송악산과 사람 발자국 화석지 주변에서 측정된 절대 연대 측정 결과와 야외 지질 조사를 근거로 할 때, 하모리-송악산 지역의 층서는 하부로부터 광해악 현무암, 명명되지 않은 퇴적층, 송악산 응회암, 하모리층 및 사구층으로 구성되어 있으며, 발자국 화석이 산출되는 사계리 지역은 하부로부터 광해악 현무암, 사람 발자국 화석 산출 지층 그리고 사구층으로 구성되어 있다. 하모리층은 송악산 응회암이 형성된 후 퇴적된 지층이라는 규정과 절대 연령 측정 결과 및 현지에서 조사된 바에 의하면, 사람 발자국 화석이 산출되는 지층은 하모리-송악산 지역의 송악산 응회암층 상부에 놓이는 하모리층이 아니라 하부에 분포하는 명명되지 않은 퇴적층과 대비된다. 따라서 사람 발자국 화석의 형성 시기는 14C 측정 결과인 약 15,000년 전으로 해석하는 것이 합리적으로 생각된다.

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatory effects of LED irradiation such as promotes stimulation of wound healing have been well known, there are few reports about molecular mechanism associated with cell cycle by LED irradiation. The purpose of present study was to examine the molecular event in cell cycle of LED irradiation on primary human gingival fibroblast(hGF) in vitro. The source of light for irradiation was a continuous-wave LED emitting at a wavelength of 635nm, and manufactured that energy density was 5mW/cm2 on sample surface. The hGF were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental samples were acquired at 0 (right after irradiation), 8 and 24 hour after irradiation. To investigate the molecular mechanisms associated with cell cycle, growth phase was determined by flow cytometry and mRNA expression of cyclin A, cyclin B, cyclin D1, cyclin E, cdc2, PCNA, p18, p27, p21, and p53 were determined by real time RT-PCR. Flow cytometric analysis demonstrated the percentage of cells in the G1 and S phase were decreased, but the G2 phase increased, which showed cells irradiated by LED were transitioned from S to G2 phase. For mRNA expression, cyclin B, cdc2, PCNA and p53 were increased at 0 hour after irradiation, and most of cell cycle molecules were increased at 8 hour after irradiation. At 24 hour after irradiation, cyclin A, cyclin E, PCNA and p18 were increased. Taken together, LED irradiation induced proliferation of hGF cells through transition from S to G2 phase.

건설업계는 제조업과 특성이 달라 여러 가지 문제점이 엿보인다. 특히, 건설업 근골격계 질환에 대한 연구 및 지원은 별로 찾아볼 수가 없다. 근골격계 질환의 제조업과 건설업의 비율을 보았을 때 "2004 산업재해분석" 요양자 업종별 분포를 보면 제조업이 3,281명(79.79%)로 건설업의 63명(1.53%)보다 월등히 많은 것으로 조사되었다. 이는 미국의 사례를 보았을 때, 2007년 업종별 분포를 보면 건설업이 42,867명(26.20%), 제조업이 101,437명(36.23%)로 우리나라에 비해 건설업 근골격계 질환자가 상대적으로 많은 것으로 조사되었다. 이는 건설업에서의 근골격계 부담 작업 실태조사 등을 통하면 제도적 문제점이나, 근로자가 몰라서 질병에 이환되는 경우가 아닌가 한다. 건설업 근로자들은 건설업 특성상 근골격계 질환예방을 위한 도구를 거의 사용하지 못하고, 작업의 연속성, 지속성이 떨어지므로 부자연스럽고, 불편한 자세를 작업을 위해 수시로 반복하여 취할 수 밖에 없다. 따라서 건설업체에 중대재해를 줄이기 위한 노력 뿐 만 아니라, 열악한 건설업체의 근로자의 근골격계 질환 예방에 대한 제도적인 지원과 안전담당자 교육, 근로자에 대한 근골격계 질환 예방교육(스트레칭, 작업 자세, 동작, 근력강화, 적절한 휴식시간 부여) 등 대책이 절실히 필요하고 하겠다.

