• Title/Summary/Keyword: 유전자 발현 변화

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Apoptotic Effect of co-treatment with HS-1200 and Cisplatin on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line (HS-1200과 cisplatin의 병용처리가 사람구강암세포에 미치는 세포자멸사 효과에 대한 연구)

  • Kim, Duk-Han;Kim, In-Ryoung;Park, Bong-Soo;Ahn, Yong-Woo;Jeong, Sung-Hee
    • Journal of Oral Medicine and Pain
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    • v.38 no.3
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    • pp.221-233
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    • 2013
  • Bile acids are polar derivatives of cholesterol essential for the absorption of dietary lipids and regulate the transcription of genes that control cholesterol homeostasis. Recently it have been identified the synthetic chenodeoxycholic acid (CDCA) derivatives HS-1200 and cisplatin showed apoptisis-inducing activity on various cancer cells in vivo and in vitro. This study was undertaken to investigate the synergistic apoptotic effect of co-treatment with HS-1200 and cisplatin on human tongue squamous cell carcinoma cells (SCC25 cells). To investigate whether the co-treatment with HS-1200 and cisplatin compared to each single treatment efficiently reduces the viability of SCC25 cells, MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis DNA hypoploidy. Westen blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) change were also assayed. In this study, co-treatment with HS-1200 and cisplatin on SCC25 cells showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, reduction of MMP and proteasome activity, the increase of Bax and the decrease of Bcl-2, decrease of DNA content, the release of cytochrome c into cytosol, translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD) whereas each single treated SCC25 cells did not show these patterns. Although the single treatment of $25{\mu}M$ HS-1200 and $4{\mu}g/ml$ cisplatin for 24 h did not induce apoptosis, the co-treatment of these reagents prominently induced apoptosis. Therefore our data provide the possibility that the combination therapy with HS-1200 and cisplatin could be considered as a novel therapeutic strategy for human squamous cell carcinoma.

Comparative Functional Analysis of the Malate Dehydrogenase(Mor2) during in vitro Maturation of the Mouse and Porcine Oocytes (체외성숙 과정 중 생쥐와 돼지 난자의 Malate Dehydrogenase(Mor2)의 기능에 대한 비교 분석)

  • Kim, Eun-Young;Kim, Kyeoung-Hwa;Kim, Yun-Sun;Lee, Hyun-Seo;Kim, Yu-Nna;Lee, Kyung-Ah
    • Development and Reproduction
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    • v.11 no.3
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    • pp.263-272
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    • 2007
  • Contrast to mouse where its in vitro maturation rates are high without specific supplements or presence of the cumulus cells, there are some species, such as porcine, where its in vitro oocyte maturation rates are still very low. This comparative study was conducted to investigate the role of malate dehydrogenase(Mor2) during oocyte maturation by RNAi in the mouse and porcine. The Mor2 double-stranded RNA(dsRNA) was prepared speciesspecifically and microinjected into the cytoplasm of denuded germinal vesicle(GV) oocytes. Oocytes were cultured for 48 h(porcine) and 16 h(mouse) in M199 with 10% porcine follicular fluid, pyruvate, p-FSH, EGF, cystein, and estradiol-$17{\beta}$. We measured changes in oocyte morphology, maturation rates and mRNA levels after Mor2 RNAi. We confirmed gene sequence-specific knock down of Mor2 mRNA in both species after Mor2 RNAi. In contrast to our previous finding that mMor2 RNAi resulted in GV arrest in the mouse, we found that pMor2 RNAi resulted in MI arrest in denuded porcine oocytes(58%), but developed to MII(84.4%) in COCs. To determine whether this difference between mouse and porcine RNAi is due to differences in culture media, we cultured mouse oocytes in the M199 media for 16 h after mMor2 RNAi. Mouse oocytes were developed to MII stage(62%) and there was no statistical difference compared to that of non-injected(76.8%) and buffer-injected(73.3%) control groups. Therefore, we concluded that the mouse and porcine oocytes are having different metabolic systems in relation to malate dehydrogenase for oocyte maturation. This could be a basis for differences in maturation rates in vitro in two species. Further scrutinized studies on the metabolic pathways would led us in finding better culture system to improve oocyte maturation rates in vitro, especially in more challenging species like the porcine.

