• Title/Summary/Keyword: P19 cells

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Production and Characterization of Physiological Properties of Carotenoid from a Marine Bacterium Curtobacterium sp. (해양미생물로부터 Carotenoid의 생산 및 그 생리활성)

  • r김종덕;강동수;김민용;최명락;임현수;백승한;서효진;김대현;공재열
    • KSBB Journal
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    • v.15 no.6
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    • pp.621-629
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    • 2000
  • A marine bacterium producing carotenoid was isolated from the Yosu coastal area of South Korea, and has been recorded as MCPBK-1. It was identified as Curtobacterium sp.. The optimum conditions of marine carotenoid fermentation from Cutobacterium sp. were pH 7.0, a temperature of $25^{\circ}C$, 4 mM fructose as a carbon source, 0.07% tryptone as a nitrogen source, 0.5 mM $M^{+2}$ ion as a mineral source and $1{\;}\mu\textrm{M}$ of cyanocobalamine as a growth factor in a $7{\;}\ell$ jar-fermentor. 13.0 mg/ml of the marine carotenoid were produced under optimum conditions. The crude marine carotenoid isolated was composed of 5 different compounds, i.e : tunaxanthin(86.6%), diatoxanthin (7.1%), ${\beta}-carotene$ (2.1%), canthaxanthin(1.9%) and cynthiaxanthin (1.9%). Physiological properties including antibacterial activity, cytotoxic effect, antioxidative effect and free radical scavenging activity were characterized with the crude carotenoid, which exhibited no antibacterial activity against E. coli and Lactobacillus bulgaricus, but a strong cytotoxic effect against cancer cells such as HepG2 (Hepatocellular carcinoma, human, ATCC HB-8065) and HeLa (Cervical carcinoma, human, ATCC CCL-2) cells, the ratios of impediment were 86.4% and 39.2%, respectively. This carotenoid, also, expressed a strong antioxidative effect (83%) against CCL-13 (diploid, monotypic hepatocyte, human, ATCC CCL-13) and exhibited free radical scavenging activity (43.4%) when using at a concentration of $50{\;}\mu\textrm{g}/ml$ of the crude carotenoid.

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Effects of various Herbal Acupuncture on Adjuvant Arthritis in Rats (다양한 약침제제(藥鍼製劑)가 Rat의 Adjuvant 관절염(關節炎)에 미치는 영향(影響))

  • Ha, Jee-Yong;Lee, Seon-Goo;Yu, Byeong-Gil
    • Journal of Pharmacopuncture
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    • v.3 no.2
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    • pp.169-189
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    • 2000
  • To investigate effects of vanous herbal-acupuncture, which were Daebangpungtang, Oyaksungisan, Dokhwalgisaengtang, and Binsosan on Adjuvant Arthritis in rats, the edema rate, the number of WBC, the quantity of TNF-${\alpa}$, COX -2, and IL-6, and histological test of the muscular tissue were measured in the arthritis part. After elicilating arthritis of Sprague Dawely(SD) rats by injection of Freund's complets adjuvant for 2 weeks, saline was injected for the control group, Daebangpungtang acua-acupuncture was injected for the Daebangpungtang group, Oyaksungisan acua-acupuncture was injected for the Oyaksungisan group, Dokhwalgisaengtang acua-acupuncture was injected for the Dokhwalgisaengtang group, and Binsosan acua-acupuncture was injected for the Binsosan group during 30days. Selected point was ST35 in all groups. 1. The number of WBC was $8.24{\pm}0.51$($10^3$/ml) in the normal group and $27.35{\pm}3.51$($10^3$/ml) in the control group. the Daebangpungtang group and the Oyaksungisan group decreased each $20.42{\pm}2.75$($10^3$/ml) and $19.78{\pm}4.99$($10^3$/ml) in the Exp.ll group. This fact showed that the group Exp.ll was more effective than the control group effectively. (P<0.05) 2. The volume of the paw were checked. The volume of paw was $0.62{\pm}0.11mm in the control group and $0.45{\pm}0.08$mm in the Daebangpungtang group and $0.47{\pm}0.07$mm in the Oyaksungisan group, the swelling of the paw was restricted significantly in the Daebangpungtang and Oyaksungisan group(P<0.05) 3. The bands of the TNF-${\alpa}$, COX -2 in the muscular tissue of the control group wcre certain and thick than other groups(except normal group), and the band of the IL-6 in the muscular tissue of all group(except normal group) were similar forms. 4. In histological finding, because of severe inflammatory reaction, remarkably irregular tissue and large amount of inflammatory cells were found in thc control and Dokhwalgisaengtang group. But the Oyaksungisan and Daebangpul1gtang group showed small amount of inflammatory cells, the refrained inflammatory state and even recovering state. From these results, it is showed Oyaksugisan and Daebangpungtang acua-acupuncturc refrain inflammatory reaction and muscular tissue necrosis in SD rats paw were induced by Freund's complete adjuvant

