• Journal of Periodontal and Implant Science
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• 제23권2호
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• pp.300-314
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• 1993

Periodontal disease research has been focused on understanding the immunopathologic mechanisms which may operate in the development and maintenance of peiodontal inflammatory changes. Immunologic and inflammatory responses may relate to the etiology and pathogenesis of periodontal disease. In order to research immunopathology of periodontal disease, previous investigators have spent much time on the distribution of lymphocyte subpopulations and NK cells but they have spent less time on the changes of those cells to the periodontal disease severity. The purpose of study was performed to investigate the changes of the distribution of T lymphocytes, B lymphocytes, T lymphocyte subsets, and Natural Killer cells in the gingival epithelium and connective tissue of the periodontal disease with the various clinical parameters including Gingival Index, Sulcular Bleeding Index, and pocket depth. Gingival tissues were obtained from 25 patients with different severity of periodontal disease. Serial cryostat sections displaying a cross section of gingiva were labelled with monoclonal antibody for pan T cells, T cytotoxic/suppressor cells, T helper/inducer cells, pan B cells, and NK cells were develped using an avidin-biotin-peroxidase system. Lymphocyte populations were enumerated in repeatable fields from gingival section. 1. T cells were more increased at grade 1 and 3 than at grade 0 of gingival index (p<0.05). Helper T cells and NK cells were significantly increased at grade 1, 2, 3 than at grade 0(p<0.05). 2. T cells were more decreased at grade 3 and 4 than at grade 1 of sulcular bleeding index (p<0.01, p<0.05). Especially, Natural Killer cells were significantly increased at grade 1, 2, 3, 4 than at grade 0 (p<0.05, p<0.001). 3. The ratios of helper T/suppressor T cells were more decreased at grade 4 than at grade 0 and at grade 4 than at grade 2 of sulcular bleeding index (p<0.05, p<0.05). 4. Helper T cells were significantly decreased at grade II and III than at grade I, however the Natural Killer cells showed a increasing tendency with the increase of the pocket depth, there were no significant differences between each grade of pocket depth. 5. The ratios of helper T/suppressor T cells were tended to be decreased with the increase of the pocket depth, there were no significant differences between each grades of pocket depth. There was a very weak change in the distribution of T lymphocytes, B lymphocytes, T lymphocyte subsets, and Natural Killer cells in the gingival epithelium and connective tissue of the periodontal lesion with the various clinical parameters including gingial index, sulcular bleeding index, and pocket depth. But, the number of T lymphocytes and Natural Killer cells were significantly changed in gingival index and sulcular bleeding index.

• Parasites, Hosts and Diseases
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• 제31권1호
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• pp.49-56
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• 1993

현재 유구낭미충증을 혈청학적으로 진단하는 데에는 낭액항원을 이용한 특이항체검사법으로 micro-ELISA를 널리 이용하고 있다. 이 실험은 효소부착 이차항체를 사용하는 micro-ELISA방법 대신 민감도가 뛰어난 것으로 알려진 ABC-ELISA나 Protein-ELISA로 바꾸어 사용하면 검사의 민감도를 보다 개선할 수 있는지를 알기 위하여 실시하였다. ABC-ELISA에 의한 항체검사는 낭액항원 단백질 $2.5{\;}\mu\textrm{g}/ml$, 혈청 희석 1:10,000, biotinylated no-hmn IgG 1:100,000 회석, peroxidase conjugated streptavidin 1:40,000 희석 및 2,2-azino-di(3-ethylbenztlllazoline) sulfnnlc acid발색제를 사용하는 조건으로 실시하였고 415 nm에서 흡광도를 측정하였다. Protein A-ELISA는 항원단백질 $2.5{\;}\mu\textrm{g}/ml$, 혈청은 1:200 회석, HRP-Protein A 1:20,000 희석 및 ABTS 발색제를 사용하는 조건으로 실시하고 415 nm에서 흡광도를 측정하였다. 유구낭미충증 환자 115 명의 혈청을 검사한 바 민감도는 micro-ELISA 81.7%, Protein A-ELISA 82.6%, ABC-ELISA 86. 1%이었다. 다른 기생충성 질환자, 비기생충성 질환자 및 대조군 등 165명 혈청에서 특이도는 각각 88.5%, 93.3%, 93.8%이었다 세 가지 ELISA방법에 의한 항체가 사이에는 상관계수 0.84~0.86의 높은 상관관계가 성립하였다. 이상의 결과 ABC-ELISA는 micro-ELISA에 비하여 민감한 혈청학적 방법이고 실제 유구낭미충 특이항체 검사상의 특이도에서도 차이가 있다는 것을 알 수 있었다. 따라서 ABC-ELISA는 유구낭미충증의 혈청학적 진단에서 혈청 및 뇌척수액 등 검체를 적게 사용할 수 있다는 장점이 있다는 것을 알 수 있었다.

