• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.63-63
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• 2003

This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.7
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• pp.1212-1217
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• 2004

The aim of this study was to investigate Quality characteristics of Pleurospermum kamtschaticum to increase the value of functional food resources. To examine Quality characteristics of Pleurospermum kamtschaticum, various factors such as color, texture, fiber, minerals, tannin, crude proteins, crude lipids and sensory quality, were determined using physicochemical methods. The contents of dietary fiber were 0.57％ and 0.54％ in each 100 g of leaf and stem parts (freeze drying base). In mineral content, potassium was the highest value in both of leaf and stem parts (freeze drying base). The contents of tannin were 108.1 $\mu$g/mL and 20.9 $\mu$g/mL in leaf and stem parts (fresh base) respectively. Blanching stems showed significant difference in hardness from freeze-drying leaves. The antioxidative activity in ethanol extracts of Pleurospermum kamtschaticum was investigated using peroxide value and free radical scavenging activity. Free radical scavenging activity in ethanol extracts of Pleurospermum kamtschaticum was 43.5％ in a 100 $\mu$g/mL level, and the antioxidative index was 1.09 in a 500 $\mu$g/mL level. For functional food using Pleurospermum kamtschaticum the contents of crude proteins and lipids were about 9％ and 26％, respectively. The results from sensory evaluation of each treatment older age showed a better acceptability.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.145-151
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• 2002

The aim of these experiments was to investigate in vitro nuclear maturation of canine oocyte collected from various stages of estrus cycles, Ovaries were obtained from 1 to 4 year-old mongrel bitch and minced for oocyte collection in phosphate buffered saline with 100 iu penicillin-G $m\ell$／sup -1／, 50 $\mu\textrm{g}$ streptomycin sulphate $m\ell$／sup -1／ and 0.1％ (w／v) polyvinyl alcohol. Cumulus-oocyte-complexes (COCs) were washed in HEPES buffered tissue culture medium (TCM)199 and in vitro matured in TCM-199 culture medium supplemented with sodium pyruvate 0.028mg/$m\ell$, L-glutamine 0.146mg／$m\ell$, penicillin G 10,000 IU／$m\ell$, streptomycin 0.031mg／$m\ell$ and 10％ (v／v) fatal calf serum. COCs were in vitro matured for 48～72 hrs at 39$^{\circ}C$ in humidified 5％ $CO_2$ in air atmosphere. In vitro matured oocytes were remove the cumulus cells using 0.2％ (v／v) hyaluronidase. After denuding, oocyte were placed in acetic acid : methanol : chlorform solusion (3:6 : 1.5 v／v) for 30 sec and acetic acid: ethanol(1:3 v／v) for 48hrs fixation. Nuclear maturation was classified to GV, GVBD, MI, MII and degenerate oocyte under microscopy after 1％ aceto-orcein stain. In vitro maturation rates at 48hrs were not significantly difference among the oocytes collected from different stage of estrus at 15.9％, 16.3％, 23.7％ and 18.2％ for anestrus, proestrus, estrus and diestrus. However, the oocytes maturation(36.6％) of collected from estrus ovaries were significantly different from oocytes derived from proesturs, diestrus and anestrus ovaries(30.8％, 17.5％ and 22.1％; p＜0.05). The overall in vitro maturation rates were significantly higher (p＜0.05) in 72hrs culture than 48hrs culture system. In summary, there was a tendency for higher in vitro maturation rates with the oocyte collected from estrus ovary than other stages of estrus. Also, for nuclear maturation, in vitro culture of oocyte for 72hrs was better than 48hrs culture.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.111-117
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• 1996

