The present study was carried out to develop a cloning technology of mouse embryos by nuclear transplantation with electrofusion and to produce cloned offsprings by transfer of reconstituted embryos. A single nucleus from two- and eight-cell embryos was transplanted into the enucleated two-cell embryos by rnicromanipulation. The fusion of nucleus with recipient cytoplasm and the subsequent development of reconstituted embryos in vitro as well as in vivo to term were examined to determine the optimal electrofusion parameters for nuclear transplantation in mouse embryos. The successful enucleation of donor embryos was 84.9 and 83.3% in two- and eight-cell stage, respectively, and the successful injection of nucleus from two- and eight-cell donor embryos into the perivitelline space of enucleated two-cell embryos were 85.1 and 84.7%, respectively. No significant differences were found in enucleation or injection rate between the cell stages of donor embryos. When the blastomeres of intact two-cell mouse embryos were electrofused in 0.3 M mannitol medium(100 $\mu$sec., 3 pulses), the fusion rate was similarly 93.2, 92.2 and 92.0% in 1.0, 1.5 and 2.0 kV /crn, respectively, but in vitro development to blastocyst of the fused two-cell embryos was significantly(P<0.05) lower in 2.0 kV/cm (63.4%) than in 1.0 kV/cm (91.7%) or 1.5 kV/cm (82.4%). The development in vitro to eight-cell stage of the reconstituted embryos with nucleus from two-cell stage(45.5%) was significantly(P<0.05) higher than that from eight-cell stage blastomeres (16.7%). The number of blastomeres of the intact embryos at blastocyst stage was 50i0.6 and 55$\pm$2.4 in in vitro and in vivo cultured mouse embryos, respectively, but significantly(P<0.05) decreased to 35$\pm$0.7 in nuclear transplanted blastocyst embryos. The conception rate of mice following embryo transfer was 32.1% in the reconstituted two-cell embryos using two-cell donor nuclei, which was comparable to the fresh two-cell embryos(40.6%). However, the rate of development in vivo to term following embryo transfer of the reconstituted two-cell embryos using two-cell donor nuclei (23.5%) was significantly(P<0.05) lower compared with the percentage of two-cell fresh embryos(31.5%).
The Wnt/${\beta}$-catenin signaling pathway regulates diverse developmental processes and adult tissue homeostasis. Inappropriate regulation of this pathway has been associated with human diseases, such as cancers, osteoporosis, and Alzheimer's disease. Using a cell-based chemical screening with natural compounds, we discovered silybin, a plant flavonoid isolated from the Silybum marianum, which activated the Wnt/${\beta}$-catenin signaling pathway in a synergy with Wnt3a-conditioned medium (Wnt3a-CM). In the presence of Wnt3a-CM, silybin up-regulated ${\beta}$-catenin response transcription (CRT) in HEK293-FL reporter cells and 3T3-L1 preadipocytes through stabilization of intracellular ${\beta}$-catenin protein. Silybin and Wnt3a-CM synergistically reduced expression of important adipocyte marker genes including peroxisome-proliferator-activated $receptor{\gamma}$ ($PPAR{\gamma}$) and CAATT enhancer-binding protein ${\alpha}$ (C/$EBP{\alpha}$) in 3T3-L1 preadipocytes, accompanied by the activation of Wnt/${\beta}$-catenin signaling pathway. Taken together, our findings indicate that silybin is a small-molecule synergist of the Wnt/${\beta}$-catenin signaling pathway and can be used as a controllable reagent for investigating biological processes that involve the Wnt/${\beta}$-catenin signaling pathway.