Oral squamous cell carcinoma is the 1st most common malignancy in oral and maxillofacial area. HPV 16 has been strongly linked to progression of cervical carcinoma. E6 and E7 as a small DNA virus encoding two major oncoproteins of HPV 16 can act together to produce efficient immortalization of primary human epithelial cells. Thus it is important to pursue the development of Immortalized human oral keratinocyte(IHOK) culture model which could be related to the pathogenesis of oral squamous cell carcinoma. If we establish IHOK transfected by E6/E7 genes, IHOK will be accepted as a model system for HPV-linked oral carcinogenesis. The purpose of this study were to culture primary normal human oral keratinocyte(NHOK), and to establish IHOK for studying oral carcinogenesis in the future. NHOK was primarily cultured under normal culture condition, and transformed into IHOK by transfection of E6/E7 genes. After 100 passages depend on Ca++ condition, cultured IHOK was confirmed by growth curve, cornified cell envelope measurement, TGase 1activity, mRNA detection, tumorogenecity and anchorage independence assay. After 100 passages, cultured IHOK showed most basal cell and monolayer of polyhedral cells under 0.15mM Ca++, and small area of stratification and flattened epithelial cells with irregular border under 1.2mM Ca++. The cultured IHOK showed relatively resistant growth under high calcium condition. The E6/E7 mRNA was detected in cultured IHOK by RT-PCR. During the terminal differentiation in cultured IHOK, increased insoluble cornified cell envelope formation was accompanied with induction of TGase 1 activity. But the cultured IHOK showed less CEM and TGase 1 activity than those of cultured NHOK. Cultured IHOK showed non-tumorogenecity, but slight anchorage independence. We had developed a technique to transform NHOK into IHOK by transfection of E6/E7 genes. Cultured IHOK was established as intermediate stage cell to study the pathogenesis of human oral squamous cell carcinoma.

Amino acid transporters are essential for the growth and proliferation in all living cells. Among the amino acid transporters, the system L amino acid transporters are the major nutrient transport system responsible for the Na+-independent transport of neutral amino acids including several essential amino acids. The L-type amino acid transporter 1 (LAT1) is over-expressed to support cell growth in malignant tumors. The double stranded RNA-mediated RNA interference (RNAi) analysis can be in a wide variety of eukaryotes to induce the sequence-specific inhibition of gene expression. In this study, we examined the effect of LAT1 short interfering RNA (siRNA) on cell growth using siRNA of LAT1 in the KB human oral squamous cell carcinoma. In the RT-PCR analysis and western blot analysis, the siRNA of LAT1 inhibited expressions of LAT1 mRNA and protein. The uptake of [14C]L-leucine was inhibited by siRNA of LAT1. In the MTT assay, the siRNA of LAT1 inhibited the growth of the KB cells in the time-dependent manner, indicating that the growth inhibition of KB cell by the siRNA of LAT1 is induced by the blocking of neutral amino acid transport mediated by LAT1. These results suggest that the transport of neutral amino acids including several essential amino acids into the KB human oral squamous cell carcinoma is mediated mainly by LAT1. Further, the LAT1 would be a new target for the inhibition of cancer cell growth.

