• Journal of Embryo Transfer
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• v.31 no.3
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• pp.169-177
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• 2016

There is a growing interest in the application of primary hepatocytes for treatment of liver diseases in humans and for drug development. Several studies have focused on long-term survival and di-differentiation blocking of primary hepatocytes in an in vitro culture system. Therefore, the present study also aimed to optimize an in vitro culture system using primary rat hepatocytes. Primary rat hepatocytes from 6-week-old male Crl:CD rats were isolated using a modified two-step collagenase perfusion. Healthy $3.5{\times}10^6$ primary rat hepatocytes were seeded into a 2 dimensional (2D) culture in a 25T culture flask coated with collagen type I or into a 3D culture in a 125-ml spinner flask for 7 days. Production of plasma protein (ALB and TF), apoptosis (BAX and BCL2), and CYP (CYP3A1) related genes were compared between the 2D and 3D culture systems. The 3D culture system had an advantage over the 2D system because of the relatively high expression of ALB and low expression of BAX in the 3D system. However, the level of CYP3A1 did not improve in the 3D culture with and without the presence of a dexamethasone inducer. Therefore, 3D culture has an advantage for albumin production and primary rat hepatocyte survivability, but a low expression of CYP3A1 indicated that primary rat hepatocytes require a high-density culture for stress reduction by continuous flow.

• The Korean Journal of Physiology
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• v.4 no.2
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• pp.33-36
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• 1970

A study was planned to see if the ginseng has any influence upon the weight of viscera of male and female rats. One hundred and ten male rats (body weight: $35{\sim}40\;gm$ at the beginning of the experiment) and an equal number of females with similar body weight were use4 Excluding 10 male and 10 females which served as controls, these rats were divided equally into 4 groups (male ginseng group, male saline group, female ginseng group and female saline group), each of which comprising 50 animals. For 45 days, the male and female ginseng groups received every day 0.6 m1/100 gm body weight of ginseng extract solution (4 mg of ginseng alcohol extract in 1 ml of saline) subcutaneously, and the male and female saline groups received the same amount of physiological saline. Each group was further divided into 5 subgroups comprising 10 animals, and the 10 control rats and each subgroups were sacrificed immediately before, and 5, 15, 35, 35, and 45 days after beginning of the drug administration, respectively. The body weight of each subgroup was checked immediately before sacrificing it, and the heart, lung, liver, kidney, adrenal gland, spleen, and testis or uterus of each animal were weighed with a torsion balance. Following results were obtained: 1. Ginseng significantly facilitated gain in weight of testis and spleen as well as body weight of rats from 35th day after beginning of the drug administration on, but not before this time. 2. Ginseng did not exert significant influence upon the weight of heart, lung, liver, tidney, adrenal gland, and uterus of the rat.

• The Journal of Internal Korean Medicine
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• v.18 no.2
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• pp.1-26
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• 1997

Chunggeumtang has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Chunggeumtang on tracheal smooth muscle is not konwn. The purpose of the present study is to determine the effect of Chunggeumtang on histamine and acetylcholine induced tracheal smooth muscle contraction in rats and guinea pigs. Guinea pig(500g, male) and Sprague Dawley rats (250g, male) were killed by $CO_2$ exposure and a segment (8-10mm) of the thoracic trachea from each rat and guinea pig was cut into equal swegments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force diplacement transducers under 0.5g loading tension. The dose of histamine (His) and acetylcholine (Ach) which evoked 50% of maximal response ($ED_{50}$) was obtained from cumulative dose response curves for histamine and acetylcholine($10^{-7}{\sim}10^{-4}M$). Contractions evoked by His ($ED_{50}$) and Ach ($ED_{50}$) were inhibited significantly by Chunggeumtang. In guinea pig tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $6.1%after\;30{\mu}l/ml$ Chunggeumtang, and 49.4% (p<0.01) after $100{\mu}l/ml$ Chunggeumtang. In rat tracheal smooth muscle, the mean percent inhibition of acetylcholine induced contraction was $6.7%\;after\;30{\mu}l/ml$ Chunggeumtang, and $54.2%\;(p<0.01)\;after\;100{\mu}l/ml$ Chunggeumtang. Also, in guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $30.6%\;(p<0.05)\;after\;30{\mu}l/ml$ Chunggeumtang, and $53.0%\;(p<0.01)\;after\;100{\mu}l/ml$ Chunggeumtang. In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was $24.1%\;(p<0.05)\;after\;30{\mu}l/ml$ Chunggeumtang, and $55.3%\;(p<0.01)\;after\;100{\mu}l/ml$ Chunggeumtang. Propranolol and indomethacin($10^{-7}M$) slightly but significantly attenuated the inhibitory effects of Chunggeumtang. Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$ Chunggeumtang fell to 27.6% in guinea pig induced by acetylcholine contraction and by $100{\mu}l/ml$ Chunggeumtang fell to 28.7% (p<0.05) in rat induced by acetylcholine contraction and the mean percent inhibition caused by $100{\mu}l/ml$ Chunggeumtang fell to 16.2% in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Chunggeumtang fell to 28.7% (p<0.05) in rat induced by histamine contraction. Indomethacin, the mean percent inhibition caused by $100{\mu}l/ml$ Chunggeumtang fell to 20.0% in guinea pig induced by acetylcholine contraction and by $100{\mu}l/ml$ Chunggeumtang fell to 16.9% (p<0.05) in rat induced by acetylcholine contraction and the mean percent inhibition caused by $100{\mu}l/ml$ Chunggeumtang fell to 16.4% in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Chunggeumtang fell to 23.1% (p<0.05) in rat induced by histamine contraction. Methylene blue($10^{-7}M$) did not significantly alter the inhibitory effect of Chunggeumtang. Also, I could find the effects of Chunggeumtang and Chunggeumtanggamorphine on the tracheal smooth muscle in guinea pig and rat did not change significantly. These results indicate that Chunggeumtang can relax histamine and acetylcholine-induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects and the release of cyclooxygenase products.

