Genomic imprinting is defined as parent-of-origin expression of specific genes and may play an important role in embryonaldevelopment of mammals. Loss of imprinting(LOI), biallelic expression of the imprinted genes, have been observed in avariety of human tumors and syndromes. H19, a paternally imprinted gene, is transcribed as an untranslated RNA thatserves as a riboregulator. LOI of H19 is observed in a variety of human malignancies. In this study, LOI of H19 was examinedin head and neck squamous cell carcinomas(HNSCCs). Four(28.6%) of the 14 HNSCCs and 8(28.6%) of the 28 inflammatoryoral lesions were informative for imprinting analysis of H19. H19 was imprinted in all inflammatory oral lesions,however, 2(50%) of the 4 informative HNSCCs manifested LOI. These data suggest that LOI of the H19 may play a role in theoncogenesis of HNSCC.

Since ancient Eygypt, various dental materials were used for lost teeth including gold. The key point of this materialswere nontoxic to human body. Since early of 1990's, dental implant was done for recovery of maxillofacial defects. Frommiddle of 1970's, osseointergation concept of implant was introduced and performed in dental field. Biocompatibility of titaniumshowed good effect for osseointergration but had some problems (Galvance current and toxic corrosion) with suprastructuressuch as gold crowns. This study was performed to make safe dental implants which have reduced Galvanic currentsand corrosion. 3 kind of dental casting gold alloys (different Gold contents, 1㎝×1㎝×0.1㎝ plates.) were used as experimentalgroup, while Titanium were used as control group. Normal human osteoblasts(NHOsts)were cultured during1-4weeks for histologic study. For analysing the calcium(Ca), Phosphorus(P) and alkaline phosphatase(ALP), NHosts werecultred during 2-23days. After experiments, histologic finding were observed by LSM and SEM. Ca, P, ALP concentration byautomatic biochemical analyzer were analyzed by ANOVA test and linear regression method. The results were as follows.Biocompatibility of dental casting gold alloys were similar to titianium alloys histolgically. Biochemical analysis of dentalcasting gold alloys had no significant difference to titianium alloy except AIGIS-Fine. We could conclude that biocompatibilityof dental casting gold alloys with high contents of gold were superior to that of low contents and alloys withhigh contents of gold had no significant difference from titanium on NHost culture. Gold dental implant might be better thantitanium implant due to similar biocompatibility and no galvanic currency.

It has been reported that light-emitting diodes(LED) can be used in the treatment of oral diseases. Although bio-stimulatoryeffects of LED irradiation such as promotes stimulation of wound healing have been well known, there are few reportsabout molecular mechanism associated with cell cycle by LED irradiation. The purpose of present study was to examinethe molecular event in cell cycle of LED irradiation on primary human gingival fibroblast(hGF) in vitro. The source of lightfor irradiation was a continuous-wave LED emitting at a wavelength of 635nm, and manufactured that energy density was5mW/cm2 on sample surface. The hGF were irradiated for 1 hour at 37℃ in 5% CO2 humidified chamber. Experimental sampleswere acquired at 0 (right after irradiation), 8 and 24 hour after irradiation. To investigate the molecular mechanismsassociated with cell cycle, growth phase was determined by flow cytometry and mRNA expression of cyclin A, cyclin B, cyclinD1, cyclin E, cdc2, PCNA, p18, p27, p21, and p53 were determined by real time RT-PCR. Flow cytometric analysis demonstratedthe percentage of cells in the G1 and S phase were decreased, but the G2 phase increased, which showed cells irradiatedby LED were transitioned from S to G2 phase. For mRNA expression, cyclin B, cdc2, PCNA and p53 were increased at 0hour 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 cellsthrough transition from S to G2 phase.

Adenocarcinoma NOS of salivary glands is characterized by a high rate of local recurrences and metastasis. Long-termsurvival rate of Adenocarcinoma NOS lis not promising. Thus, different chemotherapeutical approaches had been proposedfor 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 reversethe tumor growth by targeting apoptotic pathways will be new candidates for cancer prevention and therapy. The purpose ofthis study were to study the effect of Brefeldin A(BFA) as apoptotic inducing agent in SGT cell line from human submandibularadenocarcinoma NOS and apply these results to make a plan of treatment and prognosis of salivary gland tumorsinvolving adenocarcinoma NOS. SGT cells were treated with a 300μM BFA solution in serum-free medium during 18hours. SGT cells were grown in DMEM with 10% fetal bovine serum served as controls. The growth curve and MTT assay forsuccinyl dehydrogenase activity were performed. For apoptotic analysis, fragmentation of genomic DNA was confirmed withgel electrophoresis. Transmission electron microscopy was assessed for the effect of BFA on SGT cells phenotype. Apoptoticcell 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 activityof the cells showed about 55% after 300μM BFA treatment. Destruction of cellular organells, numerous vacuolation in thecytoplasm & nucleus, chromatin margination, & fragments of nucleus were seen with TEM after 300μM BFA treatment. DNAfragmentation of SGT cell line was induced by 300μM BFA treatment and confirmed by gel electrophoresis from genomicDNA extraction. Late apoptosis of the cells through flow cytometric analysis of Annexin-V staining as induced by 300μMBFA treatment. Early apoptosis of the cells through flow cytometric analysis of caspase 3 and Apo 2.7 staining was inducedby 300μM BFA treatment. It suggested that early and late apoptosis of SGT cell line would be induced by Brefeldin A treatmentin vitro study. This work evaluated the efficacy of BFA, a potent apoptosis inducer, on SGT cultured cell line. And BFAas chemotherapeutic agent will be used as the treatment choice for adenocarcinoam NOS, and be need to apply BFA to invivo study & clinical approach in future.

