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Effects of Scutellariae Radix on Neuronal Apoptosis of Spinal Cord Contusion Injury in Rats (황금(黃芩)이 척수압박손상 흰쥐의 척수신경세포 자연사에 미치는 영향)

  • Bahn, Hyo-Jung;Jo, Jong-Jin;Kim, Bum-Hoi;Park, Seong-Ha;Shin, Jung-Won;Kim, Seong-Joon;Sohn, Nak-Won
    • Journal of Korean Medicine Rehabilitation
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    • v.21 no.4
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    • pp.13-22
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    • 2011
  • Objectives : This Study was performed to evaluate the effects of Scutellariae Radix(SR) water-extract on the tissue and neuronal apoptosis of the spinal cord injury(SCI). Methods: SCI was induced by mechanical contusion following laminectomy of 10th thoracic vertebra in Sprague-Dawley rats. SR was orally given once a day for 7 days after SCI. Neuronal apoptosis was examined with terminal deoxynucleotidyl transferase-mediated dUTPnick-end labeling(TUNEL) assay. Bax (Bcl-2-asociated X protein), Bcl-2(B-cell blastoma 2), c-Fos(FBJ osteosarcoma oncogene) expressions were examined using immuno-histochemistry. Individual TUNEL and immuno-labeled cells expressing Bax, Bcl-2 and c-Fos were counted on the same level in peri-damaged region and in ventral horn. Results: 1. SR significantly reduced number of TUNEL labeled apoptotic cells induced by the spinal cord contusion injury. 2. SR significantly reduced Bax positive cells expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. 3. SR strengthened Bcl-2 expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. 4. SR reduced c-Fos expression on the motor neuron in the ventral horn induced by the spinal cord contusion injury. Conclusions : These results suggest that SR plays an inhibitory role against neuronal apoptosis and has significant effects for locomotor disfunction induced by SCI.

The Effects of Bone Morphogenetic Protein-4 and Resorbable Membrane on the Regeneration of Periodontal Tissues (골형태형성단백질과 흡수성차폐막이 치주조직 재생에 미치는 영향)

  • Lim, Sang-Cheol;Kwon, Young-Hyuk;Lee, Man-Sup;Partk, Joon-Bong
    • Journal of Periodontal and Implant Science
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    • v.30 no.4
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    • pp.757-779
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    • 2000
  • The aim of our study is to achieve complete periodontal tissue regeneration by the application of BMP and resorbable membrane. Three beagle dogs aged over one and half years and weighed 14 to 16 kg were used in this study. Mandibular 1st, 2nd premolars were extracted bilaterally. Horizontal furcation defects were induced around 3rd, 4th premolars bilaterally. BMP-4 were applied in the right side with resorbable membranes and only resorbable membranes were applied in the left side respectively. Each animal was sacrificed at 2, 4, and 8weeks, after regenerative surgery. Specimens were prepared with Hematoxylin-Eosin stain and Goldner's modified Masson Trichrome stain for light microscopic evaluation. The results were as follows: 1. At 2 weeks after regenerative surgery, downgrowth of junctional epithelium was observed both in the membraneapplied site and BMP-4-and-membrane-applied site. 2. At 4 weeks after regenerative surgery, resorbable membranes were completely resolved, therefore would not prevent downgrowth of junctional epithelium. New bone formation, new cementum formation and Sharpey's fiber were observed in BMP-4-andmembrane-applied site. 3. At 8 weeks after regenerative surgery, downgrowth of junctional epithelium was observed in the membrane-applied site. But, new cementum formation was observed in the same site. The extensive regeneration of new bone, new cementum and remarkable formation of Shapey's fiber were showed in BMP-4-and-membrane-applied site. 4. Resorbable membranes were resolved via the cell-mediated processes. 5. Periodontal tissue regeneration were better achieved in the BMP-4-andmembrane-applied site than in the membrane-applied site. Within the above results, BMP-4 may have the strong capability to form the new bone and resorbable membrane may be able to prevent the bony ankylosis. However, resolution rate of resorbable membrane may not be enough to protect rapid epithelial downgrowth for ideal periodontal regeneration. In conclusion, I suggest BMP-4 may have the strong possibility to be utilized in the clinical periodontal treat-