• Journal of Radiation Protection and Research
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• v.32 no.4
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• pp.184-189
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• 2007

In order to investigate the enhancement effects of low dose radiation on biological activation, this study applied low dose X-ray to the whole body of male rats to find out whether hormesis is induced in male germ cells. Total 36 Sprague-Dawley(SD) rats as experimental animal were subdivided into 6 groups(in 6 rats per group) such as control, 10 mGy, 20 mGy, 50 mGy, 100 mGy and 200 mGy radiation group All the groups showed slightly increasing number of sperms per 0.1g semen ($14.216{\times}10^6,\;13.901{\times}10^6,\;14.153{\times}10^6,\;13.831{\times}10^6,\;14.137{\times}10^6,\;14.677{\times}10^6$ respectively), and the motility of sperms amounted to 50.9%, 49.5%, 55.1%, 54.3%, 48.0% and 52.2% respectively. Particularly, compared to the control, the other 5 groups showed higher male hormone level, and the microscopic observations of testicle tissues showed no vacuolization in seminiferous tubules and testis cells. In the results of this experiment, no harmful effect was observed on Sprague-Dawley (SD) rats for which the dose of radiation was controlled as regulated legally by the Ministry of Science and Technology and the Ministry of Health and Welfare. However, as these results were obtained from a limited number of animals, we cannot maintain that the same effect will be observed in the human body. Therefore, there should be further research on the effect on other animals and ultimately on the human body.

• Toxicological Research
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• v.29 no.2
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• pp.91-98
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• 2013

Armeniacae semen (AS) has been considered a toxic herb in the Korean medicine as it contains hydrogen cyanide and amygdalin, especially in its endocarp. Therefore, prebrewed AS that is devoid of endocarp has been traditionally used. In the present study, amygdalin content of the prebrewed AS was significantly lower ($2.73{\pm}0.32{\mu}g/ml$; p<0.01) than the content in the extract that contained the endocarps ($28.50{\pm}6.71{\mu}g/ml$); amygdalin content corresponded to 10% of the extract in the present study. Because of single oral dose toxicity of prebrewed AS according to the recommendation of Korea Food and Drug Administration Guidelines (2009-116, 2009), which was based on single oral dose toxicity study of prebrewed AS, mortality due to toxic principles was significantly reduced. In this study, 2,000 mg/kg of prebrewed AS led to death of 1 female rat and 1 male rat at the end of 2 hr of administration. Based on these results, the 50% lethal dose in both male and female rats was determined to be 9279.5 mg/kg. Seizure, loss of locomotion, and increases in respiration and heart rate were observed as prebrewed AS treatment-related toxicological signs; these signs were restrictedly manifested in the prebrewed AS (2,000 mg/kg)-treated rats. In addition, no changes were observed in body weight, organ weight, gross features, and histopathological parameters with 2,000 mg/kg of AS in both male and female rats. These findings serve as direct evidence that amygdalin in AS is the toxic principle, which can be reduced by the traditional prebrewing method involving the exclusion of endocarp.

