Cytokines play a vital role in the host immune response by regulating the development and function of im munocompetent ce11s One immunomodulatory agent that has received attention in oncology research recently is interleukin - lO(IL-lO). IL-IO inhibi ted tumor antigen presenta tion and induced energy in T lymphocytes that had been s timu lated by autologous MHC class II positive tumor ce11s Patients with head and neck cancer have been shown to exhibit profound irnmunosuppression. The mechani sm by which tumor ce11s alter immunological function in the host is poorly understood. Recently. production of biological active IL- IO was confirmed in ovar‘ian cancer, melanoma, skin cancel‘ & head and neck cancer, suggesting that IL- lO reduces the function of tumor infiltrating lymphocytes and contributes to the tumor growth. IL-IO expression has not been examined extensively in human oral cancer and has not yet been cla rified. The purpose of t his study were to investigate IL-IO mRNA and protein expression in NHOK, IHOK and oral squamous ce11 carcinoma(OSCC) ce11 line by RT-PCR and irnmunoslot blotting, and to apply its results to examine its thera peutic significance for oraJ cancers. Cultured NHOK showed a lower level of IL-IO mRNA and protein expression than cultured IHOK and HN 22 OSCC cell line under pre and postconfluency. HN 22 OSCC cell line under pre and postconfl u ency. showed the highest level of IL-I0 Cul tured IHOK showing a intermediate expression of IL- IO could be as a vaJ u a bJe marker for oral carci nogenesis ste p. During the terminal differentiation of a11 the ce11 lines, IL- IO ex pression was significantly unchangeabl e. IL- IO mRNA expression of a11 the ce11 lines was consistent with IL-10 protein expression. It suggested that IL- lO expression might play an important role in oral carcinogenesis and IHOK could be a valuable marker for oral carcinogenesis step. And aJso IL- 10 related gene may be future targets for gene discovery and possi bJy therapeutic intervention

Desmoplastic ameloblastoma(DA) is histologically characterized by extensive stromal collagenization or desmoplasia. ln this study, anti-cytokeratin 8/18, 13, 19 for pathogenesis as well as anti-PCNA for cellular proliferation, were used to det ect the expression of these proteins in the desmoplastic ameloblastoma Basal layers of tumor nest were negatively stained by CKl3, while suprabasal and inner cells were positive for CK13. CK8/18 and CK 19 was negatively stained in the peripheral portion of tumor nest in DA, whereas CK 8/18 was in central portion and CKl9 was positive in the su prabasal and some of central portion of the cel l nest. PCNA index of DA was 60 ::!: 14.6% to 95 ::!: 17 .2%. The peripheral tumor cells of the islets presented higher PCNA labeling index, while some cells in the central area of foll icle containing squamous like cells also presented negative PCNA labeling index. Especially tumor islands showed higher PCNA index than in main tumor mass. lt suggested that desmoplastic ameloblastoma might be composed of many different tumor cell types‘ and have hi gher pr이 ife r a ting activity in tumor islands of the desmoplastic stroma

This r esearch was designed to find the specific and economic methods of diagnosis about malignant melanoma For this study, we selected a typical case that was ambiguous in diagnosis between malignant melanoma and simple mela notic pigmentation, Tissue sections was st ained by H&E method, and immunohistochemical analyses was performed about 8-100 protein and MART-1 molecule, This research showed that MART-l had a more specificity for melanocytes than 8-100 protein , Patterns of MART-1 molecule distribution was more helpful in estimation of malignancies than 8-100 protein distribution patterns, On the basis of diagnostic usefulness and economical aspects, 8-100 protein and MART-1 molecule showed synergis tic and complementary relationship in confirming of tumor origin and they would be much useful for accurate diagnosis of malignant melanoma

