• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.103-108
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• 1996

Through the previous studies(I,II), it was observed that human follicular fluid(HFF) was more effective than human fetal cord serum(HFCS) on promoting meiotic resumption of oocytes and improving embryonic development of mouse in vitro. On the basis of these results, we have gradually exchanged HFCS with HFF as protein supplement in human ART. This study was performed to investigate the efficiency of HFF on improving the pregnancy rate in ART. Oocytes were retrieved transvaginally from patients treated with pituitary suppression with GnRH-agonist and ovarian stimulation with human menopausal gonadotro-pin(HMG) and pure follicle stimulating hormone(FSH). Aspirated oocytes were rinsed and cultured in TCM-199 containing HFF, and the concentrations of HFF were adjusted to 10, 20, and 30% according to the use for insemination, embryo growth and embryo transfer, respectively. As possible as, we tried to do embryo transfer into fallopian tube to mimic the coincidence of the cell stage with the place of sojourn in vivo, so we performed various ART programs(IVF & ET; in vitro fertilization, ZIFT; zygote intra fallopian-tube transfer, ZIFT & ET) according to the tubal conditions of patients. On the while, intra cytoplasmic sperm injection(ICSI) was used to assist IVF of the patients who had shown poor standard IVF results by immunological or severe male factor. Of the 255 cycles of ART programs using HFF as protein supplement, 118 cycles were turn out to be succeeded in pregnancy(46.2%, per cycle, p<0.05), while 21 pregnancies were achieved in the 69 cycles using HFCS(30.4%). The 255 cycles using HFF were subdivided into cycles with the type of ART programs, and each pregnancy rate of the ART programs were 44.7% (IVF & ET, 76/170 cycles), 53.4%(ZIFT, 31/58 cycles) and 40.7% (ZIFT & ET, 11/27 cycles), respectively. In the 61 ICSI cycles using HFF, 28 cycles succeed in pregnancy(45.9%), while 7 pregnancies were obtained in the 17 ICSI cycles using HFCS. Also the ongoing pregnancy rate in the group using HFF(78.8%, 93/118 cycles) was higher than that in the group using HFCS(61.9%). Therefore, we found that the use of HFF as protein supplement was more suitable and effective than the use of HFCS to improve the pregnancy rate in ART.

• Neonatal Medicine
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• 제18권2호
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• pp.280-287
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• 2011

목적: 미숙아에서 위액 내 Ureaplasma urealyticum 의 검출이 미숙아의 특징과 임상적 예후에 어떤 영향을 주는지를 알아보고자 하였다. 방법: 미숙아를 대상으로 출생 후 30분 이내에 첫 위액은 채취하였고, 위액 내 U. urealyticum 이 검출되는지를 확인하기 위해 24시간 이내에 배양을 실시하였다. 그리고 U. urealyticum 이 검출된 군을 양성군으로, 검출되지 않은 군을 음성군으로 나누었다. 결과: 91명의 미숙아 중에서 17명(19%)에서 위액 내 U. urealyticum 이 검출되었다. U. urealyticum 양성군에서 음성군보다 통계학적으로 30주 미만의 미숙아 수가 더 많았고(P=0.02), 1분과 5분 아프가 점수가 더 높았으며(P=0.017과 P=0.048), 그리고 질식 분만이 더 많았다(P=0.000). 두 군간에 기관지폐이형성증의 발생률은 차이가 없었지만, 이전에 신생아호흡 곤란증 진단 없이 기관지폐이형성증의 발생한 빈도는 음성군(1%)보다 양성군 (11%)에서 의미 있게 높았다(P=0.03). 결론: 위액 내 U. urealyticum 검출은 임신 기간 30주 미만 미숙아에서 높게 관찰되고, 미숙아에서 선행되는 신생아호흡곤란증후군이 없이도 발생할 수 있는 기관지폐이형성증을 예측하는데 도움이 된다.

