• Biomolecules & Therapeutics
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• v.23 no.1
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• pp.19-25
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• 2015

Vascular endothelial growth factor (VEGF) is an important regulator of neovascularization. Hypoxia inducible nitric oxide (NO) enhanced the expression of VEGF and thymosin beta-4 ($T{\beta}4$), actin sequestering protein. Here, we investigated whether NO-mediated VEGF expression could be regulated by $T{\beta}4$ expression in HeLa cervical cancer cells. Hypoxia inducible NO production and VEGF expression were reduced by small interference (si) RNA of $T{\beta}4$. Hypoxia response element (HRE)-luciferase activity and VEGF expression were increased by the treatment with N-(${\beta}$-D-Glucopyranosyl)-N2-acetyl-S-nitroso-D, L-penicillaminamide (SNAP-1), to generate NO, which was inhibited by the inhibition of $T{\beta}4$ expression with $T{\beta}4$-siRNA. In hypoxic condition, HRE-luciferase activity and VEGF expression were inhibited by the treatment with $N^G$-monomethyl-L-arginine (L-NMMA), an inhibitor to nitric oxide synthase (NOS), which is accompanied with a decrease in $T{\beta}4$ expression. VEGF expression inhibited by L-NMMA treatment was restored by the transfection with pCMV-$T{\beta}4$ plasmids for $T{\beta}4$ overexpression. Taken together, these results suggest that $T{\beta}4$ could be a regulator for the expression of VEGF via the maintenance of NOS activity.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.11
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• pp.1746-1753
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• 2007

Steers (n = 335) of known genetic background from four fundamentally different growth types were subjected to two production systems to study differences in cooking loss (CL), tenderness, and chemical composition. Growth types were animals with genetic potential for large mature weight-late maturing (LL), intermediate mature weight-late maturing (IL), intermediate mature weight -early maturing (IE), and small mature weight-early maturing (SE). Each year, in a nine-year study, calves of each growth type were weaned and five steers of each growth type were developed on pasture or feedlot and harvested at approximately 20 and 14 mo of age, respectively. Data collected were CL and Warner-Bratzler shear force (WBS) for the Longissimus dorsi (LM), Psoas major (PS), and Quadriceps femoris (QF) muscles. Chemical composition was also determined from the right fore- and hindquarter. Data were analyzed using least squares analysis of variance for unequal subclass numbers. The beef growth $type{\times}production$ system interaction was significant for CL and WBS of the LM and ash in the lean trim of the forequarter. Growth types of LL and IL had greater (p<0.05) mean percentage CL in the PS and QF muscles than did IE and SE steers. Growth type LL had the highest (p<0.05) mean for both moisture and protein in the fore- and hindquarters; while SE had the lowest numerical mean value for moisture and protein in the fore- and hindquarters. Shear force of the PS did not differ (p>0.05) among steers of the four growth types. Increasing challenges to the cattle feeding industry may dictate that pasture development play a larger role in future production regimes. Producers should strive to match genetic growth type with available resources in order to remain viable and continue producing a quality product.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.4
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• pp.582-589
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• 2008

This study attempted to determine effects of recombinant porcine epidermal growth factor (pEGF) and glutamine (Gln) supplement on the growth performance and intestinal development of piglets weaned at 14 days of age. A total of ninety-six piglets were allotted to one of four dietary treatments which comprised inclusion of 1.0 mg pEGF supernatant/kg diet or 0.5% Gln both alone and in combination. Each treatment consisted of four replicates with six pigs per pen for a 28 days experimental period. Two pigs per replicate were sacrificed and gastrointestinal tract samples were collected on day 14. Data showed that dietary treatment failed to promote growth performance. On day 14, diets supplemented with pEGF elevated pancreatic chymotrypsin, jejunal alkaline phosphatase, sucrase, lactase and maltase activities (p<0.05), but failed to alter the small intestinal villus morphology, DNA, or protein content of gastrointestinal mucosa. Diets supplemented with Gln increased pancreatic chymotrypsin activity, tended to enhance the protein contents of gastric (p = 0.08) and jejunal mucosa (p = 0.09) but did not influence the serum IgA level or the enzyme activity in the gastrointestinal tract. On day 28, the diets supplemented with Gln increasedt (p<0.05) serum IgA and the proliferation of peripheral blood mononuclear cells by PHA stimulation. However, a combination of pEGF and Gln did not have a synergistic effect on these biomarkers in early-weaned piglets. The results demonstrate that diets supplemented with recombinant pEGF supernatant indeed improve intestinal digestive enzyme activity and diets supplemented with Gln increases the immune response in early-weaned piglets.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.11
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• pp.1529-1540
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• 2011

