• BMB Reports
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• v.38 no.4
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• pp.414-419
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• 2005

The production of recombinant antibodies has been generally recognized as time-consuming and labor-intensive. The aim of our study is to construct mammalian expression vectors containing the cDNA encoding the human constant regions and murine variable regions to massively and cost-effectively produce full-length chimeric antibodies. Unique restriction sites flanking the Ig variable region were designed to allow for the replacement of variable regions generated by PCR. Western blot analysis of the chimeric antibodies revealed that the expressed products were of the predicted size, structure and specificity. The usefulness of the vectors was confirmed by construction of human-mouse chimeric antibody-HCAb which secretes murine antibody against the human colorectal cancer. Selected in medium containing gradually increasing methotrexate (MTX), clones with increased expression of the product gene can be efficiently generated. The secretion of recombinant chimeric antibody-HCAb yielded $30\;pg\;cell^{-1}\;day^{-1}$ at $10^{-6}\;M$ MTX. With this high-level expression from pools, the convenient and rapid production of over 100 milligram amounts per liter of recombinant antibodies may be achieved, which indicates the significant roles of pYR-GCEVH and pYR-GCEVL in the production of chimeric antibodies.

• The Journal of Korean Medicine
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• v.31 no.2
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• pp.19-35
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• 2010

Objectives: This study was carried out to find the effects of Changchuldoin-tanggamibang (hereinafter referred to CDIT) on the inhibition of arthritis induced by collagen on DBA/1J mouse. Methods: The experimental mice were divided into four groups: normal group (Nr), control group (CIA-CT), methotrexate group (CIA-MTX), and Changchuldoin-tanggamibang group (CIA-CDIT). Cytotoxicity, hepatotoxicity, arthritis index, value of immunocytes in draining lymph node and paw joint, and rheumatoid factor (IgG, IgM) in serum were measured in vivo. Results: 1. Cytotoxicity against hFCs was not shown in any concentration. 2. Hepatotoxicity was low in the CDIT-treated group compared with the MTX group. 3. The arthritis index decreased significantly. 4. In total cell counts of DLN and paw joint, the cells in DLN increased significantly while there was a significant decrease in paw joint. 5. In lymph nodes, CD19+, CD3+, CD4+, CD8+, CD3+/CD8+, CD3+/CD69+, CD4+/CD25+, CD3+/CD49b+, and CD4+/CD44+ cells increased significantly, while B220+/CD23+, and CD11c+/MHCII+ cells decreased significantly. 6. In joints, CD3+, CD4+, CD4+/CD25+, and CD11b+/Gr-1+ cells decreased significantly. 7. The level of IgG decreased and the level of IgM significantly decreased compared with the control. 8. Anti-collagen II in serum decreased compared with the control. 9. Around the joint of the CDIT group, infiltration of inflammation, synovial hyperplasia, invasion of cytokine, of cartilage, deposition of collagen and synovial injury decreased compared with the control in histopathologic observation (HE, MT staining). Conclusions: Comparison of the results for this study showed that CDIT had immunomodulatory effects. We expect that CDIT could be used as a effective drug for not only rheumatoid arthritis but also another auto-immune diseases. Therefore, we have to survey continuously, looking for effective substances and mechanisms in the future.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.4
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• pp.1140-1146
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• 2004

This study was carried out to investigate the comparison of immunomodualtory effects of water-extracted Ginseng Radix(GR), Pilose Asia-bell(PA), Astragali Radix(AR), Astractylodes Rhizoma alba(AA) and Dioscoreae Rhizoma (DR). The parameter examined to assess apparent immunomodulatory effect of the water-extracted GR, PA, AR, AA and DR included the regulation of Nitric oxide (NO), the expression of Th1/Th2 type cytokine, the change of B cell phenotype. The water-extracted GR, PA, AR, AA and DR inhibited NO production and iNOS protein expression in LPS stimulated RAW 264.7 macrophage cells. In the Th1 and Th2 cytokine expression, the water-extracted GR, PA, AR, AA and DR induced IL-2 and IFNr mRNA gene expression. Therefore, it seems that the water-extracted GR, PA, AR, AA and DR have a inducing effect of Th1 type cytokines. In the Flow cytometry analysis, the water-extracted GR, PA, AR, AA and DR did not change B cell phenotype (CD45R/B220). The water-extracted GR, PA, AR, AA and DR have a reducing effect of immune suppression cause by Methotrexate (MTX), an agent of immune suppression. These results suggest that the immunomodulatory effects of the water-extracted GR, PA, AR, AA and DR may be, in part, associated with the inducing IL-2 and IFNr mRNA gene expression in and regulation of NO production in macrophage cells.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.4
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• pp.1000-1010
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• 2005