• 대한임상검사과학회지
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• 제39권2호
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• pp.128-135
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• 2007

This study was designed to investigate the correlation between the expression rate of p53 and p21 proteins by immunohistochemical staining and tumor prognostic factors including the tumor size, histological differentiation and Dukes' stage of tumor prognostic factors in colon cancer, and to acquire necessary data for the presumption of diagnosis, treatment and prognosis of colon cancer patients. From January 2000 to January 2003 at Hanyang University Guri Hospital, the paraffin blocks of 35 patients diagnosed with colon cancer whose pathologic reports were possible to review were selected. Harris hematoxylin & eosin (H&E) staining and immunohistochemical staining by ABC (Avidin Biotin Conjugate) method were performed. The histological differentiation grade and stage were classified according to the classification of the World Health Organization (WHO) and modified Dukes's stage from H&E staining. The expression rate of p53 and p21 proteins were analyzed by immunohistochemical staining. The results was analyzed statistically by SPSS (Windows version 8.0). As a result, the expression rate of p53 protein was 11.4% (4 cases) in clear differentiation, 48.6% (17 cases) in moderate differentiation, and 17.1% (6 cases) in poor differentiation. In other words, the poorer the differentiation, the higher the expression rate of p53 protein (p<0.05). The expression rate of p21 was 17.1% (6 cases) in clear differentiation, 40.0%(14 cases) in moderate differentiation, and 8.6% (3 cases) in poor differentiation, According to the progression of histological malignant degeneration, the expression rate of p21 protein decreased distinctively (p<0.05). However, the correlation between the two above mentioned proteins and the tumor-size and Dukes' stage was not of statistical significance. In the comparison of the expression rate of p53 protein with that of p21 protein, in 10 cases, p53 protein expression was positive while p21 protein expression was negative, and in 6 cases, p53 protein expression was negative whereas p21 protein expression was positive. Consequently a statistically significant inverse correlation between the expression rate of p53 protein and that of p21 protein was observed (p<0.05). In conclusion, we found a significant correlation between histological differentiation and the expression rate of p53 and p21 proteins (p<0.05), and a significant inverse correlation between the expression rate of p53 protein and that of p21 protein (p<0.05). Also, it could be confirmed that the over expression of p53 and p21 proteins is closely associated with the occurrence of colon cancer and its progress. Therefore, it is thought that this study may be greatly beneficial to the presumption of diagnosis, treatment and prognosis of colon cancer patients.

• 대한수의학회지
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• 제39권3호
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• pp.575-582
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• 1999

Morphometrical analysis of chicken Cryptosporidium baileyi in various stages of life cycle in the bursa of Fabricius were carried out by electron microscope to establish a differential point for identification of C baileyi. By avidin-biotin complex method, protozoans of the bursa of Fabricius were identified Cryptosporidium spp. The size and area on each developmental stages of C baileyi, as measured by Morphomat 10 attached to electron microscope were as follows. Trophozoites' size with range of $3.21{\pm}0.70{\times}2.49{\pm}0.59{\mu}m$, area with range of $118.82{\pm}41.92{\mu}m^2$; meronts' size $3.99{\pm}1.07{\times}2.96{\pm}0.52{\mu}m$, area $210.11{\pm}57.11{\mu}m^2$; merozoites' size $1.98{\pm}0.43{\times}0.60{\pm}0.18{\mu}m$, area $24.10{\pm}5.97{\mu}m^2$; microgametes' size $1.36{\pm}0.83{\times}0.50{\pm}0.23{\mu}m$, area $20.23{\pm}6.73{\mu}m^2$; macrogametes' size $4.57{\pm}0.65{\times}4.02{\pm}0.55{\mu}m$, area $258.37{\pm}51.83{\mu}m^2$; oocytes' size $4.39{\pm}0.56{\times}3.44{\pm}0.50{\mu}m$, area $187.21{\pm}62.68{\mu}m^2$. In conclusion, the size and area on each developmental stages of Cryptosporidium baileyi is different from that of other Gryptosporidia spp. It suggests, with considering tissue tropism and life cycle, morphometrical analysis can be quite a good way to identify C baileyi.