Extensive oral mucosa loss from a variety of conditions is associated with significant functional morbidity and mortality. Although it is known that keratinocytes are a rich source of wound healing promoting factors such as transforming growth factor-β1(TGF-β1), it is not clear whether differentiated keratinocytes in a multi-layer form release this multi-functional growth factor. This study examined the hypothesis that keratinocytes in mono- and multi-layer forms expressed different levels of TGF-β1. When NHOK reached confluency in serum free medium(KBM), in test medium containing 1.2 mM Ca++ KBM NHOK were allowed to form multi-layers and differentiate. The purpose of this study were to investigate the mRNA level of TGF-β1, FGF-2, and TIMP-1 by RT-PCR analysis and also to evaluate the expression of TGF-β1 and involucrin in keratinocytes at different times of the onset of differentiation. The numbers and sizes of these nodules were increased as the process of keratinocyte differentiation proceed. Cultured NHOK in preconfluency under KBM medium expressed a significantly higher level of TGF-β1 relative to those grown in multi-layer forms, while the level of TGF-β1 mRNA gradually reduced to its lowest level at 7 days of growing cells in test medium. Cultured NHOK in preconfluency of KBM medium expressed a lower level of FGF-2 and TIMP-1 relative to those grown in multi-layer forms, while the level of FGF-2 and TIMP-1 mRNA showed the highest level at 3 days at gradually reduced to its lowest level at 7 days of growing cells in test medium. As a differentiation marker for keratinocytes at different time points, the highest level of involucrin mRNA expression was found at the later stage of cell differentiation. It suggested that the expression of TGF-β1 mRNA be consistent with the expression of FGF-2 and TIMP-1 mRNA in NHOK grown in high calcium medium during the terminal differentiation. But differentiated NHOK expressing higher involucrin mRNA could show constant espression of TGF-β1, FGF-2 and TIMP-1.

Annexin I plays an important role in the process of keratinization as a compont of the cornified envelope of skin epithelium. The effect of annexin I on the terminal ifferentiation of normal human oral keratinocyte(NHOK) have remained to be defined. To understand the role of annexin I on the terminal differentiaiton of NHOK, NHOK and NHEK cells were primarily cultured in KBM bullet kit. When the cells reached confluence, terminal differentiation was induced by switching the medium to KGM bullet kit containing 1.2mM Ca2+. Preconfluency of NHOK under 0.05mM Ca++ conc as control group was used. The cells was examined with inverted microscope. Under 0.05mM Ca++ conc(Precon, Postcon), and 1.2mM Ca++ conc(Postcon), RT-PCR for annexin I mRNA measurement, and immunoblotting for annexin I protein measurements in triplicate, respectively. The purpose of this study were to study differential mRNA & protein expression of annexin I between NHOK & NHEK by using RT-PCR & immunoslot blotting during terminal differentiation, and to apply these results to study a role of annexin I on epithelial differentiation of oral mucosal diseases in the future. Cultured NHEK showed larger area of cellular stratification than cultured NHOK in 1.2mM Ca ++ concentration. Annexin I mRNA and protein expression of cultured NHOK showed higher than that of cultured NHEK in higher calcium concentration. Annexin I mRNA and protein expression of cultured NHOK showed about 2-2.7 fold higher in 1.2mM Ca++ conc. than in 0.05mM Ca++ conc. Although annexin I was involved in the terminal differentiation of cultured NHOK & NHEK in higher calcium concentration, annexin I play an important role in the terminal differentiation of cultured NHOK in higher calcium concentration. From the aboving results, It was suggested that annexin I would play an important role in the terminal differentiation of NHOK in higher calcium, which be helpful to study epithelial differentiation of oral mucosal diseases.

Pro-inflammatory cytokines are important mediators of cutaneous cellular activities during many oral mucosal diseases. IHOK culture model transfected by E6/E7 genes provide further evidence for the role of HPV in tumorogenesis. It is interesting to investigate cytokine expression of immortalized human oral keratinocyte(IHOK). The purpose of this study were to analysis cytokine mRNA expression levels of NHOK and IHOK by RT-PCR. IHOK showed about 5 fold increases of IL-6 compared with NHOK, while TNF-α was the lowest. It suggested that immortalization of NHOK with E6/E7 could result in elevated expression of IL-6, and IHOK be in the intermediate stage of oral carcinogenesis.