Ha, Su-Min;Hwang, Shin;Park, Jin Young;Lee, Young-Joo;Kim, Ki-Hun;Song, Gi-Won;Jung, Dong-Hwan;Yu, Yun-Suk;Kim, Jinpyo;Lee, Kyoung-Jin;Tak, Eunyoung;Park, Yo-Han;Lee, Sung-Gyu
Annals of Surgical Treatment and Research
/
v.95
no.6
/
pp.303-311
/
2018
Purpose: OncoHepa test is a multigene expression profile test developed for assessment of hepatocellular carcinoma (HCC) prognosis. Multiplication of ${\alpha}$-FP, des-${\gamma}$-carboxy prothrombin (DCP) and tumor volume (TV) gives the ${\alpha}$-FP-DCP-volume (ADV) score, which is also developed for assessment of HCC prognosis. Methods: The predictive powers of OncoHepa test and ADV score were validated in 35 patients who underwent curative hepatic resection for naïve solitary HCCs ${\leq}5cm$. Results: Median tumor diameter was 3.0 cm. Tumor recurrence and patient survival rates were 28.6% and 100% at 1 year, 48.6% and 82.9% at 3 years, and 54.3% and 71.4% at 5 years, respectively. The site of first tumor recurrence was the remnant liver in 18, lung in 1, and the peritoneum in 1. All patients with HCC recurrence received locoregional treatment. OncoHepa test showed marginal prognostic significance for tumor recurrence and patient survival. ADV score at 4log also showed marginal prognostic difference with respect to tumor recurrence and patient survival. Combination of these 2 tests resulted in greater prognostic significance for both tumor recurrence (P = 0.046) and patient survival (P = 0.048). Conclusion: Both OncoHepa test and ADV score have considerably strong prognostic power, thus individual and combined findings of OncoHepa test and ADV score will be helpful to guide postresection surveillance in patients with solitary HCCs ${\leq}5cm$.
The Wetting of fusion cast $Al_2O_3$ brick and $Al_2O_3 -ZrO_2$ brick by liquid Ag was studied by the sessile drop technique in Ar atmosphere. In this experiment the specimens were photographed per 2$0^{\circ}C$with increasing temperature from 96$0^{\circ}C$ melting point of Ag. And the method of photographing was carried out by shadow technique. The cosine of the contact angle increased linearly with increasing temperature in both systems. And the relation between the cosine of the contact angle and the temperature was Cos$\theta$=1.132+$0.75{\times}10^{-3}T$ for $Al_2O_3$ brick and Cos$\theta$=-1.706+$1.125{\times}10^{-3}T$ for $Al_2O_3 -ZrO_2$ brick In both systems the contact angle decreased as the surface of substrate became smoother. The work of adhesion which was 503.5ergs/$cm^2$ for $Al_2O_3$brick and 393.6 ergs/cm2 for $Al_2O_3 -ZrO_2$ brick at 96$0^{\circ}C$ increased parabolically with increasing temperature in both system.
Foraminal decompression using a minimally invasive technique to preserve facet joint stability and function without fusion reportedly improves the radicular symptoms in approximately 80% of patients and is considered one of the good surgical treatment choices for lumbar foraminal or extraforaminal stenosis. However, proper decompression was not possible because of the inability to access the foramen at the L5-S1 level due to prominence of the iliac crest. To overcome this challenge, endoscopy-based minimally invasive spine surgery has recently gained attention. Here, we report the technical skills required in unilateral extraforaminal biportal endoscopic spinal surgery using a $30^{\circ}$ arthroscope to enable foraminal decompression at the L5-S1 level. Two 0.8-cm portals were created 2 cm lateral from the lateral border of the pedicles at the L5-S1 level. After sufficient working space was made, half of the superior articular process (SAP) in the hypertrophied facet joint was removed using a high-speed burr and a 5-mm wide osteotome, whereas the remaining inside part of the SAP was removed using a Kerrison punch and pituitary punch. The foraminal ligamentum flavum should be removed to inspect the conditions of the L5 exiting root and disc. Removing of the extruded disc could decompress the L5 root. The extraforaminal approach using a $30^{\circ}$ arthroscope is considered a minimally invasive alternative technique for decompressing foraminal stenosis at the L5-S1 level that preserves facet stability and provides symptomatic relief.
