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The expression and cellular localization of phospholipase D isozymes in the developing mouse testis
민도식,Taekyun Shin,김승준,Heechul Kim,Yongduk Lee,Jin Won Hyun,이영호,Min Kyoung Shin 대한수의학회 2007 Journal of Veterinary Science Vol.8 No.3
To examine the involvement of phospholipase D (PLD)isozymes in postnatal testis development, the expression of PLD1 and PLD2 was examined in the mouse testis at postnatal weeks 1, 2, 4, and 8 using Western blot analysis and immunohistochemistry. The expression of both PLD1 and PLD2 increased gradually with development from postnatal week 1 to 8. Immunohistochemically, PLD immunoreactivity was detected in some germ cells in the testis and interstitial Leydig cells at postnatal week 1. PLD was mainly detected in the spermatocytes and residual bodies of spermatids in the testis after 8 weeks after birth. The intense immunostaining of PLD in Leydig cells remained unchanged by postnatal week 8. These findings suggest that PLD isozymes are involved in the spermatogenesis of the mouse testis. To examine the involvement of phospholipase D (PLD)isozymes in postnatal testis development, the expression of PLD1 and PLD2 was examined in the mouse testis at postnatal weeks 1, 2, 4, and 8 using Western blot analysis and immunohistochemistry. The expression of both PLD1 and PLD2 increased gradually with development from postnatal week 1 to 8. Immunohistochemically, PLD immunoreactivity was detected in some germ cells in the testis and interstitial Leydig cells at postnatal week 1. PLD was mainly detected in the spermatocytes and residual bodies of spermatids in the testis after 8 weeks after birth. The intense immunostaining of PLD in Leydig cells remained unchanged by postnatal week 8. These findings suggest that PLD isozymes are involved in the spermatogenesis of the mouse testis.
새로이 개발된 비탁법을 이용한 혈청 저밀도 지단백 콜레스테롤의 직접 측정법의 평가
민도식,김덕언,박일규 한양대학교 의과대학 1999 한양의대 학술지 Vol.19 No.1
The association between elevated levels of low-density lipoprotein cholesterol (LDL-C) and increased risk of premature coronary heart disease has been clearly demonstrated. Currently, most of laboratories rely on indirect measurement of LDL-C by using Friedewald equation which are not applied in samples of non-fasting or over 400mg/dL of triglyceride level. For these reasons, a newly developed direct LDL-C assay by turbidimetric method was evaluated and compared with Friedewald equation. Also, we evaluated the quality control materials which are made in our laboratory. Total cholesterol, triglyceride, HDL-C, direct LDL-C and Friedewald LDL-C are measured on commercial control sera, two patient's pooled sera and patient sera using automated chemistry analyzer (Hitachi-747, Japan). Direct LDL-C assay using LDL Cholesterol homogeneous (Boeringer Mannheim, Germany) was performed using automated chemistry analyzer (Hitachi-747, Japan). Within-run precisions of Friedewald LDL-C and direct LDL-C were 113.83±3.70 mg/dL (3.31%) and 124.83±2.25 mg/dL (1.80%), respectively in control sera. Between-run precision were 112.53±2.60 (2.31%) and 120.64±2.39 (1.98%) repectively. The correlation coefficient between Friedewald LDL-C and direct LDL-C methods in 64 patient's samples was 0.87 and the linear regression equation was Y = 0.593X + 40.868 at triglyceride levels below 400 mg/dL. There was no significant decrease in LDL-C determined by direct LDL-C assay for non-fasting versus fasting sera and their values were 97.2±42.9 mg/dL, 95.9±41.0 mg/dL respectively. In two pooled sera, intra-assay CV of lyophilized serum which was restored by 1 mL distilled water was 1.84% and the other serum which was thawed after freezing was 17.90%. Precisions of direct LDL-C using LDL Cholesterol homgeneous were better than calculated LDL-C values using Friedewald equation in control sera. There was a good correlation between two methods with triglyceride levels below 400 mg/dL. Thus this direct LDL-C assay is recommended in place of Friedewald equation which has a few limitation for use. And the lyophiized sera from pooled sera is recommended to use in quality control instead of company-made control sera which are expensive and have many shortcoming.
류마티스 관절염 환자에서 Shared Epitope의 임상적 의의
민도식 ( Doh Sik Minn ),김신규 ( Think You Kim ) 대한류마티스학회 2001 대한류마티스학회지 Vol.8 No.1
Objective: The application of DNA sequencing and molecular-based typing to detect HLA-DRB1 alleles showed that those associated with rheumatoid arthritis (RA) shared a consensus amino acid sequence (QKRAA or QRRAA) at positions 70-74 of the third hypervariable region of the HLA-DRB1 chain. This is defined as the so-called `shared epitope`. Many studies reported that shared epitope was associated with RA susceptibility and disease severity. Also, DRB1*0401 and DRB1*0404 alleles confer genetic predisposition to RA in Caucasian. We studied the frequency of shared epitope, DRB1*04, DRB1*0401, and DRB1*0404 in Korean RA patients using monoclonal antibodies. We also tried to investigate the influence of these factors on susceptibility and severity in RA patients and to evaluate the method. Methods: RA patients were 32 persons with classical or definite RA who attended the Hospital for Rheumatic Diseases, Hanyang University Hospital, Seoul, Korea. We separated lymphocytes from whole blood and used indirect immunofluorescence staining method using four monoclonal antibodies (Terra Nova, Canada). Results: The frequency of DRB1*04 and shared epitope were 56% and 56%, respectively. That of DRB1*0401 and DRB1*0404 were positive in 19% and 9% of all patients, respectively. There were no association between share epitope and disease severity. Conclusion: The shared epitope is expressed with high frequency, as many as DR4 frequency in Korean RA patients. But the frequency of DRB1*0401 and DRB1*0404 is low different from Caucasian. The used method is simple and easy to screen shared epitope.
Jurkat T 면역세포에서 Phosphoinositides 의 가수분해를 증가시키는 약용식물 추출물의 검색
민도식(Do Sik Min),이영한(Young Han Lee),백석환(Suk Hwan Baek),서판길(Pann Ghill Suh),류성호(Sung Ho Ryu) 한국응용약물학회 1996 Biomolecules & Therapeutics(구 응용약물학회지) Vol.4 No.2
Activation of the T lymphocytes results in a variety of early biochemical events ultimately leading to cell proliferation and lymphokine production. Stimulation of the signal transduction cascade in T cells through the T cell receptor coincides with activation of the phosphatidylinositol-phospholipase C (PI-PLC) pathway. Therefore, we have established a model system to screen immuno-simulator that can increase the hydrolysis of phosphoinositides in human T cell leukemia Jurkat cells. As a result of screening from herbal medicine extract, 4 extracts (Olibanum, Ephedrae Herba, Real Gar, Saussureae Radix) were found to increase the production of inositol phosphates. All the active fraction from the four kinds of extract were eluted in a different retention time on C-18 HPLC and these active fraction also showed difference in cell specificity. And all the active fractions increased DNA synthesis in T cell. Therefore, it is suggested that the active fraction among 4 extracts might contain a compound having different properties one another.