• Title/Summary/Keyword: in vivo animal experiment

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In-Vitro Development of Early Stage Mouse and Bovine Embryos to Blastocysts in TCM 199 Supplemented with nonspecific Immunostimulator $Barodon-FX^{(R)}$ (비특이 면역증강제 $Barodon-FX^{(R)}$ 첨가 TCM199에서 생쥐 및 소 초기배의 체외 배반포 발달에 관한 연구)

  • 정영채;나광빈;김창근;류재원;최수일;전경수;류범룡
    • Korean Journal of Animal Reproduction
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    • v.25 no.2
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    • pp.131-138
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    • 2001
  • This experiment was designed to evaluate effects of nonspecific immunostimulator(NIS) Barodon-FX(equation omitted), anionic alkali mineral complex and far-infrared radiation solution on in vivo-produced mouse and in vitro-produced bovine embryos to blastocyst development. Proportion of mouse embryos developing into blastocyst was not greater in BSA- and Barodon-added medium than in BSA-control, but there was signifcantly different(P < 0.05) in hatching and hatched blastocyst development between 0.25% Barodon-and PVP-contained medium(54.7%) than PVP-control(32.5%). BOEC and GC resulted in higher proliferation rate(24∼40% and 17∼22%, respectively) in 0.25∼0.5% Barodon-added medium than in controls, but proliferation of GC and CC greatly decreased in 1∼2% Barodon-added medium. Effect of Barodon on cell proliferation greatly varied among somatic cells. Proportion of early bovine embryos developing into morula and blastocyst was significantly greater(P < 0.05) in 0.5% Barodon-added medium(50% and 63.6%) than in control(31.6% and 27.4%) under co-culture with BOEC and GC, but developmental rate was not different between other Barodon treatments and control. These data indicate that effect of Barodon on cell proliferation significantly varied between somatic cells and that addition of 0.5% Barodon in BOEC-coculture system may further improve blastocyst development in early bovine embryos.

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Effects of β-Glucan from Paenibacillus polymyxa and L-theanine on Growth Performance and Immunomodulation in Weanling Piglets

  • Hwang, Y.H.;Park, B.K.;Lim, J.H.;Kim, M.S.;Song, I.B.;Park, S.C.;Jung, H.K.;Hong, J.H.;Yun, H.I.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.12
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    • pp.1753-1759
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    • 2008
  • Forty weanling piglets ($5.6{\pm}0.5kg$ and 26 to 30 d of age) were used in a 28-d experiment to determine the effects of ${\beta}$-glucan from Paenibacillus polymyxa and L-theanine on growth performance. Piglets were randomly allotted to four groups (n = 10, 2 animals per pen) provided with the basal feed (control), ${\beta}$-glucan 400 mg/kg feed, L-theanine 80 mg/kg feed or ${\beta}$-glucan plus l-theanine (combination of the above-mentioned concentrations). Body weight and feed consumption were recorded during four weeks. Subsequently, the immunomodulatory effects of ${\beta}$-glucan and L-theanine were investigated for lipopolysaccharide (LPS)-induced cytokine production in vitro and in vivo on day 28. Although there were no significant differences in the growth performances among the treatment groups, ${\beta}$-glucan plus L-theanine had 5.6% greater ADG (p = 0.074) on day 21 to 28. ${\beta}$-Glucan alone or plus L-theanine increased interleukin (IL)-10 levels and decreased interferon (IFN)-$\gamma$ and tumor necrosis factor (TNF)-${\alpha}$ levels in cultured medium by LPS treatment (p<0.05). Plasma IL-10 levels were also increased in the piglets fed with ${\beta}$-glucan alone or plus L-theanine after LPS challenge ($25{\mu}g/kg$, i.p.), whereas plasma IFN-$\gamma$ and TNF-${\alpha}$ levels were decreased (p<0.05). The levels of IFN$\gamma$ in piglets fed with ${\beta}$-glucan plus L-theanine showed the greatest inhibition after LPS challenges. In conclusion, treatment of ${\beta}$-glucan alone or plus L-theanine might lessen inflammatory responses against Gram-negative bacterial infection via the inhibition of pro-inflammatory cytokine production and enhancement of anti-inflammatory cytokine production. Further studies are needed to determine an optimal concentration of ${\beta}$-glucan and L-theanine for improved growth performance.

