• The Korean Journal of Physiology and Pharmacology
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• 제26권4호
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• pp.287-295
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• 2022

Staphylococcus aureus (S. aureus) is known to induce apoptosis of host immune cells and impair phagocytic clearance, thereby being pivotal in the pathogenesis of atopic dermatitis (AD). Adipose-derived stem cells (ASCs) exert therapeutic effects against inflammatory and immune diseases. In the present study, we investigated whether systemic administration of ASCs restores the phagocytic activity of peripheral blood mononuclear cells (PBMCs) and decolonizes cutaneous S. aureus under AD conditions. AD was induced by injecting capsaicin into neonatal rat pups. ASCs were extracted from the subcutaneous adipose tissues of naïve rats and administered to AD rats once a week for a month. Systemic administration of ASCs ameliorated AD-like symptoms, such as dermatitis scores, serum IgE, IFN-γ⁺/IL-4⁺ cell ratio, and skin colonization by S. aureus in AD rats. Increased FasL mRNA and annexin V⁺/7-AAD⁺ cells in the PBMCs obtained from AD rats were drastically reversed when co-cultured with ASCs. In contrast, both PBMCs and CD163⁺ cells bearing fluorescent zymosan particles significantly increased in AD rats treated with ASCs. Additionally, the administration of ASCs led to an increase in the mRNA levels of antimicrobial peptides, such as cathelicidin and β-defensin, in the skin of AD rats. Our results demonstrate that systemic administration of ASCs led to decolonization of S. aureus by attenuating apoptosis of immune cells in addition to restoring phagocytic activity. This contributes to the improvement of skin conditions in AD rats. Therefore, administration of ASCs may be helpful in the treatment of patients with intractable AD.

• 한국임상수의학회지
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• 제40권1호
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• pp.1-7
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• 2023

The growing market for companion animals, combined with their increasing lifespan, has generated an increased interest in companion animal immunity enhancers. Ginsenoside, a saponin component of ginseng and an essential ingredient of red ginseng marc (produced during red ginseng production), is effective in improving immunity. In this experiment, a powder mixture of red ginseng marc and steamed red ginseng extract powder (RGME) was orally administered to dogs for eight weeks. Subsequently, blood samples were collected and tested every four weeks. In addition, canine peripheral blood mononuclear cells (cPBMCs) were stimulated with or without interleukin-2 (IL-2) to evaluate their proliferation and cytokine secretion abilities. Proliferation assay suggests that the administration of RGME effectively enhanced numbers of cPBMCs under IL-2 stimulation. Furthermore, in the RGME group, a significant increase in the concentration of interferon gamma released from cPBMCs under IL-2 stimulation was observed. In conclusion, RGME might be an effective health supplement for improving immunity in dogs.

• Journal of Veterinary Science
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• 제24권3호
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• pp.37.1-37.14
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• 2023

Background: Toll-like receptor (TLR) agonists have been used as adjuvants to modulate immune responses in both animals and humans. Objectives: The objective of this study was to evaluate the combined effects of the TLR 4 agonist monophosphoryl lipid A (MPL) and the TLR 3 agonist polyinosinic:polycytidylic acid (Poly I:C) on equine peripheral blood mononuclear cells (PBMCs), monocyte-derived dendritic cells (MoDCs), and bone marrow-derived mesenchymal stromal cells (BM-MSCs). Methods: The PBMCs, MoDCs, and BM-MSCs collected from three mixed breed horses were treated with MPL, Poly I:C, and their combination. The mRNA expression of interferon gamma (IFN-γ), interleukin (IL)-1β, IL-4, IL-6, IL-8, IL-12p40, tumor necrosis factor alpha (TNF-α), vascular endothelial growth factor (VEGF), and monocyte chemoattractant protein-1 (MCP-1) was determined using real-time polymerase chain reaction. Results: The combination of MPL and Poly I:C significantly upregulated immunomodulatory responses in equine cells/ without cytotoxicity. The combination induced greater mRNA expression of pro-inflammatory cytokines IFN-γ and IL-6 than MPL or Poly I:C stimulation alone in PBMCs. In addition, the combination induced significantly higher mRNA expression of IL-1β, IL-6, and IL-12p40 in MoDCs, and IL-8, MCP-1, and VEGF in BM-MSCs compared to stimulation with a single TLR agonist. Conclusions: The combination of MPL and Poly I:C can be used as a potential adjuvant candidate for vaccines to aid in preventing infectious diseases in horses.

