Though hydrogen peroxide (H₂O₂) causes a deleterious effect to cells with its reactive oxygen species resulting in cell death, S-allyl cysteine (SAC, a bioactive organosulfur compound of aged garlic extract) has been known to have a cytoprotective effect. Few reported profiles of gene expression of H₂O₂ and SAC treated human cord blood derived mesenchymal stem cells (MSC). This study revealed changes in the profile of twenty-one genes grouped by oxidative stress, antioxidant, cell death, anti-apoptosis and anti-aging by quantitative real time PCR. A concentration of 100μM of SAC or 50μM of H₂O₂ was applied to MSC which show moderate growth and apoptosis pattern. H₂O₂ treatment enhanced expression of eleven genes out of twenty-one genes compared with that of control group, on the contrary SAC suppressed expression of eighteen genes out of twenty-one genes except C ros oncogene. SAC decreased expression of oxidative stress genes such as SOD1, CAT and GPX. These results seemed consistent with reports which elucidated over expression of NF-kB by H2O2, and suppression of it by SAC. This study will confer basic information for further experiments regarding the effects of SAC on gene levels.

Resveratrol (RVT) and epigallocatechin gallate (EGCG) individually inhibit adipogenesis in 3T3-L1 adipocytes. The objective was to examine the possibility of interaction between RVT and EGCG, resulting in enhanced inhibition of adipogenesis in 3T3-L1 adipocytes. Preadipocytes were treated with RVT and EGCG individually at 6.25 or 25μM (RVT6.25 or RVT25) and 12.5 or 50μM (EGCG12.5 or EGCG50) and in combination (RVT6.25 + EGCG12.5 and RVT25 + EGCG50). RVT25 as an individual compound decreased lipid accumulation in 3T3-L1 adipocytes by 24%, and RVT25 + EGCG50 further decreased lipid accumulation by 77%. In addition, exposure of 3T3-L1 adipocytes to RVT6.25 + EGCG12.5 and RVT25 + EGCG50 combinations resulted in an enhanced increase of adiponectin release and inhibition of leptin release. Quantitative analysis revealed that the combination of tested materials (RVT6.25 + EGCG12.5 and RVT25 + EGCG50) decreased the expression levels of C/EBPα, PPARγ2, and aP2. These results indicate that the combined treatments with RVT and EGCG produce synergistic effects on inhibiting adipogenesis in 3T3-L1 adipocytes. The overall results suggested that the combining RVT and EGCG might be more capable of exerting antiobesity effects than each individual compound by itself.

영지버섯 자실체를 열수추출과 주정추출을 하여 항산화 효능 및 간암 및 위암세포의 생장 저해도를 분석하였다. 항산화 효능을 실시한 결과, 대조군인 Trolox, BHA보다 높은 항산화 효능을 보인 것은 ASI 7004, 7014이며, 이 두 균주는 열수추출물과 주정 추출물에서 모두 높은 항산화 효능을 나타냈다. 나머지 균주는 대조군인 ABTs보다 대체적으로 높은 효능을 보였다. 또한 암세포 생장저해도를 알아보기 위해서 열수, 주정 추출물을 간암세포인 HepG2에 농도별로(100, 200, 400μg/ml) 처리하여, 세포 생장 저해도(MTT assay)를 측정한 결과, 열수 추출물에서는 ASI 7002, 7011, 7014, 7020이 농도 의존적으로 간암세포의 생장을 저해하는 것을 알 수 있었다. 또한 주정 추출물에서는 ASI 7011, 7019가 농도 의존적으로 간암세포 생장을 저해하는 것으로 나타났다. 위암세포인 AGS에 농도별로(100, 200, 400μg/ml)처리하여, 세포 생장 저해도(MTT assay)를 측정한 결과, 열수 추출물에서는 ASI 7001, 7002, 7019, 7020이 농도 의존적으로 위암세포의 생장을 저해하는 것을 알 수 있었다. 또한 주정 추출물에서는 ASI 7001, 7002가 농도 의존적으로 위암세포 생장을 저해하는 것으로 나타났다.