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Effect of High-Fat Diet-induced Obesity on the Incidence and Progression of Prostate Cancer in C57BL/6N Mouse (C57BL/6N 마우스에서 전립선암의 발병률 및 진행에 대한 고지방식이-유도 비만의 영향)

  • Choi, Yun Ju;Kim, Ji Eun;Lee, Su Jin;Gong, Jeong Eun;Jin, Yu Jeong;Lee, Jae Ho;Lim, Yong;Hwang, Dae Youn
    • Journal of Life Science
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    • v.32 no.7
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    • pp.532-541
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    • 2022
  • Obesity induced by high-fat diet (HFD) is verified as a strong risk factor and negative prognostic factor for prostate cancer in several genetically engineered mice although it was not examined in the normal mice. To investigate whether HFD-induced obesity can affect the development and progression of cancer in the prostate of normal mice, alterations in the weight and histological structure of the prostate as well as the expression of cancer-related proteins were analyzed in obese C57BL/6N mice fed with 60% HFD for 16 weeks. First, HFD-induced obesity, including an increase in organ weight, body weight, fat accumulation, and serum lipid profile, was successfully induced in C57BL/6N mice after HFD treatment. The total weight of the prostate significantly increased HFD-induced obesity in the model mice compared with the control group. Among the four lobes of the prostate, the weight of the ventral prostate (VP) and anterior prostate (AP) were higher in HFD-induced obesity model mice than in the control group, although the weights of the lateral prostate (DLP) and seminal vesicle (SV) were constantly maintained. In addition, the incidences of hyperplasia and non-hodgkin's lymphoma (NHL) in the histological structure were remarkably increased in HFD-induced obesity model mice, while the epithelial thickness was higher in the same group. A significant increase in the phosphorylation levels of key proteins in the AKT (protein kinase B) signaling pathway was detected in HFD-induced obesity model mice. Therefore, these results suggest that HFD-induced obesity can promote hyperplasia and NHL in the prostates of C57BL/6N mice through the activation of the AKT signaling pathway.

Enhancement of Bioactive Compounds in Mugwort Grown under Hydroponic System by Sucrose Supply in a Nutrient Solution (양액 내 자당 처리에 의한 수경재배 쑥의 생리활성물질 증진)

  • Moon-Sun Yeom;Jun-Soo Lee;Myung-Min Oh
    • Journal of Bio-Environment Control
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    • v.32 no.1
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    • pp.23-33
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    • 2023
  • Sucrose (suc) is a disaccharide that consists of glucose (glu) and fructose (fru). It is a carbohydrate source that acts as a nutrient molecule and a molecular signal that regulates gene expression and alters metabolites. This study aimed to evaluate whether suc-specific signaling induces an increase in bioactive compounds by exogenous suc absorption via roots or whether other factors, such as osmotic stress or biotic stress, are involved. To compare the osmotic stress induced by suc treatment, 4-week-old cultured mugwort plants were subjected to Hoagland nutrient solution with 10 mM, 30 mM, and 50 mM of suc or mannitol (man) for 3 days. Shoot fresh weight in suc and man treatments was not significantly different from the control. Both man and suc treatments increased the content of bioactive compounds in mugwort, but they displayed different enhancement patterns compared to the suc treatments. Mugwort extract treated with suc 50 mM effectively protected HepG2 liver cells damaged by ethanol and t-BHP. To compare the biotic stress induced by suc treatment, 3-week-old mugwort plants were subjected to microorganism and/or suc 30 mM with Hoagland nutrient solution. Microorganisms and/or suc 30 mM treatments showed no difference about the shoot fresh weight. However, sugar content in mugwort treated with suc 30 mM and microorganism with suc 30 mM treatment was significantly higher than that of the control. Suc 30 mM and microorganism with suc 30 mM were effective in enhancing bioactive compounds than microorganism treatment. These results suggest that mugwort plants can absorb exogenous suc via roots and the enhancement of bioactive compounds by suc treatment may result not from osmotic stress or biotic stress because of microorganism, but by suc-specific signaling.

Study on the Genetic Variations of the Economic Traits by Backcrossing in Commercial Chickens (실용계군에 있어서 누진퇴교배에 의한 주요경제형질의 유전적 변이에 관한 연구)

