All organisms are being exposed to harmful factors present in the environmental. The combined action of various factors is a distinguishing feature of modern life. An interaction between two chemicals is considered as synergistic when the effect produced

Elevated expression of survivin is strongly associated with tumorigenesis and even in human common cancers. Oral squamous cell carcinoma (OSCC) is the 7th most frequent cancer in human and responsible for more than 90% of all oral cancer. The purpose of this study is to confirm whether survivin is associated with oral carcinogenesis, expecially has a role in the development of OSCC. For the control group; 3 specimens obtained from normal oral mucosa without any inflammatory reaction were used a nd for the experimental group, specimens obtained f rom 18 sub jects of OSCC; 6 subjects from Well differentiated type OSCC; 4 subjects from Moderately differentiated type OSCC; 3 subjects from Poorly differentiated type OSCC; 3 subjects from Verrucous carcinoma: and 2 subjects from C arcinoma in situ were used. All the specimens were embedded in paraffin, sectioned 5 μm or more in thickness, and stained with hematoxylin- eosin. For immunostain, the specimens were incubated with 1;200 diluted primary antibody (anti-survivin monoclonal, Biocare Inc, USA), followed by the secondary antibody(NovoLink Polymer detection system, Novocastra Lab., UK). The bound antibodies were visualized by addition of diaminobenzidine tetrahydrochloride(DAB) for 30 minutes at room temperature. The specimens were counterstained with Mayerʼs Hematoxylin and mounted. Quantitation of immunoreactivity was performed under the light microscope with the following criteria ; Intensive reaction; +++, Moderate reaction; ++, Minimal reaction; +. Using the image analyzer(Korea Optical System), immunoreactivity of tumor cells in various field was measured and statistically analyzed with SPSS 15.0 Program. The results were as follows: Expression of survivin in OSCC was significantly increased in the nucleus and the cytoplasm of OSCC as compare to those of control group (p<0.05). Expression of survivin in the nucleus and the cytoplasm of the cells in OSCC is correlated with the cellular malignancy (p<0.05). Expression of survivin in Poorly differentiated type OSCC partly correlated to some extent to cellular malignancy (p<0.05). These results suggest that expression of survivin in OSCC is closely associated with to the development, and malignancy of the OSCC, b ut it is not enough to be used a s a marker f or the c ellular malignancy. Further studies are needed to relate the expression of survivin to cellular malignancy.

Artemisia scoparia (A. scoparia), perennial herb is indigenous to Korea and has been traditionally used in liver damage. We investigated the effect of the essential oil obtained from A. scoparia on apoptosis of KB cells. Cytotoxicity and cellular DNA content were analyzed by MTT assay and flow cytometry, agarose gel electrophoresis, and Hoechst 33258 staining. The caspase-3 and poly (ADP-ribose) polymerase (PARP) proteins were estimated by Western blotting method. We found that the essential oil induced the apoptosis of the KB cells by concentrations of 0.4 to 0.2 mg/ml which was verified by DNA fragmentation, apoptotic bodies, and the sub-G0/G1 ratio. The essential oil also transient caspase-9 and caspase-3 activity and cleavage of PARP in KB cells for 24 h. The essential oil-induced apoptotic cell death was accompanied by up-regulation of Bax and down-regulation of Bcl-2. In conclusion, we demonstrated that the essential oil of A. scoparia induces apoptosis in KB cells

Tumor cells under hypoxic conditions are often found due to the rapid outgrowth of their vascular supply, and,in order to survive hypoxia, these cells induce numerous signaling factors. Erk is an important kinase in cell survival, and its activity is regulated by Raf kinases through numerous growth factor receptors. The authors investigated Erk activation and Raf/Erk signaling using the hypoxia-mimetic agent, cobalt chloride (CoCl2), in oral squamous cell carcinoma (OSCC) cells. CoCl2 increases Erk phosphorylation in both a dose- and time-dependent manner. In addition, blocking the activation of epidermal growth factor receptor (EGFR) using PD168393 abolished Erk activation in response to CoCl2, suggesting that Erk phosphorylation by CoCl2 is dependent on EGFR.

