류마티스 관절염 환자의 말초혈액 단핵세포에서 Phosphoinositide 3-Kinase (PI3K)/Akt와 Nuclear Factor KappaB (NF-κB) 신호전달을 통한 IL-17 생성조절

김경운,조미라,이상헌,민소연,박미경,박성환,주대명,김호연,Kim, Kyoung-Woon,Cho, Mi-La,Lee, Sang-Heon,Min, So-Youn,Park, Mi Kyung,Park, Sung-Hwan,Jue, Dae-Myung,Kim, Ho-Youn 대한면역학회 2003 Immune Network Vol.3 No.4

Inflammatory mediators has been recognized as an important role in the pathogenesis of rheumatoid arthritis (RA). IL-17 is increasingly recognized as an important regulator of immune and inflammatory responses, including induction of proinflammatory cytokines and osteoclastic bone resorption. Evidence of the expression and proinflammatory activity of IL-17 has been demonstrated in RA synovium and in animal models of RA. However, the signaling pathways that regulate IL-17 production remain unknown. In the present study, we investigated the role of the phosphatidylinositol 3 kinase (PI3K)-Akt pathway in the regulation of IL-17 production in RA. PBMC were separated from RA (n=24) patients, and stimulated with various agents (anti CD3, anti CD28, PHA, ConA, IL-15). IL-17 levels were determined by sandwich ELISA and RT-PCR. The production of IL-17 was significantly increased in cells treated with anti-CD3 antibody, PHA, IL-15 or MCP-1 (P<0.05). ConA also strongly induced IL-17 production (P<0.001), whereas TNF-alpha, IL-1beta, IL-18 or TGF-beta did not. IL-17 was detected in the PBMC of patients with osteoarthritis (OA) but their expression levels were much lower than those of RA PBMC. Anti-CD3 antibody activated the PI3K-Akt pathway and activation of the PI3K-Akt pathway resulted in a pronounced augmentation of nuclear factor kappaB ($NF-{\kappa}B$). IL-17 production by activated PBMC in RA is completely or partially blocked in the presence of $NF-{\kappa}B$ inhibitor PDTC and PI3K-Akt inhibitor, wortmannin and LY294002, respectively. Whereas the inhibition of AP-1 and extracellular signal-regulated kinase (ERK)1/2 did not affect IL-17 production. These results provide new insight into that PI3K/Akt and $NF-{\kappa}B$ dependent signal transduction pathway could be involved in the overproduction of key inflammatory cytokine, IL-17 in rheumatoid arthritis.

봉약침과 홍화약침을 이용한 원형탈모증(圓形脫毛症) 치험 1예

김경운,이경민,임성철,정태영,한상원,서정철,Kim, Kyung-Woon,Lee, Kyung-Min,Lim, Seong-Chul,Jung, Tae-Young,Han, Sang-Won,Seo, Jung-Chul 대한약침학회 2004 Journal of pharmacopuncture Vol.7 No.2

Objective : Alopecia areata is a common disease and the difficulty of its medical management is well known. This study was designed to investigate the effect of bee venom and Carthami Flos herbal acupuncture. Method : The patient was managed by bee venom, carthami-flos herbal acupuncture and General acupunture. Herbal acupuncture was injected subcutaneously into the lesion. We checked involvement in plaque, according to evaluation of the effect on alopecia areata. Results : After 5 month of treatment, the lesion had been replaced with new terminal hair. Evaluation of the effect on alopecia areata The score changed from 0 to 3 point. Conclusion : These result suggest that bee venom and Carthami Flos herbal acupuncture has good effect on alopecia areata. But further studies are required to concretely prove the effectiveness of bee venom and Carthmi Flos herbal acupuncture for treating alopecia areata.

