• 대한약침학회지
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• 제19권1호
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• pp.59-69
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• 2016

Objectives: The mushroom Ganoderma lucidum has been widely used as a traditional herbal medicine for many years. Although several studies have focused on the anti-oxidative activity of this mushroom, the molecular mechanisms underlying its activity have not yet been clearly established. The present study investigated the cytoprotective effect of ethanol extract of Ganoderma lucidum (EGL) against oxidative stress (hydrogen peroxide, $H_2O_2$) and elucidated the underlying mechanisms in a C2C12 myoblast cell line. Methods: Oxidative stress markers were determined by using the comet assay to measure reactive oxygen species (ROS) generation and deoxyribonucleic acid (DNA) damage. Cell viability and Western blotting analyses were employed to evaluate the cellular response to EGL and $H_2O_2$ in C2C12 cells. Transfection with nuclear factor erythroid 2-related factor 2 (Nrf2)-specific small interfering ribonucleic acid (siRNA) was conducted to understand the relationship between Nrf2 expression and $H_2O_2$-induced growth inhibition. Results: The results showed that EGL effectively inhibited $H_2O_2$-induced growth and the generation of ROS. EGL markedly suppressed $H_2O_2$-induced comet-like DNA formation and phosphorylation of histone H2AX at serine 139 ($p-{\gamma}H2AX$), a widely used marker of DNA damage, suggesting that EGL prevented $H_2O_2$-induced DNA damage. Furthermore, the EGL treatment effectively induced the expression of Nrf2, as well as heme oxygenase-1 (HO-1), with parallel phosphorylation and nuclear translocation of Nrf2 in the C2C12 myoblasts. However, zinc protoporphyrin IX, a HO-1 inhibitor, significantly abolished the protective effects of EGL against $H_2O_2$-induced accumulation of ROS and reduced cell growth. Notably, transient transfection with Nrf2-specific siRNA attenuated the cytoprotective effects and HO-1 induction by EGL, indicating that EGL induced the expression of HO-1 in an Nrf2-dependent manner. Conclusion: Collectively, these results demonstrate that EGL augments the cellular anti-oxidant defense capacity through activation of Nrf2/HO-1, thereby protecting C2C12 myoblasts from $H_2O_2$-induced oxidative cytotoxicity.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2987-2991
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• 2015

Background: The prognostic role of thyroid transcription factor-1 (TTF-1) expression in lung cancer has been assessed but with inconsistent results. The present study aimed to evaluate the prognostic value of TTF1 expression in advanced non-squamous non-small cell lung cancer (NSCLC). Materials and Methods: In this retrospective study, patients with stage IIIB-IV non-squamous NSCLC were enrolled. Progression free survival (PFS) and overall survival (OS) were assessed according to TTF1 expression status, age categories (${\leq}60$ vs >60 years), gender, performance status (PS) (0-2 vs 3-4), type of 1st line chemotherapy (pemetrexed containing vs others) and EGFR status. Results: A total of 120 patients were included. In univariate analysis, PFS was improved in patients with PS 0-2 (7.0 vs 2.0 months, p=0.002) and those who received pemetrexed-containing chemotherapy (9.2 vs 5.8 months, p=0.004). OS was improved in female patients (23.0 vs 8.7 months, p<0.0001), PS 0-2 (14.4 vs 2.0 months, p<0.0001), those with pemetrexed-containing chemotherapy (17.0 vs 11.0 months, p=0.019), TTF1-positive (12.8 vs 5.8 months, p=0.011) and EGFR- mutant patients (23.0 vs 11.7 months, p=0.006). In multivariate analysis, male gender (HR=2.34, p=0.025) and non-pemetrexed containing therapy (HR=2.24, p=0.022) were independent predictors of worse PFS. Wild EGFR status (HR=2.49, p=0.015) and male gender (HR=2.78, p=0.008) were predictors of worse OS. Conclusions: Pemetrexed-containing therapy significantly improved PFS while OS was improved in EGFR mutant patients. Female patients had better PFS and OS. TTF1 expression was not a prognostic marker in advanced non-squamous NSCLC.

