• Title/Summary/Keyword: 미세주입

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Studies on the Efficient Embryo Transfer Methods using Inbred Embryos in Generation of Transgenic Mice (Inbred 마우스 수정란을 이용한 형질전환마우스 생산에 있어서의 효과적인 수정란 이식 방법에 관한 연구)

  • Wang A. G;Kim, S. U.;Y. H. Han;Kim, S. K.;D. Y. Yu
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.69-75
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    • 2003
  • The objective of this study is to improve the efficiency of embryo transfer in generation of transgenic mice using inbred mouse (C57BL/6J) embryos. The embryos of C57BL/6J and BCF1 mice were superovulated by the standard protocol. One-cell stage of embryos were microinjected and the resulted one-or two-cell were transferred into one-or two-side oviducts of foster mother, respectively. When one-cell stage of embryos were transferred into one-side oviduct of 0.75 d.p.c. foster mother, the results were not ideal because of showing pregnancy ratios of 68.8$\pm$7.83% for C57BL/6J and 48.3$\pm$14.22% for BCF1, and development ratios of pups vs transferred embryos of 11.9$\pm$5.51% for C57BL/6J and 10.5$\pm$8.03% fur BCF1 . However, when two-cell stage of embryos were transferred into two-side oviducts of 0.5 d.p.c. foster mother, we got better results of 94.4$\pm$9.64% and 100$\pm$0% pregnancy ratio, and 22.1$\pm$0.4% and 21.8$\pm$0.38% development ratio for C57BL/6J and BCF1, respectively. Therefore, transferring two-cell stage of C57BL/6J embryos into two-side oviducts of 0.5 d.p.c. foster mother may be competitive to the result in BCF1 embryos. Even if there are a lot of other factors affecting these results, we conclude from these experiments that transfer of two-cell embryos into two-side oviducts of 0.5 d.p.c. foster mother is a more efficient and safe method than others in generating transgenic mice using inbred mice embryos.

Effects of Indoor Greening Method on Temperature, Relative Humidity and Particulate Matter Concentration (실내녹화 방법이 온·습도 및 미세먼지 농도에 미치는 영향)

  • Kwon, Kei-Jung;Park, Bong-Ju
    • Journal of the Korean Institute of Landscape Architecture
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    • v.45 no.4
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    • pp.1-10
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    • 2017
  • This study investigated indoor temperature and humidity control and PM1 and PM10 mitigation effects of a single green wall (Case 1), two green walls (Case 2), and two green walls plus a waterfall (Case 3) in comparison with a control without either a green wall or waterfall. Experiments were conducted in the office of Chungbuk National University from August to September, 2015. Experiments were carried out sequentially in the order of control, Case 1, Case 2, and Case 3. Data collected from August 17 to August 20, 2015 (Experiment 1), and from August 31 to September 3, 2015 (Experiment 2), when outdoor temperature was relatively constant, were analyzed. Plant volume ratios by indoor landscaping of the control, Case 1, Case 2 and Case 3 were 0.0, 0.6, 1.2, and 1.4%, respectively. Compared to the control, average temperatures of Case 1, Case 2 and Case 3 were decreased by 0.3~0.7, 0.7~0.9 and $1.0^{\circ}C$, respectively, and relative humidity was increased by 1.8~8.7, 9.2~14.6 and 14.8~21.9%, respectively. Three hundred minutes after exposure to mosquito repellent incense particles, the ratio of the remaining PM1 of the control, Case 1, Case 2 and Case 3 were 25.0, 22.0%, 21.2%, 17.3%, respectively, in Experiment 1 and 42.3, 28.9, 23.1, and 30.9%, respectively, in Experiment 2. As indoor greening increased the effect of indoor temperature, PM1 and PM10 mitigation were greater, and temperature and humidity were lower. The greater the relative humidity was, the faster PM1 and PM10 mitigation tended to be.

