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Leonurus sibiricus L. ethanol extract promotes osteoblast differentiation and inhibits osteoclast formation

  • Jae‑Hyun Kim;Minsun Kim;Hyuk‑Sang Jung;Youngjoo Sohn
    • International Journal of Molecular Medicine
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    • v.44 no.3
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    • pp.913-926
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    • 2019
  • Leonurus sibiricus L. (LS) is a medicinal plant used in East Asia, Europe and the USA. LS is primarily used in the treatment of gynecological diseases, and recent studies have demonstrated that it exerts anti-inflammatory and antioxidant effects. To the best of our knowledge, the present study demonstrated for the first time that LS may promote osteoblast differentiation and suppress osteoclast differentiation in vitro, and that it inhibited lipopolysaccharide (LPS)-induced bone loss in a mouse model. LS was observed to promote the osteoblast differentiation of MC3T3-E1 cells and upregulate the expression of runt-related transcription factor 2 (RUNX2), a key gene involved in osteoblast differentiation. This resulted in the induction of the expression of various osteogenic genes, including alkaline phosphatase (ALP), osteonectin (OSN), osteopontin (OPN), type I collagen (COL1) and bone sialoprotein (BSP). LS was also observed to inhibit osteoclast differentiation and bone resorption. The expression levels of nuclear factor of activated T-cells 1 (NFATc1) and c-Fos were inhibited following LS treatment. NFATc1 and c-Fos are key markers of osteoclast differentiation that inhibit receptor activator of nuclear factor-κB ligand (RANKL)-induced mitogen-activated protein kinase (MAPKs) and nuclear factor (NF)-κB. As a result, LS suppressed the expression of osteoclast-associated genes, such as matrix metallopeptidase-9 (MMP-9), cathepsin K (Ctsk), tartrate-resistant acid phosphatase (TRAP), osteoclast-associated immunoglobulin-like receptor (OSCAR), c-src, c-myc, osteoclast stimulatory transmembrane protein (OC-STAMP) and ATPase H+ transporting V0 subunit d2 (ATP6v0d2). Consistent with the in vitro results, LS inhibited the reduction in bone mineral density and the bone volume/total volume ratio in a mouse model of LPS-induced osteoporosis. These results suggest that LS may be a valuable agent for the treatment of osteoporosis and additional bone metabolic diseases.

Quantum transport of doped rough-edged graphene nanoribbons FET based on TB-NEGF method

  • K.L. Wong;M.W. Chuan;A. Hamzah;S. Rusli;N.E. Alias;S.M. Sultan;C.S. Lim;M.L.P. Tan
    • Advances in nano research
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    • v.17 no.2
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    • pp.137-147
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    • 2024
  • Graphene nanoribbons (GNRs) are considered a promising alternative to graphene for future nanoelectronic applications. However, GNRs-based device modeling is still at an early stage. This research models the electronic properties of n-doped rough-edged 13-armchair graphene nanoribbons (13-AGNRs) and quantum transport properties of n-doped rough-edged 13-armchair graphene nanoribbon field-effect transistors (13-AGNRFETs) at different doping concentrations. Step-up and edge doping are used to incorporate doping within the nanostructure. The numerical real-space nearest-neighbour tight-binding (NNTB) method constructs the Hamiltonian operator matrix, which computes electronic properties, including the sub-band structure and bandgap. Quantum transport properties are subsequently computed using the self-consistent solution of the two-dimensional Poisson and Schrödinger equations within the non-equilibrium Green's function method. The finite difference method solves the Poisson equation, while the successive over-relaxation method speeds up the convergence process. Performance metrics of the device are then computed. The results show that highly doped, rough-edged 13-AGNRs exhibit a lower bandgap. Moreover, n-doped rough-edged 13-AGNRFETs with a channel of higher doping concentration have better gate control and are less affected by leakage current because they demonstrate a higher current ratio and lower off-current. Furthermore, highly n-doped rough-edged 13-AGNRFETs have better channel control and are less affected by the short channel effect due to the lower value of subthreshold swing and drain-induced barrier lowering. The inclusion of dopants enhances the on-current by introducing more charge carriers in the highly n-doped, rough-edged channel. This research highlights the importance of optimizing doping concentrations for enhancing GNRFET-based device performance, making them viable for applications in nanoelectronics.