Cellular Iron Uptake from Aqueous Solutions depending on Reaction Conditions by genetically engineered Saccharomyces cerevisiae (재조합 Saccharomyces cerevisiae에 있어서 반응조건에 따른 수용성 철의 생체 흡수)

  • Kim Sang-Jun;Chang Yu-Jung;Park Chung-Ung;Jeong Yong-Seob;Kim Kyung-Suk
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.441-445
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    • 2004
  • Cellular iron uptake was performed in the yeast Saccharomyces cerevisiae that transformed with human ferritin H- and L-chain genes. The recombinant yeasts were enriched in YEP medium supplemented with $2\%$ galactose for 3 days and the iron uptake was followed by incubating the cells with iron in 20 mM MOPS buffer (pH 6.5). The reactions were examined under different conditions including the iron compounds of Fe(II) and Fe(III), the concentration of iron, the concentration of cells and the reaction time. From our results, the recombinant yeast YGH2 producing H-chain ferritin showed higher cellular iron concentration at the cell concentration of 100 mg/ml than 200 mg/ml. Iron presented as Fe(II) rather than Fe(III) was taken up more efficiently. Iron uptake increased slightly when iron was added up to 14.3 mM Fe(II) and then its cellular iron concentration was $16.7{\pm}0.7\;{\mu}mol/g$ cell wet wt. In addition, the iron uptake reaction reached to maximum at about 2 hr incubation.

Anti-tumor Effect of Combined Betacarotene with X-irradiation in the Mouse Fibrosarcoma : Cytotoxicity and Tumor Growth Delay (쥐 섬유육종에서 베타카로틴과 방사선조사 병용의 항종양 효과: 세포독성 및 종양성장 지연에 미치는 영향)

  • Kwon Hyoung-Cheol;Yang Moon-Sik
    • Radiation Oncology Journal
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    • v.18 no.2
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    • pp.133-137
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    • 2000
  • Purpose :To investigate whether combined beta-carotene with X-Irradiation has more enhanced radition response than X-irradiation or not, we peformed a experiment about in vitro cytotoxlcity of beta-carotene and/or X-irradiation in the fibrosarcoma cells, tumor growth delay of combined beta-caroten with/or X-irradiation in the mouse fibrosarcoma. Materials and Methods : 2$\%$ emulsion of beta-carotene was serially diluted and used. X-Irradiation was given by 6 MeV linear accelerator. The cytotoxicity of beta-carotene in vitro was evaluated from clonogenic assay. To compare the cytotoxiclty between combined beta-carotene with X-irradiation and X-irradiation group, 2 mg/ml of beta-carotene was contacted to fibrosarcoma (FSall) cells for 1 hour before X-irradiation. For the tumor growth delay, single 20 Gy was given to FSall tumor hearing C3H/N mice whic was classified as beta-crotene with X-irradiation group (n=5) and X-irradiation alone group (n=5). 0.2 ml of 20 mg/kg of beta-carotene were i.p. injected to mice 30 minute before X-irradiation in the beta-crotene with X-irradiation group. The tumor growth delay defined as the time which reach to 1,000 mm$^{3}$ of tumor volume. Results : (1) Cytotoxicity in vitro: 1) survival fraction at beta-carotene concentration of 0.002,0.02,0.2 and 2 mg/ml were 0.69$\pm$0.07, 0.59$\pm$0.08, 0.08$\pm$0.008 and 0.02$\pm$0.006, respectively. 2) each survival fraction at 2, 4, 6 and 8 Gy in the 2 mg/ml of beta-carotene + X-irradiation group were 0.13$\pm$0.05, 0.03$\pm$0.005, 0.01 $\pm$0.002 and 0.009$\pm$0.0008, respectively. But each survival fraction at same irradiation dose in the X-irradiation group were 0.66$\pm$0.05, 0.40$\pm$0.04, 0.11$\pm$0.01 and 0.03$\pm$0.006, respectively(p<0.05). (2) The time which reach to 1,000 mm$^{3}$ of tumor volume of beta-carotene + X-irradiation group and X-irradiation alone group were 18, 19 days, respectively(p>0.05) Conclusion : The contact of beta-caroten to Fsall cells showed mild cytotoxicity which 띤as increased according to concentration. The cytotoxicity of combined beta-carotene with X-irradiation more increased than that of X-irradiation, additionally, And there was significant difference of cytotoxicity between two groups. But there were no significant difference of the growth delay of fibrosarcoma between two groups.

