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Optimization of Compound K Production from Ginseng Extract by Enzymatic Bioconversion of Trichoderma reesei (Trichoderma reesei 유래 산업효소를 이용한 인삼추출물로부터 Compound K 생산 최적화)

  • Han, Gang;Lee, Nam-Keun;Lee, Yu-Ri;Jeong, Eun-Jeong;Jeong, Yong-Seob
    • The Korean Journal of Food And Nutrition
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    • v.25 no.3
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    • pp.570-578
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    • 2012
  • Compound K(ginsenoside M1) is one of saponin metabolites and has many benefits for human health. This study was to investigate Compound K produced from ginseng crude saponin extract with commercial cellulolytic complex enzyme(cellulase, ${\beta}$-glucanase, and hemicellulase) obtained from Trichoderma reesei. The effect factors(temperature, pH, ginseng crude saponin extract and enzyme concentration, and reaction time) on production of Compound K from ginseng crude saponin extract were determined by one factor at a time method. The selected major factor variables were ginseng crude saponin extract of 2%(w/v), enzyme of 7%(v/v), reaction time of 48 hr. Based on the effect factors, response surface method was proceeded to optimize the enzymatic bioconversion conditions for the desirable Compound K production under the fixed condition of pH 5.0 and $50^{\circ}C$. The optimal reaction condition from RSM was ginseng crude saponin extract of 2.38%, enzyme of 6.06%, and reaction time of 64.04 hr. The expected concentration of Compound K produced from that reaction was 840.77 mg/100 g. Production of Compound K was 1,017.93 mg/100 g and 862.31 mg/100 g, by flask and bench-scale bioreactor($2.5{\ell}$) system, respectively.

The comparison of IL-6, elastase and ${\alpha}1-PI$ expressions in human chronic periodontitis with type 2 diabetes mellitus (단순 만성 치주염 환자 및 2형 당뇨병환자의 만성치주염 치은조직에서 IL-6, elastase 및 ${\alpha}_1-PI$의 발현 양상 비교)

  • Park, Jae-Wan;Lee, Jae-Mok
    • Journal of Periodontal and Implant Science
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    • v.37 no.sup2
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    • pp.325-338
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    • 2007
  • The purposes of this study were to compare and quantify the expression of IL-6, e1stase and ${\alpha}_1-PI$ in the gingival tissues of patients with type 2 diabetes mellitus and healthy adults with chronic periodontitis. Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was devided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflammed gingiva from patients with chronic periodontitis. Group 3 (n=8) is inflammed gingiva from patients with chronic periodontitis associated with type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantification of IL-6, elastase and ${\alpha}_1-PI$ were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. 1. The expression levels of IL-6 showed increasing tendency in group 2 and 3, and It was highest in group 3. 2. The expression of elastase showed increasing tendency in group 2 and 3, and It was highest in group 3. 3. The expression of ${\alpha}_1-PI$ showed increasing tendency in group 3 compared to group 1. 4. The ${\alpha}_1-PI$/elastase ratio was decreased in group 2 and 3 compared to group 1, especially most decreased in group 3. 5. As IL-6 levels were increasing, elastase showed increasing tendency in group 3, and although IL-6 and elastase levels were increasing, ${\alpha}_1-PI$ level in group 3 showed slightly increasing pattern comparing to group 1. In conclusion, this study demonstrated that the expression levels of IL-6 and elastase will be inflammatory markers of periodontal inflammed tissue and DM. The ${\alpha}_1-PI$/elastase ratio also may be important measuring inflmmatory factors in the progression of periodontal inflammation associated to type 2DM.

NMR Spectroscopy and Mass Spectrometry of Phenylethanol Galactoside synthesized using Escherichia coli 𝛽-Galactosidase (대장균 베타-갈락토시데이즈를 이용하여 합성된 Phenylethanol Galactoside의 NMR Spectroscopy 및 Mass spectrometry)