Nondestructive Evaluation and Microfailure Mechanisms of Single Fibers/Brittle Cement Matrix Composites using Electro-Micromechanical Technique and Acoustic Emission (Electro-Micromechanical 시험법과 Acoustic Emission을 이용한 단섬유/시멘트 복합재료의 미세파괴 메커니즘과 비파괴적 평가)

  • 박종만;이상일;김진원;윤동진
    • Composites Research
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    • v.14 no.3
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    • pp.18-31
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    • 2001
  • Interfacial and microfailure properties of the modified steel, carbon and glass fibers/cement composites were investigated using electro-pullout test under tensile and compressive tests with acoustic emission (AE). The hand-sanded steel composite exhibited higher interfacial shear strength (IFSS) than the untreated and even neoalkoxy zirconate (Zr) treated steel fiber composites. This might be due to the enhanced mechanical interlocking, compared to possible hydrogen or covalent bonds. During curing process, the contact resistivity decreased rapidly at the initial stage and then showed a level-off. Comparing to the untreated case, the contact resistivity of either Zr-treated or hand-sanded steel fiber composites increased to the infinity at latter stage. The number of AE signals of hand-sanded steel fiber composite was much more than those of the untreated and Zr-treated cases due to many interlayer failure signals. AE waveforms for pullout and frictional signals of the hand-sanded composite are larger than those of the untreated case. For dual matrix composite (DMC), AE energy and waveform under compressive loading were much higher and larger than those under tensile loading, due to brittle but well-enduring ceramic nature against compressive stress. Vertical multicrack exhibits fur glass fiber composite under tensile test, whereas buckling failure appeared under compressive loading. Electro-micromechanical technique with AE can be used as an efficient nondestructive (NDT) method to evaluate the interfacial and microfailure mechanisms for conductive fibers/brittle and nontransparent cement composites.

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Analysis of Stromal Cells Developed from Cord Blood CD34+ Cells (제대혈 CD34+ 세포에서 유래된 지지세포의 분석)

  • Ryu, Kyung-Ha;Park, Se-Jin;Kim, Kyung Hyo;Seoh, Ju-Young;Khan, Mohammad;Shin, Hee-Young;Ahn, Hyo-Seop
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.87-94
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    • 2001
  • Background: Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of cord blood (CB) hematopoietic stem cells for transplantation. As well as stem cell number, stromal cells are necessary for functional maturation of hematopoiesis. The purpose of this study was to analyze the development of stromal cells during ex vivo expansion of CB $CD34^+$ cells. Methods : $CD34^+$ cells were purified from CB by magnetic bead selection. The levels of of interleukin-3, interleukin-$1{\beta}$, interleukin-6, granulocyte macrophagecolony stimulating factor and tumor necrosis factor-${\alpha}$ were measured in culture supernatants on 0, 1, 2, and 3 weeks, using ELISA techniques. CB $CD34^+$ cells were expanded in Iscoves modified Dulbeccos medium in the presence of several cytokines. The expression of E-selectin, vascular cell adhesion molecule-1, intercellular adhesion molecule-1, platelet/endothelial cell adhesion molecule-1, von Willebrand factor, vimentin, and CD14 in newly developed stromal cells was examined by immunocytochemical method. Relevant extracellular matrix (ECM) proteins and proper cytokines were also assayed for the most suitable condition for expansion of stromal cells. Results: Several cytokines were found to have been produced by CB $CD34^+$ cells as well as bone marrow-derived $CD34^+$ cells. During ex vivo expansion of CB $CD34^+$ cells, stromal cells appeared in the culture by day 4 and expanded over the following 7-10 days before being confluent by day 2 1. These cells expressed surface markers characteristic of cells of endothelial lineage. Furthermore, these stroaml cells also expanded effectively when treated with thrombopoietin+flt-3 ligand+stem cell factor+leukemia inhibitory factor or 0.1% poly-L-lysine-coated wells. Conclusion: Stromal cells were developed during ex vivo expansion of CB $CD34^+$ cells and that this development could be enhanced further by treating the stromal cells with cytokines or ECM.

