大氣汚染物質의 變異原性에 關한 硏究 : 마우스 母體 및 胎仔의 小核試驗을 中心으로

韓東宣,車喆煥,張聖薰,裵思相 고려대학교 의과대학 1989 고려대 의대 잡지 Vol.26 No.3

This Study aimed at identifying the cytogenic effect of air pollutants in pregnant mouse and her fetus. Samples of air pollutants were collected by the high volume air sampler in several sites of Seoul from May through December in 1988. Total suspended particulates (TSP) were measured and those of benzene and methanol were extracted and dissolved in dimethyl sulfoxied (DMSO). Those were injected into peritoneal cavity by the concentration and the micronucleated polychromatic erythrocyte (MNPCE) was observed in the bone marrow of mother mice. IN addition, the liver of fetus was extracted and MNPCE was observed. Transplacental genetic toxicity due to air pollutants in both cases was measured in the end. 1. In case of the tar concentration equivalent to those residing in the volume of 100m³ of air, the MNPCEs in the mother's bone marrow were 0.57±0.15% and 0.70±.20% in Shinchon; 0.27±0.17% and 0.40±0.13 In Jungrung and in the underground shopping center ; 0.33±0.10% and 0.55±0.13% in the subway station. The MNPCEs measured in the sample of the tunnels, of which tar concentration was equivalent to that in 50m³ and 100m³ of air, were 0.86±0.24% and 1.06±0.17% in tunnel A and 0.70±0.15% and 0.96±0.23% in tunnel B; those were the highest MNPCE in the samples of mother mice. 2. The MNPCEs measured in the liver of featus were 0.60±0.25% and 0.80±0.15% in Shinchon. In Jungrung and the underground shopping center, the MNPCEs in the liver of fetus were the same as those in mothers. In the sample of the subway station, the MNPCEs in the liver of fetus, 0.73±0.20%, were higher than in mothers only in the 200? tar concentration. The MNPCEs in the 50m³ and 100m³ tar concentration were 0.87±0.10% and 1.27±0.27% in tunnel A, and 0.68±0.41% and 1.07±0.23% in tunnel B. It showed that the MNPCEs only in the 100m³ tar concentration were higher than those in mothers.

이황화탄소 폭로 근로자의 요중 N-Acetyl-β-Glucosaminidase Activity 및 Total protein에 관한 연구

趙浚吉,李恩一,車喆煥,張聖薰 고려대학교 의과대학 1992 고려대 의대 잡지 Vol.29 No.1

The results of renal function studies among 45 workers exposed to carbon disulfide as measured bv total protein in a 24-hour urine and N-Acetyl-β-Glucosaminidase (NAG) activity in both 24-hour and spot urine, were further analyzed by fundoscopic examination for evidence of retinopathy. By Measuring spot urine NAG activity in 69 healthy workers, a comparative analysis of spot urine NAG activity was made between healthy workers and those exposed to carbon disulfide and the results were as follows: 1. Of the workers exposed to carbon disulfide, 53.3% had retinopathy, 53.3% showed increased NAG activity above reference value, and 20.0% had proteinuria. These rates tended to increase as the age of the workers and their duration of exposure increased. 2. For workers exposed to carbon disulfide and having retinopathy, the mean value of NAG activity and total protein amount in 24-hour urine were significantly higher (p<0.05), as compared to those having normal retina. 3. Mean values of spot urine NAG activity in healthy workers, those exposed to carbon disulfide with normal retina, and those having retinopathy were 199.1μM of 4-MU/mgCr., 499.1μM of 4-MU/mgCr., and 902.7μM of 4-MU/mgCr., respectively, showing a statistically significant difference.