Adenocarcinoma NOS of salivary glands is characterized by a high rate of local recurrences and metastasis. Long-term survival rate of Adenocarcinoma NOS lis not promising. Thus, different chemotherapeutical approaches had been proposed for this neoplasm, including apoptosis induction by drugs. The current treatment of choice of adenocarcinoma NOS is controversible, and an effective treatment for them is not yet available. Chemotherpeutic agents that can be inhibit or reverse the tumor growth by targeting apoptotic pathways will be new candidates for cancer prevention and therapy. The purpose of this study were to study the effect of Brefeldin A(BFA) as apoptotic inducing agent in SGT cell line from human submandibular adenocarcinoma NOS and apply these results to make a plan of treatment and prognosis of salivary gland tumors involving adenocarcinoma NOS. SGT cells were treated with a 300μM BFA solution in serum-free medium during 18 hours. SGT cells were grown in DMEM with 10% fetal bovine serum served as controls. The growth curve and MTT assay for succinyl dehydrogenase activity were performed. For apoptotic analysis, fragmentation of genomic DNA was confirmed with gel electrophoresis. Transmission electron microscopy was assessed for the effect of BFA on SGT cells phenotype. Apoptotic cell recognition and counting were carried out with Annexin-V, caspase 3 and APo2.7 antibody through flow cytometry. Growth of SGT cell line was abrutply decreased after 1 day of BFA treatment. MTT assay for succinyl dehydrogenase activity of the cells showed about 55% after 300μM BFA treatment. Destruction of cellular organells, numerous vacuolation in the cytoplasm & nucleus, chromatin margination, & fragments of nucleus were seen with TEM after 300μM BFA treatment. DNA fragmentation of SGT cell line was induced by 300μM BFA treatment and confirmed by gel electrophoresis from genomic DNA extraction. Late apoptosis of the cells through flow cytometric analysis of Annexin-V staining as induced by 300μM BFA treatment. Early apoptosis of the cells through flow cytometric analysis of caspase 3 and Apo 2.7 staining was induced by 300μM BFA treatment. It suggested that early and late apoptosis of SGT cell line would be induced by Brefeldin A treatment in vitro study. This work evaluated the efficacy of BFA, a potent apoptosis inducer, on SGT cultured cell line. And BFA as chemotherapeutic agent will be used as the treatment choice for adenocarcinoam NOS, and be need to apply BFA to in vivo study & clinical approach in future.

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.

Odontomas are the most common benign tumors of odontogenic origin characterized by their‘ slow growth, and a re cons ider ed to be developmental a nomalIes They consist of enamel, dentine, cementum and pulpal tissue and cons titue about 22% of all odontogeni c tumors The purpose of this study was to value clinicopathologic features of this kind 01 tumors‘ and to apply these results to make a pa ln of diagnosis and treatment in future, The 20 cases of odontomas between 1991 a nd 2004 subrn.i tted to Dept , of Oral Pathology of Dankook Univ, Dental Hos pital were used We analyzed theil‘ prev a lence‘ di s tributi on in gender , age of the pat ients , anatornic location. clinical manifestations and histopa thologic fea rtures Cli nical symptoms s howed delayed eruption(n=3) of either the deciduous or permanent tooth‘ mtra- 01 extraoral swelli ng(n =9). pain(n =2) , Of a ll the odontomas, 45% in the Mn & 55% in the Mx was seen There were 55% in the anter iosuperier sector & 25% i n the pos ter ioi nferior zone. while the rest in the upper molar region, COO showed 18,5 year s(r ange 6,4) , 60% 0[' diagnosed odontomas, incisor / canine area of mx, smaller(5-30mm) & unilocular. and mu l tip le radiopaque mini teeth s urroun따d by defined radiolucency, whi le CXO showed 30, 1 years(range 14 , 5) ‘ 40% of diagnosed odon tomas ‘ 1s t and 2nd molar of the mn, la rger(1O-60mrn) & unilocular undistinguished radiopaque mass and surrounded by ra di 이 uc e nt zo ne & a di s tin ct ra di opaque border, Both showed reduced enamel epithelium, enamel. dentin and cementum‘ di s play ing s ph eri al calcificat ions. and fibrous capsule as background , A.nd ghost cells were seen in 2 cases COO s howed hig hly orga nization & differenti ation, resembling a minitooth Each tooth was contained with co n nective tissuc sac CXO s howed haphazard organization, and was domi nated by dentin , displaying no resemblance a tooth, Through the definitive diagnos is of odonotmas on the basis of the clinicopath이 ogi c results, we should establish a plan in diagnosis and t rea tment of odontomas

