• Title/Summary/Keyword: Surface marker

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Expression of Hepatitis B Virus S Gene in Pichia pastoris and Application of the Product for Detection of Anti-HBs Antibody

  • Hu, Bo;Liang, Minjian;Hong, Guoqiang;Li, Zhaoxia;Zhu, Zhenyu;Li, Lin
    • BMB Reports
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    • v.38 no.6
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    • pp.683-689
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    • 2005
  • Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.

A study on image registration and fusion of MRI and SPECT/PET (뇌의 단일 광자 방출 전산화 단층촬영 영상, 양전자 방출 단층 촬영 영상 그리고 핵자기공명 영상의 융합과 등록에 관한 연구)

  • Joo, Ra-Hyung;Choi, Yong;Kwon, Soo-Il;Heo, Soo-Jin
    • Progress in Medical Physics
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    • v.9 no.1
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    • pp.47-53
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    • 1998
  • Nuclear Medicine Images have comparatively poor spatial resolution, making it difficult to relate the functional information which they contain to precise anatomical structures. Anatomical structures useful in the interpretation of SPECT /PET Images were radiolabelled. PET/SPECT Images Provide functional information, whereas MRI mainly demonstrate morphology and anatomical. Fusion or Image Registration improves the information obtained by correlating images from various modalities. Brain Scan were studied on one or more occations using MRI and SPECT. The data were aligned using a point pair methods and surface matching. SPECT and MR Images was tested using a three dimensional water fillable Hoffman Brain Phantom with small marker and PET and MR Image was tested using a patient data. Registration of SPECT and MR Images is feasible and allows more accurate anatomic assessment of sites of abnormal uptake in radiolabeled studies. Point based registration was accurate and easily implemented three dimensional registration of multimodality data set for fusion of clinical anatomic and functional imaging modalities. Accuracy of a surface matching algorithm and homologous feature pair matching for three dimensional image registration of Single Photon Emission Computed Tomography Emission Computed Tomography (SPECT), Positron Emission Tomography (PET) and Magnetic Resonance Images(MRD was tested using a three dimensional water fill able brain phantom and Patients data. Transformation parameter for translation and scaling were determined by homologous feature point pair to match each SPECT and PET scan with MR images.

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Ultrasound Imaging of Normal Cardiac Structures with Convex Scanner in Puppies (강아지에서 컨벡스스캐너를 이용한 정상적인 심장구조물의 초음파상)

  • Jeong Soon-wuk;Park Hee-myung;Han Sook-hee;Yoon Jung-hee;Han Hong-ryul
    • Journal of Veterinary Clinics
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    • v.11 no.2
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    • pp.529-537
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    • 1994
  • Sector scanner which has a conical end is used to image through the intercostal space because heart is protected by the ribs. Cardiac data published all around the world were also obtained by sector scanner. Although scanners being used in every small animal practice and animal hospital at college in Korea include convex ape and linear type, linear type is not appropriate f3r cardiac scan because of a wide contact surface. The purpose of this study is to establish ultrasonographic images of normal cardiac structures by measuring shape, size of reflectable cardiac structure according to restraint position in scanning normal heart of the puppies with 6.5 MHz convex scanner(SonoAce 4500, Medison, Korea) used in our veterinary teaching hospital, Seoul national university. Seventeen male and female puppies considered having healthy hear by X-ray and clinical examination are used feom April to July 1994. Scanning point selection of probe head and the distinction of imaged cardiac structures were accomplished by necropsy and cardiac scanning performed through thoracotomy under general anesthesia. At 10 o'clock position of transducer(at an angle of 30$^{\circ}$ between imaginary line from elbow joint to 3rd sternum and probe head, 60$^{\circ}$ from body surface, 4th intercostal space of right thorax) with the marker of scanner toward the head of dogs right atrium, left atrium and left ventricle were observed in 2, 3, 4, 5 intercostal space(2cm from the sternum) of experimental dog positioned ventrodorsally under general anesthesia. Under these conditions, the numerical values of imaged diastolic hear are as follows : the distance from skin to apex(mean$\pm$S.D) 47.53$\pm$6.94mm, thickness of left ventricular wall 6.00$\pm$1.60mm, length of left ventricle 16.27$\pm$5.31mm, width of left ventricle 15,33$\pm$4.25mm, length of left atrium 12.33$\pm$3.82mm, width of left atrium 11. 33$\pm$3.94mm, length of right atrium 1.00$\pm$2.41mm, width of right atrium 11.21$\pm$2.76mm and the area of left ventricle 270.92$\pm$109.81mm$^2$, area of left atrium 98.00$\pm$41.08mm$^2$, area of right atrium 62.75$\pm$21.04mm$^2$.