• Asian Pacific Journal of Cancer Prevention
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• 제16권9호
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• pp.4065-4069
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• 2015

Background: Epidermal growth factor-like domain multiple 7 (EGFL7), a secreted protein specifically expressed by endothelial cells during embryogenesis, recently was identified as a critical gene in tumor metastasis. Epithelial-mesenchymal transition (EMT) was found to be closely related with tumor progression. Accordingly, it is important to investigate the migration and EMT change after knock-down of EGFL7 gene expression in human pancreatic cancer cells. Materials and Methods: EGFL7 expression was firstly testified in 4 pancreatic cancer cell lines by real-time polymerase chain reaction (Real-time PCR) and western blot, and the highest expression of EGFL7 was found in PANC-1 cell line. Then, PANC-1 cells transfected with small interference RNA (siRNA) of EGFL7 using plasmid vector were named si-PANC-1, while transfected with negative control plasmid vector were called NC-PANC-1. Transwell assay was used to analyze the migration of PANC-1 cells. Real-time PCR and western blotting were used to detect the expression change of EGFL7 gene, EMT markers like E-Cadherin, N-Cadherin, Vimentin, Fibronectin and transcription factors like snail, slug in PANC-1, NCPANC-1, and si-PANC-1 cells, respectively. Results: After successful plasmid transfection, EGFL7 gene were dramatically knock-down by RNA interference in si-PANC-1 group. Meanwhile, migration ability decreased significantly, compared with PANC-1 and NC-PANC-1 group. Meanwhile, the expression of epithelial phenotype marker E-Cadherin increased and that of mesenchymal phenotype markers N-Cadherin, Vimentin, Fibronectin dramatically decreased in si-PANC-1 group, indicating a reversion of EMT. Also, transcription factors snail and slug decreased significantly after RNA interference. Conclusions: Current study suggested that highly-expressed EGFL7 promotes migration of PANC-1 cells and acts through transcription factors snail and slug to induce EMT, and further study is needed to confirm this issue.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.93-93
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• 2017

To develop rice cultivars with increased biomass and grain yield, superior lodging resistance is an essential trait. The new breeding approach can be adopted for the improvement of stem lodging resistance by enhancing culm strength. The resistance to breaking type lodging is attributed to bending moment of basal culm (M), which is composed of the section modulus (SM) and bending stress (BS). The resistance to the bending type lodging is attributed to flexural rigidity (FR) of stem, which is composed of the secondary moment of inertia (SMI) and Young's modulus (YM). Starch and cell wall components such as cellulose, hemicellulose and lignin also play a significant role in physical strength of culm, and thus affect lodging. Leaf Star has a superior lodging resistance due to its thick and stiff culm because of its high M and FR compared with Koshihikari. Furthermore, Leaf Star contains high densities of hemicellulose, cellulose and low lignin density in culm compared with Koshihikari. In this study, we performed QTL analysis for these traits associated with culm strength, using 94 recombinant inbred lines (RILs, $F_8$), derived from a cross between Leaf Star and Koshihikari. The SM in the RILs showed a continuous distribution. QTLs for SM were detected on chrs.2, 3 and 10. Leaf Star alleles increased SM on chrs. 2 and 3, but Koshihikari allele increased on chr.10. These QTLs overlapped with those QTLs identified using backcrossed inbred line derived from a cross between Chugoku 117 and Koshihikari, the parents of Leaf Star. The FR in Leaf Star was higher than that in Koshihikari due to the larger SMI and YM. 3 QTLs for SMI were detected on chrs.2, 3 and 10. Leaf Star alleles increased SMI on chrs.2 and 3, and Koshihikari alleles increased on chr.10. One QTL on chr.3 and two QTLs on chr.5 for hollocelulose content were detected with Leaf Star alleles contribution. Moreover, two QTLs were detected for hemicellulose density on chrs.3 and 5. Leaf Star allele increased hemicellulose density on chr.5, and Koshihikari allele increased on chr.3. Furthermore, two QTLs for cellulose density were detected on chr.5, and one QTL on chr.2. For starch content, one QTL on chr.3 and two QTLs on chr.5 with Leaf Star alleles contribution were detected. TULK-6 carrying a chromosome segment of Leaf Star on chr.5 in the Koshihikari genetic background showed higher densities of starch and hemicellulose than those in Koshihikari. These results suggest that the detected QTLs for culm strength could be utilized for the improvement of lodging resistance in rice by marker-assisted selection.

