B형 간염바이러스 항원 preS 및 S부위가 포함된 동물세포 발현벡터의 제작

정성균,윤미숙,정대균 경희대학교식량자원개발연구소 2007 硏究論文集 Vol.26 No.1

현재까지 혈장유래 백신이나 재 조합 DNA 기술을 이용한 백신이 HBV 감염을 우수하게 억제하였음에도 불구하고, 성 관련 질병이나 마약중독 환자의 경우 낮은 면역력을 나타내며, HLA haplotype 유전자의 이상으로 인하여 T 세포 면역 반응이 활성화 되지 않는 환자들에게는 제한적인 면역능이 확인되고 있다. 이와 같은 문제점을 해결하기 위하여 기존에 사용하던 S 항원에 Pre S1이나 Pre S2 항원을 첨가하는 새로운 백신 개발에 많은 연구가 진행되고 있다. Pre S2 항원은 HBV 비리온의 표면에 존재하는 표면 단백질이며(Heermann 등, 1984) human serum albumin과 결합 한 후(Krone 등, 1990) 간세포의 표면에 존재하는 human serum albumin receptor를 통하여 HBV를 간세포에 결합시키는 역할을 한다(Pontisso 등, 1989). 또한 pre S2 항원은 현재 S 항원 a 결정자의 HBV의 subtype 별 아미노산 차이에 의한 B 세포 관련 면역성 차이를 극복할 수 있어 heterologous strain의 감염에 따른 비 면역성의 문제를 극복 할 수 있다. 이와 같은 pre S2 항원의 특징을 근거로 하여 본 연구에서는 pre S2와 S 항원을 백신으로 사용하고자 발현벡터인 pIEspHBVS2-dhfr을 제작하였다. 일반적으로 포유동물 세포주에서 고 농도의 단백질을 발현시키기 위한 방법으로 2가지 구별된 벡터를 사용한다. 그 중 하나는 cytotoxic drug인 methotrexate(MTX)와 반응하여 유전자가 증폭되는 dihydrofolate reductase 유전자(dhfr)을 포함하는 발현 벡터를 사용하는 것이며, 다른 하나는 chromatin structure을 조절하는 요소인 matrix attachment regions(MARs)와 scaffold attachment regions(SAR)을 사용하여 높은 transcriptional 활성을 나타내는 유전적 환경을 제공함으로써 유전자 발현을 증가시키는 벡터를 사용하는 것이다(Raju 등, 2001). 또한 포유동물세포에서 외래 유전자 발현의 최대화는 프로모터의 세기와 연관이 있음으로, 우수한 프로모터를 사용하는 것이 가장 필수적이다. 이런 이유로 본 연구에서는 HBsAg유전자 증폭을 위하여 dihydrofolate reductase 유전자(dhfr)와 다양한 포유동물세포에서 강력한 프로모터로 알려져 있는 HCMV immediate-early enhancer promotor를 사용하였다(Kim 등, 2002). 따라서 본 연구에서 제작된 발현벡터인 pIEspHBVS2-dhfr은 dhfr 유전자가 발현시키고자 하는 HBsAg 유전자와 동일한 벡터 내에 존재함으로 이 벡터를 포유동물세포주인 CHO 세포에 형질 전환 시 동시에 CHO 세포 내로 도입되고 또한 염색체 DNA상의 동일 위치에 삽입되게 되어 동시 발현함에 따라 dhfr 선별배지에서 선별된 dhfr 양성 세포주들 모두는 고농도의 HBsAg 단백질을 발현할 것으로 사료된다. A new mammalian cell expression vector for HBsAg, preS and S, was constructed using HCMV imediate early enhancer promoter. In order to develop vaccines against HBV, 10 different plasmids expressing various regions of hepatitis B surface antigen (HBsAg) were constructed. In order to amplify the HBsAg expression, we inserted dihydrofolate reductase (dhfr) gene into the plasmid containing nucleotide 2878 to 846 of HBV genome (pIEsp2878-dhfr).