요중 Protein 및 N-Acetyl-β-Glucosaminidase의 생물학적 변동에 관한 연구

朴園,李恩一,車喆煥,張聖薰 고려대학교 의과대학 1992 고려대 의대 잡지 Vol.29 No.1

The purpose of this study was to evaluate the biological variations of N-acetyl-β-glucosaminidase activity, beta-2 microglobulin, alpha-1 microglobulin, total protein, and microalbumin in spot urine specimens, all of which have been used as a screening test for the evaluation of renal dysfunction. The study was conducted from June to August of 1991, with analysis-evaluation of the 24-hour urine specimens with that of 3 spot-urine specimens and first morning specimen. A total of 14 healthy college student volunteers (non-exposed group) and 42 workers exposed to carbon disulfide (exposed group)were selected as the study subjects. The results were as follows ; 1. The mean values of N-acetyl-β-glucosaminidase activity and alpha-1 microglobulin in 24 hour urine specimens in the exposed group were higher than those in the non-exposed group. 2. Coefficient of variation among spot urine specimens of N-acetyl-β-glucosaminidase in exposed group was the lowest among the results of 5 different types of urine tests. 3. The correlation coefficients between the values in 24 hour urine specimens and those in spot urine specimens were above 0.9 for total protein. microalbumin, and alpha-1 microglobulin. The correlation coefficients between the NAG activities in 24 hour urine specimens and those in spot urine specimens were relatively high. 4. Of the total 25 workers who showed a higher N-acetyl-β-glucosaminidase activity than the reference data for the 24 hour urine specimens, the number of workers excessive reference data for the spot urine specimens was 23(92%), and 22(88.9%) for the first morning urine specimens.

Benz(a)anthracene이 마우스태자 간조직 세포의 소핵출현에 미치는 경태반효과

成耆天,張聖薰,車喆煥 고려대학교 의과대학 1989 고려대 의대 잡지 Vol.26 No.1

In order to adduce the cytogenic effect of Benz(a)anthracene(B(a)A) in pregnant mice through placental barrier to fetus, 125㎎/㎏, 250㎎/㎏, 500㎎/㎏, of B(a)A were administrated to groups of mice 16th pregnancy day About 20 hrs later, freauency of micronuclei in polychromatophilic normoblasts of liver tissues extracted from fetus was measured At the same time, 200 ㎎/㎏ and 400 ㎎/㎏ of Benzo(a) pyrene(B(a)P) were administered with same procedure to compare the frequencies between in cases of B(a)A and B(a)P administration. Especially incase of B(a)A administration, frequencies of their appearances in bone marrows of mother mice and livers of fetuses were compared. On the other hand, pathological changes of fetus liver caused by B(a)A and B(a)P in mothers were observed through microscopic examination in order to reconfirm and compare their transplacental effects. As the results, cytogenic toxicity for feture caused of B(a)A was confirmed. It means that some amount of B(a)A which was administered to pregnant mice could be transmitted to fetus through placental barriers Mutagenicity caused of B(a)A was higher in fetus than mother mouse. The cytogenic toxicity caused of B(a)A both for mother mouse and fetus was lower than that of B(a)p.

모 형광등업체에 있어서의 공기중 수은농도 및 요중 수은량 조사

박승회,김광종,장성훈,차철환 大韓産業醫學會 1989 대한직업환경의학회지 Vol.1 No.2

In order of interpret the relationship among many variables, urinary mercury and the same time mercury concentration in air were measured for 254 workers of a fluorescent lamp manufacturing factory. And monthly consumption amounts, numbers of monthly producing fluorescent lamps, numbers of inferior lamps numbers of breakage lamps, numbers of vacuum pumps and frequencies of mercury infusion were also investigated from January 1988 to March in 1989. The results were as follows; 1. On the mercury concentration in air by producing line and by working site, all of them were exceed the threshold limit value (0.05mg/㎥). 2. The highest mercury concentration in air by sampling was found at the floor of workplace (0.334mg/㎥) and next were at vacuum exhaustion prmp (0.183mg/㎥), and breathing zone of workers(0.103mg/㎥) in order. 3. The highest mean of the mercury concentration in urine by producing line was the automatic exhausting line (80.8㎍/1) and next were high speed exhausting line (72.4㎍/1), and Manual exhausting line (35.8㎍/1) in order. Distribution of workers of the mercury concentration in urine more than 200㎍/1 by working site was the highest (10%) in the vacuum exhaustion pump part and next were sealing part (6.6%), packing part (4.6%), and stem part (4.1%) in order. 4. The correlation coefficient between mercury concentration in urine and in air was the highest(0.302) and next were numbers of breakage lamps (0.223), frequencies of mercury infusion (0.223), numbers of inferior lamps (0.205) in order.