• The Korea Journal of Herbology
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• v.37 no.3
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• pp.21-27
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• 2022

Objectives : Single oral dose toxicity test of Lythri Herba water extracts (LHWE) in Sprague-Dawley (SD) rat was performed to determine approximate lethal dose (ALD) of LHWE. Methods : This test was progressed according to OECD Guidelines for the Testing of Chemicals : acute oral toxicity. After adaptation of 7 days, SD rats were divided into 2 groups : vehicle control and 5000 mg/kg LHWE-treated group. Each group consisted of 5 female rats and 5 male rats. Vehicle or 5000 mg/kg LHWE was orally administrated once a day. Survival rates, general toxicity, and changes of body weight were investigated for 14 days after administration. On the last day of examination, the weight of all animals was measured and an autopsy was performed. All internal organ abnormalities were checked macroscopically and their findings were recorded. Results : In both groups, dead animals were not observed. During 14 days of administration, abnormal clinical signs were not detected. There was also no significant difference in weight gains between each group. Autopsy analysis showed that one case of the LHWE-treated female group had retention of clear fluid in the uterus; however, it was not considered to be affected by LHWE administration. Moreover, abnormal findings were not discovered in the control male group and the LHWE-treated male group. Conclusions : These results suggest that the ALD of LHWE exceed 5000 mg/kg and single oral administration of LHWE below 5000 mg/kg is nontoxic.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.16 no.3
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• pp.233-244
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• 2006

Of many volatile organic detergents for metals, benzine(CAS No. 8030-30-6), of which the toxicity has not yet been proven, has been used as an alternative of the halide compounds in the consideration of toxic effects, global warming and the destruction of ozone layer. In order to evaluate the effects of the benzine on human body by investigating the subchronic inhalation toxicity, to obtain the basic data for establishing the criteria of exposure in working environments and to classify the hazardousness in compliance with the Industrial Safety and Health Act by evaluating the hazardousness, repeated inhalation exposure test was carried with SD rats. The rats were grouped by 10 females and males each. The repetitive inhalation exposures were carried out at 4 levels of concentration of 0 ppm, 60 ppm, 300 ppm, and 1,500 ppm, for 6 hours a day, 5 days a week, for 13 weeks. The results are described hereunder. 1. No death of the animals of the exposed and controlled groups in the test period. Not any specific clinical symptoms, change in feed intake quantity, abnormality in eye test, or change in activity were observed. 2. In the 300 ppm and 1,500 ppm groups, weight reduction in the female groups and weight increase of liver and kidney in the male groups compared with control group were observed with statistical significance(p<0.05). 3. In the blood test, the HCT increased in the male 300 ppm group and the number of hematocyte increased, MCV and MCH decreased in the male 1,500 ppm group. In the female 1,500 ppm group, the HB decreased and the distribution width of the hematocyte particle size increased. In the blood biochemistry test, the TP in the male 1,500 ppm group and the LDH in the female 1,500 ppm group were increased with statistical significance(p<0.05). 4. Under the test conditions of the present study with SD rats, the NOEL was evaluated to be from 60 ppm to 300 ppm for both male and female groups. By extrapolation, the NOEL for human who work 8 hours a day was evaluated to be from 128 ppm to 640 ppm 5. Since the NOEL evaluated in this study do not exceed 60ppm(0.184 mg/L) the test material does not belong to the classification of the hazardous substance "NOEL${\leq}$0.5mg/L/6hr/90day(rat), for continuous inhalation of 6hours a day for 90 days" nor to the basic hazardous chemical substance class 1(0.2 mg/L/6hr/90day(rat) defined by the GHS which is a criteria of classification and identification of chemical compounds. However, considering the boiling point($30-204^{\circ}C$), flashing point($-40^{\circ}C$), vapor pressure(40 mmHg), and the inflammable range(1.0 - 6.0 %), sufficient care should be taken for handling in the safety aspects including fire or explosion.

• Development and Reproduction
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• v.4 no.1
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• pp.95-100
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• 2000

A large number of man-made chemicals that have been released into the environment have the potential to disrupt the endocrine system of animals and humans. There is a critical developmental period during which sexual brain differentiation proceeds irreversibly under the influence of gonadal hormone. Recently we identified KAP3 gene expressed during the critical period of rat brain sexual differentiation. KAP3 functions as a microtubule-based motor that transports membranous organelles anterogradely in cells, including neurons. In the present study, we aimed to investigate the effect of endocrine disrupter, Dichlorodiphenyl trichloroethane (DDT), on the KAP3 gene expression during critical period of rat brain development. Maternal exposure to DDT increased the level of KAP3 mRNA in male and female fetus brains when examined on the gestational day 17 (GDl7). In postnatal day 6, DDT suppressed the expression of KAP3 gene in male and female rat brain. Also, the body weight and fertilization rate were decreased in the DDT exposured rats. These results showed that endocrine disrupter, DDT, can affect the transcriptional level of brain sexual differentiation related gene, KAP3, in the prenatal and the neonatal rat brain and that maternal exposure to endocrine disruptors may lead to a toxic response in embryonic differentiation of brain. And so KAP3 gene may be used a gene maker to analyse the molecular mechanism for toxic response in animal nerve tissues exposed to endocrine disruptors.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.57-69
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• 2003

