• Asian Pacific Journal of Cancer Prevention
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• 제14권8호
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• pp.4883-4889
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• 2013

Background: The most common incident cancer and cause of cancer-related deaths in women is breast cancer. The Myc gene is upregulated in many cancer types including breast cancer, and it is considered as a potential anti-cancer drug target. The present study was conducted to evaluate the Myc (gene and protein) expression pattern in an experimental mammary tumour model in rats. Materials and Methods: Thirty six Sprague Dawley rats were divided into: Experimental group (26 animals), which received the chemical carcinogen N-methyl nitrosourea (MNU) and a control group (10 animals), which received vehicle only. c-Myc oncoprotein and its mRNA expression pattern were evaluated using immunohistochemistry (IHC) and semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR), respectively, in normal rat mammary tissue and mammary tumours. The rat glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as internal control for semi-quantitative RT-PCR. Results: Histopathological examination of mammary tissues and tumours from MNU treated animals revealed the presence of premalignant lesions, benign tumours, in situ carcinomas and invasive carcinomas. Immunohistochemical evaluation of tumour tissues showed upregulation and heterogeneous cellular localization of c-Myc oncoprotein. The expression levels of c-Myc oncoprotein were significantly elevated (75-91%) in all the tumours. Semi-quantitative RT-PCR revealed increased expression of c-Myc mRNA in mammary tumours compared to normal mammary tissues. Conclusions: Further large-scale investigation study is needed to adopt this experimental rat mammary tumour model as an in vivo model to study anti-cancer strategies directed against Myc or its downstream partners at the transcriptional or post-transcriptional level.

• 식물병연구
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• 제23권4호
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• pp.379-382
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• 2017

2016년 3월, 모자이크와 황화 병징을 보이는 명월초 잎에서 CMV를 분리하여 CMV-Gyp로 명명하였다. 기주식물의 생물적 반응과 RT-PCR, PCR-RFLP, 이동단백질 및 외피단백질 유전자의 염기서열 분석을 통해 CMV를 동정하였다. 가지과(담배, 고추, 꽈리)와 비름과(흰명아주, 붉은명아주)에서는 전형적인 CMV의 병징이 나타났으나 박과(쥬키니호박, 오이)에서는 병징이 발현되지 않는 특성을 보였다. 3a 및 CP 유전자의 계통학적 분석 결과, CMV-Gyp가 CMV Subgroup II에 속하는 것으로 나타냈다. CMV-Gyp의 이동단백질과 외피단백질유전자의 서열은 아미노산 수준에서 CMV-Hnt와 99.3% 및 100% 일치하였다. 이 논문은 명월초에서 CMV 감염에 대한 첫 보고이다.

• 동아시아식생활학회지
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• 제7권4호
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• pp.484-490
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• 1997

백삼성분이 탐식세포를 통한 면역반응조절에 어떠한 영향을 미치는가를 알아보기 위해 total saponin이나 ginsenoside Rb$_2$성분이 마우스 복강 탐식 세포의 증식과 nitric oxide(NO)분비 및 NOS 유전자 발현에 미치는 영향을 조사해 보았다. Total saponin 또는 ginsenoside Rb$_2$성분을 탐식세포 배양액에 0-256$\mu\textrm{g}$/$m\ell$ 농도로 첨가하여 배양한 후 $^3$H-thymidine incorporation법으로 분석을 실시한 결과 total saponin은 64$\mu\textrm{g}$/$m\ell$ 농도에서 세포내 DNA 합성을 증가시켰으며, ginsenoside Rb$_2$ 성분의 경우 16$\mu\textrm{g}$/$m\ell$ 또는 64$\mu\textrm{g}$/$m\ell$ 농도에서 DNA 합성이 증가되었다. 256$\mu\textrm{g}$/$m\ell$의 높은 농도에서 세포의 증식이 약간 저해되었지만 대조군에 비해 유의적인 감소는 보이지 않았다. NO 분비에 미치는 영향을 조사해 본 결과 20$\mu\textrm{g}$/$m\ell$ 농도의 백삼성분을 투여했을 때 NO분비가 대조군에 비해 유의적으로 증가하였다. Reverse transcription polymerase chain reaction (RT-PCR)을 이용하여 백삼성분에 의한NOS 유전자 발현의 정도를 조사한 결과 20$\mu\textrm{g}$/$m\ell$ 농도에서 nos gene발현이 대조군보다 증가되었다