당뇨병 환자에서 혈장 Thrombin-Antithrombin Ⅲ 및 Plasmin-α_2-Plasmin Inhibitor 복합체의 임상적 의의

김경욱,김은숙,정상수,윤수지,박우일,이준희,남수연,안철우,문병수,김경래,차봉수,송영득,임승길,이현철,허갑범 대한당뇨병학회 2002 Diabetes and Metabolism Journal Vol.25 No.5

연구배경:당뇨병 환자에서 혈액응고 및 섬유소용해 체계의 이상경향이 있어 그 결과로 여러 혈관합병증의 발생위험이 높다는 사실은 널리 알려져 있다. 그 기전은 아직 확실히 밝혀지지 않았으나, 고혈당으로 인한 혈장 단백질들의 비효소성 당화작용이나 산화성 스트레스로 인한 유리 라티칼 작용으로 응고항진이나 섬유소용해 활성의 저하를 유발하는 것으로 생각되고 있다. 최근 응고 및 용해인자와 그 억제자의 복합체들의 증가가 이 상태를 비교적 예민하게 반영한다고 알려져 있다. 방법:본 연구에서는 당뇨병 환자 101명과 정상 대조군 20명에서 혈장내 thrombin­antithrombin complex(TAT)와 plasmin­α₂­plasmin inhibitor complex(PIC)를 측정하여 비교하고, 당뇨병 환자에서 미세혈관 합병증과 대혈관합병증의 유무에 따른 차이와, 이미 혈관 질환의 위험인자로 알려져 있는 인자들간의 상관성을 알아보고자 하였다. 결과:1. 환자의 분포를 살펴보면 혈관합병증이 있는 군은 85명, 혈관합병증이 없는 군은 16명이었고, 평균연령은 각각 57.9±14.1세, 49.9±16.6세로 혈관 합병증이 있는 군에서 더 나이가 많았고, 체질량지수는 23.2±3.4㎏/㎡, 24.1±3.4㎏/㎡로 두 군간 유의한 차이는 없었다. 또 두 군간의 혈압 및 HbA1c, 공복혈당 및 인슐린과 C­peptide, 총 콜레스테롤, 중성지방, HDL­콜레스테롤, Lp⒜는 유의한 차이가 없었고, 미세혈관합병증이 있는 군에서 당뇨병의 유병기간이 길었다. 2. TAT 및 PIC의 농도는 정상 대조군에서는 2.8±1.2 ng/mL, 240.4±69.7 ng/mL이었고, 당뇨병 환자군에서는 9.5±22.6 ng/mL, 472.2±258.7 ng/mL이었다. TAT와 PIC 모두 당뇨병 환자군에서 정상 대조군에 비해 유의하게 증가되어 있었고(p<0.001), TAT/PIC ratio는 두 군간 차이가 없었다. 3. 당뇨병 환자의 혈관합병증에 따른 TAT 및 PIC, fibrinogen 농도는 합병증이 없는 군은 각각 4.1±2.4ng/mL, 362.2±272.0ng/mL, 322.7±102.4mg/mL으로 PIC와 fibrinogen의 증가를 보였으나, 연령을 보정한 후에는 통계학적 유의성은 없었다. 또 대혈관 합병증군에서는 각각 6.0±4.9 ng/mL, 507.4±321.6 ng/mL, 427.1±194.7 mg/dL이었으며 미세·대혈관 합병증군에서는 10.4±6.4 ng/mL, 484.8±269.7 ng/mL, 388.4±132.4 mg/dL으로 TAT의 증가를 보였으나 역시 연령을 보정한 후에는 통계학적 유의성은 없었다. 4. 미세혈관합병증군에서 HbA1c(>8%)가 높은 군의 PIC 농도가 유의하게 높았고(p=0.049), 대혈관합병증군에서 HbA1c(>8%)가 높은 군의 총 콜레스테롤 농도가 유의하게 높았다(p=0.042). 5. 총 당뇨병 환자군에서 PIC는 fibrinogen과 HbA1c와 양의 상관관계를, BMI와 음의 상관관계를 보였으며(r=0.47, 0.31,-0.25), 혈관 합병증이 없는 당뇨병 환자군에서만 TAT는 HbA1c와 양의 상관관계를 보였다(r=0.67). 결론:이상의 결과에서 혈장 TAT 및 PIC 농도는 당뇨병 환자에서 정상 대조군에 비해 의미있게 증가되어 있었고, 당뇨병 환자군에서는 연령의 증가와 유병기간이 혈액응고항진 및 용해의 장애에 큰 역할을 함을 알 수 있었으며, 총 당뇨병 환자군에 PIC와 HbA1c와 양의 상관관계를, BMI와 음의 상관관계를 보였으며 혈관 합병증이 없는 당뇨병 환자군에서만 TAT는 HbA1c와 양의 상관관계를 보였다. 따라서 당뇨병 환자에서 혈액응고 및 용해의 장애가 동반되어 있다고 볼 수 있으며, 혈장 TAT 및 PIC는 혈관합병증으로의 진행을 예측하는 지표로서 유용하리라 생각된다. 또 혈당조절정도와 상관성이 있으므로 혈당조절후에 추적검사를 시행하여 합병증의 예방이 가능한지 추후 연구가 필요하리라 생각된다. Background : Abnormality of coagulation and fibrinolystic system is known as a predisposing factor of vascular complication in diabetes. Although the pathogenesis is not well known, non-enzymatic glycation reaction and the increase in production of free radicals due to an increased oxidative stress may be linked to the hypercoagulibility and hypofibrinolytic activity. As indices of abnormality in coagulation and firinolysis in peripheral blood, plasma thrombin-antithrombin Ⅲ complex (TAT) and plasmin-α_2-plasmin inhibitor complex (PIC) were measured. The purpose of this study was to clarify whether hypercoagulability exists in diabetic patients with or without vascular complication. Methods : In our study, we measured plasma thrombin-antithrombin Ⅲ compelx (TAT) and plasmin-α_2-plasmin inhibit or complex (PIC) in 101 diabetic subjects