본 연구는 우엉 뿌리 추출물의 항산화 활성 및 피부에 대한 안전성을 평가하기 위하여 총 폴 리페놀 함량, 총 플라보노이드 함량, DPPH radical 소거 활성을 통하여 항산화 활성을 살펴보고, B16F10 melanoma 세포에 대한 세포 독성 및 자외선 A에 대한 피부 세포 보호 효과를 확인하였다. 또 한 화장품 소재로서의 활용을 검증하기 위하여 1차 피부 첩포 테스트를 실시하였다. 본 실험 결과 우엉 뿌리 추출물의 함량이 증가됨에 따라 높은 폴리페놀과 플라보노이드의 함량이 확인되었으며, DPPH radical 소거 활성을 확인하였다. B16F10세포에 대한 세포 독성을 확인한 결과 B16F10 melanoma 세포 에 대해 독성이 낮고, 자외선 A에 대해 80% 이상의 세포 보호 효과를 확인하였다. 또한 1차 피부 첩포 테스트를 통해 우엉 뿌리 추출물이 피부에 자극이 거의 없음을 확인하였다. 이러한 결과를 통하여 우엉 뿌리 추출물이 항산화 활성과 자외선에 대한 피부 보호 효과가 뛰어나고 피부 세포에 대한 독성이 낮으 며, 피부에 대한 안전성이 확인됨에 따라 화장품 소재로서의 가능성을 확인하였다.

The knock-in efficiency in the fibroblast is very important to produce transgenic domestic animal using nuclear transfer. In this research, we constructed three kinds of different knock-in vectors to study the efficiency of knock-in depending on structure of knock-in vector with different size of homologous arm on the β-casein gene locus in the somatic cells; DT-A_cEndo Knock-in vector, DT-A_tEndo Knock-in vector I, and DT-A_tEndo Knock-in vector II. The knock-in vector consists of 4.8 kb or 1.06 kb of 5’ arm region and 1.8 kb or 0.64 kb of 3’ arm region, and neomycin resistance gene(neor) as a positive selection marker gene. The cEndo Knock-in vector had 4.8 kb and 1.8 kb homologous arm. The tEndo Knock-in vector I had 1.06 kb and 0.64 kb homologous arm and tEndo Knock-in vector II had 1.06 kb and 1.8 kb homologous arm. To express endostatin gene as transgene, the F2A sequence was fused to the 5’ terminal of endostatin gene and inserted into exon 7 of the β-casein gene. The knock-in vector and TALEN were introduced into the bovine fibroblast by electroporation. The knock-in efficiencies of cEndo, tEndo I, and tEndo II vector were 4.6%, 2.2% and 4.8%, respectively. These results indicated that size of 3’ arm in the knock-in vector is important for TALEN-mediated homologous recombination in the fibroblast. In conclusion, our knock-in system may help to create transgenic dairy cattle expressing human endostatin protein via the endogenous expression system of the bovine β-casein gene in the mammary gland.

Recently, the area of marine resources has become concerned with sources for the next generation of the bio-industry. Until present, development of the marine resources has remained limited, although a large number of these resources are considered to have potential for various significant biological activities. Most marine sponges, marine algae and coral could be used to create specific compounds for survival against a harsh environment. Therefore, it was necessary that these materials needed to be elucidated with biological activities, such as like anti-inflammatory, anti-viral or anti-cancer effects for their utilization in the bio-industry. In this study, we screened extracts of marine resources for their anti-cancer effect on human colorectal cancer cells. These resources were collected at Kosrae of Micronesia on April, 2013 and extracted with methanol. Cytotoxicity of marine resources was observed. Of a total of 20 specimens, three specimens dose-dependently demonstration inhibition of cell viability. Furthermore, cells treated with these specimens for 48h were induced p53, p21, Bax and caspase-3. The results suggest that they involved p53-mediated apoptosis. Two positive specimens (1304KO-327 and 1304KO-329) were verified as the identical materials, which are Hyrtios sp. Unfortunately 1304KO-207 was not yet classified and needed to identify in the further study. There results suggested that marine resources with positive potential in anticancer effect would be good candidates as useful bio-resources.