  • Lee, Hyang-Yeol;Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.5
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    • pp.1323-1329
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    • 2020
  • To characterize the molecular structure of PhE-gal synthesized using Escherichia coli 𝛽-gal, NMR (1H- and 13C-) spectroscopy and mass spectrometry of PhE-gal were conducted. 1H NMR spectrum of PhE-gal showed multiple peaks corresponding to the galactosyl group, which is an evidence of galactosylation on 2-phenylethanol (PhE). Downfield proton peaks at 𝛿H 7.30~7.21 ppm showed the presence of aromatic protons of PhE as well as benzyl CH2 protons at 𝛿H 2.88 ppm. Up field proton peaks at 𝛿H 4.31 ppm, 4.07 ppm and multiple peaks from 𝛿H 3.86~3.38 ppm are indicative of galactocylation on PhE. 13C NMR spectrum revealed the presence of 12 carbons suggestive of PhE-gal. Among 12 carbon peaks from PhE-gal, the four peaks at 138.7, 129.0, 128.6 and 126.5 were assigned aromatic carbons in the phenyl ring. Three peaks at 129.0, 128.6 and 126.5 showed high intensities, indicating CH aromatic carbons. 13C NMR data of PhE-gal showed 6 monosaccharide peaks from galactose and 2 peaks from aliphatic chain of PhE, indicating that PhE-gal was galactosyl PhE. The mass value (sodium adduct ion of PhE-gal, m/z = 307.1181) from mass spectrometry analysis of PhE-gal, and 1H and 13C NMR spectral data were in good agreement with the expecting structure of PhE-gal. We are expecting that through future study it will eventually be able to develop a new additive with low cytotoxicity.

Daily Nutritional Intake and Serum Levels of Lipoprotein, Cholesterol and Protein -A Study of Buddhist Nuns- (한국여승(韓國女僧)의 영양섭취(營養攝取)와 혈청(血淸) Lipoprotein, Cholesterol 및 단백량(蛋白量)의 관계(關係))

  • Kim, Nan-Hee;Yoon, Jin-Sook;Choo, Young-Eun;Lee, Won-Jung
    • The Korean Journal of Physiology
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    • v.16 no.2
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    • pp.195-207
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    • 1982
  • To determine the influence of vegetarian diet on serum lipoprotein, cholesterol and protein levels, 45 young Buddhist nuns (age: $20{\sim}34$ years) and 29 female students(age: $20{\sim}22$ years) were examined. Daily caloric intakes were 1,945 Kcal for the Buddhist nuns and 1,815 Kcal for the students. The ratio of% calorie of carbohydrate: protein: fat from total calories in the Buddhist nuns was 84:11:5 and that in the students was 70:15:15. The Buddhist nuns had significantly higher carbohydrate intake but markedly lower lipid intakes than the students. Anthropometric measurement showed that the Buddhist nuns had significantly higher values of body weight, skin-fold thickness, body surface area and obesity index than the students. Both systolic and diastolic pressures of the Buddhist nuns and students were similar. Serum levels of total lipid, cholesterol and proteins in the Buddhist nuns were not different from those of the students. However, when comparing the levels of high density lipoprotein (HDL), very low density lipoprotein (VLDL) and low density lipoprotein (LDL) fractions, the Buddhist nuns had lower level of HDL but significantly higher LDL levels than the students. Furthermore, the Buddhist nuns had significantly lower levels of serum HDL-cholesterol but significantly higher LDL-cholesterol levels. There were significant correlations between LDL and LDL cholesterol (r=0.40), VLDL and VLDL-cholesterol(r=0.85), HDL and HDL-cholesterol(r=0.45), total serum lipid and total cholesterol (r=0.66) and total serum cholesterol and LDL(r=0.79). On the other hand, values of both serum total protein, and fractions of serum proteins were similar in the Buddhist nuns and students(ratio of albumin: ${\alpha}_{1}-:\;{\alpha}_{2}-:\;{\beta}-:\;{\gamma}-$globulins=55:3:10:13:19). Hematocrit and hemogloblin levels were similar in the Buddhist nuns and students. Above results suggest that vegetarian diets of the Buddhist nuns produced alterations in the metabolism of the lipoproteins and cholesterol.

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PET/CT SUV Ratios in an Anthropomorphic Torso Phantom (의인화몸통팬텀에서 PET/CT SUV 비율)