Several tumor animal models have been provided as a tool for developing cancer therapy. Here, we developed rapid,easy-to use, and cost-effective new rat animal model for invasion and metastasis of cancer using genetically k-ras-inducedrat kidney cells(RK3E-ras). We observed tumor as early as 3 days after injection of RK3E-ras cells in subcutaneous ofSprague-Dawley rats. Tumor size and volume were increased exponentially for 2 weeks. The tail vein injected rats obtainedthe lethal infiltration in the lung within 2 weeks. This tumor animal model has great potential for studying cancer processesand short-term screening of variable cancer therapy strategy.

A novel indirubin analog, 5'-nitro-indirubinoxime inhibits cell proliferation and induces apoptosis against various humancancer cells. In this study, we performed the microarray analysis to identify genes differentially expressed in the KB oralsquamous carcinoma cells after treated with 5'-nitro-indirubinoxime. Among the 10,800 genes analyzed, 1,701 genes (15.8%)showed statistically different expression level in the 5'-nitro-indirubinoxime treated cells with respect to untreated controlcells. Among those, 263 genes (15.5%) were down-regulated and 220 genes (12.9%) were up-regulated more than 2-fold.Functionally related gene clusters include genes associated with signal transduction (18.1%), especially genes related withapoptosis (3.5%) and cell cycle regulation (5.8%). Our application of microarray analysis on 5'-nitro-indirubinoxime treatedoral cancer cells allows the identification of candidate genes for providing novel insights into the indirubin mediated antitumoractivity.

The purpose of this study was to evaluate the role of Fas, Fas-L, and FAP-1 expression in the oral squamous cell carcinomasand ameloblastomas. For this study, 10 subjects diagnosed as squamous cell carcinoma and 8 subjects of ameloblastomareferred to the Dept. of Oral Pathology, School of Dentistry, Kyung Hee University, 5 subjects of normal oral mucosawithout any inflammatory changes were used as experimental, control groups respectively. All the tissues ; experimentaland control group were fixed in neutral formalin solution and embedded in paraffin, serial tissue section weremade 5㎛ in thickness and processed in the standard way for immunohistochemical method, using primary antibody againstFas, Fas-L, FAP-1, each was diluted at 1;100 followed by the super sensitive non- biotin horse radish peroxidase detectionsystem with DAB as chormogen, counterstained with Gill's hematoxylin stain method , mounted. And examined under thebiologic microscope with the criteria of -(no epithelial stain), +(weak or focal epithelial stain), ++(moderate or focal intensiveepithelial stain), +++(intense generalized epithelial staining) for the epithelial, and connective tissue component insquamous cell carcinomas , ameloblastomas and normal oral mucosa on each. In normal oral epithelium, negative reactionwas noted on the Fas . Fas-L stain, but on FAP-1 reaction, tumors cells with intense reaction on nuclei and cytoplasm ornegative reaction on nuclei with intense reaction on cytoplasm were admixed. On Fas, Fas-1 reaction, both tumor cells ofameloblastoma and oral squamous cell carcinoma showed negative reaction on nuclei and cytoplasms. On FAP-1 reaction,tumor cells of oral squamous cell carcinomas showed more intensive response compare to that on ameloblastomas.Considering these results, the tumor cells of ameloblastoma and squamous cell carcinoma showed negative reaction on theFas and Fas-L, but it could suggest that FAP-1 induce the development of tumors by means of inhibition of the apoptosis.

