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.

Gemcitabine (Gemzar, 2,2-difluorodeoxycytidine, or dFdC) is an analog of cytosine arabinoside with anti-tumor activity in a few human cancers (lung, ovary, pancreatic and breast cancers). However, the mechanism of apoptosis by this compound in carcinoma has not been fully elucidated. In the present study, we investigated that gemcitabine alone and combination with cisplatin or 5-FU are cancer toxicity using lung cancer cell line A549 by MTT, FACS analysis, and Western blot assay. Also, to confirm enhanced antitumor activity in vivo using an xenograft tumor model. The MTT assay showed higher cytotoxic effect in combination with gemcitabine-cisplatin or gemcitabine-5-FU than gemcitabine alone. FACS analysis showed that gemcitabine-cisplatin combination increased hypodiploid DNA to 70.84 %. The induction of apoptosis showed more increase in combination with gemcitabine-cisplatin or gemcitabine-5-FU than gemcitabine alone. The Western results showed higher expression of p53 and p21WAF/CIP1 protein in combination treatment with gemcitabine-cisplatin or gemcitabine-5-FU than gemcitabine treatment alone. But, Bcl-2 protein expression decreased. In vivo experiments showed that more decreased tumor size and more increased survival rate on combination with gemcitabine- cisplatin or gemcitabine-5-FU combination than gemcitabine alone. In conclusion, this study suggests that gemcitabine combined with cisplatin or 5-FU are the synergistic effect of anticancer therapy on lung cancer.

The purpose of this study was to improve the professional's development of skill in qualitative analysis of humanMovement fromthe various subdisciplines of physical education. The conclusions were as follows; First, Physical educators and coaches can impr

This study was to investigate the influence of cooling on muscle force and viscoelastic properties of tendon structures in themedial gastrocnemius (MG) muscle. The subject was instructed to gradually increase force (10% MVC step) from a relaxed state to MVC within 3 s. At this time, it was measured by an ultrasonographic probe was attached and that an electrode was attached to monitor EMG. The F values at 50 100% of MVC were significantly greater under the cold condition than under the non-cold condition (p<.05). The values at 80~100% of MVC were significantly higher under the cold condition than under the non-cold condition (p<.05). The elongation under the non-cold condition had a tendency to be greater than that under the cold condition. The results suggest that cooling results in an increase in the stiffness of tendon structures with a reduction of muscle force and elongation.

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.