  • Yeon, Joon-Ho;Hong, Gun-Chul;Kang, Byung-Hyun;Sin, Ye-Ji;Oh, Uk-Jin;Yoon, Hye-Ran;Hong, Seong-Jong
    • Journal of the Korean Society of Radiology
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    • v.14 no.1
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    • pp.23-29
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    • 2020
  • The standard uptake values (SUVs) strongly depend on positron emission tomographs (PETs) and image reconstruction methods. Various image reconstruction algorithms in GE Discovery MIDR (DMIDR) and Discovery Ste (DSte) installed at Department of Nuclear Medicine, Seoul Samsung Medical Center were applied to measure the SUVs in an anthropomorphic torso phantom. The measured SUVs in the heart, liver, and background were compared to the actual SUVs. Applied image reconstruction algorithms were VPFX-S (TOF+PSF), QCFX-S-350 (Q.Clear+TOF+PSF), QCFX-S-50, VPHD-S (OSEM+PSF) for DMIDR, and VUE Point (OSEM) and FORE-FBP for DSte. To reduce the radiation exposure to radiation technologists, only the small amount of radiation source 18F-FDG was mixed with the distilled water: 2.28 MBq in the 52.5 ml heart, 20.3 MBq in the 1,290 ml liver and 45.7 MBq for the 9,590 ml in the background region. SUV values in the heart with the algorithms of VPFX-S, QCFX-S-350, QCFX-S-50, VPHD-S, VUE Point, and FOR-FBP were 27.1, 28.0, 27.1, 26.5, 8.0, and 7.4 with the expected SUV of 5.9, and in the background 4.2, 4.1, 4.2, 4.1, 1.1, and 1.2 with the expected SUV of 0.8, respectively. Although the SUVs in each region were different for the six reconstruction algorithms in two PET/CTs, the SUV ratios between heart and background were found to be relatively consistent; 6.5, 6.8, 6.5, 6.5, 7.3, and 6.2 for the six reconstruction algorithms with the expected ratio of 7.8, respectively. Mean SNRs (Signal to Noise Ratios) in the heart were 8.3, 12.8, 8.3, 8.4, 17.2, and 16.6, respectively. In conclusion, the performance of PETs may be checked by using with the SUV ratios between two regions and a relatively small amount of radioactivity.

Biological activity in hot water extract from fermented Coix lacryma-jobi L. var. mayuen Stapf. by Hericium erinaceum (Bull. : Fr.) (노루궁뎅이균사발효 율무 열수추출물의 유용성분 및 생리활성)

  • Yun, Kyeong-Won;Kim, Kyung-Je;Jin, Seong-Woo;Koh, Young-Woo;Im, Seung-Bin;Ha, Neul-I;Jeong, Hee-Gyeong;Jeong, Sang-Wook;Seo, Kyoung-Sun
    • Journal of Mushroom
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    • v.18 no.1
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    • pp.20-28
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    • 2020
  • This study was conducted to determine the antioxidant, nitrite scavenging, melanin tyrosinase inhibitory, and melanogenesis inhibitory activities of fermented Coix lacryma-jobi L. var. mayuen Stapf. by Hericium erinaceum (Bull.: Fr.) mycelial hot water extract (FCLHE). Additionally, we analyzed β-glucan and ergosterol contents in FCLHE and C. lacryma-jobi hot water extract (CLHE). The ergosterol and β-glucan contents in FCLHE were 740.2 mg% and 245.3 mg%, respectively, whereas these components were not detected in CLHE. FCLHE showed higher cell viability than CLHE. When B16F10 cells were treated with 500 ㎍/mL each of CLHE and FCLHE, the FCLHE treated cells produced 8.9 uM nitric oxide (NO), which was lower than that produced by CLHE treated cells (10.6 uM). The FCLHE treated cells showed significantly greater tyrosinase inhibition and melanin production at all tested concentrations than when compared to the CLHE treated group. Antioxidant parameters such as DPPH and ABTS radical scavenging activities were higher in FCLHE than in CLHE. These results suggest that FCLHE can be used as a raw material for functional foods, for food additives, and in the cosmetic industry.

Biodistribution and Scintigraphy of Iodine-131-Iododeoxyadenosine in Rats Bearing Breast Cancer (흰쥐에서 Iodine-131-Iododeoxyadenosine의 생체분포 및 유방암 영상화에 관한 연구)

  • Kim, Seon-Gu;Kim, Chang-Guhn;Lee, Kang-Mo;Kim, Hye-Won;Min Byung-Cheol;Choi, See-Sung;Lee, Jong-Deuk;Yang, David J.;Kim, E. Edmund;Lee, Hyun-Chul;Won Jong-Jin
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.4
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    • pp.374-381
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    • 1998
  • Purpose: I-131 labeled (2'-deoxy-2'-iodo-${\beta}$-D-arabinofuranosyl) adenine (IAD) may be involved in DNA synthesis during active proliferation of tumor cells. We conducted this study to find out the biodistribution of IAD and it's feasibility for scintigraphic tumor imaging. Materials and Methods: Tosyl acetyl-adenosine was dissolved in acetonitrile, and I-131-NaI was added and heated to synthesize IAD. Female Fisher 344 rats innoculated with breast tumor cells were injected with 0.27 MBq of IAD. Rats were sacrificed at 0.5, 1, 2, 4, 24h and the % of injected dose per gram of tissue (%ID/g) was determined. For scintigraphy, rats bearing breast cancer were administered with 1.11 MBq of IAD and imaging was performed after 2 and 24h. Then, rat body was fixed and microtomized slice was placed on radiographic film for autoradiography. Results: %ID/g of tumor was 0.74 (0.5h),0.73 (1h), 0.55 (2h), 0.38 (4h), and 0.05 (24h), respectively. At 1h after injection, %ID/g of tumor was higher than that of heart (0.34), liver (0.42), spleen (0.47), kidney (0.69), muscle (0.14), bone (0.33) and intestine (0.51). However, %ID/g of tumor was lower than blood (1.06), lung (0.77), and thyroid (177.71). At 4h, %ID/g of tumor in comparison with other tissue did not change. Tumor contrast expressed by tumor to blood ratio was 0.69 and tumor to muscle ratio was 5.11 at 1h. However, these ratios did not improve through 24h. On autoradiogram and scintigraphy at 2 and 24 hour, the tumor was well visualized. Conclusion: This results suggest that IAD may have a potential for tumor scintigraphy. However, further work is needed to improve localization in tumor tissue.

