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Protective Effects on A2Kb Transgenic Mice That Were Immunized with Hepatitis B Virus X Antigen Peptides by the Activation of CD8+ T Cells; XEP-3 Specific CTL Responses in the in vitro Culture (B형 간염 바이러스 X 항원을 면역한 A2Kb Transgenic Mice에서 CD8+ T Cell의 활성화에 의한 X 항원 표현 재조합 Vaccinia Virus에 대한 방어 효과; in vitro 배양을 통한 XEP-3 특이적인 CTL의 반응)

  • Hwang, Yu Kyeong;Kim, Hyung-Il;Kim, Nam Kyung;Park, Jung Min;Cheong, Hong Seok
    • IMMUNE NETWORK
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    • v.2 no.1
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    • pp.41-48
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    • 2002
  • Background: Viral antigens presented on the cell surface in association with MHC class I molecules are recognized by CD8+ T cells. MHC restricted peptides are important in eliciting cellular immune responses. As peptide antigens have a weak immunigenicity, pH-sensitive liposomes were used for peptide delivery to induce effective cytotoxic T lymphocyte (CTL) responses. In the previous study, as the HBx peptides could induce specific CTLs in vitro, we tested whether the HLA-A2/$K^b$ transgenic mice that were immunized by HBx-derived peptides could be protected from a viral challenge. Methods: HBx-peptides encapsulated by pH-sensitive liposomes were prepared. $A2K^b$ transgenic mice were immunized i.m. on days one and seven with the indicated concentrations of liposome-encapsulated peptides. Three weeks later, mice were infected with $1{\times}10^7pfu$/head of recombinant vaccinia virus (rVV)-HBx via i.p. administration. The ovaries were extracted from the mice, and the presence of rVV-HBx in the ovaries was analyzed using human TK-143B cells. IFN-${\gamma}$ secretion by these cells was directly assessed using a peptide-pulsed target cell stimulation assay with either peptide-pulsed antigen presenting cells (APCs), concanavalin A ($2{\mu}g/ml$), or a vehicle. To generate peptide-specific CTLs, splenocytes obtained from the immunized mice were stimulated with $20{\mu}g/ml$ of each peptide and restimulated with peptide-pulsed APC four times. The cytotoxic activity of the CTLs was assessed by standard $^{51}Cr$-release assay and intracellular IFN-${\gamma}$ assay. Results: Immunization of these peptides as a mixture in pH-sensitive liposomes to transgenic mice induced a good protective effect from a viral challenge by inducing the peptide-specific CD8+ T cells. Mice immunized with $50{\mu}g/head$ were much better protected against viral challenge compared to those immunized with $5{\mu}g$/head, whereas the mice immunized with empty liposomes were not protected at all. After in vitro CTL culture by peptide stimulation, however, specific cytotoxicity was much higher in the CTLs from mice immunized with $5{\mu}g/head$ than $50{\mu}g/head$ group. Increase of the number of cells that intracellular IFN-${\gamma}$ secreting cell among CD8+ T cells showed similar result. Conclusion: Mice immunized with XEPs within pH-sensitive liposome were protected against viral challenge. The protective effect depended on the amount of antigen used during immunization. XEP-3-specific CTLs could be induced by peptide stimulation in vitro from splenocytes obtained from immunized mice. The cytotoxic effect of CTLs was measured by $^{51}Cr$-release assay and the percentage of accumulated intracellular IFN-${\gamma}$ secreting cells after in vitro restimulation was measured by flow cytometric analysis. The result of $^{51}Cr$-release cytotoxicity test was well correlated with that of the flow cytometric analysis. Viral protection was effective in immunized group of $50{\mu}g/head$, while in the in vitro restimulation, it showed more spectific response in $5{\mu}g$/head group.

