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사람 HPRT 유전자 발현벡터 제조와 생쥐 Sp2/0 myeloma 세포주에서의 발현
고창보,김용만,김영진,백상기 충남대학교 기초과학연구소 1997 忠南科學硏究誌 Vol.24 No.1
To construct expression vector for the human HPRT gene, pRSVneo plasmid conferring resistance to neomycin-kanamycin(Tn5) antibiotics and pHPT 31 containing human HPRT cDNA were used to subclone human HPRT cDNA. For construction of recombinant pRSVneo carrying human HPRT, pRSVneo-HPRT, the human HPRT cDNA fragment from pHPT31 plasmid was inserted into polyadenylation site (BamHI site) behind the neo gene fragment of pRSVneo plasmid. For the other expression vector, pRSV-HPRT, the neo gene and small portion of the untranslated 5′franking region were removed from the pRSVneo-HPRT. The HPRT gene of the two vectors were constructed in right orientation. Each recombinant vector was introduced into cultured HPRT-deficient mouse cell line, Sp2/0 by calcium mediated DNA transfection and the transfected cells were selected under HAT selection condition. The HPRT activity of the lysates from the selected cell was higher than that of the lysates from spleen cells of mouse. The HPRT activity of pRSVneo-HPRT vector was higher than that of pRSV-HPRT one. To find out whether the HPRT activity of the transfected cells selected under HAT medium was expressed by transfected vectors or by spontaneous mutations, it was cultured to reselect in G418 medium. It showed that the nature of the HPRT activity was resulted from the transfected vectors.
血府逐瘀湯이 大動脈 平滑節 細胞에서 NO 生成에 미치는 影響
한종민,고창보,박창민,정명수,박길래,이기남 대한한의학회 2002 대한한의학회지 Vol.23 No.2
Effects of Hyeolbuchukeo-tang (Xiefuzhuyutang in chinese) on the NO Production in Aortic Vascular Smooth Muscle Cells
The Emerging Role of Natural Killer Cells in Innate and Adaptive Immunity
김은미,명평근,조대호,최인표,강형식,고창보 대한면역학회 2004 Immune Network Vol.4 No.4
In the early host defense system, effector function of natural killer (NK) cells results in natural killing against target cells such as microbe-infected, malignant, and certain allogenic cells without prior stimulation. NK cell cytotoxicity is selectively regulated by homeostatic prevalence between a repertoire of both activating and inhibitory receptors, and the discrimination of untransformed cells is achieved by recognition of major histocompatibility complex (MHC) class I alleles through inhibitory signals. Although it is well known that the bipotential T/NK progenitors are derived from the common precusor, functional mechanisms in terms of the development of NK cells remain to be further investigated. NK cells are mainly involved in innate immunity, but recent studies have been reported that they also play a critical role in adaptive immune responses through interaction with dendritic cells (DC). This interaction will provide effector functions and development of NK cells, and elucidation of its precise mechanism may lead to therapeutic strategies for effective treatment of several immune diseases. (Immune Network 2004;4(4):205-215)
원발성 폐암에서 혈장 과립구 자극인자의 암표지자로서의 역할과 의의
송정섭 ( Jung Sub Song ),김소영 ( So Young Kim ),조향정 ( Hyang Jeong Jo ),이강규 ( Kang Kyoo Lee ),신정현 ( Jeong Hyun Shin ),신성남 ( Seong Nam Shin ),김동 ( Dong Kim ),박성훈 ( Seong Hoon Park ),이영진 ( Young Jin Lee ),고창보 대한결핵 및 호흡기학회 2009 Tuberculosis and Respiratory Diseases Vol.66 No.6
연구배경: 폐암은 진단 당시에 완치할 수 있는 확률이 적어 예후가 불량한 종양으로 알려져 있어, 폐암 진행을 예측할 수 있는 암 표지자(tumor marker)의 발굴이 필요한 실정이다. 그러나 폐암에서 아직까지 특이적인 항원이 없고 현재까지 알려진 많은 종양관련 항원들의 민감도가 떨어지기 때문에 보편화되지 못하고 있다. 본 연구에서는 원발성 폐암 환자에서 혈장 G-CSF를 측정하고 암의 진행 및 예후와 관련이 있는지 알아보고자 하였다. 방법: 원발성 폐암으로 진단된 100명 환자와 건강 검진에서 이상 소견이 없는 127명 정상인을 대상으로 하였다. 결과: 정상인에서 혈장 G-CSF 농도는 12.2±3.6 pg/mL (mean±SD), 폐암환자에서는 46.0±38.0 pg/mL였다(p<0.001). 비소세포폐암에서 G-CSF 농도는 유의하게 소세포폐암보다 높았으며(p<0.05), 비소세포 폐암중 대세포 폐암이 가장 높았고, 편평세포암, 선암, 세기관지폐포암 순이었다. G-CSF 농도는 국소형보다는 진행형 비소세포폐암에서 증가하는 경향을 보였다. 또한 타 장기로의 전이가 있을 때 유의하게 증가하였으며(p<0.05), 다발성 전이에서는 뇌, 부신, 골 전이 순으로 혈청 G-CSF 농도가 증가하는 경향이었다. 결론: 혈장 G-CSF 농도는 폐암이 진행한 경우, 특히 타 장기로의 전이가 있을 때 유의하게 증가하였다. 그러므로 진행형 폐암의 추적관찰에 이용할 수 있으리라 사료된다. Background: Biomarkers for cancer have several potential clinical uses, including the following: early cancer detection, monitoring for recurrence prognostication, and risk stratification. However, no biomarker has been shown to have adequate sensitivity and specificity. Many investigators have tried to validate biomarkers for the early detection and recurrence of lung cancer. To evaluate plasma G-CSF as such a biomarker, protein levels were measured and were found to correlate with the clinicopathological features of primary lung tumors. Methods: Between December 2006 and May 2008, 100 patients with histologically-validated primary lung cancer were enrolled into this study. To serve as controls, 127 healthy volunteers were enrolled into this study. Plasma G-CSF levels were measured in lung cancer patients using the sandwich ELISA system (R & D inc.) prior to treatment. Results: The mean plasma G-CSF levels were 12.2±0.3 pg/mL and 46.0±3.8 pg/mL (mean±SE) in the normal and in the cancer groups, respectively. In addition, plasma G-CSF levels were higher in patients with early lung cancer than in healthy volunteers (p<.001). Plasma G-CSF levels were higher in patients who were under 65 years old or smokers. Within the cancer group, plasma G-CSF levels were higher in patients with non small cell lung cancer than in patients with small cell lung cancer (p<.05). Overall, plasma G-CSF levels were shown to increase dependent upon the type of lung cancer diagnsosed. In the order from highest to lowest, the levels of plasma G-CSF tended to decrease in the following order: large cell carcinoma, squamous cell carcinoma, adenocarcinoma, and bronchioloalveolar carcinoma. Plasma G-CSF levels tended to be higher in patients with advanced TNM stage than in localized TNM stage (I, II<III, IV). Increased levels were also seen in patients with distant metastasis in the order of bone, adrenal gland involvement. Conclusion: Plasma G-CSF level were significantly increased in patients with lung cancer, and in especially advanced TNM stage. These results suggest that plasma G-CSF can be used to support the diagnostic process of lung cancer staging and as an indicator of metastasis.