Ameloblastic fibrosarcoma (AFS) is an extremely rare malignant odontogenic tumor characterized with benign ameloblastic cells islands and malignant mesenchymal component. While two-thirds of AFS seem to arise de novo, but one-third develops from recurrent ameloblastic fibroma (AF) or ameloblastic fibro-odontomas (AFO). Pathological distinction of malignant transformation is essential for appropriate treatment. The patient was a 28 years old man. Since the primary tumor was excised, the mass recurred 2 years later. The recurrent tumor was diagnosed as AFS. Chief complaint was pain in the right mandible. Computer tomography finding revealed multilocular intrabony lesion with radiopaque substance in the primary lesion. In the recurrent lesion cortical bone destruction was found. Microscopically, both the primary and recurrent lesions showed benign ameloblastic follicles with myxoid or highly cellular mesenchymal proliferation. The histological difference between primary and recurrent lesions were that foci of dental hard tissue composed of enamel and dentin were found only in the primary lesion, whereas nuclear pleomorphism was aggrevated in the recurrent lesion. The histological criteria determining malignancy were discussed.

Porphyromonas gingivalis is a gram-negative bacteria of rod shape, and grown in an anerobic condition. It colonizes in subgingival crevice and is known as a major pathogen causing chronic periodontitis. It possesses an invasive property and replicative potential within various cell types, presumably playing an important role in modulating biological behaviors of oral cancer. However, the pathophysiology of P. gingivalis in the malignant transformation of oral cancer has not been fully understood. In this study, we aimed to investigate molecular changes of oral squamous cell carcinoma cells induced by repetitive P. gingivalis infection that clinically resembles chronic periodontitis.

Lycorine, a natural alkaloid extracted from the Amaryllidaceae plant family, was reported to various physiological and pharmacological effects including anti-cancer activity. Nevertheless, there is no report of the anticancer effect of lycorine in oral cancer cells. The effects of lycorine on cell proliferation and apoptosis were examined through trypan blue exclusion assay, 4’-6-diamidino-2-phenylindole (DAPI) stain, Live/Dead assay, Western blot analysis and RT-PCR. Lycorine suppressed cell viability and induced apoptosis in MC3 and HSC-3 cell lines. Lycorine decreased survivin protein but did not affect its mRNA. It regulated survivin through accelerating protein degradation in a time-dependent manner although neither proteasome nor lysosome was not associated with lycorine-mediated protein degradation. Collectively, our results suggest that lycorine may be a potential therapeutic anti-cancer drug candidate for the treatment of human oral cancer.

The objective of this study was to investigate the effects of a lettuce (Lactuca sativa L.) extract on the inflammation of human umbilical vein endothelial cell (HUVEC) and blood lipid improvement in hypercholesterolemic mice fed a high cholesterol diet. The lettuce extract (100% ethanol extract) inhibited the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVEC treated with tumor necrosis factor-α (TNF-α). The lettuce extract suppressed the adhesion of THP-1 to TNF-α-treated HUVEC. The lettuce extract decreased the TNF-α-stimulated production of proinflammatory cytokine interleukin-6, interleukin-8 and chemokine monocyte chemotactic protein 1. In hypercholesterolemic mice, the lettuce extract reduced serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol level, while the lettuce extract elevated high-density lipoprotein-cholesterol level, resulting in the decrease of atherogenic index and cardiac risk factor level. These results suggested that lettuce extract can be an useful resource to show an anti-inflammatory effect and improve lipid metabolism

The purpose of this study was to evaluate the protective effect of PineXol® on H2O2-induced cell death in SK-N-MC cells, and in early stage focal ischemia rodent model. SK-N-MC cells were pre-treated with 200 μM H2O2 or various concentrations of PineXol® (10, 30, and 50 pg/mL) for 24 h, and then exposed to H2O2 for 3 h. Cell death was assessed by the CCK-8 assay, reactive oxygen species (ROS) assay, and lactate and dehydrogenase (LDH) release assay. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) expressions were also analyzed by western blotting. Focal ischemia rodent model was used as the in vivo model, and different concentrations of PineXol® (1, 10, and 100 mg/kg) were administered. One week after administration, reduction of infarct volume was analyzed by TTC staining. Cell viability of H2O2-treated SK-N-MC cells significantly increased by pre-treatment of PineXol® (p<0.05). PineXol® pre-treatment also induced significant decrease of ROS and LDH expressions. However, PineXol® did not affect the infarct volume. These results suggest that PineXol® has significant neuroprotective effect in vitro, but statistical significance was not confirmed in the in vivo focal ischemia mo

