• Title/Summary/Keyword: donor cell

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Lateral Oropharyngeal Wall Coverage with Buccinator Myomucosal and Buccal Fat Pad Flaps

  • Jung, Bok Ki;Song, Seung Yong;Kim, Se-Heon;Kim, Young Seok;Lee, Won Jai;Hong, Jong Won;Roh, Tai Suk;Lew, Dae Hyun
    • Archives of Plastic Surgery
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    • v.42 no.4
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    • pp.453-460
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    • 2015
  • Background Reconstruction of oropharyngeal defects after resection of oropharyngeal cancer is a significant challenge. The purpose of this study is to introduce reconstruction using a combination of a buccinator myomucosal flap and a buccal fat pad flap after cancer excision and to discuss the associated anatomy, surgical procedure, and clinical applications. Methods In our study, a combination of a buccinator myomucosal flap with a buccal fat pad flap was utilized for reconstruction after resection of oropharyngeal cancer, performed between 2013 and 2015. After oropharyngectomy, the defect with exposed vital structures was noted. A buccinator myomucosal flap was designed and elevated after an assessment of the flap pedicle. Without requiring an additional procedure, a buccal fat pad flap was easily harvested in the same field and gently pulled to obtain sufficient volume. The flaps were rotated and covered the defect. In addition, using cadaver dissections, we investigated the feasibility of transposing the flaps into the lateral oropharyngeal defect. Results The reconstruction was performed in patients with squamous cell carcinoma. The largest tumor size was $5cm{\times}2cm(length{\times}width)$. All donor sites were closed primarily. The flaps were completely epithelialized after four weeks, and the patients were followed up for at least six months. There were no flap failures or postoperative wound complications. All patients were without dietary restrictions, and no patient had problems related to mouth opening, swallowing, or speech. Conclusions A buccinator myomucosal flap with a buccal fat pad flap is a reliable and valuable option in the reconstruction of oropharyngeal defects after cancer resection for maintaining functionality.

Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

  • Yuan, Yu-Guo;Song, Shao-Zheng;Zhu, Meng-Ming;He, Zheng-Yi;Lu, Rui;Zhang, Ting;Mi, Fei;Wang, Jin-Yu;Cheng, Yong
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1175-1182
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    • 2017
  • Objective: To create genetically modified goat as a biopharming source of recombinant human lacotoferrin (hLF) with transcription activator-like effector nucleases. Methods: TALENs and targeting vector were transferred into cultured fibroblasts to insert hLF cDNA in the goat beta-lactoglobulin (BLG) locus with homology-directed repair. The gene targeted efficiency was checked using sequencing and TE7I assay. The bi-allelic gene targeted colonies were isolated and confirmed with polymerase chain reaction, and used as donor cells for somatic cell nuclear transfer (SCNT). Results: The targeted efficiency for BLG gene was approximately 10%. Among 12 Bi-allelic gene targeted colonies, five were used in first round SCNT and 4 recipients (23%) were confirmed pregnant at 30 d. In second round SCNT, 7 (53%), 4 (31%), and 3 (23%) recipients were confirmed to be pregnant by ultrasound on 30 d, 60 d, and 90 d. Conclusion: This finding signifies the combined use of TALENs and SCNT can generate biallelic knock-in fibroblasts that can be cloned in a fetus. Therefore, it might lay the foundation for transgenic hLF goat generation and possible use of their mammary gland as a bioreactor for large-scale production of recombinant hLF.

The Inhibitory Effects of Hydrogen Sulfide on Pacemaker Activity of Interstitial Cells of Cajal from Mouse Small Intestine