치과진료실에서의 감염방지 대책에 관한 조사

윤미숙,홍연표,정연강 중앙대학교 의과대학 간호학과 간호과학연구소 1997 중앙간호논문집 Vol.1 No.2

To investigate on infection control in some dental clinics in korea. This study was performed on 300 hygienists who are registered on the association of korea dental hygienist and work at dental clinics from November to December, 1995. Each subjects completed the questionnaire which compased of place of employment of dental hygienist, location of clinics, number of outpatients, educational experience about infection control of dental hygienist on infection control and age of dentists as independent varianbles and use of sterilization methods and protection devices as dependent variables. Data Was expressed as percentage and anlayzed as chi-square method using SPSS/PC + 4.0. There was a significant higher performance rate in use of sterilization methods and protection devices in dental clinics of general hospital than private and public dental clinics. There was no difference in use of sterilization methods and protection devices according to location, number of outpatients of dental clinics and age of dentists. In case of dental hygienists who getting educational experience about infection control, the rate of use of sterilization methods and protective devices was higher than no educational experience. This results suggested that the education about infection control and the follow-up study to investigate he causes of low performance in use sterilizsation methods and protective devices in dental clinics was needed.

DMPO 와 ㆍOH 의 Spin添加生成物에 대한 EPR 分析 및 ㆍOH 의 毒性作用

김정렬,이미녕,박윤창 성균관대학교 기초과학연구소 1989 論文集 Vol.40 No.2

The hydroxyl radical (·OH) generated from the Iron-catalyzed Haber-Weiss reaction was detected. The ability of Fe(Ⅱ) to this reaction depends upon the particular ligands around the metal ion. The formation of hydroxyl radical, spin trapped with 5, 5-dimethyl-1-pyrroline-N-oxide (DMPO), increased with the amount of ATP added in the Fe (Ⅱ)-H_2O_2 system. Specially, when the concentration of H_2O_2 was high, the perhydroxyl radicals were formed from the system. Human erythrocyte ghosts were covalently labeled with 4-maleimido-2,2,6,6-tetramethylpiperidine-N-oxyl(4-Maleimido-Tempo) and their electron paramagnetic resonance(EPR) spectra revealed two major binding environments representing strongly (h_1) and weakly (h_2) immobilized) species. The ratio of h_2/h_1, determined from the respective peak amplitudes, decreased from 14.8 to 5.5 when the labeled ghosts were exposured with hydroxyl radicals produced in the system, whereas lipid peroxidation of erythrocyte ghosts was increased with the concentration of hydroxyl radicals. These effects suggested that the fluidity of membrane and/or protein mobility were decreased as a result of lipid peroxidation and the structural and the functional alterations of membrane were. initiated by hydroxyl radicals.