NNK (4-(methylnitrosamino)―1-(3-pyridyl)-1-butanone) is a major form of nitrosamine abundant in cigarette smoke and is a powerful carcinogen. Mercury is a major component of the amalgam that is widely used as dental filling material. Concurrent exposure to these two agents may result in their interaction and alter their carcinogenic potential. The present study used an immortalized human epithelial cell system that allows continuous exposure to potential carcinogens, in an attempt to elaborate the carcinogenic potential of mercury and NNK in humans. Cytotoxicity of mercury chloride and NNK was measured by an MTT assay. Parameters of neoplastic cellular transformation such as cell saturation density, soft-agar colony formation, and cell aggregation were analyzed to examine the carcinogenic potential of mercury chloride and NNK. The study showed that exposure to mercury chloride with NNK resulted in increased soft agar colony formation and cell aggregation. ROS generation by mercury chloride was further enhanced by treatment with NNK. The apoptosis that was observed following mercury chloride exposure was further increased upon co-treatment with NNK. The interaction between these two agents was also observed in cytokine mRNA induction. In the present study, mercury alone did not seem to pose a significant threat as a carcinogen, but it may have potential to enhance the carcinogenic potential of a known carcinogen from cigarette smoke. The present study provides valuable data regarding the evaluation of potential carcinogenic risk of mercury chloride and NNK on concurrent exposure.

Recently, the importance of inflammation in carcinogenesis has been recognized and studied extensively. As a result, a clear correlation between inflammation and carcinogenesis has been well established in some types of cancers. Despite a high prevalence of chronic periodontitis, one of the most common inflammatory diseases in the general population, there are only a few reports on the role of chronic periodontitis in oral cancer progression. In this study, we aimed to investigate genetic changes in oral cancer cells induced by repetitive Porphryomonas gingivalis infections to mimic chronic periodontitis in a clinical setting. Cells of oral squamous cell carcinoma (OSCC), the most common type of oral cancer, and P. gingivalis 381 were used for the present study. ID1 and ID3 were mRNAs of higher expression in the P. gingivalis-infected group compared to the uninfected control. These mRNAs have been regarded as important modulators participating in cancer progression. Future studies will provide an insight into the roles of the molecules we identified in oral cancer progression. Outcomes from these studies will also shed light on the significance of chronic periodontitis induced by bacterial pathogen, such as P. gingivalis, in progression of oral cancer and relevant molecular mechanisms underlying altered cancer cell behaviors.

혈관내큰B세포림프종은 악성 림프구의 혈관 내 성장을 보이는 드문 질환으로, 말초혈액 또는 혈관 외 종괴를 보이지 않는다. 이 림프종은 빠른 파종 및 공격적 성향 때문에 나쁜 예후를 가진다. 그러나 질병특유소견이 없어 진단이 어려운 실정이다. 저자들은 십이지장 위장관기질종양의 수술 검체 내에서 혈관내큰B세포림프종이 진단된 증례를 발견하였기에 문헌고찰과 함께 보고하는 바이다.

저자들은 상복부 불편감으로 발견된 바터팽대부 점막하 종양의 진단과 치료를 위해 시행한 내시경적 절제술상 신경 절세포 부신경절종과 이소성 췌장이 동시 발현된 증례를 경 험하여 문헌고찰과 함께 보고하는 바이다.

The aim of this study was to investigate change of plasminogen activators (PAs) and their inhibitors (PAIs) mRNA and protein expression level by heat stress in porcine endometrial cells. The endometrial epithelial cells were isolated from endometrial epithelium in porcine uterus and cultured in different temperature conditions (38.5 and 41.5℃) for 24 h. Expression of urokinase-type PA (uPA), tissue-type PA (tPA), PA inhibitor-1 (PAI-1) and -2 (PAI-2) mRNA in epithelial cells were analyzed using reverse transcription-PCR and protein levels were measured by immunofluorescence. In result, mRNA expression of uPA, tPA, PAI-1 and PAI-2 were decreased in 41.5℃ than 38.5℃ culture condition, however, significant differences were no detected. uPA, tPA and PAI-2 protein were mainly expressed in nucleus, whereas PAI-1 was distributed in cytoplasm and nucleus. uPA and tPA protein levels were increased by heat stress treatment and significant difference was only detected in tPA level (p<0.05). In contrast, two types of PAIs protein level were decreased in 41.5℃ cultured group compared with 38.5℃ group. In present study, tPA protein expression was upregulated by heat stress in porcine endometrial cells. This result suggest that change of tPA by heat stress may be related to blood flow into uterus and intrauterine microenvironments, and could directly and indirectly influence to reproductive performance in pigs.

