• Clinical and Experimental Reproductive Medicine
• /
• v.27 no.1
• /
• pp.31-37
• /
• 2000

This study was designed to identify the ultrastructural changes of mouse endometrium during peri-implantation period and obtain the fundamental information for the establishment of 3-dimensional culture system of mouse endometrial cells in vitro. The used female ICR mice ($6{\sim}8$ wks) were conducted on pregnant. The biopsies were obtained from whole uterus at cycle day 1 (D1) and day 5 (D5) after hCG injection and mating. The biopsies materials were fixed 2.5% glutaraldehyde and 1% osmium tetroxide. Subsequently, for observation using light and transmission electron microscopy (LM and TEM), they were dehydrated and embedded in Epon and the embedded biopsies were sectioned and stained. For scanning electron microscopy (SEM), the fixed specimens were dehydrated, dried and coated with gold. 1) For LM, the biopsied materials at D5 (late secretory phase) were appeared the extended stromal layer by increased connective tissues and the fully developed endometrial glands and vessels compared with D1 (early secretory phase). 2) For TEM, the mouse endometrium was consisted of 3-layers, a simple polarized columnar epithelial cells, basement membrane and stromal cells. At D5, the distribution of microvilli, endoplasmic reticulum, Golgi body, lipid and glycogen deposits, secretory granules and surface area of basement membrane were increased. 3) For SEM, the degree of folding and microvilli of surface of mouse epithelial cells was became more and more according to the process of secretory phase, and at D5, implantation time of mouse, the appearance of pinopodes as a specific marker of uterine receptivity was found. The uterine pinopodes of mouse were found in narrow sites at the luminal surface, irregularity and appeared the different stages in the same sample. Therefore, these results indicated that the mouse endometrium was experienced dramatic morphological changes during peri-implantation period.

• Journal of Embryo Transfer
• /
• v.12 no.2
• /
• pp.133-139
• /
• 1997

Large scale production of cloned embryos requires the technology of multiple generational nuclear transfer(NT) by using NT embryos itself as the subsequent donor nuclei. In this work we investigated comparatively the effects of enucleated oocytes treated with ionomycin and 6-DMAP on the electrofusion rate and in vitro developmental potential in the first and second NT embryos. The embryos of 16-cell stage were collected from the mated does by flushing oviducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FCS) at 47 hours after hCG injection. The recipient cytoplasms were obtained by removing the nucleus and the first polar body from the oocytes collected at 15 hours after hCG injection. The enucleated oocytes were pre-activated by 5 min incubation in 5$\mu$M ionomycin and 2 hours incubation in 2 mM 6-DMAP at 19~20 hours post-hCG before microinjection. In the first and second generation NT, the unsynchronized 16-cell stage embryos were used as nuclear donor. The separated donor blastomeres were injected into the enucleated activated recipient oocytes by micromanipulation and were electrofused by electrical stimulation of single pulse for 60 $\mu$sec at 1.25kV/cm in $Ca^2$+, $Mg^2$+ - free 0.28 M mannitol solution. In the non-preactivation group, the electrofusion and electrical stimulation was given 3 pulses for 60 $\mu$sec at 1.25 kV/cm in 100$\mu$M $Ca^2$+, $Mg^2$+ 0.28 M mannitol solution. The fused oocytes were co-cultured with a monolayer of rabbit oviductal epithelial cells in TCM-199 solution containing 10% FCS for 120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The results obtained were summarized as follows: 1. In the first generational NT embryos, the electrofusion rate of preactivated and non-activated oocytes(80.4 and 87.8%) was not significantly different, but in the second generational NT embryos, the electrofusion rate was significantly(P<0.05) higher in the non-activated oocytes(85.7%) than in the preactivated oocytes(70.1%). 2) In the first and second generational NT embryos, the developmental potential to biastocyst stage was significantly(P<0.05) higher in the preactivated oocytes(39.3 and35.7%) than in the non-preactivated oocytes(16.0 and 13.3%). No significant difference in the developmental potential was shown between the first and second generational NT embryos derived from the preactivated oocytes. In conclusion, it may be efficient to use the oocytes preactivated with ionomycin and 6-DMAP for the multiple production of cloned embryos by recycling nuclear transfer.

