5-fluorouracil (5-FU) is a pyrimidine analog which can work as antineoplastic antimetabolite by blocking thymidylate synthetase conversion of deoxyuridylic acid to thymidylic acid in DNA synthesis. This study is aimed to know the anticancer effect of 5-FU on the expressions of important signaling proteins in KB cells through immunoprecipitation high performance liquid chromatography (IP-HPLC). KB cells were treated with 5 μM 5-FU and cultured for 12, 24, 48, 72, and 96 hours, and followed by IP-HPLC analysis using 32 antisera. 5-FU suppressed the proliferation of KB cells by decreases in the expressions of proliferation-related proteins, Ki-67, PCNA, CDK4, and MPM2 to 82.6%, 92.4%, 95.2%, and 95.9%, respectively, but increases of antiproliferation-related proteins, p16 and p21 to 106.7% and 125.5%, respectively, during 96 hours of experiment. This proliferation reduction was also negatively regulated by cMyc/MAX/MAD network signaling. The cellular protection and survival were consistently arrested by 5-FU treatment in KB cells. The expressions of NFkB, MDR, p-mTOR, and TNFα were decreased to 95.1%, 92.8%, 93.4%, and 90.3% in 48-72 hours, respectively, while cellular stress was increased by upregulation of p38 to 111.3% in 48 hours. And the expressions of pAKT1/2/3, hTERT, and AMPK were also decreased to 93.3%, 97.4%, and 89.3% in 24-48 hours, respectively, while the cellular transformation might be undergone by upregulation of TGF-β1 to 117% until 96 hours. Particularly, 5-FU treatment greatly induced the cellular apoptosis in KB cells by increased expressions of PARP, cPARP, caspase 9, c-caspase 9, caspase 8, and caspase 3 in the lack of p53/BAX and FASL/FAS signaling. The expressions of PARP and c-PARP were increased maximum to 119.2% in 24 hours, and followed by increases of caspase 9, c-caspase 9, caspase 8, and caspase 3 to 111.2%, 125.9%, 108.6%, and 116.3% in 72-96 hours. Therefore, it is presumed that 5-FU induced cellular apoptosis in KB cells may be derived from the overexpression of PARP due to the increased DNA defect caused by 5-FU, which can lead to ATP depletion and subsequent cellular apoptosis.

To gain insights into the role of purinergic receptors in human dental pulp cells (hDPCs) differentiation, we characterized the expression and functional activity of P2Y1 receptors and investigated the effects of ADP on the proliferation and differentiation of this pulp stem-like cell population. Our data showed that ADP did not induce cell proliferation to expose the various ADP concentrations for 72 hours, but the proliferative capacity of hDPCs was inhibited at higher ATP concentrations (100 μM). Using RT-PCR analysis, we found that ADP induced several P2Y receptors including P2Y1 as well as odontoblastic differentiation genes, dentin matrix protein 1 (DMP1) and dentin sialophosphoprotein (DSPP) in a dose-dependent manner. The effects of ADP on the expression of DMP-1 and DSPP mRNA were prevented by the P2Y1 antagonist MRS2179. The extracellular matrix calcium deposits were clearly observed in ADP-treated hDPCs by alizarin red S staining. Quantitative measurement of mineralization induced by ADP was significantly inhibited in MRS2179-treated hDPCs. These results may provide new insights into the molecular regulation of the differentiation of hDPCs.

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.