• Restorative Dentistry and Endodontics
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• 제24권2호
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• pp.372-380
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• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide(CGRP) containing nerve fivers after pulp exposure in rats. The Spague-Dawley rats weighing about 250 - 300g were used. The animals were devided into normal control group and experimental groups. Experimental animals were sacrified on 2, 4, 7, 10 days after pulp exposure. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde plus 0.1M phosphate buffer (pH 7.4). Serial frozen $50{\mu}m$ thick sections were cut with a cryostat. In the immunohistochemical staining procedure, the rabbit CGRP antibody was used as a primary antibody. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated anti-rabbit IgG as a secondary antibody and incubated in ABC (avidin-biotin complex), The sections were visualized by 0.05% 3.3 diaminobenzidine tetrahydrochloride. The results of this study were as follows: 1. In control group, CGRP containing nerve fibers ran parallel to the long axis of root and reached the coronal pulp. They were distributed on Raschkow plexus under the odontoblastic layer. 2. In 2 day group after pulp exposure, tissue necrosis and acute inflammation occurred and CGRP containing nerve fibers increased. In 4 day group, the necrotic tissue extended to the pulp and CGRP containing nerve fibers were distributed around the inflammation zone. 3. In 7 day group after pulp exposure, pulp necrosis occurred, and in 10 day group, the abscess under the necrotic pulp extended to the root apex area and CGRP containing nerve fibers were not observed in root canals. 4.The sprouting of CGRP nerve fibers was most remarkable at the pulp chamber under injury in 4 day group, and it was found at inflammation zone under the necrotic tissue in 7 day group and the remaining root pulp tissue in 10 day group. As mentioned above, CGRP nerve fibers had a tendency to increase around the inflammatory zone, especially around the acute inflammation tissue, when compared with control group. It is suggested that CGRP nerve fibers maybe related to the control of inflammatory response of pulp tissue.

• 대한구강악안면임플란트학회지
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• 제18권2호
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• pp.120-138
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• 2014

Purpose: The purpose of this study was to review the outcomes of a series of studies on tissue regeneration conducted in multiple institutions including the Department of Periodontology, College of Dentistry, Yonsei University. Materials and Methods: Studies were performed divided into the following three subjects; 1) Development of three-dimensional nano-hydroxyapatite (n-HA) scaffold for facilitating drug release and cell adhesion. 2) Synergistic effects of bone marrow-derived mesenchymal stem cells (BMMSC) application simultaneously with platelet-rich plasma (PRP) on HA scaffolds. 3) The efficacy of silk scaffolds coated with n-HA. Also, all results were analyzed by subjects. Results: Hollow hydroxyapatite spherical granules were found to be a useful tool for the drug release and avidin-biotin binding system for cell attachment. Also, BMMSC simultaneously with PRP applied in an animal bone defect model was seen to be more synergistic than in the control group. But, the efficacy of periodontal ligament cells and dental pulp cells with silk scaffolds could not be confirmed in the initial phase of bone healing. Conclusion: The ideal combination of three elements of tissue engineering-scaffolds, cells and signaling molecules could be substantiated due to further investigations with the potentials and limitations of the suggested list of studies.