Microassisted Fertilization of Human Oocytes with Intracytoplasmic Sperm Injection in IVF-ET Patients with History of Failure in Fertilization or Extremely Low Fertilization Rate in Previous Cycles (미수정 및 저수정율의 기왕력을 지닌 체외수정시술 환자에서의 난자 세포질내 정자 주입술을 이용한 미세보조 수정술에 관한 연구)

  • Moon, Shin-Yong;Kim, Seok-Hyun;Chae, Hee-Dong;Kim, Kwang-Rye;Lee, Jae-Hoon;Kim, Hee-Sun;Ryu, Buom-Yong;Oh, Sun-Kyung;Suh, Chang-Suk;Choi, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.24 no.1
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    • pp.83-93
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    • 1997
  • Although IVF-ET is widely applied in the treatment of couples with male factor infertility, it may fail in many infertile couples with normal semen parameters, and certain couples cannot be accepted for standard IVF-ET due to unfertilization or extremely low fertilization rate of oocytes. Recently, several procedures of microassisted fertilization (MAF) using micromanipulation have been introduced, and pregnancies and births have been obtained after partial zona dissection (PZD), subzonal insertion (SUZI), and intracytoplasmic sperm injection (ICSI). This clinical study was performed to develop and establish ICSI as an effective procedure of MAF in infertile couples who could not undergo standard IVF-ET repetitively because of failure in fertilization or extremely low fertilization rate of oocytes with the conventional fertilization technique in the previous IVF-ET cycles. From March, 1995 to May, 1996, 27 cycles of IVF-ET with ICSI in 19 infertile patients were included in study group, and the outcomes of ICSI were analyzed according to fertilization rate, cumulative embryo score (CES), and pregnancy rate. The number of oocytes retrieved after controlled ovarian hyperstimulation (COH) was $10.50{\pm}6.13$ in 30 previous cycles, and $10.57{\pm}5.53$ in 27 ICSI cycles. In ICSI cycles, the number of oocytes optimal for ICSI procedure was $7.89{\pm}4.30$, and the fertilization rate of $67.9{\pm}20.2%$ could be obtained after ICSI. The number of embryos transferred was $1.43{\pm}2.40$ in previous cycles, and $4.36{\pm}1.77$ with the mean CES of $41.8{\pm}27.4$ in ICSI cycles. In ICSI cycles, the overall pregnancy rate was 29.6% (8/27) per cycle and 42.1% (8/19) per patient with the clinical pregnancy rate of 22.2% (6/27) per cycle and 31.6% (6/19) per patient. In conclusion, MAF of human oocytes with ICSI is a promising fertilization method for IVF-ET patients, especially with the past history of failure in fertilization or low fertilization rate of oocytes in the previous IVF-ET cycles, and ICSI using micromanipulation procedures applied to human oocytes will provide a range of novel techniques which may dramatically improve the pregnancy rate in IVF-ET program and contribute much to effective management of infertile couples.

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Sorghum Extract Lowers Lymphatic Absorption of Trans Fat and Cholesterol in Rats (흰쥐에서 수수추출물이 트랜스지방산이 함유된 지방과 콜레스테롤의 흡수에 미치는 영향)

  • Kim, Juyeon;Noh, Sang Kyu;Woo, Koan-Sik;Seo, Myung-Chul
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.6
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    • pp.783-788
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    • 2016
  • Excessive intake of trans fats is a risk factor for cardiovascular diseases. Previously, we showed that green tea extract lowers intestinal absorption of cholesterol and elaidic acid in rats. This study was conducted to investigate a possible role for sorghum extract on lymphatic absorption of trans fat and cholesterol in rats. Adult male rats with lymph cannulae were infused at a rate of 3.0 mL/h for 8 h via a duodenal catheter with a lipid emulsion containing $146.4{\mu}mol$ of trielaidin, $36.8{\mu}mol$ of trilinoelaidin, $452.0{\mu}mol$ of triolein, $1.0{\mu}Ci$ cholesterol labeled with $^{14}C$ ($^{14}C-cholesterol$), $20.7{\mu}mol$ of cholesterol, and $396.0{\mu}mol$ of Na-taurocholate without or with 100.0 mg of sorghum extract in phosphate-buffered saline buffer (pH 6.4). Lymph was collected hourly for 8 h. No significant difference was noted in lymph flow. However, the lymphatic absorption of elaidic acid and linoelaidic acid for 8 h was significantly lower in rats infused with sorghum extract than in those infused with no sorghum extract. Further, lymphatic absorption of $^{14}C-cholesterol$ was reduced by sorghum extract, which was observed previously. These data indicate that sorghum extract has an inhibitory effect on the intestinal absorption of trans fat and cholesterol. The mechanism(s) by which sorghum extract lowers intestinal absorption of trans fat warrants further study.

