• Journal of Korean Neurosurgical Society
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• 제29권9호
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• pp.1161-1170
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• 2000

Objective : The transverse hippocampal slice is one of the most commonly studied in vitro models of mammalian brain physiology. However, despite its broad usage, there has been no standardization of slice preparation techniques or recording condition. It is well known that variations in recording conditions can result in profound different effects to neuronal responses. Evoked field potentials, recorded extracellularly, were used to investigate the effects of variations in hippocampal slice preparation protocol on hypoxia responses of CA1 neurones. Material & Methods : Before hypoxic injury, hippocampal slices were incubated for 4 hours. During incubation period, the slices were placed in a incubation chamber($21^{\circ}C$) for recovery from preparation injury and then transferred to recording chamber($34^{\circ}C$) for more recovery and baseline electric recording with current stimulation(0.1Hz). Various time periods in incubation chamber and recording chamber were applied to each experimental group(group 1=60min : 180min, group 2=90min : 150min, group 3=180min : 60min, time in incubation chamber : time in recording chamber) before 10 min hypoxia produced by replacing 95% $O_2$+5% $CO_2$ mixed gas to 95% $N_2$+5% $CO_2$ gas. Calcium, Magnesium ions and several drugs effecting on glutamate receptor also were studied. Recoveries from hypoxic injury of hippocampal slices were estimated by percent recovery of population spike(PS). Statistic analysis of study were performed using paired t-test. Results : The percent recovery of PS after 10min hypoxia was considerably enhanced by increasing the period of current stimulation during incubation period before hypoxic injury. Temperature effect on the result of this experiment was also studied(group 4) but the result from this showed no statistic significance. Low magnesium ion concentration of artificial CSF(Mg-free aCSF) during incubation period enhanced the recovery of PS but low calcium (calcium-free) and high magnesium ion concentration(2mM) reduced it after hypoxic injury. L-glutamate($100{\mu}M$) and AP-5($50{\mu}M$) had no effect on the recovery of PS but CNQX($10{\mu}M$) in artificial CSF during incubation period markedly enhanced the recovery of PS. Co-treatment of AP-5($50{\mu}M$), CNQX($10{\mu}M$) and high magnesium concentration(2mM) enhanced recovery of PS in immediate following period of hypoxic injury but the effect of cotreatment after then decayed rapidly and lost statistic significance. Conclusions : Judging from above results, the condition of baseline recording is important in observing the recovery of population spike after hypoxia, and the time and the condition should be controled more strictly to obtain reliable results.

• Journal of Plant Biotechnology
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• 제40권4호
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• pp.224-230
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• 2013

본 연구 결과 질산은과 구연산 및 비타민 C의 처리는 레드플레임의 갈변화 방지에는 효과적이나, 레드플레임 인편의 재분화 및 생장에 있어 비타민 C와 질산은의 처리는 효과적 이지 못하였다. 반면 구연산의 경우 레드플레임의 재분화 및 생장에 도움을 주는 것으로 나타났으나 $150mg{\cdot}l^{-1}$이상의 고농도에서는 생육이 저해됨을 보여 $100mg{\cdot}l^{-1}$ 처리가 나리의 재분화 및 생장에 가장 효율적임을 확인하였다. 또한 절편체의 갈변화를 감소시키는 부분에서는 비타민 C $150mg{\cdot}l^{-1}$ 처리가 1.5%의 가장 낮은 갈변율을 나타내어 갈변화 현상 감소에 가장 적합한 처리구로 판단되었다. 본 실험결과는 향후 나리기내 식물체생산에 문제점으로 발생하는 갈변화 현상을 감소시키고 인편 재분화 및 생장을 촉진시켜 우량 나리 기내 묘의 대량증식체계 확립을 통한 우량종구묘 생산체계 확립에 기여하여 농가 소득 향상에 도움이 될 것으로 판단된다.