This experiment was designed to evaluate the effect of transforming growth factor-$\beta$ (TGF-$\beta$) and insulin-like growth factor-I (IGF-I) in bovine oocyte maturation in the presence or absence of serum on subsequent fertilization and embryo development. In addition, various concent rations of these growth factors were evaluated for the ability to promote development of eight-cell stage embryos to the blastocyst stage. Cumulus-oocyte complexes were recovered from 2 to 6 mm follicles obtained from slaughterhouse ovaries and cultured at 38.5$^{\circ}C$ for 24 hours in TCM-199 (HEPES Modification) with or with out 20 % fetal bovine serum (FBS) to which the following growth factors were added TGF$\beta$ IGF-l or TGF $\beta$ + IGF-I, all at 10 ng/ml each. The matured oocytes were fertilized in IVF-TL medium with frozen-thawed semen at a concentration of 1 ${\times}$ 10$^6$ cells/ml of fertilization medium following Percoll separation. After 24 hours of sperm-egg incubation, the embryos were transferred to CZB medium without glucose for 48 hours and then cultured in TCM-199 with 20% fetal bovine serum (FBS) for 96 hours. The addition of growth factors to IVM medium in the presence of serum had no effect on cleavage and subsequent embryo devlopment to blastocyst. In the absence of serum, TGF- improved cleavage and development to blastocyst compared to control's(p<0.05) and no synergistic effeet of IGF-I + TGF-$\beta$ was observed. In the second experiment, eight-cell embryos obtained by in vitro maturation (IVM) in TCM-199 + 20% FBS without growth facrors and in vitro fertil-ization (IVF) were cultured in the in vitro cuiture (IVC) medium supplemented with 5, 10 ng/ml TGF-$\beta$ or 5, 10, 50, 100 ng/ml IGF-I. Cleavage rate and development to the blastocyst stage was observed during seven days of incubation. The supplementation of 10 ng/ml TGF-$\beta$ to lVC medium for eight-cell embryos improved development to blastocyst (p<0.05) compared to control. In conclusion, these data indicate that the supplementation of growth factors to IVM medium in the presence of serum does not influence cleavage and subsequent embryo development. However, significantly more oocytes matured in serum-free TCM-199 and eight-cell embryos cultured in lVC medium developed to blastocyst with supplementation of 10ng/ml TGF-$\beta$.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.191-199
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• 2004

This study was conducted to examine in vitro development of porcine embryos constructed by the microinjection of cultured fetal fibroblast cells into porcine oocytes matured in vitro. Single fetal donor cells were deposited into the perivitelline space of enucleated oocytes, followed by electrical fusion and activation. Activated embryos were cultured in NCSU-23 medium supplemented with 5％ FBS, at 38.5$^{\circ}C$ for 6 to 8 days in 5％ $CO_2$ and air. In experiment 1, fusion rates of nuclear transfer embryos did not differ for fetal fibroblast cells incubated in 5％ FBS + NCSU-23 or 5％ FBS + TL Heaps medium, nor did fusion rates of donor cells differ between 1-8 hr incubation durations. Fusion rates for the four treatment subclasses ranged from 72.1％ to 78.0％. In experiment 2, Pre-synchronization in medium containing 0.1 $\mu\textrm{g}$/m Hoechst 33342 an increase from 0 and 8 versus 15 h culture an increased percentage of porcine fibroblast cells in G2/M at the end of the synchronization period (12.4％, 17.5％ and 47.6％). Neither an increase in the concentration of H 33342 (0.2-1.6 $\mu\textrm{g}$/$m\ell$) nor a longer exposure time (12h, 18h and 24h) increased the proportion of porcine G2/M fibroblasts. In experiment 3, fusion rates did not differ significantly far nuclear transfer embryos constructed using donor cells cultured in 5％ FBS + NCSU-23 medium for 1-2, 6-8 or 12-14 days (60.0％, 73.3％ and 62.5％), respectively. The cleavage rate for nuclear transplant embryos using fetal fibroblast cells cultured for 1-2 days was 44.0％, significantly less than 56.7％ and 50.0％. for 6-8 or 12-14 days duration of culture, respectively. In experiment 4, the proportions of nuclear transfer embryos that developed to the $\geq$2 cell and to the blastocyst stage were not affected significantly by culture medium (5％ FBS + NCSU-23 or 5％ FBS + TL-Heaps) or by $O_2$ concentration of the culture (5％ vs 10％). Rates of development to the $\geq$2 cell stage ranged from 65.9％ to 70.1％, and development rates to the blastocyst stage ranged from 9.8％ to 12.5％ for the four treatment subclasses. Developmental rate was highest for embryos cultured in 5％ FBS + NCSU-23 under a gas atmosphere of 5％ $O_2$ in air.