  • Parajuli, Shankar Prasad;Choi, Seok;Lee, Jun;Kim, Young-Dae;Park, Chan-Guk;Kim, Man-Yoo;Kim, Hyun-Il;Yeum, Cheol-Ho;Jun, Jae-Yeoul
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.2
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    • pp.83-89
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    • 2010
  • In this study, we studied whether hydrogen sulfide ($H_2S$) has an effect on the pacemaker activity of interstitial cells of Cajal (ICC), in the small intestine of mice. The actions of $H_2S$ on pacemaker activity were investigated using whole-cell patch-clamp technique, intracellular $Ca^{2+}$ analysis at $30^{\circ}C$ and RT-PCR in cultured mouse intestinal ICC. Exogenously applied sodium hydrogen sulfide (NaHS), a donor of hydrogen sulfide, caused a slight tonic inward current on pacemaker activity in ICC at low concentrations (50 and $100{\mu}m$), but at high concentration ($500{\mu}m$ and 1 mM) it seemed to cause light tonic inward currents and then inhibited pacemaker amplitude and pacemaker frequency, and also an increase in the resting currents in the outward direction. Glibenclamide or other potassium channel blockers (TEA, $BaCl_2$, apamin or 4-aminopydirine) did not have an effect on NaHS-induced action in ICC. The exogenous application of carbonilcyanide p-triflouromethoxyphenylhydrazone (FCCP) and thapsigargin also inhibited the pacemaker activity of ICC as NaHS. Also, we found NaHS inhibited the spontaneous intracellular $Ca^{2+}$ ($[Ca^{2+}]_i$) oscillations in cultured ICC. In doing an RT-PCR experiment, we found that ICC enriched population lacked mRNA for both CSE and CBS, but was prominently detected in unsorted muscle. In conclusion, $H_2S$ inhibited the pacemaker activity of ICC by modulating intracellular $Ca^{2+}$. These results can serve as evidence of the physiological action of $H_2S$ as acting on the ICC in gastrointestinal (GI) motility.

Development of aortic endothelial cells to express CD37 and CD73 isolated from alpha 1,3-galactosyltransferase knock-out and MCP expressing pig (alpha 1,3-galactosyltransferase 기능 제거 및 MCP 발현 형질전환 돼지의 대동맥 혈관내피세포에 CD37/CD73 발현 세포주 개발)

  • No, Jin-Gu;Byun, Sung-June;Yang, Hyeon;Ock, Sun A;Woo, Jae-Seok;Lee, Hwi-Cheul;Hwang, In-sul;Kim, Ji-Youn;Park, Sang Hyoun;Lee, Joo Young;Oh, Keon Bong
    • Journal of Embryo Transfer
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    • v.33 no.3
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    • pp.129-137
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    • 2018
  • Acute vascular rejection has been known as a main barrier occurring in a xenograted tissue of alpha 1,3-galactosyltransferase knock-out (GalT KO) pig into a non-human primate (NHP). Adenosine which is a final metabolite following sequential hydrolysis of nucleotide by ecto-nucleotidases such as CD39 and CD73, act as a regulator of coagulation, and inflammation. Thus xenotransplantation of CD39 and CD73 expressing pig under the GalT KO background could lead to enhanced survival of recipient NHP. We constructed a human CD39 and CD73 expression cassette designed for endothelial cell-specific expression using porcine Icam2 promoter (pIcam2-hCD39/hCD73). We performed isolation of endothelial cells (pAEC) from aorta of 4 week-old GalT KO and membrane cofactor protein expressing pig ($GalT^{-MCP/-MCP}$). We were able to verify that isolated cells were endothelial-like cells using immunofluorescence staining analysis with von Willebrand factor antibody, which is well known as an endothelial maker, and tubal formation assay. To find optimal condition for efficient transfection into pAEC, we performed transfection with GFP expression vector using four programs of nucleofection, M-003, U-023, W-023 and Y-022. We were able find that the program W-023 was optimal for pAEC with regard to viability and transfection efficiency by flow cytometry and fluorescent microscopy analyses. Finally, we were able to obtain $GalT^{-MCP/-MCP}/CD39/CD73$ pAEC expressing CD39 and CD73 at levels of 33.3% and 26.8%, respectively. We suggested that pACE isolated from $GalT^{-MCP/-MCP}$ pig might be provided as a basic resource to understand biochemical and molecular mechanisms of the rejections and as an alternative donor cells to generate $GalT^{-MCP/-MCP}/CD39/CD73$ pig expressing CD39 and CD73 at endothelial cells.