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Immunomodulatory activities of crude polysaccharide fraction separated from Perilla frutescens Britton var. acuta Kudo (자소엽(Perilla frutescens Britton var. acuta Kudo) 조다당류 추출물의 면역활성 효과)

  • Byun, Eui-Hong
    • Korean Journal of Food Science and Technology
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    • v.49 no.5
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    • pp.559-566
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    • 2017
  • This aim of this study was to examine the immunomodulatory activities of crude polysaccharides from Perilla frutescens Britton var. acuta Kudo (PCP) in mouse bone marrow-derived dendritic cells (BMDC) and splenocytes. The immunomodulatory activity was determined by cell viability, nitric oxide (NO) production, cell surface marker expression (CD 80/86 and MHC class I/II), and cytokine production in BMDC, and cell viability, and cytokine production in splenocytes. Cell proliferation and cytokine production (tumor necrosis factor; TNF-${\alpha}$, interleukin (IL)-6, IL-$1{\beta}$, and IL-12) tested in BMDC were significantly increased by PCP treatment. Additionally, the cell surface markers (CD 80/86, MHC class I/II) were highly increased by PCP treatment. For cytokine production in splenocytes, PCP treatment significantly increased the production of Th 1 cytokines [IL-2 and interferon (IFN)-${\gamma}$], but not Th 2 cytokines (IL-4). Therefore, PCP can induce immune cell activation and is a potential candidate for the development of nutraceuticals to boost the immune system.

Establishment of Human Embryonic Stem Cells using Mouse Embryonic Fibroblasts and Human Fetal Fibroblasts as Feeder Cells (인간태아 섬유아세포와 생쥐배아 섬유아세포를 기저세포로 활용한 인간 배아줄기세포의 확립)

  • Cho, Hye Won;Ko, Kyoung Rae;Kim, Mi Kyoung;Lee, Jae Ik;Sin, Su Il;Lee, Dong Hyung;Kim, Ki Hyung;Lee, Kyu Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.133-147
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    • 2005
  • Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.

Establishment of In Vitro 3-Dimensional Culture System of Mouse Endometrial Cells;I. Cytohistological Study on Mouse Endometrium (마우스 자궁내막 세포를 이용한 3차원적 배양시스템 확립에 관한 연구; I. 마우스 자궁내막에 관한 세포조직학적 연구)

  • Nam, Hwa-Kyung;Kim, Eun-Young;Lee, Keum-Sil;Park, Sae-Young;Park, Eun-Mi;Kwon, Jung-Kyun;Yoon, San-Hyun;Park, Se-Pil;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.31-37
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    • 2000
  • This study was designed to identify the ultrastructural changes of mouse endometrium during peri-implantation period and obtain the fundamental information for the establishment of 3-dimensional culture system of mouse endometrial cells in vitro. The used female ICR mice ($6{\sim}8$ wks) were conducted on pregnant. The biopsies were obtained from whole uterus at cycle day 1 (D1) and day 5 (D5) after hCG injection and mating. The biopsies materials were fixed 2.5% glutaraldehyde and 1% osmium tetroxide. Subsequently, for observation using light and transmission electron microscopy (LM and TEM), they were dehydrated and embedded in Epon and the embedded biopsies were sectioned and stained. For scanning electron microscopy (SEM), the fixed specimens were dehydrated, dried and coated with gold. 1) For LM, the biopsied materials at D5 (late secretory phase) were appeared the extended stromal layer by increased connective tissues and the fully developed endometrial glands and vessels compared with D1 (early secretory phase). 2) For TEM, the mouse endometrium was consisted of 3-layers, a simple polarized columnar epithelial cells, basement membrane and stromal cells. At D5, the distribution of microvilli, endoplasmic reticulum, Golgi body, lipid and glycogen deposits, secretory granules and surface area of basement membrane were increased. 3) For SEM, the degree of folding and microvilli of surface of mouse epithelial cells was became more and more according to the process of secretory phase, and at D5, implantation time of mouse, the appearance of pinopodes as a specific marker of uterine receptivity was found. The uterine pinopodes of mouse were found in narrow sites at the luminal surface, irregularity and appeared the different stages in the same sample. Therefore, these results indicated that the mouse endometrium was experienced dramatic morphological changes during peri-implantation period.