• Journal of Veterinary Science
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• 제24권5호
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• pp.55.1-55.12
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• 2023

Background: Peste des petits ruminants (PPR), caused by the PPR virus (PPRV), is an acute and fatal contagious disease that mainly infects goats, sheep, and other artiodactyls. Peripheral blood mononuclear cells (PBMCs) are considered the primary innate immune cells. Objectives: PBMCs derived from goats were infected with PPRV and analyzed to detect the relationship between PPRV replication and apoptosis or the inflammatory response. Methods: Quantitative real-time polymerase chain reaction was used to identify PPRV replication and cytokines expression. Flow cytometry was conducted to detect apoptosis and the differentiation of CD4⁺ and CD8⁺ T cells after PPRV infection. Results: PPRV stimulated the differentiation of CD4⁺ and CD8⁺ T cells. In addition, PPRV induced apoptosis in goat PBMCs. Furthermore, apoptosis and the inflammatory response induced by PPRV could be suppressed by Z-VAD-FMK and Z-YVAD-FMK, respectively. Moreover, the virus titer of PPRV was attenuated by inhibiting caspase-1-dependent apoptosis and inflammation. Conclusions: This study showed that apoptosis and the inflammatory response play an essential role in PPR viral replication in vitro, providing a new mechanism related to the cell host response.

• Tuberculosis and Respiratory Diseases
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• 제53권1호
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• pp.5-16
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• 2002

연구배경 : 말초혈액 단핵구와 폐포 대식세포가 림프구에 의하여 활성화된 후 탐식된 세포내 결핵균을 제거하는 최종 효과기로 작용하지만 결핵균은 이러한 세포 내에서조차 살아남아 증식한다. 본 연구에서는 인체 감염의 경우와 유사하도록 낮은 감염률(결핵균:포식세포 1:10)을 이용하여 MAC(Mycobacterium avium)과 Mycobaterium tuberculosis $H_{37}Ra$에 대한 단핵구와 폐포 대식세포의 림프구 의존적 결핵균 살해능의 정도를 측정하고자 하였다. 방 법 : 정상인과 폐결핵 환자의 말초혈액 단핵구를 Ficoll gradient 방법을 이용하여 분리하고, 정상인의 폐포 대식세포를 기판지폐포 세척액에서 분리 하여 96well plate에 $1{\times}10^5$ 씩 분주하고 결핵균주 MAC과 $H_{37}Ra$에 낮은 감염률(결핵균:단핵구 1:10)로 감염시켰다. 일부는 비부착세포(림프구)를 림프구:포식세포 10:1 비율로 첨가하여 $37^{\circ}C$, 5% $CO_2$ 배양기에 배양하였다. 1일, 4일, 및 7일째 각각 수확하여 middlebrook 7H10/OADC agar plate에 접종 후 집락이 형성될 때까지 $37^{\circ}C$, 5% $CO_2$ 배양기에 배양하였다. 결핵균의 수는 lysate의 $m{\ell}$(세포수 $10^6$)당 CFU로 표시하였고, 결핵균의 살해능은 log killing ratio[logKR=$log_{10}$(Final CFU/Initial CFU)]로 표시하였다. 배양 1일째 상층액에서 TNF-${\alpha}$, 배양 4일째 상층액에서 ${\gamma}$-IFN을 Elisa kit를 사용하여 측정하였다. 결 과 : 단핵구의 세포내 결핵균 살해능은 정상대조군에 비하여 결핵환자군에서 ${\Delta}logKR$가 MAC -0.4, $H_{37}Ra$ -0.2 정도로 유의하게 증가되어 있었다(p<0.05). 정상대조군에서 폐포 대식세포의 결핵균 살해능은 단핵구에 비하여 ${\Delta}$logKR가 MAC과 $H_{37}Ra$ 모두 -0.2정도로 증가된 경향을 보였다. 단핵구와 폐포 대식세포내 결핵균 살해능은 림프구 의존적으로 림프구 첨가시의 ${\Delta}logKR$는 MAC -0.4, $H_{37}Ra$ -0.5 정도로 포식세포 단독에 비하여 유의하게 증가되었다 (p<0.05). ${\gamma}$-IFN은 말초 단핵구와 폐포 대식세포 단독배양시 분비량이 경미하였으나 림프구 첨가시에는 유의한 분비의 증가를 보였다. 결 론 : 본 연구는 낮은 결핵균 감염률을 이용한 인체 포식세포의 세포내 결핵균 감염 모델이다. 결핵균 살해능은 림프구 의존적이며, 포식세포의 결핵균 탐식 후 7일까지의 배양에서 결핵환자의 단핵구와 PPD 양성 정상인의 폐포 대식세포에 림프구를 첨가한 경우에만 실질적인 균 증식의 제한을 볼 수 있었다.