Cultured normal human oral kera tinocyte(NHOK) & inunortalized human oral keratinocyte(IHOK) provide a valuable model in ce llular proliferation and differentiation after proper stimulation , And it is interesting to study these estab lished cell lines esca ping normal control on their growth and differentiation, SPRR1 is induced during t erminal differ entiation 0 1' human epiderma l kerat inocytes but is rarely in anaplastic cells of keratinocyte origin, But SPR1 expression has not yet been explained during differ entiation uf NHOK and Lransformed oral keraLinocyLes , The purpose of this study were to examine mHNA and protein expression of SPR1 in response to a known differentiation signal, calcium conc in NHOK, lHOK a nd oral SCC ce ll line(HN 4) , and to apply these results for investigating the molecular mecha nisms of tra nsformed cellular differentiation , Primary cultured NHOK, established IHOK and HN 4 cell line were cul tured in KBM bullet kit Preconfluency of NHOK as control group was used Under O, 15mM Ca++ conc(Precon, Postcon) , and 1, 2mM Ca++ conc(Pos tcon)‘ the insoluble final pellets were measured fo1' cornified cell envelope measurements, and RT- PCR for SPRR1 mRNA meas urement, and immunoblotting for SPRR1 protein measurements in tripli cate , resp ectively , The terminal different ia tion of cu ltured NHOK and IHOK was depend on calcium concentration, while HN4 cell line was not SPRR1 mRNA and protein expression of cultured NHOK showed the highest among cultured IHOK & HN 4 cell line in hi gher ca lcium condition , SPRR1 mRNA and protein expression of cultured IHOK showed higher‘ than tha t of HN 4 cell line in hjgher cacium condi tion , SPRH1 was expressed in differentiation of NHOK and IHOK t ransfected by E6/E7 genes but ra rely expressed in malignant oral keratinocytes , It suggested that SPRR1 ex pression as kera tinocyte terminal diff‘erent ia tion marker involved in cellular cornification would be differentially effected by immorta li zation and ca rcinogenic transforma tion

It is well kwon that HPV have been strongly linked to progression of or al squamous cell carcinoma‘ Effici ent im mortalization of nonnal human oral keratinocyte(NHOK) should provid further evidence for the role HPV in tumorogenes is ‘ Because IHOK(I mmortali zed human oral keratinocyte) has been considered as a moclel syst em for study ing I-!PV- linkecl oral ca rcinogenes is , it is important to pursue the differenti ati al change of IHOK cul t ure moclel during t he culture passage, The purposes of this study were to examine the cha ra r’ ct eristic clifferential changes of cul turecl immorta lizecl human ora l keratinocytes during long term passage, and to apply these results to or al carcinogenes is in the future, NI-!OK was primarily incubated at 370C and 5% C02 under KBM bullet kJt IHOK was co ntinuously cul t ured towarcl 100th passage(two times per week) , Growth curve of NI-!OK and II-!OK clepend on clùture passage was taken For examining the cha racte ri s t ic clifferential changes of II-!OK, transrnission electron microscope, 1ì'ansgluta miase activity‘ E6/E7 mRNA detect ion, a ncl tumorogenecity were done 10th II-!OK showecl sl ight polygonal flattencl cells and sometimes apoptotic cells ‘ while 100th IHOK showecl increased polygonal cell s ‘ Cultu recl 100th IHOK showed r ela tively resis tant growth to high calcium than 10th II-!OK Microvilli from 10th II-!OK was not connect ecl with each other, ancl scatte red cytokeratin fil aments of 10th II-IOK. while decreased cytokeratin filaments in cytoplasm & prominent clesmosome of 100th IHOK. During the terminal differ entiation in cultured IHOK, induction of TGase 1 activity of 10th II-!OK was higher than that of100th IHOK mRNA E6E7 expresson was cletected and unchangable in both cul tured cells There was no tumorogenecity inclucecl by both culturecl cel ls. Although late passage IHOK showecl less r esemblance to NHOK, and lower TGase 1 acti vi ty than ea rly passage IHOK, it suggested that these cells should be 110t yet fully differ entiatecl to oral squ a mous ce ll carcinoma cells