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Causative Organisms of Neonatal Sepsis (신생아 패혈증의 원인 병원체에 대한 조사)

  • Kim, Kyung-Ah;Shin, Son-Moon;Moon, Han-Ku;Park, Young-Hoon
    • Journal of Yeungnam Medical Science
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    • v.16 no.1
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    • pp.60-68
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    • 1999
  • A nationwide survey was conducted to investigate the annual occurrence rate of neonatal sepsis, maternal risk factors in neonatal sepsis, localized infection in neonates, causative organisms in nosocomial infection and the most common causative organism for neonatal sepsis in Korea. Clinical and bacteriological data wele collected from 37 neonatal units to perform retrospective review of the medical records of the newborn infants who were confirmed as having neonatal sepsis and whose blood culture was collected to isolate organisms for one year study period from January to December in 1997. 78,463 neonates were born at 37 hospital in 1997, and 20,869 neonates were admitted to the neonatal units, During this period, 772 episodes of neonatal sepsis were recorded in 517 neonates. The occurrence rate of neonatal sepsis was 0.73%(0~2.95%). Male to female ratio was 1.15:1, and 303 cases(42.1%) were born prematurely. The main pathogens of early onset of sepsis were S. aureus(20%), S. epidermidis(14.4%) and coagulase negative staphylococcus(14. 4%). Gram negative bacilli including Enterobacter spp (7.2%), E. coli(5.1%), Klepstella(4.5%), Pseudomonas(3.7%) and Enterobacter faectum(3.6%) accounted for 24.1% of sepsis. Group B beta-hemolytic streptococcus were isolated only in two cases. Common obstetric factors were PROM(21.1%), difficulty delivery(18.7%), fetal tachycardia(5.3%), chorioamnionitis(4.9%), and maternal fever(4.7%). The main pathogens of late-onset sepsis were S. aureus(22.3%), S. epidermidis(20.4%) and CONS(9.9%). There were 6 cases(1.0%) of Candida sepsis, Frequent focal infections accompanying sepsis were pneumonia(26.1%), urinary tract infection(10.5%), meningitis(8.2%), and arthritis(3.6%), S. epidermidis(22.0%) and s. aureus(21.7%) were also the most common pathogens in 373 nosocomial infection.

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Potential Reproductive Toxicity Study of p53 Expressing Adenoviral Vector in Mice (아데노바이러스 유전자치료벡터의 생식독성 연구)

  • Rhee, Gyu-Seek;Kwack, Seung-Jun;Kim, Soon-Sun;Lee, Rhee-Da;Seok, Ji-Hyun;Chae, Soo-Young;Chung, Soo-Youn;Kim, Seung-Hee;Lee, Seung-Hoon;Park, Kui-Lea
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.151-158
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    • 2007
  • The possibility of inadvertent introduction of therapeutic gene expressing viral vectors has raised safety concerns about germ-line infection. Particularly, for indications such as prostate cancer and ovarian cancer, the proximity of the point of viral administration to organs of the reproductive system raises concerns regarding inadvertent germ-line transmission of genes carried by the virus vector. To evaluate the safety of in vivo adenovirus mediated gene transfer, we explored the biodistribution, persistance and potential germ-line transmission of p53-expressing adenovirus (Ad-CMV-p53). Both male and female Balb/c mice were injected with $1{\times}10^9$ PFU of Ad-CMV-p53. The PCR analysis showed that there were detectable vector sequences in liver, kidney, spleen, seminal vesicle, epididymis, prostate, ovary, and uterus. The RT-PCR analysis for detecting inserted gene, p53 showed that Ad-CMV-p53 viral RNA were present in spleen, prostate and ovary. Direct injected male and female mice of adenovirus vector into testis and ovary were mated and their of offspring were evaluated for germ-line transmission of the adenoviral vector. The PCR and RT-PCR analysis showed no evidence of germline transmission, although vector sequences were detected in DNA extracted from gonadal tissues. Real-time PCR result confirmed a significant decrease of adenovirus in gonad tissues 1 week after injection. We have also analysed the cell specific localization of viral DNA in gonad tissues by using in-situ PCR. Positive signals were detected in interstitial tissue but not in seminiferous tubule in sperm. In the case of ovary, adenovirus signal were localized to the stromal tissue, but no follicular signals were observed. Together, these data provide strong evidence that the risk of the Inadvertent germ-line transmission of vector sequences following intraperitoneal or direct injection into genito-urinary system of adenovirus is extremely low.