휴대폰 케이스 가공업체에서 발생한 은피증 1예

탁기홍,윤석환,이미영,신동훈,정인성 대한산업의학회 2006 대한직업환경의학회지 Vol.18 No.4

배경: 은피증은 은의 피부 침착에 의해서 생기는 질환으로, 만성적인 노출에 의해 생기며, 치료법이 없는 영구적인 질병이다. 은의 흡입에 의한 은피증의 보고는 아주 드물며, 우리나라에는 현재까지 관련 보고가 없었다. 이에 휴대폰 도장제의 은 흡입에 의한 은피증을 보고하고자 한다. 증례: 30세 남자근로자로 5년 전부터 휴대폰 부품조립 및 도장업체에서 EMI부서와 가스켓부서에서 전자기파 차단제를 스프레이 도장하는 작업을 하였고, 3년 전부터 서서히 발생한 얼굴과 몸의 회색 및 회청색 피부색 변화를 주소로 내원하였다. 전자기파 차단제는 은, 구리, 용제들로 구성되어 있었다. 피부 조직 검사에서 에크린 한선 기저판(basal lamina)에 미세하고 둥근 흑갈색 과립들이 관찰되어 은피증의 소견을 보였으며, 작업중 노출된 은 흡입에 의한 직업성 은피증으로 진단하였다. 킬레이트 치료는 도움 되지 않으며, 노출 중단 및 자외선 차단 등의 보존적인 방법을 사용하고 있다. 고찰: 은피증은 은의 복용, 흡입, 접촉에 의해 발생한다. 본 증례에서는 은피증을 원인으로 작업시의 은 노출 이외의 은 노출을 발견할 수 없었다. 그러므로, 직업성 은피증으로 사료되며, 은의 첨가제에 의해 은피증이 발생 할 수 있음을 보여준다. 현재 전자기파 차단의 용도로 은과 은 화합물의 사용이 계속되고 있으므로, 이에 대한 대책이 필요하다고 본다. 적절한 치료법이 없기 때문에, 예 방이 가장 중요하며, 보호구 착용, 작업환경 개선, 산업 안전보건교육이 필요하다. Background: Skin contact with, and the breathing in of air containing silver compounds can occur in the workplace. Occupational exposure to silver and its compounds is mainly via airborne dust, metal fumes, and mists of solutions containing silver compounds. Argyria results from increased serum silver levels and the deposition of silver-containing particles in the dermis and mucous membranes. The author reports a patient with generalized argyria, who worked on the manufacture of cellular phone cases. Case report: The patient was a 30 year-old male, with extensive blue-gray discoloration of the whole body, especially sun-exposed area, of 3 years duration. He had been exposed to silver in the processing of EMI for the past 4 years and gaskets for 1 year. EMI is the process by which the internal electromagnetic wave in cellular phone cases are shielded, and gaskets play an important role in shielding the electric clashes between electronic circuits, as well as increasing the durability of cellular phone cases to impact. EMI had been processed manually but this was changed to automatic spraying by robots in 2003. On physical examination, the patient had diffuse blue-gray discoloration of the skin, which was most prominent in the sun-exposed areas of his face, neck and V of the chest. Histopathological examination of a punch biopsy specimen from the posterior neck revealed fine, minute, round, and brown-black granules deposited in the basement membrane zone surrounding the eccrine glands. The laboratory findings were positive for hepatitis B antigen, with a serum silver concentration of 150.3 ppb. From this evidence, he was diagnosed with argyria due to silver exposure. Discussion: The author reports a case of occupational systemic argyria due to inhalation exposure of silver compounds.

비만세포주에서 Histamine 유리에 관한 연구

김형룡,국윤아,신민,이홍수,원진숙,김형민,이영미,정헌택,이혜숙,안녕형 원광대학교 생명공학연구소 1995 생명공학연구소보 Vol.3 No.1

일부 지역 농촌 노인의 철분 영양상태에 관한 연구

신소영,최미경,이윤신,승정자 동아시아식생활학회 1996 동아시아식생활학회지 Vol.6 No.2

The purpose of this study was to examine nutritional status of iron in 27 rural elderly subjects. The daily intake, urinary excretion and serum level of iron were determined by 24-hr food collection method, and 24-hr urine and serum collection for 3 days. The results were summarized as follows. 1. The mean age, height, weight, and blood pressure were 67.7 years, 163.2㎝, 55.9㎏, and 121.1/80.0㎜Hg in men and 69.9 years, 146.5㎝, 47.6㎏, and 108.9/71.1㎜Hg in women, respectively. 2. The daily intake of energy, protein, vitamin A, vitamin B₁, vitamin B₂, niacin, vitamin C, and iron were lower than Korean recommended dietary allowances. 3. The daily iron intake was 7.82㎎. The proportion of heme iron and nonheme iron to the total iron intake were 8.05% and 91.94%, respectively. Major iron sources of the rural elderly were cooked rice, Ko Chu Jang, beef blood, anchovy, soybean paste, and leafy radish. 4. The hemoglobin, hematocrit, serum iron, and total iron binding capacity were 14.3g/dl, 42.9%, 79.4 ㎍/dl, and 268.4 ㎍/dl in men and 12.9g/dl, 38.8%, 92.4 ㎍/dl, and 277.9 ㎍/dl in women, respectively. 5. There was a significant positive correlation between dietary intake and urinary excretion of iron.

