Spermatogenesis is initiated from spermatogonial stem cells (SSCs) that has an ability of self-renewal and unipotency to generate differentiating germ cells. The objective of this study is to develop the simple method for derivation of SSCs using non-sorting of both spermatogonia and feeder cells. Simply uncapsulated mouse testes were treated with enzymes followed by surgical mincing, and single cells were cultured in stempro-34TM cell culture media at 37℃. After 5 days of culture, aciniform of SSC colony was observed, and showed a strong alkaline phosphatase activity. Molecular characterization of mouse SSCs showed that most of the mouse SSC markers such as integrin α6 and β1, CD9 and Stra8. In addition, pluripotency embryonic stem cell (ESC) marker Oct4 were expressed, however Sox2 expression was lowered. Interestingly, expression of SSC markers such as Vasa, Dazl and PLZF were stronger than mouse ESC (mESC). This data suggest that generated mouse SSCs (mSSCs) in this study has at least similar biomarkers expression to mESC and mSSCs derived from other study. Immunocytochemistry using whole mSSC colony also confirmed that mSSCs generated from this study expressed SSC specific biomarkers such as c-kit, Thy1, Vasa and Dazl. In conclusion, mSSCs from 5 days old mouse testes were successfully established without sorting of spermatogonia, and this cells expressed both mESC and SSC specific biomarkers. This simple derivation method for mSSCs may facilitate the study of spermatogenesis.

A Warthin’s tumor of major salivary glands, in particular of parotid glands, is a common benign tumor that may occur synchronously or metachronously in the same or contralateral gland. Moreover, epithelial malignance associated with a Warthin’s tumor is extremely rare, and exists in three forms; epidermoid carcinoma, adenocarcinoma, and undifferentiated carcinoma. The reports, related with a case of squamous cell carcinoma arising in a Warthin’s tumor at the parotid gland were reported only additional 3 cases from 1999 to 2010; 30 cases reported up to 1999.[2,4,7] This case report was a extremely rare case where both a primary squamous cell carcinoma and a Warthin's tumor were coexisting in the same

Inflammation functions as a double-edged sword against external stimulus. For instance, inflammation can have anti-cancer effect and simultaneously can play cancer-promoting factors. Recent studies have shown that cytokine plays an important role in tumor biology by influencing tumor growth, invasion and metastasis. We classify these cytokines by cancer type and review current knowledge of cytokines in terms of carcinogenesis. Here, we also focus on whether cytokines can act as biomarkers for early detection of oral squamous cell carcinoma (OSCC). This review will provide basis for further approach to study the role of cytokines in carcinogenesis and evaluating the possibilities of cytokines as biomarkers for cancer detection

Osteoblastoma is a very rare disease and in case of the jaw, maxilla infrequently occurs more than mandible. On account of difficulty to discriminate from other benign tumor or sarcoma, we need to pay attention to make differential diagnosis based on clinical, radiographic, pathological findings. A tumor on the maxilla and maxillary sinus could lead to a bad result for aesthetic appreciation and function. Therefore, reconstruction after maxillectomy is one of the most important issues concerning a surgery. The aim of this study is to report a case of osteoblastoma on the left maxilla which was misdiagnosed as ossifying fibroma and was treated with hemimaxillectomy and reconstruction using deep iliac circumflex artery musculo-osseous flap which resulted in satisfactory result.

New therapeutic measure are needed to improve the outcome for patients with oral squamous cell carcinoma(OSCC) because OSCC continues to portend a relatively unfavorable prognosis. Recently RNA interference(RNAi) has emerged as an effective method to target specific genes for silencing. Although overexpression of urokinase-type plasminogen activator receptor(uPAR) has been implicated in progression and metastasis of OSCC, the transfection effect of RNAi- uPAR on OSCC has been rarely reported. The purpose of this study were to examine the efficient and specific inhibition of uPAR mRNA and protein expression by siRNA targeting of uPAR through RT-PCR and immunoslot blotting, and to study cell proliferation activity, adhesion, invasion and migration in vitro compared to the controls. In MTT assay, siRNA-uPAR transfected cells showed about 70-80% cell proliferation compared to OSCC cell lines after 2 days. In adhesion assay, siRNA-uPAR transfected cells showed about 20-30% adhesion activity compared to OSCC cell lines, but similar features to those of BSA coated wells. In migration assay, siRNA-uPAR transfected cells showed about 60% migration activity compared to OSCC cell lines, but higher 3.5 folds to those of BSA coated wells. In invasion assay, siRNA-uPAR transfected cells showed about 55% invasive activity compared to parental cell lines. mRNA expression of siRNA-uPAR transfected cells showed about 10-15 % compared to parental cell lines by RT-PCR. Protein expression of siRNA-uPAR transfected cells showed about 25% compared to parental cell lines by ELISA assay. It suggested that RNAi-uPAR tranfection might be used as a potent and specific therapeutic tool for the treatment of oral squamous cell carcinoma, especially in inhibiting invasion and metastasis.

