규칙 설명 및 명시적 피드백이 구조화된 입력 연습을 통한 한국 학생들의 문법학습에 미치는 효과

Thesis(masters) --서울대학교 대학원 :외국어교육과(영어전공),2010.2.Maste

효율적인 (E)-Alkene 및 Ketovinyl Dipeptide Isostere의 합성

Maste

Mycobacterium tuberculosis 30kDa antigen induces NF-kB activation and cytokine production by

의과학과/석사[한글]결핵균은 세포 내 기생세균으로 숙주의 대식세포에 의해서 파괴되지 않고 만성감염을 유발한다. 즉 흡입에 의해서 체내로 들어가 폐에서 대식세포에 탐식된 후 생존, 증식하여서 결국 폐에 감염원을 생성하기 때문에 대식세포의 반응을 연구하는 것이 결핵균에 대한 면역반응을 이해하는데 몹시 중요하다. 대식세포는 선천면역반응에 우선 관여하는데 대식세포의 수용체 중 Toll-like receptor(TLR)는 미생물에서 존재하는 PAMP(pattern-recognition molecular pattern)를 인지하여 병원체와 자신을 구별한다. TLR ligand 중 특히 그람 음성균 박테리아의 세포벽 성분의 lipopolysaccharide(LPS)와 그람 양성균의 lipoteichoic acid(LTA)가 대표적인 PAMP로서 각각 TLR2와 TLR4와 반응한다. 결핵균 항원도 TLR을 통하여 면역세포를 활성화시킨다는 보고가 있지만 현재까지 TLR과 연관된 연구는 결핵균 다당류 항원 (예; lipoarabinomannan)으로 제한되어 있었다. 그러므로 본 연구에서는 결핵균 단백 항원 중에서 TLR을 자극하는 항원이 있는지를 사람 대식세포주와 TLR2/4 transfectant cell에서 관찰하였다.본 실험에서는 총 9종류의 결핵균 단백항원을 사용하였다. 10, 22, 30, 38 kDa 항원은 정제된 항원이며, 6, 16, 19, 38, Ag85A는 재조합항원이었다. 사람 대식세포주인 THP-1 세포를 대상으로 TLR분자의 발현과 cytokine 생성의 변화를 확인하였을 때, 재조합항원에서는 19kDa 을 제외한 6, 16, 38, Ag85A 항원은 cytokine 생성을 강하게 유도하였다. 또한 정제항원 중에서는 30kDa 항원이 대식세포에서 TNF-α와 IL-6의 생성을 가장 강하게 유도하였다. 이상 9종류의 항원 중 단백 생성 과정에서 포함된 LPS에 의한 영향을 배제하기 위하여 정제항원만을 중심으로 이후 연구를 진행하였다.MAP kinase 활성화를 관찰한 결과 정제 30kDa항원은 ERK의 활성화를 강하게 유도하였고, 정제 30kDa 항원이 대식세포를 자극하는 기전을 파악하기 위하여 TLR2 또는 TLR4 transfectant HEK293 세포를 대상으로 NF-κB 활성화를 측정하였다. 그 결과 정제 30kDa 항원 자극에 의하여 TLR2 transfectant cell에서만 NF-κB 활성화가 강하게 관찰되었고 TLR4 transfectant cell에서는 관찰되지 않았다. TLR2 transfectant cell에서 유도된 NF-κB 활성화는 항-TLR2 항체에 의하여 억제되었다.이상의 결과를 통하여 결핵균 30kDa 항원이 TLR2를 자극함으로써 대식세포의 활성화를 유도하여 IL-6 및 TNF-α를 생성시키는 것으로 추정되었다. 본 연구에서는 결핵균 단백항원 중에서도 TLR2와 반응하는 분자가 있음을 밝혔으며 이는 30kDa 항원이 결핵균 감염 시 인체의 면역반응을 조절할 가능성을 제시한다. [영문]Mycobacterium tuberculosis, the causative agent of tuberculosis, is an intracellular bacterium that is capable of surviving and persisting within host mononuclear cells. Macrophage phagocytosis of M. tuberculosis bacilli is accompanied by activation of the transcription factor NF-κB, secretion of inflammatory mediators, release of the reactive nitrogen intermediate NO, and secretion of several chemokines. Mammalian Toll-like receptor (TLR) proteins have recently been shown to play important roles in host cell response to bacteria and bacterial products in vitro. Mycobacterial products released as PIM, LM, LAM, lipoproteins and other mycobacterial factors may contribute to continued macrophage and dendritic cell(DC) activation through PRR such as TLRs and others. Several published reports have shown that the TLR2 and TLR4 proteins mediate cellular activation by a variety of bacterial products, including gram-negative bacterial lipopolysaccharide(LPS), the mycobacterial glycolipid lipoarabinomannan(LAM), bacteraial lipoproteins, peptidoglycan and zymosan. Up to the present, several mycobacterial antigens can achieve cellular activation through interaction with TLR2.In this study, we used several mycobacterial antigens(purified 10, 22, 30, 38kDa, recombinant 6, 16, 19, 38kDa and Ag85A antigen). The characteristics of the human macrophage cell line THP-1 was investigated solely by stimulation with purified antigen in order to prevent LPS contamination of recombinant antigen in the process of protein synthesis. Purified mycobacterial antigen-induced TLR expression and cytokine production in human PMA-differentiated THP-1 macrophage cells, even without treatment with mycobacterial antigens, increased levels of TLR2 and TLR4 expression. We obtained evidence demonstrating that only 30 kDa antigen induces production of IL-6 or TNF-alpha in THP-1 macrophages cells.We examined the influence of mycobacterial purified antigens on the MAP kinase signaling pathway in THP-1 cells and HEK293-TLR2/4 transfectant cells. When THP-1 cells were treated with 30kDa antigen, strong phosphorylation of ERK occurred. We also found that the 30kDa antigen activated NF-kB in TLR2-transfected cells, not TLR4-transfected cells. Antibody blocking experiments revealed that the inhibition of anti-TLR2 to NF-κB activation in TLR2-transfactant cells.These results indicate that TLR2 may act as an important receptor when stimulated with 30kDa antigen in macrophage infection.ope