• 식물병연구
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• 제25권4호
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• pp.233-236
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• 2019

2017년, 감귤류에 Citrus vein enation virus (CVEV)의 발생보고가 있었다. 식물방역법상 관리급 병원체인 CVEV에 대한 검역조치를 위해 감귤의 주요 재배지인 제주도를 포함한 4개도 29개 시군에서 온주밀감, 유자, 만숙성 감귤류(한라봉, 천혜향, 레드향, 황금향) 재배과원 등 203개소에서 시료를 수집하고 reverse transcription polymerase chain reaction (RT-PCR) 방법과 시퀀싱을 통해 진단하였다. 진단결과, 양성반응을 보인 시료를 대상으로 외피단백질 유전자 부위를 분석하기 위해 GenBank에 등록된 분리주들과의 염기서열과 아미노산 서열을 비교한 결과, 98% 이상의 매우 높은 상동성을 확인하였다. 전체 시료에 대하여 RT-PCR 진단 결과, 136개 과원(67%)에서 CVEV가 검출되었으며, 유자 과원의 85.4%, 온주밀감 과원의 77.8%가 CVEV에 감염된 것으로 파악됐다. 또한, 감귤의 주요 재배지인 제주도에서 90.6%의 과원이 CVEV에 감염된 것으로 확인하였다. 기주별·지역별 발생실태 조사를 토대로 국내 감귤류에 CVEV가 만연되어 있음을 확인하였다. 본 연구를 바탕으로, CVEV의 발생실태 조사와 검역병원체 여부를 검토하여 2018년 5월 CVEV를 비검역 병원체로 변경하였다.

• 대한화장품학회지
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• 제43권1호
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• pp.19-26
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• 2017

본 연구에서는 채진목의 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 RAW 264.7 세포에 대한 채진목 70% 에탄올 추출물의 효과를 살펴보았다. 채진목 70% 에탄올 추출물의 대식세포에서의 세포 독성 측정을 MTT assay를 수행하였다. 세포 독성을 측정한 결과, $1,000{\mu}g/mL$의 농도에서 96%의 세포 생존율을 나타내었다. 항염증 활성을 효과적으로 검증하기 위해, LPS로 유도된 대식세포 내 NO 생산을 억제하는 효과를 Griess의 방법으로 조사하였다. 그 결과, 채진목 70% 에탄올 추출물에서 NO의 생성이 농도 의존적으로 저해되었음을 확인하였다. 채진목 70% 에탄올 추출물을 western blot을 이용하여 단백질 발현을 측정한 결과 처리한 세포군에서 농도가 증가함에 따라 iNOS와 COX-2의 단백질 발현양이 감소하여 최고 농도인 $500{\mu}g/mL$에서 각각 84.3%, 56.2%의 발현 억제를 보여주었다. Reverse transcription-polymerase chain reaction (RT-PCR)을 통하여 mRNA 발현양을 측정한 결과 iNOS와 COX-2의 mRNA 발현양이 감소하여 $500{\mu}g/mL$ 농도에서 89.8%, 84.9%로 나타내었다. 이러한 결과로 보아 채진목 70% 에탄올 추출물의 항염증에 관한 그 효능을 확인할 수 있었고, 따라서 채진목 70% 에탄올 추출물이 천연 항염증 소재로써 가능성이 있다고 판단된다.

• 한방재활의학과학회지
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• 제24권2호
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• pp.1-13
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• 2014