(주) 블루플래닛

김경운(Kyung Woon Kim) 한국자동차공학회 2008 오토저널 Vol.30 No.5

기체 블로우 다운의 동적 모델링 및 분석

김경운 ( Kyung Woon Kim ),서지원 ( Ji Won Seo ),황성원 ( Sung Won Hwang ),이윤주 ( Yun Ju Lee ),문영식 ( Young Sik Moon ) 한국화학공학회 2016 Korean Chemical Engineering Research(HWAHAK KONGHA Vol.54 No.3

본 연구는 vessel blowdown 시 발생하는 온도와 압력 변화를 보다 정확히 예측하기 위하여 기존에 개발된 dynamicmodel을 기반으로 새로운 모델을 개발하고, 개발한 모델의 정확도를 높이기 위하여 vessel 내부의 흐름이 층류일 때와 난류일 때를 모두 고려하여 vessel 벽면으로부터 기체로의 열 전달량을 계산하였다. 효율적인 열역학 계산을 위해 일체의 계산식은 압력이 감소하는 단계 별로 나누어 진행하였으며 계산의 부담을 덜어주면서 계산의 정확도를 유지하기 위한 압력변화 size를 결정하였다. 개발한 모델에 Peng-Robinson equation과 Soave-Redlich-Kwong equation을 적용하여 각각의 경우에 따른 결과의 차이를 비교하였다. 마지막으로 모델의 검증을 위해 Haque et al.의 실험조건을 동일하게 적용하여 실험 결과와 시뮬레이션 결과를 비교 하였으며, 이를 통해 모델의 정확도를 입증하였다. For accurate estimation over the change of pressure and temperature of the vessel during blowdown period, a new dynamic blowdown model was developed in this work. In particular, heat transfer from the vessel wall to discharge gas at both laminar or turbulent flow in the vessel was embedded to the model to increase the accuracy of blowdown estimation. For thermodynamics, the whole blowdown period was discretized into finite pressure decrement steps, and the step size was adjusted so that the calculation can be more efficiently carried out, while maintaining the model``s accuracy. Both Peng-Robinson and Soave-Redlich-Kwong equation of states were applied to the model, and the results were compared each other. Finally, the simulation results was compared with Haque and coworkers`` experimental results, and it proved high accuracy of the model.

A Premier Global Specialty Chemical & Materials 한국그레이스(주)

서경화(Kyung-Hwa Suh),김경운(Kyung-Woon Kim) 한국콘크리트학회 2009 콘크리트학회지 Vol.21 No.1

Identification of the Pig β-1,3-N-acetylglucosaminyltransferase 1 (pB3GNT1) that is Involved in Poly-N-acetyllactosamine (poly-LacNAc) Synthesis

Ji-Youn Kim(김지윤),Hwan-Jin Hwang(황환진),Hak-Jae Chung(정학재),Shinichi Hochi(신이치호치),Mi-Ryung Park(박미령),Sung June Byun(변승준),Keon Bong Oh(오건봉),Hyeon-Yang(양 현),Kyung-Woon Kim(김경운) 한국생명과학회 2018 생명과학회지 Vol.28 No.4

당 단백질에 붙어 있는 당사슬 구조는 형질전환 돼지 유즙으로 분비되는 의약용 단백질의 생물학적 활성, 안정성 그리고 안전성에 영향을 줄 수 있다. 형질전환 동물을 이용한 치료용 당 단백질 생산은 유선 세포에서 이루어지는 당사슬 부가능력에 의해 제한되며, 균일한 당사슬 형태를 가지는 당 단백질 생산은 도전 과제로 남아있다. β-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) 유전자는 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 단백질 당화기작에 중요한 효소이지만, 돼지 당 전이효소에 대한 정보는 매우 제한적이다. 따라서, 돼지 B3GNT1 (pB3GNT1) 유전자를 클로닝하고 N-아세틸글루코사민에 갈락토오스 잔기를 부착시키는 기능적 특성을 조사하였다. 몇가지 다른 프라이머를 사용하여 전체 전사영역(ORF)을 함유하는 부분적인 pB3GNT1 mRNA 염기서열을 간 조직으로부터 분리하였다. 클로닝 된 pB3GNT1의 ORF는 1,248개의 뉴클레오티드를 가지며, 415개 아미노산 잔기로 구성되어 있었다. pB3GNT1 유전자의 장기별 발현특성은 성돈 및 자돈의 여러 기관에서 분석하였다. pB3GNT1 mRNA 발현 수준은 심장, 소장 보다는 근육에서 높았지만 폐에서는 낮았다. pB3GNT1의 기능적 특성 분석을 위해 돼지 신장 세포주(PK-15)에서 pB3GNT1 유전자의 안정적인 발현을 확립하였다. 그 결과, PK-15 세포에서 pB3GNT1 발현에 의한 당화 패턴은 총 시알산 증가에는 영향을 미치지 않지만, poly-N-아세틸글루코사민은 증가하는 것으로 나타났다. 본 연구는 생물반응기로 형질전환 돼지를 이용할 때 희망하는 당사슬을 부가하여 치료 가능성을 높이며 개선된 활성을 나타내는 당단백질 생산에 도움이 될 것이다. The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The β-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