This study was performed to investigate the effect of extract mixture(IPGE) drink from Inonotus Obliquus, Phellinus Linteus and Ganoderma Lucidum on hematopoietic stem cells and lymphocyte subset[lymphocyte, CD4+ T cell, CD8+ T cell, Natural Killer(NK) cells] of blood in 37 participants who were healthy and about 40~70 years old. They were divided into two groups; extract mixture drink administration group(n=27) and placebo administration group(n=12). They were given the test drink daily for 4 weeks. Blood was obtained from the subjects every two week in the beginning of administration day to evaluate the CD34+ hematopoietic stem cells and immune cells. As results, CD34+ hematopoietic stem cells were significanly increased after taking IPGE drink for 4 weeks compared to that before taking the drink (p < 0.001). There was no significant changes in number of lymphocytes, CD4+ T cells, CD8+ T cells, NK cells and in the ratio of CD4+/CD8+ cell after taking the test drink. From these results, it was suggested that IPGE have a good health effect by promoting the proliferation of the hematopoietic stem cells.

The differentiation of leukemia cells into mature cells is a major target of the human leukemia therapy. As differentiated leukemia cells lose their proliferative and tumor-forming abilities, differentiation inducers may be useful for the treatment of leukemia. In this study, the experiments were designed to determine whether diallyl disulfide (DADS) regulates expressions of tumor suppressor protein PTEN (phosphatase and tension homologue) in HL- 60 cells. DADS causes upregulation of PTEN in a time- and dose-dependent manner, which was correlated with decrease of phospho-Akt level. These results suggest that DADS induces upregulation of PTEN in human leukemia cells. These results suggest that DADS may be a useful anticancer agent for management of human leukemia.

안토시아닌(Anthocyanin)은 플라보노이드계 화합물의 한 부류로 항산화, 항암 및 항궤양, 항당뇨, 중금속해독, 시력보호, 콜레스테를 저하 등의 다양한 생리활성을 가지는 것으로 보고되어 있다. 죽상경화과정은 염증성 사이토카인의 분비 또는 혈관손상으로 인한 백혈구의 부착과 이동을 통해 시작된다. 본 연구는 이러한 죽상경화의 초기과정에서 안토시아닌 혼합물 중 single compound인 delphinidin chloride (DC) 인간혈관 내피세포

곰취(Ligularia fischeri)의 이용성을 높이데 필요한 기초자료를 확보하고자 메탄올을 용매로 하여 추출 온도 및 시간에 따른 생리활성 효과를 조사하였다. 곰취 메탄올추출물 1,000mg·L-1에 함유된 총 페놀함량, 총플라보노이드 함량은 각각 75.8-297.7mg·L-1 and 45.6-173.6mg·L-1이었다. 추출온도와 시간은 총 페놀함량, 총 플라보노이드 함량, 전자공여 능 의 경우 95℃에서 6시간 동안 추출했을 때 가장 좋았다. 그러나 아질산염 소거능은 75℃에서 12시간 추출한 것에서 97.4%로 가장 높게 나타났다. 곰취 메탄을 추출물 200mg·L-1 및 400mg·L-1 농도는 각각 폐암과 위암세포 증식을 90% 이상 억제시켰다. 따라서 곰취는 높은 생리활성 기능을 나타내는 채소로 나타났으며, 곰취를 가공품으로 이용하기 위해 메탄올로 추출할 때는 95℃에서 6시간 정도 하는 것이 좋을 것으로 생각되었다.