지구 온난화의 영향으로 우리나라는 지난 30년동안 평균기온이 0.7℃, 겨울철에는 1.4℃가 상승하였다. 이러한 온난화로 인하여 우리나라에서는 이상기상 현상이 자주 발생하여 채소작물에 피해가 발생한다. 특히 노지에서 많이 재배되고 있는 고추, 배추 및 무는 온난화로 인하여 정식시기를 점점 앞당겨 정식후 갑작스런 저온이 오면 이들 작물의 피해가 크다. 따라서 본 실험은 저온에 따른 배추의 생육특성과 엽 세포조직에 미치는 영향을 구명하고자 실시하였다. '춘광' 배추품종을 화분에 정식한 후 노지 처리구, 무가온 하우스 및 가온하우스 처리구 등 3처리를 하였다. 그 결과, 정식후 50일의 생육은 노지처리구의 초장, 엽수, 엽록소 및 엽면적이 가온 처리구에 비해서 현저하게 떨어졌고, 특히 생체중의 경우에는 가온 처리구에 비해서 노지와 무가온 하우스 처리구가 1/3 수준으로 현저하게 낮았다. 배추의 잎이 10매 정도 생육이 되었을 때 저온에 따른 배추 잎의 피해증상은 영하 3.0℃ 조건에서는 배추 겉잎에 약간의 수침증상을 보였으나 회복되었다. 그러나 영하 7.4℃ 조건의 배추 잎은 수침증상이 심하였으며 회복되지 못하고 황색으로 변하면서 결국 잎이 고사하였다. 피해받은 잎의 엽육세포는 영하 3.0℃ 조건에서는 울타리조직과 해면조직이 약한 붕괴증상을 보였지만 어느정도 형태를 갖추고 있었는데, 영하 7.4℃ 조건에서는 세포가 동결된 후 해동되는 과정에서 세포의 막구조가 파괴되어 울타리조직과 해면조직이 완전히 붕괴되었기 때문에 세포 형태를 갖추고 있지 않았다. 따라서 배추 정식후 초기 생육 단계에서 영하 3℃까지는 비닐이나 부직포로 보온, 토양수분 조절, ABA 처리를 하여 동해를 예방할 수 있으나 영하 7℃의 저온이 발생하면 세포가 파괴되어 회복하기 어렵기 때문에 다시 심거나 또는 다른 작물로 대체하는 것이 좋을 것으로 사료되었다.

Abstract. Inflammatory myofibroblastic tumor (IMT) is considered as a benign tumor with local aggressive course, consisting of myofibroblastic spindle cells with an inflammatory cells infiltration. IMTs are more usually found in the lung and very rarely in the mandible. We report an IMT of the mandible in a 54-year-old man. The patient complained of pain on the right side of mandible. Radiographically, the lesion was occupied in the right mandible with bone destruction. Although the initial diagnosis was an osteomyelitis, the histopathologic examination and immunohistochemistry revealed it to be an IMT. Histologically, the lesion was composed of inflammatory cells infiltration within a variably fibroblastic or myofibroblastic spindle cell background. Immunohistochemically, spindle cells stained with smooth muscle actin (SMA) and CD68 (KP1), but uniformly negative with desmin and cytokeratin.

Small cell osteosarcoma of bone, which was first described in 1979, is an unusual variant of osteosarcoma. Osteoid production by tumor cells is frequently focal or minimal, making the differential diagnosis with other small round cell tumors of bone difficult. Here, we present a rare case of small cell osteosarcoma of the mandible appearing as bony bulging mass in 31-year-old male who has neither tenderness nor paresthesia. Histologically, the tumor contains hypercellular cartilage and abnormal osteoid associated with small round to ovoid malignant cells. Awareness of small cell osteosarcoma should be emphasized because it has worse prognosis than both other small round cell tumor and conventional osteosarcoma.

