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.
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.
A nineteen years old male patient showed a cystic lesion in left maxillary canine to premolar area (#23-#25). This lesion was asymptomatic, and found during his routine radiological check in local clinic. In the radiological observation the cystic lesion showed round radiolucent image containing many calcified bodies which were usually small but irregular in shape, expanding tumorously and resulted in the displacement of canine and second premolar in the absence of first premolar. The lesion was surgically enucleated, and a cystic fibrous tissue containing abnormal teeth was removed and examined pathologically. With the histological observation of tumorous odontogenic epithelium including many ghost cells, which were closely associated with abortive teeth, the lesion was finally diagnosed as CCOT associated with complex odontoma. The ghost cells of CCOT was strongly positive for β-catenin, GADD45, and LC3, and slightly positive for MMP-9, while they were rarely positive for BCL2, Wnt1, HSP-70, and p38. Therefore, it was presumed that the ghost cells of CCOT might undergo dormant cell state through altered cytodifferentiation stimulated by severe growth arrest, DNA damage signaling, and abundant autophage formation.
최근 조직공학 기술의 발달로 재해와 질병으로 인한 손상된 조직과 장기의 대체연구들 이 진행되고 있다. 장기 대체 연구의 핵심 요소 중 하나는 재건된 조직이나 장기가 혈관 망을 형성하여 host tissue로부터 양분과 산소의 전달이다. 본 연구는 조직공학 기법을 이용하여 장기 재건에 필수적인 혈관 재건을 위해 혈관을 구성하는 주세포인 endothelial cell을 체외에서 배양하는 것이다. Endothelial cell(EC)배양을 위해서는 세포지지체인 세 포외 기질(External cellular metrics, ECM)을 필요로 하기 때문에 ECM중에 대표적인 collagen과 gelatin을 사용하여 지지체에 따른 체외배양능을 비교하였다. 실험 동물로는 돼지 대동맥을 채취하여, 대동맥 속에 collagenase type I을 주입하고, 혈관의 입·출구를 봉합한 상태로 10분 간 37℃에서 처리하였다. 관류된 용액은 10% FBS가 함유된 기본배 양액(EGM-2 media)을 사용하여 2번 수세한 후 회수된 세포를 각각의 ECM이 처리된 dish위에서 배양 하였다. EC세포인지를 확인하기 위해서 EC표지 인자인 CD31과 vWF 항체의 발현을 flow cytometry로 확인 하였고, 회수된 세포에서 두 단백질이 모두 발현 되었다. ECM에 따른 EC의 세포 형태를 비교하였을 때 형태학적 차이는 없었다. Basement Membrane Extract위에서 calcein-AM으로 염색된 EC는 ECM의 종류와 상관 없이 2-6시간 사이에 Tube Formation을 보였다. 또한 endothelial cell의 표지 마크인 CD31, Flk1, vWF의 mRNA 발현양과 IHC에 의한 단백질 발현을 조사한 결과 collagen 지지체 위에서 배양된 endothelial cells에서 발현양이 더 높았다. 결론적으로 두 가지 ECM에서 모두 성공적으로 endothelial cell의 배양이 가능하지만 collagen위에서 배양된 endothelial cell이 더 우수한 maintenance능력을 가짐을 확인 할 수 있었다.
The aim of this study was to compare the appearance rate of vaginal cytology during estrous cycle in small pet bitches. A characteristic features of vaginal cytology during the estrous cycle were the high proportion of superficial cell in proestrus, anuclear cell in estrus, small intermediate cell in diestrus, and small intermediate cell in anestrus, respectively. There were no statistically significant differences of appearance rate of vaginal cytology among small pet bitches in the each phase of estrous cycle. These results indicated that the vaginal cytology was useful method for estimating estrous cycle and optimal breeding time in small pet bitches.