This study was carried out to investigate the comparison of immunomodualtory effects of water-extracted Aconiti lateralis Preparata Radix(PR), Zingiberis Rhizoma(ZR), Cinnamomi Cortex(CC) and Evodiae Fructus(EF). The parameter examined to assess apparent immunomodulatory effect of the water-extracted PR, ZR, CC and EF included the regulation of Nitric oxide (NO). Also, ZR and EF represent the expression of Th1/Th2 type cytokine, the change of B cell phenotype. The water-extracted PR, ZR, CC and EF inhibited NO production and iNOS protein expression in LPS stimulated RAW 264.7 macrophage cells. In the Th1 and Th2 cytokine expression, the water-extracted ZR and EF induced IL-2, IFNr and IL-10 mRNA gene expression. Therefore, it seems that the water-extracted ZR and EF have a inducing effect of Th1 and Th2 type cytokines. In the Flow cytometry analysis, the water-extracted ZR and EF changed B cell phenotype (CD45R/B220), did NOT in PR and CC. The water-extracted PR, ZR, CC and EF have a reducing effect of immune suppression cause by Methotrexate (MTX), an agent of immune suppression. These results suggest that the immunomodulatory effects of the water-extracted ZR and EF may be, in part, associated with the inducing IL-2 and IFNr mRNA gene expression In and regulation of NO production in macrophage cells.

• The Journal of Pediatrics of Korean Medicine
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• v.23 no.1
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• pp.73-83
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• 2009

Objectives This study is to report a case that has an important meaning as a result of treating Juvenile Rheumatoid Arthritis. We investigated a patient who had to maintain his life with western medicines such as DMARDs, NSAIDs and steroids for a long time. The patient has recovered from all symptoms and his ESR, CRP has been back to normal range with oriental medicine treatment. Methods The patient had fever, especially repeated fever during the afternoon, pain and swelling of joints, generalized skin eruption, anorexia, delayed growth, weight loss, fatigue. So we treated him with herbal medicine and reduced his western medicine. The aim of treatment was recovery from Juvenile Rheumatoid Arthritis after discontinuance of all western medicine. Results The symptoms of systemic type Juvenile Rheumatoid Arthritis was vanished and the patient maintains his condition with oriental medicine treatment after stopped all DMARDs such as MTX(methotrexate) and NSAIDs. His ESR and CRP levels were back to the normal range. After this treatment the patient's height and weight has been increased which showed a significant meaning in growth to the child. Conclusions This study showed that oriental medicine can elevate the Juvenile Rheumatoid Arthritis patient's quality of life with continuous health care and treatment for major problem. For more accurate studies, further studies would be needed with more cases.

• Proceedings of the Materials Research Society of Korea Conference
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• 2011.05a
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• pp.52.2-52.2
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• 2011

Flat form technology for constructing anticancer loaded multi-walled carbon nanotubes (mwCNTs) was introduced in this study. Conventional anticancer drugs, such as MTX (Methotrexate), cisplatin, DOX (Doxorubicin hydrochloride), DAU (Daunorubicin) and EPI (epirubicin) were bio-conjugated with folic acid (FA) for selective targeting tumor cells. Loading efficiencies of the used anticancer drugs on mwCNTs have shown different order of bindings depending on the molecular bind affinity of NH (amine) formation on mwCNTs. MTT assays have shown increased selective target efficiency of FA conjugated mwCNTs on breast cancer cell growth inhibition. All results collectively indicated promising application of mwCNTs as a smart inorganic nanomaterial for selective targeting drug delivery vehicle at tumor tissues.

• Journal of Korean Medicine Rehabilitation
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• v.19 no.3
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• pp.135-146
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• 2009

Objectives : The purpose of this study is to report the effect of live leech(hirudo medicinalis) therapy with oriental medication on the patient with regional rheumatoid arthritis. The patient had resistant to DMARDs so she was in the debilitating condition and suffered from edema, pain, stiffness of joint and purpura before live leech(hirudo medicinalis) therapy. Methods : The regional rheumatoid arthritis patient had edema, pain, stiffness of joint and purpura on right foot. We treated the patient with live leech(hirudo medicinalis) therapy and oriental medicine. Results : The symptoms of regional rheumatoid arthritis was improved through live leech(hirudo medicinalis) therapy and oriental medicine. The patient maintains her condition with oriental medicine treatment after stopped all DMARDs such as MTX(methotrexate) and NSAIDs. After this treatment the patient's walking difficulty has been improved. Conclusions : This study shows that live leech(hirudo medicinalis) therapy and oriental medicine can elevate the regional rheumatoid arthritis patient's quality of life with continuous health care and treatment for major problem. For more accurate studies, further studies would be needed with more cases.

