• Title/Summary/Keyword: ANIMAL MODEL

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Aberrant Methylation of p16 Tumor Suppressor Gene and Death-Associated Protein Kinase in Non-Small Cell Lung Carcinoma (비소세포폐암 조직에서 p16 종양억제유전자와 Death-Associated Protein Kinase의 Aberrant Methylation의 양상)

  • Kim, Yun-Seong;Lee, Min-Ki;Jung, Kyung-Sik;Kim, Ki-Uk;Kim, Young-Dae;Lee, Hyung-Ryul;Lee, Chang-Hoon;Seok, Ju-Won;Kim, Yong-Ki;Jun, Eun-Sook;Choi, Young-Min;Rha, Seo-Hee;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.51 no.2
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    • pp.108-121
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    • 2001
  • Background : The $p16^{INK4a}$ (p16) twnor suppressor gene is frequently inactivated in hwnan non-small cell lung cancers (NSCLCs), predominantly through homozygous deletion or in association with aberrant promotor hypermethylation. Death-associated protein kinase (DAPK) gene influences interferon $\gamma$-induced apoptotic cell death and has important role in metastasis of lung cancer in animal model. Hypermethylation of promoter region of DAP kinase gene may suppress the expression of this gene. Methods : This study was performed to investigate the aberrant methylation of p16 or DAP kinase in 35 resected primary NSCLCs by methylation-specific PCR (MSP), and demonstrated frequency, diagnostic value and clinical implication of aberrant methylation of two genes. Results : Thirty-two cases were male patients, and 3 cases were female patients with an average age was 57. $8{\pm}10.5$ years. The histologic types of lung cancer were 22 of squamous cell carcinoma, 12 of adenocarcinoma, 1 of large cell carcinoma. Pathologic stages were 11 cases of stage I (1 IA, 10 IB), 13 cases of stage II (1 IIA, 12 IIB), and 11 cases of stage III (9 IIIA, 2 IIIB). Regarding for the cancer tissue, p16 aberrant methylation was noted in 13 case of 33 cases (39.4%), DAP kinase in 21 cases of 35 cases (60%). Age over 55 year was associated with p16 aberrant methylation significantly (p<0.05). Methylation status of two genes was not different by smoking history, histologic type, size of tumor, lymph node metastasis and disease progression of lung cancer. There was no correlation between p16 and DAP kinase hypermethylation. Conclusion: This investigation demonstrates that aberrant methylation of p16 tumor suppressor gene or DAP kinase showed relatively high frequency (74.3%) in NSCLCs, and that these genes could be a biologic marker for early detection of lung cancer.

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Changes of Serum Fatty Acid and Carnitine Levels after Administration of L-carnitine in Rats (흰쥐에서 L-carnitine 투여 후에 혈청 지방산과 Carnitine의 농도 변화)

  • Lee, Jae Won;Hong, Young Mi
    • Clinical and Experimental Pediatrics
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    • v.45 no.9
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    • pp.1075-1082
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    • 2002
  • Purpose : Obesity is known to be associated with hypertension, dyslipidemia, and fatty liver and is thought to be associated with increased levels of free fatty acids. One of the strategies for decreasing free fatty acid levels is stimulation of hepatic lipid oxidation with L-carnitine. Carnitine is an essential cofactor for transport of long-chain fatty acid into mitochondria for oxidation. This study was designed to evaluate the changes of serum fatty acids and carnitine levels after exogenous injection of L-carnitine. Methods : Sprague Dawley rats were divided into two groups. Group A was control. Group B was given intraperitoneal injection with L-carnitine(200 mg/kg) daily for two weeks. Serum lipid (total cholesterol, triglyceride, HDL-cholesterol, LDL-cholesterol) and fatty acid levels were analyzed on the first day of the first and second weeks after injection of L-carnitine. Total, free, and acyl carnitine levels also were performed by a enzymatic cycling techniques at the same day intervals. Results : There was no significant difference between the two groups in total cholesterol, HDL-cholesterol, LDL-cholesterol levels before and after the administration of L-carnitine. But triglyceride levels were significantly decreased at the first week in group B compared with group A. Among free fatty acids, linoleic acid showed significant decrement(A group : $131.3{\pm}31.3mg/dL$ vs B group : $90.0{\pm}7.0mg/dL$) at the first week. Total, free, and acyl carnitine levels showed significant increments at all days intervals, but only free carnitine showed significant increments according to cumulative doses of carnitine. Conclusion : Plasma linoleic acid, a long-chain fatty acid, showed significant decrement after administration of L-carnitine in the first week. This may suggest that L-carnitine can be used as an antilipidemic agent for obese patients. A prospective study will investigate obese children in the future.