Normal Pregnancy of Mouse Embryos Transferred after Assisted Hatching by a 1.48$\mu\textrm{m}$ Diode Laser (1.48$\mu\textrm{m}$ Diode Laser로 보조 부화처리 후 이식된 생쥐배의 정상임신에 관한 연구)

  • 김은영;이봉경;남화경;이금실;윤산현;박세필;정길생;임진호
    • Korean Journal of Animal Reproduction
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    • v.22 no.3
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    • pp.287-292
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    • 1998
  • The objective of this study was to test whether ZP drilling using a 1.48$\mu$m diode laser beam on mouse IVF embryos becomes effective the hatching and normal in vivo development, as a preliminary test for obtaining the additional proof that the 1.48$\mu$m diode laser could be used safely for human applications. The results obtained in this experiment were as follows: when the hatched rates of mouse embryos by laser ZP drilling according to the embryonic stage were examined until 72 hr (in case of blast tocyst: day 4 after IVF) or 120 hr (in case of 4-cell: day 2 after IVF) after treatment, the d data of laser drilled blastocysts (81.8%) was significantly higher than those of control (hatching blastocyst: day 4 after IVF) (54.2%) and laser drilled 4-cell embryos (45.5%) (p<0.05). When the effect of laser drilling on implantation rates following embryo transfer in day 3 synchronized pseudopregnant recipients was examined, the l laser drilled group (48.7%) was slightly higher than that of control group (43.6%). In addition, when the several pregnant mice delivered in two groups were analysed their chromosomal normality and tested reproductive ability, all p pups were presented normal chromosomal number (n=40) and showed normal growth and reproductive ability. Therefore, these results dem-onstrated that ZP drilling using a 1.48$\mu$m diode l laser can increase the embryo hatching and ind duce the normal pregnancy of mouse embryos.

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In Vitro Development of Somatic Cell Nuclear Transfer Embryo Treated with Flavonoid and Production of Cloned Jeju Black Cattle (플라보노이드 처리된 체세포 핵이식 배아의 체외 발달 및 제주흑우 복제 소 생산)

  • Kim, Eun-Young;Kim, Yeon-Ok;Kim, Jae-Youn;Park, Min-Jee;Park, Hyo-Young;Han, Young-Joon;Mun, Seong-Ho;Oh, Chang-Eon;Kim, Young-Hoon;Lee, Sung-Soo;Ko, Moon-Suck;Park, Se-Pill
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.127-134
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    • 2010
  • This study was to investigate the effect of flavonoid treatment on in vitro development of bovine somatic cell nuclear transfer (SCNT) embryos, and their pregnancy and delivery rate after embryo transfer into recipient. In experiment 1, to optimize the flavonoid concentration, parthenogenetic day 2 ($\geq$ 2-cell) embryos were cultured in 0 (control), 1, 10 and $20\;{\mu}M$ flavonoid for 6 days. In the results, in vitro development rate was the highest in $10\;{\mu}M$ flavonoid group (57.1%) among treatment groups (control, 49.5%; $1\;{\mu}M$, 54.2%; $20\;{\mu}M$, 37.5%), and numbers of total and ICM cells were significantly (p<0.05) higher in $10\;{\mu}M$ flavonoid group than other groups. We found that $10\;{\mu}M$ flavonoid treatment can significantly (p<0.05) decrease the apoptotic index and derive high expression of anti-oxidant, anti-apoptotic, cell growth and development marker genes such as Mn-SOD, Survivin, Bax inhibitor, Glut-5, In-tau, compared to control group. In experiment 2, to produce the cloned Jeju Black Cattle, beef quality index grade 1 bull somatic cells were transferred into enucleated bovine MII oocytes and reconstructed embryos were cultured in $10\;{\mu}M$ flavonoid added medium. When the in vitro produced day 7 or 8 SCNT blastocysts were transferred into a number of recipients, $10\;{\mu}M$ flavonoid treatment group presented higher pregnancy rate (10.2%, 6/59) than control group (5.9%, 2/34). Total three cloned Jeju Black calves were born. Also, two cloned calves in $10\;{\mu}M$ flavonoid group were born and both were all healthy at present, while the one cloned calf born in control group was dead one month after birth. In addition, when the result of short tandem repeat marker analysis of each cloned calf was investigated, microsatellite loci of 11 numbers matched genotype between donor cell and cloned calf tissue. These results demonstrated that the flavonoid addition in culture medium may have beneficial effects on in vitro and in vivo developmental capacity of SCNT embryos and pregnancy rate.

