• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6557-6562
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• 2013

Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.

• Asian Pacific Journal of Cancer Prevention
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• 제14권7호
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• pp.4215-4221
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• 2013

Purposes: Lung cancer is prevalent worldwide and improvements in timely and effective diagnosis are need. Pentraxin-3 as a novel serum marker for lung cancer (LC) has not been validated in large cohort studies. The aim of the study was to assess its clinical value in diagnosis and prognosis. Methods: We analyzed serum PTX-3 levels in a total of 1,605 patients with LC, benign lung diseases and healthy controls, as well as 493 non-lung cancer patients including 12 different types of cancers. Preoperative and postoperative data were further assessed in patients undergoing LC resection. The diagnostic performance of PTX-3 for LC and early-stage LC was assessed using receiver operating characteristics (ROC) by comparing with serum carcinoembryonic antigen (CEA), cytokeratin 19 fragments (CYFRA 21-1). Results: Levels of PTX-3 in serum were significantly higher in patients with LC than all controls. ROC curves showed the optimum diagnostic cutoff was 8.03ng/mL (AUC 0.823, [95%CI 0.789-0.856], sensitivity 72.8%, and specificity 77.3% in the test cohort; 0.802, [95%CI 0.762-0.843], sensitivity 69.7%, and specificity 76.4% in the validate cohort). Similar diagnostic performance of PTX-3 was observed for early-stage LC. PTX-3 decreased following surgical resection of LC and increased with tumor recurrence. Significantly elevated PTX-3 levels were also seen in patients with non-lung cancers. Conclusions: The present data revealed that PTX-3 was significantly increased in both tissue and serum samples in LC patients. PTX-3 is a valuable biomarker for LC and improved identification of patients with LC and early-stage LC from those with non-malignant lung diseases.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5731-5734
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• 2013

Background: The object of this study was to examine whether a new protocol including step-sectioning and immunohistochemistry (IHC) staining of axillary sentinel nodes (SN) would lead to detection of more metastases in patients with breast cancer. Materials and Methods: Sixty-nine tumor free sentinel lymph nodes were examined. Step frozen sectioning was performed on formalin fixed SN and stained both by hematoxylin and eosin (H and E) and cytokeratin markers using IHC. Any tumoral cell in IHC stained slides were considered as a positive result. Metastases up to 0.2 mm were considered as isolated tumor cells and 0.2 up to 2 mm as micrometastasis. Results: Mean age of the patients was $48.7{\pm}12.2$ years. Step sectioning of the SN revealed 11 involved by metastasis which was statistically significant (p<0.001). Furthermore, 15 (21.7%) of the patients revealed positive results in IHC staining for pan-CK marker and this was also statistically significant (p=0.001). Ten patients had tumoral involvement in lymph nodes harvested from axillary dissection and 4 out of 15 lymph nodes with positive result for CK marker were isolated tumor cells. However, 4 of 10 patients with tumor positive lymph nodes in axillary dissection were negative for CK marker and in contrast 6 of the pan-CK positive SN were in patients with tumor-free axillary lymph nodes. Conclusions: Both IHC and step sectioning improve the detection rate of metastases. Considering the similar power of these two methods, we recommend using either IHC staining or step sectioning for better evaluation of harvested SNs.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권4호
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• pp.322-330
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• 2007

Backgrounds: To overcome limited amount of autogenous mucosa for the reconstruction of various mucosal defect including oral mucosal defect, tissue engineered mucosa has been recently introduced. However, introduced conventional technique of tissue engineered mucosa still have serious pitfalls such as long fabrication time, fragility of the reconstructed mucosa, and complexity of the technique. Aim of the study: To examine whether the complex of preconfluent autologous keratinocytes and autologous PRP(Platelet rich plasma) can reconstruct oral mucosa on the muscular flap with easier and faster way compared to conventional mucosal tissue engineering technique. Materials and methods: One day before the operation, oral mucosa(3mm in diameter) were taken and treated for extraction of oral keratinocytes according to the routine manner. The day of operation, oral keratinocytes were prepared in the laboratory and then moved to the operating theater. Autologous PRP was also prepared and then mixed with oral keratinocytes just before grafting on the prepared muscular flap. After keratinocyte-PRP complex was seated, then a sterilized rubber sheet was placed on the graft and the elevated skin flap was replaced and sutured. Biopsies were proceeded at 3, 5, 7, 14 and 21 days. Tissue samples were evaluated clinically, histologically, and immunohistochemically. Results: All of the oral keratinocyte-PRP complexes were successfully grafted on the recipient sites(100%). On 3 days after the operation, 1-2 continuous epithelial layer and many inflammatory cells were observed. On 5 days after the operation, increase of layers of keratinocyte was observed with less inflammatory response. Thickness of the layers was gradually increased from 7 to 21 days after the operation. Cytokeratin confirms epithelium in every specimen. Conclusions: Preconfluent graft of autogenous oral keratinocytes mixed with autogenous PRP have successfully reconstructed myo-mucosal flap. This technique could be a useful alternative for oral mucosal reconstruction in the near future.