• The Journal of the Petrological Society of Korea
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• v.12 no.1
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• pp.16-31
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• 2003

The granites in the southern Gimcheon area can be divided into two parts, marginal hornblende biotite granodiorite (Mgd) and central biotite granodiorite to granite (Cgd). Mgd and Cgd are gray in color and display gradational contact relations and are mainly composed of coarse-grained and medium-grained rocks, respectively. Mgd has more frequent and larger mafic enclaves than Cgd, and the two granites partly show parallel foliation at thire contact with gneisses. From representative samples of the granites, K-Ar biotite ages of 197∼207 Ma were obtained. Considering the blocking temperature of biotite, it is suggested that the emplacement age of the granitic magma was probably late Triassic. The anorthite contents of plagioclases in Mgd display less variation than those of Cgd, indicating that Mgd crystallized within a narrow range of temperatures. In the Al$\_$total/-Mg diagram, the biotites from the granites plot within the subalkaline field, and the smooth slope indicates differentiation from a single magma. All amphiboles from the granites belong to magnesio-hornblende. The linear trends of major oxides, AFM and Ba-Sr-Rb indicate that Mgd and Cgd were fractionally differentiated from a single granitic magma body crystallizing from the margin inwards. The relations of modal (Qz+Af) vs. Op, K$_2$O vs. Na$_2$O, Fe$_2$ $O_3$ vs. FeO, Fe$\^$+3/(Fe$\^$+3/+Fe$\^$+2/) and K/Rb vs. Rb/Sr show that they belong to I-type and magnetite-series granitic rocks developed by the progressive melting products of fixed sources. REE data, normalized to chondrite value, have trends of enriched LREE and depleted HREE together with weakly negative Eu anomalies.

• Journal of Preventive Medicine and Public Health
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• v.25 no.1 s.37
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• pp.13-25
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• 1992

This study was conducted to investigate the mechanism of protective effect by sodium selenite in methylmercuric chloride neurotoxicity, increasing intracellular $Ca^{2+}$concentration of the neuron. Methylmercuric chloride of 3mg/kg of body weight was administered simultaneously with sodium selenite of 5mg/kg and pretreatment of sodium selenite via intraperitoneal injection to rats. Also, effect of methylmercuric chloride($25{\mu}M,\;50{\mu}M,\;100{\mu}M$) and sodium selenite($200{\mu}M$) on free intrasynaptosomal $Ca^{2+}$ concentration were studied using the fluorescent $Ca^{2+}$ indicator fura -2 in vitro. After the treatment, at 6, 24, and 48 hours later, mercury in the cerebral cortex, liver and kidney tissues, succlnic dehydrogenase activities, adenosin-5'-triphosphate concentration, acetylcholinesterase activities, and intracellular $Ca^{2+}$ concentration in the cerebral cortex were determined in vivo. Cerebral synaptosomes of rats were incubated with methylmercuric chloride and sodium selenite in Hepes buffer for 10 minutes and free intrasynaptosomal $Ca^{2+}$ concentration were measured with fura-2 in vitro. The results were summarized as follows ; 1. The combined administration of $CH_3HgCl$ and $Na_2SeO_3$ and pretreatment of $Na_2SeO_3$ according to time significantly more increased in the cerebral cortex and decreased in the liver, kidney mercury concentrations compared to the administration of $CH_3HgCl$ only. 2. The combined administration of $CH_3HgCl$ and $Na_2SeO_3$ and pretreatment of $Na_2SeO_3$ increased more succinic dehydrogenase and acetylcholinesterase activities compared to the administration of $CH_3HgCl$ only. Particularly pretreatment of $Na_2SeO_3$ significantly more compared to the administration of $CH_3HgCl$ only. The concentration of adenosine-5'-triphosphate in $Na_2SeO_3$ treatment groups revealed a favourable effect compared to the administration of $CH_3HgCl$ only. 3. Intracellular $Ca^{2+}$ concentration in administration of $CH_3HgCl$ only was increased significantly more than control group in all test hours but was increased significantly more at 48 hours only after treatment in combined administration of $CH_3HgCl$ and $Na_2SeO_3$ and pretreatment of $Na_2SeO_3$ according to time interval more decreased significantly intracellular $Ca^{2+}$ concentration compared to the administration of $CH_3HgCl$ only. 4. Free intrasynaptosomal $Ca^{2+}$ concentration in the combined administration of $CH_3HgCl$ and $Na_2SeO_3$ was decreased ($24%{\sim}40%$) significantly more than the administration of $CH_3HgCl$ only. From the above results, the specific dosage of $Na_2SeO_3$ decreased increment of intracellular $Ca^{2+}$ concentration induced by administration of $CH_3HgCl$. These findings suggest the protective mechanism of $Na_2SeO_3$ on the neurotoxicity of $CH_3HgCl$.