본 연구는 숲에서 치유를 경험한 사람들의 체험수기 115편을 질적 분석하여, 숲 치유로 나타나는 핵심 심리현상과 치유의 과정을 추적하기 위하여 수행되었다. 숲 치유 과정에서 경험하는 핵심 심리현상은 <행복감> <편안함> <깨달음> 의 순이었으며, 숲 치유의 과정은 <정서적 변화>를 먼저 경험한 후에 <인지적 변화>와 <행동의 변화>로 이끌어지는 경향을 나타내었다. 숲에서의 치유단계는 시간의 흐름에 따라 <자극단계> <수용단계> <정화단계> <통찰단계> <채움 단계> <변화단계>의 6단계 과정을 거쳤으며, 이러한 과정은 <자연과의 교감> <자신과의 교감> <세상과의 소통>이라 는 기제를 통해 치유가 진전되는 것으로 확인되었다. 기존의 연구는 주로 집단프로그램에 의한 비교 연구와 체험 후의 일시적 상태의 측정으로 결과를 도출하여 효과의 지속성이나 심리적 변화 과정을 검증할 수 없는 한계가 존재하였 다. 본 연구는 기존의 양적 연구의 한계를 보완하여 개인적 심리현상과 치유과정 등을 통합적으로 이해할 수 있는 방법을 제안하였으며, 연구 결과가 산림치유의 메커니즘을 밝히고 산림치유 프로그램 개발이나 실행에 기초자료로 활용될 수 있을 것으로 기대한다.

In order to know the characteristic roles of salivary protein complex (SPC) the gel-filtration chromatography was performed using the unstimulated and the stimulated whole saliva separately. The first and second dominant SPC peaks were fractionated and analyzed by immunoprecipitation HPLC (IP-HPLC) using antibodies against the essential salivary proteins including α-amylase, mucin-1, proline rich proteins (PRPs), histatin, cystatin, LL-37, lysozyme, lactoferrin, -defensin-1, -2, -3, IgA, transglutaminase 4 (TGase 4), mucocidin, α1-antitrypsin, cathepsin G. In the gel-filtration chromatography the stimulated whole saliva showed much reduced amount of SPCs than the unstimulated whole saliva, but the proportional patterns of both whole saliva were almost similar each other. Through IP-HPLC analysis both of the first and second dominant SPCs were variably positive for the essential salivary proteins, however, α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G were predominant in the first dominant SPC, while cystatin, lactoferrin, β-defensin-1, -2,-3, IgA, mucocidin, TGase 4, and α1-antitrypsin were predominant in the second dominant SPC. And more, the α1-antitrypsin and cathepsin G which were mostly derived from gingival crevicular fluid were also consistently found in the SPCs. These data may suggest that the first dominant SPC, rich in α-amylase, mucin-1, PRPs, lysozyme, and cathepsin G, may play a role in food digestion, protein degradation, and mucosa lubrication, while the second dominant SPC, rich in cystatin, lactoferrin, β-defensin-1, -2, -3, mucocidin, IgA, TGase 4, and α1-antitrypsin, may play a role in the mucosa protection and antimicrobial defense.

Taurine is an abundant amino acid in many animals, including humans. Relatively large amounts of taurine are found in leukocytes, heart, muscles, retinas, kidneys, bones, and liver. Taurine has antioxidant effects; it reacts with hydrogen peroxide to prevent oxidation of the cell membrane. Taurine enhances the effects of anticancer drugs, while also reducing side effects, and taurolidine, a taurine derivative, has been shown to exhibit anti-cancer effects without notable side effects in several types of cancer. Taurine aids in cholesterol metabolism by increasing the rate of synthesis of bile acids, and, thus, reduces triglyceride levels. In addition, taurine is involved in the growth and differentiation of nerve cells and is associated with some neurological disorders. Taurine aids in bone formation and prevents bone dissolution. Moreover, taurine prevents liver damage from a variety of drugs and, thus, protects the liver. Taurine is involved in the development and function of the retina and lens. It also has anti-atherosclerotic and anti-thrombotic effects that protect against cardiovascular disease. Taurine may have additional physiological functions, and warrants further investigation.

