• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.121-129
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• 2002

Using recombinant Chinese hamster ovary (CHO) cells, strategies for developing high producers for the recombinant human Transforming Growth $Factor-{\beta}1$ ($TGF-{\beta}1$) protein are proposed and their physiological characteristics in cell cultures were investigated. $TGF-{\beta}1$ is a pleiotrophic polypeptide involved in various biological activities, including cell growth, differentiation, and deposition of extracellular matrix proteins. The CHO cells included human $TGF-{\beta}1$ cDNA in conjunction with a dihydrofolate reductase (DHFR) gene, which was cotransfected into the cells to amplify the transfected $TGF-{\beta}1$ cDNA. As a first-round screening of the transfected cells, a relatively high $TGF-{\beta}1$-producing cell line was selected, and then, it acquired a resistance to increasing concentrations of methotrexate (MTX) up to $60{\mu}M$,resulting in a significant improvement in its $TGF-{\beta}1$ biosynthetic ability. After applying a monoclonal selection strategy to the MTX-resistant cells, more productive cells were screened, including the APP-3, App-5, and App-8 cell lines. These high producers were compared with two other cell lines (AP-l cell line without amplification of transfected $TGF-{\beta}1$ cDNA and nontransfectant of $TGF-{\beta}1$ cDNA) in terms of cell growth, $TGF-{\beta}1$ productivity, sugar uptake, and byproduct formation, in the presence or absence of MTX in the culture medium. Consequently, both monoclonal selection as well as an investigation of the physiological characteristics were found to be needed for the efficient screening of higher $TGF-{\beta}1$ producers, even after the transfection and amplification of the transfected gene.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.759-766
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• 2003

Recombinant Chinese hamster ovary (rCHO) cell lines expressing a high level of human thrombopoietin (hTPO) or its analog, TPO33r, were obtained by transfecting expression vectors into dihydrofolate reductase-deficient (dhfr) CHO cells and subsequent gene amplification in media containing stepwise increments in methotrexate (MTX) level such as 20, 80, and 320 nM. The parental clones with a hTPO expression level $>0.40\;{\mu}g/ml$ (27 out of 1,200 clones) and the parental clones with a TPO33r expression level $>0.20\;{\mu}g/ml$ (36 out of 400 clones) were subjected to 20 nM MTX. The clones that displayed an increased expression level at 20 nM MTX were subjected to stepwise increasing levels of MTX such as 80 and 320 nM. When subjected to 320 nM MTX, most clones did not display an increased expression level, since the detrimental effect of gene amplification on growth reduction outweighed its beneficial effect of specific TPO productivity ($q_{TPO}$) enhancement at 320 nM MTX. Accordingly, the highest producer subclones ($1-434-80^{*}$ for hTPO and $2-3-80^{*}$ for TPO33r), whose $q_{TPO}$ was 2- to 3-fold higher than that of their parental clones selected at 80 nM MTX, were isolated by limiting dilution method and were established as rCHO cel1 lines. The $q_{TPO}$ of $1-434-80^{*}\;and\;2-3-80^{*}\;was\;5.89{\pm}074\;and\;1.02{\pm}0.23\;{\mu}g/10^6$ cells/day, respectively. Southern and Northern blot analyses showed that the enhanced $q_{TPO}$ of established rCHO cell lines resulted mainly from the increased TPO gene copy number and subsequent increased TPO mRNA level. The hTPO and TPO33r produced from the established rCHO cell lines were biologically active in vivo, as demonstrated by their ability to elevate platelet counts in treated mice.