• Journal of Chest Surgery
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• v.29 no.6
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• pp.606-613
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• 1996

Cardiac transplantation has been the treatment of patients with end-stage heart disease since it was first performed in 1967. In Korea the first case was performed in 1992 and 42 patients underwent heart trans- plantation so far. The purpose of this article is to report short-term result of cardiac transplantation at our center. Between April 1994 and September 1995, 14 patients had undergone orthotopic heart transplantations. There was 12 male and 2 female patients. Mea recipient age was 34 years(range 11 to 54 years) and mean donor age was 28.4 years(16 to 50 years). Mean graft ischemic time was 120.7minutes(80 to 280 minutes). The follow-up period after transplantation was 11 months(3 to 17 months). Recipient diagnosis included dilated cardiomyopathy in 10, ischemic cardiomyopathy in 2, valvular cardiomyopathy in 1, congenital complex heart disease in 1 patient. The preoperative status of the recipients were state I (50%) and ll (50%) by UNOS classification and class 111 (5 patients) and class IV (9) by NYHA functional class. All patients were treated with triple-drug immunosuppression (cyclosporine, azathioprine, steroid) and induction with RATG. The rejection episodes were 5 times in 3 patients during the follow-up. Causes of infection were aspergillosis (2), and hepes zoster (1), CMV pneumonitis (1). Permanent pace- maker was inserted in 1 patient. Currently 9 patients are alive with seven patients in WYHA functional class I and two in class l . The ejection fraction increased from preoperative value of 19.9 $\pm$ 3.4% to postoperative value of 69.0 $\pm$ 5.6%. The causes of death were cellular rejection (1),chronic graft failure due to size-mismatching (1),respirat- oxy insufficiency due to asthma attack (1), subarachnoid hemorrhage (1), and RIO humoral rejection (1).

• Economic and Environmental Geology
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• v.35 no.5
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• pp.395-406
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• 2002

The sixty natural springs and community wells used as a drinking water in the Daejeon area are mainly located at the parks and the natural green districts. The purpose of this study is to investigate the characteristics of water quality and the contamination of the springs and the wells, and to suggest the management strategy for the springs and wells. For this study, we undertook water quality data from Daejeon City. According to the statistic analysis of water quality data, unacceptable rate as a drinking water was about 28 percent in 1999 and 24.5 percent in 2000, respectively. Major unacceptable factor is coliform, and others are bacteria, yersinia, color, turbidity, Fe and F. The unacceptable rate shows a roughly positive relationship with precipitation, that is, it shows highest rate during a rainy season between June and September. The major contamination source is likely to be the excrement of wild animals around natural springs and wells. Most of springs are vulnerable to the contamination of coliform and bacteria because of short residence time and shallow circulation in subsurface environment. The water samples collected from 31 springs or wells show weak acidic pHs, the electrical conductivity ranging from 63 to 357 $\mu\textrm{S}$/cm, and the hydrochemical types of Na(Ca)-HC0$_3$ and Ca-HC0$_3$. The groundwater samples of low total dissolved solid(TDS) belong to Na(Ca)-HC0$_3$. type, and the groundwater of high total dissolved solid is shifted towards Ca-HC0$_3$ type in the chemical composition. These hydrochemical characteristics indicate that most natural springs is in the early stage of geochemical evolution. The natural springs should be closed during a rainy season, which shows a high contamination rate. We suggest that a protection barrier around the springs should be built to keep wild animals away from the springs.

