• Analytical Science and Technology
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• v.21 no.2
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• pp.93-101
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• 2008

The application of solid phase microextraction (SPME) is generally conducted by directly immersing the fiber into the liquid sample or by exposing the fiber in the head space (HS). The extraction temperature, the time of incubation, and application of stirring are often designated to be the most important parameters for achieving the best extraction efficiencies of HS-SPME analysis. In this study, relative importance of these three analytical parameters involved in the HS-SPME method is evaluated using a polydimethylsiloxane/carboxen (PDMS/CAR) fiber. To optimize its operation conditions the competing relationships between different parameters were investigated by comparing the extraction efficiency based on the combination of three parameters and two contracting conditions: (1) heating the sample at 30 vs. 50 C, (2) exposing samples at two durations of 10 vs. 30 min, and (3) application of stirring vs. no stirring. According to our analysis among 8 combination types of HS-SPME method, an extraction condition termed as S50-30 condition ((1) 1200 rpm stirring, (2) $50^{\circ}C$ exposure temp, and (3) 30 min exposure duration) showed maximum recovery rate of 45.5~68.5% relative to an arbitrary reference of direct GC injection. According to this study, the employment of stirring is the most crucial factor to improve extraction efficiency in the application of HS-SPME.

• Development and Reproduction
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• v.15 no.2
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• pp.179-185
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• 2011

Some phytoestrogens in soy and red wine, for example, might have beneficiary rather than adverse effects. In particular, dietary soy intake seems to be highly correlated with protection of breast cancer, osteoporosis and cardiovascular disorders. However, questions persist on the potential adverse effects of the main soy constituent genistein (GS) on female reproductive physiology. Previously we found that prepubertal exposure to GS could activate the reproductive system of immature female rats leading to precocious puberty onset, and intracerebroventricularly (ICV) injected GS could directly activate hypothalamic kisspeptin-GnRH neuronal circuits in adult female rats. The present study was performed to examine the hypothalamus-specific GS effects in prepubertal female rats and which subtype of estrogen receptor is mediated in this GS effect. Prepubertal female rats (PND 30) were anaesthetized, treated with single dose of GS (3.4 ${\mu}g$/animal), and sacrificed at 2 hrs post-injection. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus, total RNAs were extracted and applied to the semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). ICV infusion of GS significantly lowered the transcriptional activities of mTOR (1:$0.361{\pm}0.058$ AU, p<0.001) but increased that of GAD67 (1:$1.285{\pm}0.099$ AU, p<0.05), which are known to act as an upstream modulator of kisspeptin and GnRH neuronal activities in the hypothalamus, respectively. GS administration enhanced significantly the mRNA levels of KiSS-1(1:$1.458{\pm}0.078$ AU, p<0.001), and exerted no effect on the mRNA level of kisspeptin receptor GPR-54 (1:$1.29{\pm}0.08$ AU). GnRH gene expression was significantly decreased in GS-treated group compared to control group (1:$0.379{\pm}0.196$ AU, p<0.05). There was no difference in the mRNA level of $ER{\alpha}$ in the GS-treated group compare to control group (1:$1.180{\pm}0.390$ AU, Fig. 3A). However, icv infusion of GS significantly increased the transcriptional activities of $ER{\beta}$ (1:$4.209{\pm}0.796$ AU, p<0.01, Fig. 3B). Taken together, the present study indicated that the acute exposure to GS could directly alter the hypothalamic GnRH modulating system in prepubertal female rats. Our study strongly suggested the involvement of $ER{\beta}$ pathway in GS's hypothalamus-specific action, and this idea is consistent with the GS's well-known $ER{\beta}$-mediated protective action in breast cancer.