• School Mathematics
• /
• v.10 no.1
• /
• pp.79-103
• /
• 2008

The purpose of this study is for developing and using project tasks which can be used to connect schooling and home education for the other days of 5days-schooling; to promote students' mathematical ability and to let students have positive cognitions toward mathematics and self-controled learning attitudes. For this study, two classes of 4th graders(56 students) were sampled from a school in D city. Half of them were assigned to the experiment group(EG) and the other to comparison group(CG). In the experiment group, students completed 16 project tasks and we investigated whether there is an effect on students' academic achievement and mathematical disposition. Two kinds of test instruments, pre-test and post-test were used. The pre-test scores guaranteed that both groups were homogeneous. Post-test scores were used to identify three effects and the post-test scores were analyzed by t-test. The result of this study is as follows: (1) There was significant difference between EG and CG in academic achievement (p=0.010). (2) There was significant difference between EG and CG in mathematical disposition(p=0.007). (3) There was no significant difference between EG's pre-test and post-test in mathematical disposition at the 5% significant level. But the average score of mathematical disposition improved from 3.3333 to 3.5375. Mathematical disposition was composed of 6 factors. One of them was mathematical value and there was significant difference between EG's pre-test and post-test at the 5% significance level(p=0.030). But other factors were not. The average scores of mathematical reflection improved a little. So we can say that the activities with project tasks brought an positive influence on students' mathematical disposition.

• Investigative Magnetic Resonance Imaging
• /
• v.25 no.3
• /
• pp.141-155
• /
• 2021

Purpose: To develop a 3D magnetic resonance fingerprinting (MRF) method for application in high resolution knee cartilage PD, T₁, T₂ mapping. Materials and Methods: A novel 3D acquisition trajectory with golden-angle rotating radial in k_xy direction and interleaved echo planar imaging (EPI) acquisition in the k_z direction was implemented in the MRF framework. A centric order was applied to the interleaved EPI acquisition to reduce Nyquist ghosting artifact due to field inhomogeneity. For the reconstruction, singular value decomposition (SVD) compression method was used to accelerate reconstruction time and conjugate gradient sensitivity-encoding (CG-SENSE) was performed to overcome low SNR of the high resolution data. Phantom experiments were performed to verify the proposed method. In vivo experiments were performed on 6 healthy volunteers and 2 early osteoarthritis (OA) patients. Results: In the phantom experiments, the T₁ and T₂ values of the proposed method were in good agreement with the spin-echo references. The results from the in vivo scans showed high quality proton density (PD), T₁, T₂ map with EPI echo train length (N_ETL = 4), acceleration factor in through plane (R_z = 5), and number of radial spokes (N_spk = 4). In patients, high T₂ values (50-60 ms) were seen in all transverse, sagittal, and coronal views and the damaged cartilage regions were in agreement with the hyper-intensity regions shown on conventional turbo spin-echo (TSE) images. Conclusion: The proposed 3D MRF method can acquire high resolution (0.5 mm³) quantitative maps in practical scan time (~ 7 min and 10 sec) with full coverage of the knee (FOV: 160 × 160 × 120 mm³).

• KIEAE Journal
• /
• v.13 no.1
• /
• pp.109-123
• /
• 2013

The study is to probe a technical alternative to enhance the reliability and accuracy of the results of various landscape simulations. This study to present technical criteria that are necessary in each stage of target site analysis, picture taking, and computer synthesis and, through these to present supplementary plans to enhance the reliability and accuracy of landscape simulations. In order to derive more practical and empirical results in terms of the reliability of the results of landscape simulations, examples that actually passed landscape review were selected. With regard to study process, an analysis was made first to analyze the landscape report data of designs that passed the review to analyze their characteristics, to be followed by an integrated analysis of problems that were revealed in various landscape simulations. Important factors that affect directly the work of landscape simulations such as the specification of camera lenses that were used in picture taking, distance, and angle. Design the work was carried out using Auto CAD, 3DS Max, and Photoshop program in the same way as in actual design. For verification of their accuracy and reliability, the results were entrusted to experts who have implemented similar jobs. To seduce differences from those too landscape simulations that conduct trial experiment of the virtual space that are to be created in the future with accurate numerical values.

