• Journal of Oral Medicine and Pain
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• v.38 no.4
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• pp.291-298
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• 2013

On this study, we treated rats with restraint stress, and observed the changes with an optical microscope. Within the salivary gland tissue, we measured cell apoptosis cycle evaluation which show positive reaction on TUNEL assay, and compared within the groups. For this study, 18 rats were divided into 3 groups; 1) 2 rats of group I were selected as a normal control. 2) 2 rats of group II, as a experimental control were placed in the restraint cone for 2 hours 3) 14 rats of group III were placed in the restraint cone for 2 hours once a day. The rats were sacrificed immediately (group II, as a experimental control), 1, 2, 3, 4, 5, 6 and 7days after application of the stress and the both parotid glands were excised. The conclusions follow. 1. 5 days after giving an confining stress to the parotid gland of Rats, we can observe the hypotropy and pus and inflammation of Rat parotid gland acinar cells, and after 7 days, we can see a cell apoptosis. 2. Through the In situ DNA end labeling assay and TUNEL dye, on serous glands, benign tumor cell increased with statistically significant result after 5 days from confining stress. And the index shows maximum value on 7th days, which is same result with histological opinion. Therefore, our study shows that a cell apoptosis can be induced by restraint stress on salivary gland tissue, and we think more study should be accomplished about the cell signaling pathway in the future.

• Journal of Korean Neurosurgical Society
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• v.38 no.3
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• pp.215-220
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• 2005

Objective : Many researchers believe that the hypothermia shows neuro-protective effect on brain injury. To understand the molecular mechanism of the hypothermic treatment, this study investigated its effects on the expression of cell death or survival related proteins such as p53, Bcl-2 and Bax in the rat traumatic brain injury[TBI] model. Methods : Twenty rats [Spraque Dawley, $200{\sim}250g$] were subjected to the brain injury of moderate severity [$2.4{\sim}2.6atm$] using the fluid percussion injury device and five rats were received only same surgery as controls. During 30minutes after the brain injury, the hypothermia group was maintained the body temperature around $34^{\circ}C$ while the control group were maintained that of $36^{\circ}C$. Five rats in each group were sacrificed 12h or 24h after brain injury and their brain sections was analyzed for physical damages by H-E stains and the extent of apoptosis by TUNEL assay and immunohistochemical stains. The tissue damage after TBI was mainly observed in the ipsilateral cortex and partly in the hippocampus. Results : Apoptosis was observed by TUNEL assay and the Bax protein was detected in both sample which harvested 12h and 24h after TBI. In the hypothermia treatment group, tissue damage and apoptosis were reduced in HE stains and TUNEL assay. In hypothermia treatment group rat shows more expression of the Bcl-2 protein and shows less expression of the Bax protein, at both 12h and 24h after TBI. Conclusion : These results show that the hypothermia treatment is an effective treatment after TBI, by reducing the apoptotic process. Therefore, it could be suggested that hypothermia has a high therapeutic value for treating tissue damages after TBI.

• Journal of Life Science
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• v.21 no.10
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• pp.1494-1499
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• 2011

${\beta}$-lapachone, a quinone of lapachol extracted from the bark of the lapacho tree, has been found to induce apoptosis in various human cancer cells. In the present study, we investigated further possible mechanisms by which ${\beta}$-lapachone exerts its pro-apoptotic action in cultured human lung cancer A549 cells. ${\beta}$-lapachone treatment resulted in inhibition of growth and induction of apoptosis in a concentration-dependent manner, as determined by MTT assay and flow cytometry analysis. The induction of apoptosis by ${\beta}$-lapachone was associated with up-regulation of the expression of p53 and p21 in both transcriptional and translational levels, and the phosphorylation of p53. In addition, ${\beta}$-lapachone activated caspase-3 and -9, and induced degradation of caspase-3 target proteins such as poly (ADP-ribose) polymerase (PARP) and ${\beta}$-catenin. Furthermore, ${\beta}$-lapachone treatment caused a progressive decrease in the expression levels of cyclooxygenase (COX)-2 without significant changes in the levels of COX-1, which was correlated with a decrease in prostaglandin E2 synthesis. Taken together, these results indicated that ${\beta}$-lapachone may have therapeutic potential in human lung cancer treatment.