당뇨망막병증은 서구에서 성인들의 실명을 일으키는 원인이다. 당뇨병이 있을 때 고혈당증은 여러 세포 형태에서 세포자연사를 유도하지만 그 기작은 명확하게 밝혀지지 않았다. 본 연구의 목적은 인간망막 내피세포에서 고혈당 포도당이 세포자연사에 미치는 영향에 대하여 알아보았다. 망막 내피세포는 5, 25, 50 mM 포도당이 포함된 IMDM배지에서 37℃, 5% CO2조정된 항온기에서 24, 36, 48시간 동안 배양하였다. 여러 농도의

본 연구는 한우의 난포낭종 및 황체낭종에서 세포자멸사 관련 유전자의 발현 변화를 조사함으로써 난소낭종과 세포자 멸사간의 관계를 정리하고자 한다. 마이크로어레이 분석 결과, 난포낭종에서 PIK3R2와 AKT1가 유의하게 증가하였고, 황체낭종에서는 증가되는 세포자멸사 관련 DEGs, TNF-RAF2, PRLR, FOXL2, STK4 및 COL4A3와 감소하는 DEGs, INHA, CIDEB, BCL10 및 FASLG가 포함되었다. 정량적 역전사 중합 효소

A 14-year-old intact female, mixed dog was presented with hematuria and strangury. Mass in the abdominal cavity was seen on radiographs and ultrasound. On the cytological examination in the urethra, clusters of pleomorphic epithelial cell were found. Tissues of the urethra and the urinary bladder were obtained at the time laparotomy and determined the extent of the mass. Transitional cell carcinoma (TCC) of the urinary bladder was found in histopathologic characteristics. Urinary diversion after removal of a complete full section of the TCC in bladder wall was performed. Piroxicam, as a medical therapy for TCC, was orally administrated. Surgical operation and chemotherapy were selected with the goal of maintaining and improving quality of life.

Epithelial-mesenchymal transition (EMT) can play an important role in carcinogenesis of oral squamous cell carcinoma (OSCC). EMT is characterized by morphological and phenotypical change of epithelial cells into mesenchymal cells, and transcriptional repressor of E-cadherin, Snail is critical for EMT. In order to investigate the role of Snail and E-cadherin in OSCC, we analyzed the immunohistochemical pattern of Snail and E-cadherin in 18 OSCCs. The expression of Snail in the OSCC was increased whereas the expression of E-cadherin in the OSCC was decreased in comparison with those of normal oral mucosa, showing reverse correlation. Especially, the fibroblasts near the islands of OSCC showed the positivity of Snail, suggesting the reactive fibroblasts to the EMT of epithelial tumor cells. In metastatic squamous cell carcinoma in cervical lymph node, the positivity of Snail of tumor cells was higher than that of primary OSCC. We concluded that the increased Snail expression and the decreased E-cadherin expression were involved in the progression, invasion and metastasis of OSCC.

Intracellular reactive oxygen species(ROS) produced in a various pathologic state was known to intermediate many cellular response such as inflammation. Recently, low level light irradiation by HeNe laser used in many clinical field could improve inflammatory state by scavenging intracellular ROS through photo-detachment/dissociation process. The purpose of this study is to investigate the differential effects of blue and red light irradiation on ROS scavenging effects. Immortalized human oral keratinocyte HaCat cells were used. Phorbol 12-myristate 13-acetate(PMA) was treated for inflammation. Red(635nm) and blue(470nm) light irradiation was carried out. To asses the intracellular ROS by light irradiation, confocal microscopic and flow cytometric assay with DCF fluorescence for total ROS and ESR spectrometry of DMPO-O2 - for superoxide anion were caried out. And microarray was performed for mRNA expression level. Released intracellular total ROS in PMA treated HaCat cell lines was dissociated efficiently by red light irradiation, while blue light irradiation did not. Rather, blue light irradiation increased ROS formation. For superoxide anion generated the first synthetic form of ROS, red light irradiation reduced its amount but blue light irradiation did not. In the mRNA expression in line with cyclooxygenase(COX) pathway, prostagrandin endoperoxide synthase 1(PTGS 1), prostagrandin endoperoxide synthase 2(PTGS 2) and phospholipase A2(PLA2) were increased by both light irradiation and they were decreased as time flows. And genes associated with ROS releasing, mRNA expressions of tumor necrosis factor receptor (TNFR) and interleukin 1beta(IL1B) were increased by 1 hour red light irradiation but did not by blue light irradiation. As a result, red and blue light irradiation showed different response in affecting the level of ROS. These findings indicate that red light rather than blue light is more useful for anti-inflammation in clinical field