標識放鳥에 依한 韓國産 철새 集團의 季節的 分布와 그의 生態(Ⅰ)

元炳旿,禹漢貞,咸奎晃,田美子,尹茂夫 慶熙大學校 1967 論文集 Vol.5 No.-

實驗材料로 토기, 마우스 및 가물치를 使用하여서 致死至近量의 物理的 環境要因 (溫度, 重力, 電擊, 電離放射線 및 非電離放射線), 化學的 環境要因(無機이온) 및 生物學的 環境要因(媒質의 渗透壓)等에 依한 處理가 血液蛋白質의 電氣泳動像을 비롯하여 赤血球, 白血球, 헤마토크리트比, 血色素量, 赤血球脆弱性, 肝臟器官의 組織像 그리고 肝臟 및 腎臟器官重量等에 미치는 效果를 硏究調査하였다. 前記 致死至近量의 環境要因의 處理로 一般的으로 蛋白質代謝, 血液像, 肝臟器官의 組織像 및 肝臟器官重量等에 顯著한 變化를 招來함을 알 수 있었다. 總血淸蛋白質量의 顯著한 變化 및 알부민―굴로부린比의 變化等은 血液內의 渗透效果의 變化를 招來함을 示唆하여 준다. 血淸電氣泳動像의 變化는 前記 環境要因에 依한 處理로 因하여 蛋白質代謝의 異常 卽 肝臟機能의 異常을 招來함을 알 수 있다. 以上의 結果로 미루어 보아 致死至近量이 內的 및 外的의 物理的, 化學的및 生物學的 環境要因에 依한 處理로 因하여 血液蛋白質, 血液像 및 肝臟器官 等의 異常은 生體에 큰 損傷을 가져오며 아울러 生體의 生理的平衡에 有意性的인 變化를 招來한다고 思料된다. Seasonal distribution and ecology of migrant bird populations were studied by mist-netting and banding primarily in the ares of Kyunggido, Korea during 1963-1966. 1. From 6 June to 25 June 1963, 99 birds of 3 species and 3 July 1964 to 31 December 1966, a total of 123, 242 birds of 124 species were banded in Korea. 196 (144 Returns) recoveries of 22 species have been reported in Korea (outside of banding sites) and 7 recoveries of 5 species from abroad. 2. At Taenung, N.E. Seoul, Korea, 11623 birds of the Pied Wagtail and 9,013 birds of the House Swallow were banded in pear orchards. a. Both Wagtail and Swallow are summer resident Wagtail arrives Korea in the beginning of March and Swallows in early April and these gregarious species gather at night into large flocks to roost while juveniles maintain their post-breeding roosts in pear orchard from June to October until the time of Autumn migration. b. Both Wagtaill and Swallow maintain their roosts in the same site but they exhibit different roosting behavior. c. The time at which the Wagtails and Swallows arrive and leave the roost in relation to sunset varies with length of day, weather, however, light intensity mat remain the same. d. The Wagtail gathers around the feeding ground more than 20 kilometers from the roosting site. e. Some Wagtails and Swallows banded the previous year returned and roosted in the orchard. The fact that they repeatedly roost in the same areas suggest that after breeding some birds remain in the Orchard and some of them migrate southward. 3. During July 1964-October 1966, 78, 170 birds of 12 Emberiza species were banded primarily in Kyunggi-do. Emberiza rutila, Emberiza spodocephala, Emberiza tristrami and Emberiza aureola ornata are the dominant fall and spring migrants. In fall they prefer soy-bean, corn and especially millet fields while in spring they are mostly seen on wheat and barley fields. Emberiza rustica is the most abundant species during late fall and winter and they prefer open fields with bushes. Emberiza rutila migrates through Korea in May and from the beginning of August until the and of October. The sex ratio is 100 females to 155 males (11674 ♀, 17761 ♂). More males were caught than females each month except in September when more females were caught. Emberiza spodocephala migrates through Korea from the middle of April through the middle of May and from the meddle of September through October. Emberiza tristrami migrates southward through Korea the first half of May and during October. The sex ratio is 100 female to 140 males(392 ♀, 551 ♂). Emberiza aureola oruata megrates through Korea during May nd from early August until the end of October. Emberiza rustica migrates southward but some of them winter in Korea. The wintering period is from the early October until the end of April. The sex ratio is 100 females to 191 mules (13,450 ♀, 25,687 ♂). Emberiza yessoensis continentals migrates to Korea around the middle of October. Some of them wander in Korea while some of them migrate southward. The wandering period is from the middle of October to the meddle of February. Emberiza cioidos is a permanent resident and breeds in great number in Korea. However, they migrate southward in large flocks during winter and northward during spring. The sex ratio is 100 females to 159 males (497 ♀, 792 ♂). Emberiza e. elegans is a resident and breeds in fairly small number in Korea. but they migrate southward in large flocks around the end of October and northward around April. The sex ratio is 100 females to 280 males(462 ♀, 962 ♂). Emberiza f. fucata is a summer resident, arrives to Korea the middle of April and most of them migrate southward in September. Emberiza chrysophtys migrates southward through Korea in May and from September to October. Emberiza pusilla is a not uncommon transient during spring and autumn and some of it wanders during winter in Korea. Emberiza leucocephala leucocephala Known as a straggler based on the sex specimens but a rare winter visitor. It migrates southward through Korea and some of it wanders around the end of January to the middle of March. 