본 논문은 여성치유에 관한 진 아크터버그의 역사적 연구에 기반하여 영적 치유자로서의 여성의 역할에 대하여 재조명하고 있다. 가부장제 이전엔 언제나 여성이 치유역할을 담당했다. 치유는 본래 성스러운 영적 영역에 속했고, 여성만이 생사의 비밀을 아는 자였기 때문에, 아주 오래 전에는 문화의 중심자인 여성이 영적 치유사의 역할을 담당했었다. 궁극적인 치유는 치유사와 치유받는 자의 상호성과 더불어, 몸과 마음이 본래 둘이 아니고, 개체와 우주 역시 둘이 아니라는 영적 인식을 필요로 한다. 그러나 가부장제 하에서 여성의 치유역할은 뒷전으로 물러나고 기껏해야 보조역할로 축소되었지만, 고대문화의 흔적을 보전하고 있는 흑인여성문화에서는 여전히 여성이 영적 치유사 역할을 하고 있다. 흑인여성소설가들, 특히 뱀버러의 󰡔소금 먹는 사람들󰡕은 여성치유사가 전통적인 여성적 방식으로 치유를 하고 있음을 구체적으로 예증해주고 있다. 특히 이 소설은 소설의 시간적 배경이 되는 1970년대 화두가 되었던 새로운 휴머니즘, 새로운 영성주의가 대표하는 통합성을 흑인영성의 치유와 구원의 대안으로 제시하고 있다.

동물의 장기를 인간에게 이식하게 되면 초급성거부반응(Hyperacute rejection, HAR)이 일어난다. 초급성거부반응은 면역계의 구성요소 중 보체(complement)에 의해 일어나는 거부반응으로 돼지의 혈관세포 표면에 있는 Galα(1,3)Gal 당분자에 인간의 항체가 즉각 반응하기 때문에 일어나며, α1,3-galactosyltransferase(α1,3-GT) 유전자는 돼지 혈관세포 표면의 Galα(1,3)Gal 당분자 생성에 관여한다. 따라서 인간에게 돼지의 장기를 이식하기 위해서는 α1,3-galactosyltransferase 유전자를 제거하는 것이 필요한 것으로 알려져 있다. 본 연구실의 이전 연구에서, 시카고 미니돼지 귀체세포에서 상동 재조합(Homologous recombination)을 통해 α1,3-galactosyltransferase 유전자가 제거된 체세포를 개발한 바 있으며, 이 체세포를 통하여 α1,3-GT 유전자가 제거된 돼지도 생산된 바 있다. 본 연구에서는, human serum 처리 시 돼지 세포를 보호해 준다고 보고되고 있는 human complement regulator인 human Decay-accelerating factor(hDAF)와 human α1,2-fucosyltransferase(hHT)유전자를 α1,3-GT 유전자 위치에 gene targeting하여 동시에 hDAF와 hHT가 발현하는 체세포를 개발하였다. Knock-in vector는 hDAF와 hHT 두 유전자가 발현할 수 있도록 IRES로 연결하였으며, α1,3-GT 유전자의 start codon을 이용하여 발현할 수 있도록 구축하였다. 구축한 vector는 electroporation을 통해 미니돼지 체세포에 도입하였으며, PCR 결과, α1,3-GT 유전자 위치에서 상동 재조합이 일어났음을 확인하였다. Positivenegative 선별 방법을 통해 얻은 gene targeting 된 체세포는 RT-PCR에 의해 hDAF와 hHT 유전자의 발현이 확인되었으며, 대조군(NIH minipig)에 비해 α1,3-GT 유전자의 발현이 감소하였다. 또한 이들 세포에 100% human complement serum을 처리하였을 때 knock-in 세포가 대조군에 비해 30% 정도 더 높은 생존율을 보였다. 따라서 개발된 체세포는 이종간 장기이식을 위한 돼지 생산과 함께 이를 이용한 이종간의 장기 이식 시 초급성 거부반응을 억제하는 데 사용될 수 있을 것으로 생각된다.