• 대한소아치과학회지
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• 제30권1호
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• pp.10-14
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• 2003

근유종은 소아에 주로 발생하는 다발성 종양으로 처음에 기술되었지만, 단독으로 발생하는 경우가 더 많고, 어느 연령층에서나 발생한다. 성인과 소아형 모두 비슷한 임상적 특성과 병리조직학적 특징을 갖는다. 이 증례는 9세 남아의 하악에 경계가 명확한 단방성의 방사선 투과성 병소로, 병리조직학적으로 방추형 세포들이 다발을 이루며 증식하며, 일부에는 혈관주피종과 유사한 부위가 관찰되었다. 면역조직화학적으로 종양세포는 vimentin과 smooth muscle actin에 양성, S-100, desmin, cytokeratin AE1/3에 음성이었다. 이 증례에서와 같이 소아에서 단방성 방사선 투과상의 고형성 종괴로 나타나는 경우 근섬유종을 감별진단에 포함시켜야 한다고 생각된다. 환자는 수술 1년이 경과한 현재까지 재발의 소견은 없지만 재발한 경우도 보고되어 있어 철저한 추적 관찰이 필요하며, 현재 #43, 44 치아의 맹출을 유도하는 중에 있다.

• 한국발생생물학회지:발생과생식
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• 제14권3호
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• pp.155-162
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• 2010

The trophectoderm is one of the earliest cell types to differentiate in the forming placenta. It is an important for the initial implantation and placentation during pregnancy. Trophoblast stem cells (TBSCs) develop from the blastocyst and are maintained by signals emanating from the inner cell mass. However, several limitations including rarity and difficulty in isolation of trophoblast stem cells derived from blastocyst still exist. To establish a model for trophoblast differentiation, we isolated TBSCs from human term placenta ($\geq$38 weeks) and characterized. Cell cycle was analyzed by measuring DNA content by FACS analysis and phenotype of TBSCs was characterized by RT-PCR and FACS analysis. TBSCs have expressed various markers such as self-renewal markers (Nanog, Sox2), three germ layer markers (hNF68, alpha-cardiac actin, hAFP), trophoblast specific markers (CDX-2, CK7, HLA-G), and TERT gene. In FACS analysis, TBSCs isolated from term placenta showed that the majority of cells expressed CD13, CD44, CD90, CD95, CD105, HLA-ABC, cytokeratin 7, and HLA-G. Testing for CD31, CD34, CD45, CD71, vimentin and HLA-DR were negative. TBSCs were shown to decrease the growth rate when cultured in conditioned medium without FGF4/heparin as well as the morphology was changed to a characteristic giant cell with a large cytoplasm and nucleus. In invasion assay, TBSCs isolated from term placenta showed invasion activities in in vivo using nude mice and in vitro Matrigel system. Taken together, these results support that an isolation potential of TBSCs from term placenta as well as a good source for understanding of the infertility mechanism.

• Tuberculosis and Respiratory Diseases
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• 제47권4호
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• pp.478-487
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• 1999

연구배경 : 전자현미경적 관찰에 의해 진단된 흉막의 악성중피종과 전이성선암 환자를 대상으로 하여 7종의 단일클론항체를 이용한 면역조직화학염색을 시행하고 이들의 발현양상을 비교하여 두 질환을 감별할 수 있는 가장 효과적인 염색방법을 알아보고자 하였다. 방 법 : 면역조직화학염색법은 Fisher사(Fisher scientific)의 1-hour immunohistochemistry방법을 이용하였다. 종양조직은 포르말린으로 고정된 파라핀 조직을 이용하여, $3{\mu}m$ 두께로 잘라서, $85^{\circ}C$에서 가열시킨 후 파라핀을 제거하고, 희석액과 단클론항체를 $45^{\circ}C$에서 15분간 반응시켰다. 각 단계마다 automation buffer로 실온에서 세척하였고, avidin-biotin-peroxidase system을 이용한 immunoperoxidase method로 염색하였다. 조직절편은 $45^{\circ}C$에서 10분간 색소원반응을 시행하였고, 대조염색은 hematoxylin으로 실시하였다. 조직 절편은 최소한 각기 다른 두 구역에서 세포질 또는 세포막이 분명하게 염색될 때 양성으로 판정하였다. 결 과 : 7종의 단일클론항체중 CK, EMA, vimentin, S-100 단백과 Leu-M1은 악성중피종과 전이성선암에서 발현율의 차이는 없었으나, B72-3은 전이성선암에서만 발현되었고, CEA는 전이성선암 전례와 악성중피종 42.9%에서 발현되어서 민감도는 높았으나 특이도는 낮았다. 결 론 : B72-3을 이용한 면역조직화학염색은 악성중피종과 전이성선암의 감별진단에 가장 유용한 방법으로 시사되었으며, 기존에 사용되고 있는 CEA와 B72-3을 함께 이용한다면 악성중피종과 전이성선암에 대한 감별진단력을 높일 수 있을 것으로 사료된다.