and 20 controls. Comparing TAT and PIC levels in diabetic microvascular complication group, diabetic macrovascular complication group and controls, we examined correlation between risk factors associated with diabetic vascular complication. Results : 1. The group with diabetic vascular complication was older than group without complication. There was no significant difference in BMI, blood pressure, HbA_ic, blood sugar level, insulin, C-peptide, serum creatinine, total cholesterol, triglyceride, HDL-cholesterol, Lp (a) between two groups. The group with diabetic microvascular complication had longer duration of diabetes. 2. Concentration of TAT and PIC were 2.8±1.2 ng/ mL, 240.4±69.7 ng/ mL in controls and 9.5±22.6 ng/ mL, 472.2±258.7 ng/ mL in diabetic patients, respectively. TAT and PIC were significantly higher in diabetic patients than in control (p<0.001). But TAT/PIC ratio was no significant difference between two groups. 3. In diabetic patients, concentration of TAT and PIC and fibrinogen were respectively 4.1±2.4 ng/ mL, 362.2±272.0 ng/ mL, 322.7±102.4 mg/ dL in group without vascular complication and 5.3±4.1 ng/ mL, 529.5±258.7 ng/ mL, 374.9±106.2 mg/ dL in group with microvascular complication, which group had increase in PIC and Fibrinogen but no significance after correction of age. Concentration of TAT and PIC and Fibrinogen were 60.±4.9 ng/ mL, 507.4±321.6 ng/ mL, 427.1±194.7 mg/ dL in macrovascular complication, and 10.4±6.7 mg/ mL, 484.8±269.7 ng/ mL, 388.4±132.4 mg/ dL in combined vascular complication which group showed increase of TAT but also had no significant increase after correction of age. 4. In diabetic microvascular complication patients, group of high HbA_1c (>8%) (p=0.049) had significant high PIC concentration. In diabetic macrovascular complication patients, group of high HbA_1c (>8%) (p=0.042) had significant high total cholesterol concentration. 5. In all diabetic patients, PIC was positively correlated with fibrinogen and HbA_1c and negatively correlated BMI (r=0.47, 0.31, -0.25). Only in daibetic patients without angiopathy, TAT was positively correlated with HbA_1c (r=0.67). Conclusion : In this study, plasma TAT and PIC concentration significantly increased in diabetic patients compared with controls, and PIC was increased in group with microvascular complication, TAT were increased in group with combined micro macrovascular complication. However, there was no significance relationship existed when correctinf for age. PIC was correlated with HbA_1c. TAT was correlated with HbA_1c only in the group without angiopathy. Abnormality of coagulation and fibrinolysis were combined in diabetes, plasma TAT and PIC can be used as an index of vascular complication. Also we found the correlation with the degree of the blood glucose control. Therefore we need follow up study for the possibility of prevention of vascular complication after controlling the blood glucose to age-matched patients (J Kor Diaabetes Asso 25:354~363, 2001).