Effect of Onion Peel Extracts on Blood Lipid Profile and Blood Coagulation in High Fat Fed SD Rats (고지방 섭취 흰쥐에서 양파 껍질 추출물의 보충 섭취가 혈중 지질농도와 혈행 개선에 미치는 효과)

  • Chung, Hye-Kyung;Shin, Min-Jeong;Cha, Yong-Jun;Lee, Kyung-Hea
    • The Korean Journal of Food And Nutrition
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    • v.24 no.3
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    • pp.442-450
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    • 2011
  • Numerous studies have suggested that dietary flavonoids contribute to prevent cardiovascular disease. Onion contains many functional phytochemicals such as quercetin. The aim of this study was to examine whether onion peel extracts supplementation affect blood lipid profiles and blood coagulation in animal model. Total 48 Sprague-Dawley male rats at 5 weeks old were divided into 6 groups with different diets(C: control, HF: high fat diet, HFOE 0.01%: high fat+onion peel extract 0.01% diet, HFOE 0.02%, HFOE 0.05%, HFOE 0.1%) for 8 weeks. Onion peel extract supplementation significantly decreased serum levels of LDL-cholesterol and increased HDL-cholesterol, while total cholesterol and triglyceride levels were not affected. Hematological parameters(hematocrit, white blood cell, red blood cell, and platelet count) and blood coagulation parameters(prothrombin time, activated partial thromboplastin time, thrombin time, and fibrinogen) were not significantly different among 6 groups. However, activated partial thromboplastin time of HFOE 0.05% group was significantly longer than that of HF group. These results indicate that onion peel extract supplementation displays hypocholestrolemic effects but does not seem to have anti-coagulation effects in high fat fed SD rats.

Antioxidative Activities of the Codonopsis lanceolata Extract in vitro and in vivo (더덕(Codonopsis lanceolata) 추출물의 in vitro 및 in vivo 항산화 효과)

  • Kim, Soo-Hyun;Chung, Mi-Ja;Jang, Hae-Dong;Ham, Seung-Shi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.2
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    • pp.193-202
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    • 2010
  • In vitro activities of Codonopsis lanceolata (CL) 70% ethanol extract and its fractions (hexane, chloroform, ethyl acetate, butanol and water) were examined by total polyphenol content, reducing power, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) (ABTS), 2,2-diphenyl-$\beta$-picrylhydrazyl (DPPH), and oxygen radical absorbance capacity (ORAC) assays. The ethyl acetate fraction from CL ethanol extract (CLEA) showed the highest total polyphenol content (22.7 mg/g) among five fractions, and also exhibited an excellent reducing power (0.42~1.27 at $250\sim1,000\;{\mu}g/mL$). CLEA at $100\sim400\;{\mu}g/mL$ concentrations had 27.7~70.3% of ABTS radical scavenging activity and the highest DPPH radical scavenging activity (81.6% at $400\;{\mu}g/mL$). CLEA had dominantly higher $ORAC_{{ROO}{\cdot}}$activity compared to other fractions. CLEA and butanol fraction had significantly higher $ORAC_{{OH}{\cdot}}$ activities than 70% ethanol extract, hexane, chloroform and water fractions. The CLEA exhibited the highest antioxidant activity in CL 70% ethanol extract and its fractions. Thus, effect of CLEA treatment on antioxidant gene expression under the oxidative stress conditions by a high fat diet in animal model was studied by microarray and RT-PCR methods. The 31 antioxidant genes were expressed but the genes were not up-regulated at least a two-fold by CLEA treatment. We concluded that CLEA does not have an indirect antioxidant effect but a direct antioxidant effect by up-regulation of antioxidant genes in high fat diet-induced obese mice.

Virus Inactivation during the Manufacture of a Collagen Type I from Bovine Hides (소 가죽 유래 Type I Collagen 생산 공정에서 바이러스 불활화)