• The Journal of Korean Medicine
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• v.18 no.1
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• pp.480-498
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• 1997

This study was to investigate the protective and anticirrhotic effects of Mockhyangjokisan and Haewooljoweetang on the liver cirrhosis or fibrosis induced by prolonged bile duct ligation; a new experimental model for cirrhosis and the intraperitoneal injection of dimethylnitrosamine in the rat. The development of fibrosis or cirrhosis and its inhibition by the two prescriptions were examined by the chemical analysis of AST, ALT, and hydroxyproline. The results obtained were as follows. 1. The increase of serum asparate aminotransferase induced by bile duct ligation was inhibited by the administration of Mockhyangjokisan and Haewooljoweetang extract. 2. The increase of serum alanine aminotransferase induced by bile duct ligation was inhibited by the administration of Mockhyangjokisan and Haewooljoweetang extract. 3. The increased level of serum AST and AL T induced by the intraperitoneal injection of dimethylnitrosamine was inhibited by the administration of Mockhyangjokisan and Haewooljoweetang extract. 4. The increasing level of hydroxyproline volume in the damaged liver tissues in the rat was decreased by the oral administration of Mockhyangjokisan and Haewooljoweetang extract. But there were no significant differences in the inhibition rate between the two experimental groups.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.5
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• pp.375-395
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• 2006

Purpose: Lingual nerve (LN) damage may be caused by either tumor resection or injury such as wisdom tooth extraction, Although autologous nerve graft is sometimes used to repair the damaged nerve, it has the disadvantage of necessity of another operation for nerve harvesting. Moreover, the results of nerve grafting is not satisfactory. The nerve growth factor (NGF) is well-known to play a critical role in peripheral nerve regeneration and its local delivery to the injured nerve has been continuously tried to enhance nerve regeneration. However, its application has limitations like repeated administration due to short half life of 30 minutes and an in vivo delivery model must allow for direct and local delivery. The aim of this study was to construct a well-functioning $rhNGF-{\beta}$ adenovirus for the ultimate development of improved method to promote peripheral nerve regeneration with enhanced and extended secretion of hNGF from the injured nerve by injecting $rhNGF-{\beta}$ gene directly into crush-injured LN in rat model. Materials and Methods: $hNGF-{\beta}$ gene was prepared from fetal brain cDNA library and cloned into E1/E3 deleted adenoviral vector which contains green fluorescence protein (GFP) gene as a reporter. After large scale production and purification of $rhNGF-{\beta}$ adenovirus, transfection efficiency and its expression at various cells (primary cultured Schwann cells, HEK293 cells, Schwann cell lines, NIH3T3 and CRH cells) were evaluated by fluorescent microscopy, RT-PCR, ELISA, immunocytochemistry. Furthermore, the function of rhNGF-beta, which was secreted from various cells infected with $rhNGF-{\beta}$ adenovirus, was evaluated using neuritogenesis of PC-12 cells. For in vivo evaluation of efficacy of $rhNGF-{\beta}$ adenovirus, the LNs of 8-week old rats were exposed and crush-injured with a small hemostat for 10 seconds. After the injury, $rhNGF-{\beta}$ adenovirus($2{\mu}l,\;1.5{\times}10^{11}pfu$) or saline was administered into the crushed site in the experimental (n=24) and the control group (n=24), respectively. Sham operation of another group of rats (n=9) was performed without administration of either saline or adenovirus. The taste recovery and the change of fungiform papilla were studied at 1, 2, 3 and 4 weeks. Each of the 6 animals was tested with different solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) by two-bottle test paradigm and the number of papilla was counted using SEM picture of tongue dorsum. LN was explored at the same interval as taste study and evaluated electro-physiologically (peak voltage and nerve conduction velocity) and histomorphometrically (axon count, myelin thickness). Results: The recombinant adenovirus vector carrying $rhNGF-{\beta}$ was constructed and confirmed by restriction endonuclease analysis and DNA sequence analysis. GFP expression was observed in 90% of $rhNGF-{\beta}$ adenovirus infected cells compared with uninfected cells. Total mRNA isolated from $rhNGF-{\beta}$ adenovirus infected cells showed strong RT-PCR band, however uninfected or LacZ recombinant adenovirus infected cells did not. NGF quantification by ELISA showed a maximal release of $18865.4{\pm}310.9pg/ml$ NGF at the 4th day and stably continued till 14 days by $rhNGF-{\beta}$ adenovirus infected Schwann cells. PC-12 cells exposed to media with $rhNGF-{\beta}$ adenovirus infected Schwann cell revealed at the same level of neurite-extension as the commercial NGF did. $rhNGF-{\beta}$ adenovirus injected experimental groups in comparison to the control group exhibited different taste preference ratio. Salty, sweet and sour taste preference ratio were significantly different after 2 weeks from the beginning of the experiment, which were similar to the sham group, but not to the control group.