• 생명과학회지
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• 제28권7호
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• pp.835-840
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• 2018

본 연구는 시토크롬 P450 단백질을 암호화하는 애기장대 유래의 AtCYP78A7을 과발현하는 형질전환 식물체로부터 AtCYP78A7 단백질을 특이적으로 인식하는 단일큰론 항체의 제조와 그 항체를 AtCYP78A7 단백질과 접촉시켜 항원-항체 복합체 형성을 검출함으로써 AtCYP78A7 단백질을 효소면역학적(ELISA) 방법으로 검출하는 진단 키트를 개발하기 위하여 수행하였다. 재조합한 GST-AtCYP78A7 단백질을 항원으로 사용하여 단일클론 항체를 분비하는 융합세포주를 제조한 후 비오틴화 및 페어링 테스트를 통해 포획항체와 검출항체를 선정하였으며, GST-AtCYP78A7 정제 단백질을 기준으로 일품벼, 화영벼, AtCYP78A7 과발현 벼(10B-5, 18A-4)의 용해물을 검출항원으로 사용하여 product test를 진행하였다. 그 결과 AtCYP78A7 단백질에 특이적으로 결합하는 4개의 단클론 항체(mAb 6A7, mAb 4C2, mAb 11H6, mAb 7E8)를 생산하였고, 포획항체 mAb 4C2와 검출항체 mAb 7E8-biotin의 조합으로 ELISA 키트를 개발하였다. 개발된 ELISA 키트를 이용한 벼 시료의 분석 결과 AtCYP78A7 과발현 벼는 전체 단백질 대비 AtCYP78A7 단백질의 비율이 0.1% 이상인 양성으로, 일품벼와 화영벼는 0.1% 미만인 음성으로 나타나 키트를 이용한 AtCYP78A7 단백질의 검출이 가능하였으며, 따라서 본 키트는 향후 AtCYP78A7를 과발현하는 형질전환 작물을 대상으로 하는 환경 모니터링 또는 인체 위해성 평가에 유용하게 활용될 수 있을 것으로 사료된다.

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.100-115
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• 1999

The purpose of this study was to investigate the distribution of calcitonin gene-related peptide containing nerve fibers in rat pulp after dentinl injury by means of immunohistochemistry and confocal laser scanning microscope. The Spague-Dawley rats weighing about 250-300gm were used. The animals were devided into normal control and experimental groups. Experimental animals were sacrified 1, 2, 4, 7, 10, 21days after dentinal injury (dentin cutting, and then acid etching with 35% phosphoric acid) on the maxillary molar teeth. The maxillary teeth and alveolar bone were removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4), then were decalcified with 15% formic acid for 10 days. Serial frozen $50{\mu}m$ thick sections were cut on a cryostat. The rabbit CGRP antibody was used as a primary antibody with a dilution of 1:2000 in 0.01M PB. The sections were incubated for 48 hours at $4^{\circ}C$, and placed into biotinylated antirabbit Ig G as a secondary anti body with dilution of 1:200 in 0.01M PB and incubated in ABC(avidin-biotin complex). The peroxidase reaction was visualized by incubating the sections in 0.05% 3,3 diaminobenzidine tetrahydrochloride containing 0.02% $H_2O_2$. For the confocal laser scanning microscopic examination, Primary antibody reaction was same as immunoperoxidase stainning, but fluorescein isothiocyanate(FITC)-conjugate antirabbit IgG as a secondary antibody was used. The confocal laser scanning microscope was used for the examination. A series of images of optical sections was collected with a 20x objective at $3{\mu}m$ intervals throughout the depth of specimen. FITC fluerescence was registrated through a 488nm and 568nm excitation filter, and images were saved on optical disk. The stereoscopic images and three dimentionnal images were reconstructed by computer software, and then were analyzed. The results were as follows : 1. In normal control group, CGRP containing nerve fibers were coursed through the root with very little branching, and then formed a dense network of terminals in coronal pulp. 2. A slight increase in CGRP containing nerve fibers at 1 and 2day postinjury was noted subjacent to the injury site. In the 4day group, there were an extensive increase in the number of reactive fibers, followed by a partial return toward normal levels at 7~10 day postinjury, and return by 21days. 3. The sprouting of the CGRP containing nerve fibers was evident within 2day after dentinal injury, and by 4days there was a maximal increased, but was decreased at 7days and returned to normal 10~21 day postinjury. 4. In confocal laser scanning microscopic exammination, the distinct distribution pattern and sprouting reaction of CGRP containing nerve fibers were observed in stereoscopic images and three dimentional images. These results suggest that CGRP containing nerve fiber can be important role in the response to dental injury and pain regulation.