류마티스관절염 활막세포에서 NF-${\kappa}B$ 신호전달을 통한 MIF의 SDF-1 생성 유도

조미라,박미경,김경운,오혜좌,이선영,박진실,허유정,주지현,민준기,이상헌,박성환,김호연,Cho, Mi-La,Park, Mi-Kyung,Kim, Kyoung-Woon,Oh, Hye-Jwa,Lee, Seon-Yeong,Park, Jin-Sil,Heo, Yu-Jung,Ju, Ji-Hyeon,Min, Jun-Ki,Lee, Sang-Heon,Park, Sung-Hwa 대한면역학회 2007 Immune Network Vol.7 No.1

Background: Stromal cell-derived factor (SDF)-1 is a potent chemoattractant for activated T cells into the inflamed Rheumatoid arthritis (RA) synovium. To determine the effect of macrophage migration inhibitory factor (MIF) on the production of SDF-1 in the inflamed RA synovium. Methods: The expression of SDF-1 and MIF in RA and Osteoarthritis (OA) synovium was examined by immunohistochemical staining. The SDF-1 was quantified by RT-PCR and ELISA after RA fibroblast like synoviocyte (FLS) were treated with MIF in the presence and absence of inhibitors of intracellular signal molecules. The synovial fluid (SF) and serum levels of MIF and SDF-1 in RA, OA and healthy control were measured by ELISA. Results: Expression of SDF-1 and MIF in synovium was higher in RA patients than in OA patients. The production of SDF-1 was enhanced in RA FLS by MIF stimulation. Such effect of MIF was blocked by the inhibitors of NF-${\kappa}B$. Concentrations of SDF-1 in the serum and SF were higher in RA patients than in OA patients and healthy control. SDF-1 and MIF was overexpressed in RA FLS, and MIF could up-regulate the production of SDF-1 in RA FLS via NF-${\kappa}B$-mediated pathways. Conclusion: These results suggest that an inhibition of interaction between MIF from T cells and SDF-1 of FLS may provide a new therapeutic approach in the treatment of RA.

봉약침 요법을 이용한 인대이완 환자 치험 3례 보고

이은경(Eun Kyung Lee),김재수(Jae Soo Kim),김경운(Kyung Woon Kim) 한의병리학회 2007 동의생리병리학회지 Vol.21 No.6

돼지 유래의 β-1,4-N-acetylglucosaminyltransferase A (MGAT4A) 유전자의 동정 및 기능 분석

김지윤(Ji-Youn Kim),황환진(Hwan-Jin Hwang),정학재(Hak-Jae Chung),박미령(Mi-Ryung Park),변승준(Sung June Byun),김경운(Kyung-Woon Kim) 한국생명과학회 2016 생명과학회지 Vol.26 No.3

