• Asian Pacific Journal of Cancer Prevention
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• v.15 no.18
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• pp.7575-7581
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• 2014

MicroRNAs (miRNAs) play an essential role in the development and progression of nasopharyngeal carcinomas (NPC). Despite advances in the field of cancer molecular biology and biomarker discovery, the development of clinically validated biomarkers for primary NPC has remained elusive. In this study, we investigated the expression and clinical significance of miRNAs as novel primary NPC diagnostic biomarkers. We used an array containing 2, 500 miRNAs to identify 22 significant miRNAs, and these candidate miRNAs were validated using 67 fresh NPC and 25 normal control tissues via quantitative real-time PCR (qRT-PCR). Expression and correlation analyses were performed with various statistical approaches, in addition to logistic regression and receiver operating characteristic curve analyses to evaluate diagnostic efficacy. qRT-PCR revealed five differentially expressed miRNAs (miR-93-5p, miR-135b-5p, miR-205-5p and miR-183-5p) in NPC tissue samples relative to control samples (p<0.05), with miR-135b-5p and miR-205-5p being of significant diagnostic value (p<0.01). Moreover, comparison of NPC patient clinicopathologic data revealed a negative correlation between miR-93-5p and miR-183-5p expression levels and lymph node status (p<0.05). These findings display an altered expression of many miRNAs in NPC tissues, thus providing information pertinent to pathophysiological and diagnostic research. Ultimately, miR-135b-5p and miR-205-5p may be implicated as novel NPC candidate biomarkers, while miR-93-5p, miR-650 and miR-183-5p may find application as relevant clinical pathology and diagnostic candidate biomarkers.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.11
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• pp.6341-6345
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• 2013

To clarify the role of stem cells in hepatocarcinogenesis, glypican-3 (GPC-3) and E-cadherin expression was investigated in embryonic cell lineages. Mouse embryonic stem cells (ESCs), hepatic progenitor cells (HPCs) and hepatocyte like cells (HCs), representing 0, 22 and 40 days of differentiation, respectively, were treated in vitro with diethylnitrosamine (DEN) at four doses (0, 1, 5 and 15 mM; G1, G2, G3 and G4, respectively) for 24 h and GPC-3 and E-cadherin expression was examined by relative quantitative real-time PCR and immunocytochemistry. GPC-3 mRNA expression was significantly different for G4 at day 0 (p<0.001) and for G4 at day 22 (p<0.01) compared with the control (G1). E-cadherin mRNA expression was significantly different for G3 and G4 at day 0 (p<0.05 and p<0.001, respectively), for G2 and G4 (p<0.05 and p<0.001, respectively) at day 22 and for G2 and G4 (p<0.01 and p<0.001, respectively) at day 40 compared with G1. Immunofluorescence staining for GPC-3 showed a membranous and/or granular expression in cytoplasm of ESCs and HPCs and granular and/or diffuse expression in cytoplasm of HCs, which were also stained by E-cadherin. DEN treatment increased GPC-3 expression in ESCs, HPCs and HCs, with increase of E-cadherin expression. Taken together, the expression of GPC-3 was altered by DEN treatment. However, its expression pattern was different at the stage of embryo stem cells and its derived hepatic lineage cells. This suggests that GPC-3 expression may be modulated in the progeny of stem cells during their differentiation toward hepatocytes, associated with E-cadherin expression.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.409-417
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• 2012

We identified two novel expansin (EXP) genes in the expressed sequence tag database of Bursaphelenchus xylophilus, designated as Bx-EXPB2 and -EXPB3. Novel Bx-EXPBs encoded 150 amino acids and their similarities in coding sequence were 70.7-84.0% to the previously reported EXPB1 of B. xylophilus. Bx-EXPB2 and Bx-EXPB3 were clustered with Bx-EXPB1 and Bm-EXPB1, respectively, forming the independent phylogeny with other nematode EXPs. All identified Bx-EXPBs contained the signal peptide and were only expressed during the propagative stage, suggesting that they are secreted to facilitate nematode migration through hosts by loosening cell walls during infection. Quantitative real-time PCR analysis showed that the relative accumulation of Bx-EXPB3 mRNAs was the highest among the three Bx-EXPs examined and the order of mRNA accumulation was as follows: Bx-EXPB3 > Bx-EXPB2 >> Bx-EXPB1. Homology modeling of Bx-EXPBs showed that the structurally optimum template was EXLX1 protein of Bacillus subtilis, whichshared residues essential for catalytic activity with Bx-EXPB1 and Bx-EXPB2 except for Bx-EXPB3. Taken together, Bx-EXPB1 and Bx-EXPB2 may be involved migration through plant tissues and play a role in pathogenesis.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.10
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• pp.5915-5920
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• 2013