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Production of Cloned Miniature Pig by Surrogate Mother Conditions (대리모의 준비 조건 변화를 통한 복제미니돼지의 생산)

  • Hur, Chang-Gi;Yang, Hae-Young;Lee, Eun-Kyeong;Han, Joo-Hee;Park, Chun-Gyu;Shin, Teak-Soon;Lee, Hong-Gu;Kang, Han-Seok;Ahn, Jong-Deok;Cho, Seong-Keun
    • Journal of Embryo Transfer
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    • v.27 no.1
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    • pp.1-7
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    • 2012
  • Somatic cell nuclear transfer (SCNT) for miniature pig has been developed for xenotransplantation and many other biomedical experiments. However, the efficiency of SCNT is still very low due to many factors. To optimize the surrogate mother condition for improvement of cloned miniature pigs efficiency, we investigated the effect of the status of surrogate mother on pregnancy, farrowed rate in SCNT pigs. After SCNT with mesenchymal stem cells as donor cells, the SCNT embryos were surgically transferred into the oviduct of surrogated pigs. To compare the effects of status of surrogate pigs on pregnancy, surrogate pigs were prepared by artificial abortion at day 20~29 (Group 1), 30~39 (Group 2), and 40~45 (Group 3) of gestation. After SCNT embryos transfer in three different status of surrogate pigs, Group 2 (56.3%) and 3 (55.6%) had significantly ($p$ <0.05) higher the pregnancy rate than group 1 (0%) at day 30 of gestation. The status of ovulation in surrogate pig also was investigated. Post-ovulation status (54.8%) had higher proportion than pre-ovulation status (38.7%) and ovulation status (6.5%). We obtained 19 cloned miniature piglets from seven surrogate gilts and five piglets are living healthy but fourteen piglets died soon after birth or stillbirth. The weights of piglets greatly differ from 254 to 1,296 g. Microsatellite analysis showed that cloned piglets were genetically different from the surrogate mother and cloned piglets were genetically equal to the donor cell. In conclusion, the present result indicates that artificially abortion method can improve the efficiency of pregnancy after SCNT in pigs. This study will provide available method for the further study and application in the field of xenotransplantation.

Increased in vitro Anticancer Effects of Potassium Bamboo Salt (칼륨죽염의 in vitro 항암 기능성 증진 효과)

  • Zhao, Xin;Jeong, Ji-Kang;Kim, So-Young;Park, Kun-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1248-1252
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    • 2012
  • Potassium added with bamboo salt showed better antioxidative effects than bamboo salt, solar salt, or purified salt. It also showed inhibitory effects on the mutagenicity of MNNG (N-methyl-N-nitro-N-nitrosoguanidine) in a Salmonella Typhimurium TA100 tester strain. At concentrations of 1.25 and 2.5 mg/plate, potassium bamboo salt and bamboo salt showed weaker co-mutagenicity effects than either purified salt or solar salt, respectively. Anticancer effects of salts were evaluated using MTT assay in HCT-116 human colon carcinoma cells. At a 1% salt concentration, the growth inhibitory rate of potassium bamboo salt was 54%, higher than that of 1 time baked bamboo salt (36%). However, purified salt and solar salt showed relatively lower inhibitory effects of 19% and 23%, respectively. To determine the inhibitory mechanisms of potassium bamboo salt, the expression levels of Bax and Bcl-2 genes in HCT-116 cells were determined by RT-PCR. Potassium bamboo salt significantly increased Bax and decreased Bcl-2 expression levels unlike bamboo salt, purified salt, and solar salt (p<0.05). Therefore, addition of potassium to salt decreased co-mutagenicity and increased in vitro antioxidative and anticancer effects.