• Journal of Ginseng Research
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• 제23권1호
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• pp.13-20
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• 1999

사포닌 함량과 PD계 사포닌과 PT계 사포닌의 조성 비율이 상이한 사포닌 분획물 TS-1, TS-2, TS-3 즉 주종 ginsenoside의 총함량$(G-Rb_1,\;-Rb_2,\;-Rc,\;-Rd,\;G-Re,\;-Rf,\;-Rg_1)$은 각각 $41\%,40\%,62\%$이고, PD계 사포닌과 PT계 사포닌의 함유 조성비율(PD/PT)이 각각 1.55, 2.25, 2.61인 시료를 6년근 홍삼으로부터 추출 제조하여, 생체외 실험으로 음경해면체 절편에 대한 이완반응과 생체내 실험으로 사포닌분획물 투여에 따른 음경 해면체내압의 변화에 미치는 효과를 조사하였다. PHE에 의해 수축된 음경해면체 절편에 대한 이완 반응은 홍삼 사포닌은 각각 농도 의존적 이완작용을 보였으며, 음경 해면체평활근의 이완작용에 있어 각 분획물의 $ED_{50}$은 TS-1이 1.68, TS-2, 1.97, TS-3, 2.29 mg/ml서 TS-1이 가장 의의있게 낮았다. 이러한 홍삼 사포닌 분획물의 이완효과 기전은 음경 해면체평활근에 NO나 칼슘 및 칼륨 통로 등에 영향을 미쳐 복합적인 이완작용을 보이는 것으로 사료되었다. 흰쥐를 이용한 생체내 실험으로 홍삼 사포닌 투여에 따른 음경해면체내압의 변화 조사에서 각 사포닌 분획물은 농도의존적으로 해면체내압의 증가를 야기시켰다. 이들 해면체내압의 증가 정도는 TS-1이 가장 강하였고,TS-2,TS-3 순이었다. 이상의 결과로부터 홍삼의 사포닌 분획물은 음경해면체평활근의 이완작용과 내압의 증가에 영향을 미쳐 음경발기의 상승을 야기시키는 효과가 있으며 그 효과는 그 함유 조성에 따라 차별성이 있다는 것이 확인되었으므로 금후 주요 활성성분의 구명이 요망되고 있다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제43권2호
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• pp.77-82
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• 2017

Objectives: The aim of this study was to compare the mechanical resistance of four different osteosyntheses modeled in two different sagittal split ramus osteotomy (SSRO) designs and to determine the linear loading in a universal testing machine. Materials and Methods: An in vitro experiment was conducted with 40 polyurethane hemimandibles. The samples were divided into two groups based on osteotomy design; Group I, right angles between osteotomies and Group II, no right angles between osteotomies. In each group, the hemimandibles were distributed into four subgroups according to the osteosynthesis method, using one 4-hole 2.0 mm conventional or locking plate, with or without one bicortical screw with a length of 12.0 mm (hybrid technique). Each subgroup contained five samples and was subjected to a linear loading test in a universal testing machine. Results: The peak load and peak displacement were compared for statistical significance using PASW Statistics 18.0 (IBM Co., USA). In general, there was no difference between the peak load and peak displacement related to osteotomy design. However, when the subgroups were compared, the osteotomy without right angles offered higher mechanical resistance when one conventional or locking 2.0 mm plate was used. One locking plate with one bicortical screw showed higher mechanical resistance ($162.72{\pm}42.55N$), and these results were statistically significantly compared to one conventional plate with monocortical screws (P=0.016) and one locking plate with monocortical screws (P=0.012). The difference in peak displacement was not statistically significant based on osteotomy design or internal fixation system configuration. Conclusion: The placement of one bicortical screw in the distal region promoted better stabilization of SSRO. The osteotomy design did not influence the mechanical behavior of SSRO when the hybrid technique was applied.

• 한국동물학회지
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• 제38권4호
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• pp.523-530
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• 1995

돼지의 난소 조직내에 존재하는 심방이뇨 호르몬 수용체 분포를 알아 보기 위하여 자가방사법을 통해 125I로 표지한 rANP(1-28)의 특이적 결합 부위를 관찰하였다. 난소 조직 중 125I-rANP(1-28)의 강한 결합 부위는 난포의 과립막 세포층이었으며, 외난포막층에서도 125I-rANP(1-28)의 결합 부위가 관찰되었다. 그러나 내난포막층을 포함한 난소내의 다른 조직과 특히 황체에서는 125I-rANP(1-28)의 결합 부위가 나타나지 않았다. 난포의 과립막 세포층과 외난포막층에서의 이러한 125I-rANP(1-28)의 결합은 다량의 rANP(1-28)에 의하여 완전히 전위되었지만, 펩티드 호르몬인 angiotensin II 및 arginine vasopressin에 의해서는 과량의 농도에서도 전위되지 않아 125I-rANP(1-28)의 결합이 특이적임을 확인하였다. 또한 이러한 특이적 결하은 심방이뇨 호르몬의 생물학적 수용체 외에, 다른 기능을 담당하는 clearance 수용체의 특이적 ligand인 C-ANF에 의해서도 전이되었다. 이상의 결과는 돼지의 난소에 있어서 난포의 과립막 세포층 및 외난포막에 심방이뇨 호르몬의 생물학적 또는 coearance 수용체가 존재함을 보여주며, 이는 심방이뇨 호르몬의 수용체가 난자의 성숙에 관련된 난포의 발달과정에 관여할 수 있음을 시사한다.

