• Archives of Pharmacal Research
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• 제21권1호
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• pp.46-53
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• 1998

When NIH3T3 cells were exposed to mild heat and recovered at $37^{\circ}C$ for various time intervals, they were thermotolerant and resistant to subsequent stresses including heat, oxidative stresses, and antitumor drug methotrexate which are apoptotic inducers. The induction kinetics of apoptosis by stresses were determined by DNA fragmentation and protein synthesis using $[35^S]$methionine pulse labeling. We investigated the hypothesis that thermotolerant cells were resistant to apoptotic cell death compared to control cells when both cells were exposed to various stresses inducing apoptosis. The cellular changes in thermotolerant cells were examined to determine which components are involved in this resistance. At first, the degree of resistance correlates with the extent of heat shock protein synthesis which were varied depending on the heating times at $45^{\circ}C$ and recovery times at $37^{\circ}C$after heat shock. Secondly, membrane permeability change was observed in thermotolerant cells. When cells prelabeled with $[^{3}H]$thymidine were exposed to various amounts of heat and recovered at $37^{\circ}C$ for 1/2 to 24 h, the permeability of cytosolic $[^{3}H]$thymidine in thermotolerant cells was 4 fold higher than that in control cells. Thirdly, the protein synthesis rates in thermotolerant and control cells were measured after exposing the cells to the same extent of stress. It turned out that thermotolerant cells were less damaged to same amount of stress than control cells, although the recovery rates are very similar to each other. These results demonstrate that an increase of heat shock proteins and membrane changes in thermotolerant cells may protect the cells from the stresses and increase the resistance to apoptotic cell death, even though the exact mechanism should be further studied.

• 동의생리병리학회지
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• 제19권4호
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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.

• 동의생리병리학회지
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• 제21권2호
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• pp.414-424
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• 2007

Adenophorae Radix (AR), Liriopis Tuber (LT), Dendrobii Monile (DM), Polygonati Officinalis (PO), Polygonati Rhizoma (PR) have been used to treated a variety condition/diseases in traditional oriental medicine. The present study was conducted to investigate the immunmodulatory effects of the water-extracted AR, LT, DH, PO and PR. In spleen cell proliferation assay, DH was significantly enhanced mitogenic activity compared with control group. In RT-PCR, DH ad PO induced IL-2 and IFNr cytokine gene expression in mouse spleen cells. Methotrexate(MTX), immune supression agent, was significantly inhibited mouse spleen cell proliferation(1600 mg/ml). In spite of MTX treatement, DH and PO sustained the spleen cell proliferation, In the flow cytometry analysis, DH stimulated mouse spleen cells showed an increase in B-cell phenotype (CD45R/B220). The water-extracted DH and PO inhibited NO production and iNOS expression in LPS-stimulated RAW 264.7 macrophage cell. DH induced IL-2 and IFNg gene expression in human peripheral mononuclear cells. The GC-MS analysis show that the main component of water-extracted DH was b-Nitroethyl alcohol. The main components of water-extracted PO were Dipirartril-tropico, Methyl sulfoxide and Demsodrox. These data suggest that among these extracts, DH has a protective effcet of immune suppression caused by MTX. DH may be enhance cellular and humoral immune response by the regulation of cytokine gene expression, NO production and B cell production.

• 생명과학회지
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• 제20권5호
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• pp.789-793
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• 2010

복잡한 진핵생물에서의 물리적 지도 작성이나 기능해석에 효모인공염색체(YAC)를 이용하기 위해서는 원하는 target region의 인공염색체화 및 single-copy인 YAC의 복제수를 늘이는 것이 요구된다. 본 연구에서는 YAC manipulation system에 복제수 증폭시스템(copy number amplification system)을 도입한 Simultaneous YAC Manipulation-Amplification (SYMA) system을 구축하였다. 식물염색체를 가진 YAC clone의 splitting과 증폭을 위해 conditional centromere와 thymidine kinase (TK) 유전자를 가진 pBGTK plasmid를 구축하였고, splitting fragment의 PCR을 위한 주형으로 사용하였다. 590 kb의 YAC clone은 splitting과 동시에 copy number amplification element를 가진 100 kb YAC와 490 kb YAC로 분리되었고, 100 kb YAC는 유도기질로 3 mg/ml sulfanilamide와 $50\;{\mu}g/ml$ methotrexate (S3/M50)의 첨가에 의해 14.4배로 그 복제수가 증가하였음을 확인할 수 있었다.