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The Effect on the Lower Limbs Joint as the Landing Height and Floor Pattern (착지 높이와 지면 형태가 하지 관절에 미치는 영향)

  • Kim, Eun-Kyong
    • Korean Journal of Applied Biomechanics
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    • v.21 no.4
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    • pp.437-447
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    • 2011
  • In this study, the lower limbs joints were analyzed for features based on the biomechanical characteristics of landing techniques according to height and landing on the ground type (flats and downhill). In order to achieve the objectives of the study, changes were analyzed in detail contents such as the height and form of the first landing on the ground at different angles of joints, torso and legs, torso and legs of the difference in the range of angular motion of the joint, the maximum angular difference between joints, the lower limbs joints difference between the maximum moment and the difference between COM changes. The subjects in this study do not last six months did not experience joint injuries 10 males in 20 aged were tested. Experimental tools to analyze were the recording and video equipment. Samsung's SCH-650A model camera was used six units, and the 2 GRF-based AMTI were used BP400800 model. 6-unit-camera synchronized with LED (photo cell) and Line Lock system were used. the output from the camera and the ground reaction force based on the data to synchronize A/D Syc. box was used. To calculate the coordinates of three-dimensional space, $1m{\times}3m{\times}2m$ (X, Y, Z axis) to the size of the control points attached to the framework of 36 markers were used, and 29 where the body was taken by attaching a marker to the surface. Two kinds of land condition, 40cm and 60cm in height, and ground conditions in the form of two kinds of flat and downhill slopes ($10^{\circ}$) of the landing operation was performed and each subject's 3 mean two-way RM ANOVA in SPSS 18.0 was used and this time, all the significant level was set at a=.05. Consequently, analyzing the landing technique as land form and land on the ground, the changes of external environmental factors, and the lower limbs joints' function in the evaluation were significantly different from the slopes. Landing of the slop plane were more load on the joints than landing of plane. Especially, knee extensor moment compared to the two kinds of landing, slopes plane were approximately two times higher than flat plane, and it was statistical significance. Most of all not so much range of motion and angular velocity of the shock to reduce stress was important. In the further research, front landing as well as various direction of motion of kinetic, kinetic factors and EMG variables on lower limbs joints of the study in terms of injury-prevention-approach is going to be needed.

Identification of CM1 as a Pathogenic Factor in Inflammatory Diseases and Cancer

  • Bae, Se-Yeon;Kim, Hyem-In;Yu, Yeon-Sil;Lee, Na-Eun;Kong, Joo-Myoung;Kim, Hang-Rae;Hwang, Young-Il;Song, Yeong-Wook;Kang, Jae-Seung;Lee, Wang-Jae
    • IMMUNE NETWORK
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    • v.11 no.3
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    • pp.175-181
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    • 2011
  • Background: CM1 (centrocyte/-blast marker 1) was defined by a mAb against concavabalin-A (ConA) activated PBMC. It is expressed in germinal center of human tonsil and on the surface of activated PBMC as well as cancer cells. Recently, increased productions of pro-inflammatory mediators were detected from activated PBMC by CM1 ligation. Methods: However, there is a limitation to explain the exact role of CM1 on inflammation and its related mechanisms, since the identity of CM1 is still not clarified. In our previous study, we have already confirmed that soluble form of CM1 was produced by Raji. Therefore, we performed Q-TOF analysis after immunoprecipitation of concentrated Raji culture supernatant using anti-CM1 mAbs. Results: As a result, we found that CM1 is identical to enolase-1(ENO1), a glycolytic enzyme, and we confirmed that results by silencing ENO1 using siRNA. It was also confirmed through competition assay between anti-CM1 and anti-ENO1 mAbs. Finally, we investigated the possible role of CM1 in inflammatory response and cancer. The ligation of CM1 on Raji cells with anti-CM1 mAbs induces the extensive production of prostaglandin $E_2(PGE_2)$. In addition, the increased activity of matrix metalloproteinase (MMP)-2/9 was shown in NCI-N87, stomach cancer cell line by CM1 stimulation. Conclusion: CM1 is identical to ENO1 and it might be an important role in the regulation of inflammatory responses.