목 적 : 본 연구는 수술 전 양성 및 악성 간 병변 환자에 대하여 자유호흡기법(free breathing technique)을 사용하여 b-value (50, 500, 800, 1000)를 변환시켜 조영제 주입 전·후 현성확산계수가 어떤 수치적인 변화를 나타내는지 연구하고 악성 종양의 평가 자료로 유용한지 알아보고자 하였다. 대상 및 방법 : 본 연구는 후향적 연구로써 임상윤리위원회(institutional review board, IRB)의 승인(PNUHIRB-17)을 얻어 진행하였으며 연구 기간은 2015년 11월 01일부터 2016년 01월 30일까지 부산 소재 일개 P대학교 병원을 내원하여 수술전 간 MRI 검사를 의뢰 받은 환자 38명을 대상으로 하였다. 연구에 사용된 프로토콜(protocol)은 본원에서 시행되고 있는 간 검사에 최적화된 검사기법(TR:5100 ms, TE: 79 ms, SPAIR, NEX: 6, b-value type: 50, 500, 800, 1000)으로 조영제 주입 전 축삭면 확산강조영상을 자유호흡기법으로 영상을 획득하고 PRIMOVIST 10 ml (Gd-EOB-DTPA) 주입후 20분 지연기(hepatobilliary phase)에 동일하게 반복 시행하였다. 사용된 확산강조영상은 SE single-shot EPI (echo planar imaging)을 이용하여 b-value값을 50, 500, 800, 1000 s/mm2으로 세분화하여 검사를 하였다. 정량적 평가시 Image J와 work station을 사용하여 평가 하였으며 세분화 되어있는 b-value 값에 따라 ADC map을 나타내고, 이에 대하여 병변에 관심영역을 설정하여 현성확산계수를 구하였다. 통계적 분석은 대응표본 t-Test를 사용하였으며, p < 0.05 일때 통계적으로 유의하게 평가되었다. 결 과 : 간세포암의 경우 조영제 주입 전과 후의 현성확산계수의 수치는 pre ADC (500, 800, 1000: 1.238, 1.040, 1.007 × 10-3 s/mm2), post ADC (500, 800, 1000: 1.225, 1.094, 1.002 × 10-3 s/mm2)를 나타내었다(p>0.05). 간전이암의 경우 pre ADC (500, 800, 1000: 1.450, 1.472, 1.332 × 10-3 s/mm2), post ADC (500, 800, 1000: 1.438, 1.441, 1.354 × 10-3 s/mm2)를 나타냈고(p > 0.05), 혈관종의 경우 pre ADC (500, 800, 1000: 1.591, 1.365, 1.217 × 10-3 s/mm2)와 post ADC (500, 800, 1000: 1.906, 1.614, 1.396 × 10-3 s/mm2)는 변화 있는 결과를 나타내었다(p>0.05). 결 론 : 결론적으로 양성 종양의 경우 조영제 주입 전과 후의 결과의 연관성이 크지 않았고, 규칙성이 없었다. 하지만 간세포암의 악성병변(LR-5)의 경우 조영제 주입 후가 양성 병변에 비해 통계적으로 의미 있게 낮은 현성계수의 값을 보였으며, 정성적 분석에 향상을 가져왔다. 본 연구에서 제시한 바와 같이 복부 확산강조영상의 경우 질병과 검사 순서에 따라 현성확산계수의 결과 값과 확산강조영상의 명확도가 다르게 측정 될 수 있으므로 조영 전과 후의 확산강조영상을 병행한다면 좋을 것이라고 사료되며, 조영후의 현성확산계수와 이용 가능한 검사 시퀀스를 병행한다면 양성 및 악성 질환 감별의 진단에 효율성을 가져올 것이라고 판단되었다

국내 자생종인 물오리나무(Alnus incana subsp. hirsuta(Spach) A. Lö ve & D. Lö ve)와 수우물오리(Alnus incana subsp. tchangbokii Chin S. Chang & H. Kim)는 형태학적인 차이로 식별 가능하지만, 유전학적인 차이는 아직 밝혀지지 않았다. 다만 플라보노이드 분석 자료와 DNA자료에 근거하여 수우물오리가 물오리나무의 배수체라는 가설이 제기된 바 있다. 이를 검증하기 위해 서울대학교 인근 관악산에서 물오리나무 21점, 수우물오리 24점의 잎을 채집하여 유동세포계수법(flow cytometry)을 통해배수성을 측정하였다. 그 결과 물오리나무는 2배체와 4배체가 나타난 반면, 수우물오리는 2배체만 존재하는 것으로 나타났다. 기존의 가설과 달리 수우물오리의 형태학적 특성은 배수성에서 기인하는 것이아니며, 배수성을 통해 수우물오리와 물오리나무를 동정할 수는 없다는 것을 밝혀냈다. 물오리나무의배수성에 대해서는 추가적인 연구가 필요할 것이다.