• 대한약침학회지
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• 제3권2호
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• pp.79-97
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• 2000

Objectives : To investigate the effects of herb-acupunctures of Qi tonification herbs or formula on the rat immune depression induced by an anticancer drug, methotrexate (MTX) Methods: Animals were divided into 5 groups; Normal control group was not given any treatment. Immune depression was induced by oral administration of 1mg/kg MTX b.i.d for 4 days in Control, Sample I, II, and III groups. We treated on CV 4 (Guanyuan) with saline, Bojoongiggi-tang (Buzhongyiqi-tang), Ginseng Radix and Astragali Radix herb-acupunctures in Control, Sample I, II, and Ill groups, respectively. In both blood and spleen, the assessment of CD4+ T-cell count, CD8+ T-cell count, CD4/CD8 T-cell ratio was performed. Results: Here we show that 3 herb- acupuncture groups have an influence, to some extent, on CD4+ and CD8+ T-cell counts and CD4/CD8 T-cell ratio in both blood and spleen. Astragali Radix herb-acupuncture, in particular, was found to have significantly increased CD4+ T-cell count in blood and CD4/CD8 T-cell ratio in blood and spleen. Conclusions: The results of this study suggest that herb-acupunctures of Bojoongiggi-tang (Buzhongyiqi-tang), Ginseng Radix and Astragali Radix may have an influence over rat immune depression induced by MTX.

• 동의생리병리학회지
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• 제20권4호
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• pp.896-901
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• 2006

Zingiberis rhizoma(ZB) has been used to treat a various condition and disease in traditional oriental medicine. The present study was conducted to evaluate the immunomodulatory effect of aqueous-extracted ZB(ZBE) on methotrexate (MTX)-induced immune suppression in mouse spleen cells. In spleen cell proliferation assay, ZBE enhanced mitogenic activity in mouse spleen cells. In RT-PCR, ZBE induced IL-2, IFNr and IL-6 cytokine gene expression in mouse spleen cells. In spite of MTX treatment, IL-2, IFNr and IL-6 gene expressions sustained in MTX treated spleen cells. CD45R/B220, pan B marker was slightly increased in ZBE treated mouse spleen cells. IL-6, B cell tropical cytokine, production was induced by ZBE-treated mouse spleen cells and IL-6 production was sustained on MTX-ZBE co-cultured cells. ZBE administration enhanced suNival of S-180 bearing mouse. These data indicate that ZBE has a protective effect of immune suppression caused by MTX, and ZBE may be enhance cellular and humoral function by regulate cytokine gene expression as well as the mitogenic effect on spleen cells.

• Journal of Gynecologic Oncology
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• 제29권6호
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• pp.89.1-89.8
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• 2018

Objective: Highly effective chemotherapy for patients with low-risk gestational trophoblastic neoplasia (GTN) is associated with almost a 100% cure rate. However, 20%-30% of patients treated with chemotherapy need to change their regimens due to severe adverse events (SAEs) or drug resistance. We examined the treatment outcomes of second-line chemotherapy for patients with low-risk GTN. Methods: Between 1980 and 2015, 281 patients with low-risk GTN were treated. Of these 281 patients, 178 patients were primarily treated with 5-day intramuscular methotrexate (MTX; n=114) or 5-day drip infusion etoposide (ETP; n=64). We examined the remission rates, the drug change rates, and the outcomes of second-line chemotherapy. Results: The primary remission rates and drug resistant rates of 5-day ETP were significantly higher (p<0.001) and significantly lower (p=0.002) than those of 5-day MTX, respectively. Forty-seven patients (26.4%) required a change in their chemotherapy regimen due to the SAEs (n=16) and drug resistance (n=31), respectively. Of these 47 patients failed the first-line regimen, 39 patients (39/47, 82.9%) were re-treated with single-agent chemotherapy, and 35 patients (35/39, 89.7%) achieved remission. Four patients failed second-line, single-agent chemotherapy and eight patients (17.0%) who failed first-line regimens were treated with combined or multi-agent chemotherapy and achieved remission. Conclusions: Patients with low-risk GTN were usually treated with single-agent chemotherapy, while 20%-30% patients had to change their chemotherapy regimen due to SAEs or drug resistance. The second-line regimens of single-agent chemotherapy were effective; however, there were several patients who needed multiple agents and combined chemotherapy to achieve remission.

• Radiation Oncology Journal
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• 제19권4호
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• pp.359-368
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• 2001