• Proceedings of the Ginseng society Conference
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• 2007.05a
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• pp.23-24
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• 2007

Purpose: 인삼이 항당뇨 활성을 가진다는 연구가 많은 연구자들에 의해 진행되었고, 이는 인삼의 구성 성분 중 ginsenoside에 기인한다는 보고가 있다. 본 연구는 ginsenoside의 항당뇨 작용기전을 in vitro에서 알아보고자 3T3-L1 지방세포에서 glucose uptake와 췌장 베타세포인 HIT-T15 세포에서 insulin 분비 효과를 확인하였다. 이를 위하여 인삼을 식초로 처리한 긴삼의 70% MeOH 분획으로부터 protopanaxadiol 계인 ginsenoside $Rb_2$, $Rg_3$ 그리고 protopanaxtriol 계인 $Rg_2$를 분리하여 본 실험에 사용하였다. Method: Ginsenoside $Rb_2$, $Rg_2$, $Rg_3$가 지방 세포에서 glucose uptake에 미치는 효과를 확인하기 위하여 3T3-L1 세포를 DMEM (Dulbecco's Modified Eagle's Medium) 배지에서 분화 유도시켰으며 3T3-L1 preadipocyte가 80% 정도 자라면 분화 유도 배지 (5% fetal bovine serum (FBS), 0.5 mM isobutylmethylxanthine (IBMX), 1 mM dexamethasone 그리고 $10{\mu}g/ml$ insulin가 포함된 DMEM)로 4일, $10{\mu}g/ml$ insulin가 포함된 DMEM으로 2일, FBS만 포함된 DMEM으로 2일 배양하여 총 8일 동안 분화를 유도하였다. 분화 유도된 3T3-L1 adipocytes 에 각각 $Rb_2$, $Rg_2$, $Rg_3$를 $20{\mu}M$로 처리하여 16시간 배양하여 low glucose DMEM에서 3시간 배양한 후에 $37^{\circ}C$에서 insulin 10 ng/ml 과 각각 $Rb_2$, $Rg_2$, $Rg_3$가 포함된 Krebs Ringer Hepes buffer(KRP buffer)에서 20분간 배양하였다. 2-deoxy-D-[$^3H$]-glucose를 넣고 10분 후에 차가운 PBS로 반응을 종결시켜 lysis buffer로 cell을 모은 후 scintillation counter를 이용하여 glucose를 측정하였다. Insulin 분비 효과는 HIT-T15 세포와 일차 배양한 흰쥐 소도세포(islets)를 사용하여 확인하였다. HIT-T15 세포는 24 well plate에 well 당 $2{\times}10^5$ 개씩 분주하여 24시간 동안 배양한 후 시료를 처리하였으며 소도 세포는 Sprague-Dawley rat의 췌장에 collagenase가 포함된 Hanks' Balanced Salt Solution(HBSS)을 주입하여 분리하고 islets을 얻었다. 분리한 소도세포를 $1{\sim}2$일 동안 배양하여 $Rb_2$, $Rg_2$, $Rg_3$가 각각 $20{\mu}M$의 농도로 첨가된 insulin 측정용 buffer인 Krebs-Ringer buffer (KRB+0.3% BSA, KRBB)에 $37^{\circ}C$에서 1시간 incubation 시킨 후 배양액으로 분비된 인슐린의 양을 측정하였다. 한편 ginsenoside의 인슐린 분비 촉진 기전을 알아보기 위한 실험에서는 ATP-sensitive $K^+$ channel opener인 diazoxide (0.5 mM)가 ginsenoside에 의해 촉진된 인슐린 분비를 억제하는지 살펴보았다. Result: glucose uptake assay 에서는 $Rg_2$가 가장 크게 glucose uptake를 증가시켰고 $Rb_2$, $Rg_3$는 그 활성이 크지 않았다. 한편 Insulin 분비 효과는 diol계인 $Rg_3$에서 용량 의존적으로 인슐린의 분비를 촉진시켰으며 $20{\mu}M$ 농도에서 대조군과 비교해 1.5배 이상의 분비 촉진 효과를 보였고 triol계인 $Rg_2$ 에서는 이러한 효과가 나타나지 않았다. $Rg_3$의 인슐린 분비 촉진 기전을 0.5 mM 의 diazoxide를 이용하여 확인한 결과 $Rg_3$에 의해 촉진된 인슐린 분비를 감소시켰다. 이로 미루어보아 $Rg_3$의 인슐린 분비 촉진 기전은 ATP-sensitive $K^+$ 채널의 봉쇄에 의한 것임을 확인할 수 있었다.