Malignant gliomas and glioblastomas are the most common type of primary brain tumors. The treatment of malignant glioma involves surgery, radiation, and chemotherapy. These therapies have not been successful in curing malignant glioma and typically associated with dismal prognosis. Therefore, we can investigate thymidine kinase activity and cytotoxic effect after transfer suicide gene in U-251 glioma cells. We assessed expression patterns of green fluorescence protein(GFP) after infected with adenovirus in U-251 cells. After infection of HSV-tk in U-251 cells, we observed thymidine kinase activity with [3H]-penciclovir and cytotoxic effect by treated with ganciclovir. We could observe that expression level of GFP was increased according to infected concentrations in U-251 cells. GFP was not expressed in 1moi and 10moi, and slightly expressed in 30moi. Expression level of GFP was largely increased in 50moi and almost cells expressed GFP in 100moi. GFP expression has shown clear image in 100moi compared with other concentrations. We also investigated thymidine kinase activity using [3H]-penciclovir after infection of suicide gene HSV-tk into U-251 cells. Thymidine kinase activity increased in 10moi concentration compared with empty adenovirus infection. We could find that thymidine kinase activity was elevated proportional to HSV-tk infection amount in 30moi and 50moi. For evaluation of cellular cytotoxic effect of HSV-tk, we treated ganciclovir to U-251 cells and assessed cytotoxicity by using MTT assay. We could identifiy that cytotoicity appeared in very low concentration of HSV-tk compared with cancer cells originated with other organs. Cytotoxic effect was shown about 15% of U-251 cells of total cells in 5moi. By infection 10moi of HSV-tk, cytotoxic effect was intensively increased and about 60% of U-251 cells became extinct. About 70% cells exhibited cytotoxic effect in 30moi and more than 80% cells also appeared cytotoxic effect by infection of HSV-tk in 50moi, 100moi, and 200moi. Therefore, we could confirm to gene expression in U-251 cells was increased proportional to infected gene concentrations. Also we could find that thymidine kinase activity elevated with according to infected concentration and cellular cytotoxic effect was shown in very low concentration and higher cytotoxic effect also appeared by infection of suicide gene HSV-tk into U-251 glioma cells. These results suggest that gene therapy with suicide gene will be successful in curing brain tumors containing malignant glioma and glioblastoma.

Terfenadine (TFN) was a second generation histamine receptor antagonist. Although several studies have reported the regulatory effect of H1-histamine receptor antagonists in human cancer cell lines, its effect in oral cancer remains unclear. In this study, we focused on addressing the anti-cancer activity of TFN in human oral cancer cell lines. The anti-cancer activities of TFN were performed by tryphan blue exclusion assay, 4'-6-diamidino-2-phenylindole (DAPI) staining, live/dead assay and Western blot analysis. TFN induced a significant reduction of the growth in three different human oral cancer cell lines (MC3, HSC4 and Ca9.22). TFN markedly induced apoptosis through DNA damage and increase in cytotoxicity. It also accumulated cleaved PARP and caspase 3. This process was due to cleavage of caspase 8 and Bid protein. The results from this study strongly demonstrated that the cleavages of caspase 8 and Bid are required for the apoptotic activity of TFN in human oral cancer cells. Taken together, these findings suggest TFN as a potent anticancer drug candidate for the treatment of oral cancer.

Salivary proteins include numerous functional proteins which play important roles not only for the food-intake but also for the protective and defensive mechanisms. In the present study the compositions of salivary proteins were analyzed by different methods, including electrophoresis and high performance liquid chromatography (HPLC). In hydrophobic protein HPLC analysis the parotid saliva gradually produced macromolecular complexes when agitated in refrigerator until 30 minutes. These salivary protein complexes were digested by neuraminidase, and then migrated more rapidly in native tris glycine gel than the control. Therefore, it was assumed that the glycosylated proteins of parotid saliva became gradually aggregated to form salivary protein complexes similar to those of whole saliva. The salivary protein complexes were easily degenerated in different experimental buffers, i.e., SDS buffer, tris glycine buffer, methanol, etc., and resulted non-specific patterns in electrophoresis and HPLC. Therefore, it was presumed that the salivary protein complexes was made by the hydrophobic interaction as well as electrostatic attraction between salivary proteins. These data indicated that to know the real pattern of salivary protein complexes in vivo the whole saliva should be analyzed by HPLC using non-adhering column with isoelectric buffer. Consequently, the whole saliva was analyzed by HPLC using reverse phase SuperoseTM column with 20 mM potassium phosphate buffer, and two prominent peaks of salivary protein complexes were consistently found in every people. These salivary protein complex peaks were relatively stable up to 6 hours after saliva collection when the whole saliva was kept in refrigerator during experiment. Therefore, it is suggested that the salivary protein complex patterns are characteristic macromolecular structures of whole saliva, which are also applicable as a diagnostic point in different saliva-associated diseases