• The Korean Journal of Emergency Medical Services
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• v.15 no.3
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• pp.31-44
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• 2011

Purpose : The purpose of emergency medical services(EMS) is to save human lives and assure the completeness of the body in emergency situations. Those who have been qualified on medical practice to perform such treatment as there is the risk of human life and possibility of major physical and mental injuries that could result from the urgency of time and invasiveness inflicted upon the body. In the emergency medical activities, 119 emergency medical technicians mainly perform the task but they are not able to perform such task independently and they are mandatory to receive medical direction. The purpose of this study is to examine the recognition and request for medical direction by 119 emergency medical technicians in order to provide basic information on the development of medical direction program suitable to the characteristics of EMS as well as for the studies on EMS for the sake of efficient operation of pre-hospital EMS. Method : Questionnaire via e-mail was conducted during July 1-31, 2010 for 675 participants who are emergency medical technicians, nurses and other emergency crews in Gyeongbuk. The effective 171 responses were used for the final analysis. In regards to the emergency medical technicians' scope of responsibilities defined in Attached Form 14, Enforcement regulations of EMS, t-test analysis was conducted by using the means and standard deviation of the level of request for medical direction on the scope of responsibilities of Level 1 & Level 2 emergency medical technicians as the scale of medical direction request. The general characteristics, experience result, the reason for necessity, emergency medical technicians & medical director request level, medical direction method, the place of work of the medical director, feedback content and improvement plan request level were analyzed through frequency and percentage. The level of experience in medical direction and necessity were analyzed through ${\chi}^2$ test. Results : In regards to the medical direction experience per qualification, the experience was the highest with 53.3% for Level 1 emergency medical technicians and 80.3% responded that experience was helpful. As for the recognition on the necessity of medical direction, 71.3% responded as "necessary" and it turned out to be the highest of 76.9% in nurses. As for the reason for responding "necessary", the reason for reducing the risk and side-effects from EMS for patients was the largest(75.4%), and the reason of EMS delay due to the request of medical direction was the highest(71.4%) for the reason for responding "not necessary". In regards to the request level of the task scope of emergency medical technicians, injection of certain amount of solution during a state of shock was the highest($3.10{\pm}.96$) for Level 1 emergency rescuers, and the endotracheal intubation was the highest($3.12{\pm}1.03$) for nurses, and the sublingual administration of nitroglycerine(NTG) during chest pain was the highest($2.62{\pm}1.02$) for Level 2 emergency medical technicians, and regulation of heartbeat using AED was the highest($2.76{\pm}.99$) for other emergency crews. For the revitalization of medical direction, the improvement in the capability of EMS(78.9%) was requested from emergency crew, and the ability to evaluate the medical state of patient was the highest(80.1%) in the level of request for medical director. The prehospital and direct medical direction was the highest(60.8%) for medical direction method, and the emergency medical facility was the highest(52.0%) for the placement of medical director, and the evaluation of appropriateness of EMS was the highest(66.1%) for the feedback content, and the reinforcement of emergency crew(emergency medical technicians) personnel was the highest(69.0%) for the improvement plan. Conclusion : The medical direction is an important policy in the prehospital EMS activity because 119 emergency medical technicians agreed the necessity of medical direction and over 80% of those who experienced medical direction said it was helpful. In addition, the simulation training program using algorithm and case study through feedback are necessary in order to enhance the technical capability of ambulance teams on the item of professional EMS with high level of request in the task scope of emergency medical technicians, and recognition of medical direction is the essence of the EMS field. In regards to revitalizing medical direction, the improvement of the task performance capability of 119 emergency medical technicians and medical directors, reinforcement of emergency medical activity personnel, assurance of trust between emergency medical technicians and the emergency physician, and search for professional operation plan of medical direction center are needed to expand the direct medical direction method for possible treatment beforehand through the participation by medical director even at the step in which emergency situation report is received.