The Influence of the Restrictions in Chinese economic growth on Korean commercial environment (중국 경제성장의 제약요인이 한국 통상환경에 미치는 영향)

  • Shong, Il-Ho;Lee, Gye-Young
    • International Commerce and Information Review
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    • v.15 no.4
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    • pp.457-479
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    • 2013
  • Through a Chinese rise, Chinese dream is actualizing as the world's great power. According to outlook of World Bank and IMF, Around 2030 China will be a great power bigger than America's economic power. The rise of China will give a huge impact to the whole world. China expands her influence through a global manufacturing base and a global market. To actualize 'Peaceful Rise' Strategy, China has many constraints. Chinese society is facing many difficult social problem due to side effects of a rapid development. Such as the spread of corruption, the severity of wealth gap, environmental degradation and energy shortage. Internationally there are containment from hegemon so-called 'China threat' dispute, Taiwan issue and territorial disputes. Western countries are hostile to China for two reasons. Based on expectations, one is China's socialist system and the other is the rising China which will compete for supremacy with Europe and America. Recent emergence of Chinese nationalism and the containment of the neighboring countries are also serious limiting factors. Domestically they have the rampant corruption in the bureaucracy, weakened capacity of Communist rule, wealth disparity due to the discriminatory economic development strategy, seriousness of rural problem, social instability, lack of social security systems and the development gap between the eastern coastal areas and western inland areas, ethnic minorities problems, the constraint of sustainable development issues due to lack of resources, environmental pollution and energy constraints. Like the former Soviet Union, China may face a dismantlement. After the rise, China may encounter possibilities of a war between great powers or a collapse of Chinese society caused by deepening internal conflict. Serious economic polarization would make peasants and urban workers, who are social vulnerable people, to turn their back to communist party and threaten the justification and the appropriateness of the ruling communist party. Chinese government will think internal system security threat is more formidable risk factor than a system security threat from the hegemon. The decline of great country comes from internal reasons rather than external reasons. To achieve peaceful rise, unification with Taiwan is an essential prerequisite. Taiwan issues are complex problems which equipped with international and domestic factors. Lack of energy resources, environmental pollution in China will bring economic crisis to Korean enterprises. Important influence to Korean economy will be a changeover of the method in economic development. It will turn the balance of investment and consumption, GDP-centered growth to consumption and environment-centered growth. Services industries including finance, environment, culture, education, health care and social welfare will grow. Change in China's growth model will give a great challenge upon the intermediate goods industry in Korea. Korea should reduce the portion of machinery, automotive, semiconductor, steel and chemical-centered export industry to China, and should increase the proportion of the service industry.

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A Study on Case for Localization of Korean Enterprise in Indonesia (인도네시아 진출 한국기업의 현지화에 관한 사례 연구)

  • Swo, Min-Kyo;Kim, Hee-Jun
    • International Commerce and Information Review
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    • v.15 no.4
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    • pp.481-508
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    • 2013
  • The purpose of this study is to research the specific ways of successful localization by analyzing the success and failures case for localization through the theoretical background and the strategic models of localization. The strategic models of localization are divided by management aspects such as the localization of production and sourcing, the localization of human resources, the localization of marketing, the localization of R&D, harmonious relationship with the local community and authority transfer between headquarters and local subsidiaries. And the specific measures of the successful localization are proposed within the framework of the strategic models by comparing and analyzing the success and failures case for localization of individual companies operating in Indonesia. The results indicate that there are successful companies which develop a suitable products for the local climate and failed automobile company which is weak for assembly of complete vehicle in terms of localization of production and sourcing. In case of localization of human resources, most companies recognize the importance of this part and endeavor to secure superior human resource through a related education. It is found that most of the companies perform R & D in their native country. In part of a harmonious relationship with the local community, Korean companies should contribute to the community and be friendly with local residents and make a good image of the company focusing on the cultural environment. In aspect of authority transfer between headquarters and local subsidiaries, there is a tendency to be determined by the head office rather than the joint participation. In the future, in order for Korean enterprise to be successful one in Indonesia market, a highly interdependent and complex forms between headquarters and local subsidiaries shall be performed and an active exchange of information and the selection of best talent regardless of nationality shall be promoted.