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Establishment of a Radiation-Induced Fibrosis Model in BALB/c Mice (BALB/c 마우스를 이용한 방사선섬유증 모델 확립)

  • Ryu, Seung-Hee;Lee, Sang-Wook;Moon, Soo-Young;Oh, Jeong-Yoon;Yang, Youn-Joo;Park, Jin-Hong
    • Radiation Oncology Journal
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    • v.28 no.1
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    • pp.32-38
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    • 2010
  • Purpose: Although radiation-induced fibrosis is one of the common sequelae occurring after irradiation of skin and soft tissues, the treatment methods are not well standardized. This study aimed to establish the skin fibrosis mouse model by fractionated radiation for the further mechanism studies or testing the efficacy of therapeutic candidates. Materials and Methods: The right hind limbs of BALB/c mice received two fractions of 20 Gy using a therapeutic linear accelerator. Early skin damages were scored and tissue fibrosis was assessed by the measurement of a leg extension. Morphological changes were assessed by H&E staining and by Masson's Trichrome staining. TGF-${\beta}1$ expression from soft tissues was also detected by immunohistochemistry and PCR. Results: Two fractions of 20 Gy irradiation were demonstrated as being enough to induce early skin damage effects such as erythema, mild skin dryness, dry and wet desquamation within several weeks of radiation. After 13 weeks of irradiation, the average radiation-induced leg contraction was $11.1{\pm}6.2mm$. Morphologic changes in irradiated skin biopsies exhibited disorganized collagen and extracellular matrix fibers, as well as the accumulation of myofibroblasts compared to the non-irradiated skin. Moreover, TGF-${\beta}1$ expression in tissue was increased by radiation. Conclusion: These results show that two fractions of 20 Gy irradiation can induce skin fibrosis in BALB/c mice accompanied by other common characteristics of skin damages. This animal model can be a useful tool for studying skin fibrosis induced by radiation.

Protection of UV-derived Skin Cell Damage and Anti-irritation Effect of Juniperus chinensis Xylem Extract (향나무추출물의 광손상으로부터 피부세포 보호와 자극완화 효과에 대한 연구)

  • 김진화;박성민;심관섭;이범천;표형배
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.1
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    • pp.63-71
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    • 2004
  • The human skin is constantly exposed to environmental irritants such as ultraviolet, smoke, chemicals. Free radicals and reactive oxygen species (ROS) caused by these environmental facts play critical roles in cellular damage. These irritants are in themselves damaging to the skin structure but they also participate the immensely complex inflammatory reaction. The purpose of this study was to investigate the skin cell protective effect of Juniperus chinensis xylem extract on the UV and SLS-induced skin cell damages. We tested free radical and superoxide scavenging effect in vitro. We found that Juniperus chinensis xylem extracts had potent radical scavenging effect by 98% at 100 $\mu\textrm{g}$/mL. Fluorometric assays of the proteolytic activities of matrix metalloproteinase-l(MMP-1, collagenase) were performed using fluorescent collagen substrates. UV A induced MMP-1 synthesis and activity were analyzed by enzyme-linked immunosorbent assay (ELISA) and gelatin-based zymography in skin fibroblasts. The extract of Juniperus chinensis showed strong inhibitory effect on MMP-1 activities by 97% at 100 $\mu\textrm{g}$/mL and suppressed the UVA induced expression of MMP-1 by 79% at 25 $\mu\textrm{g}$/mL. This extract also showed strong inhibition on MMP-2 activity in UVA irradiated fibroblast by zymography. We also examined anti-inflammatory effects by the determination test of proinflammatory cytokine, interleukin 6 in HaCaT keratinocytes. In this test Juniperus chinensis decreased expression of interleukin 6 about 30%. Expression of prostaglandin E$_2$, (PGE$_2$) after UVB irradiation was measured by competitive enzyme immunoassay (EIA) using PGE$_2$ monoclonal antibody. At the concentrations of 5-50 $\mu\textrm{g}$/mL of the extracts, the production of PGE$_2$ by HaCaT keratinocytes (24 hours after 10 mJ/$\textrm{cm}^2$ UVB irradiation) was significantly inhibited in culture supernatants (p〈0.05). The viability of cultured HaCaT keratinocytes was significantly reduced at the doses of above 10 mJ/$\textrm{cm}^2$ of UVB irradiation, but the presence of these extracts improved cell viability comparing to control after UVB irradiation. We also investigated the protective effect of this extract in sodium lauryl sulfate (SLS)-induced irritant skin reactions from 24 hour exposure. Twice a day application of the extract for reducing local inflammation in human skin was done. Irritant reactions were assessed by various aspects of skin condition, that is, erythema (skin color reflectance) and transepidermal water loss (TEWL). After 5 days the extract was found to reduce SLS-induced skin erythema and improve barrier regeneration when compared to untreated symmetrical test site. In conclusion, our results suggest that Juniperus chinensis can be effectively used for the prevention of UV and SLS-induced adverse skin reactions such as radical production, inflammation and skin cell damage.