  • 이종극;오봉국
    • Korean Journal of Poultry Science
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    • v.16 no.2
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    • pp.61-71
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    • 1989
  • The purposes of this study were to investigate the genetic variations by backcrossing in commercial chickens. Backcrossing was carried out successively back to parent stock (P.S). Heritabilities and genetic correlation coefficients were estimated to verify the genetic variations. The data obtained from a breeding programme with commercial chickens (I strain) were collected from 1955 to 1987 at Poultry Breeding Farm, Seoul National University. Data came from a total of 1230 female offspring. The results obtained are summarized as follows: 1. The general performance ($Mean\pmStandard deviation$) of each trait was $663.94\pm87.11$g for 8 weeks body weight, $1579.1\pm155.43$g for 20 weeks body weight, $2124.1\pm215.3$g for 40 weeks body weight, $2269.1\pm242.94$g for 60 weeks body weight, $168.43\pm12.94$ day for a9e at sexual maturity (SM), $214.52\pm29.82$ eggs , for total egg number to 60 weeks of age (TEN), $61.45\pm3.48$ g for average weight (AEW), $13180.7\pm1823.22$ g for total egg mass to 60 weeks of age(TEM). All traits, except 10 weeks body weight and AEW, were significant for the degrees of backcross (p<0.01). 2. The pooled estimates of heritabilities derived from the sire, dam and combined variance components were 0.47~0.52 for age at sexual maturity (SM), 0.07~0.37 for total egg number (TEN), 0.40~0.54 for average egg weight (AEW), 0.18~0.27 for total egg mass (TEM). High heritability estimates were found for SM and AEW. TEN and TEM were estimated to be a lowly heritable traits. Heritability estimates from dam components were higher than those from sire components. These differences might be due to non-additive genetic effect and maternal effect. 3. The estimates of heritabilities and standard errors derived from combined variance components for different degrees of backcross were $0.47\pm0.11$ (BCO), $0.42\pm0.16$ (BC1), $0.51\pm0.29$ (BC2) for TEN, $0.59\pm0.20$ (BCO), $0.43\pm0.17$ (BC1), $0.35\pm0.18$ (BC2) for AEW, $0.28\pm0.12$(BC0), $0.20\pm0.11$(BC1), $0.18\pm0.14$ (BC2) for TEM. Heritability estimates for AEW and TEM were decreased by backcrossing while those for SM and TEN remained constant. Since backcrossing contributes to increased homozygosity, the genetic variation of the traits (AEW and TEM) decreased . 4. The pooled estimates of genetic correlation coefficients were -0.55 between SM and TEN, 0.20 between SM and AEW, -0.29 between TEN and AEW, 0.82 between TEM and TEN, 0.31 between TEM and AEW, -0.42 between TEM and SM. The genetic correlation between TEM and TEN was higher than that between TEM and AEW, and it was suggested that egg mass was strongly affected by egg number. Also, age at sexual maturity(SM) contributes to egg mass(TEM). 5. When backcrossing was carried out successively, the genetic correlation between TEM and TEN increased (BC0:0.79, BC1:0.82, BC2:0.91) but those between TEM and SM decreased (BC0:-0.54, BC1:-0.36, BC2:-0.09) with successive backcrosses.

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A Natural L-Arginine Analog, L-Canavanine-Induced Apoptosis is Suppressed by Protein Tyrosine Kinase p56lck in Human Acute Leukemia Jurkat T Cells (인체 급성백혈병 Jurkat T 세포에 있어서 L-canavanine에 의해 유도되는 세포자살기전에 미치는 단백질 티로신 키나아제 p56lck의 저해 효과)

  • Park, Hae-Sun;Jun, Do-Youn;Woo, Hyun-Ju;Rue, Seok-Woo;Kim, Sang-Kook;Kim, Kyung-Min;Park, Wan;Moon, Byung-Jo;Kim, Young-Ho
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1529-1537
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    • 2009
  • To elucidate further the antitumor effects of a natural L-arginine analogue, L-canavanine, the mechanism underlying apoptogenic activity of L-canavanine and its modulation by protein tyrosine kinase $p56^{lck}$ was investigated in human Jurkat T cells. When the cells were treated with 1.25 to 2.5 mM L-canavanine for 36 h, several apoptotic events including mitochondrial membrane potential (${\Delta\Psi}m$) loss, activation of caspase-9, -3, -8, and -7, poly (ADP-ribose) polymerase (PARP) degradation, and DNA fragmentation were induced without alteration in the levels of Fas or FasL. These apoptotic changes were more significant in $p56^{lck}$-deficient Jurkat clone JCaM1.6 than in $p56^{lck}$-positive Jurkat clone E6.1. The L-canavanine-induced apoptosis observed in $p56^{lck}$-deficient JCaM1.6 cells was significantly reduced by introducing $p56^{lck}$ gene into JCaM1.6 cells by stable transfection. Treatment of JCaM1.6/lck cells with L-canavanine caused a transient 1.6-fold increase in the kinase activity of $p56^{lck}$. Both FADD-positive wild-type Jurkat T cell clone A3 and FADD-deficient Jurkat T cell clone I2.1 exhibited a similar susceptibility to the cytotoxicity of L-canavanine, excluding involvement of Fas/FasL system in triggering L-canavanine-induced apoptosis. The L-canavanine-induced apoptotic sub-$G_1$ peak and activation of caspase-3, -8, and -7 were abrogated by pan-caspase inhibitor (z-VAD-fmk), whereas L-canavanine-induced activation of caspase-9 was not affected. These results demonstrated that L-canavanine caused apoptosis of Jurkat T cells via the loss of ${\Delta\Psi}m$, and the activation of caspase-9, -3, -8, and -7, leading to PARP degradation, and that the $p56^{lck}$ kinase attenuated the ${\Delta\Psi}m$ loss and activation of caspases, and thus contributed as a negative regulator to L-canavanine-induced apoptosis.

