The purpose 01' pl'esent study was to examine the molecular events in apoptosis by CoCl2, mimicking hypoxic cond ition and recovering effects by LED ir l'adiation on Human SH-SY5Y neuroblastoma cells The SOUl'ce 0 1' light for ir l'adiation was a continuous-wave LED emitting at a wavelenl양h of 590 nm, and manufactured that ene rgy density was 5 mW!cm2 on sample surface, After ir l'adiation, cell viabi lity was measured with BrdU , cell morphol ogy was examined with Diff- Quik staining, cell signaling was monitored with various apoptosis-related molecules using RNase Pl'otection Assay(RPA) , W11en treated with CoC12, apoptotic induction was found in the SH-SY5Y cells in a concentration-dependent and time-dependent manner , Diff-Quik s taining was revealed that DNA fragmentation re presented apoptosis was examined in CoC12-tl'eated group, Moreover, RPA assay of SH-SY5Y cclls lIs ing val'iolls apoptosis-related molecllles showed that the apoptotic cell population was mcreased J-loweve. there was sorne signifïcant change in LED irradiatied cells aftel' treatement of CoC12 The main mechanism for Lhese a poptosis appearecl to be mito c hondriεt - m ecliated pathway, such as cytochrome- c‘ caspase-9, caspase-3, pro-apototic protein ßax, anti-apototic protein Bcl-2, and death receptor• mediated pathway, such as Fas, cas pase- 8, a ncl TNFRl These results demonstrate that CoCI2 induce apoptosis in SH-SY5Y via different dual apop tosis pathway through death receptor pathway as well as mitochondria- dependent pathway and LED irradiation can recl llces the CoCl2-induced apoptosis by blocking their internal signaling pathway
The number 01' patien ts with tongue carcinoma is increasing rapidly among young indiv idua ls in many parts of the worl d. Until now‘ most of studies were focused on the comparison malignancy wi th normal 01' dysplasia. There is little report of gene a lterations in normal to cancer , oral carcinogenesis. The purpose of present study is to evaluate the gene a ltc ration in every steps of oral carcinogenes is by DD- PCR. To induce tongue carcinoma in rat by 4- NQO. each dri nking water made 10ppm. 25ppm. 50ppm and control(o nly D.W without 4-NQO) . Specimens were classified into 4 groups s uch as co ntrol, I(mild & mocle rate dysplasia) , II(severe dysplasia and car cinoma in s itu) , III(carcinoma). Total RNA was ext racted and DD- PCR was performed using customized random primers. And to confïrmed t he results of DD-PCR‘ RT- PCR a nd real-time PCR with specific primers were carried out. There was phenotypic alteration in tongue 。f dosc a nd t imc dc pcndcnt man ncr. In gross examination, multiple papules, patch form or ulcerations were observed during 4 - NQO t reatment Hi s tologicall y, dysplasia was observed in 3 to 6 month and tumor formation in 6 to 8 month For DD-PCR, RT-PCR and real-time PCR, cyclophilin A, BAC RP23-372MB and BAC CH230-103E9 were differ entia lly expressed. Taken together, cyclophilin A has a role in all steps of oral carcinogenesis. BAC RP23-372N田is implica ted in carcinoma in s itu a nd BAC CH230-103E9 mRNA expression is assoicated with dysplasia and carcinom in s itu Conclus ively. some genes a re impli catcd a ll st eps of oral carci nogenesis, others are associated with one step, whi ch meant that genes are di fferentia lly expressed in every steps
LEDs have been shown to be a safe, efficient, light-weight, and less-expensive alternative to heal wound. LED irradiation at the same biostimulatory wavelength of previous laser studies have similar biochemical effects The purpose of present study is to evaluate the effects of wound healing by LED irradiation. Thirty 34-day-old sprague dawley were used for present study. 1.5mm diameter defected holes were formed in both ear lobes of rat by rubber dam punch. 635nm and 890nm irradiation was performed by LED for 2 weeks, followed by histologíc examination staíned with H&E and Masson trichrome. Also, RT-PCR was carried out to find out the mRNA expression level in gingival fjbroblast irradiated by 635nm for 1 hour. In gross exarnination, wound healing was observed in irradiated group comparing to control For microscopic exarnination, repair by connective tissues was filled in defects of irradiated group, while dense cellular bands consisting of fjbroblasts and capi llaries were found at the end of defect in control By staining of masson trichrome, amount of collagens were found in irradiated group. In a result of RT-PCR, mRNA expressions of TGF- ß , MMP-1,3 and Timp-3 were down-regulated in irradiated group comparing with their expression in control group. Taken together, LED irradiation increase the prolifeation and the activity of fibroblasts and down-regualted the TGF-ß , MMP- 1,3 and Timp- 3 mRNA, followed by activation of would healing.