The phytochemicals of many plants suggest their potential use as dietary supplements in cancer chemoprevention and chemotherapy. In the present study, antitumor activity of Cudrania tricuspidata, a plant native to East Asia, was investigated. Cell growth inhibition of the extract on HT-29 colorectal adenocarcinoma using MTT colorimetric assay was determined. Apoptosis on HT-29 cells was performed by DNA fragmentation analysis. PGE2 release was measured by enzyme immunoassay, because PGE2 is a key protumorigenic prostanoid in many human cancers. For the ROS scavenging activity, ROS level was detected by laser scanning confocal microscope. It was found that methanol extract of leaves inhibits cell viability by inducing apoptosis as evidenced by DNA fragmentation. Stem bark decreases synthesis of PGE2, inflammatory mediator. Fruits exhibited pronounced ROS scavenging activity. Taken together, these results suggest that Cudrania tricuspidata exerts growth inhibition and anti-oxidation on HT-29 cells through apoptosis, ROS scavenging respectively that it may have anti-cancer properties.

It is not yet clear to know how normal human osteoblasts(NHost) from oral and maxillofacial area deposit, stabilize, and configure their extracellular matrix on dental biomaterial surfaces. Therefore it is necessary to design biomaterials with improved biocompatibility that will promote earlier bone differentiation and mineralization. There is now increasing evidence that TGase 2 may play a role in the initiation and regulation of the mineralization processes and serves to cross-link osteocalcin and osteopontin, which are predominant substrates for TGase 2 expressed during bone mineralization. But it is still unclear as to which TGase 2 is expressed by NHost in vitro bone formation. The purpose of this s tudy was t o determine the level of TGase 2 expression associated with collagen I , osteopontin and osteocalcin in NHost cell lines from oral and maxillofacial area in vitro. We will investigate whether TGase 2 has an active role in the biocompatibility of dental biomaterials during differentiation and mineralization of NHost. NHost cell lines were cultured under DMEM with 10% FBS at 37ﾟC and 5% CO2. Collagen quantification, mineralization and calcium assay, ALP activity assay, and RT-PCR analysis during bone differentiation and mineralization were done. ALP levels showed higher activity under AA+hGP t han under AA. I nhibition o f T Gase 2 by cystamine showed d ecreased Ca++ concentration, c ollagen I deposition and ALP level during 11 days of differentiation. TGase 2 mRNA expression of NHost was constant during mineralization stage. TGase 2 enzyme activity was increased during differentiation. Osteopontin mRNA expression during mineralization was higher than that of osteocalcin and collagen I . It suggested that TGase 2 associated with collagen I, osteocalcin and osteonectin might play a role in the differentiation of NHost from oral and maxillofacial area, but a little involved in mineralization.

Cementum is a hard connective tissue, produced by cementoblasts during tooth root formation, which provides for the attachment of the periodontal ligament to the roots and surrounding alveolar bone. Establishment of this attachment is an important event in the regeneration of lost periodontal tissues. We examined whether or not odontoblast conditioned media(CM) have a regulatory influence on the differentiation and mineralization of cementoblasts(murine cementoblastic cell line, OCCM-30) in vitro. To identify the effect of odontoblast conditioned media and dentin non collagenous proteins (dNCPs) on cementoblast differentiation and mineralization, we treated CM and dNCPs to cementoblast then differentiated the cells for 14 days. To evaluate the formation of mineralized nodules alizarin-red S staining was performed at 0,4,7 and 14 days. Expression of cementum matrix genes was measured by RT-PCR. Mineralization of cementoblasts was accelerated with CM from odontoblastic MDPC-23 and OD-11. The expression of BSP, ALP, and OC mRNA in cementoblastic OCCM-30 cells was facilitated by the MDPC-23 and OD-11 cells. The extracted dNCPs had little influence on the proliferation, cell cycle modification, and chemotaxis of OCCM-30 cells. Although the dNCPs did not exhibit chemotactic activities for cementoblasts, the dNCPs promoted the differentiation and mineralization of cementoblasts. In conclusion, the dentin matrix protein, or the secreted products of odontoblast, facilitates cementoblast differentiation and mineralization. This represents a new approach and suggests another avenue for cementum regeneration.