• 한국수정란이식학회지
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• 제19권3호
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• pp.291-299
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• 2004

요크셔종과 버크셔종 교배 실험 집단을 활용하여 양적형질 유전자좌 (QTL)의 발현 특성 관련 유전 양식을 조사하였다. 총 512두의 F$_2$ 자손이 F$_1$간의 65교배 조합으로부터 생산되었으며 표현형 조사 기록은 일당증제량(ADG), 평균 등지방 두께(ABF), 10번째 등뼈 부위 등지방 두께(TRF) 및 등심단면적(LEA), 최후 척추부위 등지방 두께 (LRF)였다. 125종의 유전자 표지 (microsatellite)에 대한 3세대 개체별 유전자형이 분석되었으며 이들 정보를 통하여 최소자승 회귀 모형을 이용한 interval mapping 방법을 적용하였다. QTL의 유전양식 여부 검정에 대한 절차를 도식화하기 위해 귀무가설인 통상의 벤델리안 모형에 근거를 두고 수행하였다. 경험적 다중 검정 통계량에 대한 임계치는 단일 개개의 염색체 수준과 게놈 전반에 걸친 실험수준으로 유도하였으며, permutation에 의해 유도된 임계치의 유효성을 검증하기 위해 본 연구에 활용된 실험축 집단 구조와 유사한 simulation 집단 구조에 의해 산출된 결과들과 비교하여 유효성이 인정되었다. 본 연구에 활용된 실험축 집단구조와 Genome 전반에 걸친 QTL imprinting 여부를 조사한 결과 13종의 QTL 에 대한 imprinting이 확인되었으며 이들 중 9종의 QTL 유전 양식은 부계로부터 전달된 자손에게만 발현되는 것으로 추론되었다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5231-5235
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• 2013

Objective: NF-E2-related factor 2 (Nrf2) is activated in several human malignancies. However, the role of Nrf2 in gastric cancer (GC) remains incompletely understood. In this study, we therefore analyzed associations of Nrf2 expression status with clinical features and chemotherapeutic resistance in GC. Materials and Methods: A total of 186 samples from GC patients who underwent gastrectomy were used for prognostic assessment. A further 142 samples from GC cases who received first-line combination chemotherapy were applied for investigation of chemoresistance. The Nrf2 expression was evaluated by immunohistochemistry in GC samples, and its relationship with clinicopathological parameters and chemotherapy sensitivity was analyzed. The effect of Nrf2 gene silencing on chemotherapy resistance was also examined by cell viability assay in vivo. Results: Of the 186 patients with GC, 104/186 (55.9%) showed high expression for Nrf2. The overexpression of Nrf2 was an independent predictor of overall survival [OS, hazard ratio (HR) 3.9; P=0.011] and disease-free survival (DFS, HR 4.3; P=0.002). The gene silencing of Nrf2 reduced resistance to cell death induced by 5-FU in GC cell lines. Conclusion: Our data show that Nrf2 is an independent prognostic factor in GC. Furthermore, Nrf2 confers resistance to chemotherapeutic drug 5-FU in GC cells. Taken together, Nrf2 is a potential prognostic marker and predictive for 5-FU resistance in GC.

• 한국임상수의학회지
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• 제11권2호
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• pp.529-537
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• 1994

