This study was carried out to examine the effects of epidermal growth factor (EGF) and the number of cumulus-oocyte complexes (COCs) on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). COCs were collected from antral follicles of porcine ovaries collected from abattoir, and were maturated in modified NCSU-23 (mNCSU-23) with 10% pFF, 0.6 mM cysteine, 50 ${\mu}mM{\beta}-mercaptoethanol$, 1 mM dbcAMP, 10 IU/mL PMSG and 10 IU/mL hCG, which was supplemented with or without 10 ng/mL EGF and into which 50 or 15 COCs per droplet was put. Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium (mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU- 23. The results are as follows. 1.In the result of IVM, 10 ng/mL EGF supplement duplicated the percentage of C4 group of COCs(41% vs 81%). But the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, and there was not a significant interaction between the two factors, either. 2. In the result of IVF, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet, or was not a significant interaction between the two factors, in the rate of sperm penetration, in the percentage of oocytes with male pronucleus (MPN), and in the rate of polyspermy. 3. In the result of IVD, there was not significantly different between control and EGF supplemented, or between the number of COCs per culture droplet in the percentage of cleaved oocytes. There was not significantly different between the number of COCs per culture droplet, but between control and EGF supplemented (p<0.01) in the percentage of blastocysts, the number of inner cell mass (ICM), trophectoderm (TC) and total cells. There was no significant interaction between the two factors anywhere. These results suggested that 10 ng/mL EGF supplement into mNCSU-23 for IVM was effective in the production of more as well as better blastocysts during IVD through increasing the number of cells in those.
Background: Paravalvular leakage or false aneurysm developed after isolated aortic valve replacement(AVR) for aortic regurgitation(AR) associated with Behcet's disease is one of the most serious complications, and requires subsequent reoperations. We describe the surgical result of homograft aortic root replacement(ARR) for AR associated with Behcet's disease. Material and Method: From January 1992 to December 2001, 6 patients with AR associated with Behcet's disease underwent 7 ARR with homograft and 1 Ross operation. Five patients were male and one was female. The grafts used for ARR were 5 aortic and 2 pulmonic homografts. Ages at operation ranged from 27 to 51 years(mean, 37$\pm$9 years). Two patients underwent ARR with aortic homograft at the first operation. In the remaining 4 patients, ARR using a homograft was performed for paravalvular leakage that developed after AVR, and the mean interval from AVR to ARR was 21 $\pm$29 months(range, 5 to 73.3 moths, median, 7.6 months). Result: There was no early death. All patients were followed up for an average of 18.9$\pm$24.0 months(range, 1.9 to 68.9 months, median, 8.4 months). Two of 4patients who had undergone ARR after AVR required subsequent reoperations for false aneurysm of the ascending aorta and failure of pulmonary homograft. One patient underwent re-replacement of the aortic root, ascending aorta and partial aortic arch with an aortic homograft, the other underwent Ross operation. Conclusion: This study suggests that aortic root replacement using a homograft in aortic regurgitation with Behcet's disease may provide good clinical results and decrease the incidence of paravalvular leakage or false aneurysm after aortic valve replacement. However, the adequate perioperative management and complete removal of the inflarrunatory tissue at operation were also important for the good long-term results.