Osteoarthritis is one of the commonest causes associated with age-related damage of articular cartilage. Non-steroidal anti-inflammatory drugs are commonly used in osteoarthritic patient. However, long term administration of these drugs results gastrointestinal disorders. Though, most studies have demonstrated in the past that bee venom has therapeutic effect on diseases related to inflammation and pains, but its anti-inflammatory properties have not been so far studied on inflamed chondrocytes (LPS induced) invitro. For the purpose, the study was carried out to determine the effect of bee venom on porcine articular chondrocyte cell using microarray. In this study, we found that 2,235 significantly associated gene (1,404 up-regulated genes and 831 down-regulated genes) that were expressed on inflamed and non inflamed chondrocytes during proliferation. Among the 1,404 up-regulated genes and 831 down-regulated genes, known genes were 372 and 237, respectively. On the other hand, bee venom significantly reduced expression of fetuin involved in acute inflammatory reaction. Our results suggest that this study could be useful database in gene expression profiling of chondrocyte cell treated with bee venom.

Abstract. The keratoameloblastoma is a benign lesion of the jaws, which is a rare histologic variant of the ameloblastoma. There is a variation in the histopathologic appearance of reported cases under the appellation of keratoameloblastoma. The keratoameloblastma has usually keratin formation by the ameloblastomatous epithelium and varies in size. English literature reports only 14 cases of keratoameloblastma. We described an additional case of the tumor developed in the right mandible of a 26-year-old woman. It was presented as an expansile and radiolucent lesion. Histologically, solid tumor islands were seen with some microcystic space within a fibrous stroma resembling an ameloblastoma. In addidion, a hyperchromatic columnar basal cell layer and parakeratin within the microcyst simulating an odontogenic keratocyst

Although the mechanism of the abnormal calcification in the calcifying odontogenic cyst (COC) was not elucidated so far, it has been known that the ghost cells are closely related to the calcification, producing dystrophic globular cementum-like materials, comparable to pilomatricoma in epithelium1). Here, we presented a case of COC occurred in left maxillary canine area of 23 years old female, exhibiting a collection of aberrant ossification admixed with basophilic ghost cells in comparison with seven cases of COC. In the polarizing microscope observation with Masson trichrome stain the present case clearly disclosed the typical birefringence of bony tissue, stained red in von Gieson stain, indicating the collagenous backbone. Some ghost cells showed the features of interdigitating epithelial attachments, empty spaces of nuclei, and reticular basophilic cytoplasms, which were similar to the basophilic ghost cells of philomatricoma. The present case demonstrated the aberrant ossification by basophilic ghost cells in COC similar to the ossification of pilomatricoma

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.

Adult stem cell transplantation has been increased every year, because of the lack of organ donors for regenerative medicine. Therefore, development of reliable and safety cryopreservation and bio-baking method for stem cell therapy is urgently needed. The present study investigated safety of dimethyl sulfoxide (DMSO) such as common cryoprotectant on porcine bone marrow derived mesenchymal stem cells (pBM-MSCs) by evaluating the activation of Caspase-3 and -7, apoptosis related important signal pathway. pBM-MSCs used for the present study were isolated density gradient method by Ficoll-Paque Plus and cultured in A-DMEM supplemented 10% FBS at in 5% incubator. pBM-MSCs were cryopreserved in A-DMEM supplemented either with 5%, 10% or 20% DMSO by cooling rate at /min in a Kryo 360 (planner 300, Middlesex, UK) and kept into . Survival rate of cells after thawing did not differ between 5% and 10% DMSO but was lowest in 20% DMSO by 0.4% trypan blue exclusion. Activation of Caspase-3 and -7 by Vybrant FAM Caspase-3 and -7 Assay Assay Kit (Molecular probes, Inc.OR, USA) was analyzed with a flow cytometer. Both of cryopreserved and control groups (fresh pBM-MSCs) were observed after the activation of Caspase-3 and -7. The activation did not differ between 5% and 10% DMSO, but was observed highest in 20% DMSO. Therefore 5% DMSO can be possibly used for cell cryopreservation instead of 10% DMSO.