Caffeic acid phenethyl ester (CAPE), a component of propolis, was reported to possess anti-inflammatory, anti-bacterial, anti-viral, and anti-tumor activities. Our aim was to investigate the effect of CAPE on apoptosis in cultured human mucoepidermoid carcinoma (MEC) cell line, MC-3. Apoptotic effects of CAPE were measured by cell viability assays, Western blotting, 4’-6-diamidino-2-phenylindole (DAPI) staining and Live/Dead assay. The result of cell viability assay showed that CAPE displayed a strong growth-inhibitory effect in a concentration-dependent manner against MC-3 cells. Consumption of CAPE resulted in pronounced increase in the cleavage of caspase-3 and PARP, induced nuclear condensation and fragmentation and clearly increased the number of dead cells in MC-3 cells. CAPE also caused the increase in truncated Bid (t-Bid) and the cleavage of caspase-8 and this phenomenon was regulated by death receptor 5 (DR5). In addition, Phosphorylation of AKT and ERK were downregulated by CAPE. Taken together, these results suggest that CAPE is a potent apoptosis-inducing agent in MC-3 cells.

한국, 중국, 일본 3국 공통의 「상용한자 800자 선정」초안이 2013년 일본 홋카이도에서 발표되었다. 이것은 한중일 한자 문 명의 역사를 향해 새로운 한걸음을 내디딘 것을 보여주고 있 다. 동아시아 공유 가치를 확산하고 3국이 미래의 교류를 보다 활성화 하자는 인식에서였다. 본고는 한국에서 본연의 한글 교 육 정책에 따른 한자 교육을 통해서 보다 좋은 유용한 방향성 을 고찰하였다. 먼저 이를 위해 세계 공통어문, 공통 문자권과 한문, 한자 문화권의 역사적 배경과 한글과 한자의 역사적 관 계에 대해 정리하였다. 또한 일본의 한자 문제나 중국 한자와 비교하고, 한자・한문을 둘러싼 현재의 문제와 해결 방법을 고 찰하였다. 특히 일본의 한자 시책 및 인쇄 기술에 따른 국제 표준 규격에 대한 선행 연구 사례를 조사 분석하여 문자는 누 구를 위해 있는지 문자의 표현 기술 및 한자 서체 디자인 및 인쇄 기술 등에 관하여 재정리을 하였다. 그 결과 한국의 문자 교육에 있어서 관련 연구자들과 기초 자료로 공유가 기대된다.

본 연구의 목적은 한우사료 내 아마씨앗의 급여가 등심 내 오메가 6와 3 지방산 비율(n-6/n-3) 감소효과 및 n-6/n-3 균형 한우고기를 섭취한 사람의 혈액 중 LDL-C 감소효과를 조사하는 것이었다. 단계별로 거세한우 20마리를 이용하여 대조군과 아마씨앗을 함유하는 n-3 처리구로 각각 10마리씩 나누어 완전임의 배치하였다. n-3 사료군은 대조군과 비교할 때 혈액 및 등심 내 n-6/n-3가 4:1이하로 감소하였으며 단일불포화지방산으로써 올레인산은 52.79%까지 증가하였다. 임상실험자의 70% 이상에서 나타난 균형 한우고기를 섭취한 그룹의 중성지방, 총콜레스테롤 및 LDL-C은 각각 25.35, 5.22, 17.59% 감소하였고 수입 쇠고기는 9.05, 8.21, 21.70% 증가하였으나 일반한우는 큰 차이가 나타나지 않았다. 균형 한우고기를 섭취한 그룹의 HDL-C는 6.07% 증가하였으나 수입 쇠고기와 일반한우는 각각 14.46, 11.46% 감소하였다. 혈당은 균형 한우고기와 일반 한우고기가 각각 6.42, 11.82% 감소하였으나 수입 쇠고기는 15.19% 증가하였다.