Objectives The purpose of this study was to investigate the effects of Cheunggihwadamhwan (CGHDH) extract on lowering lipid, antioxidation and production of proinflammatory cytokines in rats fed on high fat diet. Methods 40 Male Sprague-Dawley rats were fed on high fat diet for 8 weeks and 32 rats (above 400 g) were randomly divided into 4 groups (8 mice in each group) : control group, 100 mg/Kg CGHDH group, 200 mg/Kg CGHDH group, 300 mg/Kg CGHDH group. We fed a control group of rats a basal diet and administered normal saline(100 mg/kg, 1 time/1 day) for 4 weeks. And We fed each experimental group of rats basal diet and administered an extract of Cheunggihwadamhwan extracts (100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid in plasma and liver, concentration of proinflammatory cytokines, antioxidative activity and gene expression. The gene expression level was investigated by the way of reverse transcription-polymerase chain reaction (RT-PCR). Results 1. Concentration of plasma FFA, plasma TG, plasma total cholesterol and plasma LDL-cholesterol showed a significant decrement in Cheunggihwadamhwan groups. However, concentration of plasma HDL-cholesterol showed a significant increment in 200, 300 mg/kg Cheunggihwadamhwan groups. 2. Concentration of liver total cholesterol and liver TG showed a significant decrement in Cheunggihwadamhwan groups. 3. Concentration of plasma TBARS showed a significant decrement in all Cheunggihwadamhwan groups. Concentration of liver TBARS showed a significant decrement in 200, 300 mg/kg Cheunggihwadamhwan groups. Concentration of liver GSH-Px, SOD and CAT showed a tendency to decrease in all Cheunggihwadamhwan groups. 4. Concentration of plasma IL-$1{\beta}$, plasma IL-6, TNF-$\alpha$ and NO, showed a tendency to decrease in all Cheunggihwadamhwan groups. Concentration of plasma IL-10 showed a tendency to increase in all Cheunggihwadamhwan groups. 5. In the analysis of reverse transcription-polymerase chain reaction (RT-PCR), the gene expression of Apo-B and Apo-E in the Cheunggihwadamhwan groups showed a low expression than that of control group. The ratio of Apo-B expression per $\beta$-actin expression in the showed a significant decrement in all Cheunggihwadamhwan groups. The ratio of Apo-E expression per $\beta$-actin expression in the showed a significant decrement in 300 mg/kg Cheunggihwadamhwan groups. Conclusions According to this study, the extract of Cheunggihwadamhwan showed a positive effect of lowering lipid, antioxidation and a control of producing proinflammatory cytokines.

• Asian Pacific Journal of Cancer Prevention
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• 제15권18호
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• pp.7825-7829
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• 2014

Objective: To investigate the effect of high expression of XAF1 in vivo or in vitro on lung cancer cell growth and apoptosis. Methods: 1. The A549 human lung cancer cell line was transfected with Ad5/F35 - XAF1, or Ad5/F35 - Null at the same multiplicity of infection (MOI); (hereinafter referred to as transient transfected cell strain); XAF1 gene mRNA and protein expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively. 2. Methyl thiazolyl tetrazolium (MTT) and annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis before and after infection of Ad5/F35 - XAF1 with Western blotting for apoptosis related proteins, caspase 3, caspase - 8 and PARP. 3. After the XAF1 gene was transfected into lung cancer A549 cells by lentiviral vectors, and selected by screening with Blasticidin, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were applied to detect mRNA and protein expression, to establish a line with a stable high expression of XAF1 (hereinafter referred to as stable expression cell strain). Twenty nude mice were randomly divided into groups A and B, 10 in each group: A549/XAF1 stable expression cell strain was subcutaneously injected in group A, and A549/Ctrl stable cell line stable expression cell strain in group B (control group), to observe transplanted tumor growth in nude mice. Results: The mRNA and protein expression of XAF1 in A549 cells transfected by Ad5/F35 - XAF1 was significantly higher than in the control group. XAF1 mediated by adenovirus vector demonstrated a dose dependent inhibition of lung cancer cell proliferation and induction of apoptosis. This was accompanied by cleavage of caspase -3, -8, -9 and PARP, suggesting activation of intrinsic or extrinsic apoptotic pathways. A cell strain of lung cancer highly expressing XAF1 was established, and this demonstrated delayed tumor growth after transplantation in vivo. Conclusion: Adenovirus mediated XAF1 gene expression could inhibit proliferation and induce apoptosis in lung cancer cells in vitro; highly stable expression of XAF1 could also significantly inhibit the growth of transplanted tumors in nude mouse, with no obvious adverse reactions observed. Therefore, the XAF1 gene could become a new target for lung cancer treatment.