This study was designed to determine the most effective concentration of fluoride and energy density of laser irradiation for the anticarious effect. For this study surface hardness in enamel was measured before and after irradiation with pulsed Nd;YAG laser and the topical application of fluoride. Of the permanent mandibular anterior hovine teeth, healthy, carious free ones were used. Three hundred specimens were made. Specimens within 25~45 Vickers hardness numbers were assigned to 20 control and experimental stoops ; each containing 15 specimens. After forming artificial carious lesions, 10 J/$\textrm{cm}^2$, 20 J/$\textrm{cm}^2$, and 30 J/$\textrm{cm}^2$ energies were irradiated on the enamel surface of each experimental group. Also NaF, NH4F, Elmex gel(amine fluoride) and APF gel fluoride compounds were applied topically. Next, all the specimens were placed into the pH circulatory procedures for eight days. Vickers hardness numbers were measured using a microhardness tester. Surface changes of the enamel were observed using an scanning electron microscope. The comparative ana1ysis yielded the following results : 1. The reduction of surface hardness of the enamel surface was less in all groups with fluoride application than in the group without fluoride application. 2. The APF gel croup with 10 J/$\textrm{cm}^2$ irradiation showed the lowest reduction of surface hardness. 3. The reduction of surface hardness of the enamel surface in the group of laser irradiation without fluoride application not showed any significant difference according to the energy density of the laser. 4. Under the scanning electron microscope, in enamel irradiated with 10J/$\textrm{cm}^2$ showed appearance similar to acid etching surface. In enamel irradiated with 20 J/$\textrm{cm}^2$, line enamel crack was detected. In enamel irradiated with 30j/$\textrm{cm}^2$, severe enamel crack and fusion of enamel were detected. These results suggest that one could obtain the best anticariogenic effects without damage to teeth in the group of application of APF gel after laser irradiation with 10 J/$\textrm{cm}^2$.
In this study, we measured Near Point of Convergence(N.P.C) tests, Phoria tests using Von Grafe method and relative convergence tests on 138 men and 162 women, so a total of 300 subjects aged between 8~45 to examine the improvement of the fusion vergence through visual training and obtained as follows. 1. According to the results, the near point of convergence of 57 (19%) subjects were shorter than 7cm, and 243 (81%) were 7cm or longer, having a problem in convergence. After visual training, the number of subjects have the value shorter than 7 cm increased from 57 to 111 (37%), and the number of those have the value 7cm or longer decreased significantly form 243 to 189 (63%). 2. The results of the measure of lateral Phoria at far distance by Von Grafe method showed orthophoria 18 (6%), exophoria 198 (66%), esophoria 84 (28%). After phoria test, we examined the N.R.C and P.R.C test. The results showed that the hope finger was improved after V.T using B.l, B.O card. 3. The results of the measure of lateral Phoria at near distance by Von Grafe method showed orthophoria 6 (2%), exophoria 222 (74%), esophoria 72 (24%). After phoria test, we examine the N.R.C and P.R.C test. The results showed that the hope finger was improved after V.T using B.I, B.O card.
The Journal of Korean Society for Radiation Therapy
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v.22
no.2
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pp.75-83
/
2010
Purpose: It aims to evaluate the location change and tendency of hepatic and intrahepatic tumors according to gastric volume and change of location. Materials and Methods: It studied 9 patients with hepatic tumors who visited Gangnam Severance Hospital from March 2009 to April 2010 and who underwent CT or PET (Positron Emission Tomography) within 2 weeks before CT-simulation. The patients fasted for 6 hours before CT-simulation and drank 240~250 cc of water just before CT or PET for image fusion. Those two types of images were fused to RTP (Radiation Treatment Planning, Pinnacle 8.0h) focusing on bone structure of individual patients. Results: They drank 240~260 cc of water but their stomach volume after drinking water varied from 259.3 cc to 495.4 cc. Even though individual differences existed in the change of stomach volume before and after drinking water, the volume was increased by 130 cc (174%) on average. The change in absolute distance between the centers of tumors ranged from 0.52 cm to 3.04 cm (1.52 cm on average); from 0.1 cm to 1.35 cm (0.44 cm on average) in cranial-caudal direction; from 0.05 cm to 2.75 cm (1.22 cm on average) in left-right direction; and from 0.05 cm to 1.85 cm (0.33 cm on average) in ventral-dorsal direction. Conclusion: It is hard to predict the movement of tumors by observing stomach movement, due to great individual differences; however, it was observed that the location of hepatic tumors was right-sided as the stomach was filled with water. Thus, it is recommended to maintain the fastened state to secure the accuracy of hepatic tumor treatment. If it cannot maintain the fastened state, it is recommended to measure stomach volumes and movement in the patient to consider the movement of hepatic tumors before radiation treatment.