  • Bae, Jung Eun;Kim, Chan Kyung;Kim, Sungpo;Yang, Eun Kyung;Kim, In Seop
    • Korean Journal of Microbiology
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    • v.48 no.4
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    • pp.314-318
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    • 2012
  • Most types of collagen used for biomedical applications, such as cell therapy and tissue engineering, are derived from animal tissues. Therefore, special precautions must be taken during the production of these proteins in order to assure against the possibility of the products transmitting infectious diseases to the recipients. The ability to remove and/or inactivate known and potential viral contaminants during the manufacturing process is an ever-increasingly important parameter in assessing the safety of biomedical products. The purpose of this study was to evaluate the efficacies of the 70% ethanol treatment and pepsin treatment at pH 2.0 for the inactivation of bovine viruses during the manufacture of collagen type I from bovine hides. A variety of experimental model viruses for bovine viruses including bovine herpes virus (BHV), bovine viral diarrhea virus (BVDV), bovine parainfluenza 3 virus (BPIV-3), and bovine parvovirus (BPV), were chosen for the evaluation of viral inactivation efficacy. BHV, BVDV, BPIV-3, and BPV were effectively inactivated to undetectable levels within 1 h of 70% ethanol treatment for 24 h, with log reduction factors of ${\geq}5.58$, ${\geq}5.32$, ${\geq}5.11$, and ${\geq}3.42$, respectively. BHV, BVDV, BPIV-3, and BPV were also effectively inactivated to undetectable levels within 5 days of pepsin treatment for 14 days, with the log reduction factors of ${\geq}7.08$, ${\geq}6.60$, ${\geq}5.60$, and ${\geq}3.59$, respectively. The cumulative virus reduction factors of BHV, BVDV, BPIV-3, and BPV were ${\geq}12.66$, ${\geq}11.92$, ${\geq}10.71$, and ${\geq}7.01$. These results indicate that the production process for collagen type I from bovine hides has a sufficient virus-reducing capacity to achieve a high margin of virus safety.

Dietary Effect of Silk Protein on Ceramide Synthesis and the Expression of Ceramide Metabolic Enzymes in the Epidermis of NC/Nga Mice (실크단백질의 식이 공급이 아토피 피부염 동물 모델 NC/Nga Mice 피부의 세라마이드 함량 및 관련인자 발현에 미치는 영향)

  • Park, Kyung-Ho;Choi, Young-Sim;Kim, Hyun-Ae;Lee, Kwang-Gill;Yeo, Joo-Hong;Jung, Do-Hyun;Kim, Sung-Han;Cho, Yun-Hi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.5
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    • pp.554-562
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    • 2007
  • Ceramide rich intercellular lipid lamellae are thought to be particularly important in maintaining the structural integrity of epidermal barrier. Ceramide is synthesized de novo by serine palmitoyltransferase (SPT) phospholipid intermediates, serine and palmitic acid persist within the stratum corneum. The ceramide which is synthesized is degraded with fatty acid and sphingosine by degradative enzyme ceramidase. The depletion of ceramide in stratum corneum was reported in the atopic dermatitis. As an effort to search for the dietary source for improving the level of ceramide in epidermis, the dietary effects of various-typed silk protein were compared. Seventy male NC/Nga mice, an animal model of atopic dermatitis, were divided into seven groups: group CA as an atopic control with control diet, group S: 1% crude sericin diet, group F: 1% crude fibroin diet, group PS : peptide pattern of sericin(Mw 5000), group PF: peptide pattern of fibroin (Mw 1500), group AS: manufactured the same as amino acid profile of sericin and group AF: manufactured the same as amino acid profile of fibroin. Ten male BALB/c mice were served as group C (control group) control diet. All mice were fed on diet and water ad libitum for 10 weeks. Dry skin condition was established in group CA as ceramide content was decreased. Despite a marked decrease of mRNA and prorein expression of SPT, enzyme do novo synthesis, ceramide content of group S was dramatically increased by inhibiting the mRNA and protein expression of degradative enzyme ceramidase. However, dietary supplementation of crude silk fibroin protein (group F) and in other groups that were supplemented with either amino acid or peptide type of sericin or fibroin did not increase the level of ceramide. Together, our data demonstrate that dietary supplementation of crude sericin is more effective at improving ceramide level in epidemis of NC/Nga mice.

Effect of X-Irradiation on the Levels of some Sulfhydryl Groups, Protein and Cell Volume of Ehrlich Ascites Tumour Cells (X-선(線) 조사(照射)가 Ehrlich 암세포(癌細胞)의 용적(容積), 단백양(蛋白量) 및 수종(數種) Sulfhydryl 기(基)에 미치는 영향(影響)에 관(關)하여)