The Change of Alveolar-capillary Barrier by Germanium in Acute Lung Injury Induced by Lipopolysaccharide (LPS에 의한 급성 폐손상에서 게르마늄에 의한 폐포-모세혈관 장벽의 변화)

  • Lee, Yoon-Jeong;Cho, Hyun-Gug;Sin, Gun-Ho;Jeune, Kyung-Hee
    • Applied Microscopy
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    • v.39 no.1
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    • pp.27-40
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    • 2009
  • Acute respiratory distress syndrome (ARDS), also known as an acute inflammatory lung disease is developed by various factors that is originated from the destruction of alveolar-capillary barrier, and neutrophils plays an important role in the destruction. The study intended to confirm, the anti-inflammatory effect of germanium, whether a lung injury has been mitigated with the reduction of injury in alveolar-capillary barrier resulting from inhibition of neutrophils migration in lung tissue. Test groups were divided in saline administered CON, 5 hours of endotoxin administered LPS and 5 hours of endotoxin administered Ge+LPS following 1 hours of pre-processed germanium. $100{\mu}g$ endotoxin was melted in 0.5 mL saline and sprayed into airway and 26 mg germanium per 100 g weight was administered into abdominal cavity. The endotoxin group which induced an acute lung injury with administered endotoxin showed dramatic increase of pulmonary edema (p<0.001), protein contents in bronchoalveolar lavage fluid, BALF (p<0.05) and neutrophils of infiltration in BALF (p<0.001) comparing with a control group, while a pre-treated germanium group showed significant decrease in all categories comparing to the endotoxin administerd group. In the result of a microscopic observation, the structure of alveolar-capillary barrier which is constructed with basal lamina, alveolar type I cells and endothelial cell were preserved of the pre-treated germanium group relatively well compare to the endotoxin administered group. And the construction of lamellar body, microvilli and basal lamina of alveolar type II cells were also preserved relatively well. Hence, germanium activates as an anti-Inflammatory mediator in other words, it interfered neutrophils migration into lung tissue, thereby reduced injury of alveolar-capillary barrier from toxic substances of activated neutrophils. Consequently, the study has determined that the acute lung injury induced by endotoxin has been decreased by the pre-treated germanium.

The Increase of Apoptotic Neutrophils and Phagocytic Macrophage by Germanium in Acute Lung Injury Induced by Lipopolysaccharide (LPS에 의한 급성 폐손상에서 게르마늄에 의한 호중구 세포사와 큰포식세포의 포식능 증가)

  • Lee, Yoon-Jeong;Cho, Hyun-Gug;Jeune, Kyung-Hee
    • Applied Microscopy
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    • v.38 no.4
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    • pp.293-306
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    • 2008
  • The essential factor of acute respiratory distress syndrome (ARDS), an acute lung injury accompanied commonly by sepsis syndrome is accumulation of neutrophils in lung tissue. The study attempted to confirm whether a lung injury would be decreased with the anti-inflammatory effect of germanium by the treated germanium prior to the development of ARDS and whether nitric oxide influence in suppressing a lung injury. Test groups were divided in the following structure for experiment; CON that has been administered with sodium chloride to airway, LPS administered with endotoxin for 5 hours in the same amount and 5 hours of endotoxin administered Ge+LPS following 1 hours of pre-treated germanium. The result of a test using experimental animals, infilteration of neutrophils (p<0.001) in bronchoalveolar lavage fluid (BALF) was significantly decreased, the structure of lung tissue was preserved relatively well, and much neutrophils with distinct positive were observed on tunel staining which showed increase of apoptotic neutrophils in the pre-treated germanium group compare to the endotoxin administrated group. In observation of ultrastructural changes of cell in BALF, phagocytic alveolar macrophage was increased in alveolar space, the nucleus of most engulfed neutrophils were condensed, and some apoptosis neutrophils appears to be DNA fragmentation and effacement of cellular organelles were found in intercellular matrix in the pre-treated germanium group. However, the nitric oxide showed increase in all the groups excluding CON, and the nitric oxide effect such as degranulation diminishing of mast cells and apoptosis increase of neutrophils in the pre-treated group only. The situation appears that there was change in internal environment of the experimental animal by the pre-treated germanium before the nitric oxide is produced and the anti-inflammatory effect activated the pre-processed germanium by nitric oxide which activated following the change. Therefore, the nitric oxide created from macrophage in accordance with the pre-treated germanium appears to influence in alleviating a lung injury. Accordingly, acute lung injury is alleviated by the anti-inflammatory effect of germanium such as inhibition of neutrophils migration, induction of neutrophil apoptosis and increase of phagocytic function of phagocyte, and the nitric oxide produced from activated macrophage by germanium would influence in suppressing a lung injury.