Aim: Although aberrant miRNA expression has been documented, altered miR-101 expression in cervical cancer and its carcinogenic effects and mechanisms remain unexplored. The aim of our study was to investigate the role of miR-101 alteration in cervical carcinogenesis. Methods: Expression of miR-101 was examined by quantitative real-time reverse transcriptase PCR (qRT-PCR) in Hela cells. After modulating miR-101 expression using miR-101 mimics, cell growth, apoptosis and proliferation, and migration were tested separately by MTT or flow cytometry and cell wound healing assay and protein expression was detected by qRT-PCR. The expression of COX-2 in Hela cell was also examined by immunohistochemical staining and the correlation with miR-101 expression was analysed. Results: The miR-101 demonstrated significantly low expression in Hela cell. When we transfected miR-101 mimics into Hela cells, the modulation of miR-101 expression remarkably influenced cell proliferation, cycling and apoptosis: 1) The expression of microRNA-101 tended to increase after transfection; 2) Overexpression of miR-101 was able to promote cell apoptosis, the apoptosis rate being markedly higher (97.6%) than that seen pre-transfection (12.2%) (P<0.05); 3) The miR-101 negatively regulates cell migration and invasion, scratch results being lower ($42.7um{\pm}2um$) than that observed pre-transfection ($181.4um{\pm}2um$); 4) miRNA-101 inhibits the proliferation of Hela cells as well as the level of COX-2 protein, which was negatively correlated with miR-101 expression. Conclusions: Overexpression of miR-101 has obvious inhibitory effects on cell proliferation, migration and invasion. Thus reduced miR-101 expression could participate in the development of cervical cancer at least partly through loss of inhibition of target gene COX-2, which probably occurs in a relative late phase of carcinogenesis. Our data suggest an important role of miR-101 in the molecular etiology of cancer and indicate potential application of miR-101 in cancer therapy.

• Journal of Gastric Cancer
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• v.16 no.4
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• pp.221-229
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• 2016

Purpose: Dysregulated microRNAs (miRNAs) can contribute to cancer development by leading to abnormal proliferation of cells, apoptosis, and differentiation. Although several miRNAs that are related to gastric cancer have been identified, the reported results have been inconsistent. The aim of this study was to determine miRNA expression profiles and validate miRNAs up- and down-regulated in gastric cancer. Materials and Methods: We evaluated 34 primary gastric cancer tissues and paired adjacent nontumorous gastric tissues. Total RNA was extracted, and low-molecular-weight RNAs (<200 nucleotides) were isolated for further analysis. Two pairs of tissues were processed for GeneChip microarray analysis, and the identified up- and down-regulated miRNAs were validated by real-time quantitative polymerase chain reaction (qPCR). Results: In the set of differentially expressed miRNAs, 5 were overexpressed by more than 2 fold, and 5 were reduced by 2 fold or less in gastric cancer tissues compared with normal gastric tissues. Four of these miRNAs (miR-196b-5p, miR-375, miR-483-5p, and miR-486-5p) were then validated by qPCR, and the relative expression levels of 2 miRNAs (miR-196b-5p and miR-375) were significantly different between cancer and normal tissues. Conclusions: Our results revealed that the expression of miR-196b-5p and miR-375 significantly correlates with gastric cancer. These miRNAs could therefore serve as diagnostic biomarkers of gastric cancer.