Intraspecific Androgenesis in Mud Loach (Misgurnus mizolepis): II. Diploid Restoration and Viability Assessment (미꾸라지(Misgurnus mizolepis)의 웅성발생성 처녀생식: II. 웅성발생성 이배체 유도 및 생존능력 평가)

  • Nam, Yoon-Kwon;Bang, In-Chul;Noh, Choong-Hwan;Cho, Young-Sun;Kim, Dong-Soo
    • Journal of Aquaculture
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    • v.19 no.4
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    • pp.281-287
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    • 2006
  • Intraspecific diploid androgenesis was achieved in mud loach (Misgurnus mizolepis) by the inhibition of the first mitotic division using combined thermal treatment. A combined thermal treatment (heat shock at $40.5\;^{\circ}C$ for 120 sec followed by cold treatment at $1\;^{\circ}C$ for 45 min) applied to the 1st metaphase of cell division (28 min post insemination at $25\;^{\circ}C$) successfully recovered viable androgenetic diploidy. Mean hatching success of the androgenetic diploid group was 29.6%, and the average yield out of total eggs taken was about 7% assessed at 1 week of age. However, relatively large variations in the yield of diploid androgenesis were observed among different egg batches used as cytoplasmic donors. Successful diploidization was confirmed by flow cytometric analysis, and parthenogenic reproduction in a paternal exclusive manner was verified with transgene dosage. Significant mortality was found in most androgenetic groups especially from hatch to 1 month of age, although such mortality was stabilized later.

Treatment of Benign Bone Tumor with Xenograft (이종골 이식을 이용한 양성 골 종양 치료)

  • Kim, Han-Soo
    • The Journal of the Korean bone and joint tumor society
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    • v.1 no.2
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    • pp.189-193
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    • 1995
  • The authors reviewed 49 cases(48 patients) of benign bone tumor who had surgical treatment with xenograft at department of orthopedic surgery, Seoul National University Hospital from May, 1980 to May, 1994. Materials consist of 21 males and 27 females. The mean age at operation was 20.1 years(range : 4 -55 years) and the mean follow up period was 25.4 months(range : 7 - 85 months). We did xenograft only in 34 cases and xenograft mixed with autograft in 15 cases(14 cases, from ilium, 1 case from femur). The used materials for xenograft were $Lubboc^{(R)}$ in 29 cases, $Surgibone^{(R)}$ in 17 cases and $Pyrost^{(R)}$ in 3 cases. The average durations when bony union was achieved in radiograph were 13.8 weeks in whole cases, 12.5 weeks(range : 8 - 24 weeks) in $Lubboc^{(R)}$ graft cases and 15.7 weeks(range : 6 - 24 weeks) in $Surgibone^{(R)}$ graft cases. The tumor recurred in 4 cases, 1 case was recurred giant cell tumor at distal femur treated with mixed auto and $Lubboc^{(R)}$ graft and 2 cases were large cystic lesions at the proximal humerus diagnosed as simple bone cyst and at distal tibia diagnosed as fibrous dysplasia treated with $Surgibone^{(R)}$ graft and 1 case was aneurysmal bone cyst of the proximal tibia treated with $Lubboc^{(R)}$ graft. Wound infection occurred in 1 case. More transfusion was done in the cases that the lesion was larger than 5cm, the lesions were in the ilium or femur and the cases that were treated with mixed with autograft. This study implies that benign bone tumor is successfully treqated with curettage and xenograft or xenograft mixed with autograft. And also this method will reduce morbidity of donor site, intraoperative bleeding and post-transfusion complications.