Silencing of a specific gene using RNAi (RNA interference) is a valuable tool for functional analysis of a target gene. However, information on RNAi for analysis of gene function in farm animals is relatively nil. In the present study, we have validated the interfering effects of siRNA (small interfering RNA) using both quantitative and qualitative gene silencing in buffalo granulosa cells. Qualitative gene knockdown was validated using a fluorescent vector, enhanced green fluorescence protein (EGFP) and fluorescently labeled siRNA (Cy3) duplex. While quantitatively, siRNA targeted against the luciferase and CYP19 mRNA was used to validate the technique. CYP19 gene, a candidate fertility gene, was selected as a model to demonstrate the technique optimization. However, to sustain the expression of CYP19 gene in culture conditions using serum is difficult because granulosa cells have the tendency to luteinize in presence of serum. Therefore, serum free culture conditions were optimized for transfection and were found to be more suitable for the maintenance of CYP19 gene transcripts in comparison to culture conditions with serum. Decline in fluorescence intensity of green fluorescent protein (EGFP) was observed following co-transfection with plasmid generating siRNA targeted against EGFP gene. Quantitative decrease in luminescence was seen when co-transfected with siRNA against the luciferase gene. A significant suppressive effect on the mRNA levels of CYP19 gene at 100 nM siRNA concentration was observed. Also, measurement of estradiol levels using ELISA (enzyme-linked immunosorbent assay) showed a significant decline in comparison to control. In conclusion, the present study validated gene silencing using RNAi in cultured buffalo granulosa cells which can be used as an effective tool for functional analysis of target genes.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.11
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• pp.5409-5413
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• 2012

At present, multiple myeloma (MM) remains an incurable disease and cologenic cells may be responsible for disease relapse. It has been proposed that CD20+/CD138- NCI-H929 cells could be hallmarks of MM clonogenic cells. Here, the immunology phenotype of NCI-H929 cells is described. Only a small population of CD20+/CD138- cells (<1%) was found in the NCI-H929 cell line, but CD20+/CD138- cells were not detected. We found that CD20+/CD138+ cells were able to exhibit cologenic capacity by colony formation assay and continuous passage culture. Proteins were analyzed by 1D-SDS-PAGE and TMT based quantitative differential liquid chromatography tandem mass spectrometry (LC-MS/MS). 1,082 non-redundant proteins were identified, 658 of which were differentially expressed with at least a 1.5-fold difference. 205 proteins in CD20+ cells were expressed at higher levels and 453 proteins were at lower levels compared with CD20- cells. Most proteins had catalytic and binding activity and mainly participated in metabolic processes, cell communication and molecular transport. These results proved that there are different biological features and protein expression profile between CD20+ and CD20- cells in the NCI-H929 cell line.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.9
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• pp.4435-4439
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• 2012

Purpose: This study aimed to explore the role of the Twist gene in the epithelial-mesenchymal transition of ovarian cancer. Methods: An RNA interference plasmid expressing a small interfering RNA (siRNA)-targeting Twist (Twist siRNA vector) was designed, constructed, and transfected into the human ovarian cancer cell line A2780. Transfection efficiency was assessed under a fluorescence microscope. Changes in the expression of Twist mRNA in A2780 after transfection with the pGenesil Twist shRNA plasmid were analyzed through RT-PCR. MTT assays and adhesion experiments were applied to determine changes in proliferation and adhesion ability of A2870 after transfection with the Twist shRNA plasmid. Changes in the expression of the E-cadherin and N-cadherin proteins in A2780 after transfection with the Twist shRNA plasmid were analyzed using Western blotting. Result: The restructuring plasmid pGenesil-Twist shRNA was constructed successfully. After 48 h of culture, 80% of the cells expressed high-intensity GFP fluorescence and stability. The expression of Twist decreased significantly after the transfection of the Twist shRNA plasmid (P<0.05). Proliferation of the transfected Twist shRNA cells showed no difference with that of the A2780-nontransfection or A2780-si-control groups (P>0.05) but the adhesion ability of A2780 decreased dramatically (P<0.05). Expression of the E-cadherin protein increased, whereas that of the N-cadherin protein decreased compared with that in the A2780-nontransfection or A2780-si-control groups (P<0.05). Conclusion: Twist is essential for epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.6
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• pp.3897-3901
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• 2013

Background: Optimal treatment for prostate cancer remains a challenge worldwide. Recently, T cell immunoglobulin mucin-3 (TIM-3) has been implicated in tumor biology but its contribution prostate cancer remains unclear. The aim of this study was to investigate the role of TIM-3 as a prognostic marker in patients with prostate cancer. Methods: TIM-3 protein expression was determined by immunohistochemistry and Western blotting in 137 prostate cancer tumor samples and paired adjacent benign tissue. We also performed cell proliferation assays using 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl- 2H tetrazolium bromide (MTT) and cell invasion assays. The effects of small interfering RNA (siRNA)-mediated knockdown of TIM-3 (TIM-3 siRNA) in two human prostate cancer cell lines were also evaluated. Results: TIM-3 expression was higher in prostate cancer tissue than in the adjacent benign tissue (P<0.001). High TIM-3 expression was an independent predictor of both recurrence-free survival and progression-free survival. TIM-3 protein was expressed in both prostate cancer cell lines and knockdown suppressed their proliferation and invasion capacity. Conclusions: TIM-3 expression is associated with a poor prognosis in prostate cancer. Taken together, our resutlts indicate that TIM-3 is a potential prognostic marker in prostate cancer.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.9
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• pp.1303-1313
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• 2017