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Experimental Study on the Thermodynamic Characteristics of Commercial Small-size Moxa Combustion (상용 소형 쑥뜸의 열역학적 특성에 대한 실험적 연구)

  • Lee Geon-Mok;Hwang Yoo-Jin;Lee Gun-Hyee
    • Journal of Acupuncture Research
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    • v.18 no.6
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    • pp.171-187
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    • 2001
  • Objective : Moxibustion has been proved efficacious for many diseases, but isn't widespread in the clinics due to a danger of skin burning, the smoke produced while burning a moxa combustion and so on. Therefore, another type of moxa that can be resolved these troubles is required. To improve the effect of moxibustion and develop the new thermal stimulating treatment, the performance of commercial moxibustion widely used are studied systematically and found out quantitatively. Methods : We have selected two types (small-size moxa A(sMA), small-size moxa B (sMB)) among small-size moxaes used widely in the clinic. We examined combustion time, various temperatures, temperature gradient in each period during a combustion of moxa. Results : 1. The combustion time in the preheating period appeared somewhat longer in sMA than in sMB. 2, The combustion time in the heating period appeared longer in sMA by 26% than in sMB. 3. The average temperature in the heating period was $37.6{\sim}37.8^{\circ}C\;in\;sMA\;and\;36.2{\sim}36.8^{\circ}C$ in sMB and the maximum temperature measured at a center of contact surface in sMA was $48.6^{\circ}C$, higher by over $2.8^{\circ}C$ than that of sMB moxibustion. 4. The average ascending temperature gradient in the heating period was $0.08{\sim}0.1^{\circ}C/sec$ in both moxaes, and the average ascending temperature gradient of heating period in sMB appeared larger. The maximum ascending temperature gradient appeared higher in sMB, and the time reaching maximum ascending temperature gradient appeared much earlier in sMA than in sMB. 5. The combustion time in the retaining period was around 100 sec in sMA and around 275 sec in sMB. 6. The average temperature in the retaining period was $42.2{\sim}46.0^{\circ}C\;in\;sMA\;and\;39.3{\sim}41.4^{\circ}C/sec$ in sMB. The minimum temperature in the retaining period was over $38.80^{\circ}C$ in sMA but just $34.7^{\circ}C$ in sMB. 7. The average descending temperature gradient in sMA was $-0.050{\sim}0.067^{\circ}C/sec$ and in sMB was $-0.030{\sim}0.037^{\circ}C/sec$ 8. The combustion time in the cooling period appeared longer over two times in sMA than in sMB, and the time which the cooling period (minimum temperature) finished at appeared later in sMB by 55 sec. 9. We classified the combustion process that the measured temperature rose over body heat($37^{\circ}C$) into the effective combustion period. The effective combustion time was 233.3 sec in sMA and 300.4 sec in sMB respectively, and was longer by about 29% in sMB. The average temperature and maximum temperature in the effective combustion time appeared higher in sMA. The time taken until the maximum temperature was reached was 225.1 sec in sMA and 244.5 sec in sMB, faster by about 20 sec in sMA. The maximum ascending temperature gradient during the effective combustion period appeared larger about 1.4 times in sMB, but the time when the maximum ascending temperature gradient happened was faster in sMA. Conclusion : It appears that sMB, compared with sMA, is proper if necessary to apply the long time and weak stimulus, because of the gentle stimulus during the relatively longer time. In contrast, sMA that the symmetrical combustion happened is proper if necessary to apply the short time and strong stimulus.

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Analysis for the Idiopathic Facial Palsy Inpatients According to Distribution of Sasang Constitution, Hyungsang Classification and Assessment Tools (특발성 안면신경마비 환자에 대한 사상체질.형상별 분포 및 평가도구에 따른 분석)