Kim Jae Hyun;Oh Sam Sae;Lee Chang-Ha;Baek Man Jong;Kim Chong Whan;Na Chan-Young
Journal of Chest Surgery
/
v.38
no.3
s.248
/
pp.197-203
/
2005
Homograft aortic valve replacement (AVR) has many advantages such as excellent hemodynamic performance, faster left ventricular hypertrophy regression, resistance to infection and excellent freedom of thromboembolism. To find out the results of homograft AVR, we reviewed our surgical experiences. Material and Method: Eighteen patients (male female=16 : 2, mean age=39.3$\pm$16.2 years, range: 14$\~$68 years) who underwent homo-graft aortic valve replacement between May 1995 and May 2004 were reviewed. The number of homografts was 20 (17 aortic and 3 pulmonic homografts) including two re-operations. Ten patients had a history of previous aortic valve surgery. Indications for the use of a homograft were native valve endocarditis (n=7), prosthetic valve endocarditis (n=5), or Behcet's disease (n=8). The homograft had been implanted predominantly as a full root except in one patient in the subcoronary position. Result: Mean follow-up was 41.3 $\pm$ 26.2 months. There was one operative mortality. Postoperative complications included postoperative bleeding in 3 patients, and wound infection in 1. There was no late death. Three patients underwent redo-AVR. The etiology of the three reoperated patients was Behcet's disease (p=0.025). Freedom from reoperation was $87.5\pm8.3\%$, $78.8\pm11.2\%$ at 1, 5 years respectively, In patients with infective endocarditis, there was no recurrence of endocarditis. There was no thromboembolic complication. Conclusion: Although longer term follow-up with larger numbers of patients is necessary, the operative and mid-term results for homograft AVR was good when we took into account the operative risks of Behcet's disease or infective endocarditis. Behest's disease was a risk factor for reoperation after the homograft AVR. We think homograft AVR is the procedure of choice, particularly in patients with infective endocarditis.
Kim, Hyoung-Soo;Park, Seung-Rim;Kang, Joon-Soon;Lee, Woo-Hyeong;Kim, Ki-Wook
Journal of the Korean Arthroscopy Society
/
v.5
no.1
/
pp.1-6
/
2001
Purpose : The purpose of this study was to compare the postoperative success and stability of arthroscopically assisted anterior cruciate ligament(ACL) reconstructions using central one third bone-patellar-tendon bone(BPB) autograft versus quadrupled semitendinos-us(ST) autograft as the medium term review. Materials and Methods : Eighty patients(40 BPBs,40 STs) with isolated ACL injury were available for a mean follow up of 49.4 months(BPB) and 48.8 months(ST). There was no significant difference between the two groups with respect to age and sex. We compare the final results between two groups with respect to subjective Lysholm score, objective laxity including anterior drawer test, Lachman test, pivot shift test, KT-2000 measurements and International Knee Documentation Committee(IKDC) evaluation system. Results : Postoperatively, positive anterior drawer test was found in $22.5\%\;and\;27.5\%$, positive Lachman test was found in $30.0\%$ and $25.0\%$ and positive pivot shift test was found in $15\%\;and\;20\%$ of the ST and BPB group, respectively(p>0.05). Mean side to side difference of KT-2000 at 20 lbs was 2.2 mm for the ST group and 2.1 mm for the BPB group. There was no significant difference between the two groups about Lysholm score(>0.05). Anterior knee pain was knee common in the BPB group. Eighty-three percent of the patients were nearly normal according to the IKDC grade in both groups. Conclusion : We consider that autogenous semitendinosus tendon is a good alternative subsititute in ACL reconstruction together with the bone-patellar tendon-bone autograft.
Bank, Won-Jong;Rhee, Seung-Koo;Kang, Yong-Koo;Kwon, Oh-Soo;Chung, Yang-Guk
The Journal of the Korean bone and joint tumor society
/
v.9
no.1
/
pp.124-129
/
2003
Purpose: To analyze the clinical outcome and radiological features after surgical treatment of stage III giant cell tumor around the knee. Materials and Methods: 21 patients with stage III giant cell tumor around the knee joint, who were operated at our institutes between March 1991 and February 2000, were selected for this study. The average follow-up was 5.7 years (range, 1~9 years). After thorough curettage using high speed burr, cryosurgery and cementing with polymethymethacrylate (PMMA) were performed in 11 patients. 7 patients were treated with PMMA cementing (4 patients) or bone grafting (3 patients) after curettage without cryosurgery. Reconstruction with prosthesis composite allograft and knee fusion with Huckstep nail were performed in 3 patients with huge defect and joint perforation. Results: Local recurrence developed in 1 out of 11 patients who was treated with curettage and cementing with cryosurgery (9.1%) and 3 out of 7 patients who underwent curettage and cementing without cryosurgery (28.6%). Joint space narrowing more than 3mm was noted in 1 patient (9.1%), who treated with cryosurgery and anther patient (14.5%) who treated without cryosurgery. There was no local recurrence in case of wide resection and reconstruction. Conclusion: Thorough curettage and PMMA cementing with cryosurgery as an adjuvant is thought to be effective modalities in the treatment of stage 3 giant cell tumors around the knee. Wide resection and reconstruction can be reserved mainly for the cases of stage 3 giant cell tumor with significant cortical destruction and marked joint destruction, and the cases of local recurrence with poor bone stock.