The synovial tissues are a valuable MSCs source for cartilage tissue engineering because these cells are easily obtainable by the intra-articular biopsy during diagnosis. In this study, we isolated and characterized the canine MSCs derived from synovial fluid of female and male donors. Synovial fluid was flushed with saline solution from pre and post-puberty male (cM1-sMSC and cM2-sMSC) and female (cF1-sMSC and cF2-sMSC) dogs, and cells were isolated and cultured in advanced-DMEM (A-DMEM) supplemented with 10% FBS in a humidified 5% atmosphere at . The cells were evaluated for the expression of the early transcriptional factors, such as Oct3/4, Nanog and Sox2 by RT-PCR. The cells were induced under conditions conductive for adipogenic, osteogenic, and chondrogenic lineages, then evaluated by specific staining (Oil red O, von Kossa, and Alcian Blue staining, respectively) and analyzed for lineage specific markers by RT-PCR. All cell types were positive for alkaline phosphatase (AP) activity and early transcriptional factors (Oct3/4 and Sox2) were also positively detected. However, Nanog were not positively detected in all cells. Further, these MSCs were observed to differentiate into mesenchymal lineages, such as adipocytes (Oil red O staining), osteocytes (von Kossa staining), and chondrocytes (Alcian Blue staining) by cell specific staining. Lineage-specific genes (osteocyte; osteonectin and Runx2, adipocytes; PRAR-, FABP and LEP, and chondrocytes; collagen type-2 and Sox9) were also detected in all cells. In this study, we successfully established synovial fluid derived mesenchymal stem cells from female and male dogs, and determined their basic biological properties and differentiation ability. These results suggested that synovial fluid is a valuable stem cell source for cartilage regeneration therapy, and it is easily accessible from osteoarthritic knee.

Various small molecules can be used to control major signaling pathways to enhance stemness and inhibit differentiation in murine embryonic stem cell (mESC) culture. Small molecules inhibiting the fibroblast growth factor (FGF)/ERK pathway can preserve pluripotent cells from stimulation of differentiation. In this study, we aimed to evaluate the effect of pluripotin (SC-1), an inhibitor of the FGF/ERK pathway, on the colony formation of outgrowing presumptive mESCs. After plating the zona pellucida-free blastocyst on the feeder layer, attached cell clumps was cultured with SC-1 until the endpoint of the experiment at passage 10. In this experiment, when the number of colonies was counted at passage 3, SC-1-treated group showed 3.4 fold more mESC colonies when compared with control group. However, after passage 4, there was no stimulating effect of SC-1 on the colony formation. In conclusion, SC-1 treatment can be used to promote mESC generation by increasing the number of early mESC colonies.

The key regulators of apoptosis are the interacting protein of the Bcl-2 family. Bcl-2, an important member of this family, blocks cytochrome C release by sequestering pro-apoptotic BH3-only proteins such as Bid, Bad, Bax and Bim. The pro-survival family members (Bcl-2, Bcl-XL, Bcl-W) are critical for cell survival, since loss of any of them causes cell death in certain cell type. However, its role during early porcine embryonic development is not sufficient. In this study, we traced the effects of Bcl-2 inhibitor, ABT-737, on early porcine embryonic development. We also investigated several indicators of developmental potential, including gene expression (apoptosis-related genes) and apoptosis, which are affected by ABT-737. Porcine embryos were cultured in the PZM-3 medium with or without ABT-737 for 6 days. In result, significant differences in developmental potential were detected between the embryos that were cultured with or without ABT-737 (14.7±3.0 vs 30.3±4.8%, p<0.05). TUNEL assay showed that the number of containing fragmented DNA at the blastocyst stage increased in the ABT-737 treated group compared with control (4.7 vs 3.7, p<0.05). The mRNA expression of the pro-apoptotic gene Bax increased in ABT-737 treated group (p<0.05), whereas expressions of the anti-apoptotic Bcl-2 family members (Bcl-2, Bcl-XL, Bcl-W) decreased (p<0.05). Also, expressions of the ER stress indicator genes (GRP78, XBP-1 and sXBP-1) increased in ABT-737 treated group (p<0.05). In conclusion, Bcl-2 is closely associated with of apoptosis- and ER stress-related genes expressions and developmental potential in pig embryos.