• Journal of Embryo Transfer
• /
• v.11 no.3
• /
• pp.201-210
• /
• 1996

This experiment was carried out to evaluate the effect of early mouse embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells (BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated cultured in D-PBS /15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation in the embryos were examined under the fllowing treatments; 1) TCM 199 added 15% HCS, 2) Ham's F-10 added 15% HCS, 3) MediCult IVF medium, 4) TCM 199 added 15% HCS + BOEC, 5) TCM 199 added 15% HCS + POEC, 6) Ham's F40 added 15% HCS + BOEC, 7) Ham's F-10 added 15% HCS + POEC,8) MediCult IVF medium + BOEC, 9) MediCult IVF medium + POEC. For a comparative study of in vitro development for 96 hours after hCG injection, were cultured with oviductal epithelial cell and media only. The obtained results were 2-cell embryos developed to the blastocyst stage in TCM 199, Ham's F-10 and MediCult IVF medium at the rates of 84.4,83.2 and 81.6%. respectively. The higher developmental rates(91~97%) of blastocyst formation was appeared when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal epithelial cells but significant difference in co-culture system in comparison between media only system and co-cultures. In conclusions, oviductal epithelial cells, BOEC and POEC, when co-culture with mouse early embryos improved the rates of development, blastocyst and hatching. Therefore, it is suggested that co-culture system using oviductal epithelial cells improve early embryonic developtnent in mouse.

• Clinical and Experimental Reproductive Medicine
• /
• v.26 no.2
• /
• pp.179-183
• /
• 1999

This study was performed to determine the significance of a baseline ovarian cyst on the response to controlled ovarian hyperstimulation and the outcome of IVF-ET. One hundred one patients who underwent IVF-ET were enrolled in this study. The outcome of 31 patients, who had an ovarian cyst of >10mm detected at ultrasound examination performed on day 3, was compared with that of 70 patients who underwent a similar protocol and did not have an ovarian cyst. E2 level on the day of hCG administration, the number of follicles, the number of oocytes retrieved, the number of embryo transferred and the pregnancy rate were evaulated. The E2 level on the day of hCG adminstration and the number of mature oocytes retrieved were lower in the group with a baseline cyst. The pregnancy rate also was significantly lower in the group with a cyst (21% versus 38%). Therefore a baseline ovarian cyst on cycle day 3 was associated with a poorer outcome after IVF-ET.

• Journal of Embryo Transfer
• /
• v.9 no.2
• /
• pp.153-160
• /
• 1994

This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 $\mu$sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.

• Journal of The Korean Society of Integrative Medicine
• /
• v.11 no.2
• /
• pp.221-230
• /
• 2023

Purpose : This study was performed to evaluate the effects of virtual reality combined robot assist gait training (VRG) on improvement of balance and respiratory function in chronic stroke patients. Methods : A single-blind, randomized controlled trial (RCT) was conducted with 35 chronic stroke patients. They were randomly allocated 2 groups; VRG group (n=18) and conservative treatment group (CG; n=17). The VRG group received 30 minutes robot assisted gait training combined virtual reality training, robot assisted gait training was conducted in parallel using a virtual reality device (2 sessions of 15 minutes in a 3D-recorded walking environment and 15 minutes in a downtown walking environment). In the conservative treatment group, neurodevelopmental therapy and exercise therapy were performed according to the function of stroke patients. Each group performed 30 minutes a day 3 times a week for 8 weeks. The primary outcome balance and respiratory function were measured by a balance measurement system (BioRescue, Marseille, France), Berg balance scale, functional reach test for balance, Spirometry (Cosmed Micro Quark, Cosmed, Italy) for respiratory function Forced vital capacity (FVC), forced expiratory volume in 1 second (FEV₁), and maximum expiratory volume (PEF) were measured according to the protocol. The measurement were performed before and after the 8 weeks intervention period. Results : Both groups demonstrated significant improvement of outcome in balance and respiratory function during intervention period. VRG revealed significant differences in balance and respiratory function as compared to the CG groups (p<.05). Our results showed that VRG was more effective on balance and respiratory function in patients with chronic stroke. Conclusion : Our findings indicate that VRG can improve balance and respiratory function, highlight the benefits of VRG. This study will be able to be used as an intervention data for recovering balance and respiratory function in chronic stroke patients.

• Journal of Embryo Transfer
• /
• v.29 no.3
• /
• pp.235-240
• /
• 2014

Ethylene glycol (EG) has been successfully used as a cryoprotectant for vitrification of mammalian embryos (including human embryos) due to its low formula weight and high permeation into cells compared with other cryoprotectants, including propylene glycol (PROH). Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Female ICR mice (6~8 weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48 h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only during cryoprotectant step (1~4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows : There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3 and 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.