Objective : Although unilateral transforaminal lumbar interbody fusion (TLIF) is widely used because of its benefits, it does have some technical limitations. Removal of disk material and endplate cartilage is difficult, but essential, for proper fusion in unilateral surgery, leading to debate regarding the surgery's limitations in removing the disk material on the contralateral side. Therefore, authors have conducted a randomized, comparative cadaver study in order to evaluate the efficiency of the surgery when using conventional instruments in the preparation of the disk space and when using the recently developed high-pressure water jet system, SpineJet$^{TM}$ XL. Methods : Two spine surgeons performed diskectomies and disk preparations for TLIF in 20 lumbar disks. All cadaver/surgeon/level allocations for preparation using the SpineJet$^{TM}$ XL (HydroCision Inc., Boston, MA, USA) or conventional tools were randomized. All assessments were performed by an independent spine surgeon who was unaware of the randomizations. The authors measured the areas (cm2) and calculated the proportion (%) of the disk surfaces. The duration of the disk preparation and number of instrument insertions and withdrawals required to complete the disk preparation were recorded for all procedures. Results : The proportion of the area of removed disk tissue versus that of potentially removable disk tissue, the proportion of the area of removed endplate cartilage, and the area of removed disk tissue in the contralateral posterior portion showed 74.5 ${\pm}$ 17.2%, 18.5 ${\pm}$ 12.03%, and 67.55 ${\pm}$ 16.10%, respectively, when the SpineJet$^{TM}$ XL was used, and 52.6 ${\pm}$ 16.9%, 22.8 ${\pm}$ 17.84%, and 51.64 ${\pm}$ 19.63%, respectively, when conventional instrumentations were used. The results also showed that when the SpineJet$^{TM}$ XL was used, the proportion of the area of removed disk tissue versus that of potentially removable disk tissue and the area of removed disk tissue in the contralateral posterior portion were statistically significantly high (p < 0.001, p < 0.05, respectively). Also, compared to conventional instrumentations, the duration required to complete disk space preparation was shorter, and the frequency of instrument use and the numbers of insertions/withdrawals were lower when the SpineJet$^{TM}$ XL was used. Conclusion : The present study demonstrates that hydrosurgery using the SpineJet$^{TM}$ XL unit allows for the preparation of a greater portion of disk space and that it is less traumatic and allows for more precise endplate preparation without damage to the bony endplate. Furthermore, the SpineJet$^{TM}$ XL appears to provide tangible benefits in terms of disk space preparation for graft placement, particularly when using the unilateral TLIF approach.
This study investigated whether the addition of porcine sperm cytosolic factor (SCF) at fusion/activation affects in vitro development of porcine parthenogenetic(PA) and nuclear transfer (NT) embryos. To determine the optimum concentration of SCF, control group of oocytes was activated with 0.3M mannitol (1.0 mM $CaCl_2{\cdot}2H_2O$), other three groups of oocytes were parthenogentically activated with the fusion medium (0.1mM $CaCl_2{\cdot}2H_2O$) supplemented with 100, 200 or 300 ${\mu}$g/ml SCF, respectively. Matured oocytes were activated with two electric pulses (DC) of 1.2 kv/cm for 30 ${\mu}$sec. The activated embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. Oocytes activated in the presence of SCF showed a significantly higher blastocyst rate than control (p<0.05). Apoptosis rate was significantly lower in 100 ${\mu}$g/ml SCF group than other groups (p<0.05). Cdc2 kinase activity in control and SCF treatment group of oocytes was determined using MESACUP cdc2 kinase assay kit at 1, 5, 10, 15, 30, 45 and 60 min after activation. Cdc2 kinase activity was significantly decreased (p<0.05) in SCF group than MII oocytes or control within 5 min. For NT embryo production, reconstructed oocytes were fused in the fusion medium supplemented with 0.1 mM $CaCl_2{\cdot}2H_2O$ (T1), 1.0 mM $CaCl_2{\cdot}2H_2O$ (T2) and 0.1 mM $CaCl_2{\cdot}2H_2O$ with 100 ${\mu}$g/ml SCF (T3). Fused embryos were cultured in PZM-3 under 5% $CO_2$ in air at $38.5^{\circ}C$ for 6 days. Developmental rate to blastocyst stage was significantly higher in T3 than other groups (23.0% vs. 13.5 to 15.2%) (p<0.05). Apoptosis rate was significantly lower in T3 than T1 or T2 (p<0.05). The relative abundance of Bax-${\alpha}$/Bcl-xl was significantly lower in in vivo or SCF group than that of control (p<0.05). Moreover, the expression of p53 and caspase3 mRNA was significantly lower in in vivo or SCF group than that of control (p<0.05). These results indicate that the addition of SCF at fusion/activation might improve in vitro development of porcine NT embryos through regulating cdc2 kinase level and expression of apoptosis related genes.
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