  • Lee, Seung Hwon;Lee, Eun Sol;Seo, Dong Kyun;Lee, Kyeong A;Kim, Jung Hee;Hong, Chang Ho;Jang, Sun Hee;Youn, Hyoun Min;Jang, Kyung Jeon;Song, Choon Ho;Kim, Cheol Hong
    • Journal of Acupuncture Research
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    • v.30 no.4
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    • pp.55-68
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    • 2013
  • Objectives : The purpose of this study is to analyze the distribution, relationship, prognosis and improvement score of idiopathic facial palsy inpatients according to constitutional differentiation ; Sasang constition, Hyungsang classification, Sasang constition combined with Hyungsang classification. Methods : A study was done on 102 patients who were diagnosed and treated as idiopathic facial palsy from April 2012 to Nomember 2012 at the Department of Acupuncture and Moxibution Medicine, Dong-eui Oriental University Hospital. Medical records of inpatinets who underwent facial ENoG, NET test were analyzed. Changes of HBGS grade and Yanagihara's score were also evaluated. We classified inpaients acording to Sasang constitution by consulting to Department of Sasang constitutinal medicine, and to Hyungsang classification(Dam-body, Bangkwang-body) by measuring under medical system of 3D facial shapes(RS-400FL). The constitutional differentiation and general characteristics were initially analyzed, and valuse on ENoG, NET were correlated with constitutinal differentiation. Results of HBGS and Yanagihara's score were also correlated with constitutinal differentiation. Results : 1. Each 39.2 percent of idiopathic facial palsy inpatients were the Taeeumin and Soyangin, 21.6 percent were the Soeumin. 2. 75.5 percent of idiopathic facial palsy inpatients were the Bangkwang-body, 24.5 percent were the Dam-body. 3. 34.3 percent of idiopathic facial palsy inpatinets were the Taeeum-Bangkwang, 21.6 percent were the Soyang-Bangkwang, 19.6 percent were the Soeum-Bangkwang, 17.6 percent were the Soyang-Dam, 4.9 percent were the Taeem-Dam, 2 percent were the Soeum-Dam. 4. By sex, the each ratio of Taeeumin, Soeumin, Bangkwang-body, Taeeum-Bangkwang, Soeum-Bangkwang, Soyang-Bangkwang was higher in female, that of Dam-body, Taeeum-Dam, Soyang-Dam was higher in male. 5. By relations between Sasang constitution and Hyungsang classification, the each ratio of Taeeumin and Soeumin was higher in Bangkwang-body. 6. By values on ENoG and NET, evaluations of HBGS's grade and Yanagihara's score, there were no significant difference. Conclusions : In idiopathic facial palsy inpatients, the proportion of Taeeum and Soyang among the Sasang constitution was higher, that of Dam among the Hyungsang classification was higher, that of Taeeum-Bangkwang among the Sasang combined with Hyungsang was the highest. It would seem that Bangkwang-body, female were closely related to Taeeumin, Soeumin. Also, Dam-body were closely related to male. But when comparing groups, there was no statistically significant difference in prognosis and improvement.

A new diagnostic method for diabetic neuropathy using $TcpO_2$ ($TcpO_2$ 이용한 당뇨병성 신경병증 환자의 조기진단)

  • Hong, Hyun-Ki;Kim, Sung-Woo;Nam, Ki-Chang;Cha, Eun-Jong;Kim, Deok-Won
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.44 no.3
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    • pp.1-8
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    • 2007
  • PDiabetic neuropathy is one of the most common diabetes related complications including diabetic nephropathy and retinopahty. In clinical practices, nerve conduction velocity (NCV) has been used as a standard method for diagnosing diabetic neuropathy. However, it applies maximum current of 100mA to nerves causing stress and pain to patients. In this study, as a non-invasive method, $TcpO_2$ was utilized to investigate the difference and relationship between $TcpO_2$ and $SpO_2$ of normal and diabetic neuropathy subjects. In addition, a new method of diagnosing diabetic neuropathy using $TcpO_2$ is suggested. 50 normal subjects and 50 diabetic patients with neuropathy diagnosed by NCV participated in this study. Parameters used in this study were $TcpO_2$, $TcpCO_2$, and $SpO_2$. As a result of the $TcpO_2$ measurements, statistical significances were found from $TcpO_2$ of hands and feet from normal and patients group(p<0.01). $SpO_2$ measured from index finger of normal and patient groups showed no statistical significance(p>0.05). On the other hand, $SpO_2$ measured from great toes of normal and patient groups showed statistical significance(p<0.01). Correlation coefficient between $SpO_2$ of finger and $TcpO_2$ of hand was 0.400 (p<0.01) and $SpO_2$ of toe and $TcpO_2$ of foot was 0.471(p<0.01). Both correlation values were statistically significant. Sensitivities and specificities of the $TcpO_2$ method were found to be 66 % and 92 %, respectively. If the suggested $TcpO_2$ method is used periodically, prevention and early diagnosis of diabetic neuropathy would be possible.

