Purpose: To investigate the effect of deacetylase inhibitory trichostatin A (TSA) on anti HepG2 liver carcinoma cells and explore the underlying mechanisms. Materials and Methods: HepG2 cells exposed to different concentrations of TSA for 24, 48, or 72h were examined for cell growth inhibition using CCK8, changes in cell cycle distribution with flow cytometry, cell apoptosis with annexin V-FTIC/PI double staining, and cell morphology changes under an inverted microscope. Expression of ${\beta}$-catenin, HDAC1, HDAC3, H3K9, CyclinD1 and Bax proteins was tested by Western blotting. Gene expression for ${\beta}$-catenin, HDAC1and HDAC3 was tested by q-PCR. ${\beta}$-catenin and H3K9 proteins were also tested by immunofluorescence. Activity of Renilla luciferase (pTCF/LEF-luc) was assessed using the Luciferase Reporter Assay system reagent. The activity of total HDACs was detected with a HDACs colorimetric kit. Results: Exposure to TSA caused significant dose-and time-dependent inhibition of HepG2 cell proliferation (p<0.05) and resulted in increased cell percentages in G0/G1 and G2/M phases and decrease in the S phase. The apoptotic index in the control group was $6.22{\pm}0.25%$, which increased to $7.17{\pm}0.20%$ and $18.1{\pm}0.42%$ in the treatment group. Exposure to 250 and 500nmol/L TSA also caused cell morphology changes with numerous floating cells. Expression of ${\beta}$-catenin, H3K9and Bax proteins was significantly increased, expression levels of CyclinD1, HDAC1, HDAC3 were decreased. Expression of ${\beta}$-catenin at the genetic level was significantly increased, with no significant difference in HDAC1and HDAC3 genes. In the cytoplasm, expression of ${\beta}$-catenin fluorescence protein was not obvious changed and in the nucleus, small amounts of green fluorescence were observed. H3K9 fluorescence protein were increased. Expression levels of the transcription factor TCF werealso increased in HepG2 cells following induction by TSA, whikle the activity of total HDACs was decreased. Conclusions: TSA inhibits HDAC activity, promotes histone acetylation, and activates Wnt/${\beta}$-catenin signaling to inhibit proliferation of HepG2 cell, arrest cell cycling and induce apoptosis.
Lee Suk;Seong Jinsil;Kim Yong Bae;Cho Kwang Hwan;Kim Joo Ho;Jang Sae Kyung;Kwon Soo Il;Chu Sung Sil;Suh Chang Ok
Radiation Oncology Journal
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v.19
no.4
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pp.319-326
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2001
Purpose : Planning target volume (PTV) for tumors in abdomen or thorax includes enough margin for breathing-related movement of tumor volumes during treatment. Depending on the location of the tumor, the magnitude of PTV margin extends from 10 mm to 30 mm, which increases substantial volume of the irradiated normal tissue hence, resulting in increase of normal tissue complication probability (NTCP). We developed a simple and handy method which can reduce PTV margins in patients with liver tumors, respiratory motion reduction device (RRD). Materials and methods : For 10 liver cancer patients, the data of internal organ motion were obtained by examining the diaphragm motion under fluoroscope. It was tested for both supine and prone position. A RRD was made using MeV-Green and Styrofoam panels and then applied to the patients. By analyzing the diaphragm movement from patients with RRD, the magnitude of PTV margin was determined and dose volume histogram (DVH) was computed using AcQ-Plan, a treatment planning software. Dose to normal tissue between patients with RRD and without RRD was analyzed by comparing the fraction of the normal liver receiving to $50\%$ of the isocenter dose. DVH and NTCP for normal liver and adjacent organs were also evaluated. Results : When patients breathed freely, average movement of diaphragm was $12{\pm}1.9\;mm$ in prone position in contrast to $16{\pm}1.9\;mm$ in supine position. In prone position, difference in diaphragm movement with and without RRD was $3{\pm}0.9\;mm$ and 12 mm, respectively, showing that PTV margins could be reduced to as much as 9 mm. With RRD, volume of the irradiated normal liver reduced up to $22.7\%$ in DVH analysis. Conclusion : Internal organ motion due to breathing can be reduced using RRD, which is simple and easy to use in clinical setting. It can reduce the organ motion-related PTV margin, thereby decrease volume of the irradiated normal tissue.