4. Noteworthy records of 21 species observed by the authors and some new species in Korea are given, each with notes on banding and collection records-See text for each species. Observations were made on the feeding habits of nestlings of ten species, Lanius tigrinus, Butorides striatus amurensis, Fhnberiza f. fucata, Motazilla alba leucopsis, Oriolus chinensis diffusns. Accipiter nisus nisosimilis, Mierascelis amaurotis henssent. Garrulus glandarins brandtii, Accipiter solecnsis and Cyanopica eyamts Koreensis. The investigation was made in Kwangnung experimental forest, Kyunggido and the nearby open field by using collar method. Accipiter nisus nisosimiles, Microscelis amaurotis hensoni, Garrulus glandarius brandtii and Cyanopiea cyanus Koreenses are permanent residents and the other six species are common summer residents. The following is the food that these nestlings consumed. 1. Lanius tigrinus The food they consumed was animal matter Composes of; insect larvae-41.5%, insect adults-49.4%, spiders-7.69%, frogs-1.53%. The cicada, Gampsocleis ussuricnsis made up 33.8% of the insect larvae. The cricket, Platyplenra kacmpferi made up 35.4% of the adult insects. Only one species of spiders, Clubiona jucunda was seen, 7.69% of the food. 2. Butorides striatus amurensis The food was animal matters composed of; small freshwater fishes-18.57%, Ranidae-45.71%, others-5.71%. Twenty percent of the small freshwater fishes was Zacco platypus while Hemibarbus logirostris made up 14.28%. Among Amphibians, Rana n. nigromaculata included 22.8%. 3. Emberiza f. fucata The food was amimal matter; insect larvae-63.3%, adult insects-25.64%, others-12.78%, Since 48.1% of the food items was larvae of Pieris rapae, it is most useful for agriculture. 4. Motacilla alba larcopsis The food during their nestling period was animal matter compsed of; insect larvae-30.5%, adult insects-55.4%, spiders-13.9%. The commonest insect larvae were Odonata indet.-22.2%. The commonest adult insects were Syrphidae indet.-16.6%. Of the spiders Lyecsa sp. made up 12.1% and Lycosa astrigera-2.8%. 5. Oriolus chinensis diffusus The food was animal matter composed of; insect larvae-62.36%, adult insects-20.17%, others017.42%. It should be noted that from the initial time of its feeding until leaving it consumed Dendrolimus spectabilis, a noxious forest insect which made up 45.08% of the diet. Thus it is very useful birds eliminating noxious forest insects. 6. Accipiter nisus nisosimilis The food was animal matter composed of; small passerine birds-87.5%, Ranidac-12.05%. It consumed such forest birds as Paridae. 7. Microscelis amauretis hensani The food was primarily insect adults-79.65%, Mollusca-12.15%, vegetable matter-4.05%, insect larvae-2.70%. Araneina-1.35%. Homoptera was 43.35% of adult insects. 8. Garrulus glaudarins brandtii The food during its nestling pcriod was solely animal matter; insect larvae-38.80%, adult insects-28.90%, Araneina-24.87%, adult Amphibia-15.49%. Right afrer hatehing and until leaving its nest, Dendrolimus spectabilis-35.08%, Arancina-24.87%, Clubiona jueunda-6.43% were fed. Therefor 73.48% of the total food was noxious forestry insects. 9. Accipiter soloensis The food during the whole feeding period was primarily Rana n. nigroma-culata 89.09%, but small quantity of Platypleura Kaempferi 8.26%, was also fed. 10. Cyanopica cyanus koreensis The food was composed of; insect larvae-6.11%, adult insects-60.55%, Hyla arborea japonica 12.22%, vegetable matter-1.11%. Gampsocleis ussuriensis-23.39%, Platypleura knempferi-15.55%, and Hyla arborea japoniea-12.22% were the prefered food supplied during the whole feeding period.