The Inhibitory Effects of $Chrthami$ Semen Oil Pharmacopuncture (CSOP) on Synovial Membranes in Type II Collagen-Induced Arthritis Mice (홍화자유약침이 CIA 모델 생쥐의 윤활관절막 손상 억제에 미치는 영향)

  • Back, Seong-Wook;Kim, Eun-Jung;Hwang, Ji-Hoo;Yoon, Jong-Hwa;Lee, Seung-Deok;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.29 no.1
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    • pp.115-125
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    • 2012
  • Objectives : The purpose of this study is to observe the inhibitory effects of $Chrthami$ semen oil pharmacopuncture(CSOP) on CIA (collagen-induced arthritis) mice. Materials and Methods : Two types of experiments were conducted: $in$ $vitro$ assay, inhibition of MIF mRNA and TNF-${\alpha}$ mRNA expressions in synovial membranes was observed, and $in$ $vivo$ assay, $1{\mu}{\ell}/kg$ CSOP was injected every day to the left $Weizhong$ ($BL_{40}$) from day 3 to 21 after induction of CIA, and changes in paw edema, apical surface morphology, neovascularization in synovial membranes, fibrosis, pro-inflammatory cytokines production, Th-1 differentiation, and anti-inflammatory effect were investigated. Results : 1. In synoviocytes of the CIA mice treated with CSOP, MIF mRNA and TNF-${\alpha}$ mRNA expressions were down-regulated in a dose-dependent manner. 2. Paw edema of the CIA mice treated with CSOP was diminished. 3. Tissue injury in the synovial membranes, capillary distribution and fibrosis were reduced in CSOP-treated mice. 4. MIF, TNF-${\alpha}$, IL-6, MMP-9 expressions were repressed in CSOP-treated mice during the experiment to observe the inhibitory effect on cytokines production in early stage RA. 5. IL-12 and CD28 were reduced in CSOP-treated mice during the observation of inhibitory effect on Th 1 differentiation. 6. PPAR-${\gamma}$ was increased during the experiment to observe the anti-inflammatory effect of CSOP. Conclusions : The results may suggest that administration treatment using $Chrthami$ semen oil pharmacopuncture decreases the inflammatory response on an Animal Model with CIA.

Protective Effects of Plant Extracts on the Hepatocytes of Rat Treated with Carbon Tetrachloride (사염화탄소를 투여한 흰쥐의 간세포에 대한 식물 추출물들의 보호효과)

  • Ham, Young-Kook;Kim, Sung-Wan
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.8
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    • pp.1246-1251
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    • 2004
  • To investigate the effects of plant extracts on the protection against liver damage by $CCl_4$ in rat, two kinds of experiment were performed, firstly by the primary hepatocyte culture and secondly by the animal feeding. The primary hepatocyte culture with the extracts of pine leaf, soybean sprout and mugwort showed significantly low activities (p<0.01∼0.05) of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), indicating an excellent protective effect against liver damage by $CCl_4$. In the second experiment, the microsomal malondialdehyde (MDA) contents of the above same groups were also significantly lower (p<0.01) than the $CCl_4$-treated group without plant extracts, but shiitake showed less effect. Among four kinds of plant extracts, extracts of pine leaf and mugwort showed also much higher activities of the microsomal cytochrome P-450 in comparison to soybean sprout and shiitake. In the test of xanthine oxidase (XOD) activity, all of three groups except shiitake showed significantly low activities (p<0.01). These consistent results in vitro and in vivo suggest that the extracts of pine leaf, soybean sprout and mugwort may have strong protective effects against liver damage induced by the potential toxicants such as $CCl_4$.

Effect of Haepyoijin-tang on Airway Mucin Secretion, Production, Gene Expression and Hypersecretion of Mucus (해표이진탕이 기도 뮤신의 분비, 생성 및 유전자 발현에 미치는 영향)