Background: Because of the common etiologic factor, such as smoking, lung cancer and chronic obstructive pulmonary disease are often present in the same patient. The preoperative prediction of remaining pulmonary function after the resectional surgery is very important to prevent serious complication and postoperative respiratory failure. $^{99m}Tc$-MAA perfusion scan has been used for the prediction of postoperative pulmonary function, but it may be inaccurate in case of large V/Q mismatching. We compared $^{99m}Tc$-DTPA radioaerosol inhalation scan with $^{99m}Tc$-MAA perfusion scan in predicting postoperative lung function. Method: Preoperative inhalation scan and/or perfusion scan were performed and pulmonary function test were performed preoperatively and 2 month after operation. We predicted the postoperative pulmonary functions using the following equations. Postpneurnonectomy $FEV_1$=Preop $FEV_1x%$ of total function of lung to remain Postlobectomy $FEV_1$=Preop $FEV_1{\times}$(% of total 1-function of affected lung${\times}$$\frac{Number\;of\;segments\;to\;be\;resected}{Number\;of\;segments\;of\;affected\;lung})$ Results: 1) The inhalation scan showed good correlations between measured and predicted $FEV_1$, FVC and $FEF_{25-75%}$. (correlation coefficiency; 0.94, 0.91, 0.87 respectively). 2) The perfusion scan also showed good correlations between measured and predicted $FEV_1$, FVC and $FEF_{25-75%}$. (correlation coefficiency; 0.86, 0.72, 0.87 respectively). 3) Among three parameters, $FEV_1$ showed the best correlations in the prediction by lung scans. 4) Comparison between inhalation scan and perfusion scan in predicting pulmonary function did not show any significant differneces except FVC. Conclusion: The inhalation scan and perfusion scan are very useful in the prediction of postoperative lung function and don't make a difference in the prediction of pulmonary function a1though the former showed a better correlation in FVC.
Background: Preoperative 5-fluorouracil (5-FU)-based chemoradiotherapy is a standard treatment for locally advanced colorectal cancer (CRC). However, CRC cells often develop chemoradiation resistance (CRR). Recent studies have shown that long non-coding RNA (lncRNA) plays critical roles in a myriad of biological processes and human diseases, as well as chemotherapy resistance. Since the roles of lncRNAs in 5-FU-based CRR in human CRC cells remain unknown, they were investigated in this study. Materials and Methods: A 5-FU-based concurrent CRR cell model was established using human CRC cell line HCT116. Microarray expression profiling of lncRNAs and mRNAs was undertaken in parental HCT116 and 5-FU-based CRR cell lines. Results: In total, 2,662 differentially expressed lncRNAs and 2,398 mRNAs were identified in 5-FU-based CRR HCT116 cells when compared with those in parental HCT116. Moreover, 6 lncRNAs and 6 mRNAs found to be differentially expressed were validated by quantitative real time PCR (qRT-PCR). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis for the differentially expressed mRNAs indicated involvement of many, such as Jak-STAT, PI3K-Akt and NF-kappa B signaling pathways. To better understand the molecular basis of 5-FU-based CRR in CRC cells, correlated expression networks were constructed based on 8 intergenic lncRNAs and their nearby coding genes. Conclusions: Changes in lncRNA expression are involved in 5-FU-based CRR in CRC cells. These findings may provide novel insight for the prognosis and prediction of response to therapy in CRC patients.
This study was conducted to reveal stand structure and competition status by site types of natural deciduous forest in Pyungchang, Kangwon-do. The study site was divided by nine different types based on elevation(higher than 1,000m, 700~1,000m. lower than 700m) and topography(ridge, slope, valley). The objective of this study is to provide a basic information necessary for the environment-friendly management methods of natural deciduous forest on the basis of the stand structure and competition status by site types. It is confirmed that the range from 700m to 1,000m in elevation was the most suitable for stand growth. Species diversity and richness index also tended to be increased as elevation decreased. The ridge was the best in terms of stand growth by topography and followed by slope and valley in order. In addition, as expected, species with high importance value showed relatively low evenness index. The distance-independent competition index was selected as the best competition index model in seven site types of natural deciduous forest. On the other hand, the distance-dependent competition index was highly correlated with periodic annual increment of diameter in both ridge at higher than 1,000m and valley of 700m to 1,000m in elevation. It is proved, as a result, that the best competition index model is somewhat different by site types. From the analysis growth characteristics and competition status by site types, it is identified that the species with high importance value performed well in both growth and competition. The growth of Q. mongolica was excellent in the areas of higher elevation than 700m. Although K. pilus had relatively low importance value in higher elevation than 1,000m and ridge of 700m to 1,000m, the species had stronger competition status rather than other species. Also, U. davidiana and M. amurensis were good in competition status at lower elevation than 1,000m. It is necessary, therefore, that appropriate tending practice should be adopted based on the growth pattern and competition status of each species distributed by site types.