Neurotized melanocytic nevus (NMN) is categorized into intradermal/intramucosal type of acquired melanocytic nevus. In contrast to typical intramucosal nevus which has relatively distinct histological features, the diagnosis of NMN requires more attention due to its mimicry of benign neural tumors such as neurofibroma. The majority of lesional cells, NMN cells, showed a spindle cell morphology and abundant, eosinophilic cytoplasm which were positive for S-100, vimentin, and collagen type IV. Positive reaction for MART-1 was detected in the NMN cells as well as in the epithelioid nevus cells beneath the epithelium. Neurofibroma exhibited diffuse positivity for S-100, vimentin, CD34 and collagen type IV, but never expressed MART-1. Toluidine blue stained the numerous mast cells scattered in the lesion of neurofibroma, compared to the relatively minor detection of mast cells in NMN. Therefore, MART-1 is a useful marker in differentiating NMN from neurofibroma.

Oral squamous cell carcinoma (OSCC) is one of the most common carcinomas in the head and neck area. Bitter melon extraction (Momordicacharantia, BME) has been used as a functional food to prevent and treat human health related issues. It has recently been reported that BME inhibits breast cancer cell growth by arresting cell cycle and promoting apoptosis. In this study, we aimed at proving the inhibitory action of BME on OSCC proliferation. We used two OSCC cell lines, SCC4 and Ca9-22. Both cell lines were treated with different concentrations (1%, 2%, and 5%) of BME. Cell viability was examined by MTT assay. DNA condensation was visualized by immunofluorescence microscope to determine the signs of the cell apoptosis. Cell numbers were significantly decreased in a dose-dependent manner by bitter melon at concentration of 1% of BME on Ca9-22 cell line (P=0.001) but no significant effect on SCC4 (P=0.124) at the same concentration. 2% BME treatment of the Ca9-22 cell line induces chromatin condensation and DNA fragmentation after 20 hours. It seems that BME inhibits the proliferation of Ca9-22 cell line by inducing apoptosis. Thus, BME may be used as a dietary supplement for prevention of OSCC.

Odontoblasts and/or dental pulp cells are responsible for tooth repair as well as dentin formation. Adhesion and migration are critical processes for tissue regeneration. This study was performed to clarify whether Pam3 modulates adhesion and migration of a murine odontoblast-like cell line, MDPC-23 cell and TLR2 signaling is engaged in this process. TLR2 expression in MDPC-23 cells was examined by RT-PCR. Adhesion assay was performed using type Ⅰ collagen-coated plates. Migration ability was determined by a commercial assay kit. Phosphorylation of IκB-α, JNK, p38, and ERK was examined by Western blot analysis. TLR2 was functionally expressed in MDPC-23 cells. Pam3CSK treatment enhanced adhesion and migration of MDPC-23 cells in a dose-dependent manner. Blockade of TLR2 using its antibody restored Pam3CSK-induced adhesion and migration of MDPC-23 cells. These findings indicate that Pam3CSK, an immune activator from gram negative bacteria, can promote adhesion and migration ability of MDPC-23 cells via TLR2.

본 연구는 복숭아 연화의 기작을 밝히기 위하여 ‘장호원황도’를 공시하여 과실품질 및 세포벽 관련물질의 변화를 조사하였다. 과실을 성숙기에 수확하여 과실 경도별 4그룹으로 구분한 후 25℃에서 10일간 보관하면서 품질특성을 조사하였다. 연화가 진행될수록 과육경도의 유의한 감소와 가용성고형물의 증가가 나타났다. 당의 구성은 서당의 비율이 가장 높았는데 연화초기에 84.0%, 연화말기에서 75.8%로 나타났고 연화과정이 진행되면서 환원당의 비율이 점진적으로 증가하는 것으로 조사되었다. 알콜불용성 물질 함량은 연화만기부터 감소하기 시작하였는데 수용성펙틴의 가용성도 함께 증가하기 시작하였다. 과육이 완전히 용질된 연화말기에는 수용성 및 CDTA 가용성펙틴과 4% KOH 가용성 헤미셀룰로스의 가용성이 크게 증가하였다. CDTA 가용성펙틴의 분해는 연화만기에 시작되어 연화말기에 극심하였고 4% KOH 가용성 헤미셀룰로스의 분해는 연화말기에만 나타났다. 종합적으로 볼 때 복숭아 ‘장호원황 도’의 연화과정은 펙틴과 4% KOH 가용성 헤미셀룰로스의 가용성 증대를 수반하며 셀룰로스에 약하게 결합되어 있는 matrix glycan의 부분적 분해와 관련이 있다고 보여진다.