Microstructure of ZnO Thin Film on Nano-Scale Diamond Powder Using ALD (나노급 다이아몬드 파우더에 ALD로 제조된 ZnO 박막 연구)

  • Park, S.J.;Song, S.O.
    • Journal of the Korean Vacuum Society
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    • v.17 no.6
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    • pp.538-543
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    • 2008
  • Recently a nano-scale diamond is possible to manufacture forms of powder(below 100 nm) by new processing of explosion or deposition method. Using a sintering of nano-scale diamond is possible to manufacture of grinding tools. We have need of a processing development of coated uniformly inorganic to prevent an abnormal grain growth of nano-crystal and bonding obstacle caused by sintering process. This paper, in order to improve the sintering property of nano-scale diamond, we coated ZnO thin films(thickness: $20{\sim}30\;nm$) in a vacuum by ALD(atomic layer deposition) Economically, in order to deposit ZnO all over the surface of nano-scale diamond powder, we used a new modified fluidized bed processing replaced mechanical vibration effect or fluidized bed reactor which utilized diamond floating owing to pressure of pulse(or purge) processing after inserted diamond powders in quartz tube(L: 20 mm) then closed quartz tube by porosity glass filter. We deposited ZnO thin films by ALD in closed both sides of quartz tube by porosity glass filter by ALD(precursor: DEZn($C_4H_{10}Zn$), reaction gas: $H_2O$) at $10^{\circ}C$(in canister). Processing procedure and injection time of reaction materials set up DEZn pulse-0.1 sec, DEZn purge-20 sec, $H_2O$ pulse-0.1 sec, $H_2O$ purge-40 sec and we put in operation repetitive 100 cycles(1 cycle is 4 steps) We confirmed microstructure of diamond powder and diamond powder doped ZnO thin film by TEM(transmission electron microscope) Through TEM analysis, we confirmed that diamond powder diameter was some $70{\sim}120\;nm$ and shape was tetragonal, hexagonal, etc before ALD. We confirmed that diameter of diamond powders doped ZnO thin film was some $70{\sim}120\;nm$ and uniform ZnO(thickness: $20{\sim}30\;nm$) thin film was successfully deposited on diamond powder surface according to brightness difference between diamond powder and ZnO.

In Vitro Development of Bovine Nuclear Transfer Embryos Reconstructed with Fetal Fibroblasts (태아 섬유아세포로 재구성된 핵치환 소 수정란의 체외발달)

  • Koo, D.B.;Choi, Y.H.;Park, J.S.;Kim, H.N.;Kang, Y.K.;Lee, C.S.;Han, Y.M.;Park, H.D.;Lee, K.K.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.407-417
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    • 2000
  • The present study was to examine effects of various electrical stimulus treatments used for electro-fusion on the preimplantation development of bovine nuclear transfer (NT) embryos with fetal fibroblast cells. Fetal fibroblast cells were isolated from one fetus at day 45 of gestation in Holstein cow, and passaged 3 to 4 times before being transferred into enucleated oocytes. Single fibroblast cells were individually placed into the perivitelline space of enucleated oocytes by using a micromanipulator. At first, the fusion and developmental rates of reconstructed oocytes were compared between different electric stimulation conditions. When fusion of the reconstructed oocyte was induced by different electric pulse periods (15, 30 and 45 $\mu$sec) at a DC pulse of 1.8 kV/cm, 15 (45.5%, 120/264) or 30 $\mu$ sec group (43.9%, 106/241) showed a higher fusion rate than 45 $\mu$sec group (23.2%, 58/250, P<0.05). However, no difference was detected in the development rate of the fused oocytes to blastocysts between groups. Next experiment was to examine the effects of different electrical field strengths (1.5, 1.8 and 2.1 kV/cm) for 15 $\mu$sec at electrofusion on in vitro development of the NT embryos. As results, there was no difference in the fusion and developmental rates of the NT embryos between electrical strength (P>0.05). Finally, developmental competence of bovine NT embryos with somatic cells was compared with IVF-derived embryos. Of enucleated oocytes fused with fibroblast cells, 27.4% (75/274) developed to the blastocyst stage, which is similar to that (24.5%, 58/237) of IVF-derived embryos. However, mean nuclei number of NT blastocysts was smaller than that of IVF-derived blastocysts. Thus, we have established an optimal condition (1.8 kV/cm, 15 $\mu$sec) for electric fusion of bovine NT oocytes with somatic cells. The present study indicates that bovine reconstructed embryos with somatic cells normally develop to blastocyst stage in vitro, although having smaller nuclei numbers of blastocysts as compared to IVF-derived embryos.