인체치료용 당단백질은 바이오의약품에 있어서 중요한 요소이며 특히, 단백질 말단에 결합되어있는 시알산은 의약품의 체내 활성이나 안정성에 큰 영향을 미친다. 최근 돼지유즙은 바이오의약품을 생산하기 위한 생체반응기로써 주목받고 있다. β-1,4-N-acetylglucosaminyltransferase A (MGAT4A)는 재조합단백질이 가지는 시알산 함량을 늘리기 위한 필수 효소 중 하나이다. 그러나 돼지 MGAT4A는 아직도 그 서열이나 기능이 불분명하다. 따라서 이번 연구에서는 돼지 MGAT4A의 서열 동정 및 기능분석을 하였다. 돼지 MGAT4A는 535개의 아미노산을 코딩하는 1,638개의 염기서열로 이루어져 있으며, 일반적인 당전이효소들의 특징인 type II membrane topology의 특성을 가지고 있다. Real-time PCR분석법을 통하여, 지금까지 알려진 것과는 조금 다르게 MGAT4A 유전자가 간이나 유선에서 많은 발현을 보였으나 소장이나 위, 방광에서는 그 발현량이 적다는 것도 확인하였다. 또한, 효소의 기능 분석을 위하여 PK-15 세포주에서 MGAT4A효소의 과발현을 유도하였다. 과발현이 확인된 세포주로 렉틴을 이용한 면역형광염색법과 ELISA방법으로 MGAT4A효소의 과발현이 β-1,4-N-acetylglucosamine의 함량을 증가시켰음을 확인하였다. 또한 기질반응을 통해 bi-antennary structure에 대한 반응성도 확인하였으며, MGAT4A의 과발현이 유도된 세포에서 약 3배 이상의 높은 기질 반응성을 보였다. 이러한 연구는 생체반응기로써 돼지를 이용하는데 있어서 중요한 기반이 될 것이라고 생각한다. Glycan modification is important in pharmaceutical industry. Especially, sialic acid affects the bioactivity and stability of medicine. Milk of pig has been used as bioreactor to produce various pharmaceutical proteins. Therefore, it is necessary to modify the glycan chain in pig mammary grand. β-1,4-N-Acetylglucosaminyltransferase A (pMGAT4A) is one of the essential enzymes for increase of sialic acid content, but pig MGAT4A is unclear. In this study, the pMGAT4A was identified and characterized. The pMGAT4A has 1638 nucleotides encoding 535 amino acids and type II membrane topology, which is one of the common features in many glycosyltransferases. The gene was strongly expressed in liver and mammary gland, whereas was weakly expressed in small intestine, stomach and bladder. For functional test, HA-tagged MGAT4A was over-expressed in porcine kidney (PK-15) cell line. Forced expression of pMGAT4A gene was identified by qPCR, and we identified that pMGAT4A is located in Golgi complex by co- staining with HA antibody and BODIPY TR ceramide. In addition, we identified the increase of mannose-β-1,4-N-acetylglucosamine structure by ELISA and immunofluorescence using Datura stramonium agglutinin (DSA), which recognizes mannose-β-1,4-Nacetylglucosamine. Through the specific activity analysis, we showed that pMGAT4A modified bi-antennary to tri-antennary. This event affects sialic acid content. Therefore, we thought that over-expression of pMGAT4A will be necessary in pig mammary grand for improved medicine.

좌섬요통에 대한 BU 약침의 효과

서보명,이윤경,최성훈,김경운,윤종석,임성철,정태영,한상원,서정철,Seo, Bo-Myung,Lee, Yoon-Kyung,Choi, Sung-Hoon,Kim, Kyung-Woon,Yoon, Jong-Seok,Lim, Seong-Chul,Jung, Tae-Young,Han, Sang-Won,Seo, Jung-Chul 대한약침학회 2005 Journal of pharmacopuncture Vol.8 No.2

Objectives : This study was designed to estimate the effects of BU herbal acupuncture theraphy on low back pain of sprain patients by using Visual Analogue Scale(VAS) and pressure algometer. Methods : 13 volunteers were applied to BU herbal acupuncture stimulation on the BL23, BL24, BL25 and BL26 acupoints. The degree of improvement of low back pain of sprain was evaluated by VAS and pain pressure threshold(at the points of BL23 and BL24 before treatment, before 3rd treatment(after herbal acupuncture). Results : In VAS scores, there were significant differences between before and after herbal acupuncture stimulation. But there was no significant difference between before and after herbal acupuncture stimulation in pain threshold. Conclusions : The effectiveness of BU herbal acupuncture theraphy on low back pain of sprain was shown through VAS and pressure algometer. These imply that BU herbal acupuncture theraphy may be useful for low back pain of sprain. Further study is needed about BU herbal acupuncture.