Cervical carcinoma is the 1st most common malignancy in korean females. HPV have been strongly linked to progression of cervical carcinoma. E6 and E7 as a small DNA virus encoding two major oncoproteins of HPV can act together to produce efficient immortalization of primary human epithelial cells, providing further evidence for the role of HPV in tumorogenesis. It is important to pursue the development of Immortalized human epithelial keratinocyte(IHEK) culture model which could be related to the pathogenesis between cervical and oral carcinoma. If we establish IHEK transfected by E6E7 gene, IHEK will be accepted as a model system for HPV-linked cervical carcinogenesis. The purpose of this study were to culture primarily normal human epithelial keratinocyte(NHEK), and to establish IHEK for applying these results to cervical and oral carcinogenesis in the future. The obtained results were as follows. 1. After 7-9 passages, cultured NHEK was almost senesce and disappeared, but cultured IHEK showed most basal cell and monolayer of polyhedral cells under 0.05mM Ca++, while small area of stratification and flattened epithelial cells with irregular border under 1.2mM Ca++. 2. The cultured IHEK showed relatively resistant growth to high calcium condition. 3. The mRNA E6E7 in cultured IHEK by RT-PCR was detected. 4. During the terminal defferentiation in cultured NHEK and IHEK, increase of insoluble cornified cell envelope formation was accompanied with induction of TGase 1 activity. But the cultured IHEK showed less CEM and TGase 1 activity than those of cultured NHEK. 5. Cultured IHEK showed non-tumorogenecity, but week anchorage independence. From the aboving results, we have developed technique to transform NHEK into IHEK by transfecting cells with E6E7 gene. Cultured IHEK was established as intermediate stage cell for studying the pathogenesis of human cervical carcinoma.

The study of cornified cell envelope has been used to investigate the differentiation factors and to advance oral carcinogenesis, CE of human oral keratinocytes are in wet condition as saliva containing many proteases, growth factors, and many kinds of bacteria, The analysis of CE in Immortalized human oral keratinocyte(IHOK) derived from normal human oral keratinocyte(NHOK) will be used to study the pathogenesis of oral squamous cell carcinoma, The purpose of this study was to analyze the amino acid component derived from CE of cultured NHOK and IHOK, It will be helpful to study the role of transfected E6/E7 gene in forming CE, and to examine the pathogenesis of oral squamous cell carcinoma, After primry culture of NHOK, IHOK were cultured in KBM bullet kit at 370C under 95% C02 incubator, Growth curve according to calcium concentration, cornified cell envelope measurement(CEM), and protein chemistry for amino acid component of CE were done(Mena :f::SD) , respectively. The obtained results were as follows, lHOK showed small areas of stratification, more compact, with irregular border and tightly apposed cells in 1,2 mM Ca++, Cornified cell envelope exhibited an aggregated group of empty space surrounded by the remained cell membrane, During the terminal differentiation in cultured NHOK and IHOK, insoluble cornified cell envelope formation was increased, CEM of NHOK was about 4 folds than that of lHOK under high calcium, Amino acid component of both groups showed Pro/Glu(SPR) , Gln/Glu(lnvolucrin) , and Gly(Loricrin) in descending order, From the aboving results, ít was suggested that when the terminal dífferentiation in cultured NHOK and IHOK, major amino acid component of CE in cultured lHOK was the same to that of cultured NHOK, It was thought that E6 and E7 gene should be involved in preventing the differentiation and proliferation of IHOK from making CE,

In order to obtain novel genes related to the human craniofacial development, molecular cloning and sequencing, and in situ hybridization using craniofacial tissue sections were performed and followed by protein structure simulation. Totally 231 clones were obtained from the subtracted craniofacial tissue cDNA library of human embryo. Random cloning using the non-redundant clones from the craniofacial tissue of human embryo was done and obtained 398 clones from the premade human chondrocyte cDNA library. Their partial sequence data showed that 214 clones of subtracted cDNA library of craniofacial tissue were still non-redundant in Genebank search. And 20 clones among 498 clones of premade chondrocyte cDNA library were known to be undefined genes. Through in situ hybridization screening in the craniofacial tissue sections of 10 weeks old human embryo 36 clones were found to be positive in specific tissues. Depending on the cell types of sirnilar developmental origin, the positive reactions could be divided into five groups. Among the 20 clones of undefined genes from human chondrocyte cDNA library, 7 clones showed characteristic positive reaction in human cartilage tissue by in situ hybridization. From the simulated protein structure, motif analysis and in situ hybridization studies for the 7 undefined clones, Ch89, Ch96, Ch129, Ch285 clones may function in the outer space of the cell constituting a part of matrix protein complex, and Ch276 as a transmembrane protein which might partic ipate in matrix calcification around chondrocytes. Ch153 is a kind of antirnicrobial protein also acting as an inflammation mediator, and Ch334 clone is a zinc finger protein, of which expression increases in human adult tissues We presume these novel genes from human chondrocytes may provide a new path of chondrocyte development and functions of human craniofacial tissues