It is well known that many pesticides possess hormonal activity, and affect the developments of wildlife and mammals including human. Currently, pyrethroid insecticides are in worldwide use to control in and outdoor pests, providing potential far environmental exposure. Hormonal activities of these pyrethroid insecticides, however, have been little studied, and the developmental effects of them were no reported. Therefore, we firstly examined the potential estrogenic activities of some pyrethroid insecticides (permethrin, cypermethrin, tetramethrin, deltamethrin, sumithrin, fenvalerate and bioallethrin) by immature rat uterotrophic assay, luciferase reporter gene assay and Calbindin-D$\sub$9k/ (CaBP-9k) gene expression assay. Uterine wet weights were increased by permethrin and the permethrin-induced weights were inhibited by ICI 182780 in the uterolrophic assay. On the other hand tetramethrin significantly reduced uterine and vaginal wet weights, and also inhibited the E2-induced weight increases at all doses tested. Cypermethrin and sumithrin had a tendency to increase uterine weights, although not statistically significant. Permethrin and cypermethrin dose-dependently increased the luciferase activity in reporter gene assay. Northern blot analysis showed that permethrin induced CaBP-9k mRNA expression whereas tetramethrin inhibted. Subsequent studies were conducted to investigate the possible developmental effects of four pyrethroid insecricides (permethrin, cypermethrin, sumithrin and teramethrin). Either diethlbestrol (DES) or 17${\beta}$ -estradiol (E2) was used as a reference control in this study. Pyrethroid insecticides were administered to Sprague Dawley rats via subcutaneous injection at 6 to 18 days of gestation or 1 to 5 days after birth. In utero treatment of permethrin (10mg/kg/day) in female rat resulted in significant increases in uterine and ovarian weights while significant decreases in serum E2 concentration, uterine and ovarian ER${\alpha}$ mRNA levels. Sumithrin and permethrin led to acceleration in vaginal opening of female rat, while delay in preputial separation of male after neonatal treatment. Anogenital distances of PND 18 were significantly reduced in sumthrin-treated, and permerhrin-treated male rats after neonatal treatment. All the pyrethroid insecticides tested caused significant increases in uterine weights on PND 18, while significant reductions in the first diestrus phase when neonataly treated. In addition, exposure to pyrethroids in neonatal period led to significant reduction in relative brain weight in female rat on PND 18, but its weight was recovered in diestrus phase. In summary, Our experimental data demonstrate the possibilities of developmental effects of pyrethroid insecticides via estrogenic or antiestrogenic activity.

• Korean Journal of Acupuncture
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• v.18 no.1
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• pp.157-164
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• 2001

Radix Asteris has been used in Korea for many centuries as a treatment for respiratory disease. The effect of Radix Asteris on tracheal smooth muscle is not known. The purpose of the present study is to determine the effect of Radix Asteris on histamine induced tracheal smooth muscle contraction in rats and guinea pigs. Guinea pig(500g, male) and Sprague Dawley rats (250g, male) were killed by $CO_2$ exposure and a segment (8-10mm) of the thoracic trachea from each rat and guinea pig was cut into equal segments and mounted 'in pairs' in a tissue bath. Contractile force was measured with force displacement transducers under 0.5g loading tension. The dose of histamine (His) which evoked 50% of maximal response ($ED_{50}$) was obtained from cumulative dose response curves for histamine $(10^{-7}{\sim}10^{-4}M)$. Contractions evoked by His ($ED_{50}$) were inhibited significantly by Radix Asteris. In guinea pig tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was 120.5% (p<0.01) after $100{\mu}l/ml$ Radix Asteris. In rat tracheal smooth muscle, the mean percent inhibition of histamine induced contraction was 135.4% (p<0.01) after $100{\mu}l/ml$ Radix Asteris. Propranolol $(10^{-7}M)$ slightly but significantly attenuated the inhibitory effects of Radix Asteris. Following treatment with propranolol, the mean percent inhibition caused by $100{\mu}l/ml$ Radix Asteris fell to 44.6% in guinea pig induced by histamine contraction and by $100{\mu}l/ml$ Radix Asteris fell to 18.7% (p<0.05) in rat induced by histamine contraction. Indomethacin and methylene blue$(10^{-7}M)$ did not significantly alter the inhibitory effect of Radix Asteris. These results indicate that Radix Asteris can relax histamine induced contraction of guinea pig and rat tracheal smooth muscle, and that this inhibition involves sympathetic effects.