• Tuberculosis and Respiratory Diseases
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• v.60 no.2
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• pp.151-159
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• 2006

Background : Excision repair cross complementing gene 1 (ERCC1) not only has a protective role against carcinogens, but plays an important role in cisplatin-resistance via the repair of cisplatin-DNA adducts. This study investigated the association between the ERCC1 expression levels in sputum and survival after cisplatin-based chemotherapy in patients with inoperable non-small cell lung cancer (NSCLC). Methods : Using the sputum collected from 67 inoperable (stage IIIa-IV) NSCLC patients treated with either taxanes (33 cases) or gemcitabine (34 cases) plus cisplatin, the relative expression levels of ERCC1 and the expression of the tumor specific antigen, MAGE, were examined by the quantitative RT-PCR and RT-PCR, respectively. The response and survival were compared with the relative level of ERCC1 or MAGE expression and the treatment modality. Results : In the sputum, ERCC1 and MAGE was detected in 74.6% and 40.2% of patients, respectively. Using the median ERCC1 level, the patients were classified as having high or low ERCC1 expression. The median overall survival (MST) was significantly longer in patients with a high ERCC1 expression level than those with a low expression level (84 weeks vs. 44 weeks respectively, P=0.017). In the taxene-based treatment group, the MST was longer than the gemcitabine group (79 weeks vs. 47 weeks, respectively, P=0.03). The levels of ERCC1 were significantly higher in patients who were MAGE-positive (P=0.003). In the MAGE-negative patients, the MST was longer in the high ERCC1 group (103 weeks vs. 43 weeks, P=0.008), but not in the MAGE-positive patients (62 weeks vs. 44 weeks, P=0.348). Conclusion : ERCC1 expression in the sputum can be a prognostic factor for survival after chemotherapy in patients with inoperable NSCLC.

• Food Science and Preservation
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• v.20 no.5
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• pp.699-704
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• 2013

Buckwheat Soksungjang (BS) is a bealmijang manufactured with buckwheat and soybeans. We manufactured BS using Bacillus subtilis HJ18-4 (HJ18-4), which has high enzyme activities and antibacterial effects. HJ18-4 was inoculated in a different process during the BS manufacturing, which was the meju-making time (Treat 1), and the salt water time was added (Treat 2). The physiochemical and microbial characteristics of the BS were analyzed. As a result, the total aerobic counts (7~8 log CFU/mL) in the BS increased after 15 days of fermentation. Especially, Treat 1 showed higher total aerobic counts and amino-type nitrogen (65.38~202.52 mg%) than Treat 2. During the BS fermentation, the reduction of the sugar contents and the enzyme (protease and amylase) activities decreased. In the relative quantitative expression level of PlcR, Treat 1 did not show toxin gene expressions at the end of the fermentation on Day 23. Treat 1 showed suitable B. cereus physiochemical quality characteristics and inhibition effects. When the modified-form type of fermented soybean paste was manufactured with a single starter, it could not reproduce the natural fermentation quality. These results suggest that the addition of a starter (HJ18-4) in the Meju manufacturing process could enhance the quality characteristics of the manufactured BS via natural fermentation and by suppressing B. cereus.

• Reproductive and Developmental Biology
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• v.30 no.2
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• pp.125-133
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• 2006

This study was conducted to detect the apoptosis incidence in blastocysts and to compare the abundance of Bax, Bcl2L1, VEGF and FGFR2 in in vitro fertilized (IVF), parthenogenetic (PAT) and nuclear transfer (NT) embryos. Oocytes matured for 40 hr were enucleated and reconstructed with confluenced fetal fibroblasts (FFs) derived from a ${\sim}45$ day fetus. Reconstructed eggs were then fused with 2 DC pulses (2.0 kV/cm, $30{\mu}sec$) and cultured with $7.5{\mu}g/ml$ cytochalasin B for 3 hr. Parthenotes (PAT) were produced with the same electric strength and culture for NT eggs. The embryos were cultured in NCSU-23 medium at $39^{\circ}C,\;5%\;CO_2,\;5%\l;O_2$ in air. In 3 runs, set of 10 embryos at the 4-cell to blastocyst stages were used to extract total RNA for analyzing the gene expression patterns of pro-apoptotic (Bax), anti-apoptotic (Bcl2L1), vasculogenesis (VEGF), implantation (FGFR2III) using real-time quantitative PCR. Cleavage and blastocyst rates were significantly higher (P<0.05) in IVF and PAT ($79.3{\pm}8.5\;and\;25.5{\pm}6.1,\;and\;85.0{\pm}6.4\;and\;38.6{\pm}5.5$, respectively)than NT counterparts ($65.1{\pm}5.2\;and\;15.6{\pm}3.0$, respectively). Significantly higher (P<0.05) total cells were observed in IVF controls and PAT ($34.7{\pm}5.8\;and\;38.1{\pm}4.1$) than NT embryos ($24.8{\pm}3.2$). Apoptosis index was significantly lower (P<0.05) in IVF than NT embryos. The Relative abundances (RA) of Bax and VEGF were significantly higher (P<0.05) at blastocyst stage in NT than IVF control. The RA of Bcl2L1 and FGFR2III were significantly higher (P<0.05) at blastocyst stage in IVF than NT. The present study observed the abnormal gene expressions in NT embryos at various developmental stages, suggesting certain clues to find out the cause of the low efficiency of NT to term.