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Changes of the Cellularities in the Bronchoalveolar Lavage Fluid of the Experimental Silicosis (흰쥐규폐증에서 기관지 폐포 세척액내 세포변화)

  • Lim, Young;Yun, Im-Goung;Oh, Min-Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.38 no.2
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    • pp.143-154
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    • 1991
  • Pneumconiosis is a sort of pulmonary fibrosis consequent to the inhalation of the respirable dusts. Thus, the pathogenesis of silicosis have concentrated largely on the early response of alveolar macrophage and the later fibroblastic stimulation. But the role of the other cells and continuing cell injury in the pathogenesis has not been fully studied. And the chemical factors such as prostaglandin, fibroblast stimulating factor and inhibiting factor and chemotaxin are also participated in the mechanism of pulmonary fibrosis in silicosis. In order to clarify the role of alveolar cells and prostaglandin, we investigated the changes of the cellularities in bronchoalveolar lavage fluid and tissue pathology in the experimental silicosis with the time sequence. The experimental animals were divided into 3 groups; control group received only intratracheal injection of 0.5 ml saline, silica group received the intratracheal instillation of 40 mg silica with the same amount saline, and aspirin group received 450 mg/kg of aspirin after silica instillation. The results were as follows: 1) The total cells of bronchoalveolar lavage fluid in the silica group markedly increased in comparison with the control group, but there was no significant difference between the silica and aspirin groups. 2) The percentages of alveolar macrophages to the total number of cells in the silica group tended to be lower than those in the control group and also lower than those in the aspirin group at the 1st week after silica instillation. 3) The percentages of neutrophils to the total number of cells in the silica group were significantly higher than those in the control group during the entire period and also higher than those in the aspirin group at the 3rd day after silica instillation. 4) In the silica group, the percentages of lymphocytes to the total number of cells were increased 143 progressively with the time course and those were significantly higher than those in the control group from the 3rd week after silica administration. There were marked differences of lymphocyte percentages between the silica and aspirin groups at the 1st week after silica instillation. 5) The inflammatory change was observed in the rat lung at the 1st day after silica instillation. Also the silicotic nodule appeared in the silica group at the 1st week but we could not find out that nodule in the aspirin group at that time. The fibrotic changes in the rat lung tended to be increased progressively with the time course, therefore, the diffuse fibrotic pattern appeared in the whole field at the 20th week after silica instillation. 6) By the electron microscopy, there were gradual increases of phagosomes and vacuoles in the alveolar macrophage in the silica group as compared with the control group. These results suggest that the neutrophils and the lymphocytes have also participated in the pulmonary fibrosis even though the alveolar macrophage has a major role, and prostaglandin mediate the inflammation and pulmanary fibrosis in the experimental silicosis.

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A study of the clinical effects of various bone graft materials (다양한 골 이식재의 임상 효과에 대한 고찰)