• 대한수의학회지
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• 제31권1호
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• pp.71-75
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• 1991

소의 순환혈액 및 림프조직내의 T 및 B 림프구의 분포를 파악하고자 소흉선 세포를 토끼에 과면역시켜 얻은 항혈청을 소의 적혈구, 간장분말 및 골수세포에 순차적으로 흡착시킨 항 흉선 세포혈청(RABTS)과 성숙 T 림프구에만 특이성을 가진 단클론항체 $BLT_1$ 및 B 림프구와 소수의 대식세포에 특이성을 가진 단클론항체 $_6E_{12}$를 1차 항체로 이용하여 ABPC법으로 T 및 B 림프구를 동정한 바 다음과 같은 결과를 얻었다. RABTS 및 $BLT_1$으로 동정한 순환혈액내 단핵세포(PB-MNCs) 중 T림프구의 비율은 각각 70.9${\pm}5.5%$ 및 $59.0{\pm}8.7%$이었으며, PB-MNCs중 nylon wool 비부착세포를 $BLT_1$ 및 $_6E_{12}$로 동정한 T 및 B 림프구의 비율은 각각 $91.6{\pm}1.0%$, $9.6{\pm}0.8%$이었다. 또한 $BLT_1$ 및 $_6E_{12}$로 동정한 서혜림프절, 장간막 림프절, 비장 및 흉선의 T/B림프구의 비율은 각각 $56.4{\pm}6.2/45.3{\pm}7.4%$, $55.6{\pm}7.7/42.3{\pm}5.8%$, $48.6{\pm}5.1/48.5{\pm}6.2%$ 및 $23.0{\pm}4.8/5.6{\pm}2.1%$이었으며, RABTS로 동정한 이들 림프조직내의 T림프구의 비율은 각각 $76.3{\pm}3.4%$, $74.2{\pm}8.2%$, $73.6{\pm}5.5%$ 및 $95.6{\pm}2.8%$이었다. 이상의 결과로 미루어 PB-MNCs네의 T림프구 분리는 nylon wool column법이 보다 효과적임을 알 수 있었다.

• Tuberculosis and Respiratory Diseases
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• 제47권4호
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• pp.478-487
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• 1999

연구배경 : 전자현미경적 관찰에 의해 진단된 흉막의 악성중피종과 전이성선암 환자를 대상으로 하여 7종의 단일클론항체를 이용한 면역조직화학염색을 시행하고 이들의 발현양상을 비교하여 두 질환을 감별할 수 있는 가장 효과적인 염색방법을 알아보고자 하였다. 방 법 : 면역조직화학염색법은 Fisher사(Fisher scientific)의 1-hour immunohistochemistry방법을 이용하였다. 종양조직은 포르말린으로 고정된 파라핀 조직을 이용하여, $3{\mu}m$ 두께로 잘라서, $85^{\circ}C$에서 가열시킨 후 파라핀을 제거하고, 희석액과 단클론항체를 $45^{\circ}C$에서 15분간 반응시켰다. 각 단계마다 automation buffer로 실온에서 세척하였고, avidin-biotin-peroxidase system을 이용한 immunoperoxidase method로 염색하였다. 조직절편은 $45^{\circ}C$에서 10분간 색소원반응을 시행하였고, 대조염색은 hematoxylin으로 실시하였다. 조직 절편은 최소한 각기 다른 두 구역에서 세포질 또는 세포막이 분명하게 염색될 때 양성으로 판정하였다. 결 과 : 7종의 단일클론항체중 CK, EMA, vimentin, S-100 단백과 Leu-M1은 악성중피종과 전이성선암에서 발현율의 차이는 없었으나, B72-3은 전이성선암에서만 발현되었고, CEA는 전이성선암 전례와 악성중피종 42.9%에서 발현되어서 민감도는 높았으나 특이도는 낮았다. 결 론 : B72-3을 이용한 면역조직화학염색은 악성중피종과 전이성선암의 감별진단에 가장 유용한 방법으로 시사되었으며, 기존에 사용되고 있는 CEA와 B72-3을 함께 이용한다면 악성중피종과 전이성선암에 대한 감별진단력을 높일 수 있을 것으로 사료된다.