Transcriptional regulation of chicken leukocyte cell-derived chemotaxin 2 in response to toll-like receptor 3 stimulation

  • Lee, Seokhyun;Lee, Ra Ham;Kim, Sung-Jo;Lee, Hak-Kyo;Na, Chong-Sam;Song, Ki-Duk
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1942-1949
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    • 2019
  • Objective: Leukocyte cell-derived chemotaxin 2 (LECT2) is associated with several physiological processes including inflammation, tumorigenesis, and natural killer T cell generation. Chicken LECT2 (chLECT2) gene was originally identified as one of the differentially expressed genes in chicken kidney tissue, where the chickens were fed with different calcium doses. In this study, the molecular characteristics and gene expression of chLECT2 were analyzed under the stimulation of toll-like receptor 3 (TLR3) ligand to understand the involvement of chLECT2 expression in chicken metabolic disorders. Methods: Amino acid sequence of LECT2 proteins from various species including fowl, fish, and mammal were retrieved from the Ensembl database and subjected to Insilco analyses. In addition, the time- and dose-dependent expression of chLECT2 was examined in DF-1 cells which were stimulated with polyinosinic:polycytidylic acid (poly [I:C]), a TLR3 ligand. Further, to explore the transcription factors required for the transcription of chLECT2, DF-1 cells were treated with poly (I:C) in the presence or absence of the nuclear factor ${\kappa}B$ ($NF{\kappa}B$) and activated protein 1 (AP-1) inhibitors. Results: The amino acid sequence prediction of chLECT2 protein revealed that along with duck LECT2 (duLECT2), it has unique signal peptide different from other vertebrate orthologs, and only chLECT2 and duLECT2 have an additional 157 and 161 amino acids on their carboxyl terminus, respectively. Phylogenetic analysis suggested that chLECT2 is evolved from a common ancestor along with the actinopterygii hence, more closely related than to the mammals. Our quantitative polymerase chain reaction results showed that, the expression of chLECT2 was up-regulated significantly in DF-1 cells under the stimulation of poly (I:C) (p<0.05). However, in the presence of $NF{\kappa}B$ or AP-1 inhibitors, the expression of chLECT2 is suppressed suggesting that both $NF{\kappa}B$ and AP-1 transcription factors are required for the induction of chLECT2 expression. Conclusion: The present results suggest that chLECT2 gene might be a target gene of TLR3 signaling. For the future, the expression pattern or molecular mechanism of chLECT2 under stimulation of other innate immune receptors shall be studied. The protein function of chLECT2 will be more clearly understood if further investigation about the mechanism of LECT2 in TLR pathways is conducted.

Effects of Characteristics of Ovarian follicular Fluid and Ant-Inhibin Serum on Steroid Hormone Secretion by Hanwoo Granulosa Cells In Vitro (한우 난소의 Follicular Fluid의 특징과 과립막 세포의 스테로이드호르몬 분비에 대한 Anti-Inhibin Serum의 첨가효과)

  • 성환후;민관식;양병철;노환국;최선호;임기순;장유민;박성재;장원경
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.119-124
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    • 2001
  • This study was performed to investigate the effects of the peptide to carrier ratio on the immune and biological functions to inhibin immunization in Hanwoo. A peptide sequence kom the alpha -subunit (19~32 peptide) of porcine inhibin was synthesized for antigen and conjugated to human serum albumin(HSA) for carrier protein. Anti-inhibin sera(AI) were produced 52 day later from rabbit after injection of inhibin-$\alpha$ -subunit peptide conjugator for antigen with the interval of 2 weeks. Immune-blotting analysis using antibody specific fur inhibin-$\alpha$ subunits revealed that the inhibin was detected at 1.0 cm bovine follicular fluid(bFF). However, each stage of corpus lutea and 0.1 cm of follicular fluid were not detected. The maximal contents of estradiol-17 $\beta$ in Hanwoo ovarian follicular fluid were detected at 2.0 cm of follicular size(diameter), but the mean total contents of these hormone decreased significantly with decreasing diameter of follicles. However, progesterone contents of follicular fluid were high at 1.0 cm of follicle. Progesterone secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) inhibited in 5% bFF and 5% bFF + 5% AI addition group compared with control group. Estradiol-17 $\beta$ secretion by Hanwoo granulosa cell cultured for 48 hr in vitro was significantly (p<0.05) increased in 5% AI and 5% AI + 5% bFF addtion group compared with control group. However, the groups added 5% AI were not changed compared to control groups in progesterone and estradiol-17 $\beta$. Taken together, we suggested that inhibin in the mature FF plays a pivotal role on the biosynthesis of steroid hormone of follicular cells during follicular development.