약용식물내 에스트로겐성과 항-에스트로겐성을 조사하고 항암인자를 발견하기 위하여, 본연구는 에탄올추출로 제조된 9종류의 한국산 약용식물에 대하여 재조합효모와 MCF-7 사람유방암세포주를 이용하여 스크리닝하고 비교하였다. 재조합효모를 이용한 실험결과, 7종류의 약용식물에서 에스트로겐성이 나타났고, 4종류에서 안드로겐성이 나타났다. 또한 MCF-7 사람유방암세포주를 이용한 실험결과, 8종류의 추출물이 MCF-7 세포의 성장을 억제하는 것으로 확인되었으며 비스페놀 A와 동시 처치한 경우에도 유의적으로 억제하는 것으로 나타났다. 또한 Clyeyrrhiza uralensis, Cassia tora, Syringa velutina, Zingiber officinale, Malva verticillata, Panax ginseng C.A. Meyer는 식물성 에스트로겐으로서 에스트로겐에 양성인 사람유방암세포의 증식을 유의적으로 억제시티는 흥미로운 결과가 제시되었다. 따라서 이번 연구는 한국산 약용식물이 식물성 에스트로겐과 항암인자로서 이용될 수 있으며, 에스트로겐의 활성을 조사하는데 유용하게 이용될 수 있을 것으로 사료된다.

Oral cavity offers a good environment for the bacterial growth. 까le species diversity of oral bacteria has been studied for many years based on the classification of liable bacteria. In order to acquire more comprehensive insight into the baα.erial community of human oral cavity, we used molecular ecological methods to clone 16S rDNAs from human saliva. DNA was direαly extracted from saliva collection of human according to age. By 27F, 1492R primer for 16s rDNA’s amplification were enforced. Cloning was achieved from the amplified DNA using pGEM-T Easy Veαor and competent cellJM 109. 159 different base sequences were obtained from saliva samples of four different persons, chosen according to the age group. In all samples, Streptococcus sp. were the most abundant rnicrobes, followed by Prevotella sp., except in the saliva of an old person where Rothia sp. presented the second dominant group 까lis saliva sample showed another and more irnportant characteristics that there were increased species diversity, from pathogenic bacteria like Haemophilus sp. and lautropia sp. to the higher pr'φ。rtion of unculturable bacteria. The same tendency, but less clearly, was found for the saliva of adult smoker in the forties. These results suggest that environmental factors in the oral caviψ of smoker and older person makes favourable condition for the infectious bacteria.

This study focuses on some evangelism models to reach secular people. John Wesley’s “ Order of Salvation" model, James Engel’s “ Count Down" model, Agnes Liu ’ S “ Triangle" model, George Hunter's “ Target" model, and Joseph Aldrich’s “ Relationship" model are reviewed. John Wesley’s “ Order of Salvation" model is effective in understanding how secular people become Christians. Further, it is also effective in conserving the new converts. Its small group dynamics can be contextualized in different cultures. James Engel’s “Count Down" model is effective in understanding how secular people become Christians in terms of spiritual maturity However, it overlooks the multi dimensions of how people become Christians. Agnes Riu’s “ Triangle" model is an effective way to understand the several dimensions of how secular people become Christians. It is better than James Engel’s model in this regard. However, it shows an extremely simplified conversion process. George Hunter’ s “ Target" model is an effective way to understand how secular people become Christians, also. It is an extended form of John Wesley’s “ Order of Salvation" model. It takes the c비tural gap into a serious consideration Joseph Aldrich’ s “ Life- Style Evangelism" is based on inter personal relationships. As the early church employed this model, so too modern evangelism needs to apply it to reach secular people effectively. Some evange1ism models are more effective than others according to different cultural contexts. So some models needs to be contextualized in different cultures to be effective. The Lord wants a greater harvest.