• Clinical and Experimental Pediatrics
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• 제45권2호
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• pp.247-255
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• 2002

목 적 : 태아 간세포는 면역학적으로 미숙하여 이식에 따른 거부반응이 잘 일어나지 않으므로 조혈모세포 이식, 특히 자궁내 이식의 좋은 공여세포로 이용할 수 있으나 아직 국내에서는 이를 위한 기초연구가 미미한 실정이다. 연구자는 태아 간 단핵세포를 이용하여 면역조직학적 염색과 체외배양 결과를 분석하여 태아 간의 거핵구 집락형성과 관련된 기초자료를 마련하고자 이 연구를 시행하였다. 방 법: 임신중절시 수집한 태아 간조직과 특발성 혈소판 감소성 자반증 및 범혈구 감소증 환아의 골수에서 단핵세포를 분리한 후 $MegaCult^{TM}-C$ 배지에서 12일간 배양 후 성장인자 첨가에 따른 거핵구 집락형성을 관찰하였고, 5일간의 Flask 배양 후 유세포 분석기를 이용하여 CD34, CD41 양성세포를 측정하였으며, methylcellulose 배지를 이용하여 $37^{\circ}C$ 5% $CO_2$ 배양기에서 배양한 후 제 7일과 14일에 성장인자 첨가에 따른 CFU-GM 집락 수를 조사하였다. 결 과 : 태아 간의 단핵세포 수는 재태주령 11주에 비해서 19주에서 더 많았으며 세포생존율은 $91.2{\pm}3.4%$였다. $MegaCult^{TM}-C$ kit를 이용하여 12일간 배양시 재태주령 11주의 태아 간이 19주에 비해서 거핵구 집락 수가 많았고, 4례 모두 성장인자의 첨가에 따른 집락증폭의 상승효과는 없었으며 배양 후 형성된 거핵구 집락 수는 태아 간, 특발성 혈소판 감소성 자반증 환아의 골수, 범혈구 감소증 환아의 골수 순이었다. 거핵구 집락 중 순수집락 수는 태아 간이 특발성 혈소판 감소성 자반증 및 범혈구 감소증 환아의 골수에 비해서 유의하게 높았으며, 순수집락 중 large CFU-Mk의 비율은 태아 간이 특발성 혈소판 감소성 자반증 및 범혈구 감소증 환아의 골수에 비해서 높았으나 유의성은 보이지 않았다. 태아 간 단핵세포를 5일간 flask 배양한 후 CD34 양성세포의 발현율은 증가하였으나 성장인자의 첨가에 따른 집락증폭의 상승효과는 TPO 투여군에서만 있었고, CD41 양성세포의 발현율 역시 배양 후 증가하였으나 성장인자 첨가군에 비해서 대조군의 발현율이 너무 높은 결과를 보였다. Methylcellulose 배지를 이용한 단핵세포 배양에서 CFU-GM 집락 수는 태아 간의 경우 배양 7일에 비하여 14일에 감소하였고, 성장인자의 첨가에 따른 집락증폭의 상승효과는 태아 간에서는 모든 군에서 보였으나 특발성 혈소판 감소성 자반증에서는 GM-CSF 투여군에서만 있었으며 범혈구 감소증 환아의 골수에서는 없었다. 결 론: $MegaCult^{TM}-C$ 배지를 이용하여 태아 간세포를 배양하고 면역조직학적 염색으로 거핵구 집락형성을 성공적으로 관찰하였으며, 태아 간에서 얻은 거핵구 집락이 특발성 혈소판 감소성 자반증 및 범혈구 감소증 환아의 골수에서 얻은 집락에 비하여 집락수나 순수집락의 분포, 집락의 크기 등에서 우수한 것으로 나타났으나 태아 간 조혈모세포 이식의 실제적인 임상적용을 위해서는 체계적인 더 많은 연구가 필요할 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제9권1호
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• pp.106-112
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• 1999