• Journal of Veterinary Clinics
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• v.33 no.4
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• pp.194-199
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• 2016

The objective of this study was to examine the effect of trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) on the expression of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) pathway in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs). t10c12-CLA was treated with different concentrations in culture medium of LPS$na{\ddot{i}}ve$ and LPS-stimulated PBMCs. The mRNA expressions of prostaglandin $E_2$ ($PGE_2$)-synthase, COX-2 and 5-LOX were measured using quantitative real-time PCR. In addition, the production levels of $PGE_2$ and 5-LOX in culture supernatant from PBMCs with or without LPS were assessed by ELISA. In LPS$na{\ddot{i}}ve$ PBMCs, treatment of t10c12-CLA significantly (p < 0.05) increased the mRNA expressions of PGE2 synthase and 5-LOX compared to vehicle control. Expression of COX-2 mRNA did not show significant difference compared to vehicle control by t10c12-CLA treatment in LPS$na{\ddot{i}}ve$ PBMCs. However, the addition of LPS in PBMCs markedly (p < 0.05) increased the mRNA expression of COX-2, $PGE_2$ synthase and 5-LOX, and also significantly (p < 0.05) enhanced the production of $PGE_2$ and 5-LOX relative to LPS$na{\ddot{i}}ve$ PBMCs, respectively. However, the addition of t10c12-CLA significantly (p < 0.01) suppressed the LPS-induced excessive expression of COX-2, $PGE_2$ synthase, and 5-LOX compared to those of PBMCs treated with LPS alone. The production levels of $PGE_2$ and 5-LOX in culture supernatant from LPS-stimulated PBMCs were also significantly (p < 0.05) inhibited by the treatment of t10c12-CLA compared to LPS alone. These results suggested that t10c12-CLA has an anti-inflammatory effect via dual inhibition of COX-2 and 5-LOX with gene expression and production level in LPS-stimulated porcine PBMCs. Therefore, it was thought that t10c12-CLA can attenuate the inflammatory response by down-regulation of eicosanoids production.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.9
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• pp.1378-1386
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• 2013

Atopic dermatitis (AD) is a common skin disease characterized by chronic and relapsing inflammatory dermatitis with immunological disturbances. In spite of the continuous increase in the incidence of AD, it is regrettable that till date there is no effective treatment to treat the same. Therefore, the present study was designed to examine the possible anti-atopic effects of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (FCS) in 2,4-dinitrochlorobenzene (DNCB) induced AD in NC/Nga mice. Based on the results of HPLC analysis, we found that FCS contains anti-inflammatory factors such as gallic acid (10.18 mg/g) and ellagic acid (2.14 mg/g). The groups that we have used in this study included 0.1%, 1%, 5% fermented Castanea crenata inner shell extracts (FCS 0.1, FCS 1, FCS 5), 1,3-butylene glycol treated control (AD), and normal mice. After topical FCS treatment, we observed that the clinical severity score for AD was lower in both the FCS 1 and FCS 5 groups than the AD group. We also proved beyond doubt that there was improvement of melanin, erythema and skin moisture indices in the FCS 5 group. Spleen index and gene expression levels of pro-inflammatory cytokines such as IL-$1{\beta}$ and TNF-${\alpha}$ were significantly decreased in the FCS 5 group compared to the AD group (P<0.05). Further, we also found that the level of serum immunoglobulin E (IgE) in the FCS-treated group was decreased in a concentration-dependent manner. The results of our study suggest that FCS can be effectively used as a cosmeceutical ingredient for both the prevention and improvement of AD.