• Journal of Korean Academy of Nursing
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• v.3 no.3
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• pp.97-110
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• 1973

The goal of modern nursing is to provide comprehensive nursing care to patients. If comprehensive nursing care to children (within the hospital setting) is to be provided, consideration of the stage of growth and development of the child is especially important. From clinical observation, it appeared that nurses often disregarded individual requirements of children in giving nursing care. Therefore, the purpose of this study is to show that comprehensive nursing care which is based on an understanding of the growth and development of the child contributes to both the child and the mother's adaptability to the child's hospitalization. Method: Sixty children, three to three year of age, hospitalized at the Yonsei University Pediatric Ward ware studied. From April 1, 1973 to May 5, 1973, children admitted to the hospital were assigned to either an experimental or a compare groups. There were 30 children in each group. The sex and age of the children in each group was similar. In both groups were more male than female children. In the experimental group, each mother stayed with hot child continuously during his hospitalization. In the compare groups, the mother or some other member of the family stayed with the child. Each day on the child's admission the investigator visited the ward from 1-2 P.M. to 9-10 P.M., in order to provide comprehensive care for the experimental -group. The assistance given the nurses by the investigator was in the form of conferences regarding care and in giving direct care to the child and his mother. The compare group of children received nursing care as usually provided by the hospital. The instruments used to obtain the data for analysis were as follows: 1. The fear and anxiety reaction of the child was recorded by observation of the investigator for four areas: 1) separation from parent and relatives 2) reaction to Doctor and Nurse with white gowns 3) reaction to nursing care 4) reaction to injection and tests, etc. 2. Regression in area of eating, sleeping, and elimination were recorded by the investigator by questioning the mother and by observation. 3. Adaptability to the hospitalization was recorded by direct questioning of the children for areas of emotional and social adjustment. For children older than 3 years of age or children not seriously ill, using the simple I. Q. test this was possible for only 35 of the total 60 children. Result: 1. 55 percents of the total 60 children had been prepared by their parents for hospitalization. The children who had received prior preparation accepted hospitalization more readily than those who had received no preparation. (χ²=4.6 Ρ<0.05) 2. On admission 31.7 percent of the children expressed verbal fear of their discase or treatment. 25 percent felt that the disease was due to their mistake. 3. There was a significant difference in the reaction of the child to separation from the parent or relatives between the two groups. The experimental groups showed less anxiety due to separation than the compare group. (χ²=4.34 Ρ<0.05) In both groups there was less anxiety due to separation among school age (6-12 years) children than among preschool age (3-5 years) children. (χ²=9.22 Ρ<0.05) 4. More than half of the children in both groups reacted with fear and avoidance to doctor and/or nurses wearing white gowns. (χ²=0.06 Ρ<0.05) 5. The experimental group reacted more favorably to nursing in general than the compare group. (χ²=4.8 Ρ<0.05) 6. There was no difference in the fear and refused reaction to special tests and/or such as X-rays and injections, etc. between the groups. (χ²=3.77 Ρ<0.05) 7. More children in the compare group showed regressive tendencies in eating, sleeping, and elimination habits than in experimental groups. (χ²=2.3 Ρ<0.05 χ²=3.88 Ρ<0.05 χ²=4.9 Ρ<0.05) 8. There was a significant difference in the adaptability to hospitalization between the two groups. The experimental groups adapted more readily. (χ²=2.02 Ρ<0.05) 9. For children who had higher I.Q. s the adaptability to hospitalization was better regardless of the group. (χ²=5.03 Ρ<0.05) However, because of the small number of cases (60), this finding cannot be extrapolated without further verification. The date demonstrates that there was a greater adaptability to hospitalization by the child when comprehensive nursing care was given. By planning care and applying knowledge of growth and development to meet, nurses are in a position to prevent some of the psychological trauma associated with hospitalization.