Fermentation Properties and Increased Health Functionality of Kimchi by Kimchi Lactic Acid Bacteria Starters (김치 유산균 Starter를 이용한 김치의 발효 특성 및 기능성 증진 효과)

  • Bong, Yeon-Ju;Jeong, Ji-Kang;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.11
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    • pp.1717-1726
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    • 2013
  • Fermentation characteristics and health functionalities of kimchi by inoculating kimchi lactic acid bacteria (LAB) starters were studied. We manufactured single LAB starter kimchi (Lactobacillus plantarum pnuK, Lactobacillus plantarum 3099K, Leuconostoc mesenteroides pnuK), mixed LAB starter kimchi (Lb. plantarum pnu/Leu. mesenteroides pnuK, Lb. plantarum 3099/Leu. mesenteroides pnuK) with inoculum size of $10^6$ CFU/g, as well as naturally fermented kimchi (NK), and fermented them for 6 days at $15^{\circ}C$. The pH and acidity of the early phase of fermentation were not different, but kimchi with the starters showed rapid changes in the pH and acidity from 2 days of fermentation. As the fermentation progressed, the level of total aerobic bacteria and Lactobacillus sp. increased similarly with or without Lb. plantarum (LP) inoculation. However, the level of Leuconostoc sp. was high in kimchi inoculated with Leuconostoc sp. starter. In the sensory evaluation test, kimchi with starters received higher overall acceptability scores than those of NK; mixed starter added kimchi earned the highest score. In DPPH and hydroxyl radical scavenging activity, kimchi with the starters exhibited higher activity than that of NK. In the MTT assay of HCT-116 and HT-29 human colon cancer cells, NK showed inhibition rates of 63.4 and 51.9%, but LPpnuK achieved 77.1 and 68.8%, respectively. This study showed that inoculating starters in kimchi increased in vitro antioxidant and anticancer activities, and single starter (LP) added kimchi revealed higher functionality than the kimchi with mixed starter. Kimchis with the starters effectively up-regulated the gene expressions of the pro-apoptotic gene of Bax, but down-regulated Bcl-2. They promoted expressions of p53 and p21, and suppressed expressions of inflammation-related genes, iNOS and COX-2, compared with NK. Taken together, it is expected that using starters may help manufacture kimchi with improved sensory quality and health functionality.

Evaluation of Scab Resistance and Effect of Photosynthetic Rates on Fruit Characteristics among Elite Pear Seedlings (배 우량계통의 검은별무늬병 저항성 평가 및 광합성률이 과실 품질에 미치는 영향 구명)

  • Won, Kyung-Ho;Kang, Sam-Seok;Kim, Yoon-Kyeong;Sherzod, Rajametov;Lim, Kyeong-Ho;Lee, Han-Chan
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.117-122
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    • 2013
  • BACKGROUND: The scab, which is caused by Venturia nashicola, gives serious damages to pear trees. 'Niitaka' accounts for 82% of areas in pear cultivation. However 'Niitaka' is a scab susceptible cultivar. So, most of Korean farmers who growing pear trees have suffered by economic losses with the scab. In this research, we evaluated the scab resistance among elite pear seedlings to clarify genetics about the scab resistance. And we analyzed photosynthetic features with these seedlings to develop suitable cultivar which is advantageous for producing quality fruits during the growth and development of plants. METHODS AND RESULTS: We measured the rates of scab incidence among seedlings in a field experiment condition and an in-vitro test. An in-vitro test has been done with field experiment-based results. We made plant materials by grafting branches of each seedlings with 'Kongbae' rootstocks. And they had been grown for one month. Then, scab conidia suspension is sprayed to seedlings and sustained for 40 days under the controlled environment. As the results, 6 seedlings displayed lower incidence rates than other seedlings and 'Niitaka'. We also measured instant photosynthetic rates of each seedlings to determine the correlation between photosynthetic rates and fruit characteristics. However, it seemed that there is no correlation between them. CONCLUSION(S): Among the seedlings, 6 seedlings displayed the higher resistance to scab than other seedlings and 'Niitaka'. This characteristics is considered to be come from the gene expression of European pear. And we found that photosynthetic rate in trees rarely does not influence the fruit characteristics. It is considered to be affected by cultivar's own characteristics.