• Archives of Plastic Surgery
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• 제37권3호
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• pp.227-232
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• 2010

Purpose: Continuous irrigation method is an important step in managing wound infection. V.A.C. devices have been used in intractable wounds for reducing discharge, improving local blood flow, and promoting healthy granulation tissue. We expect synergistic effects of reduced infection and more satisfactory, accelerated wound healing when using both methods simultaneously. This study evaluated continuous irrigation combined with V.A.C. appliance for treatment of infected chronic wounds. Methods: We reviewed data from 17 patients with infected intractable chronic wounds. V.A.C. device (Group A) was used in 9 patients, and V.A.C. with antibiotics irrigation (Group B) was used in 8 patients. We placed Mepitel$^{(R)}$ on the surface of wound and placed an irrigation and aspiration tube on each side. A sponge was placed on the Mepitel$^{(R)}$ and covered with film dressing. The wound was irrigated continuously with mixed antibiotics solution at the speed of 200 cc/hr and aspirated through the wall suction at the pressure of -125 mmHg. V.A.C. applied time, wound culture and wound size were compared between the two groups. Results: No complication were seen in two groups. Compared with Group A, in the Group B, V.A.C. applied time was shortened from 32.7 days to 25.6 days and showed efficacy in the reduction rate of wound size. No statistical differences were shown in bacterial reversion. Conclusion: V.A.C. appliance with continuous irrigation is an effective new method of managing infected chronic wounds and useful to reduce treatment duration and decrease wound size. Moreover it could be applied more widely to infected wound.

• 대한한의학회지
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• 제16권1호통권29호
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• pp.209-226
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• 1995

In order to investigate the protective effects of Jiyusaenggan-Tang by F-I(Jiyusaenggan-Tang aspirated by distilled water), F-II(Non-polysaccharides in Jiyusaenggan-Tang) and F-III(Polysaccharides in Jiyusaenggan-Tang), experimental studies were performed in mice with liver injury induced by $CCI_4$, and d-galactosamine. The results were summarized as follows : 1. The increases of the serum GOT activities in mice induced by $CCI_4$ were inhibited significantly by the administration of highly concentrated FIII. The increases of the serum GPT, LDH, and ALP activities in mice induced by $CCI_4$ were inhibited significantly by the administration of highly concentrated FI and FIII. 2. The increases of the serum total-cholesterol and triglyceride activities in mice induced by $CCI_4$ were inhibited significantly by the administration of highly concentrated FI, FII and FIII. 3. On liver injury induced by the $CCI_4$, the activities of GOT, GPT, ALP, LDH were inhibited by highly concentrated FIII, F-I, F-II, and F-I, F-II, F-III on total cholesterol level and F-III, F-I, F-II on triglyceride level in order of their efficacies. And F-III is considered to be the most effective component. 4. The increases of the serum GOT, GPT activities in mice induced by d-galactosamine were inhibited significantly by the administration of FI and FIII. The increases of the serum LDH, ALP activities in mice induced by d-galactosamine were inhibited significantly by the administration of FI and FIII. 5. The increases of the serum Total-cholesterol, Triglyceride activities in mice induced by d-galactosamine were inhibited significantly by the administration of FI, FII and FIII. 6. The increases of the serum GOT, GPT activities in mice induced by d-galactosamine were inhibited significantly in F-III administrated group and then F-I in order of efficacies. FII does not showed significant effect, on inhibiting the activities of serum GOT and GPT levels. The activity of ALP was inhibited by F-III, F-I, F-II, LDH by F-III, F-I in order of efficacies, and F-II did not showed significant effect on LDH activity. The triglyceride level was decreased significantly by F-III, F-II, F-I and the total cholesterol level by F-II, F-I, F-III in order of their efficacies. Judging from the above results, it is considered that Jiyusaenggan-Tang has protective effect against liver injury, and that Polysaccharides in Jiyusaenggan-Tang is more effective.

• 미생물학회지
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• 제39권1호
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• pp.36-39
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• 2003

본 연구에서는 16S rDNA복합중합효소연쇄반응을 이용하여 중이 삼출액에서 미생물병인원에 대한 특성을 알아보았다. 중이염환자의 중이 삼출액에서의 미생물 병인원은 주로 streptococcus pneumoniae, Haemophilus influenzae와 Moraxella catarrhalis이다. 26명의 환자로부터 39개의 중이염의 삼출액을 얻었고, 중이 삼출액에서 DNA를 추출하였다. PCR은 16S rDNA의 C4 region에서 21 base pair의 common primer와 각각 bacterium specific primers [(i) Haemophilus-specific primer (ii) Moraxella-specific primer and (iii) Streptococcus-specific primer]를 이용하여 수행하였다. 39 개의 중이염의 삼출액 시료 중에서, H. influenzae가 24 개(61.5%) 검출되었고, M. catarrhalis는 10 개(25.6%), S.pneumoniae는 3개 (7.7%)가 검출되었다. 16s rDNA 복합중합효소연쇄 반응 진단 결과,11 개(28%)의 중이삼출액 시료에서 음성을 나타내었다. 중이염의 중복감염은 9개의 중이 삼출액시료에서 관찰되었고, 이들은 모두 H.influenzae 와 M. catarrhalis 에 의한 중복감염이었다. 본 연구에서 저자 등은 165 rDNA 복합중합효소연쇄반응이 병원성 미생물을 빠르게 진단하고, 중이삼출액의 미생물 병인론을 추적할 수 있는 좋은 방법으로 제시하는 바이다.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.67-67
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• 2003