Objective: This study evaluated the change and function of the pancreas, and small intestine in relation to growth performance of broilers on diets supplemented with raw soybean meal (RSBM) and protease. Samples of test ingredients and diets, after mixing and prior to being used were also assessed on contents of anti-nutritional factors. Methods: A $3{\times}3$ factorial study was used, with three levels of RSBM (commercial soybean meal [SBM] was replaced by RSBM at 0, 10%, or 20%) and protease (0.1, 0.2, or 0.3 g/kg). Each treatment was replicated six times with nine birds per replicate. Birds were housed in cages, in climate-controlled room and fed starter, grower and finisher diets. Results: Levels of trypsin inhibitors in the diets, containing varying levels of RSBM ranged between 1,730.5 and 9,913.2 trypsin inhibitor units/g DM. Neither RSBM nor protease supplementation in diets significantly affected (p>0.05) the body weight of broilers in the entire periods (0 to 35-d). Increasing the level of RSBM in diets increased the weight of the pancreas at d 10 (p<0.000), d 24 (p<0.001), and d 35 (p<0.05). Increasing levels of RSBM in the diets reduced the apparent ileal digestibility of crude protein (CP), and amino acid (AA) at d 24. Increasing level of RSBM in the diets decreased (p<0.01) pancreatic protein content, but this was increased (p<0.05) when protease was added to the diets (0 to 10-d). Increasing the level of protease improved the pancreatic digestive enzymes, including trypsin (p<0.05), chymotrypsin (p<0.01), and general proteolytic enzymes (p<0.05). Conclusion: The commercial SBM could be replaced at up to 20% by RSBM for broilers. Although protease supplementation slightly improved the digestive enzymes, and the ileal digestibilities of CP and AA, the CP and AA were negatively affected by increasing RSBM.

• Korean Journal of Plant Resources
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• v.31 no.3
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• pp.218-227
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• 2018

In this study, we investigated the effect of the extracts from Vaccinium oldhamii on cell proliferation and the regulatory mechanisms of cyclin D1 protein level in human cancer cells. The branch extracts from Vaccinium oldhamii (VOB) showed higher inhibitor effect against the cell growth than leave extracts (VOL) and fruit extracts (VOF) in human colorectal cancer, breast cancer, prostate cancer, non-small lung cancer, pancreatic cancer and liver cancer cells. In addition, VOB decreased cyclin D1 level at both protein and mRNA level. MG132 treatment attenuated VOB-mediated cyclin D1 downregulation. A point mutation of threonine-286 to alanine attenuated cyclin D1 degradation by VOB. In addition, the inhibition of nuclear export by leptomycin B (LMB) attenuated cyclin D1 degradation by VOB. But, the treatment of PD98059 (ERK1/2 inhibitor), SB203580 (p38 inhibitor), SP600125 (JNK inhibitor), LiCl ($GSK3{\beta}$ inhibitor), LY294002 (PI3K inhibitor) or BAY 11-7082 ($I{\kappa}K$ inhibitor) did not affect VOB-induced cyclin D1 degradation. In conclusion, VOB induced cyclin D1 degradation through redistribution of cyclin D1 from the nucleus to cytoplasm via T286 phosphorylation of cyclin D1, which resulted in the inhibition of cancer cell proliferation.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.48
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• pp.58-64
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• 2003

To enjoy a healthy life, it is important to have a well-balanced diet. However, in today's society, there is an increase in the consumption of preprocessed foods and frequency of eating out. Also the western diet, which is becoming move popular worldwide, contains relatively high levels of protein and fat, and a low amount of fiber, Furthermore, the increased availability of favorite foods has created a condition were the individual diet is less variable. With these conditions, it is difficult to maintain a diet that is nutritionally balanced. With these unbalanced diets, which are difficult to change, there has been an increase in adult disease and health problems, such as colon and breast cancer, It is speculated that metabolites for carcinogens are produced from diet components and that intestinal bacteria contribute to the production of these metabolites. Therefore, it is necessary to evaluate the relationships between health, diet, and intestinal microflora. Soybean oligosaccharide is composed of water-soluble saccharides that have been extracted from soybean whey, a by-product from the production of soy protein. This is mainly a mixture of mono-, di-, tri-, and tetrasac-charides, with the principle components being the oligosaccharide raffinose and stachyose. When consumed by humans, the oligosaccharides cannot be digested in the human duodenal and small intestinal mucosa, and these are selectively utilized by beneficial bifidobacteria in intestines. The results of acute and subacute toxicity tests, soy-bean oligosaccharides were nonpoisonous. Soybean oligosaccharides promote the growth of indigenous bifido-bacteria in the colon which by their antagonistic effects, suppress the activity of putrefactive bacteria. Also, they reduce toxic metabolites, detrimental enzymes and plasma lipid, and increase in the frequency of bowel evacuation and fecal quantities. Consequently, soybean oligosaccharides as functional foods components have potential roles in the prevention and medical treatment of chronic adult diseases. The study of processing property and physiological function of soybean oligosacchavides and development of high oligosaccharide variety allow the creation of new and exciting foodstuffs that aye functional healthy.