The therapeutic potential of phosphodiesterase 4(PDE4) inhibitors in inflammatory diseases including some autoimmune diseases has been explored recently with some hopeful results. These PDE4 inhibitors are thought to show their anti-inflammatory effect by down-regulating tumor necrosis factor-a (TNF-$\alpha$) production in lymphocytes and macrophages. A high concentration of TNF-$\alpha$has been found in rheumatoid arthritis (RA) synovium and reducing TNF-$\alpha$using biological agents was proven to be an effective RA treatment. To test the possibility of using PDE4 inhibitors for RA treatment, the effects of a newly synthesized PDE4 inhibitor, DWP205505, on TNF-$\alpha$ and IL-10 production was tested in cells isolated from normal peripheral blood and rheumatoid arthritis synovial fluid. Cytokine production was assayed at the protein level by sandwich enzyme-linked immunosorbent assay (ELISA) and at the mRNA expression level by semi-quantitative RT-PCR. Another PDE4 inhibitor, RP73401, was used for comparison. DWP205505 and RP73401 had no harmful effect on cell viability up to 10 $\mu$M concentration during the 24 h culture period. DWP205505 as well as RP73401 significantly reduced TNF-$\alpha$ secretion from lipopolysaccharide (LPS)-stimulated peripheral blood mononuclear cells (pBMC) and synovial fluid mononuclear cells (SFMC). The effect of DWP205505 or RP73401 treatment on the mRNA expression of TNF-$\alpha$ was also studied in LPS-stimulated PBMC and SFMC. TNF-$\alpha$ mRNA expression was increased by LPS stimulation and both of the PDE4 inhibitors suppressed TNF-$\alpha$ mRNA expression. For interleukin-l0 (IL-l0), a little different results were obtained from PBMC and SFMC; IL-l0 secretion was unaffected by LPS stimulation and only minimally affected by both of the PDE4 inhibitors in PBMC. In unstimulated SFMC, DWP205505 and RP73401 slightly enhanced IL-10 secretion, while they reduced IL-l0 secretion from LPS-stimulated SFMC where IL-l0 secretion was a lot higher than unstimulated SFMC. These results suggest that the newly synthesized PDE4 inhibitor DWP205505 may have anti-rheumatoid arthritis activity.

• 대한치과교정학회지
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• 제17권2호
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• pp.223-248
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• 1987

The early tissue reactions in the periodontal tissues of the tension zones following the application of force (30gm) to the maxillary first molar teeth of the albino rats were studied by the light microscopy and electron microscopy The increase of periodontal fibroblasts was evident, particularly in 1 day survival period. Osteoblast differentiation and new bone formation on the alveolar bone surface were occurred from 1 day survival period. Mononuclear phagocytes occurred consistently and in relatively high number adjacent to and at some distance from blood vessel Extensive breakdown of collagen fibers was observed. The increase of the phagocytosis of collagen by the active fibroblasts was evident Also, collagen fibrils were sparse or lost near the macrophage.

• 대한화학회지
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• 제29권5호
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• pp.482-489
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• 1985

오클로로옥소몰리브덴(V)산의 치환퀴놀린윰염을 합성하고 착물의 원소분석, 적외선흡수 스펙트럼, 전자스펙트럼, 전기전도도를 측정하였다. 착물은 2:1전해질이고 치환퀴놀린류는 양이온 상태로 존재하고 있으며 착물의 음이온은 단위체이다. 착물의 결정장전이와 전하이동전이를 해석하였다.

• 대한수의학회지
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• 제5권1호
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• pp.21-25
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• 1965

Throughout the studies the followings were obtained and summarized here. 1. Thickenings of plerua were due to the organized fibrin deposition on the pleural surface. 2. Thickenings of plerura were accompanied with increaseness of fibrous connective tissues in the interlobular and alveolar septa. 3. Eosinophilic inclusions seemed to LCL bodies were observed in the cytoplasms of epithelial cells of bronchi, bronchial glands and in the cytoplasms of mononuclear cells. 4. Histo-pathologically, there were some evidences influenced by viruses and the organisms of Psittacosis-Lymphogranuloma Venereum Group.