• Journal of Preventive Medicine and Public Health
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• 제20권1호
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• pp.1-9
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• 1987

항암제와 세포독성과의 관계를 알아보고자 인혈배양 임파구에서 SCE빈도, 세포분열지수 및 세포분열주기변동을 조사한 결과는 다음과 같다. 1) 항암제 농도증가에 따라 SCE빈도는 대조군에 비해 유의하게 증가되었으며 (P<0.01), methotrexate에서는 유의성이 인정되지 않았다(P>0.05). 2) 세포분열지수는 cyclophosphamide를 제외한 공시 항암제에 대하여 현저하게 감소되었다. 3) 항암제의 농도증가에 따라 세포분열주기는 현저하게 지연되었으며 고농도에서는 현저한 세포분열 억제로 정상적인 세포분열이라 할 수 없는 강한 세포독성효과를 보였다. 이 결과는 알킬계 약제가 다른 항암제에 비해 강한 SCE 유발제이며 SCE유발과 세포분열주기 지연과는 깊은 상관성이 인정되지만 methotrexote경우에서는 그 상관성을 인정할 수 없었다. 이는 SCE유발과 세포분열주기지연이 서로 다른 기전에 의해서도 나타남을 제시해주고 있다.

• 동의생리병리학회지
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• 제16권3호
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• pp.483-489
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• 2002

APA-01, which is an aqueous extract of five Chinese herbs, is a modified formula of Huoxiang-Zhengqi-San. The effect of new herb extract on immune response was investigated. The parameter examined to assess apparent immune response of APA-01 in mice included changes of body weight, relative weight of immune organs, cell proliferation and cytokine gene expression. The body weight and relative weight of immune organs were not significantly changed among the tested groups. In the spleen cell prolijeration assay, APA-01 increased the cell proliferation in a dose-dependent manner. Methotrexate (MTX), an agent of immune suppression, inhibited the spleen cell proliferation (IC/sub 50/: 800㎍/㎖). However, APA-01 significantly inhibited the suppression of mouse spleen cell proliferation. Therefore, it seems that APA-01 has a reducing effect of immune suppression. Immunomodulatory effect of APA-01 was further investigated using reverse transcription polymerase chain reaction (RT-PCR) in mouse spleen cells. In RT-PCR test, APA-01 enhanced the expression of cyclooxygenase-2 (COX-2) mRNA in a dose-dependent manner. In spite of immune suppression by MTX, COX-2 mRNA was induced by co-treatment with APA-01. These results suggest that APA-01 stimulates the proliferation of spleen cells, regulates the expression of COX-2 mRNA, and accelerates the recovery of inhibition of spleen cell proliferation induced by MTX, thus providing the immunological basis for clinical benefit of APA-01.

• Tuberculosis and Respiratory Diseases
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• 제58권1호
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• pp.83-88
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• 2005

Leflunomide는 최근에 개발된 항류마티스 약물로서 기존의 약물과 작용기전이 다르면서 효과가 우수하여 활동성 류마티스관절염에 널리 사용되고 있다. 알려진 부작용에는 간독성과 피부독성, 조혈기능부전, 위장관계 증상등이 있는데, 시판후 조사에서는 호흡기계 부작용도 드물지만 경한 정도에서 치명적인 경우까지 모두 보고 되었다. 세계 각 처에서의 시판후 조사에 의하면 호흡기계 부작용으로 사망한 경우가 있어서 심각한 폐독성의 가능성을 시사한다. 우리나라에서는 leflunomide로 인한 호흡기계 부작용에 대한 보고가 없었다. 저자들은 leflunomide에 의한 간질성 폐렴 1예를 경험하였기에 문헌고찰과 함께 보고하는 바이다.

• Radiation Oncology Journal
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• 제38권1호
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• pp.35-43
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• 2020