This study was carried out to investigate effective condition for producing somatic cell nuclear transfer (SCNT) embryos of Jeju native cattle. As donor cells for SCNT, ear skin cells from Jeju native cattle were used. In experiment 1, the effect of recipient oocyte sources on the development of Jeju native cattle SCNT embryos were examined. Fusion rate of recipient oocyte and donor cell was not different between the Hanwoo and Holstein recipient oocytes (86.0% vs 89.9%). The rate of embryos developing to the blastocyst stage was significantly (p<0.05) higher in Hanwoo recipient oocytes than in Holstein recipient ones (28.2% vs 14.7%). Blastocysts derived from Hanwoo recipient oocytes contained higher numbers of total cells than those derived from Holstein ones ( vs ), although there were no significant difference. The mean proportion of apoptotic cells in blastocyst was not different between the sources of recipient oocytes. In experiment 2, the development of Jeju native cattle and Hanwoo SCNT embryos were compared. Hanwoo oocytes were used as the recipient oocytes. Fusion rate was not different between the Jeju native cattle and Hanwoo SCNT embryos (92.1% vs 92.9%). The blastocyst rate of SCNT embryos was significantly (p<0.05) lower in Jeju native cattle than in Hanwoo (16.9% vs 31.0%). Blastocysts derived from Jeju native cattle SCNT embryos contained smaller numbers of total cells than those derived from Hanwoo ones ( vs ), but there were no significant difference. The mean proportion of apoptotic cells in blastocyst was not different between the Jeju native cattle and Hanwoo SCNT embryos. The present study demonstrated that Hanwoo recipient oocytes were more effective in supporting production of Jeju native cattle SCNT embryos, although Jeju native cattle SCNT embryos showed reduced developmental capacity when compared to Hanwoo SCNT embryos.

Malignant tumor of the paranasal sinus is a rare, occurring most frequently in the maxillary sinus. Carcinomas of the maxillary sinus are usually diagnosed at the advanced stage because most tumors have no symptom or nonspecific symptoms such as pain, nasal obstruction, rhinorrhea, and epistaxis. In addition to these features, it is difficult to distinguish carcinoma from inflammatory or cystic lesion on imaging study until the carcinoma destroys the surrounding structures. Therefore, the diagnosis is prone to be delayed. Here, we report a case of an 83‐year‐old male with nonkeratinizing carcinoma on the maxillary sinus, which was initially misdiagnosed as a cystic lesion. The aim of this study is to emphasize the effort for early diagnosis in order to improve prognosis and avoid inadequate treatment

The extracellular polysaccharide production from suspension culture of Aloe vera L. was analyzed, and the optimization for suspension cultivation was investigated. From specific polysaccharide assay of glucomannan, FT-IR and 1H NMR spectrum, the extracellular polysacchride from suspension culture was found to be similar to β-1,4-glucomannan originated from Aloe vera fresh leaf. The polysaccharide existed in an acetylated form and its molecular weight by gel permeation chromatography was estimated to be 490 kD (Mn = 440 kD; polydispersity = 1.115). From medium optimization by simplex-cemtroid design using MINITAB®, the callus growth on the 2,4-D of 11.82 μM, kinetin of 13.84 μM and α-naphthalene acetic acid of 4.34 μM was turned out to be significantly superior than the growth on the other culture media combinations. Under this optimal culture media, other conditions such as the suspension culture's temperature, pH and the growth terms were determined. These results showed that the callus growth was optimum at 25oC, pH 5.5 and in 2 weeks. The maximum levels of growth and extracellular polysaccharide production of callus by optimal conditions were about 20.4 and 2.5 g/L, respectively, showing the profile of extracellular polysaccharide production was closely related to the callus growth.

In this study, we determined total polyphenol content(TPC) and total flavonoid content(TFC) of extracts from Korean cabbage and cabbage using a spectrophotometric method as well as glucosinolates concentration by HPLC. TPCs of Korean cabbage and cabbage extracts were 308.48 ㎍ GAE/g dry weight and 344.75 ㎍ GAE/g dry weight, respectively. TFCs of Korean cabbage and cabbage extracts were 5.33 ㎍ QE/g dry weight and 5.95 ㎍ QE/g dry weight, respectively. We found six different glucosinolates, namely progoitrin, glucoalyssin, gluconapin, glucobrassicanapin, glucobrassicin and 4- methoxyglucobrassicin in the Korean cabbage extract. In the cabbage extract, there was four glucosinolates, namely glucoraphanin, sinigrin, glucobrassicin and 4-methoxyglucobrassicin. We determined the cytotoxic effect of Korean cabbage and cabbage extracts in AGS human stomach cancer cells, HepG2 human hepatic cancer cells and LNCaP human prostate cancer cells by MTT assay. Dose-dependent relationships were found between the extract concentrations and cancer cell growth inhibition. The overall results support that both Korean cabbage and cabbage, the major vegetables in Korea, contain bioactive compounds such as polypheol, flavonoids as well as glucosinolates and they may play a positive role in cancer prevention.