Bisphosphonates have been widely used to treat metabolic bone diseases, although the . mechanism of bisphosphonate action on bone has not been fully understood. This study aimed to examine the direct action of pamidronate on cell proliferation and differentiation of cultured human mesenchymal stem cells(hMSC). Four experimental groups and two control groups were designed; Experimental groups included both osteogenic supplement(OS) and pamidronate-treated group, pamidronate-treated group after 1 week OS treatment, only pamidronate-treated group, OS-treated group after 1week pamidronate treatrnent. Control gr。니ps included DMEMtreated group and OS-treated group. Human MSCs were isolate from bone maπow ， and cultured for 7, 14, 21 days. For the detection of osteoblastic differentiation, AI.Pase activity was measured and the expression of type 1 collagen and osteocalcin were evaluated. Von Kossa’s silver stain was performed for the examination of calcification. As results, the proliferation rate of 바1SC was maintained to be more than 90% by 1uglml of pamidronate. AI.Pase activity showed the highest value at the concentration of 100nglml of pamidronate. In pamidronate-treated group, ALPase activity reached a peak at the third week and the expression of type 1 collagen mRNA and protein was enhanced compared to other experimental and control groups, whereas osteocalcin expression was found only in OStreated group. Calcification was decreased by a dose dependent manner followed by pamidronate treatment. This study su잃，est that pamidronate treatrnent may be able to enhance the osteoblastic differentiation of hMSC at the early stage. On the other hand, calcification appeared to be inhibited by pamidronate treatrnent.

be involved in the development of oral SCC. If we compare the morphologic features of NHOK to IHOK according to calcium concentration by TEM, IHOK have been gained wide acceptance as a model system for HPV-linked oral carcinogenesis. We already have established immortalized oral keratinocytes(IHOK) transfected by E6 and E7 gene. The purpose of this study were to examined the ultrastructural features of cultured NHOK, IHOK, and HN4 oral squamous cell carcinoma cell line, and to apply these results to oral carcinogenesis in the future. NHOK from healthy retromolar pad was primarily cultured at 37oC and 5% CO2. IHOK, and HN 4 cell line which were cultured under 0.15 and 1.2mM Ca++ of KBM bullet kit. For transmission electronmicroscopy(TEM), under preconfluency, and after 3 days of postconfluency under 1.2mM Ca++, cultured NHOK, IHOK, and HN4 cell line were immediately fixed in 2.0% glutaraldehyde in 0.1M cacodylate buffer(pH 7.4) at 4OC for 1h. The ultrathin sections were stained with uranyl acetate and lead citrate, and examined by TEM. The obtained results were as follows. 1. TEM of cultured NHOK under 1.2mM Ca++ showed increased tonofilaments, and vaculated ovoid cells with cornified envelope, while cultured IHOK showed prominent microvilli, unilateral desmosome in microvillus, and tonofilaments. 2. TEM of HN 4 cell line sowed numerous microvilli, increased N/C ratio, and lateral desmosome in microvilli under 0.15mm, while under 1.2mM well forming desmosomes. From the aboving results, under high calcium cultured IHOK showed less tonofilaments than that of cultured NHOK, while cultured IHOK, and HN 4 cell lines showed more increased desmosomes under high calcium. It was suggested that the ultrastructural changes of cultured IHOK would be accepted as intermediate stage cells for studying oral carcinogenesis.