목적 : 근침윤성 방광암에서 경요도 절제술 후 화학방사선 병용을 통한 방광보존치료를 시행하여 국내에서는 처음으로 그 예비 결과를 발표한 바 있으며, 장기 추적 관찰 결과를 토대로 본 치료법의 효과를 분석하고자 한다. 대상 및 방법 : 1991년부터 1994년까지 병기 T2 에서 T4NxM0의 근침윤성 방광암을 가진 25명의 환자에 방광보존치료법이 시행되었다. 경요도절제술을 통해 가능한 최대한의 방광 종양을 제거한 후 항암제와 방사선요법을 병용하여 시행하였다. 병용치료는 methotrexate, vincristine, adriamycin, cisplatin (M-VAC) 제제로 3회 전보조화학요법을 먼저 시행한 후 cisplatin이 방사선 조사 첫째 주와 넷째 주에 동시에 투여되는 동시화학방사선 요법이 시행되거나(1군), 동일하게 동시화학방사선 요법이 먼저 시행되고 이후 methotrexate, cisplatin, vinblastin (MCV) 제제로 화학요법을 2회 시행하거나(2군), 또는 동시화학방사선 요법만 시행되었다(3군). 각 군의 환자분포는 1군 4명, 2군 14명, 3군 7명이었다. 방사선 치료는 하루에 1회 1.8 혹은 2 Gy를 $40\~45\;Gy$ 정도까지는 진골반에 조사한 후 방광 전체에 추가로 $9\~10\;Gy$를 추가 조사하고, 다시 방광내 종양 부위에 추가 조사하여 총 $60\~65\;Gy$까지 조사하였다. 추적관찰 기간의 중앙값은 70개월 이었다. 결과 : 본 치료와 연관된 독성은 대부분 Grade 2 이하의 경미한 독성이었으며, Grade 3 이상의 독성은 급성합병증으로 혈액학적 독성 1예와 만성합병증으로 방사선 방광염이 2예가 있었다. 전체 대상환자의 5년 생존율은 $67.3\%$였다. 25명 중 20명$(80\%)$에서 화학방사선 병용요법 후 완전 관해를 얻었다. 또한 전체 생존환자 16명 중 10명$(63\%)$에서 방광을 보존하고 있었으며, 생존율에 영향을 미치는 인자에 대한 다변량분석을 시행하였는데, T-병기(p=0.013), 완전관해 유무(p=0.002)가 통계적으로 유의한 인자였다. 결론 : 본 기관에서 시행되어진 근침윤성 방광암에 대한 방광보존치료법은 기존의 근치적 방광절제술에 비하여 대등한 치료성적을 내는 동시에 $63\%$에서 장기보존이 가능하였다. 따라서 본 치료법이 방광암의 치료에 적극적으로 적용되어야 할 것으로 생각하며 향후 여러 기관이 참여하는 활발한 연구를 통해 한국인에게 가장 적절한 치료법을 개발해야 될 것으로 생각한다.

• Clinical and Experimental Pediatrics
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• 제52권5호
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• pp.581-587
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• 2009

목 적 : 골육종의 항암치료 시 MTX의 혈중농도와 예후의 관련 여부에 대해 논란이 있어왔다. 저자들은 MTX의 혈중농도와 골육종의 치료결과간에 상관관계가 있는지 분석하였다. 방 법 : 원자력병원에서 2003년 1월부터 2005년 12월까지 골육종으로 치료를 받은 환자들 중 종양의 원발부위가 사지이고, 폐 전이가 없으며, 수술 전 고용량 MTX ($12g/m^{2}$), cisplatin ($100mg/m^{2}$), doxorubicin ($60mg/m^{2}$)의 병합항암치료를 2회 받은 52명의 환자들을 선택하여 MTX 혈중농도와 항암치료에 대한 종양조직의 괴사율, 생존율의 상관관계를 분석하였다. 결 과 : 52명의 환자들에게 고용량 MTX 항암치료를 총 204회 시행하였다. MTX의 4시간, 24시간, 48시간, 72시간째 혈중농도는 각각 1292.14, 9.29, 1.73, 0.58 µM 이었다. 최고혈중농도(4시간째 혈중농도)는 종양조직의 괴사율과 관계가 없었지만, 24시간째 MTX 혈중농도가 $3.4{\mu}M$ 이상인 환자들이 그렇지 않은 환자들보다 종양조직의 괴사율이 불량하였다 (90%이상의 조직괴사율을 보이는 환자들의 비율, 23.1% 대 50%). MTX 혈중농도는 무사건 생존율과 역 상관관계가 있었다. 최고혈중농도 ${\geq}1,400{\mu}M$ (61.5 % 대 92.1%), 24시간 혈중농도 $\geq$3.4 µM (72.9% 대 96.2%)인 환자들의 무사건 생존율이 낮았다. 결 론 : MTX의 혈중농도가 높은 환자들이 그렇지 않은 환자들보다 치료결과가 불량하였다. 향후 골육종 환자들에서 MTX 약물역동학에 대한 연구를 시행하여 본 연구결과의 기전을 밝혀야 할 것이다.

• Journal of Plant Biotechnology
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• 제2권3호
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• pp.143-149
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• 2000

An Efficient in vitro regeneration system and an optimized Agrobacterium mediated transformation protocol are described, based on the use of young seedling cotyledons of Capsicum annuum L. Optimal regeneration efficiency can be obtained by cultivating cotyledon explants on media containing 4 mg/L benzyladenine and 0.1 mg/L indolacetic acid. The effect of antibiotics used to eliminate Agrobacteria, as well as the toxic level of some generally used selection agents (kanamycin, geneticin, hygromycin, phosphinotricin and methotrexate) in regenerating pepper tissues were determined. To enable the comparison of different selection markers in identical vector background, a set of binary vectors containing the marker genes for NPTII, HPT, DHFR and BAR respectively, as well as the CaMV 35S promoter/enhancer-GUS chimaeric gene was constructed and introduced into four different Agrobacterium host strains.