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

Dental problems cause not only esthetic but dental problems such as functional disorder, dental caries, pulp disease and malocclusions, and in particular, masticatory problem falls on infants or children, which has effect on articulation disorder and emotional development as well as their physical growth. Therefore, it's important to detect dental problems early and to provide comparable indications. The purose of this study was to find out the prevalence and pattern of dental developmental anomalies. The clinical and radiographic examination was undertaken for 1,032 at age from 1 to 10 years and statistical analysis was done. The result were as follows. Among the examined patients, 333 patients( 32.3%) showed dental developmental anomalies. The prevalences of individual dental anomalies were as follows ; congenital missing teeth 11.6%, supernumerary teeth 6.2%, taurodontism teeth 3.9%, fusion teeth 3.8%, microdontia teeth 3.2%, dens evaginatus teeth 2.3%, dens invaginatus teeth 0.6%, talon cusp teeth 0.2%, macrodontia teeth 0.1%, Dilaceration teeth 0.1%, transposition teeth 0.1%, inverted teeth 0.1%, amelogenesis imperfecta 0.1%.. The anomalies of number(53.6%), the anomalies of shape(37.7%,) the anomalies of size(8.3%), the anomalies of position(0.3%) and the anomalies of structure(0.2%.) were examined. Most supernumerary teeth located on middle. area showed inverted position and unerupted state. The most frequently missing teeth was the mandibular primary lateral incisor(65.6%) and the mandibular 2nd. premolar. In the case of deciduous congenital missing teeth, the prevalence of successive permanent congenital missing teeth (65.6%) was highest in mandibular lateral incisors.. The higher prevalence of microdontia teeth was maxilla lateral incisor, dens evaginatus teeth was mandibular 2nd. premolar, dens invaginatus teeth was maxilla lateral incisor, and taurodontism was mandibular 1st. molar. Teeth play an important role for mastication, articulation, and aesthetic of adults as well as of children. The health of oral cavity could be kept by controling dental problems of children in the basis of the epidermiologic research of oral pathologic field.

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.

It is very little known that the molecular mechanisms control growth, cell differentiation, and invasion of ameloblastoma into bone. Tissue culture methods have also been used extensively for studies of the cell biology of ameloblastoma. The purpose of this study were to examined the ultrastructural features of ameloblastoma, and to apply these results to examine the pathogenesis of ameloblastoma in the future. Amelobalstoma was primarily cultured under 0.1, 0.15 and 1.2mM Ca++ of KBM bullet kit at 370C and 5% C02. For transmission electronmicroscopy(TEM), cultured ameloblastoma cells were immediately fixed in 2.0% glutaraldehyde in O.lM cacodylate buffer(pH 7.4) at 40C for 1h The ultrathin sections were stained with uranyl acetate and lead citrate, and examined by TEM. The obtained results were as follows. Primay culture ameloblastoma grown in 0.1 mM Ca++ showed interlacing papillary projections without desmosome within early passage(3-4). Primary culture amelobalstoma under high calcium showed prominent desmosomes or tight junctions within early passage. There was evidence of cellular degeneration, as exemplified by nuclear pyknosis, the margination and clumping of the chromatin, and vaculolation under high calcium. The sparse ribosomes, the cytoplasmic space filled with vacuoles, and the condensed mitochondria were seen under high calcium. From the aboving results, under high calcium primary culture ameloblastoma showed rapid cellular degeneration within early passage, indicating that the cells were gradually losing metabolíc actívitíes, leading to enventual cell death. It was thought that it would be necessary to establish cultured immortalized amelobalstoma cell line for studying the pathogenesis of odontogenic tumors.