  • Lee, Seung-Bum;Yon, Je-Young;Chae, Gyung-Joon;Jung, Ui-Won;Kim, Chang-Sung;Lee, Yong-Geun;Cho, Kyoo-Sung;Chai, Jung-Kiu;Kim, Chong-Kwan;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.37 no.4
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    • pp.719-732
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    • 2007
  • Purpose: Various bone graft materials are being used for periodontal tissue regeneration. Th materials are being developed continuously for ideal clinical effects. Therefore, it is necessary to identify the clinical characteristics of each bone graft material through comparing the various bone graft materials statistically and in doing so, proposing a more efficient bone graft material. In this study, the following results were attained through comparing the clinical effects among the bone graft materials, using the statistical method based on the clinical studies published at the department of periodontology of Yonsei hospital. Materials and Method: 6 selected studies of department of Periodontology at Yonsei University Hospital were based on clinical study of bone grafting in intrabony defects. It was compared the clinical parameters among the 6 clinical studies, using the statistical META analysis. Result: When comparing the probing depth reduction, there was a relatively great amount of decease when using the xenograft, Anorganic Bovine Derived Hydroxapatite Bone Matrix/Cell Binding Peptide(ABM/P-15: PepGen $P-15^{(R)}$) and the autogenous bone and absorbable membrane, d, 1-alctide/glycolide copolymer(GC: $Biomesh^{(R)}$). The allogfrafts showed a relatively low decrease in the probing depth and clinical attachment change. It also showed a slight decrease in the bone probing depth. The allografts showed various results according to different bone graft materials. When comparing the ABM/P-15 and bovine bone $powder(BBP^{(R)})$, ABM/P-15 showed a relatively high clinical attachment level and the bovine bone powder showed a relatively high clinical attachment level. The probing depth change and gingival recession change showed a lower value than the mean value between the two bone graft materials. The synthetic bone showed a relatively high decrease in clinical attachment level and periodontal probing depth change. There was a relatively larger amount of gingival recession when using Bioactive Glass(BG) but a relatively low bone regeneration effect was seen. Conclusion: Good restorative results of the periodontal tissue can be attained by applying the various bone graft materials being used today after identifying the accurate clinical effects.

In Vitro Isolation and Proliferation of Mouse Male Germ-Line Stem Cells (생쥐 생식줄기세포의 체외 분리 및 증식)

  • 김수경;김계성
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.243-248
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    • 2003
  • Sperrnatogenesis, the process by which the male germ-line stem cells(GSCs; type A spermatogonia) divide and differentiate to produce the mature spermatozoa, occurs in the seminiferous tubules of the testis. The GSCs proliferate actively to produce two types of cells: other GSCs and differentiating spermatogonia. GSCs have unipotentcy, devoted solely to the generation of sperm. The function of GSCs has broad implications for development, disease, and evolution. Spermatogenesis is fundamental for propagation of species and the defects of this system can result in infertility or disease. The ability to identify, isolate, culture, and alter GSCs will allow powerful new approaches in animal transgenesis and human gene therapy relating to infertility. Until recently, research on stem cells in the testis has been limited because of technical difficulties in isolating and identifying these cell populations. Here, we were trying to find out optimal conditions for in vitro culture of GSCs for identifying and isolating GSCs. We collected mouse GSCs from 3-days old mouse by two-step enzyme digestion method. GSCs were plated and grown on mouse embryonic fibroblasts in Dulbecco's modified Eagle's medium (DMEM) containing 15% fatal bovine serum, 10 mM 2-mercaptoethanol, 1% non-essential amino acids, 1 ng/$m\ell$ bFGF, 10 $\mu$M forskolin, 1500 U/$m\ell$ human recombinant leukemia inhibitory factor (LIF). Over a period 3∼5 days, GSCs gave rise to large multicellular colonies resembling those of mouse pluripotent stem cells. After 5th passages, cells within the colonies continued to be alkaline phosphatase and Oct-4 positive and tested positive against a panel of two immunological markers(Integrin $\alpha$ 6 and Integrin $\beta$ 1) that have been recognized generally to characterize GSCs. SSEA-1, SSEA-3, and SSEA-4 also showed positive signals. Based on our data, these GSCs-derived cultures meet the criteria for GSCs itself and even other pluripotent stem cells. We reported here the establishment of in vitro cultures from mouse male GSCs.