Early development of porcine oocytes fertilized in vitro was examined in different culture conditions. Porcine ovaries were collected from local slaughter-house. Cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cystein, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for 42~44 hrs. The frozen-thawed spermatozoa were washed by centrifigation 2 times at 1, 500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, 1$\mu\textrm{g}$/ml streptomycin and 1ng/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to 2.5$\times$10$^{6}$ cells/ml motile sperm during fertilization in vitro. At 8hrs h after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine and 4 mg/ml BSA and cultured for 7 days. In first experiment, the mean numbers of oocytes collected from 20 ovaries were 674.4 oocytes, and 4.1(27.6), 12.5(84.0), 25.4(171.6) and 57.9%(390.8) for A, B, C and D grade in morphological classification. In the second experiment, when culture medium was supplemented with various concentrations of EGF, the proportions of oocytes cleaved were 56.9, 55.7, 61.9 and 54.7% for 0, 5, 10 and 20ng/ml EGF. The higher proportions(15.1%) of oocytes developed to morular stage were obtained at concentration of 10ng/ml than 0 and 5ng/ml EGF (P<0.05). However, the proportions of embryos developed to blastocyst stage were not significantly different among concentrations of EGF. In another experiment, when the medium supplemented with catalase was used, the proportions of oocytes cleaved were higher in the concentration of 0 unit (56.5%, 61/108) than 100 and 1, 000 unit/ml of catalase (P<0.05). Although the developmental capacity of embryos was improved by medium with 0 unit/ml compared with 100, 500 and 1, 000 units/ml of catalase in oocytes developed to morula and blastocyst stages, were not significantly different among concentrations of catalase.

• 한국수정란이식학회지
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• 제15권3호
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• pp.231-236
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• 2000

This study was conducted to examine the effects of exogenous gonadotropins (PMSG+hCG) and an antioxidant (cysteine) on in vitro maturation of bovine follicular oocytes. Cumulus-oocyte complexes (COCs) aspirated from 2 to 5 mm ovarian follicles were cultured for 22 to 24 hours in a modified bovine embryo culture medium (mBECM) supplemented with 3 mg/mL bovine serum albumin, to which PMSG (10 IU/mL) + hCG (10 IU/mL) and/or cysteine (0.6 mM) were added. When examined the expansion of cumulus ce1ls at the end of maturation culture, greater (p<0.05) expansion was found after addition of PMSG+hCG (79 to 96%) to mBECM than after no addition (0%), regardless of the presence or absence of cysteine in the medium. The addition of cysteine did not stimulate cumulus expansion, but a high proportion (92%) of expansion was achieved when COCs were cultured after the addition of PMSG+hCG and cysteine to the medium. No difference in the proportion of oocytes underwent germinal vesicle breakdown (initiation of maturation) was found after the addition of PMSG+hCG and/or cysteine to mBECM. However, nuclear maturation (development to the metaphase-II stage) of oocytes was significantly stimulated by the combined addition of PMSG+hCG and cysteine, compared with no addition. In conclusion, both exogenous gonadotropins and an antioxidant are important for nuclear maturation of bovine immature oocytes and these factors have a cell-specific stimulatory action.