Several recent studies have detected genetic and cytogenetic alterations in epithelial odontogenic tumors. However, the detailed mechanisms of oncogenesis, cytodifferention, and tumor progression remain unknown. p63 as p53 homolog gene has been identified at loci 3q27-29. The p53 signaling cascade has an important role in oncogenesis or cyto- differentiation of odontogenic epithelium. Recently, several syndromes associated with p63 gene mutations have shown to include various tooth abnormalities of both the primary and permanent dentition. But little is known about p63 expression in odontogenic tumors, especially ameloblastomas. The purpose of this study were to examine various expression of p63 in ameloblastomas by immunohistochemistry and to clarify the possible biological role of p63 in ameloblastomas. 15 specimens including 6 follicular, 4 plexiform, 3 acanthomatous, and 2 granular cell types were fixed in 10% neutral formalin. 4um thick sections were used for routine H&E and immunohistochemical examinations. After immuno- histochemical satining, they were examined at a final magnification of 400X. For each case a minimum of 1000 nuclei located in the central and peripheral layers were counted in up to 10 consecutive microscopic fields per case. The immunoreactive cells were evaluated semiquantitatively. Immunoreactivity for p63 in all the types of ameloblastomas was higher in peripheral neoplastic cells than in central neoplastic cells. Keratinizing cells in acanthomatous ameloblastoma and granular cells in granular cell ameloblastoma showed markedly decreased reactivity for p63 in acanthomatous and granular cell ameloblastoma. Labelling index of acanthomatous, plexiform, and granular cell type was 86±11%, 81±17% and 83±15% in peripheral area while 88±14%, 82±11% and 76±10% in central area, respectively. Labelling index of follicular type was 17±4% in peripheral area while 21±3% in central area. There was no significant relationship between plexiform, acanthomaous, and granular cell type, while significant relationships between follicular and acanthomatous type, between plexiform and follicular type, and between granular cell and follicular type, respectively. It suggested that p63 expression could paly an important role in the pathogenesis of ameloblastomas. Morever plexiform, acanthomatous, and granular cell type would show more aggressive proliferative potentiality than follicular type.

Numerous reports on the immunopathogenesis of an oral lichen planus (OLP) have shown some inconsistency in terms of the role and distribution of CD4+ and CD8+ T cells, and Langerhans cells (LC). This has led to the hypothesis that although the clinical and histopathological manifestations of OLPs appear similar, they can be grouped into CD4+ predominant lesions and CD8+ ones on the varied immunopathogenesis or clinical progress. This study investigated the distribution of CD4+ T cells, CD8+ T cells, LC and the ratio of CD4/CD8 and defined the important immunocompetent cells. In addition, the clinicopathological and histopathological correlation with those immunocompetent cells were investigated. Frozen sections of 16 OLPs and 5 normal buccal mucosae were immunostained. Quantification was done using KAPPA Imagebase software and statistical analysis using SPSS 10.0 statistical package. CD8+ T cells were consistently more abundant in the epithelium of the OLP than CD4+ T cells, but no difference between the two cells was detected in the lamina propria. The intra-epithelial CD4+ T cells revealed a positive linear correlation with the intra-epithelial CD8+ T cells. In the lamina propria, the number of LC had a positive linear correlation with CD4/CD8 ratio. The number of LCs was higher in the reticular type of OLP compared to the erosive type. The histopathological features such as hyperkeratosis, acanthosis, the band-like infiltration of mononuclear cells, and liquefaction degeneration did not show significant correlation with the CD4/CD8 ratio. The results suggest that CD8+ T cells play major role in immunopathogenesis of OLP. The immunopathogenesis of OLP appears to vary from lesion to lesion in relation to the clinical progress.