Vitrification of oocytes has been applied recently fur pigs, but remains elusive. The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine oocytes. When immature follicular oocytes frozen-thawed were cultured for in vitro maturation, maturation rates to metaphase-II stage were higher in oocytes with (25%) than without (15%) cumulus cells. After In vitro fertilization of oocytes frozen-thawed, the maturation rates were also significantly (P<0.05) higher in oocytes with (41%) that than without (17%) cumulus cells. However, the penetration rates were higher in oocytes without (19%) that than with (9%) cumulus. In another experiment, porcine oocytes matured in vitro were frozen and thawed for in vitro fertilization. The penetration rates were higher than in oocytes without (35%) that than with (26%) cumulus cells. However, the proportions of oocytes dead after in vitro fertilization were significantly (P<0.05) higher in oocytes with that than without cumulus cells. On the other hand, the rates of penetration and dead oocytes at 6 h after in vitro fertilization were not significant differences between oocytes with and without cumulus cells. However, the proportions of dead oocytes with (18%) and without (16%) cumulus cells were higher than in oocytes of control group (0%). These finding indicated the possible broader application for OPS, as they demonstrated that the maturation and fertilization in vitro by frozen-thawed oocytes may be accompained by cumulus cells and culture periods according to the requirements of the survival ability after freezing of mature and immature oocytes in pigs.
Kim Y.S.;Song S.H.;Cho S.K.;Kwack D.O.;Kim C.W.;Park C.S.;Chung K.H.
Journal of Embryo Transfer
/
v.21
no.2
/
pp.101-107
/
2006
The objective of this study was to investigative the effects of retinol supplement to IVM and/or IVC medium on maturation, fertilization and development of pig oocytes. North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (pFF) was used as base medium. Each 1 uM, 5 uM and 10 uM concentration of retinol was supplemented to IVM and /or IVC medium. When the retinol was supplemented to maturation medium, the maturation rates were not different (p>0.05) among treatment groups ($66.7{\pm}6.0{\sim}69.2{\pm}5.3%$), but the developmental rate to blastocyst stage was higher (p<0.05) in $5{\mu}M$ group ($20.4{\pm}2.6%$) than in 0 uM ($13.6{\pm}2.1%$) and 10 uM groups ($9.7{\pm}1.7%$). Moreover, total cell number was significantly greater (p<0.05) in the 5 uM group ($37.0{\pm}1.6$) than in the other groups ($29.8{\pm}1.0{\sim}33.2{\pm}1.0$). Retinol supplement to maturation medium did not significantly affect the rates of fertilization and polyspermy (p>0.05). When the retinol was supplemented to culture medium or both maturation and culture medium, the rates of cleavage, and develop to morula and blastocyst stage were not affected, while those of 10 uM group were significantly decreased (p<0.05). These results indicate that 5 uM retinol supplement in maturation medium significantly stimulates embryo development, also improves the total cell number of blastocyst stage in pig.