• Reproductive and Developmental Biology
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• 제29권2호
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• pp.127-131
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• 2005

The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from $2\~6mm$ follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, $0.5{\mu}/ml$ FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at $39^{\circ}C$ in a humidified atmosphere of $5\%$ $CO_2$ in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with $1\%$ orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm $(1\times10^5\; cells/ml)$ in mTBM with $0.3\%$ BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with $0.4\%$ BSA. At 6hr of culture, the embryos were fixed in $3.7\%$ formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and $\geq3$ PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group $(67\%)$, but it did not differ among the all added groups $(86\%,\;85\%,\;79\%\;and\;81\%$, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups $(25\%,\;30\%,\;33\%,\;29\%\;and\;29\%$, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased ($66\%\;vs\;. 58\%,\;54\%,\;52\%\;and\;55\%$, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

• Reproductive and Developmental Biology
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• 제30권2호
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• pp.119-124
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• 2006

The present study compared the developmental potential of cloned porcine embryos with mesenchymal stem cells (MSCs), fetal fibroblasts (FFs) and cumulus cells (CCs) by assessing the cleavage and blastocyst rate, total cell number, inner cell mass (ICM) ratio and apoptosis. MSCs were isolated by ficoll gradients from femur of -6 month old female pig, and maintained for primary cultures. FFs from a female fetus at ${\sim}30$ day of gestation were established, and CCs were obtained from cumulus oocyte complexes (COCs) aspirated from $3{\sim}6$ mm follicles in diameter. Donor cells at $3{\sim}4$ passage were employed for nuclear transfer (NT). COCs were matured and fertilized in vitro(IVF) as control. Cleavage rate was significantly (P<0.05) higher in IVF than in NT embryos with MSCs, FFs and CCs ($82.7{\pm}8.9%\;vs\;70.6{\pm}5.4,\;68.7{\pm}5.1\;and\;63.4{\pm}5.6%$, respectively). However, blastocyst rates in IVF and NT embryos derived from MSCs ($24.5{\pm}2.8\;and\;20.4{\pm}8.3%$) did not differ, but were significantly (P<0.05) higher than NT derived from FFs and CCs ($10.6{\pm}2.7\;and\;9.8{\pm}2.1%$). Total cell number and the ratio of ICM to total cells among blastocysts cloned from MSCs ($35.4{\pm}5.2\;and\;0.40{\pm}0.09%$, respectively) were significantly (P<0.05) higher than those from FFs and CCs ($24.9{\pm}6.2%\;vs\;0.19{\pm}0.16,\;23.6{\pm}5.5\;and\;0.17{\pm}0.16%$, respectively). Proportions of TUNEL positive cells in NT embryos from FFs and CCs ($6.9{\pm}1.5\;and\;7.4{\pm}1.7%$, respectively) were significantly (P<0.05) higher than in MSCs ($4.8{\pm}1.4%$) and IVF ($2.3{\pm}0.9%$). The results demonstrate that MSCs have a greater potential as donor cells than FFs and CCs in achieving enhanced production of cloned porcine embryos.

• Clinical and Experimental Reproductive Medicine
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• 제29권3호
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• pp.201-207
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• 2002

Objective: To observe the capability of fertilization and embryo development including blastocyst formation of the oocytes in simple media after thawing of the cryopreserved cumulus-free mouse oocytes by vitrification method. Methods: Oocytes were collected from 5 to 6 weeks old ICR female mice, and were denuded from the cumulus cells by 0.1% hyaluronidase. Recovered mature oocytes in study group were cryopreserved by vitrification method using EM grid for $5{\sim}7$ days. In brief, oocytes were exposed in dPBS containing 1.5 M EG and 5.5 M EG+1 M sucrose for 2.5 minutes and 20 seconds each, and then executed vitrification by plunging in LN2 after loading on EM grid. Thawing treated by exposure of 1, 0.5, 0.25 and 0.125 M sucrose solution for 2.5 minutes each in order and used for experiments. Spermatozoa aspirated form the epididymis of 12 weeks old ICR male mice were used for insemination after capacitation. T6 media containing 0.4% BSA were used for fertilization and development. Results: Survival and fertilization rates after thawing were 76.9% and 79.6% respectively. Fertilization rate was lower (p<0.005) than that of control group (92.9%). There was no difference in embryo developmental rates from 2-cell to morula, however, the blastocyst formation rate and mean cell numbers of blastocysts in study group (63.3%, $58.9{\pm}9.2$) were lower compared with those of control group (76.1%, $63.5{\pm}8.9$). Conclusion: Vitrification is an effective method for mouse mature oocyte cryopreservation with high survival and fertilization rate after thawing. And in simple media, fertilization rates and embryo development of frozen-thawed mouse oocytes are satisfactory.