Gingival fibroblasts (GF) are the most abundant cell type in periodontal connective tissues, andhave distinct functional activities in the repair of periodontal tissues and in inflammatory periodontal diseases. Human gingival fibroblasts (hGF) can be used for periodontal tissue engineering. This study examined whether the alkaline phosphatase of hGF is enhanced by recombinant human BMP-4 and/or Anti human BMP-4 antibody. hGF was obtained from the excised gingival tissue of an implant patient undergoing 2nd surgery. The tissue was incubated at 37℃ in 5% CO2 and 95% humidity, and the cultivating media was changed every 2 days. The 2nd passage hGF cells were cultured in a medium containing Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum and 1 X antibiotic antimycotic solution. The control hGF was cultured for 7 days without rhBMP-4/Anti human BMP-4 antibody. The experimental groups were cultured for 7 with BMP-4 (10 ng/ml) and/or Anti human BMP-4 antibody. This study evaluated the differentiation of hGF to osteoblasts using alkaline phosphatase assay. In the experimental groups, the hGF showed abundant positive ALP staining. Among the experimental groups, the experimental group 3 (mixture of rhBMP-4 (20ng/㎖) and Anti human BMP-4 antibody (50000ng/㎖) showed most abundant positive ALP staining. In the control group, the hGF showed weak positive ALP staining. Overall, these results suggest that the ALP expression of hGF can enhanced by rhBMP-4 or mixture of rhBMP-4/ anti human BMP-4 antibody.

This study was designed to investigate the effect of fucoidan on the activation of macrophage and on induction of apoptosis in AGS cell. To measure the activity of macrophages, NO and TNF-α assays were performed in Raw 264.7 cell. Treatment with fucoidan significantly increased production of NO and TNF-α, indicating activation of macrophages. The result of MTT assay shows that cell viability was significantly decreased in a dose and time-dependent manner. Fucoidan increased to enhance mitochondrial membrane permeability, as well as the cytochrome c release from the mitochondria. Fucoidan decreased Bcl-2 and XIAP expression, whereas the expression of Bax was increased in a time-dependent manner compared to the control. In addition, the active forms of caspase-9 were increased, and the inactivation of Akt was decreased in a time-dependent manner. Caspase inhibitor, z-VADFMK, canceled the apoptosis of fucoidan, expression of Bax and caspase-9 were decrease. These results indicate that fucoidan induces activation of macrophage and apoptosis through activation of caspase on AGS cell.

Ameloblastic fibro-odontoma has been defined as a lesion similar to ameloblastic fibroma by WHO, as it shows inductive changes which forms enamel and dentin. Ameloblastic fibro-odontoma is a very rare mixed dentition tumor in children, and the symptom shows indolent edema in maxillary and mandibular molar area. The prevalence is two times higher in male than in female, and two times higher in maxilla than in mandible. Radiologically, it shows clear border and characteristics of both fibroma and odontoma histologically. This review reports a case that a 4-year old female visited in dental clinic of this school for edema as chief complaint in Feb, 2012. Emergency surgical curettage was performed right after initial diagnosis as odontoma, then confirmed diagnosis as Ameloblastic fibro- odontoma after biopsy. Currently, after 6 month, no sign of recurrence can be seen. Ameloblastic fibro-odontoma is very rare mixed dentition tumor. Moreover, as it is the case of female maxilla, this case is worth of publishing. Furthermore, accurate diagnosis of Ameloblastic fibro-odontoma is difficult. This review is published for accurate diagnosis through differential diagnosis of several important mixed dentition tumors.

Rushton bodies are known to be the aberrant keratinization and calcification in the epithelium of odontogenic cyst, which are similar to the features of calcifying odontogenic cyst and pilomatricoma. However, the pathogenetic mechanism of keratinization and calcification of Rushton bodies has not been clearly elucidated. Here, a case of Rushton bodies found in dentigerous cyst was examined by immunohistochemical method using antisera of PCNA, pAKT, HIF, PIM1, and PARP. The globular keratinization in lamellate fashion showed weak birefringency under polarizing microscope, and the Rushton bodies frequently underwent the dystrophic calcification. The polygonal keratinocytes of Rushton bodies were strongly positive for HIF and PARP, and the cyst epithelium was diffusely positive for pAKT and PIM1. Particularly, the cyst epithelium was hyperplastic and focally invaginated into cyst wall with positive reaction of PCNA. These findings may indicate the active response of odontogenic epithelium against the apoptotic stress of the cyst, producing the globular keratinization and irregular calcification in the polygonal keratinocytes. Therefore, it is presumed that the lamellate keratinization and dystrophic calcification of Rushton bodies are aberrant products of retrograding keratinocytes slowly undergoing apoptotic progresses similar to the phenomena of the ghost cells in calcifying odontogenic cyst and pilomatricoma, and also may have a potential for oncogenic proliferation.