Purpose: This retrospective study compares higher-dose whole-brain radiotherapy (hdWBRT) with reduced-dose WBRT (rdWBRT) in terms of clinical efficacy and toxicity profile in patients treated for primary central nervous system lymphoma (PCNSL). Materials and Methods: Radiotherapy followed by high-dose methotrexate (HD-MTX)-based chemotherapy was administered to immunocompetent patients with histologically confirmed PCNSL between 2000 and 2016. Response to chemotherapy was taken into account when prescribing the radiation dose to the whole brain and primary tumor bed. The whole brain dose was ≤23.4 Gy for rdWBRT (n = 20) and >23.4 Gy for hdWBRT (n = 68). Patients manifesting cognitive disturbance, memory impairment and dysarthria were considered to have neurotoxicity. A median follow-up was 3.62 years. Results: The 3-year overall survival (OS) and progression-free survival (PFS) were 70.0% and 48.9% with rdWBRT, and 63.2% and 43.2% with hdWBRT. The 3-year OS and PFS among patients with partial response (n = 45) after chemotherapy were 77.8% and 53.3% with rdWBRT, and 58.3% and 45.8% with hdWBRT (p > 0.05). Among patients with complete response achieved during follow-up, the 3-year freedom from neurotoxicity (FFNT) rate was 94.1% with rdWBRT and 62.4% with hdWBRT. Among patients aged ≥60 years, the 3-year FFNT rate was 87.5% with rdWBRT and 39.1% with hdWBRT (p = 0.49). Neurotoxicity was not observed after rdWBRT in patients aged below 60 years. Conclusion: rdWBRT with tumor bed boost combined with upfront HD-MTX is less neurotoxic and results in effective survival as higher-dose radiotherapy even in partial response after chemotherapy.

• 한국보건간호학회지
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• 제16권1호
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• pp.176-189
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• 2002

The purpose of this study were to illustrate the various medication error types and causes and identified to related drugs to provide basic data for preventing nurses' medication error by analysing 73 cases of AJN 'medication Error' column(1993, Oct -2000, Nov). Nurses' types of medication error were classified into 7 types. The most frequent error types are wrong medication$(21.9\%)$ and the wrong dose$(21.9\%)$ together. The others are wrong $time(4.1\%)$, $omission(2.7\%)$, mechanical $error(2.7\%)$, incorrect IV $rate(1.4\%)$. wrong route $administration(1.4\%)$ in order. Nurses' causes of medication error were 9 kinds. The most frequent type is confusing between similar drug shape, color, size, name, injection devices and patient's $name(43.9\%)$ and the others are lack of knowledge about $drugs(26.8\%),\; slips(7.3\%),\; miscalculating\;dose(4.9\%)$, incorrect adjusts $devices(4.9\%)$, difficulty to read or illegible decimal $point(4.9\%),$ $abbreviation(2.4\%)$, fatigue with $overwork(2.4\%)$ and no communication with $patient(2.4\%)$ in order. Related drugs with medication error are as follows. - dose unit(IU. minims. mcg/min. mEq) : Heparin. insulin. synthetic calcitonin, some enzymes and hormones, vitamins, some antibiotics, tuberculin injection. MgSO4 injection. nitroglycerin - similar size, color and shape drug : $0.9\%$ N/S and acetic acid $0.25\%$ for irrigation. premixed 2mg lidocaine sol. and $0.9\%$ N/S, gentamycin 20mg/2mL for children and 80mg/2mL for adult, dextroamphetamine 5mg and 10mg capsule. sedatives chloral hydrate 250mg/5mL and 500mg/5mL - similar name :Aredia(pamidronate disodium) and Adriamycin(doxorubicin), Lamictal (lamotrigine) and Lamisil 250mg. Elderpryl and enalapril, cefotaxime and cefoxitin, carboplatin and cisplatin, sumatriptan and zolmitriptan, Celebrex and Celexa, Humulin and Humalog, Percodan and Percocet, Diabeta and Diabinese, Epivir and Retrovir, Xanax(alprazolam) and Zantac(ranitidine) - decimal point : low molecular weight warfarin, methotrexate - unfamiliar drug uses of familiar drug ; methotrexate. droperidol, imipramine, propranolol - number of drug name(misleading chemical name) : 6-thioguanine, 6-mercaptopurine, 5-fluorouracil - type of administration route : Oxycodone(OxyContin). - administration time : acarbose(Precose). - injection way (Z-track method): hydroxyzine - epidural cathether : LMWHs(enoxaparin, dalteparin), - ADD Vantage self contained delivery system : ceftriaxone(Rocephin)

• 한국동물학회지
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• 제38권4호
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• pp.565-569
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• 1995

