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인슐린 농도 변화에 따른 인슐린 내재화 ( Internalization ) 와 Degradation 의 변화
박원근(Won Kun Park),최웅환(Woong Hwan Choi),한인권(In Kwon Han),김선우(Sun Woo Kim),정운원(Woon Won Jung),문인걸(In Gul Moon) 대한내과학회 1988 대한내과학회지 Vol.34 No.4
N/A This study was conducted to investigate the effect of ambient concentration of insulin on insulin internalization and degradation in human erythrocytes. The absolute amount of internalized insulin was quantitated by multiplying the concentration of insulin by the percent specifically bound and internalized (by acid extraction method, PBS, pH 5.7) at the various insulin concentration (1.4, 2.5, 12.4, 112.4 ng/m1). The insulin degradation products (IDP) was assayed by using Sephadex G-50 column, HPLC with radioisotope detector in human erythrocytes which were incubated (37℃) at the various insulin concentration (1.43, 2.76, 14.76, 26.60ng/ml). Insulin internalization (ng/ml) increased in human erythrocytes with increasing concentration of insulin (3hr incubation time, 0.336 at 1.4, 0.275, at 2,5, 3.844 at 12.4, and 24.728ng/ml at 112.4ng/ml, 4hr incubation time, 0. 476, at 1.4, 0.925 at 2.5, 4.712 at 12.4 and 38.216ng/ml at 112.4ng/ml). Insulin degradation products (%) increased (2hr incubation time, 20.9 at 1,43; 38.0 at 2.76 and 38.0% at 14.76ng/ml) but no more than at a level of insulin concentration (2.76ng/ml) in normal human erythrocytes. In diabetic human erythrocytes, insulin degradation products (%) also increased (3br incubation time, 14.1 at 1.43, 19.0 at 2,76, 23.0 at 14.76, and 23.2% at 26.60ng/ml, 4hr incubation time, 39.4 at 1.43, 46.9 at ~2.76, 52. S at 14.76 and 53.0% at 26.60ng/ml) but no more than at a level of insulin concentration (14.76ng/ml), This result indicated that the ambient concentration of insulin may play an important role in the regulation of intracellular processing of insulin.
Type 2 DM 환자에서 적혈구를 이용한 인슐린 내재화율과 Degradation Products 에 관한 연구
최웅환(Woong Hwan Choi),박원근(Won Kun Park),한인권(In Kwon Han),김선우(Sun Woo Kim),정운원(Woon Won Jung),문인걸(In Gul Moon) 대한내과학회 1988 대한내과학회지 Vol.34 No.3
N/A We studied insulin internalization and degradation at 37℃ in human erythrocytes from patients with type II DM and normal subjects. The internalization of 125I-Insulin in human erythrocytes was studied by using an acid extraction technique (pH 5.7, PBS) from 54 patients with type II DM and 14 normal subjects. Insulin degradation products was assayed with using Sephadex G-50 column, HPLC with radioisotope detector from 4 patients with type II DM and 14 normal subjects. The maximal rate of insulin internalization (Incubation time: 4hours, at 37℃) was decreased in patients with type II DM [51.40±14.67% (±SD) VS 80.23±7.73 % (±SD). P<0.001]. The maximal degradation products of insulin (Incubation time: 4hours, at 37℃) was decreased in patients with type II DM [62.50±10.50% (±SD) VS 87.75±6.66% (SD). p<0.001]. In conclusion, insulin internalization and degradation in human eryhrocytes from patients with type II DM are significantly reduced. These defect may be related to the cellular insulin resistance present in these patients.