  • Yu, Choon-Shik;Choo, Young-Eun
    • The Korean Journal of Physiology
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    • v.3 no.2
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    • pp.9-16
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    • 1969
  • It is well known that a number of -SH and -SS containing substances afford a certain measure of protection against radiation effects in many biological systems, and it is conceivable that inherent -SH levels in Ehrlich ascites tumour (ELD)cells may be of decisive improtance with respect to the development of cellular radiation injury. So far, little effort has been directed to elucidate the changes in levels of different -SH and -SS groups in ELD cells when the tumour-bearing whole animal was subjected to the sublethal dose of X-irradiation. The present study was designed to bring some lights in the possible changes of and relationship between various sulfhydryl levels, such as P-SH, NP-SH and NP-SS, as well as the content of protein and cell volume of ELD cells, after subjecting the ELD mice to 1,200 r of X-irradiation. The animals used in this experiment were all mixed bred mice of $20{\sim}25\;gm$ in body weight (approximately 2 months old) irrespective of sex. 12 mice in one experiment were inoculated intraperitoneally with 0.2 ml of ascites tumour cells $(2{\times}10^6\;cells)$, and on the 7th day of the tumour growth, they were X-irradiated with 1,200 r, using the conventional X-ray machine under the following conditions: 200 Kv at 15 mA, 0.5 mm Cu filter, target-skin distance: 50 cm. Radiation dose was measured with the the Philip integrating dosimeter. At 24, 36, 48 and 60 hours after the X-irradiation, the mice were killed by cervical dislocation, and the tumours were taken out. Freshly withdrawn ascites tumours were placed in ice, and immediately the cell concentration was measured with the Coulter Cell Counter (Model B), and the hematocrit of the tumour cells were also determined. Cell volume was thus calculated by the cell concentration and hematocrit value. P-SH content of ELD cells was measured potentiometrically according to the method of Calcutt & Doxey, and NP-SH and NP-SS contents were measured spectrophotometrically by the method described by Ellman. Protein content of ELD cells was determined with the Folin phenol reagent by Lowry et al. Altogether, 48 experimental mice were used, and 12 mice with the only exception of X-irradiation were used as the control. Results obtained indicate that the contents of all the cellular sulfhydryl groups as well as cell volume and protein content of the ELD cells increase significantly as time progresses after the sub-lethal X-ray dose of 1,200 r was given and that all the increase is in a lineal fashion. The regression lines of the relative values, (i. e., taking each control value as 1) of all the values obtained, and the regression lines of cell volume, protein and NP-SH are identical, whereas those of NP-SS and P-SH appear to be widely seperated. However, the difference of those two lines (NP-SS & P-SH) were found to be not significant statistically (p>0.05). Therefore, it can be concluded from the above results that all the values examined increase in a lineal fashion with no statistically significant difference among them. Also, with the radiation dose of 1,200 r, the ELD cell becomes enlarged and swollen progressively up to 60 hours post-irradiation and it becomes more than two times of the original normal size at 60 hours after the irradiation, and up to this stage, it seems apparent that the cell division has been slow due to the X-irradiation applied in this experiment. It is well understandable that the contents of NP-SH, NP-SS, P-SH and protein of the ELD cells increase in parallel with the increase of the cell volume by the X-ray does used, but it also seems interesting to note that all the cellular substances tested show no appreciable difference in the pattern of increase.

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Effect of Brown Algae (Undaria pinnatifida)-Noodle on Lipid Metabolism in Serum of SD-Rats (미역 (Undaria pinnatifida) 국수가 SD계 흰쥐의 혈청 지질대사에 미치는 영향)

  • CHOI Jin-Ho;KIM Dong-Woo;KIM Jung-Hwa;KIM Dae-Ik;KIM Chang-Mok
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.1
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    • pp.42-45
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    • 1999
  • Our previous studies have demonstrated the biological effects of alginic acid as a brown algae (Undaria pinnatifida) component on inhibitory action of obesity using animal model. Sprague-Dawely (SD) male rats were fed experimental diets ($10\%,\;20\%$ and $40\%$ brown algae noodle: BA-noodles) for 4 weeks. Effects of BA-noodles on triglyceride (TG), total cholesterol. LDL and HDL-cholesterol, and atherogenic ($10\%\~15\%$ and $15\%\~18%$, respectively) by administration of $10\%,\;20\%$and $40\%$ BA-noodles compared with control group. HDL-cholesterol contents in serum of SD rats for 4 weeks significantly increased in ranges of 7 to $16\%$ by administration of $10\%,\;20\%$ and $40\%$ BA-noodles compared with control group. Atherogenic index (AI) in serum of SD rats as a initial attack markers of chronic degenerative disease for 4 weeks remarkably decreased in ranges of 14 to $28\%$ by the adminstration of $10\%,\;20\%$ and $40\%$ BA-noodles compared with control group. These results suggest that administration of BA-noodle can not only effectively inhibit chronic degenerative disease, but also may be intervent the aging process.