Expression of the HSV - 1 ( F ) Glycoprotein B Gene in Insect Cells Infected by HcNPV Recombinant

(Soung Chul Cha),(Hyun Kang),(Sook Yeon Lee),(Gap Ju Park),(Hyung Hoan Lee) 한국미생물생명공학회 2000 Journal of microbiology and biotechnology Vol.10 No.3

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Articles / High Level Production of Glycoprotein H of HSV-1 (F) Using HcNPV Vector System

(Hyun Kang),(Soung Chul Cha),(You Jin Han),(In Ho Park),(Min Jung Lee),(Si Myung Byun),(Hyung Hoan Lee) 생화학분자생물학회 2000 BMB Reports Vol.33 No.6

The Herpes simplex virus type 1 (HSV 1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 by and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV 1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

High Level Production of Glycoprotein H of HSV-1 (F) Using HcNPV Vector System

Kang, Hyun,Cha, Soung-Chul,Han, You-Jin,Park, In-Ho,Lee, Min-Jung,Byun, Si-Myung,Lee, Hyung-Hoan Korean Society for Biochemistry and Molecular Biol 2000 Journal of biochemistry and molecular biology Vol.33 No.6

The Herpes simplex virus type 1 (HSV-1) strain F glycoprotein H (gH) gene in the pHLB-4 plasmid was recombinated into a baculovirus expression vector (lacZ-HcNPV) to construct a recombinant virus GH-HcNPV expressing gH. The sequences of gH and its expression were analyzed. The gH gene was located in the 6.41 kb BglII fragment. The open reading frame (ORF) of the gH gene was 2,517 bp and codes 838 amino acid residues. Insect cells infected with this recombinant virus synthesized a high level of the matured and gX-gH fusion protein with approximately 112 kDa. The fusion gH protein was localized on the membrane of the insect cells as seen by using immunofluorescence assay and accumulated in the cultured media by the SDS-PAGE and immunoprecipitation assays. The amino acid sequence presents additional characteristics compatible with the structure of a viral glycoprotein: signal peptide, putative glycosylation sites and a long C-terminal transmembrane sequence. Antibodies raised in mice to this recombinant protein recognized viral gH and neutralized the infectivity of HSV-1 in vitro. These results demonstrate that it is possible to produce a mature protein by gene transfer in eukaryotic cells, and indicate the utility of the HcNPV-insect cell system for producing and characterizing eukaryotic proteins. Furthermore, the neutralizing antibodies would appear to protect mice against HSV; accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.

Poster Presentation Abstracts ; DNA Vaccine of Glycorpotein B Gene of Herpes Simplex Virus Type 1 against HSV

(Min Jung Lee),(Soung Chul Cha),(Hyun Kang),(Hyung Hoan Lee),(Jae Kyung Cho) 한국응용약물학회 1999 학술대회 Vol.1999 No.2