• Tuberculosis and Respiratory Diseases
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• 제42권6호
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• pp.846-854
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• 1995

연구배경: Cyfra 21-1은 상피종양세포의 세포질에 존재하는 cytokeratin 19의 분절로서 상피종양세포의 파괴시 혈중내로 유리되므로 그 혈중 농도를 측정하여 종양표지자로 이용할 수 있는 것으로 알려져 있다. 이에 저자들은 폐암, 폐결핵, 기타폐질환 및 정상대조군 환자들의 혈청내 Cyfra 21-1, SCC 항원 및 CEA의 농도를 측정하여 폐암의 종양표지자로서 Cyfra 21-1과 SCC 항원 및 CEA의 진단적 효용성을 비교 관찰하고자 하였다. 또한 편평상피세포암에서 Cyfra 21-1과 평상펴셰포암의 특이 종양표지자로 알려진 SCC 항원과의 진단적 민감도와 특이도의 차이를 비교하고 그 병기 진행에 따른 Cyfra 21-1의 혈중농도의 증가 여부를 관찰하고자 하였다. 방법: 1992년 12월부터 1993년 6월까지 서울중앙병원에 입원하여 조직생검으로 초진단된 원발성 폐암 79예(편평상피세포암 41예, 선암 18예, 기타의 미분화 비소세포양 14예, 소세포암 6예)와 폐결핵 32예, 기타폐질환 23예, 정상대조군 23예를 대상으로 하였다. Cyfra 21-1과 ELSA2-CEA를 사용하였고, SCC 항원은 방사계수측정 kit인 ABBOTT SCC RIABEAD를 사용하였다. 결과: 1) Cyfra 21-1의 혈중농도는폐암군이 평균({\pm}표준편차) $18.38{\pm}3.65\;ng/mL$로서 비교군 $1.16{\pm}0.53\;ng/mL$보다 유의하게 높았다(p<0.0001). SCC 항원은 폐암군에서 $3.53{\pm}6.06\;ng/mL$로서 비교군 $1.19{\pm}0.5\;ng/mL$보다 유의하게 높았다(p<0.01). CEA는 폐암군에서 $35.03{\pm}13.9\;ng/mL$로서 비교군 $2.89{\pm}1.01\;ng/mL$ 보다 유의하게 높았다(p<0.0001). 2) 폐암군내에서는 Cyfra 21-1의 혈중농도가 편평상 피세포암군에서 $31.52{\pm}40.13\;ng/mL$로서 선암군 $2.41{\pm}1.34\;ng/mL$(p<0.0001) 및 소세포암군 $2.15{\pm}2.05\;ng/mL$(p=0.007) 보다 유의하게 높았다. SCC 항원의 혈중농도는 편평상피세포암군에서 $5.1{\pm}7.68\;ng/mL$로서 선암군 $1.36{\pm}0.69\;ng/mL$(p=0.009) 및 소세포암군 $1.1{\pm}0.24\;ng/mL$(p=0.024)보다 유의하게 높았다. 3) 편평상피세포암군에서 폐암의 병기 진행에 따른 Cyfra 21-1의 혈중농도의 증가는 없었다. 4) Cyfra 21-1의 진단양성 기준치를 3.3 ng/mL로 하였을때 편평상피세포암의 민감도가 83%로 선암의 22%, 소세포암의 17%보다 높게 산출되었다. SCC 항원의 민감도가 편평상피세포암에서 39%, 선암에서 11%, 소세포암에서 0% 이었다. CEA의 민감도가 편평상피세포암에서 20%, 선암에서 39%, 소세포암에서 33%이었다. 5) ROC 곡선 분석상 폐암의 진단에서 Cyfra 21-1의 민감도와 특이도가 SCC 항원 및 CEA 보다 우수한 것으로 나타났다. 결론: Cyfra 21-1은 폐임에서 SCC 항원 및 CEA에 비하여 민감도 및 특이도가 높은 종양표지자이며, 특히 편평상피세포암에서 그 민감도와 특이도가 높아 편평상피세포암의 특이 종양표지자로 알려진 SCC 항원보다 우수한 종양표지자로 사료되었다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제27권6호
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• pp.510-522
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• 2005