• Radiation Oncology Journal
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• v.26 no.3
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• pp.173-180
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• 2008

Purpose: The modification of radiation-induced apoptosis by EGCG, known as antioxidants or oxidants, was studied in mice spleens irradiated with a lethal dose. Materials and Methods: Male C57BL/6 mice were divided into control, irradiation-only, and EGCG (100 mg/kg i.p. 1 h before irradiation) pretreatment groups. The mice were irradiated with a single whole-body dose of 7 Gy. The apoptosis in the spleens after irradiation of the lethal dose were analyzed by TUNEL assay. In addition, the expression levels of the Bax and Bcl-2 proteins were quantified using a Western blotting method. Results: The induction of apoptosis was detected in the splenic white pulp. The highest level of apoptosis was detected at 8 hours after irradiation. No significant difference was identified by the apoptotic index (53.9% vs. 52.1%, p=0.328) and relative Bax protein expression (0.86 vs. 0.81, p=0.335), between the irradiation-only and EGCG pretreatment group, respectively. However, a lower Bax/Bcl-2 ratio (1.64 vs. 0.97, p=0.037) and higher relative expression level of Bcl-2 protein (0.57 vs. 0.82, p=0.037) was measured in the EGCG pretreatment group. Conclusion: The EGCG pretreatment neither decreased the radiation-induced apoptosis in mice splenocytes, nor induced additional apoptosis.

• Korean journal of applied entomology
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• v.55 no.3
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• pp.267-275
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• 2016

Chlorine dioxide has an insecticidal activity via its production of reactive oxygen species (ROS). Its cytotoxic activity has been regarded as a main cause of the insecticidal activity. This study tested a hypothesis that cytotoxicity of chlorine dioxide is resulted from its induction of apoptosis against target cells using ROS. Injection of chlorine dioxide significantly reduced total hemocyte counts of Plodia interpunctella larvae and subsequently killed the larvae. To analyze the cytotoxicity with respect to apoptosis, terminal deoxyribonucleotidyl transferase nick end translation (TUNEL) assay was performed. An insect cell line (Sf9) cells were exposed to different concentrations of chlorine dioxide. TUNEL assay showed that chlorine dioxide induced significant apoptosis of Sf9 cells in a dose-dependent manner. When different concentrations of chlorine dioxide were injected to larvae of P. interpunctella, it showed a dose-dependent induction of apoptosis against hemocytes. However, addition of vitamin E significantly suppressed the apoptosis induction and insecticidal activity of chlorine dioxide in a dose-dependent manner. These results suggest that cytotoxicity of chlorine dioxide is resulted from its induction of apoptosis against insect cells using ROS.

• Journal of Physiology & Pathology in Korean Medicine
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• v.24 no.4
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• pp.661-667
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• 2010

Astragali Radix (AR) has been used in traditional medicinal herb, which is known to tonify the qi and blood. It has been asserted to be a tonic that can improve the function of the lungs, adrenal glands and the gastrointestinal tract, increase metabolism, promote healing and reduce fatigue. Also, Plantago asiatica (PA) has the anti-inflammatory, antiseptic, antitussive, cardiac, diuretic, expectorant, haemostatic effects, though it is considered to be less powerful than the seed. Recent research is showing that it is promising in lowering cholesterol and controlling diabetes. Obesity is characterized at the cellular level by an increase in the number and size of adipocytes differentiated from fibroblastic preadipocytes in adipose tissue. In the present study, we investigated the effects of 3 to 7 extracts of mixed medicinal herbs (water and ethanol extract of Astragali Radix; ARW and ARE, water and ethanol extract of Plantago asitica; PAW and PAE, and those mixed extracts; ARW+PAE, ARW+PAW and ARE+PAE) on adipogenesis and apoptosis in 3T3-L1 adipocytes by MTT assay, Oil-Red-O staining and TUNEL assay. And then, we determined total contents of phenolic compounds and flavonoid compounds in mixed herbal extracts. These result showed that herbal extracts (ARW, PAE and ARW+PAE) affected on adipogenesis and apoptosis in 3T3-L1 adipocytes among 3 to 7 extracts tested. Especially herbal mixed extracts (ARW+PAE) had the highest inhibition on differentiation of preadipocytes and lipoid accumulation of maturing adipocytes. And it did not affect cell viability of mature adipocytes, but herbal mixed extracts (ARW+PAE) increased apoptosis in mature adipocytes, as proved by highest concentration $200\;{\mu}g/m{\ell}$ using TUNEL assay. In addition, contents of total phenoilc compounds and flavonoids showed the highest level in ARE+PAE and PAE than ARW. These results suggest that this main target for adipocyte functions could be effective in improving the metabolic syndrome including obesity.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.4
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• pp.306-311
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• 2005