  • Suk, Yun Hee;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.29 no.3
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    • pp.65-79
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    • 2015
  • Objectives : In this study, effects of haepyoijintang (HIJ) on the increase in airway epithelial mucosubstances of rats and ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells were investigated. Methods : Hypersecretion of airway mucus was induced by exposure of rats to $SO_2$ during 3 weeks. Effect of orally-administered HIJ during 2 weeks on increase in airway epithelial mucosubstances from tracheal goblet cells of rats was evaluated using histopathological analysis after staining the epithelial tissue with PAS-alcian blue. Possible cytotoxicity of HIJ was evaluated by examining the potential damage of kidney and liver functions by measuring serum GOT/GPT activities and serum BUN and creatinine concentrations of rats and the body weight gain during experiment, after administering HIJ orally. At the same time, the effect of HIJ on ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production and gene expression from human airway epithelial cells (NCI-H292) were investigated. Confluent NCI-H292 cells were pretreated for 30 min in the presence of HIJ and treated with ATP ($200{\mu}M$), PMA (10 ng/ml), EGF (25 ng/ml) or TNF-${\alpha}$ (0.2 nM) for 24 hrs, to evaluate the effect of HIJ both on ATP-, PMA-, EGF- or TNF-${\alpha}$-induced MUC5AC mucin production using enzyme-linked immunosorbent assay (ELISA) and on gene expression by the same inducers using reverse transcription-polymerase chain reaction (RT-PCR). Results : (1) HIJ decreased the amount of intraepithelial mucosubstances of trachea of rats. (2) HIJ did not show renal and hepatic toxicities and did not affect body weight gain of rats during experiment. (3) HIJ significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin productions from NCI-H292 cells. (4) HIJ significantly inhibited ATP-, PMA-, EGF-, and TNF-${\alpha}$-induced MUC5AC mucin gene expression from NCI-H292 cells. Conclusions : The result from the present study suggests that HIJ might control the production and gene expression of airway mucin observed in various respiratory diseases accompanied by mucus hypersecretion and do not show in vivo toxicity to liver and kidney functions after oral administration. Effect of HIJ with their diverse components should be further investigated using animal experimental models that can reflect the pathophysiology of airway diseases through future studies.

Protective Effects of Extracts of Protaetia brevitarsis on Carbon tetrachloride-induced Hepatotoxicity in the Mice (사염화탄소에 의해 유발된 흰쥐의 간독성에 미치는 흰점박이꽃무지 추출물의 간보호 효과)

  • Chon, Jeong-Woo;Kweon, HaeYong;Jo, You-Young;Yeo, Joo Hong;Lee, Heui Sam
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.93-100
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    • 2012
  • To investigate the effects of Protaetia. brevitarsis extracts on the protection against liver damage by carbon tetrachloride($CCl_4$) in rat, two kinds of experiment were performed, firstly by the primary hepatocyte culture and secondly by the animal feeding. The primary hepatocyte culture with the extracts of P.brevitarsis showed significantly low activities of GPT, bile acid, and bilirubin, indicating an excellent protective effect against liver damage by $CCl_4$. Especially, below molecular weight 1,000 blew the water to have 32.1% recovery degree. In the seconde experiment, serum GPT activity was significantly decreased in water fraction of P. brevitarsis compared to $CCl_4$ treatment by 98.2%. Serum concentration of bile acid and bilirubin were tended to increased by $CCl_4$ treatment, but water fraction of P. brevitarsis and silymarin recovered the level. These consistent results in vitro and in vivo suggest that the extracts of P. brevitarsis may have strong protective effects against liver damage induced by the potential toxicants such as $CCl_4$.

Effect of Riboflavin Tetrabutylate on the Activity of Drug Metabolizing Enzyme and Lipid Peroxidation in Liver Microsomes of Rats (Riboflavin Tetrabutylate가 약물대사 효소 및 지질 과산화효소에 미치는 영향)