The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1276-1289
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    • 2017
  • This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

A Literature Review of Fish Feeding Research in the Coast - Estuary Areas of Korean Peninsula (우리나라 연안-하구에 서식하는 어류의 식성 관련 연구 현황)

  • Jo, Hyunbin;Kim, Dong-Kyun;Park, Kiyun;Lee, Wan-Ok;Kwak, Ihn-Sil
    • Korean Journal of Ecology and Environment
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    • v.52 no.2
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    • pp.126-135
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    • 2019
  • We carried out a literature review of fish food sources research in the coast - estuary areas of Korean peninsula for the last fifty years(1969~2018). A total of 101 research papers were reviewed, of which 81 were published in domestic journals, 7 were puliched in domestic reports and 14 were published in international journals. When divided into the decade periods, 21 papers (19.4%) between 1990 and 1999 and 26 papers (24.1%) between 2000 and 2009, and 42 papers (48.1%) were published in 2010 and 2018, respectively. When divided into the survey area by a bearing (East sea, South-East sea, South Sea, and West Sea), 65 papers(69.1%) were carried out in the South Sea. Followed by 13 papers (13.8%) in the East Sea and 8 papers (8.5%) in the West Sea and the South-East Sea, respectively. The surveyed area was dominant in Gwangyang Bay (18 papers; 24.3%), followed by Gadeok-do (8 papers; 10.8%). Of the diversity indices, 14 indices were used to calculate the ecological status. The dominant indices were IRI(Index of relative importance), Pi(Prey-specific abundance) followed by E (electivity index) and Bi (dietary breadth index). A total of 11 statistical methods were used to find feeding strategy. The correlation analysis and Bray-Curtis similarity matrix were applied most frequently. In terms of ecological research topics, the papers focused on seasonal or spatial differences before 2000. Since 2000, specifically, the international journal papers have been published based on competition, sympatric, niche overlap and other ecological topics.

Effect of Cardanol Content on the Antibacterial Films Derived from Alginate-PVA Blended Matrix (알지네이트-폴리비닐알콜 블랜드 항균 필름 제조를 위한 카다놀 함량의 영향)

  • Ahn, Hee Ju;Kang, Kyung Soo;Song, Yun Ha;Lee, Da Hae;Kim, Mun Ho;Lee, Jae Kyoung;Woo, Hee Chul
    • Clean Technology
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    • v.28 no.1
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    • pp.24-31
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    • 2022
  • Petroleum-based plastics are used for various purposes and pose a significant threat to the earth's environment and ecosystem. Many efforts have been taken globally in different areas to find alternatives. As part of these efforts, this study manufactured alginate-based polyvinyl alcohol (PVA) blended films by casting from an aqueous solution prepared by mixing 10 wt% petroleum-based PVA with biodegradable, marine biomass-derived alginate. Glutaraldehyde was used as a cross-linking agent, and cardanol, an alkyl phenol-based bio-oil extracted from cashew nut shell, was added in the range of 0.1 to 2.0 wt% to grant antibacterial activity to the films. FTIR and TGA were performed to characterize the manufactured blended films, and the tensile strength, degree of swelling, and antibacterial activity were measured. Results obtained from the FTIR, TGA, and tensile strength test showed that alginate, the main component, was well distributed in the PVA by forming a matrix phase. The brittleness of alginate, a known weakness as a single component, and the low thermal durability of PVA were improved by cross-linking and hydrogen bonding of the functional groups between alginate and PVA. Addition of cardanol to the alginate-based PVA blend significantly improved the antibacterial activity against S. aureus and E. coli. The antibacterial performance was excellent with a death rate of 98% or higher for S. aureus and about 70% for E. coli at a contact time of 60 minutes. The optimal antibacterial activity of the alginate-PVA blended films was found with a cardanol content range between 0.1 to 0.5 wt%. These results show that cardanol-containing alginate-PVA blended films are suitable for use as various antibacterial materials, including as food packaging.