THE ONSET OF ANKYLOSIS FOLLOWING INTRUSIVE LUXATION INJURIES (외상성 intrusion 치아의 교정적 견인시기에 관한 실험적 연구)

  • Chung, Kyu-Rhim;Turley, Patrick-K.
    • The korean journal of orthodontics
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    • v.21 no.2 s.34
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    • pp.259-272
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    • 1991
  • Orthodontic traction has been suggested as the treatment of choice for intrusive luxation injuries. Prior research has shown orthodontic forces to be ineffective in the presence of ankylosis or in cases with zero mobility following the injury. If orthodontic traction is to be effective, it must be initiated prior to the onset of ankylosis. The purpose of this study was to describe the effects of intrusive luxation at various times following the injury, and to determine the time of the onset of ankylosis, and to examine what effect immediate partial luxation has on the onset of ankylosis. Eight young mongrel dogs were utilized for this study. Intrusive luxation was produced with an axial impact using a gravity hammer and a specially designed holding device on 4 teeth (2 max. and 2 man. first premolars) in each dog. The teeth were intruded approximately 3-4mm in an axial direction. One maxillary and one mandibular premolars were partially luxated with the other two teeth being untouched. Pre and posttrauma tooth position was documented with plaster models and radiographs taken with an individualized X-ray jig. Dogs were sacrificed immediately following the injury and at 1, 2, 4, 7, 10, 14 and 21 days respectively. Tetracycline was administered as a vital bone marker 24 hours before sacrifice. Block sections of the tooth and alveolus were prepared for decalcified and non decalcified histologic sections. The effects of traumatic intrusion were analyzed by means of model casts, radiographs, tetracycline bone marking and histologic preparations. The results obtained were as follows: 1. The animal sacrificed immediately following the injury displayed alveolar fractures, torn periodontal ligaments, and areas of direct tooth-bone contact. 2. The odontoblastic layer of the pulp was disorganized as early as 24 hours after the injury. 3. Bony remodeling was noted at 4 days along with active surface resorption. 4. Ankylosis was first seen 7 days after the injury. 5. Osteogenesis in the dentin (thick tetracycline bands) was observed 7 days after the injury. 6. There was no progressive root resorption and ankylosis where the periodontal ligament has been healed. 7. The Luxated group showed significantly more root resolution and ankylosis than the Nonluxated group with increased observation periods. The results suggest that ankylosis may occur within the first week following the injury, and hence orthodontic traction should be initiated as soon after the injury as possible.

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Immunological Synergistic Effects of Combined Treatment with Herbal Preparation (HemoHIM) and Red Ginseng Extracts (마우스세포를 이용한 홍삼추출물과 생약복합추출물의 병용 처리에 따른 면역활성 효과)

  • Byun, Myung-Woo;Byun, Eui-Hong
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.2
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    • pp.182-190
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    • 2015
  • This present study demonstrates the immunological synergistic effects of herbal preparation (HemoHIM) and red ginseng powder granule in various immune cell models (bone marrow-derived macrophages, dendritic cells, and mouse splenocytes) from mice. Both herbal preparation and red ginseng extracts were treated to bone-marrow derived macrophages, dendritic cells, and mouse splenocytes, and there was no cytotoxicity at a dose below $200{\mu}g/mL$. Cell proliferation and cytokine [tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-6, and IL-12] production tested in bone marrow-derived macrophages and dendritic cells significantly increased upon combined treatment. Cell surface marker (CD 80/86, MHC class I/II)-mediated immune cell activation was highly elevated by combined treatment. For cytokine production in splenocytes, combined treatment significantly increased production of Th 1 type cytokines [IL-2 and interferon (IFN)-${\gamma}$] but not Th 2 type cytokines (IL-4 and IL-10). Therefore, combined treatment with HemoHIM and red ginseng extracts is an effective method to establish powerful immunological synergy in immune cells.