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Experimental Study of Isolated Canine Heart Preservation for 24 Hours at 4$^{\circ}C$ - A Portable Continuous Hypothermic Perfusion System - (적출심장의 장시간 [24시간] 보존에 관한 실험적 연구 -4$^{\circ}C$ 관류 보존법-)

  • 이종국
    • Journal of Chest Surgery
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    • v.21 no.3
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    • pp.425-446
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    • 1988
  • After 24 hours of preservation under 15 mmHg perfusion pressure the recovery rates of isolated canine hearts were determined. Preservation was performed in a cold room maintained at 4*C with 4 different types of perfusates bubbled with a mixture of 95% 0y and 5% CO~ using a modified perfusion unit designed in our institute. The perfusates used were as follows; Group 1: Krebs-Henseleit solution, Group 2: Krebs solution added by albumin and PGE1. Group 3: Modified Wicomb*s solution, Group 4: Modified Collin*s solution. The extent of myocardial recovery was evaluated using a modified isolated carmine perfusion model by measuring heart rate, systolic arterial pressure, left atrial pressure[LAP] and cardiac output. In addition to the above hemodynamic parameters, biochemical and enzymatic assays from perfusates and electron microscopic changes of the myocardium were also studied. The results were as follows; 1] The heart recovery rates were 41.6%, 53.4% and 108.9% in groups 1, 2 and 3, respectively, and group 3 elicited the best result[p< 0.001]. The heart beat was never recovered in group 4. 2] Recovered systolic arterial pressures[mmHg] were 63.3% in group 1, 94.9% in group 2 and 94.3% in group 3. 3] LAPs[mmHg] were 20 in group 1, 13.5 in group 2 and 11.2 in group 3, which suggested that the best myocardial preservation was elicited in group 3[p< 0.05]. 4] Cardiac output, the sum of aortic stroke volume and coronary leakage, were 69.1% in group 2, and 90.7% in group 3, but these were not statistically significant[p=0.24]. No aortic stroke output was measured in group 1 and 4. 5] The degree of myocardial edema increase was 17.5` in group 1, 24.6% in group 2, 20.9% in group 3 and 55.3% in group 4. But there were no statistical differences in each group[p= 0.08]. 6] CPK-MB[U/L] levels were increased 750% and 332%[p< 0.05], glucose levels[mg/dl] 60.5% and 78.2% and SGOT[U/L] levels 523% and 333%, in groups 2 and 3, respectively. Biochemical and enzymatic assays could not be performed in group 1 and group 4, because of poor recovery of heart beat. 7] Electron microscopic findings in the myocardium of most groups revealed slight to moderate muscle cell and mitochondrial edema. But all these findings were within the limits of reversible change. From these above results, it is suggested that modified Wicomb*s solution seems to be the most useful physiologic salt solution for preservation of the heart. We propose that after further study and improvement, our portable continuous hypothermic perfusion system will contribute to the development of a better preservation method for donor hearts for human heart transplantation.

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Growth and optical characterization of $CuInSe_2$ single crystal thin film for solar cell application (태양전지용 $CuInSe_2$단결정 박막 성장과 광학적 특성)

  • 백승남;홍광준
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.12 no.4
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    • pp.202-209
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    • 2002
  • The stochiometric mix of evaporating materials for the $CuInSe_2$single crystal thin films was prepared from horizontal furnace. To obtain the single crystal thin films, $CuInSe_2$compound crystal was deposited on thoroughly etched semi-insulating GaAs(100) substrate by the Hot Wall Epitaxy (HWE) system. The source and substrate temperature were $620^{\circ}C$ and $410^{\circ}C$, respectively. The crystalline structure of single crystal thin films was investigated by the photoluminescence and double crystal X-ray diffraction (DCXD). The carrier density and mobility of $CuInSe_2$single crystal thin films measured from Hall effect by van der Pauw method. From the photocurrent spectrum by illumination of perpendicular light on the c-axis of the $CuInSe_2$single crystal thin film, we have found that the values of spin orbit splitting $\Delta$So and the crystal field splitting $\Delta$Cr. From the photoluminescence measurement on $CuInSe_2$single crystal thin film, we observed free exciton ($E_x$) existing only high quality crystal and neutral bound exciton ($A^{\circ}$, X) having very strong peak intensity. Then, the full-width-at-half-maximum (FWHM) and binding energy of neutral donor bound exciton were 7 meV and 5.9 meV, respectivity. By haynes rule, an activation energy of impurity was 59 meV.