Risk factors of septic shock in chemotherapy induced febrile neutropenia patients who visited an emergency department

목적: 항암화학요법 후 열성 호중구감소증으로 응급실에 내원한 암환자의 특성을 분석하고 패혈성쇼크 발생 위험요인을 확인하고자 시행된 후향적 조사연구이다. 대상 및 방법: 2018 년 1 월 1 일부터 2018 년 12 월 31 일까지 항암화학요법 후 열성 호중구감소증으로 서울 소재 일 상급 종합 병원 응급실에 내원한 430 건의 암환자 의무기록을 조사하였다. 대상자의 특성은 증례보고서를 이용하여 전자의무기록으로 조사하였다. 대상자의 일반적 특성, 항암화학요법 관련 특성, 임상적 특성, 응급실 관련 특성을 조사한 후 대상자의 패혈성쇼크 비발생군과 발생군의 특성 차이를 확인하고, 패혈성쇼크 위험요인을 파악하였다. 수집된 자료는 SPSS 25.0 으로 분석하였으며 기술통계, t-test, Mann-Whitney U test, Chi-Square test, Fisher’s exact test, logistic regression 을 이용하였다. 결과: 총 430 건의 열성 호중구감소증 사례 중 패혈성쇼크 비발생군은 388 건(90.2%), 발생군은 42 건(9.8%)이었다. 항암화학요법 후 열성 호중구감소증으로 응급실에 내원한 암환자의 패혈성쇼크 위험요인은 남성(Odds Ratio [OR]: 2.272, 95% Confidence Interval [CI]: 1.153~4.479, p=.018), Eastern Cooperative Oncology Group Performance Status 2, 3, 4 등급(OR: 3.553, CI: 1.748~7.221, p<.001), 마지막 항암화학요법을 받은 후 응급실 내원까지 경과일(OR: 0.923, CI: 0.853~0.999, p=.047), 수축기 혈압(OR: 0.960, CI: 0.932~0.988, p=.006), 균혈증(OR: 21.602, CI: 7.204~64.773, p<.001), KTAS 1, 2 단계(OR: 18.663, CI: 6.222~55.976, p<.001)이었다. 결론: 본 연구를 통해 항암화학요법 후 열성 호중구감소증으로 응급실에 내원한 암환자의 특성과 패혈성쇼크 위험요인을 확인할 수 있었다. 임상에서 이러한 특성을 고려하여 환자를 사정하고 패혈성쇼크 위험을 최소화하기 위해 노력해야 할 것이다.Maste

Bone tissue engineering using human fetal cartilage-derived progenitor cells and porcine cartilage acellular matrix scaffold