Sector scanner which has a conical end is used to image through the intercostal space because heart is protected by the ribs. Cardiac data published all around the world were also obtained by sector scanner. Although scanners being used in every small animal practice and animal hospital at college in Korea include convex ape and linear type, linear type is not appropriate f3r cardiac scan because of a wide contact surface. The purpose of this study is to establish ultrasonographic images of normal cardiac structures by measuring shape, size of reflectable cardiac structure according to restraint position in scanning normal heart of the puppies with 6.5 MHz convex scanner(SonoAce 4500, Medison, Korea) used in our veterinary teaching hospital, Seoul national university. Seventeen male and female puppies considered having healthy hear by X-ray and clinical examination are used feom April to July 1994. Scanning point selection of probe head and the distinction of imaged cardiac structures were accomplished by necropsy and cardiac scanning performed through thoracotomy under general anesthesia. At 10 o'clock position of transducer(at an angle of 30$^{\circ}$ between imaginary line from elbow joint to 3rd sternum and probe head, 60$^{\circ}$ from body surface, 4th intercostal space of right thorax) with the marker of scanner toward the head of dogs right atrium, left atrium and left ventricle were observed in 2, 3, 4, 5 intercostal space(2cm from the sternum) of experimental dog positioned ventrodorsally under general anesthesia. Under these conditions, the numerical values of imaged diastolic hear are as follows : the distance from skin to apex(mean$\pm$S.D) 47.53$\pm$6.94mm, thickness of left ventricular wall 6.00$\pm$1.60mm, length of left ventricle 16.27$\pm$5.31mm, width of left ventricle 15,33$\pm$4.25mm, length of left atrium 12.33$\pm$3.82mm, width of left atrium 11. 33$\pm$3.94mm, length of right atrium 1.00$\pm$2.41mm, width of right atrium 11.21$\pm$2.76mm and the area of left ventricle 270.92$\pm$109.81mm$^2$, area of left atrium 98.00$\pm$41.08mm$^2$, area of right atrium 62.75$\pm$21.04mm$^2$.

• Journal of Plant Biotechnology
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• 제43권3호
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• pp.317-331
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• 2016

Development of disease resistant plant is one of the important objectives in rice breeding programs because biotic stresses can adversely affect rice growth and yield losses. This study was conducted to identify lines with multiple-resistance genes to biotic stress among 173 hybrid rice breeding lines and germplasms using DNA-based markers. Our results showed that one hybrid rice line [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66)] possessed 5 bacterial blight resistance genes (Xa4, xa5, Xa7, Xa13 and Xa21) while two hybrid rice lines [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66) and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessed 3 bacterial blight resistance genes (Xa4, Xa7 and Xa21, and Xa3, Xa4 and xa5). Molecular survey on rice blast disease revealed that most of these lines had two different resistant genes. Only 11 lines possessed Pib, Pi-5, and Pi-ta. In addition, we further surveyed the distribution of insect resistant genes, such as Bph1, Bph18(t), and Wbph. Three hybrid breeding lines [IR98161-2-1-1-k1-3 (IR86409-3-1-1-1-1-1/IRBB66), IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), and 7292s (IR75589-31-27-8-33S(S1) /IR102758B)] contained all three resistance genes. Finally, we obtained four hybrid rice breeding lines and germplasms [IR98161-2-1-1-k1-2 (IR86409-3-1-1-1-1-1/IRBB66), Damm-Noeub Khmau, 7290s, and 7292s (IR75589-31-27-8-33S(S1)/IR102758B)] possessing six-gene combination. They are expected to provide higher level of multiple resistance to biotic stress. This study is important for genotyping hybrid rice with resistance to diverse diseases and pests. Results obtained in this study suggest that identification of pyramided resistance genes is very important for screening hybrid rice breeding lines and germplasms accurately for disease and pest resistance. We will expand their cultivation safely through bioassays against diseases, pests, and disaster in its main export countries.

• Asian-Australasian Journal of Animal Sciences
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• 제30권7호
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• pp.985-993
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• 2017