• Clinical and Experimental Reproductive Medicine
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• 제45권3호
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• pp.129-134
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• 2018

Objective: In frozen and thawed embryos, the zona pellucida (ZP) can be damaged due to hardening. Laser-assisted hatching (LAH) of embryos can increase the pregnancy rate. This study compared thinning and drilling of the ZP before frozen embryo transfer (FET). Methods: Patients were randomly allocated into two groups for LAH using thinning or drilling on day 2 after thawing. Twenty-five percent of the ZP circumference and 50% of the ZP thickness was removed in the thinning group, and a hole $40{\mu}m$ in diameter was made in the drilling group. Results: A total of 171 in vitro fertilization/intracytoplasmic sperm injection FET cycles, including 85 cycles with drilling LAH and 86 cycles with thinning LAH, were carried out. The thinning group had a similar ${\beta}$-human chorionic gonadotropin-positive rate (38.4% vs. 29.4%), implantation rate (16.5% vs. 14.4%), clinical pregnancy rate (36.0% vs. 25.9%), miscarriage rate (5.8% vs. 2.4%), ongoing pregnancy rate (30.2% vs. 23.5%), and multiple pregnancy rate (7.0% vs. 10.6%) to the drilling LAH group. There were no significant differences in pregnancy outcomes between subgroups defined based on age (older or younger than 35 years) or ZP thickness (greater or less than $17{\mu}m$) according to the LAH method. Conclusion: The present study demonstrated that partial ZP thinning or drilling resulted in similar outcomes in implantation and pregnancy rates using thawed embryos, irrespective of women's age or ZP thickness.

• Journal of Yeungnam Medical Science
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• 제15권2호
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• pp.286-296
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• 1998

비중격 만곡증 환자의 비내수술시 채취한 정상 하비갑개 점막조직으로 부터 상피세포만을 분리세포의 단층배양법(monolayer culture of dissociated cells)으로 배양하여 비점막 상피세포 배양법을 정립하고 또한 배양된 세포가 상피세포임을 동정하기 위하여 간접 면역형광항체법으로 상피세포 특유의 cytokeratin을 확인하였고 투과전자현미경으로 상피세포만이 가지고 있는 교소체(desmosome) 및 장세사(tonofilament) 등을 확인하였으며 6회까지 계대배양을 할 수 있었다. 향후 본 연구에서 배양된 비점막 상피세포를 이용하여 알레르기성 비염 등과 같은 비점막 염증성 질환의 병태생리에서 상피세포의 역할에 대한 연구와 호산구등 염증 세포의 침윤과 상피세포의 손상에 접착분자 및 cytokine의 상호작용에 관한 연구가 계속될 수 있을 것으로 기대한다.

• Clinical and Experimental Reproductive Medicine
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• 제24권2호
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• pp.153-165
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• 1997