To assess the clinical applicability of bio-artificial mucosa which was made with autologous oral keratinocytes and human acellular dermal matrix, the formation of basement membrane and stratification of oral keratinocytes were evaluated. Six New Zealand white rabbits (around 2kg in weight) were anesthetized and its buccal mucosa was harvested (1.0 $\times$ 0.5cm size). Oral keratinicytes were extracted and cultured primarily with the feeder layer of pretreated NIH J2 3T3 fibroblast. These confluent cells were innoculated on the human acellular dermal matrix and cultured in multiple layer by air-rafting method. After 3, 5, 7, 10, 14 days of culture, each cultured bio-artificial mucosa was investigated the number of epthelial layer of by H&E stain and toluidine blue stain. The immuhohistochemical methods were used to evaluate the cell division capacity, the formation of basement membrane, and it's property of specific cells (PCNA, cytokeratin 14, laminin). Transmission electromicroscopy was used for the attachment between cells and matrix with the number of hemidesmosome. In result, the numbers of layer of stratified growth of oral keratinocyte cultured on the human acellular dermal matrix and the number of hemidesomal attachment between epithelial cells and human acellular dermal matrix were similar to the layers of normal oral mucosa after 10 days of culture. The cell division rate, basement membrane formation and proliferation rate increased as culture period increased. With these results, bio-artificial mucosa with autologous oral epithelial cells cultured on the acellular dermal matrix had clinically adaptable properties after 10 days' culture and this new bio-artificial mucosa model with relatively short culture time can be expected clinical applicability.

• 핵의학기술
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• 제17권2호
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• pp.101-106
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• 2013

Purpose: As UBC IRMA is being tested, patients out of the reference value are reacting within the value again a few days later the urine collection tested, which causes the reliability of the test to decrease as a result. In this study, we can assume that the physiological changes in the factors would affect the results. The purpose of the study is to find out whether hematuria and proteinuria in the sample as well as the interval time (3 hours or more recommended) have affected the results. As a result, we could discover the changes in factors and increase the reliability of the test. Materials and Methods: 468 people (female: 249, male: 219) who came for the check-up were presented herein for medical examination from 2013.3.15 to 2013.1.2. Some people out of 468 who have reacted onto the reference value were divided into group low titer zone, ow-middle titer zone, and middle-high titer zone and tested for hematuria and proteinuria. During that period, 48 outpatients were asked to fill in a questionnaire regarding the urination interval time. The reagents used were (IDL Biotech AB, Sweden) and UBC IRMA. Results: Of the patients that are formed in the reference value of ($0.1-34.0{\mu}g/L$) turn out to be 52.7 years average age in their low concentration, ($mean{\pm}SD$) of the value of $0.10{\pm}0.02{\mu}g/L$. Among 80 people (50.8%, female: 49.2%), 16 patients (20%) have shown reaction to microscopic hematuria and 10 patients (12.5%) responded to proteinuria. In the average low concentration under 52.5 years of average age, 43 people (53%) have shown reaction to microscopic hematuria and 21 people (26.3%) are proteinuric patients out of 80 patients (male: 50.8%, female: 51.3%). In the middle high concentration of $11.8{\pm}4.82{\mu}g/L$ under the average age 51.7 years, 35 patients (53%) have responded to the microscopic hematuria and proteinuric patients are 26 people (39.3%) out of 66 people (men: 44%, women: 56%). In addition, in the concentration of $51.7{\pm}43.5{\mu}g/L$, some patients who get out of the reference value are observed as the average age of 52.0. 11 patients (78.6%) out of 14 (male: 35.7%, female: 64.3%) react to the microscopic hematuria. There show 6 people (42.8%) who turn out to be as proteinuric patients. As for the interval time, $1.67{\pm}3.71{\mu}g/L$ was the average value among 48 patients (female: 45.8%, male: 54.2%). Conclusion: We cannot see if proteinuria and hematuria directly affect abnormal results of inspection of 8,18 cytokeratin; however, we can find out that they statistically have an influence on highly generating UBC among several mechanisms. Also, although urination interval time was various every 15 minutes, we it does not affect these results.