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Effect of Brown Algae (Undasia pinnatisda)-Noodle on Inhibitory Action of Obesity in SD Rats (미역 (Undasia pinnatisda) 국수가 SD계 흰쥐의 비만 억제작용에 미치는 영향)

  • CHOI Jin-Ho;KIM Dong-Woo;KIM Jung-Hwa;KIM Dae-Ik;LEE Jong-Soo;BAEK Yeong-Ho
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.32 no.1
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    • pp.46-49
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    • 1999
  • Our previous studies have demonstrated the biological effects of alginic acid as a brown algae (Undasia pinnatisda) component on inhibitory action of obesity using animal model. This study was designed to investigate the effect of brown algae (BA)-noodles ($10\%,\;20\%$ and $40\%$) on inhibitory action of obesity in Sprague-Dawley (SD) male rats for 4 weeks. Body weight gain in SD rats for 4 weeks resulted in a marked inhibition ($7\%,\;17\%\;38\%$, respectively) by administration of $10\%,\;20\%$ and $40\%$ BA-noodles compared with control group. Total energy intake and feed efficiency in SD rats for 4 weeks resulted in a marked inhibition ($6\%,\;16\%\;37\%$ and $10\%\;20\%\;40\%$, respectively) by administration of $10\%\;20\%$ and $40\%$ BA-noodles compared with control group, but gross efficiency was not observed in these BA-noodle groups. Rohrer index and TM index as an obesity markers in SD rats for 4 weeks significantly decreased ($10\%\;15\%$ and $22\%$, respectively) by administration of $10\%,\;20\%$ and $40\%$ BA-noodles compared with control group. These results suggest that inhibitory effect of obesity in SD rats for 4 weeks may be due to decreases of body fat content resulted in a decrease of feed efficiency and energy intake by these BA-noodles.

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Application of the Pulsatile Cardiopulmonary Bypass in Animal Model (이중 박동성 인공심폐기의 동물 실험)

  • Shin, Hwa-Kyun;Won, Yong-Soon;Lee, Jea-Yook;Her, Keun;Yeum, Yook;Kim, Seung-Chul;Min, Byoung-Goo
    • Journal of Chest Surgery
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    • v.37 no.1
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    • pp.1-10
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    • 2004
  • Background: Currently, the cardiopulmonary machine with non-pulsatile pumps, which are low in internal circuit pressure and cause little damage to blood cells, is widely used. However, a great number of experimental studies shows that pulsatile perfusions are more useful than non-pulsatile counterparts in many areas, such as homodynamic, metabolism, organ functions, and micro-circulation. Yet, many concerns relating to pulsatile cardiopulmonary machines, such as high internal circuit pressure and blood cell damage, have long hindered the development of pulsatile cardiopulmonary machines. Against this backdrop, this study focuses on the safety and effectiveness of the pulsatile cardiopulmonary machines developed by a domestic research lab. Material and Method: The dual-pulsatile cardiopulmonary bypass experiment with total extracorporeal circulation was conducted on six calves, Extracorporeal circulation was provided between superior/inferior vena cava and aorta. The membrane oxygenator, which was placed between the left and right pumps, was used for blood oxygenation. Circulation took four hours. Arterial blood gas analysis and blood tests were also conducted. Plasma hemoglobin levels were calculated, while pulse pressure and internal circuit pressure were carefully observed. Measurement was taken five times; once before the operation of the cardiopulmonary bypass, and after its operation it was taken every hour for four hours. Result: Through the arterial blood gas analysis, PCO2 and pH remained within normal levels. PO2 in arterial blood showed enough oxygenation of over 100 mmHg. The level of plasma hemoglobin, which had total cardiopulmonary circulation, steadily increased to 15.87 $\pm$ 5.63 after four hours passed, but remained below 20 mg/㎗. There was no obvious abnormal findings in blood test. Systolic blood pressure which was at 97.5$\pm$5.7 mmHg during the pre-circulation contraction period, was maintained over 100 mmHg as time passed. Moreover, diastolic blood pressure was 72.2 $\pm$ 7.7 mmHg during the expansion period and well kept at the appropriate level with time passing by. Average blood pressure which was 83$\pm$9.2 mmHg before circulation, increased as time passed, while pump flow was maintained over 3.3 L/min. Blood pressure fluctuation during total extracorporeal circulation showed a similar level of arterial blood pressure of pre-circulation heart. Conclusion: In the experiment mentioned above, pulsatile cardiopulmonary machines using the doual-pulsatile structure provided effective pulsatile blood flow with little damage in blood cells, showing excellence in the aspects of hematology and hemodynamic. Therefore, it is expected that the pulsatile cardiopulmonary machine, if it becomes a standard cardiopulmonary machine in all heart operations, will provide stable blood flow to end-organs.