• The Korean Journal of Zoology
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• v.20 no.4
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• pp.159-168
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• 1977

In order to evaluate the mutagenic potentential in mammalian system for those pesticides which were proved to be mutagenic in Salmonella microsome assay system, we have studied drug-resistant mutagenesis in cultured L5178Y cells and unscheduled DNA synthesis in human lymphocytes in vitro. We have tested five pesticides: insecticides DDVP and trichlorfon, fungicide TMTD and herbicides MO and NIP. Of these pesticides, TMTD induced weak mutation to MTX-resistance in L5178Y cells in vitro and gave positive responses in DNA repair assay system. Therefore, its potential genetic risks in human beings should be re-evaluated. DDVP and trichlorfon gave negative response in L5178Y mutagenesis test system but stimulated incorporation of $^{3}H$-TdR in DNA repar assay. MO and NIP gave also negative responses both in L5178Y mutagenesis test systemand in DNA repair assay system.

• Journal of the Korean Chemical Society
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• v.37 no.1
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• pp.131-135
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• 1993

A new series of 2-desamino, 2-desamino-2-methyl, and 2-amino analogues of aminopterin intermediates (7a), (7b), and (7c) were synthesized from 2-amino-3-cyano-6-chloromethylpyrazine. These derivatives were synthesized via two step sequence. Thus, cyclization of 2-amino-3-cyano-6-[(S-p-carbenzyloxyphenyl)thiomethyl]-pyrazine(14) with formamidine HCl, acetamidine HCl or guanidine HCl provided condensed compounds (15a), (15b) which upon base hydrolysis yielded the desired products (7a), (7b) and (7c).

• Journal of Sasang Constitutional Medicine
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• v.9 no.1
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• pp.315-337
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• 1997

In order to compare the dffects of $Galg{\breve{u}}nhaegit'ang$(葛根解肌湯) of "Dongeuisusebowon(東醫壽世保元)and Won's(元)-$Galg{\breve{u}}nhaegit'ang$(葛根解肌湯) of "Dongeuisasansinpyun(東醫四象新編)" on the immune respone, Sprague-Dawley male rats were used and randomly divided into four groups. Normal group was under normal condition, Control group was injected i.v. with 2mg/kg Methotrexate(MTX) on the 9th day and 11th day after sensitization with SRBC on the 5th day, $Galg{\breve{u}}nhaegit'ang$ group was fed with 1ml of $Galg{\breve{u}}nhaegit'ang$ and Won's-$Galg{\breve{u}}nhaegit'ang$ group was fed with 1ml of Won's-$Galg{\breve{u}}nhaegit'ang$ by oral during eighteen days. In the 9th day and the 11th day after oral feeding with medication, MTX was injected in tail of rats in order to reduce immune function. Leukocyte count, lymphocyte ratio, lymphocyte count of spleen, lymphocyte count of bone marrow, contact hypersensitivity to DNFB, morphologic change of thymus cell, and electropherogram of serum protein were estimated and compared according to the group. The results are as follows : 1. Before and after MTX injection, leukocyte(WBC) count was increased signigicantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 2. Before and after MTX injection, lymphocyte ratio was not significantly different in Won's-$Galg{\breve{u}}nhaegit'ang$ group and in $Galg{\breve{u}}nhaegit'ang$ group compared to control group. 3. The lymphocyte count of spleen was increased significantly in $Galg{\breve{u}}nhaegit'ang$ group compared to control group and Won's-$Galg{\breve{u}}nhaegit'ang$ group. Won's-$Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 4. The lymphocyte count of bone marrow was increased significantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group and $Galg{\breve{u}}nhaegit'ang$ group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control group. 5. Contact hypersensitivity was increased significantly in Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to other group. $Galg{\breve{u}}nhaegit'ang$ group had no significant difference compared to control groups. 6. In the morphologic change of thymus cell, control group compared to normal group had a indistinct boundary between cortex and medulla and lymphocyte cell density of thymus was low. $Galg{\breve{u}}nhaegit'ang$ group and Won's-$Galg{\breve{u}}nhaegit'ang$ group compared to control group had a definite boundary between cortex and medulla and lymphocyte cell density of thymus was high. 7. In the SDS-PAGE electropherogram of serum protein, Won's-$Galg{\breve{u}}nhaegit'ang$ group had a wide band of nearby 25,000 Dalton, and which meant IgG generated more actively. Considering this results, $Galg{\breve{u}}nhaegit'ang$ group and Won's-$Galg{\breve{u}}nhaegit'ang$ group have an effect on the depression of immune function induced by MTX, and especially Won's-$Galg{\breve{u}}nhaegit'ang$ group has an significant effect than $Galg{\breve{u}}nhaegit'ang$ group.