Nucleotide sequence analysis and expression of NSP4 gene of human rotaviruses isolated in Korea (국내에서 분리된 사람 로타바이러스의 NSP4 유전자 염기서열 분석 및 발현)

  • Jung, Dong-hyuk;Song, Yun-kyung;Kim, Kyung-mi;Park, Hyo-sun;Back, Myoung-soon;Kang, Shien-young
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.89-100
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    • 2002
  • The nonstructural glycoprotein NSP4, encoded by the 10th gene of rotavirus, has been known to play important roles in viral assembly and pathogenesis. The NSP4 genes of human rotavirus Korean isolates, designated as CBNU/HR-1, CBNU/HR-2, CBNU/HR-3, and CBNU/HR-4, were cloned, sequenced and characterized. Also, the NSP4 gene of the CBNU/HR-1 was expressed in a baculovirus-insect cell system. The sequence data indicated that the NSP4 genes of human rotavirus Korean isolates were 750 or 751 bases in length and encoded one open reading frame of 175 amino acids. Two glycosylation sites were recognized in the NSP4 gene of human rotavirus isolates tested. The NSP4 of CBNU/HR-1, CBNU/HR-3, and CBNU/HR-4 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype B viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype A viruses. However, the NSP4 of CBNU/HR-2 exhibited a high degree of amino acid sequence homology with that of NSP4 genotype A viruses, but a low degree of amino acid sequence homology with that of NSP4 genotype B viruses. The Sf9 cells infected with recombinant baculovirus, inserted with NSP4 gene of CBNU/HR-1, produced specific cytopathic effects and the expressed NSP4 was detected by immunofluorescence staining using NSP4-specific monoclonal antibody(MAb). The expressed NSP4 migrated at 16-26 kDa on SDS-PAGE and reacted with NSP4-specific MAb by Western blotting.

Respiratory Protective Effect of Salvia plebeia R. Br. Extracts against Ambient Particulate Matter-induced Airway Inflammation (미세먼지 유도 기도염증에 대한 배암차즈기 추출물의 호흡기 보호 효과)

  • Song, Hyeongwoo;Ji, Kon Young;Kim, Bok Kyu;Yang, Won Kyung;Han, Chang Kyun;Shin, Han Jae;Park, Yang Chun;Hwang, Ji Sook;Kang, Hyung Sik;Kim, Seung Hyung
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.5
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    • pp.269-281
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    • 2017
  • Background: Small particles increase airway inflammation upon reaching the alveoli. Here, we investigated the protective or therapeutic effects of Salvia plebeia R. Br. (SP_R) extracts on airway inflammation. Methods and Results: To investigate the anti-inflammatory activity of SP_R extracts, we measured their inhibitory effect on the production of reactive oxygen species (ROS) expression of inflammatory mediators, and immune cell infiltration in MH-S alveolar macrophage cells and in the ambient particulate matter (APM)-exposed airway inflammation mice model. The SP_R extracts inhibited the production of ROS and expression of IL-4, IL-10, IL-15, and IL-17A mRNA in APM-stimulated MH-S cells. Oral administration of SP_R extracts suppressed APM-induced inflammatory symptoms, such as high alveolar wall thickness, excess collagen fibers, decreased mRNA expression of chemokines (Ccr9, Ccl5, Ccr3), inflammatory cytokines (IL-15, TNF-${\alpha}$), and IL-4 Th2 cytokine in the lung. The SP_R extracts also inhibited ROS production, granulocyte ($CD11b^+Gr-1^+$) infiltration, IL-17A, TNF-${\alpha}$, macrophage inflammatory protein (Mip-2), and chemokine (C-X-C motif) ligand 1 (Cxcl-1) production in the airway. The specific compounds in the SR-R extracts that mediate the anti-inflammatory effects were identified. Conclusions: In this study, SP_R extracts effectively inhibited airway inflammatory responses, such as ROS production and granulocyte infiltration into the airway, by regulating the expression of chemokines and inflammatory cytokines.