J. M. 싱의 시학 읽어내기: 「아란 섬」과 「바다를 타는 사람들」에 나타난 자연과 시 우리말 요약: J. M. 싱은 아마 지금까지 아일랜드에서 태어난 가장 위대한 극작가들 중의 하나이다. 그러나 그는 자주 많은 이들에 의해 폄하(조이스 포함하여)되고 있지 만 예이츠는 당대에 모든 극작가들 중 최고로 여기는 것 같다. 본 논문은 싱이 아란 을 방문하기 전에 이미 잘 준비된 위대한 극을 쓸 수 있었다고 주장하며, 「아란 섬」을 증거로 제시하는데, 그가 나중에 희곡을 쓸 때 사용하는 관찰, 느낌, 생각으로 가 득 찬 아주 시적인 위대한 저술임을 증명한다

본 연구에서는 자발적인 웃음과 인위적인 웃음을 변별하는 데 있어서 일반 사람들의 정확도와 컴퓨터를 이용한 분류 알고리즘의 정확도를 비교하였다. 실험참가자들은 단일 영상 판단 과제와 쌍비교 판단과제를 수행하였다. 단일 영상판단 과제는 웃음 영상을 한 장씩 제시하면서 이 영상의 웃음이 자발적인 것인지 인위적인 것인지를 판단하는 것이었으며, 쌍비교 판단과제는 동일한 사람에게서 얻은 두 종류의 웃음 영상을 동시에 제시하면서 자발적인 웃음 영상이 어떤 것인지 판단하는 것이었다. 분류 알고리즘의 정확도를 산출하기 위하여 웃음 영상 각각에서 8 종류의 얼굴 특성치들을 추출하였다. 약 50%의 영상을 사용하여 단계적 선형판별분석을 수행하였으며, 여기서 산출된 판별함수를 이용하여 나머지 영상을 분류하였다. 단일 영상에 대한 판단결과, 단계적 선형판별분석의 정확도가 사람들의 정확도보다 높았다. 쌍비교에 대한 판단결과도 단계적 선형판별분석의 정확도가 사람들의 정확도보다 높았다. 20명의 실험참가자 중 선형판별분석의 정확도를 넘어서는 사람은 없었다. 판별분석에 중요하게 사용된 얼굴 특성치는 눈머리의 각도로, 눈을 가늘게 뜬 정도를 나타낸다. Ekman의 FACS에 따르면, 이 특성치는 AU 6에 해당한다. 사람들의 정확도가 낮은 이유는 두 종류의 웃음을 구별할 때, 눈에 관한 정보를 충분히 사용하지 않았기 때문으로 추론되었다.

Recently, extensive research has been performed in the field of orthopedic medicine to develop cell-based therapies for the restoration of injured bone tissue. But there has been rarely reported about rehabilitaton of oral and maxillofacial bone defect using self-derived osteoblasts. Normal human osteoblast cell(NHost) was previously established into marrow-derived human mesenchymal stem cells for their capacity to proliferate and differentiate into osteoblasts under various culture conditions. The purpose of this study was to examine proliferation and differentiation of NHosts effected by growth factors with ALP activity and RT-PCR. After NHosts were cultured under basal and osteogenic medium at 37℃ and 5% CO2, they were analyzed by ALP activity and RT-PCR. BMP-2 under osteogenic medium decreased growth rate of NHosts compared to under osteogenic medium. BMP-2 under osteogenic medium induced osteoblastic differentiation in NHosts by increased ALP activity. The differentiating capacity of NHosts under osteogenic medium showed that NHosts expressed higher mRNA expression levels of OSX and OCN, while that of RUNX2 decreased after BMP-2 treatment. It suggested that NHosts having characteristics of osteoprecursor cells might be more advanced in their osteogenesis development by BMP-2, making NHosts an interesting biological tool for treatment of skeletal defects and diseases of oral and maxillofacial bone.