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Evaluation of Embryotoxic Potential of Olaquindox and Vitamin A in Micromass Culture and in Rats

  • Kang, Hwan-Goo;Ku, Hyun-Ok;Jeong, Sang-Hee;Cho, Joon-Hyoung;Son, Seong-Wan
    • Toxicological Research
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    • v.26 no.3
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    • pp.209-216
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    • 2010
  • Limb bud (LB) and central nerve system (CNS) cells were prepared from 12.5 day old pregnant female Crj:CD (SD) rats and treated with olaquindox and vitamin A. Cytotoxicity and inhibition on differentiation were measured in each cell. Three doses of olaquindox (4, 21 and 100 mgkg), and 0.2 and 75 mg/kg of vitamin A were administered to pregnant rat for 11 days from $6^{th}$ to $16^{th}$ of pregnancy. $IC_{50}$ values of olaquindox for proliferation and differentiation in CNS cell were 22.74 and $28.32\;{\mu}g/ml$ and 79.34 and $23.29\;{\mu}g/ml$ in LB cell and those values of vitamin A were 8.13 and $5.94\;{\mu}g/ml$ in CNS cell and 0.81 and $0.05\;{\mu}g/ml$ in LB cell, respectively. Mean body weights of pregnant rats were decreased at high dose of olaquindox (110 mg/kg) but relative ovary weight, number of corpus lutea, and number of implantation were not changed. Resorption and dead fetus were increased at high dose of olaquindox, and relative ovary weight, the number of corpus lutea and implantation, and sex ratio of male to female were not significantly changed in all dose of olaquindox. Mean fetal and placenta weights were significantly (p < 0.01) decreased in rats of high group. Seven fetuses out of 103 showed external anomaly like bent tail, and 10 out of 114 fetuses showed visceral anomalies at high group. The ossification of sternebrae and metacarpals were significantly (p < 0.01) increased by low and middle dose of olaquindox but it was significantly (p < 0.01) prohibited by high dose of olaquindox. In rats treated with vitamin A, the resorption and dead fetus were increased by high dose. Mean fetal weights were significantly (p < 0.01) increased by low dose but significantly (p < 0.01) decreased by high dose. Thirty four fetuses out of 52 showed external anomaly; bent tail (1), cranioarchschisis (14), exencephaly (14), dome shaped head (22), anophthalmia (15), brcahynathia (10) and others (19). Forty five fetuses out of 52 showed soft tissue anomaly; cleft palate (42/52) and anophthalmia (22/52) by high dose of vitamin A. Sixty one fetuses out of 61 (85.2%) showed skull anomaly; defect of frontal, partial and occipital bone (21/61), defect of palatine bone (52/61) and others (50/61). In summary, we support that vitamin A is strong teratogen based on our micromass and in vivo data, and olaquindox has a weak teratogenic potential in LB cell but not in CNS cell. We provide the in vivo evidence that a high dose of olaquindox could have weak embryotoxic potential in rats.

Impact of Peri-Operative Anemia and Blood Transfusions in Patients with Gastric Cancer Receiving Gastrectomy

  • Chang, Chih-Chun;Sun, Jen-Tang;Chen, Jing-Yuan;Chen, Yi-Ting;Li, Pei-Yu;Lee, Tai-Chen;Su, Ming-Jang;Wu, Jiann-Ming;Yen, Tzung-Hai;Chu, Fang-Yeh
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1427-1431
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    • 2016
  • Background: Potential disadvantages of blood transfusion during curative gastrectomy for gastric cancer have been reported, and the role of peri-operative transfusions remains to be ascertained. Thus, the aim of our study was to survey its impact in patients with gastric cancer undergoinging gastrectomy. Materials and Methods: Clinical data of patients receiving curative gastrectomy at Far Eastern Memorial Hospital were obtained. Findings for pre-operative anemia states, pre-, peri- and post-operative transfusion of red blood cell (RBC) products as well as post-operative complication events were collected for univariate analysis. Results: A total of 116 patients with gastric cancer received gastrectomy at Far Eastern Memorial Hospital from 2011 to 2014. Both pre-operative and intra- and post-operative transfusion of RBC products were markedly associated with post-operative infectious events (OR: 3.70, 95% CI: 1.43-9.58, P=0.002; OR: 8.20, 95% CI: 3.11-22.62, P<0.001, respectively). In addition, peri- and post-operative RBC transfusion was significantly associated with prolonged hospital stay from admission to discharge (OR: 8.66, 95% CI: 1.73-83.00, P=0.002) and post-operative acute renal failure (OR: 19.69, 95% CI: 2.66-854.56, P<0.001). Also, the overall survival was seemingly decreased by peri-operative RBC transfusion in our gastric cancer cases (P=0.078). Conclusions: Our survey indicated that peri-operative RBC transfusion could increase the risk of infectious events and acute renal failure post curative gastrectomy as well as worsen the overall survival in gastric cancer cases. Hence, unnecessary blood transfusion before, during and after curative gastrectomy should be avoided in patients with gastric cancer.