Isolation and identification of Cyuptosporidium from various animals in Korea (한국산 동물로부터 크립토스포리디움의 분리 및 동정 III. 닭으로부터 Cryptosporidium baileyi의 분리)

  • Lee, Jae-Gu;Seo, Yeong-Seok;Park, Bae-Geun
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.315-324
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    • 1991
  • Each of SPF chicken(Hi-Line strain, 2-day-old males) was inoculated with 2.5 or $5\times10^4$ oocysts by stomach tube. The oocyst was the medium type of Cryptosporidium previously isolated from Korean chicken origin, and passed in 2-day-old SPF chicken. The patterns of oocyst discharge were monitored daily, and in order to observe the ultrastructure of the developmental stages, the bursa of Fabricius of the chicken was examined by transmission electron microscopy (TEM) on the 12th day postinoculation. The prepatent period for 8 chicken was 5.9 days postinoculation on the average, and the patent period was 12.9 days. The number of oocysts discharged per day for the chicken was reached peak on day 12 postinoculation on the average. A large number of oocysts was found in fecal samples obtained from inoculated chicken on days 8~14 postinoculation. The ultrastructural feature of almost every developmental stage of the medium type from chicken was very similar to that of Cryptosporidium previously isolated from mammalia including human and birds except for the attachment site of C. tsuris to the mucus cell from mammalia, but dimension of the oocysts from fecal samples of the medium type was different from those of C. meleagridis and mammalia origin. The above results reveal that the medium type of Cryptosporidium of Korean chicken origin is identified as Cryptosporidium baileyi.

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The Effect of the Unsaturated Oil on the Normal Liver and Lipid Metabolism of Rats Fed Several Plant Oils (불포화도가 다른 식물성 유지를 섭취시킨 흰쥐에서 정상적 간 및 지질대사변화의 고찰)

  • 서화중;김선희;정두례
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.5
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    • pp.426-432
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    • 1991
  • To investigate the effect of the unsaturated plant oil on the normal liver and lipid metabalism rats were fed the daily dose of 1.57ml/kg body weight of perilla oil (Iodine value 190~207), corn oil (Iodine value 115~130) and olive oil (Iodine value 80~85) respectively for 28 days. The results were as follows. For the 14 days the test groups showed normal weight gain with 7.86~20.89% increase rate. In the period of the 3rd and the 4th week the increase rate of the perilla oil group was decreased significantly under 17.53~13.8% of control level, but the remainders(corn oil, olive oil) keep normal. The feeding of corn and olive oil for 28 days did show any harmful effect on normal GOT, GPT activity, ALK-P, serum cholesterol and serum triglyceride value of rat. The perilla oil feeding for 21~28 days slightly increased the GOT, GPT activity as 174.87, 93.46u but decreased the cholesterol and triglycerids value as 54.6~0.36mg/dl compared to control. In the pathological finding of test group liver some rats in 28 days feeding group showed reactive vesicula nuclei in corn oil group and mild fatty metamorphosis in olive oil group. But most subjects did not show any characterized sign of acute or subacute liver damage.

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Expression Study of The Mouse Collectin-Placenta 1 Gene (마우스 Collectin-Placenta 1 유전자의 발현 연구)

  • Kim, Geun Ho;Kim, Youn Uck
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.8
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    • pp.477-484
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    • 2019
  • Several types of scavenger receptors, including the Collectin-Placenta 1 (CL-P1) receptorthat is present in mammals, are molecules that are expressed on the surfaces of endothelial cells, macrophages and smooth muscle cells. These molecules are cell surface glycoproteins that can be conjugated to oxidized low density lipoprotein (oxLDL). Among these molecules, the effect of quercetin on CL-P1 activation has been confirmed. Quercetin is known as an antioxidant that stops oxidation because it acts to remove free radicals that are responsible for the oxidation reaction. In this study, fragments from the transcription start site of the mouse CL-P1 gene promoter to the -500th base were cloned using DNA polymerase. These fragments were then introduced into macrophage like RAW264.7 cells and fibroblast-like NIH3T3 cells to study the effect of quercetin on the CL-P1 gene expression. As a result, we found that bases ranging from -250 to -350 in the anterior part where gene expression starts are important for producing CL-P1 protein. Among them, the DNA mutation experiments we performed confirmed that the E2F binding sites are critical for producing the CL-P1 protein? In addition, when quercetin was added to the RAW264.7 culture medium, which was a culture of adherent cells, observedthe phenomenon of the cells falling off from the surface of the culture container.