Effects of antler, red ginseng, safflower, ipriflavone and estrogen on body weight, bone mineral density, organ weight and histological change in ovariectomized rats (녹용, 홍삼, 홍화, ipriflavone 및 estrogen이 난소적출 rat의 체중, 골밀도, 장기중량 및 조직학적 변화에 관한 연구)

  • 유상식;김상근;이명헌;송운재;문병천;김무강
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.235-245
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    • 2000
  • This study was peformed to elucidate the effects of antler, red ginseng, safflower seed, ipriflavone and estrogen on ovariectomized rats. The rats were fed with Ca and P deficient diet for five weeks to induce osteoporosis. After this period, these animals were fed with normal for and treated every other day with antler(600mg/kg, PO), red ginseng(200mg/kg, PO), safflower(200mg/kg, PO), ipriflavone(80mg/kg, PO) and estrogene(400$\mu\textrm{g}$/kg, IM) for S weeks. During the treatment, the rats were examined for body, organ, femur and femur ash weights. And electron-microscopical examinations were also performed on femora. The results are summarized as follows : 1. The body weights of the antler, red ginseng, safflower, ipriflavone and estrogen-treated groups after five weeks of treatment, were 303.0$\pm$3.0g, 273.3$\pm$4.4g, 288.3$\pm$5.1g, 300.5$\pm$5.2g, 320.5$\pm$4.2g, respectively. They, except the red ginseng-treated group, were significantly higher than 278.4$\pm$2.7g of normal control group(P<0.05). And the body weight of non-treatment group was 29it.2$\pm$5.5g which was higher than that of control group, too. 2. The weights of left femur after five weeks of treatment were showed 0.83$\pm$0.2g by antler, 0.89$\pm$0.2g by red ginseng, 0.87$\pm$0.2g by safflower 0.83$\pm$0.1g by ipriflavone and 0.78$\pm$0.1g by estrogen-treated group. They were a little lower than 0.97$\pm$0.1g of normal control group. Non-treatment group showed 0.83$\pm$0.1g which was lower than that of control group but not significant. 3. The weights of ashed left femora after five weeks of treatment were showed 0.36$\pm$0.01g by antler, 0.40$\pm$0.01g by red ginseng, 0.41$\pm$0.01g by safflower, 0.36$\pm$0.01g by ipriflavone and 0.35$\pm$0.01g by estrogen-treated group. Those results were significantly lower than 0.43$\pm$0.01g of normal control group. And non-treatment group showed 0.29$\pm$0.01g of femoral ash which was significantly lower than that showed by control group(p<0.05). 4. The femoral Ca contents of the antler, red ginseng, safflower, ipriflavone and estrogen-treated rats were 19.6$\pm$0.7%, 17.8$\pm$1.1%, 18.8$\pm$0.9%, 18.0$\pm$1.1%, 15.3$\pm$1.1%, respectively, after five weeks of treatment. The femoral Ca contents of antler and safflower-treated groups were higher than that of control group which showed 18.2$\pm$0.9% of Ca The non-treatment group, showing 15.3$\pm$0.7%, had significantly lower content of Ca than normal group(p<0.05). 5. The weights of liver, spleen, and kidney in the osteoporosis-induced rats during the treatment with antler, red ginseng, safflower, jpriflavone and estrogen showed not significant changes. 6. The connections among the trabeculae of control group were maintained well, showing no bone loss. However, the connections among the trabeculae of non-treatment group were thinner than those of control group and were almost disconnected and the lacunae were found to be broadened. Antler and safflower-treated group showed only slight changes. The recovery rates were prominent in antler, safflower, estrogen-treated groups but they were insignificant in red ginseng and ipriflavone-treated groups.