• The Journal of the Korean bone and joint tumor society
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• v.11 no.1
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• pp.46-53
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• 2005

Purpose: Bisphosphonates (BPs) are the analogues of endogenous pyrophosphates: they have been used in the treatment of skeletal diseases such as Paget's disease, osteoporosis, and tumorinducing ostelysis, and are used in treatment of osteolytic metastasis of breast cancer recently. They are also used as one of the therapeutic agents for metastasis of prostatic cancer of which metastasis makes the mixed nature of osteolysis and ostegenesis. Although the action mechanism of BPs are well known for diseases with excessive osteoclastic bone resorption, the direct effect of BPs has not been known yet. This study was intended to see the tumor suppression capability of Zoledronic acid(ZOL) using nude mouse with osteosarcoma. Materials and Methods: MG-63 and HOS osteosarcoma cell lines were used and the transforemed MG-63-GFP and HOS-GFP cells, which were made for detection under fluorescent light, were subcutaneously injected to make osteosarcoma. The five 6-week male mice were used for the experiment at each group. After the injection, mice were cultivated until tumor pieces grow up to $3{\times}3{\times}3$ $mm^3$ and ZOL of 120 ug/kg was subcutaneously injected twice a week. Sizes of tumor were measured twice a week and photographed under fluorescent light. Results: In in vivo test with HOS osteosarcoma cell lines, mean size of tumors was 2,520 $mm^3$ in control group and was 131 $mm^3$ in ZOL group, which showed 94% of reduction comparing with the control ; with MG-63 osteosarcoma cell lines, mean size of tumors was 2,866 $mm^3$ in control group and was 209 $mm^3$ in test group with 72% of reduction (p<0.05). Conclusion: In in vivo tests with nude mice, we suggest that ZOL has direct effect on osteosarcoma cells and it would be used as one of the therapeutic agents for osteosarcoma, especially to ZOL-sensitive osteosarcoma cells.

• Applied Microscopy
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• v.36 no.3
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• pp.173-182
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• 2006

Recently, the pomegranate seed oil (PSO) has been reported to have various efforts including anti-cancer effect. In this study, we examined the liver-protecting effect of the PSO on the hepatotoxicity induced by $CCl_4$ using the BALB/c mice. The PSO was made from dried seeds of black pomegranate (Punica grantum) by heating and squeezing. The expreimental animals were divided into 3 groups; control group treated with olive oil only, experimental group 1 treated with $CCl_4$ only, and experimental group 2 treated with PSO and $CCl_4$. 24 hours after injection of $CCl_4$ into the peritoneal cavity, we collected the blood samples to measure the level of serological factors; aspartate aminotransferase(AST), alanine aminotransferase (ALT), total protein, albumin, total bilirubin, direct bilirubin and alkaline phosphatase. Simultaneously we observed the histological change of liver under the light and electron microscope. As the result, AST and ALT showed $88.7{\pm}14.9IU/L\;and\;22.0{\pm}3.12IU/L$ in the control group, $1963.7{\pm}1212.9IU/L\;and\;4495.4{\pm}2803.6IU/L$ in the experimental group 1, and $432.2{\pm}260.1IU/L\;and\;692.3{\pm}433.1IU/L$ in the experimental group 2. The experimental group 2 showed significant difference as compared with experimental group 1 (P<0.005). In histological study, the experimental group 2 was recovered than experimental group 1 which had abnormal mitochondria, increase of lysosomes, and severe necrosis at the central vein zones. These results indicated that the PSO had the liver protecting effect. However, The further study on the relationship between ingredients of pomegranate seed and liver protecting effect is in need.