정운원 청주대학교 보건의료과학연구소 2014 보건의료과학연구 Vol.3 No.1
Over the past decades, the development and application of molecular diagnostics has been made in all fields of laboratory medicine as in vitro diagnostics. Although conventional methods are mainly used, there is an increasing trend towards molecular diagnostics. These techniques are superior to conventional ones in rapid detection, higher sensitivity and specificity. Polymerase chain reaction(PCR)-based systems has dominated in various nucleic acid-based techniques, such as PCR, sequencing, DNA microarray, lab-on-a-chip(LOC) and etc. PCR-based systems can detect the etiologic agents through the amplification of disease-related gene directly from clinical samples and have been undergone various modification for more rapid, accurate and applicable to diagnostic tests. Sequencing analysis ensures exact identification and better characterization of the etiologic agents. Some like array-based systems offer the multiparameter results where many pathogen-related markers and gene mutations can be analysed simultaneously. The development of LOC chip should allow point-of-care testing for the revolutionary improvement of healthcare. Along with the development of proteomic or metabolomic-based tests, More advanced diagnostic tools will expand the applications of molecular diagnostic testing. The user-friendly automation about both the high throughput and the use of minimal quantity of sample makes these technologies more widely available.
정운원 청주대학교 보건의료과학연구소 2015 보건의료과학연구 Vol.4 No.1
In molecular diagnostics, the methodological development and application of detecting and quantifying nucleic acid has been made over the past decades. Nucleic acid analysis based polymerase chain reaction(PCR) has been dominated among various techniques, such as PCR, sequencing, microarray and others. Detection and quantification of disease-related genes gives us a important information that may be used to predict disease progression, signs of infection and the efficacy of therapeutic agent. traditional PCR is based on exponential amplification and quantification of nucleic acids may be determined by comparing the number of amplification cycles and amount of PCR product to those of a reference specimen. However, many factors can complicate the calculation of quantification, resulting in uncertainties and inaccuracies. Digital PCR is considered to have overcome the critical problems of insufficient quantification in traditional PCR. The difference between dPCR and traditional PCR lies on the measuring method for nucleic acids amounts. Absolute quantification through clonal amplification is a key factor in digital PCR. It has many possible applications which include the detection and quantification of low-level pathogens, viral load, rare allele, copy number variation, and relative gene expression. Along with the more advanced development in technology of digital PCR, the clinical applications of digital PCR will be greatly expanded. Therefore, the present review describes the principle, technological types and clinical applications of digital PCR as useful molecular diagnostic tool.
DNA Microarray를 이용한 인유두종 바이러스(Human Papilloma Virus:HPV)의 진단
정운원,이승관,이창규,조경진,김성욱 高麗大學校 倂設 保健大學 保健科學硏究所 2001 保健科學論集 Vol.27 No.1
Human papillomavirus(HPV) has been known as one of the important pathogenic agent in uterine cervical carcinoma. The molecular works such as PCR enable the detection of large number of HPV genotypes obtained from viginal swab. Many of the PCR-based methods for HPV detection involve an amplification step followed by any of a number of methods for distinguishing different HPV types. In this study, we adopted the DNA chip technology enabling a HPV type-specific differentiation both low-risk group(type-6, 11, 34, 40, 42, 43, and 44) and high-risk group(type-16, 18, 31, 33, 35, 39, 45, 51,52,54, 56, and 58). MY09/MY11 and GP5+/GP6+primers covered LI region are used in nested PCR to improve PCR amplification. HPV type-specific probes for DNA chip were modified with NH2-C6, followed by spotting on silylated slides, washing slides and hybridization with each PCR products. Of 163 DNA samples chosen randomly, 42 samples were negative, 8 ones for low-risk group of HPV and 96 ones for high-risk group of HPV. Especially, co-infections with various HPVs were shown in 17 samples. A recent study found that multiple HPV is a factor in persistent HPV infection, resulting in the development of cervical dysplasia. This result emphasized the necessity to detect multiple HPV infection. The application of DNA-chip to determinate specific HPV typing will be a stronger candidate than any other PCR-based methods. Furthermore, the sequencing data of the positive PCR products were shown no discrepancy with DNA chip results. This means that DNA chip is very useful tool for both HPV detection and typing.