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An Experimental Study on Flow Characteristics for Optimal Spacing Suggestion of 45° Upward Groynes (45° 상향수제의 적정 간격 제시를 위한 흐름특성 실험 연구)

  • Kim, Sung Joong;Kang, Joon Gu;Yeo, Hong Koo
    • Journal of Korea Water Resources Association
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    • v.47 no.5
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    • pp.459-468
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    • 2014
  • Groyne to control the direction and velocity of flow in rivers is generally installed for the purpose of protecting riverbanks or embankments from erosion caused by running water. In particular, as interest in river restoration and natural river improvement increases, groynes are proposed as a key hydraulic structure for local flow control and riparian habitat establishment. Groynes are installed mainly in groups rather than as individual structures. In case of groynes installed as a group, flow around the groynes change according to spacing in between the groynes. Therefore, groyne spacing is regarded as the most important factor in groyne design. This study aimed at examining changes of flows around and within the area of groynes that take place according to the spacing of groynes installed in order to propose the optimal spacing for upward groynes. To examine flow characteristics around groynes, this study looked at flows in main flow area and recirculation flow area separately. In main flow area, it examined the impact of flow velocity increasing as a result of conveyance reduction that is exerted on river bed stability in relation to changes in the maximum flow velocity according to installation spacing. As a factor causing impacts on scouring and sedimentation within the area of groynes, recirculation flow in the groyne area can lead problems concerning flow within the area and stability of embankment. As for recirculation area, an analysis was conducted on the scale of rotational flow and the flow around embankment that exerts impacts on stability of the embankment. In addition, a comparative analysis was carried with reference to changes of the central point of rotational flow that occur within the area of groynes. As a result of compositely examining the results, the appropriate installation spacing is proposed as min. four times-max. six times considering a decrease in flow velocity according to the installation of upward groynes, river bed stability and stability of embankments against counterflow within the area of groynes.

Developmental competence and Effects of Coculture after Crypreservation of Blastomere-Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis (착상 전 유전진단 기술 개발의 동물실험 모델로서 할구 생검된 생쥐 배아에서 동결보존 융해 후 배아 발생 양상과 공배양 효과에 관한 연구)

  • Kim, Seok-Hyun;Kim, Hee-Sun;Ryu, Buom-Yong;Choi, Sung-Mi;Pang, Myung-Geol;Oh, Sun-Kyung;Jee, Byung-Chul;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Moon, Shin-Yong;Lee, Jin-Yong;Chae, Hee-Dong;Kim, Chung-Hoon
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.47-57
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    • 2000
  • Objective: The effects of cryopreservation with or without coculture on the in vitro development of blastomere-biopsied 8-cell mouse embryos were investigated. This experimental study was originally designed for the setup of a preclinical mouse model for the preimplantation genetic diagnosis (PGD) in human. Methods: Eight-cell embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BL(표현불가)/CBA(표현불가)). Using micromanipulation, one to four blastomeres were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acid Tyrode's solution (ATS). A slow-freezing and rapid-thawing protocol with 1.5M dimethyl sulfoxide (DMSO) and 0.1M sucrose as cryoprotectant was used for the cryopreservation of blastomere- biopsied 8-cell mouse embryos. After thawing, embryos were cultured for 110 hours in Ham's F-10 supplemented with 0.4% bovine serum albumin (BSA). In the coculture group, embryos were cultured for 110 hours on the monolayer of Vero cells in the same medium. The blastocyst formation was recorded, and the embryos developed beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. Results: The survival rate of embryos after cryopreservation was significantly lower in the blastomere-biopsied (7/8, 6/8, 5/8, and 4/8 embryos) groups than in the non-biopsied, zona intact (ZI) group. Without the coculture, the blastocyst formation rate of embryos after cryopreservation was not significantly different among ZI, the zona drilling only (ZD), and the balstomere-biopsied groups, but it was significantly lower than in the non-cryopreserved control group. The mean number of cells in embryos beyond blastocyst stage was significantly higher in the control group ($50.2{\pm}14.0$) than in 6/8 ($26.5{\pm}6.2$), 5/8 ($25.0{\pm}5.5$), and 4/8 ($17.8{\pm}7.8$) groups. With the coculture using Vero cells, the blastocyst formation rate of embryos after cryopreservation was significantly lower in 5/8 and 4/8 groups, compared with the control, 7/8, and 6/8 groups. The mean number of cells in embryos beyond blastocyst stage was also significantly lower in 4/8 group ($25.9{\pm}10.2$), compared with the control ($50.2{\pm}14.0$), 7/8 ($56.0{\pm}22.2$), and 6/8 ($55.3{\pm}25.5$) groups. Conclusion: After cryopreservation, blastomere-biopsied mouse embryos have a significantly impaired developmental competence in vitro, but this detrimental effect might be prevented by the coculture with Vero cells in 8-cell mouse embryos biopsied one or two blastomeres. Biopsy of mouse embryos after ZD with ATS is a safe and highly efficient preclinical model for PGD of human embryos.