Despite advances in surgery, radiotherapy, and chemotherapy, the survival of patients with oral squamous cell carcinoma has not significantly improved over the past several decades. Gene therapy is currently under investigation and shows us new possibility of cancer curing method. This experiment was undergone to find out the cell growth inhibition effect and evidence of apoptosis by HCCS-1(human cervical cancer suppressor-1), one of the candidates of tumor suppressor gene, transducted to human oral cancer cell line. To determine the efficiency of the adenovirus as a gene delivery vector cell line was transducted with LacZ gene and analysed with X-gal staining. Northern blot was performed to confirm the transfection with HSCC-1 gene and cell viability was assessed by cell cytotoxicity assay using cell count kit(CCK). To show the evidence of apoptosis, DNA fragmentation assay and flow cytometry(FACS) were performed. We had successfully construct the recombinant HSCC-1 adenovirus(Ad5CMV-HCCS-1), and importation efficiency was 20% at 2 MOI(multiplicity of infection), 80% at 20 MOI. Northern blot analysis showed that a single 0.6kb mRNA transcript was expressed in Ad5CMV-HCCS-1 transducted cell lines. As a result of CCK, when comparing to control subjects, transducted group showed 50% growth inhibition. In DNA fragmentation assay, according to increasing of MOI, DNA volume was diminished. In FACS analysis, DNA distribution showed fragmentation. This results imply that HCCS-1gene has growth inhibition effect in human oral cancer cell lines through apoptosis induction.

• Molecules and Cells
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• v.37 no.11
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• pp.785-794
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• 2014

Mitophagy, a cellular process that selectively targets dysfunctional mitochondria for degradation, is currently a hot topic in research into the pathogenesis and treatment of many human diseases. Considering that hypoxia causes mitochondrial dysfunction, which results in cell death, we speculated that selective activation of mitophagy might promote cell survival under hypoxic conditions. In the present study, we introduced the Regulator of calcineurin 1-1L (Rcan1-1L) to initiate the mitophagy pathway and aimed to evaluate the effect of Rcan1-1L-induced mitophagy on cell survival under hypoxic conditions. Recombinant adenovirus vectors carrying Rcan1-1L were transfected into human umbilical vein endothelial cells and human adult cardiac myocytes. Using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay and Trypan blue exclusion assay, Rcan1-1L overexpression was found to markedly reverse cell growth inhibition induced by hypoxia. Additionally, Rcan1-1L overexpression inhibited cell apoptosis under hypoxic conditions, as detected by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay. Meanwhile, the mitochondria-mediated cell apoptotic pathway was inhibited by Rcan1-1L. In contrast, knockdown of Rcan1-1L accelerated hypoxia-induced cell apoptosis. Moreover, Rcan1-1L overexpression significantly reduced mitochondrial mass, decreased depolarized mitochondria, and downregulated ATP and reactive oxygen species production. We further delineated that the loss of mitochondrial mass was due to the activation of mitophagy induced by Rcan1-1L. Rcan1-1L overexpression activated autophagy flux and promoted translocation of the specific mitophagy receptor Parkin into mitochondria from the cytosol, whereas inhibition of autophagy flux resulted in the accumulation of Parkin-loaded mitochondria. Finally, we demonstrated that mitochondrial 1permeability transition pore opening was significantly increased by Rcan1-1L overexpression, which suggested that Rcan1-1L might evoke mitophagy through regulating mitochondrial permeability transition pores. Taken together, we provide evidence that Rcan1-1L overexpression induces mitophagy, which in turn contributes to cell survival under hypoxic conditions, revealing for the first time that Rcan1-1L-induced mitophagy may be used for cardioprotection.

• Journal of Food Hygiene and Safety
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• v.27 no.4
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• pp.406-414
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• 2012

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-${\alpha}$ assays were performed in Raw 264.7 cell. Treatment with fucoidan significantly increased production of NO and TNF-${\alpha}$, 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-VAD-FMK, 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.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.2
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• pp.625-631
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• 2012

In order to investigate whether SKI-II could reverse drug resistance and its possible mechanisms, we treated SGC7901/DDP cells with SKI-II or SKI-II in combination with DDP. Then cell growth, apoptosis, micromorphological changes, and expression of SphK1, P-gp, NF-${\kappa}B$, Bcl-2 and Bax were assessed by MTT assay, flow cytometry, electron microscopy, immunocytochemistry and Western blot assay respectively. SGC7901/DDP cells were insensitive to cisplatin 2.5mg/L, but when pretreated with SKI-II, their proliferation was inhibited by cisplatin 2.5mg/L significantly, the inhibition rate increasing with time and dose. The apoptosis rate was also significantly elevated. Expression of SphK1 and P-gp was decreased significantly, Pearson correlation analysis showing significant correlation between the two (r=0.595, P<0.01). Expression of NF-${\kappa}B$ and Bcl-2 was decreased significantly,while that of Bax was increased, compared to the control group. There were significant correlations between SphK1 and NF-${\kappa}B$(r=0.723, P<0.01), NF-${\kappa}B$ and Bcl-2(r=0.768, P<0.01). All these data indicated that SKI-II could reverse drug resistance of SGC7901/DDP to cisplatin by down-regulating expression of P-gp and up-regulating apoptosis through down-regulation of SphK1. The increased apoptotic sensitivity of SGC7901/DDP to cisplatin was due to the decreasing proportion of Bcl-2/Bax via down-regulating NF-${\kappa}B$.