학위논문(석사)--아주대학교 일반대학원 :분자과학기술학과,2020. 81. Introduction 1 2. Materials and Methods 3 2.1 Cell isolation and culture 3 2.2 Fabrication and characterization of porcine cartilage-derived acellular matrix (CAM) powder scaffolds 4 2.3 Seeding hFCPCs on a CAM scaffold 4 2.4 Osteogenic differentiation of hFCPCs on a CAM scaffold 5 2.5 Alkaline Phosphatase (ALP) Activity 5 2.6 Quantitative Real Time Polymer (qRT-PCR) analysis 6 2.7 Histological evaluation 6 2.8 Immunohistochemistry analysis 7 2.9 The effect on bone regeneration by hFCPCs-seeded CAM scaffold in rabbit tibia; gross view and micro-CT analysis 7 2.10 Histological evaluation 9 2.11 Immunohistochemistry analysis 9 2.12 Statistical analysis 10 3. Results 14 3.1 Fabrication of porous CAM scaffolds 14 3.2 preparation of cell-seeded CAM scaffolds 17 3.3 In vitro osteogenesis of hFCPCs in CAM scaffolds 17 3.4 In vivo bone repair using construct fabricated with hFCPCs-seeded CAM scaffolds on a rabbit tibial defect 21 3.5 Evaluation of new bone formation for hFCPCs-seeded CAM scaffolds on tibia defect in rabbit model using micro-CT 23 3.6 hFCPCs-seeded CAM scaffold promoted new bone formation in rabbits 25 4. Discussion 29 References 32MasterPURPOSE. In this study, engineered artificial bone tissue was prepared by using human fetal cartilage-derived progenitor cells (hFCPCs) and scaffold made of extracellular matrix derived from porcine cartilage. In many studies, it has been known that markers related to osteogenic differentiation are up-regulated when inoculating mesenchymal stem cells into the extracellular matrix scaffold and inducing osteogenic differentiation in vitro. It also reported that mesenchymal stem cells having multi-lineage differentiative potency were inoculated into cartilage-derived extracellular matrix scaffolds to fabricate artificial bone tissue in vivo. With this regard, this study aimed to confirm the possibility that artificial bone tissue can be produced by inoculating hFCPCs with osteogenic differentiation ability into extracellular matrix scaffolds derived from porcine cartilage. METHODS. Porous cartilage extracellular matrix scaffolds were prepared through a salt-extraction method. In vitro environment, hFCPCs and hBMSCs were inoculated in a porcine cartilage extracellular matrix scaffold and in vitro cultured in a bone differentiation medium to induce bone differentiation. In addition, in order to see the bone formation in vivo of the artificial bone tissue thus produced, a defect was observed in the tibia of a rabbit after transplantation. RESULTS. As a result of in vitro experiments, in the same environment, the hFCPCs group showed no significant difference in the ALP activity and bone formation gene expression of the hBMSCs group at 0, 7, 14 days after differentiation. On day 21, the expression of the Osterix and Osteocalcin genes was significantly higher in hFCPCs than in hBMSCs. In vivo experiments, hFCPCs were used to confirm that the porcine cartilage-derived extracellular matrix scaffolds inoculated with hFCPCs helped bone regeneration when transplanted into a tibial defect in a rabbit and were compared with the group treated with no treatment as a control group. As a result of analysis through micro-computed tomography and histological staining, the hFCPCs group showed a tendency to increase rapidly regenerated bone volume when going from 8 to 12 weeks. And at 12 weeks, it was shown that the hFCPCs group induced significantly more bone regeneration than rBMSCs. And also, the group transplanted with hFCPCs after 12 weeks showed an effect on bone regeneration similar to the negative control without any treatment. Through histological staining, newly formed bone structures were identified at artificially created defect sites, and the fact that the transplanted hFCPCs differentiated into osteoblasts and angiogenesis in tissues was also confirmed by the method of immunostaining. CONCLUSIONS. The hFCPCs-seeded CAM scaffold group showed relatively low regenerated bone volume at 4 and 8 weeks when compared to the negative control with no treatment. At 12 weeks, the hFCPCs group did not show any significant difference from the negative control group. In conclusion, when the bone tissue was prepared by inoculating hFCPCs in a cartilage-derived extracellular matrix scaffold of pigs, the transplantation into a tibial defect of a rabbit did not prove significantly better bone tissue regeneration than a negative control group, the conclusion was not reached. Based on these results, it is judged that positive results will be obtained for the evaluation of bone regeneration ability of the experimental group by supplementing the experimental design in the future