분열형 호모인 Schizosaccbarornyces pombe에서 RecA 유사 단백질은 hydroxyurea (HU)나 methotrexate (MTX)와 같은 nucleotide pool 형성에 관여하는 효소의 저해제에 의해 유도발현된다. 본 연구에서는 대장균의 RecA 단백질과 S. pombe의 ribonucleotide reductase가 구조적으로 유사한지를 RecA 항체를 이용한 면역침전법과 [5-$^3$H]CDP를 기질로하는 thin layerchromatography방법을 통하여 알아보고자 하였다. S. pombe의 rinonucleotide reductase활성은 100mM HU에 의해 대조군에 비해 26.3% 저해되었으며, RecA 항체를 이용한 면역침전에 의해서도 43.3% 저해되었다. 이와 같은 결과는 S. pombe의 rinonucleotide reductase가 대장균의 RecA 단백질과 구조적으로 유사함을 시사하는 것이다.

• Radiation Oncology Journal
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• 제7권2호
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• pp.269-277
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• 1989

1979년 8월부터 1986년 12월까지 관해유도 치료 후 골수 관해(M1)가 온 134예의 소아 급성 임파구성 백혈병 환아가 18Gy또는 24Gy의 전뇌 방사선 조사와 척수강내 methotrexate주입의 예방적 중추신경계 치료를 받았다. 중추신경계에서 처음 재발한 환자는 14.2%(19/134)였고 이중 증추신경계에서만 재발한 예는 5.2%(7/134), 골수 또는 고환에서의 재발과 동시에 재발한 예는 9%(12/134)였다. 남아와 연령이 많을수록 중추신경계 재발이 높았으며 진단당시 말초혈액내의 백혈구수가 50,000/ul이상인 예의 재발율이 높았다. 방사선 조사량 24Gy에서의 중추 신경계 재발율이 18Gy인 경우보다 높았으나 유의하지는 않았다. 전체 중추신경계 재발예 중 89%가 예방적 중추신경계 치료개시 후 4년 이내에 발생하였으며, 중추신경계에서만 재발예는 100%가, 중추신경계와 골수 또는 고환의 재발과 동시에 재발한 예의 83%가 각각 관찰되었다. 재발의 위험이 있는 환자를 감안한전체 중추신경계 재발율은 유지 화학요법 기간 중에는 11.9%였고 유지 화학 요법 종료 이후의 기간에서는 4.9%였다.

• 대한본초학회지
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• 제34권2호
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• pp.41-47
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• 2019

Objectives : A traditional herbal prescription, Kyung-Ok-Ko (KOK), has long been used in oriental medicine as an invigorant for age-related diseases, such as amnesia and stroke. However, the beneficial value of KOK for immune responses is largely unknown. Based on the above mentioned effects of KOK, other previous reports, and its use in traditional medicine, we hypothesized that KOK displays beneficial effects against methotrexate (MTX)-induced immune suppression. Methods : We investigated the effects of KOK (0.6 g/kg/day, oral (p.o.)) on deteriorated immunity caused by MTX (2 mg/kg/day, p.o.) in an immune suppression mouse model. MTX was fed to mice once a day for 7 days. After the immune responses of the mice deteriorated by MTX treatment, KOK in water was fed to the mice once a day for 14 days. We then measured the expression levels of various cytokines, such as T helper cell (Th1, Th2) cytokines, and the number of immune cells, such as spleen T cells, B cells, and macrophages. Results : The data showed that MTX decreased Th1 profiles (interferon $(IFN)-{\gamma}$, interleukin (IL)-2, IL-12) and the number of immune cells, and increased Th2 profiles (IL-4, IL-5, IL-13), which were normalized significantly by post-administration of KOK. However, there was no significant difference in body-weight gain between MTX- and KOK-treated mice. Conclusion : These results indicate that KOK has immune-enhancing functions and reduces immunotoxicity of MTX, suggesting that supplementation with KOK will improve immune responses clinically and be useful for the prevention of immune-related diseases.