Cyclooxygenase(COX)-2 is an enzyme engaged in the synthesis of prostaglandin(PG) and is upregulated by inflammation.It is related with the expression of the vascular endothelial growth factor(VEGF) in the angiogenesis of granulation tissue.By using immunohistochemical stains, the expressions for COX-2 and VEGF were evaluated according to the degree of inflammationand the type of lesion, and the correlation between the expressions were investigated in the 39 periapical lesions(7 periapical granulomas, 7 periapical granulomas with epithelium, 25 periapical cysts) and 13 dentigerous cysts. The expressionrate of COX-2 in histiocytes of periapical lesions(97.4%) was significantly higher than that of dentigerouscysts(69.2%)(p=0.0032). The expression rate of VEGF was highest in plasma cells in the periapical lesions(69.4%) and indentigerous cysts(69.2%). COX-2 expression rate in endothelium and VEGF expression rate in epithelium were significantlyhigher in the periapical cysts than in the periapical granuloma and. VEGF expression rate in endothelium of the periapicallesions was increased in cases with resolving inflammation. For the periapical lesions, epithelial expression of COX-2 andVEGF showed a positive relationship(p=0.0301), and endothelial COX-2 expression revealed positive relationship with VEGFexpressions in epithelium(p=0.0008), in histiocytes(p=0.0136), and in endothelium(p=0.0439). According to these findings, asthe periapical lesion progressed from granuloma to cyst, the expressions of COX-2 and VEGF were increased. COX-2 expressionwas significantly correlated with VEGF expression, which suggests that COX-2 seems to be involved in the progressionof periapical lesion by inducing the expression of VEGF.

Angiosarcoma is an extremely rare sarcoma arising from oral mucosa. We report a case of angiosarcoma in the rightmaxillary gingiva causing excessive bleeding. The lesion exhibited typical histologic features of angiosarcoma, showinginfiltrative proliferation of polygonal endothelial cells and arborizing blood vessels. Tumor cells showed expression ofCD 31 and factor VIII, but no expression of CD 34 antigen. The patient expired due to severe bleeding in the oralcavity.

In order to investigate stresses of dental patients in dental clinic by dental-analogous stimulations, we selected 23women and 36 men who are students of dental college in D university. This experiment was performed to compare andanalyze the changes of Galvanic skin resistance and heart rate by dental-analogous stimulations. In paired t-test ofmale group's GSR average, there were significant differences between sound and touch, and among pain and otherstimulations. And in paired t-test of female group's GSR average, there were significant differences among pain andother stimulations. In paired t-test of male group's heart rate average, there were significant differences between visionand pain, and between touch and pain. And in paired t-test of female group's heart rate average, there were significantdifferences between smell and pain. In unpaired t-test of male and female group's GSR average, there were nosignificant differences in smell, sound, vision, touch, and pain. In unpaired t-test of male and female group's heartrate average, there were no significant differences in smell, sound, vision, touch, and pain. It seemed that stressesduring dental treatment could be changed by surrounding circumstances and sex distinction.

Immuno-MemBlot is a technique for detecting, analyzing, and identifying proteins, similar to the Western blot techniquebut differing in that protein samples are not separated electrophoretically but are spotted through circular orslot templates directly onto the membrane. Recently we developed a new Immuno-MemBlot (IMB) method applying immunoreactionsand coloring procedures directly in the wells of MemBlot apparatus, which were connected by canals toperform drainage for reagent application and buffer irrigation. This IMB method was designed to get theimmunoblotresults more rapidly and clearly than the previous immunoblot ones. This study is aimed to evaluate the analytical accuracyof IMB using different biological assay. In the sensitivity test of IMB the monoclonal antibody can clearly detectthe 30 ng (about 12 pM) of Mucocidin peptide (35 mer), and is also available to detect at least 10 ng (about 4 pM) ofMucocidin peptide (35 mer). The IMB was effective in the quantitative analysis of methothrexate (MTX) assay for cellularapoptosis. And more, this IMB is useful to screen large number of specific samples with ease and accuracy in ashort time. In the screenings for the presence of Mucocidin in saliva the quantitative comparison is conspicuous among48 persons depend on the different conditions ofgender, drinking and smoking habits, and oral diseases. Therefore, itis presumed that, even though the target proteins were partly degraded, a specific epitope can be detected if a monoclonalantibody was still reactive. Conclusively, these data suggest that the IMB can be useful in the primary qualitativeandquantitative analysis of proteins in various fluids, i.e., blood, saliva, tear, urine, etc.

The periodontal ligament (PDL) is that soft, specialized connective tissue situated between the cementum coveringthe root of the tooth and bone forming the socket wall. The PDL is a connective tissue particularly well adapted to itsprincipal function, supporting the teeth in their sockets and at the same time permitting them to withstand the considerableforce of mastication. During the life time, PDL is usually exposed to mechanical stress by mastication.However, little is known about the gene which is related to the mechanical stress in PDL. UNC-50 (PDLs22) was identifiedand isolated from D. melanogater and C. elegance. This gene was also regulated in sensory bristle for mechanotransductionin D. melanogaster. In this study, to uncover the relationship between UNC-50 and mechanical stress, we induced the mechanicalstress by medium displacement in cementoblast cell line. After mechanical stress induction UNC-50 expression wasanalyzed by RT-PCR, Real-time PCR, and western analysis. The expression of UNC-50 was increased after medium displacementof cementoblast in vitro. Collagen type I, type III, and osteonection mRNAs were also strongly expressed aftermechanical stress induction. The results of this study suggest that UNC-50 might responsible for molecular event in PDLinducing cementoblast under mechanical stress.