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Study on the Vitrification of Human Blastocysts: II. Effect of Vitrification on the Implantation and the Pregnancy of Human Blastocysts (인간 포배기 배아의 초자화 동결에 관한 연구: II. 초자화 동결이 포배기 배아의 착상 및 임신에 미치는 영향)

  • Kim, Su-Hee;Lee, Sang-Won;Lee, Ju-Hee;Kang, Sang-Min;Oh, Hee-Jeong;Lee, Seoung-Min;Lee, Seong-Goo;Yoon, Hye-Gyun;Yoon, San-Hyun;Park, Se-Pill;Song, Hai-Bum;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.1
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    • pp.67-74
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    • 2000
  • Objective: This study was conducted to investigate the effect of vitrification on the implantation and the pregnancy of human blastocysts. Method: The transfer of the frozen-thawed blastocysts by the slow freezing or vitrification was performed between January 1998 and July 1999. The zygotes derives from IVF were cocultured with cumulus cells in YS medium containing 20% hFF for 5days. Two or three of the best balstocysts produced on day 5 were transferred into the uterus, and then supernumerary blastocysts were randomly divided into two groups. One was frozen by slow freezing and the other was frozen by vitrification method. The slow freezing procedure was performed in two steps (5% glycerol and 9% glycerol + 0.2 M sucrose for 10 min, respectively) using programmed freezer ($-2^{\circ}C$/min to $-7^{\circ}C$, manual seeding at $-7^{\circ}C$, $-0.3^{\circ}C$/min to $-38^{\circ}C$ and plunged into $LN_{2}$). The blastocysts frozen by slow freezing were thawed at $36^{\circ}C$ then removed glycerol in 7 steps. The vitrification procedure was performed in three steps (10% glycerol for 5 min, 10% glycerol + 20% ethylene glycol for 5 min, 25% glycerol + 25% ethylene glycol and directly $LN_{2}$ within 1 min). The blastocysts frozen by vitrification were thawed at $20^{\circ}C$ water then removed cryoprotectant in 3 steps. In each group, thawed blastocysts were cocultured with cumulus cells in YS medium containing 20% hFF for 18h and transferred into the uterus. The implantation rate was evaluated per transferred blastocysts and the pregnancy rate was evaluated per transfers. Results: The survival rate of vitrified group (74.5%) was higher than slow freezing group (68.0%), but not significant. When 98 thawed blastocysts of vitrification were transferred in 40 cycles, 19 pregnancies (clinical pregnancy rate; 47.5%) were established. One miscarriage occurred in the eighth week of pregnancy (ongoing pregnancy rate; 45.0%). 7 pregnancies were ongoing, 11 pregnancies went to term, and 16 healthy infants were born. The Implantation rate was 31.6%. These results were higher than those obtained by the slow freezing (clinical pregnancy rate; 40.3%, ongoing pregnancy rate; 32.5% and implantation rate; 25.3%), but not significant. Conclusion: Vitrification is a simple, quick and economical method when compared to slow freezing. It will be chosen as a good method of human embryo freezing in IVF-ET programs.

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