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Factors Affecting Pregnancy Rates on Transfer of Pronuclear Microinjected Embryos in Korean Black Goats (전핵 미세 주입법으로 생산된 한국흑염소 수정란의 이식 조건이 수태율에 미치는 영향)

  • Choi, Y.S.;Shin, H.G.;Jang, S.K.;Yang, H.S.;Lee, O.K.;Lee, D.S.;Cho, J.K.;Shin, S.T.
    • Journal of Embryo Transfer
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    • v.22 no.1
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    • pp.53-61
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    • 2007
  • This study was investigated factors affecting the pregnancy rates after transfer of pronuclear microinjected embryos for the production of transgenic Korean black goats. Embryo transfer was carried out in 343 recipient Korean black goats from September 1999 to June 2000. Estrus was induced by the insertion of intravaginal progesterone devices $CIDR^(R)$ for 2 weeks. A single injection of 400 IU equine chorionic gonadotropin was administered at 48h before $CIDR^(R)$ removal to increase the proportion of does cycling and ovulation rate. Good quality embryos were prepared by microinjection of DNA into the pronuclei of fertilized goat oocyte and cultured in vitro. Pronuclear microinjected $1{\sim}8$ cell stage embryos were surgically transferred into the oviducts of the recipient at day 4 or 5 following $CIDR^(R)$ removal, and morula to blastocyst stage embryos were surgically transferred into uterus at day 9. Pregnancy was diagnosed by transrectal ultrasound scanning at $20{\sim}30d$ and 8 weeks following embryo transfer. The pregnancy rate was affected by several factors, such as estrus induction, the number of previous transfer, transfer site, stage of CL (corpus luteum), the number of recipient CL, stage of embryos and the number of transferred embryo. The pregnancy rate was significantly higher in recipients that came into estrus naturally than recipients that induced to come into estrus with $CIDR^(R)$(59.1% vs. 36.8%; P<0.05). The pregnancy rate was higher when the embryos were transferred into the left oviduct than transferred into the right oviduct (42.9% vs. 35.3%; P<0.05). The pregnancy rate of recipients with $CH_1$ (early) stage corpus hemorrhagicum in ovary was hi틴or than recipient with $CH_3$ (late) stage hemorrhagicum (47.5% vs. 17.9%; P<0.01). Higher pregnancy rates were obtained by transfer of 1-cell stage embryos into oviduct while late blastocysts (51.6% vs. 66.7%; P<0.01) into uterus. The pregnancy rates when 3 embryos were transferred to recipients were significantly higher than when 2 embryos we.e transferred (47.6% vs. 27.0%; P<0.05). Although there were no significant difference among the group, adhesion of reproductive organs, uterine size, ovulation rate of recipients, presence of large follicle and difficulty of transfer affected pregnancy rate of recipient. Higher pregnancy rates were obtained in the recipients with $8{\sim}15m$ diameter uterine horn as compared to the recipients with <5m diameter or >20mm diameter uterine hem (38.9%, 20% vs. 18.2%), in the recipients with large follicle in the ovulated ovary ipsilaterally (53.6% vs. 37.1%) and in the transfer which was carried out easily (39.2% vs. 27.8%, 0%). In conclusion, the high rate of pregnancy was achieved following transfer of pronuclear microinjected embryos when three or four 1-cell stage embryos were transferred into oviduct with $CH_1$ stage corpus hemorrhagicum in the ovary of recipient which came into estrus naturally.