  • Lee, H.W.;Kim, W.J.;Hong, S.S.;Kwack, C.Y.;Hong, S.U.
    • The Korean Journal of Pharmacology
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    • v.16 no.2 s.27
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    • pp.45-53
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    • 1980
  • Lipid peroxidation in vitro has been identified as a basic deteriorative reaction in cellular mechanism of aging processes, such as air pollution oxidant damage to cell and to the lung, chlorinated hydrocarbon hepatotoxicity. Many experimental evidences were reported by several investigators that lipid peroxidation could be one of the principle causes for the hepatotoxicity produced by $CCl_4$. It is now reasonably established that $CCl_4$ is activated to a free radical in vivo, that lipid peroxidation occurs very quickly in microsomes prepared from damaged livers, that the peroxidation is associated with loss of enzyme activity of microsomes, and that various antioxidants can protect animals against the hepatotoxic effect of $CCl_4$. Recent studies have drawn attention to some other feature of microsomal lipid peroxidation. Incubation of liver microsomes in the presence of NADPH has led to a loss of cytochrome $P_{450}$. However, the presence of an antioxidant prevented lipid peroxidation and preserved cytochrome $P_{450}$. Decrease of cytochrome $P_{450}$ in microsomes under in vitro incubation can be enhanced by $CCl_4 and these changes were parallel to a loss of microsomal polyunsaturated fatty acid and formation of malonaldehyde. The primary purpose of this experiment was to study the effect of riboflavin tetrabutylate on lipid peroxidation, specially, the relationship between lipid peroxidation and drug metabolizing enzyme system which is located in smooth endoplasmic recticulum as well as the effect of ritoflavin tetrabutylate on drug metabolizing enzyme system of animal treated with $CCl_4$. Albino rats were used for experimental animal. In order to induce drug metabolizing enzyme system, phenobarbital was injected intraperitoneally. $CCl_$ and riboflavin tetrabutylate were given intraperitoneally as solution in olive oil. Microsomal fraction was isolated from liver of animals and TBA value as well as the activity of drug metabolizing enzyme were measured in the microsomal fractions. The results are summerized as following. 1) The secobarbital induced sleeping time of $CCl_4$ treated rat was about 2 times longer than that of the control group. However, the pretreatment with riboflavin tetrabutylate inhibited completely the lengthened sleeping time due to $CCl_4$ treatment. Furthermore TBA value was significantly increased in $CCl_4$ treated rat in comparison to control group tut the increase of TBA value was prevented by the pretreatment with riboflavin tetrabutylate. On the other hand, the activity of hepatic drug metabolizing enzyme was decreased in $CCl_4$ group, however, the pretreatment with riboflavin tetrabutylate also prevented the decrease of the enzyme activity caused by $CCl_4$. 2) The effect of riboflavin tetrabutylate on TBA value and the activity of drug metabolizing enzyme in vitro was similar to in vivo results. Incubation of liver microsome from rat in the presence of $CCl_4$, $Fe^{++}$, or ascorbic acid has led to the marked increase of TBA value, however, the addition of riboflavin tetrabutylate in incubation mixture prevented significantly the increase of TBA value, suggesting the inhibition of lipid peroxidation. In accordance with TBA value, the activity of drug metabolizing enzyme was inhibited in the presence of $CCl_4$, $Fe^{++}$, ascorbic acid but the addition of riboflavin tetrabutylate protected the loss of the enzyme activity in microsome under in vitro incubation.

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Comparison of TME Values of Imported Corns and ME Values Determined by In vitro Method (수입 옥수수들의 TME가와 In vitro 방법에 의해 측정한 ME가의 비교)

  • Cho, J.H.;Paik, I.K.;Hyun, Y.
    • Journal of Animal Science and Technology
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    • v.49 no.6
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    • pp.737-744
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    • 2007
  • An experiment was conducted to compare the ME values of imported corns measured by In vivo(TME) and In vitro(MEn, ME and MEpc) methods and to investigate the true amino acid availability(TAAA) and the true nutrient availability(TNA) of imported corns. For TME assay, twenty four 57-weeks-old Hy-Line roosters were assigned to fasting group(4 roosters) and four corn groups(5 roosters each): USA; corn produced in the United States, ARG; corn produced in Argentina, CHN; regular corn produced in China, CHNP; premium corn produced in China. The MEn, ME and MEpc values were determined by equations based on chemical analysis. The TME value of USA(3,745kcal/kg) in as fed basis was significantly (P<0.01) higher than ARG(3,555kcal/kg) and CHNP(3,518kcal/kg) but was not significantly different from CHN(3,671kcal/kg). The TME value of USA(4,144kcal/kg) in DM basis was not significantly different from CHN(4,060kcal/kg) and CHNP(4,008kcal/kg) but was significantly(P<0.05) higher than ARG(4,001kcal/kg). There were significant differences in TAAA of phenylalanine, histidine and arginine among imported corns. Those of USA were highest but overall TAAA was not significantly different among imported corns. True availability of NFE of USA, ARG and CHN was significantly(P<0.05) higher than that of CHNP. However, true availability of crude protein, crude fat, crude fiber and crude ash were not significantly different among corns. The correlation coefficient between TME and MEn value was 0.91 which was significant at P<0.1 but correlation coefficient between TME and ME value and between TME and MEpc value was 0.90 and 0.83, respectively which was not significant at P<0.1. In conclusion, US corn was highest in TME values and Chinese premium corn was not significantly different from regular Chinese corn. The MEn value obtained by equation based on chemical analysis may be used as a tool to evaluate TME value of corn.