Considering the great potential of iron chelators at inhibiting the proliferation of tumor cells, in order to determine the molecular and biological basis for the effects of iron chelator in oral cancer, we investigated the effects of iron chelator, desferrioxamine (DFO), on the gene profiling analysis of immortalized human oral keratinocytes (IHOK), and oral cancer cells (HN12), using the cDNA microarray. We identified 46 clones cDNA exhibiting more than 2 fold overexpression in DFO treated IHOK and HN12 cells, and 94 cDNA reveal more than 2 fold down-regulated expression. Examination of gene expression that differs between DFO treated vs. control IHOK and HN12 cells apprear to be related to : cell cycle regulator, cell growth and apoptosis, signal transduction and stress. p21 for cell cell cycle factor was upregualted, and cyclin-cdk gene was decreased expression, so we observed cell cycle arrest in DFO treated IHOK and HN12 cells. In tumor growth, we have identified downregulation of hemidesmosomal protein (bullous pemphigoid antigen 1) and epiregulin expression in DFO treated IHOK and oral cancer cells. Signal transducers including mitogen-activated protein kinase-activated protein kinase 5, serine/thereonine kinase 6 were downregulated with DFO treated cells, suggesting the DFO regulates the p38 MAP kianse pathway in immortalized and maignant oral keratincytes. In conclusion, we have demonstrated the high-throughput utility of cDNA array hybridization in parallel to the gene expression analysis to identify genes that are expressed differentially in DFO treated with immortalized and malignant oral keratinocytes. The differentially expressed genes identified here should be informative in DFO-induced anti-cancer effects.

The purpose of this study was to evaluate the role of integrin α3 and integrin β1 in the ameloblastomas. For this study, 10 specimens diagnosed as amoblastomas referred to the Department of Oral Pathology, School of Dentistry, Kyung Hee University, and 5 specimens of normal oral mucosa without any inflammatory changes were used as experimental and control groups, respectively. The ameloblastomas devided into follicular type, plexiform type, acanthomatous type, and granular cell type. All specimens; experimental and control group were fixed in 10% neutral formalin solution and embedded in paraffin, and then the serial tissue sections were made 5㎛ in thickness and processed for immunohistochemical observation. The specimens were incubated with primary antibody against integrin α3 or integrin β1, each was diluted at 1 : 100, followed by the Supersensitive non-biotin horse radish peroxidase detection system with DAB as chromogen. After counterstaining with Gill's hematoxylin stain method and mounted, and examined under the light microscope. Based on the intensity of the immunoreactivity, intensity of the immunity was scored no epithelial stain, weak or focal epithelial stain, moderate or focal intensive epithelial stain, intense generalized epithelial staining for the epithelial, and connective tissue component in ameloblastomas, and normal oral mucosa on each. Attained results as follows. Expression of integrin α3 in the oral mucosa, weak reaction was noted on the all layers of epithelium, and submucosa. Expression of integrin β1 in the oral mucosa, intense reaction on the superficial layer, moderate reaction in basal layer were shown. Expression of integrin α3 in ameloblastomas, it was noted that weak reaction on the ameloblast like cells in the all types and rarely in basement membrane. Expression of integrin β1 in ameloblastomas, intense reaction on the tumor cell ,and partly in the nuclei in follicular type was noted, And moderate reaction on the tumor cell in plexiform , acathomatous types, but weak reaction in granular cell type was shown. This results result suggest that integrin α3 may influenced negligibly, but the integrin β1 influenced significantly the development of the ameloblastomas considering the response is increased on the region with highly cellular activities