Effects of Kyejiinsam-tang in MIA-Induced Osteoarthritis Rats (계지인삼탕(桂枝人蔘湯)이 MIA로 유도된 골관절염 유발 Rat에 미치는 영향)

  • An, Soon-Sun;Heo, Dong-Seok
    • The Journal of Korean Medicine
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    • v.34 no.3
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    • pp.69-85
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    • 2013
  • Objectives: This study investigated the anti-osteoarthritic effects of Kyejiinsam-tang (hereinafter referred to KIT) on the monosodium iodoacetate (MIA)-induced osteoarthritis rats. Methods: Anti-oxidative effects of KIT were measured by scavenging activities of DPPH, reactive oxygen species (ROS) and nitric oxide (NO). Scavenging activities of anti-oxidation in lipopolysaccharide (LPS)-treated RAW 264.7 cells were also measured for inhibitory effects against the production of inflammatory mediators (tumor necrosis factor-${\alpha}$, interleukin-$1{\beta}$, interleukin-6). Osteoarthritis was induced in rats by injecting MIA in the knee joint. Rats were divided into a total of 4 groups (n=6). The normal group were not treated at all without inducing osteoarthritis whereas the control group were induced for osteoarthritis by MIA and oral medicated physiological saline per day. The positive comparison group was injected with MIA and after 7 days, 2 mg/kg of Indomethacin. The experimental group was injected with MIA and after 7 days was medicated with 34 mg/kg of KIT. Indomethacin and KIT were orally-medicated for each substance a total of 4 weeks, once per day. Weight-bearing on hind legs was measured every week after MIA injection. At the end of the experiment (5 weeks after MIA injection), micro CT (computed tomography)-arthrography and histopathological examinations on the articular structures of knee joint were performed. The effect on inflammatory cytokines and immunological cells in synovial fluid was measured. Volume of cartilage was measured by micro CT-arthrography. Injury to synovial tissue was measured by H & E (hematoxylin and eosin), Safranin-O immunofluorescence. Results: 1. Cytotoxicity against hFCs was insignificant. 2. KIT showed the potent full term for DPPH. 1. NO was significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and ROS was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 2. IL-6 and IL-$1{\beta}$ were significantly reduced by KIT (at 100, $200{\mu}g/m{\ell}$) and TNF-${\alpha}$ was also reduced, but not significantly, by KIT (at $200{\mu}g/m{\ell}$). 1. In hind legs weight-bearing measurement, level of weight increased. 2. Functions of liver and kidney were not affected. 3. IL-$1{\beta}$ was significantly reduced and TNF-${\alpha}$, IL-6 were also reduced but not significantly. 4. PGE2 (prostaglandin E2), LTB4 (leukotriene B4) were significantly reduced in the KIT group. 5. MMP-9 (matrix metalloproteinase-9), TIMP-1 (tissue inhibitor of metalloproteinases-1) and Osteocalcin were significantly reduced in the KIT group. 6. Destruction of cartilage on micro CT arthrography was reduced but had no significant differences. 7. Histopathologically, injury to synovial membrane of the KIT group was decreased and proteoglycan content of KIT group was increased. Conclusions: According to this study, Kyejiinsam-tang has inhibiting effect on the progression of arthritis in MIA-induced osteoarthritis rat. Kyejiinsam-tang has anti-oxidants and anti-inflammation effects, and is related to inhibiting the activity of inflammatory cytokine and injury of volume in cartilage.