Southern Blot Hybridization법을 이용한 악성 림프종의 면역유전형에 관한 연구

전호종,기근홍,김윤신,이미숙,이미자,장원재,장숙진,박영진,정춘해,정종훈,양성훈,이광민 조선대학교 1994 The Medical Journal of Chosun University Vol.19 No.2

Immunogenotyping using gene rearrangement analysis has emerged as a precise laboratory aid in the diagnosis and classification of malignant Iymphoid neoplasms. The lineage and clonality of the malignant Iymphoid neoplasms can be identified by the demonstration of rearrangements of antigen receptor genes of the immunoglobulin and T-cell receptor genes. The analysis of the gene rearrangements on the malignant Iymphoid neoplasms are also useful as a sensitive unique clonal markers to detect early recurrence in patients with malignant Iymphoid neoplasms after treatment. To analyze the sensitivity and specificity of gene rearrangements in the diagnosis of malignant Iymphoid neoplasms. 24 cases of malignat Iymphoma were examined by Southern blot hybridization using CTβ-T cell receptor β chain gene-DNA probe and JH-immunoglobulin heavy chain gene-DNA probe. The results of the immunogenotypings using Southern blot hybridization disclosed high correlation between the immunophenotyping using immunohistochemical stain with monoclonal antibodies (B-cell Iymphoma 84.2%. T-cell Iymphoma 75% ). The analysis of the gene rearrangement of the angioimmunoblastic lymphadenopathy(AILD) and unclassifiable Iymphoma using immunohistochemical stain could resolve the monoclonality and lineage . Rearranged bands to the CTβDNA probe were observed in one case out of 2 cases of AILD. One case of unclassifiable Iymphoma showed rearranged bands to the CTβ DNA probe. There were no rearrangements in reactive follicular Iymphoid hyperplasia and paracortical Iymphoid hyperplasia. In conclusion, DNA gene rearrangement study should be applied to differentiate the clonality and cell lineage in the malignant Iymphoma with indistinctive immunophenotype.

임상 증상이 심하지 않은 심낭 삼출 환자의 장기 추적 결과

배장호,김기식,강미숙,남명희,김미정,김민수,김기영,님창욱,김윤년,김권배,정시전,김여희 대한순환기학회 1999 Korean Circulation Journal Vol.29 No.7

보건간호활동으로 추진한 노인건강증진 프로그램

유인영(In-Young Yoo),조정민(Chung0Min Cho),최영숙(Young-Sook Choi),정문희(Moon-Hee Jung),김미혜(Mi-Hye Kim),유광수(Kwang-Soo You),윤숙례(Suk-Rye Yoon),한명화(Myung-Hwa Han),최선하(Sun-Ha Choi),하영(Young Ha),김규성(Kyu-Sung Kim),이현주 한국보건간호학회 1997 韓國保健看護學會誌 Vol.11 No.1