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,

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.

Transglutaminase 2(TGase 2) expression is modulatecl by π>JF- a in various carcinoma. The role of TGase 2 ancl TNF- a expression in salivaIY gland tumors is not clear yet. Establishecl SGT cellline has been used to study the pathogenesis of salivaIY gland adenocaI‘cinoma on a cellular level in vitro. 까le pupose of this study were to examine n버NA expression of TGase 2 and TNF- ain SGT cellline comparecl to other tumor celllines, ancl to apply these results to the paùlogenesis of salivary gland tumor. After SGT, SCC-15, HN 4, and HeLa tumor celllines were culturecl under preconfluency, ancl 3 clays after postconfluency, the cells were harvested for total RNA extraction and cDNA preparation. RT-PCR for semiquantitative mRNA analysis was done. 까le obtained results were as follows. 1. TGase 2 and π>JF- amRNA expression was not induced by confluency in all the celllines 2. TGase 2 and π'JF- amRNA expression was variable but markeclly enhanced 비 SGTcellline 3. TGase 2 n버NA expression appeared to be associated with that of π>JF- ain SGT cellline From the aboving res ults, mRNA expression of TGase 2 and TNF a should play an important role in the pathogenesis of SGT cellline originated ti'om ductal cell.

It is well known that glycoproteins, glycosaminoglycans and proteoglycans are of fllndamental importance to the processes of morphogenesis and cytodifferentiation dllring the teeth development. With HID-TCH-SP(High-iron diamine-thiocarbohydrazide-silver proteinate), slllfated glycosaminoglycans sllch as chondroitin slllfate and heparan slllfate have been localized at the 1I1trastructurallevel in a wide variety of tisslles. 까le pllrpose of this study were to characterize slllfated glycosaminoglycan profiles of hllman fetal tooth genns at 비trastructurallevel for the phase of morphogenesis and cytodifferentiation, and to detect the protein expression of sulfated glycosaminoglycan by immunoslot blot. Human tooth germs from the alveolar bone of twenty still born fetuses were f1xed in a mixture of 2% glutaraldehyde/ l% fonnaldehyde. The ultrathin sections were stained with HIDTCH- SP, and some sections were σ'eated with 0.05% solution of testicular hyaluronidase to identify the histochemical properties of the HID-TCH-SP stain deposits. For semiquantitative protein assay, immunoslot blot was done. The obtained results were as follows 1. HID-TCH-SP staining showed sulfated glycocongugated deposits in DEJ, peritublllar dentin, and mantle dentin matrix, enamel prism sheath, interrod area, and enamel matrix. 2. Heparan sulfate deposits in DEJ resisted to testicular hyalllronidase treatment prior to HID-TCH-SP staining 3. In immunoslot blot, chondroitin slllfate was detected higher in enamel and dentin extraα ， while heparan slllfate was relatively expressed in enamel and dentin extract, but rarely expressed in enamel or dentin extract. From the aboving results, it was suggested that chondroitin and heparan sulfate would play an important role in the formation of D티， while chondroitin sulfate would in the development of enamel prism sheath, enamel matrix, and mantle or peritllblllar dentin of human fetal t∞th germs.