In male reproducible health, fertility and IVF (in-vitro fertilization), semen analysis has been most important. Semen analysis can be divided into concentration, motional and morphological analysis of sperm. The existing method which was developed earlier to analyze semen concentrated on the sperm motility analysis. To provide more useful and precise solutions for clinical problems such as infertility, semen analysis must include sperm morphological analysis. But the traditional tools for semen analysis are subjective, imprecise, inaccurate, difficult to standardize, and difficult to reproduce. Therefore, with the help of development of microcomputers and image processing techniques, we developed a new sperm morphology analyzer to overcome these problems. In this study the agreement on percent normal morphology was studied between different observers and a computerized sperm morphology analyzer on a slide-by-slide basis using strict criteria. Slides from 30 different patients from the SNUH andrology laboratory were selected randomly. Microscopic fields and sperm cells were chosen randomly and percent normal morphology was recorded. The ability of sperm morphology analyzer to repeat the same reading for normal and abnormal cells was studied. The results showed that there was no significant bias between two experienced observers. The limits of agreement were 4.1%${\sim}$-3.8%. The Pearson correlation coefficient between readers was 0.79. Between the manual and sperm morphology analyzer, the same findings were reported. In this experiments the slides were stained by two different methods, PAP and Diff-Quik staining methods. The limits of agreement were 7.2%${\sim}$-5.7% and 6.0%${\sim}$-6.3%, respectively. The Pearson correlation coefficients ware 0.76 and 0.91, respectively. The limits of agreement was tighter below 20% normal forms. In the experiments of repeatability, 52 cells stained by PAP and Diff-Quik staining methods were analyzed three times in succession. Estimating pairwise agreement, the kappa statistic for the pairs were 0.76, 0.81, 0.86, and 0.75, 0.88, 0.88 respectively. In this study it was shown that there was good agreement between manual and computerized assessment of normal and abnormal cells. The repeatability and agreement per slide of computerized sperm morphology analyzer was excellent. The computer's ability to classify normal morphology per slide is promising. Based on results obtained, this system can be of clinical value both in andrology laboratories and IVF units.

• Archives of Plastic Surgery
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• 제37권5호
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• pp.531-534
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• 2010

Purpose: Lipobean$^{(R)}$s, widely used in lipodissolving techniques, contain phosphatidylcholine and sodium deoxycholate as its main substances. They have been approved only as medication for liver disease by the FDA. However, they have been used under various clinical settings without exact knowledge of its action mechanism. The authors designed an in vitro study to analyze the effects of different concentrations of phosphatidylcholine and sodium deoxycholate on adipocytes and other types of cells. Methods: Human adipose-derived stem cell were cultured and induced to differentiate into adipocytes. Fibroblasts extracted from human inferior turbinate tissue, and MC3T3-E1 osteoblast lines were cultured. Phosphatidylcholine solution dissolved with ethanol was applied to the culture medium at differing concentrations (1, 4, 7, 10 mg/mL). The sodium deoxycholate solution dissolved in DMSO applied to the medium at differing concentrations (0.07, 0.1. 0.4. 0.7 mg/mL). Cells were dispersed at a concentration of $5{\times}10^3$ cells/well in 24 well plates, and surviving cells were calculated 1 day after the application using a CCK-8 kit. Results: The number of surviving cells of adipocytes, fibroblasts and osteoblasts decreased as the concentration of sodium deoxycholate increased. However, all types of cells that had been processed in a phosphatidylcholine showed a cell survival rate of over 70% at all concentrations. Conclusion: This study shows that sodium deoxycholate is the more major factor in destroying adipocytes, and it is also toxic to the other cells. Therefore, we conclude that care must be taken when using Lipobean$^{(R)}$s as a method of reducing adipose tissue, for its toxicity may destroy other nontarget cells existing in the subcutaneous tissue layer.

• Journal of Periodontal and Implant Science
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• 제34권1호
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• pp.49-59
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• 2004

The goal of periodontal treatment is not only to arrest the progression of the disease but also to promote the functional, esthetic regeneration of the periodontium. Flap operation, bone graft, guided tissue regeneration, growth factors and bone morphogenetic protein have been used for this purpose. Among these techniques of regeneration, alloplastic graft, especially calcium phosphate is getting more attention recently. The purpose of this study was to evaluate the effects of calcium phosphate glass on mouse calvarial cell in vitro. The toxicity of calcium phosphate glass was measured using MTT assay, the synthesis of collagen was measured using collagen assay, and ALP activity was measured. The experimental groups were cultured with calcium phosphate glass(both AQ-, and HT-CPG) in concentration of 0.01, 0.02, 0.1, 0.2g/ml. The results are as follows 1. In concentrations not exceeding 0.02g/ml, both the groups(AQ-CPG, HT-CPG) didn't show any toxicity on mouse calvarial cell(p<0.05). 2. In both the experimental groups are the concentration of 0.02g/ml, collagen expressions were significantly up-regulated (p<0.05). 3. In both the experimental groups are the concentration of 0.02g/ml, ALP activity was not significantly up-regulated, but ALP activity in both experimental groups were greater than control group(p<0.05). The results suggested that the use of calcium phosphate glass may promotes periodontal regeneration. Ongoing studies are necessary in order to determine their regeneration effects.