In vitro and in vivo Effects of Extracts of Lentinus edodes on Tumor Growth in a Human Papillomavirus 16 Oncogenes-transformed Animal Tumor Model -Apoptosis-mediated Tumor Cell Growth Inhibition- (자궁경부암동물세포에서 표고버섯의 in vitro 및 in vivo 항암효과 -Apoptosis에 의한 종양세포주의 성장억제-)

  • Park, Jeong-Min;Lee, Sung-Hyun;Kim, Jung-Ok;Park, Hong-Ju;Park, Jae-Bok;Sin, Jeong-Im
    • Korean Journal of Food Science and Technology
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    • v.36 no.1
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    • pp.141-146
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    • 2004
  • Fungal products indirectly mediate anti-tumor effects in vitro and in vivo. To investigate whether Lentinus edodes might possess direct anti-tumor substance, L. edodes was extracted and tested on human papillomavirus (HPV) 16 oncogenes-associated animal tumor cells (TC-1) and in an animal tumor model. Only water extract displayed direct anti-proliferative effects in TC-1 tumor cells in vitro. This inhibition was dose-dependent, and inhibitory concentration ($IC_{50}$) was $800\;{\mu}g/mL$. Fungal extracts also showed growth inhibition to human cervical cancer cells (CaSki and HeLa) similarly to TC-1 tumor cells. When fungal extracts were added at a high dose (1.5 mg/mL), cell growth was inhibited within 6 hr following extract treatment. Cell growth inhibition was blocked by heat treatment, but not by low pH, which is indicative of heat sensitivity of this anti-proliferative substance. Cell growth suppression was mediated by apoptosis, as determined by Annexin V and propidium iodide staining. When challenged with TC-1 cells, direct intratumoral injection of fungal extracts resulted in some positive effect on tumor growth inhibition, as compared to oral delivery. Results suggest that heat labile substance of L. edodes suppresses growth of HPV oncogenes-associated tumor cells through apoptosis.

Usefulness of Low Dose Oral Contrast Media in $^{18}F-FDG$ PET/CT ($^{18}F-FDG$ PET/CT에서 저용량 경구용 조영제의 유용성)

  • An Young-Sil;Yoon Joon-Kee;Hong Seon Pyo;Joh Chul-Woo;Yoon Seok-Nam
    • Nuclear Medicine and Molecular Imaging
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    • v.40 no.5
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    • pp.257-262
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    • 2006
  • Purpose: The standard protocol using large volume of oral contrast media may cause gastrointestinal discomfort and contrast-related artifacts in PET/CT. The aim of this study was to evaluate the usefulness of low dose oral contrast in $^{18}F-FDG$ PET/CT. Materials and Methods: We retrospectively reviewed the whole-body PET/CT images in a total of 435 patients. About 200 ml of oval contrast agent (barium sulfate) was administered immediately before injection of $^{18}F-FDG$. The FDG uptake of intestines was analyzed by visual and semi- quantitative method on transaxial, coronal and saggital planes. Results: Seventy (16%, 113 sites) of 435 images showed high FDG uptake (peak SUV > 4); 50 (74%, 84 sites) with diffuse and 20 (15%, 29 sites) with focal uptake. The most commonly delivered site of oral contrast media was small bowel (n=27, 39%). On PET/CT images, FDG uptake coexisted with oral contrast media in 26 patients (54%, 38 sites) with diffuse pattern and 9 (45%, 9 sites) with focal pattern, and by sites, those were 38 (45%) and 9 (31%), respectively. In small bowel regions, the proportion of coexistence reached as high as 61% (29/47 sites). A visual analysis of available non-attenuation corrected PET images of 27 matched regions revealed no contrast-related artifact. Conclusion: We concluded that the application of low dose contrast media could be helpful in the evaluation of abdominal uptake in the FDG PET/CT image.