• Tuberculosis and Respiratory Diseases
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• 제82권4호
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• pp.328-334
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• 2019

Background: Although the frequency of respiratory viral infection in patients with pulmonary acute respiratory distress syndrome (ARDS) is not uncommon, clinical significance of the condition remains to be further elucidated. The purpose of this study was to compare characteristics and outcomes of patients with pulmonary ARDS infected with influenza and other respiratory viruses. Methods: Clinical data of patients with pulmonary ARDS infected with respiratory viruses January 2014-June 2018 were reviewed. Respiratory viral infection was identified by multiplex reverse transcription-polymerase chain reaction (RT-PCR). Results: Among 126 patients who underwent multiplex RT-PCR, respiratory viral infection was identified in 46% (58/126): 28 patients with influenza and 30 patients with other respiratory viruses. There was no significant difference in baseline and clinical characteristics between patients with influenza and those with other respiratory viruses. The use of extracorporeal membrane oxygenation (ECMO) was more frequent in patients with influenza than in those with other respiratory viruses (32.1% vs 3.3%, p=0.006). Co-bacterial pathogens were more frequently isolated from respiratory samples of patients with pulmonary ARDS infected with influenza virus than those with other respiratory viruses. (53.6% vs 26.7%, p=0.036). There were no significant differences regarding clinical outcomes. In multivariate analysis, acute physiology and chronic health evaluation II was associated with 30-mortality (odds ratio, 1.158; 95% confidence interval, 1.022-1.312; p=0.022). Conclusion: Respiratory viral infection was not uncommon in patients with pulmonary ARDS. Influenza virus was most commonly identified and was associated with more co-bacterial infection and ECMO therapy.

• 식물병연구
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• 제27권2호
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• pp.84-90
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• 2021

2019년 6월 충남 논산의 당귀 재배 농가에서 원형반점, 괴사반점, 황화, 모자이크 등의 증상을 보이는 당귀잎을 채집하였고, 이들 시료에 대한 바이러스 감염 여부를 확인하기 위하여 전자현미경 검경과 감염우려가 있는 바이러스 3종(cucumber mosaic virus, broad bean wilt virus 2, tomato spotted wilt virus [TSWV])에 대해 reverse transcription polymerase chain reaction 진단을 수행한 결과 TSWV에 대해 양성반응을 보였다. 당귀 TSWV의 생물학적 특성을 구명하기 위해 순수분리하여 5과 28종의 지표식물과 일당귀에 즙액접종하여 기주범위와 병원성을 확인하였다. TSWV의 3 segment (L, M, S)의 전체 염기서열을 결정하고 기존에 보고된 TSWV 분리주와 계통분석을 한 결과, 당귀 TSWV 분리주(NS-AG-28)는 논산지역의 머위 분리주(TSWV-NS-BB20)와 가장 유사한 것으로 나타났다. TSWV는 넓은 기주범위를 가지고 있고 특히 고추, 토마토 등 가지과 작물에 큰 피해를 주고 있기 때문에, 당귀가 TSWV의 중간기주로서 작용하지 않도록 주의하고, 당귀의 바이러스병 피해를 예방하기 위하여 지속적인 모니터링이 요구된다.

• 한국가축번식학회지
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• 제26권4호
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• pp.329-338
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• 2002

This study was carried out to investigate the developmental rates of embryo reconstructed with different cell type and to estimate correlation of transcriptional level of octamer-binding transcription factor 4 (Oct4) and fibroblast growth factor 4 (FCF4) gene on peri-implantation stage embryos. Donor cells were transferred into perivitelline space of enucleated oocytes. The karyoplast-cytoplast couplets were accom- plished by cell to cell fusion and activated with ionomycin and 6-dimethylaminopurine. Reconstructed embryos were co-cultured with bovine oviduct epithelial cells in CR 1 aa medium. There is no difference in blastocyst formation rate following nuclear transfer UT) with fetal fibroblast cell (16/50; 32.0%), cumulus cell (16/49; 32.6%) and ear cell (17/52; 32.6%). The expression level of Oct4 and FCF4 in peri-implantation bovine embryo derived from in vitro fertilization (IVF) and NT were determined by reverse-transcription polymerase chain reaction (RT-PCR) technique. In peri-implantation of IVF result in a transient increased of FCF4 paralleled by an increased expression of Oct4. However, Oct4 gene was highly expressed in hatching blastocysts derived from NT compared to IVF. Also, FGF4 expression level in hatching blastocysts and outgrowth stage derived from NT was lower than that of IVF. In conclusion, it is suggested that the different transcription patterns observed in nuclear transfer embryos may lead to a lower rate of embryo development, implantation and pregnancy.