X-ray after reduction. All cases have anatomical reduction and there were no complications of wound infections. There were no complaint about hardware irritation. Conclusion: At fractures of distal fibula,preshaped LCP had a excellent stability although far cortex was not fixed with screw and bending of plate. And there are less complications of hardware irritation and wound problems. But, Some complement would be needed because there were no complete fitting between precontour of LCP and lateral cortical margin of distal fibula.

• 한국양식학회지
• /
• 제8권3호
• /
• pp.209-220
• /
• 1995

미꾸라지의 간으로부터 핵을 분리하여, 저농도 염추출 및 제한효소 처리로 핵기질(nuclear matrix)을 분리하였다. 분리된 핵기질을 Proteinase K로 분해한 후, phenol-chloroform 추출로 크기가 약 0.3kb-15kb의 분포를 나타내는 핵기질 부착 DNA (nuclear matrix attachment regions : MARs)를 얻었다. 효모 URA 3 유전자를 가진 2.13 kb Eco47 III 단편을 제한효소 Ssp I 으로 절단된 pUC19 플라즈미드 벡타에 결합시켜, ARS (autonomously replication sequence) 클로닝을 위한 pURY19 플라즈미드 벡타를 만들었다. 이 pURY19 벡타는 Saccharomyces cerevisiae내에서 독립적으로 복제할 수 없기 때문에, 물고기의 효율적인 발현 벡타 개발을 위해, 이 system을 이용하여, S. cerevisiae내에서 독립적으로 복제 가능한 미꾸라지의 ARS를 클로닝하고자 하였다. 분리 된 MARs를 pURY19 벡타에 결합시 킨 다음, E. coli $DH5\alpha$에 형질전환시켜 $pURY19N_{l-62}$를 얻었다. MAR Libraries $(pURY19N_{1-62})$를 각각 $Ura^-\;S.\;cerevisiae$에 형질전환시켜, S. cerevisiae내에서 독립적으로 복제 가능한 M. mizolepis 유래의 복제원점들 (ARSs)을 분리하여, Sanger's dideoxy-chain termination method로 염기서열을 분석하였다. 염기서열 분석결과 모든 clones들은 AT-rich하였으며, 특히 $pURY19N_6$에는 ARS concensus sequence, Topoisomerase II consensus, near A-box, 그리고 T-box들이 존재하였다.

• 한국양식학회지
• /
• 제9권4호
• /
• pp.445-452
• /
• 1996

어류 및 폐류내에 내재하는 내재성 ${\beta}-galactosidase$의 활성도를 분석함으로서 외래유전자의 이식시 기초자료로 활용하고자 어류 6종 및 패류 4종을 대상으로 본 실험을 행하였다. 어류에 있어서의 X-gal 염색 결과 혈청 및 근육을 제외한 모든 조직에서 모두 positive ( + ) 염색 양상을 보였다. 패류에 있어서도 어류와 같은 양상을 보여 근육을 제외한 모든 조직에서 모두 positive 염색 양상을 보였으며 더욱이 참굴에 있어서는 폐각근 및 근육에 있어 매우 진한 염색 반응을 보였다. 근육으로 부터 DNA를 추출한 후 PCR을 수행한 결과 모든 종에서 positive (+) 결과를 나타내었으며, 어패류간 그리고 각 개체간 차이는 없었다. 각 장기별 ${\beta}-galactosidase$의 활성도 측정결과 모든 종에 있어서 혈청의 활성도는 무시할 만한 수준이었고, 근육에서 가장 낮게 나타났다. 미꾸라지와 잉어에서는 신장에서 가장 높은 활성을 보였고, 틸라피아는 소화관에서 가장 높은 값을 보였다. 해산어인 넙치의 경우 돌가지미, 문치가자미의 경우에 비해 간에서 활성이 높았다. 그러나 돌가자미 및 문치가자미인 경우 신장에서 가장 높은 값을 보였다. 패류에 있어서는 소화맹낭에서 가장 높은 활성값을 나타내었고 폐각근 및 외투막에서 매우 낮은 활성값을 나타내었다. 그러나, 참굴에 있어서는 참전복, 피조개 및 진주조개 보다는 상대적으로 폐각근 및 외투근에서 높은 활성을 나타내었다.

• 대한소아치과학회지
• /
• 제43권2호
• /
• pp.200-206
• /
• 2016

상아질, 백악질, 치수를 포함하는 파절로 정의되는 치근파절은 영구치에서 발생하는 외상 중 0.5-7%를 차지하며, 대부분 11-20세군의 상악 전치부에 호발한다. 영구치 치근파절의 처치는 기본적으로 변위된 치관부 파절편을 정복시키고 고정하는 것이다. 60-80%는 치수생활력이 유지되나 치수괴사나 염증성 치근흡수의 징후가 나타난다면 근관치료를 시행하게 되며 대부분에서 치근부 파절편의 치수생활력은 유지되기 때문에 근관치료는 치관부 파절편에 한하여 수행한다. 그러나 치관부 파절편에서 적절한 apical stop을 얻는 것은 어렵다. 의도적 재식술이란 통제된 환경에서 의도적으로 치아를 발거한 후 구강 외에서 치근단 치료를 시행하고 재식립하는 방법으로 완벽한 근관치료와 수복을 목표로 한다. 통상적인 근관치료가 실패한 경우, 기존의 수복물이 존재하거나 석회화된 근관으로 인해 재근관치료가 어려운 경우, 공간적으로 접근이 불가능하여 치근단 수술을 시행하지 못하는 증례에서 의도적 재식술이 계획될 수 있다. 본 증례에서는 이전의 외상으로 인해 석회화된 근관을 보이는 상악 중절치에서 발생한 수평 치근파절을 치료하기 위해 의도적 재식술을 이용하였고 임상적, 기능적으로 만족할만한 결과를 얻었기에 이를 보고하는 바이다.