Tak, Hyun-Min;Kim, Gyu-Tae;Kim, Eun-Jin;Mun, Yun-Ja;Choe, Chang-Yong;Son, Dong-Soo;Han, Jae-Hee;Kang, Da-Won
Journal of Embryo Transfer
/
v.24
no.1
/
pp.57-64
/
2009
This study was carried out to investigate expression of apoptosis-related differentially expressed gene (DEG) in ovaries of Korean cattle with follicular and luteal cysts and to identify the relationship between cyst and apoptosis using microarray, real-time PCR, TUNEL staining, and Western blot analysis. Microarray data showed that PIK3R2 and AKT1 were significantly up-regulated in follicular cyst, and TNF-RAF2, PRLR, FOXL2, STK4, and COL4A3 were up-regulated whereas INHA, CIDEB, BCL10, and FASLG were down-regulated in luteal cyst. Real-time PCR was performed to validate DEGs altered in luteal cyst. Of nine DEGs, four DEGs down-regulated in luteal cyst showed a positive corelation between microarray data and real-time PCR data. In this study, we focused on INHA, among many DEGs, which was highly down-regulated in both follicular and luteal cysts. Real-time PCR and micro array data showed that INHA was down-regulated by 12.3-fold and by 1.4-fold, respectively, in the bovine follicular cyst. TUNEL assay and Western blot analysis for ERK, JNK, p38, PI3K, and Akt, which were used to detect whether apoptosis is occurred, showed no significant changes in cystic ovaries (p>0.05). In the expression and activity of caspase-3, Bax, Bel-2, and Bel-xL, there was no significant changes between follicular cystic ovary and normal ovary. Rather, the expression levels of PI3K and p-Akt were decreased in follicular cystic ovary. These results suggest that deficiency of apoptosis in cystic ovary is associated with decreased expression of apoptotic effectors.
Journal of the Korean Society of Food Science and Nutrition
/
v.44
no.7
/
pp.975-982
/
2015
Piceatannol (trans-3,4,3',5'-trihydroxystilbene), a natural stilbene, is an analogue of resveratrol. In the present study, possible mechanisms by which piceatannol exerts its pro-apoptotic action in cultured human oral cancer YD-15 cells were investigated. To investigate whether or not piceatannol has effects on cancer cell viability, human oral YD-15 cells were treated with piceatannol (0, 50, and $100{\mu}M$). Piceatannol treatment ($100{\mu}M$) showed the strongest inhibition of cell proliferation and reduced cell viability in a dose-dependent manner. Chromatin condensation detected by DAPI staining significantly increased in a concentration-dependent manner, indicating apoptosis. Piceatannol treatment activated initiator Bax (pro-apoptotic) and cPARP in a concentration-dependent manner. Further, piceatannol induced down-regulation of Bcl-2 (anti-apoptotic). We also evaluated the activity of piceatannol against oral cavity cancer tumors in mice. Piceatannol-treated nude mice bearing YD-15 xenograft tumors exhibited significantly reduced tumor volume and weight due to the potent effect of piceatannol on tumor cell apoptosis, as determined by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assay. Immunohistochemistry staining showed elevated expression of cleaved-caspase-3 as well as reduced expression of Ki-67 in the piceatannol-treated group. Therefore, piceatannol can be developed as a cancer preventive medicine due to its growth inhibitory effects and induction of apoptosis in human oral cancer cells.
Human mesenchymal stem cells(hMSC), that have been reported to be present in bone marrow, adipose tissues, dermis, muscles and peripheral blood, have the potential to differentiate along different lineages including those forming bone, cartilage, fat, muscle and neuron. Therefore, hMSC are attractive candidates for cell and gene therapy. The optimal conditions for hMSC expansion require medium supplemented with fetal bovine serum(FBS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FBS proteins. Previously, we have shown that hADSC can be cultured in human serum(HS) during their isolation and expansion, and that they maintain their proliferative capacity and ability for multilineage differentiation and promote engraftment of peripheral blood-derived CD34 cells mobilized from bone marrow in NOD/SCID mice. In this study we determined whether hADSC grown in HS maintain surface markers expression similar with cells grown in FBS during culture expansion and compared gene expression profile by Affymetrix microarray. Flow cytometry analysis showed that HLA-DR, CD117, CD29 and CD44 expression in HS-cultured hADSC during culture expansion were similar with that in FBS-cultured cells. However, the gene expression profile in HS-cultured hADSC was significantly different from that in FBS-cultured cells. Therefore, these data indicated that HS-cultured hADSC should be used in vivo animal study of hADSC transplantation for direct extrapolation of preclinical data into clinical application.
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