Effects of Manipulation Conditions on Development of Nuclear Transplant Bovine Embryos Derived from In Vitro Matured Oocytes (미세조작조건이 소 핵이식배의 발달에 미치는 영향)

  • 최상용;노규진;공일근;송상현;조성근;박준규;이효종;박충생
    • Korean Journal of Animal Reproduction
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    • v.21 no.3
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    • pp.293-302
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    • 1997
  • Follicular oocytes of Grade I and II were collected from 2~6 mm ovarian follicles and matured in vitro (IVM) for 24 hrs in TCM-199 su, pp.emented with 35$\mu\textrm{g}$/ml FSH, 10$\mu\textrm{g}$/ml LH, and 1$\mu\textrm{g}$/ml estradiol-17$\beta$ at 39$^{\circ}C$ under 5% CO2 in air. They were fretilized in vitro (IVF) by epididymal spermatozoa capacitated with heparin for 12 hrs. The zygotes were then co-cultured in vitro with bovine oviducted epithelial cells (BOEC) for 7 to 9 days. The optimal time for IVM, the successful enucleation of IVM oocytes by micromanipulation at different oocyte ages after IVM, and the ideal culture system for IVM for effective IVF and in vitro development of IVM-IVF embryos was examined for in vitro production of nuclear recipient oocytes and nuclear donor embryos. To improve the efficiency of nuclear transplantation (NT) of IVF embryo into IVM follicular oocytes, this study evaluated the optimal electric condition and oocytes age for activation of IVM oocytes and in vitro development of NT embryos. In vitro development of NT embryos with preactivation or non-preactivation in enucleation oocytes, cell number of IVN-IVF embryos, and NT embryos wre also examined. The results obtained were as follows; 1. The most suitable enucleation time was at 24 hpm (83.3%) rather than that of 28 hpm(69.6%) and 32 hpm(50.0%). 2. There was no difference among the fusion rates of NT embryos at the voltages of 0.75, 1.0 and 1.5 kV/cm, but the in vitro development rates to morule and blastocyst were significantly (P<0.05) higher at the voltage of 0.75(12.5%) and 1.0kV/cm (12.6%) compared to 1.5kV/cm(0%). 3. No significant difference in activation rates were seen in NT embryos stimulated for 30, 60 and 120 $\mu$sec (71.7, 85.2 and 71.9%, respectively), but the in vitro development rates to morulae and blastocyst were significantly (P<0.05) higher in the oocytes stimulated for 30 $\mu$sec (11.6%) and 60 $\mu$sec(10.7%) than 120 $\mu$sec(0.0%). 4. The fusion rates (71.0 and 87.3%) and the in vitro development rates (9.1 and 12.7%) to morula and blastocyst were seen in the NT embryos stimulated at 28 and 32 hpm under the condition of 1.0 kV/ml, 60 $\mu$sec. However, at 24 hpm the fusion rates were 64.8% and the in vitro development to morula and blastocyst were not seen. 5. The fusion rates between the 8~12, 13~17 and 18~22-cell stage of IVM-IVF embryos were not significantly different. The in vitro development rates of the fused embryos to morula and blastocyst which were received from a blastomere of 8~12, 13~17 and 18~22-cell stages of IVM-IVF embryos were 14.9, 8.3 and 6.5%, respectively. 6. The in vitro development rate of the enucleated recipient oocytes with preactivation (24.2%) to morula and blastocyst was significantly (P<0.05) higher than that of non-preactivation (12.8%). 7. The cell numbers of NT blastocyst and IVM-IVF blastocyst cultured during 7~9 days were 63$\pm$11 and 119$\pm$23, and then their the mean cell cycle number were 5.98 and 6.89, respectively.

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