Objective: The objective of this study was to evaluate increasing doses of a novel microbial phytase (Cibenza Phytaverse, Novus International, St. Charles, MO, USA) on standardized total tract digestibility (STTD) of P in canola meal (CM), corn, corn-derived distiller's dried grains with solubles (DDGS), rice bran (RB), sorghum, soybean meal (SBM), sunflower meal (SFM), and wheat. Methods: Two cohorts of 36 pigs each (initial body weight = $78.5{\pm}3.7kg$) were randomly assigned to 2 rooms, each housing 36 pigs, and then allotted to 6 diets with 6 replicates per diet in a randomized complete block design. Test ingredient was the only dietary source of P and diets contained 6 concentrations of phytase (0, 125, 250, 500, 1,000, or 2,000 phytase units [FTU]/kg) with 0.4% of $TiO_2$ as a digestibility marker. Feeding schedule for each ingredient was 5 d acclimation, 5 d fecal collection, and 4 d washout. The STTD of P increased (linear or exponential $p{\leq}0.001$) with the inclusion of phytase for all ingredients. Results: Basal STTD of P was 37.6% for CM, 37.6% for corn, 68.6% for DDGS, 10.3% for RB, 41.2% for sorghum, 36.7% for SBM, 26.2% for SFM, and 55.1% for wheat. The efficiency of this novel phytase to hydrolyze phytate is best described with a broken-line model for corn, an exponential model for CM, RB, SBM, SFM, and wheat, and a linear model for DDGS and sorghum. Based on best-fit model the phytase dose (FTU/kg) needed for highest STTD of P (%), respectively, was 735 for 64.3% in CM, 550 for 69.4% in corn, 160 for 55.5% in SBM, 1,219 for 57.8% in SFM, and 881 for 64.0% in wheat, whereas a maximum response was not obtained for sorghum, DDGS and RB within the evaluated phytase range of 0 to 2,000 FTU/kg. These differences in the phytase concentration needed to maximize the STTD of P clearly indicate that the enzyme does not have the same hydrolysis efficiency among the evaluated ingredients. Conclusion: Variations in enzyme efficacy to release P from phytate in various feedstuffs need to be taken into consideration when determining the matrix value for phytase in a mixed diet, which likely depends on the type and inclusion concentration of ingredients used in mixed diets for pigs. The use of a fixed P matrix value across different diet types for a given phytase concentration is discouraged as it may result in inaccurate diet formulation.

• Journal of Oral Medicine and Pain
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• 제26권1호
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• pp.39-50
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• 2001

In order to investigate the effect of Transforming growth factor ${\beta}1$(below TGF-${\beta}1$) and osteogenic protein-1(below Op-1) onto the myogenic differentiation, C2C12 satellite myoblastic cell line was cultured and treated with both growth factors. At first morphological changes with microscopical examination were examined, and isolated total RNA to analyse mRNA expression of bone marker proteins, muscle regulatory proteins, TGF-${\beta}$ receptor and their ligands by Northern blot analysis. And cellular proliferative inducibility of both growth factors was also tested to C2C12 cells. Incubating the cell with $5ng/m{\ell}$ of TGF-${\beta}1$ until 4 days almost inhibited multinucleated myotube formation expressing muscular regulatory proteins, and induced decreasing Id proteins. However, no osteoblastic phenotypes was induced by TGF-${\beta}1$ in C2C12 cells. The mRNA expression of TGF-${\beta}$ receptors with TGF-${\beta}1$ was conversed after 48 hours cultured. Type I TGF-${\beta}$ receptor was seemed to play a role in negative signalling for inhibition of myogenic differentiation. OP-1 dose dependently induced ALP activity, osteopontine production and bone sialoprotein production at concentrations above $100ng/m{\ell}$ and osteocalcin production at concentrations above $300ng/m{\ell}$. The concentration of OP-1 required to induce these osteoblastic phenotypes was the same as that required to almost completely inhibit myotube formation. Incubation with above $100ng/m{\ell}$ OP-1 suppressed the expression of mRNA for muscular egulatory proteins from 2 days after incubation. Expression of Id-1, 2, 3 mRNA were stimulated by OP-1 at concentration above $300ng/m{\ell}$. When C2C12 cells were treated with both growth factors, TGF-${\beta}1$ potentiated the inhibitory effect of OP-1 on myotube formation and expression of mRNA for myogenin at 12 days. And TGF-${\beta}1$ reduced osteocalcin and bone sialoprotein production induced by OP-1 at 12 days in C2C12 cells. Both growth factor had no mitogenic effect. These results indicate that OP-1 converts the differentiation pathway of C2C12 myoblasts into that of osteoblastic lineage cells and it's not heritable, but TGF-${\beta}1$ does not and has reversible inhibitory activity on the myogenic differentiation. TGF-${\beta}1$ and OP-1 play a role in myogenic differentiation via different mechanism between them.