Internalization and expression of extracellular molecules into cells and tissues is known very important process to biological processes and therapy of various diseases. In this study, we analyzed expression pattern of extracellular molecule after transduction into various human cells. To investigate cellular expression of internalized molecule, we used adenovirus containing green fluorescence protein. After infection of adenovirus into various human cells, the efficiency of intracellular gene expression was assessed with determining GFP expressing cells by fluorescence microscopy or FACS. After one day of adenovirus infection into HepG2 and A549, we observed that GFP expression was low at 10moi but expression levels were increased at 100moi in both cells. But, adenovirus infection into HCT116 showed low expression of GFP at concentrations from 1moi to 100moi. After 2 day infection with adenovirus, GFP expression level at 10moi and 100moi was highly increased in HepG2 and A549 compared with 1 day infection. Especially, GFP expression was significantly increased in HCT116 after 2 days infection. However, GFP expressing SKOV3 cells by adenovirus infection were not found in all the experimental conditions tested. For quantitative analysis of GFP expression of cells by adenovirus infection, we carried out FACS analysis. As a result, GFP was expressed at very low levels at 1moi in all cells used in this experiment. GFP expression slightly increased after increasing moi to 10 in HepG2, HCT116, and A549 cells. By 100moi infection of adenovirus, GFP expression was elevated to 10 fold higher than 10moi in HepG2 and A549 and about 4 fold elevation was observed in HCT116. A549 showed 20 fold higher expression of GFP than SKOV3. We also found that GFP expression by adenovirus infection was the highest in HepG2 cells. Protein expression was enhanced by increasing concentrations or time of adenovirus infection. In these results, GFP expression efficiency of adenoviral gene transduction reveals the highest in HepG2 and lowest in SKOV3 among the cells tested. Taken together, we could confirm that intracellular protein expression efficiency by transduction of extracellular gene was different in various human cells. Our study suggests that the cell types and cellular properties should be carefully examined to enhance expression efficiency of extracelluar molecules in biological research and disease therapy

Cellular microenvironment is an essential issue for regulating epithelial characteristics through the alteration of intricate signaling pathways and intercellular communications in different cell types. Thus, microenvironment influences tumor initiation, progression, and metastasis. This study aimed to investigate the relationship between microenvironment and epithelial property in HPV16 E6/E7-immortalized human oral keratinocytes (IHOKs). To investigate characteristics of IHOK cultured in different media, two media were used, which included keratinocyte growth media (KGM), F-medium composed of 3:1 ratio of DMEM and F-12 (P media) supplemented with 10% FBS and 1% penicillin/streptomycin. Proliferative property and invasive and migratory activity were observed. As results, proliferating activities of IHOK in different culture condition were changed. Likewise, migratory and invasive activities were also different depending on media types. These results suggest that cellular microenvironment can affect modification of biological properties of epithelial cells.

Considering the dearth of information regarding the medicinal properties of Luffa cylindrica, we assessed the antioxidative, antimutagenic and hyperplasia inhibitory activity of cancer cells from Luffa cylindrica extracts by employing biological and biochemical assays. Ethanol extracts of Luffa cylindrica inhibited MDA-BSA (malondialdehyde-bovine serum albumin) conjugation reaction (66.38±2.65), DPPH (1, 1-diphenyl-2-picryl-hydrazyl) radical production (60.13±0.42) and lipid peroxidation (56.04±3.24). In this study, Luffa cylindrica is believed to exert possible antioxidative effects. The direct and indirect antimutagenic effects of the ethanol extracts of Luffa cylindrica were examined by the Ames test using Salmonella typimurium TA98 and TA100. The inhibitory effects on indirect and direct mutagenicity shows an weak tendency, particularly in direct mutagenicity mediated by 2-nitrofluorene in Salmonella typimurium TA98 (5.82±5.74) and in indirect mutagenicity mediated by 2-anthramine in Salmonella typimurium TA100 (5.76±2.15). The ethanol extracts of Luffa cylindrica on cancer cell hyperplasia inhibitory activity via MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyltetrazolium bromide) assay exerted cytotoxic effects on Hela cells (55.83±3.83) and MCF-7 cells (33.03±2.09), which were used in this study. Based on these results, it believed that the ethanol extracts of Luffa cylindrica have antioxidative capacities as well as hyperplasia inhibitory activity of cancer cells. Furthemore, Luffa cylindrica is a candidate for the prevention and dietetic treatment of chronic diseases and for the development of functional food.