이산화염소는 살충효과를 지니며, 이는 이 물질이 발생시키는 활성산소에 기인된다. 살충효과를 주는 주요 원인으로 이산화염소의 세포독 성에 주목하고 있다. 본 연구는 이산화염소가 유발하는 세포독성이 활성산소에 기인한 아폽토시스 유발로 가설을 세우고 이를 검증하였다. 화랑 곡나방(Plodia interpunctella) 유충에 이산화염소를 주입한 결과 전체혈구수의 뚜렷한 감소를 보였고, 이후 처리 유충은 사망하였다. 아폽토시스 세포치사과정을 규명하기 위해 TUNEL (terminal deoxynucleotidyl transferase nick end translation) 분석법을 적용하였다. 곤충 세포주의 하나인 Sf9 세포에 서로 다른 이산화염소를 처리하고 TUNEL 분석법으로 관찰한 결과 처리 농도에 비례하여 아폽토시스 비율이 증가하였다. 다 음으로 서로 다른 농도의 이산화염소를 화랑곡나방 유충에 주입하고 혈구 세포를 TUNEL 분석법으로 관찰한 결과 이산화염소는 처리 농도에 비례하여 아폽토시스 유발을 나타냈다. 그러나 항산화제인 비타민 E를 이산화염소와 함께 처리하면 비타민 E의 농도에 비례하여 이산화염소의 아폽토시스 유발을 억제하고 이에 따라 살충률도 감소하였다. 이러한 결과는 이산화염소에 기인한 세포독성은 활성산소에 기인한 아폽토시스 유 발로 이뤄졌다는 것을 제시하고 있다.
본 연구는 흰점박이꽃무지(Protaetia brevitarsis seulensis (Colbe) (Cetoniidae, Coleoptera) 유충의 혈림프에 존재하는 혈구세포들의 형태 학적 특성분석을 위하여 수행하였다. 흰점박이꽃무지 유충 혈강 내에는 과립혈구세포, 세포질혈구세포, 편도혈구세포, 구상적혈구세포, 전혈구 세포, 지방혈구세포 총 여섯 종류의 혈구세포들이 관찰 되었다. 그 중 과립혈구세포는 핵, 미토콘드리아, 골지체를 포함하여 잘 발달된 세포소기관들이 관찰 되었고 외래물질 침입시 면역학적 식균작용을 수행하는 것으로 밝혀졌다. 특히, 과립혈구세포의 세포질에는 잘 발달된 리소좀(<1 ㎛ 직경)들이 세포막 주변으로 분포되어 존재하고 있음을 알 수 있었다. 식균된 외래물질은 다양한 크기의 리소좀들과 서로 합쳐지면서 외래물질을 제거하는 것으로 판단된다. 그 외 다섯 종의 혈구세포들은 외래물질 침입시 면역학적 활성화와 관계가 없는 것으로 관찰되었다.
Nucleotide metabolism in endothelium is variable between different species. Recent studies demonstrated that this variability could contribute coagulation dysfunction, even though organs of the alpha 1,3-galactosyltransferase gene knockout pig were transplanted into the primate. CD73 (ecto-5'-nucelotidase) is an enzyme at cell surface catalyzing the hydrolysis of adenosine triphosphate to adenosine, which plays role on a substance for anti-inflammatory and anti-coagulant. Thus, overexpression of CD73 in endothelial cells of the pig is considered as an approach to reduce coagulopathy. In this study, we constructed a human CD73 expression vector under control of porcine Icam2 promoter (pIcam2-hCD73), which is expressed specifically at endothelial cells, and of CMV promoter as a control (CMV-CD73). First, we transfected the CMV-CD73 vector into HEK293 cells, and then confirmed CD73 expression at cell surface by flow cytometry analysis. Next, we transfected the pIcma2-CD73 and CMV-CD73 vectors into primary porcine fibroblasts and endothelial cells. Consequence was that the pIcma2-CD73 vector was expressed only at the porcine endothelial cells, meaning that the pIcam2 promoter lead to endothelial cell-specific expression of CD73 in vitro. Finally, we nucleofected the pIcam2-hCD73 vector into passage 3 fibroblasts, and enforced hygromycin selection of 400mg/ml. We were able to obtain forty three colonies harboring pIcam2-CD73 to provide donor cells for transgenic cloned porcine production.