Synthesis and Characterization of Quinoxaline-Based Thiophene Copolymers as Photoactive Layers in Organic Photovoltaic Cells

  • Choi, Yoon-Suk;Lee, Woo-Hyung;Kim, Jae-Ryoung;Lee, Sang-Kyu;Shin, Won-Suk;Moon, Sang-Jin;Park, Jong-Wook;Kang, In-Nam
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.417-423
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    • 2011
  • A series of new quinoxaline-based thiophene copolymers (PQx2T, PQx4T, and PQx6T) was synthesized via Yamamoto and Stille coupling reactions. The $M_ws$ of PQx2T, PQx4T, and PQx6T were found to be 20,000, 12,000, and 29,000, with polydispersity indices of 2.0, 1.2, and 1.1, respectively. The UV-visible absorption spectra of the polymers showed two distinct absorption peaks in the ranges 350 - 460 nm and 560 - 600 nm, which arose from the ${\pi}-{\pi}^*$ transition of oligothiophene units and intramolecular charge transfer (ICT) between a quinoxaline acceptor and thiophene donor. The HOMO levels of the polymer ranged from -5.37 to -5.17 eV and the LUMO levels ranged from -3.67 to -3.45 eV. The electrochemical bandgaps of PQx2T, PQx4T, and PQx6T were 1.70, 1.71, and 1.72 eV, respectively, thus yielding low bandgap behavior. PQx2T, PQx4T, and PQx6T had open circuit voltages of 0.58, 0.42, and 0.47 V, and short circuit current densities of 2.9, 5.29 and 9.05 mA/$cm^2$, respectively, when $PC_{71}BM$ was used as an acceptor. For the solar cells with PQx2T-PQx6T:$PC_{71}BM$ (1:3) blends, an increase in performance was observed in going from PQx2T to PQx6T. The power conversion efficiencies of PQx2T, PQx4T, and PQx6T devices were found to be 0.69%, 0.73%, and 1.80% under AM 1.5 G (100 mW/$cm^2$) illumination.

Apoptosis and Peripheral Benzodiazepin Receptor (PBR) Expression in Human Granulosa-Luteal Cells by GnRH-agonist (GnRH-agonist에 의한 인간 과립-황체화 세포의 세포사멸과 PBR 단백질의 발현)

  • Kim, Sei-Kwang;Youm, Yun-Hee;Yoon, Jeong-Mi;Bai, Sang-Wook;Yang, Hyun-Won;Cho, Dong-Jae;Yoon, Yong-Dal;Song, Chan-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.2
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    • pp.83-94
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    • 2004
  • Objective: To investigate whether GnRH-agonist (GnRH-Ag) using in IVF-ET affects apoptosis of human granulosa-luteal cells and expression of peripheral benzodiazepine receptor (PBR) protein involved in the apoptosis of the cells. Methods: Granulosa-luteal cells obtained during oocyte retrieval were cultured and treated with $10^{-5}M$ GnRH-Ag. Apoptosis of the cells by the treatment was confirmed using DNA fragmentation analysis 24 h after culture. The presence of PBR protein within the cells was examined by immunofluorescence staining and the expression of the protein was analyzed by Western blotting. In addition, it was measured for progesterone and nitric oxide (NO) produced by granulosa-luteal cells after GnRH-Ag treatment. To evaluate the relationship between NO production and PBR expression, sodium nitroprusside (SNP) as a NO donor was added in media and investigated the expression of PBR protein by Western blotting. Results: Apoptosis increased in the granulosa-luteal cells 24 h after GnRH-Ag treatment, whereas the expression of PBR protein significantly decreased. Furthermore, the production of progesterone and nitric oxide (NO) by the cells significantly fell from 12 h after the treatment. In the results of Western blotting after SNP treatment, the expression of PBR protein increased in the treatment with SNP alone to the granulosa-luteal cells, but was suppressed in the treatment with GnRH-Ag and SNP. Additionally, the staining result of PBR protein in the cells showed the even distribution of it through the cell. Conclusion: These results demonstrate that GnRH-Ag treatment induces apoptosis, decreasing expression of PBR protein and NO production in human granulosa-luteal cells. The present study suggests that one of the apoptosis mechanism of human granulosa-luteal cells by GnRH-Ag might be a signal transduction pathway via NO and PBR.