Tc-99m MIBI, a lipophilic cation, was reported as a useful agent for localization of lung cancer. The effect of radiation therapy on the uptake of Tc-99m MIBI in lung cancer, however, was not well evaluated. The aim of the present study was to elucidate the usefulness of Tc-99m MIBI SPECT in the localization and the assessment of radiotherapy in non-small cell lung cancer. Twenty patients(19 males and 1 female, mean age 59, 16 squamous cell ca and 4 adenoca) were studied with Tc-99m MIBI SPECT before radiation therapy. Eleven patients(10 males and 1 female, mean age 59, 8 squamous cell ca and 3 adenoca) were repeated the study 1 month after the completion of radiation therapy(mean dose 6453cGy). All patients showed positive uptakes of Tc-99m MIBI in their tumors. One patient showed a hot uptake in atelectatic area. There was no difference of Tc-99m MIBI uptakes between squamous cell ca and adenoca either on planar or tomographic images. Tc-99m MIBI uptake ratios of squamous cell ca and adenoca were $1.50{\pm}0.16$ and $1.45{\pm}0.15$ on planar images, and $2.73{\pm}0.46$ and $2.54{\pm}0.37$ on tomographic images, respectively. The concordance between radiological change(chest x-ray and CT) and change of Tc-99m MIBI uptakes was 9/11 (81.8% ). In conclusion, Tc-99m MIBI SPECT was useful in the localization of tumor and the assessment of radiation therapy in non-small cell lung cancer.
Kim, Myung-Hyun;Bang, Hea-Son;Han, Min-Su;Hong, Hey-Kyoung;Na, Young-Eun;Kang, Kee-Kyung;Lee, Jeong-Taek;Lee, Deog-Bae
Korean Journal of Environmental Agriculture
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v.28
no.2
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pp.125-130
/
2009
The aim of this study was to investigate whether differences in the distribution of soil invertebrates among different vegetation types (forest, reservoir, and crop land types) in rural area. A total of 18 orders and 137 species were collected by pitfall traps. Species numbers were the lowest (33 species) at the Chamaecyparis obtusa plantation (St. 6). On the forest sites, the individual number of Hymenoptera was the most abundant, and Acari and Coleoptera was the relatively more abundant than the other sites. On the reservoir sites (Salix chaenomeloides community), the individual number of Collembola was the most abundant, and Diptera was the relatively more abundant than the other sites. On the crop land sites, the individual numbers of Collembola, Hymenoptera, and Araneae were the relatively more abundant than the other orders. The density of Araneae was higher in the reservoir and crop land sites than in the forest sites. From a point of view of biodiversity, although the diversity index(H') was the highest in the mixed broad-leaved forest type (St. 2) with Quercus serrata and Q. acutissima, and the lowest in the upland levee of crop land(St. 11), there was no significant difference among the habitat or vegetation types. According to the community analysis, the soil invertebrates could be divided into 4 groups, the mixed broad-leaved forest type (A group), the plantation or pure forest type (B group), the reservoir type (C group), and the crop land type (D group).
Proceedings of the Korean Society for Food Science of Animal Resources Conference
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2004.05a
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pp.89-119
/
2004
This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.
This study was designed to identify the relationship between emotional labor and reaction factor(job satisfaction, fatigue), buffer factor(social support) among dental hygienist. The number of respondents was 50 who work in dental clinic and period of investigation was march 2010 through April 2010. A structured questionnaire was employed to evaluate the level of participants' sociodemographics, job-related factors, health-related behaviors, emotional labor and self-perceived fatigue, job satisfaction, social support. While the mean values of emotional labor were higher than thosewho work in hospital and lower than those who work in service employees. These results suggest that emotional labor was related to an increase in fatigue and strong correlation between involuntary emotional expression, self-perceived fatigue, job satisfaction, social support in dental hygienist. Multiple regression analysis has found that social support and no. of personnel were main factors that influence to the level of emotional labor in dental hygienist. The results of this study suggest that emotional labor was a determinant predictor of self perceived fatigue. Thus, a management program for emotional labor is strongly recommended for promotion of quality of life in dental hygienist and to improve dental service and competitiveness.
Cervical cancer is the fourth most frequently diagnosed cancer in women worldwide. In lower Human Development Index countries, it has the second highest incidence and mortality among cancer in women. Therefore, better diagnosis and treatment systems are needed. Among them, estrogen receptor alpha ($ER-{\alpha}$) mRNA expression has been analyzed with RT-qPCR since several studies reported that $ER-{\alpha}$ is necessary in the maturation of the uterus and is related to cervical cancer. In this study, $ER-{\alpha}$ quantitative analysis was performed on various lesions and normal tissue samples. Based on the receiver operating characteristic (ROC) curve, its sensitivity and specificity were 85% and 75%, respectively, showing higher or similar results to those of conventional HPV tests. In addition, its expression level was analyzed with clinical information. With regression analysis, the R square value between the $ER-{\alpha}$ mRNA expression level and menopause status was 0.5041, indicating a strong correlation. This study was performed as part of a pilot study and suggests that $ER-{\alpha}$ is related to carcinogenesis. Future studies will examine other hormones and menopausal factors with a larger sample size.
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