In order to perform the biological investigation of coffee extract containing different molecules, it would be necessary to develop in vitro experimental system rather than animal experiment. Although the animal experiment treated via oral intake or intravenous injection may disclose the whole systemic effect, the in vitro cell culture experiment would be more convenient to analyze direct cellular effect of coffee extract than animal experiment. Therefore, this study was aimed to develop a dialysis method for the crude coffee extract to perform the biological investigation using murine macrophage cell line, RAW 264.7. First of all, the RAW 264.7 cells treated with dialyzed coffee extract were observed, and subsequently their protein extracts were analyzed by gel filtration chromatography, thin layer chromatography, and immunoprecipitation high performance liquid chromatography (IP-HPLC). Resultantly, it was found that the low dose (20μg/mL) of dialyzed coffee extract, about 5 cups of ordinary coffee drinking for human adult, enhanced the growth of RAW 264.7 cells by increased expression of β-actin and Ki-67, and also induced the anti-inflammatory effect by decreased expression of NFkB, TNFα, and LC3 contrast to the high dose (40μg/mL) of dialyzed coffee extract. The low dose of dialyzed coffee extract produced almost no harmful effect on RAW cell culture for 12 hours, rather than it produced stimulatory effect on RAW cells by increasing the cell number and enhancing the protein expression of β-actin, Ki-67. Therefore, it was thought that the low dose of dialyzed coffee extract is applicable to cell culture experiment without difficult purification procedures of coffee elements. In addition, as the contrast cellular effect between the low and high dose of coffee extract was found in this study, it was also presumed that the low dose of coffee extract may play an important role in the inflammatory reaction of murine macrophages.

The fruit of Kochia scoparia Scharder is traditionally used as a medicinal ingredient to treat allergic skin diseases and inflammatory diseases in China, Japan and Korea. Recently, several studies reported that K. scoparia had potential for the cytotoxicity of human cancer cells. To investigate the anti-cancer effect of K. scoparia on oral cancer and to determine the specific type of cell death induced by MEKS treatment. We investigated the anti-cancer effects of K. scoparia, methanol extract (MEKS) in HSC4 human oral cancer cells. We examined the effects of MEKS on the proliferation rate, cell cycle arrest, 7-AAD-ANNEXIN V double stain, reactive oxygen species (ROS) generation and activation of apoptosis and necroptosis-associated proteins in HSC4 cells. MTT assay results demonstrated that MEKS decreased the proliferation rates of HSC4 cells in a dose-dependent manner with an IC50 value of 45.3 μg/ml. MEKS at 50 μg/ml significantly increased the sub-G1 DNA contents of HSC4 cells to 84.8%, versus untreated cells. However, the activation of apoptosis-associated proteins such as cleaved caspase 3, cleaved caspase 8, cleaved caspase 9 and cleaved Poly (ADP-ribose) polymerase (PARP) did not detect. The level of Bax protein markedly increased in MEKS-treated HSC4 cells. In addition, the cell viability of the DPQ pre-treated HSC4 cells with MEKS treatment was significantly greater than that of MEKS treated-cells. These results suggest that MEKS inhibits cell proliferation and induces necroptosis in oral cancer cells and that MEKS may have potential chemotherapeutic value for the treatment of human oral cancer.

우리나라 버섯산업은 자동화, 대량생산시스템이 구축되면서 급속하게 성장하여 2010년 생산량 20만 톤 그리 고 생산액으로 8,860억 원을 달성하였으나 2010년을 정점으로 해서 소비량이 줄어들고 수출이 둔화되어 버섯 산업 전체가 지금 현재 정체상태에 있는 상황이다. 정체된 이 버섯산업을 극복하고 새로운 버섯산업의 성장 동력을 창출하기 위해서 기존의 5대 버섯 외에 새로운 품종을 개발해서 부가가치가 높고 수출 활성화를 도모 할 수 있는 시장 맞춤형 버섯품종을 개발하였다. 육성 경위를 살펴보면 2013년에 품종출원한 아위느타리 ‘비 산1호’와 국내외 수집된 백령느타리 20균주에 대한 자실체 특성을 조사하여 2개의 우수균주를 선발하였다. 2014년 우수균주 가운데 수량성이 뛰어난 아위느타리 ASI 0629(비산1호)와 백령느타리 ASI 0663(백령20)의 단포자를 분리하여 Mon-Mon 교배법으로 교잡하였다. 2014년 200여개의 교잡주 중에서 자실체 형태는 백령 느타리를 띠면서 아위느타리의 미토콘드리아 DNA를 가진 세포질전환 종간교잡주 8계통을 선발하였다. 2015 년 생산력 검정시험을 통해 저온처리없이 백령느타리 형태를 띠는 고품질 우량계통을 선발하여 그 우수성이 인정되어 2015년 ‘설원’이라는 명칭으로 특허 출원하였다.