• Journal of Food Hygiene and Safety
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• v.38 no.5
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• pp.319-331
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• 2023

Aminoglycoside antibiotics, also known as aminoglycosides (AGs), are veterinary drugs effective against a wide range of gram-negative and gram-positive bacteria. Owing to their recent use in cultured meats, it has become essential to establish an analytical method for safety management. AGs are highly polar compounds, and ion-pair reagents (IPRs) are used to ensure component separation. Owing to the high possibility of potential mechanical problems resulting from IPR addition to the mobile phase, an analytical method in which IPRs are added directly to the vial was explored. In this study, methods for analyzing 10 AGs via liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the addition of two IPRs were validated for selectivity, detection limit, quantitation limit, recovery, and precision. The detection limit was 0.0001-0.0038 mg/kg, the quantification limit was 0.004-0.011 mg/kg, and the linearity (R²) within the concentration range of 0.01-0.5 mg/kg was over 0.99. Recovery and precision (expressed as relative standard deviation) evaluated in the two matrices (beef and cell culture media) ranged from 70.7% to 120.6% and 0.2% to 24.7%, respectively. The validated AG analytical method was then applied to 15 meats prepared from chicken, beef, and pork, and 6 culture media and additives used in cultured meat. No AGs were detected in any of the 15 meats distributed in Korea; however, streptomycin and dihydrostreptomycin were detected at levels ranging from 695.85 to 1152.71 mg/kg and 6.35 to 11.11 mg/kg, respectively, in the culture media additives. The LC-MS/MS method coupled with direct addition of IPRs to the vial can provide useful basic data for AG analysis and safety evaluation of meats as well as culture media and additives for cultured meats.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.151-169
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• 1993

The present study was carried out to investigate the best condition for nuclear-cytoplasm fusion and in vitro culture of nuclear transplanted embryos and to investigate the production of nuclear transplanted offsprings. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulses at output voltage of 1.0, 1.5 and 2.0 kV/cm for 100, 150 and $200{\mu}$ sec to induce cell fusion. 1. The culture of intact or zona cut 2-cell embryos in the medium supplemented with cytochalasin B($5{\mu}g/m{\ell}$) and colcemide($0.1{\mu}g/m{\ell}$)for 30 and 60 minutes did not affect the development to later stage. 2. The in vitro developmental rates of group A(a nucleus from one of the blastomeres was removed) and B(electrofusion of group A) were significantly lower than that of control group(p<0.01). 3. When nuclear transplanted embryos were submitted to electrofusion, the significantly higher fusion rates of 2-cell donor nuclei were achieved at the electric field strength of DC 1.5kV/cm for 100 and $150{\mu}$ sec, DC 2.0 kV/cm for $100{\sim}200{\mu}$ sec than DC 1.0 kV/cm for 100 and $150{\mu}$ sec(p<0.01). The significantly higher fusion rates of 4-cell donor nuclei were achieved at DC 2.0 kV/cm for 100 and $150{\mu}$ sec than DC 1.0kV/cm for $100{\sim}200{\mu}$ sec(p<0.01). These fusion rates in 8-cell donor nuclei were 88.7~99.3%. 4. The developmental potency to blastocyst in 2- and 4-cell donor nuclei was significantly higher in DC 1.0 and 2.0 kV/cm for $100{\sim}200{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group (p<0.01). The developmental potency to blastocyst in 8-cell donor nuclei was significantly higher in DC 2.0 kV/cm for $100{\mu}$ sec treated group than in DC 1.0 kV/cm for $100{\mu}$ sec treated group and DC 2.0 kV/cm for 150 and $200{\mu}$ sec treated group(p<001). 5. The developmental potency to blastocyst after nuclear transplantation was significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 6. The success rate of nuclear injection into enucleated 2-cell embryos was significantly higher in 2-cell donor nuclei than in 4- or 8-cell donor nuclei(p<0.01). 7. The culture time taken for the nuclear transplanted 2-cell embryos to blastocyst stage was significantly longer in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01). 8. There was no significant difference in the developmental potency of nuclear transplanted embryos within the concentration of EGF at 0 to 15 ng per $m{\ell}$ of BMOC-3 solution. 9. The production rates of offspring after transfer of nuclear transplanted embryos to recipient mouse were significantly higher in 2-cell donor nuclei than in 8-cell donor nuclei(p<0.01).