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Antrum Formation and Growth of Mouse Pre-antral Follicles Cultured in Two Different Culture Media without Hormones

  • Kim, Ju-Hwan;Kim, Hwan-Tae;Park, Kee-Sang;Song, Hai-Bum;Chun, Sang-Sik
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.8-8
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    • 2001
  • Mouse follicles require the addition of gonadotropins (Gns) to complete maturation and ovulation of oocyte and antrum formation of follicles in vitro. However, we tried examination of in vitro growth of mouse pre-antral follicles in medium without Gns and physiological factors. And also, pre-antral follicles were isolated from ovaries by mechanical method. Our present studies were conducted to evaluate on the growth of follicles and intra-follicular oocytes and antrum formation in vitro of mouse pre-antral follicles in two different media. Pre-antral follicles (91-120${\mu}{\textrm}{m}$) were isolated mechanically by fine 30G needles not using enzymes from ovary of 3-6 weeks old female ICR mice. Isolated pre-antral follicles were cultured in 20 ${mu}ell$ droplets of TCM (n=17; follicles: 107.8 $\pm$ 1.58 ${\mu}{\textrm}{m}$; oocytes: 59.9$\pm$1.2 ${\mu}{\textrm}{m}$) or MEM (n=12; follicles: 109.3$\pm$2.53 ${\mu}{\textrm}{m}$; oocytes: 55.4 $\pm$1.6${\mu}{\textrm}{m}$) under mineral oil on the 60mm culture dish. All experimental media was supplemented with 10% FBS but without Gns and/or physiological factors. Pre-antral follicles were individually cultured in drops for 8 days. Antrum formation and growth of pre-antral follicles and intra-follicular oocytes were evaluated using a precalibrated ocular micrometer at $\times$200 magnifications during in vitro culture. Results between different groups were analyzed using combination of Student's t-test and Chi-square, and considered statistically significant when P<0.05. Antrum formation of pre-antral follicles had started in two culture media on day-2. On day-8, antrum formation had occurred in 58.3%(7/12) of pre-antral follicles cultured in MEM, but only in 23.5% (4/17) of those cultured in TCM (P=0.0364). Growth of pre-antral follicles and intra-follicular oocytes were observed on day-4 and -8. On day-4, follicular diameters was similar (P=0.1338) in TCM (119.4$\pm$2.58 ${\mu}{\textrm}{m}$) and MEM (125.4$\pm$4.52 ${\mu}{\textrm}{m}$). However, on day-8, diameters of pre-antral follicles cultured in MEM (168.9$\pm$17.29 ${\mu}{\textrm}{m}$) was significantly (P=0.0248) bigger than that in TCM (126.7$\pm$4.28 ${\mu}{\textrm}{m}$). On day-4 and -8, diameters of intra-follicular oocytes were similar TCM (67.1$\pm$1.3 and 72.4$\pm$0.9${\mu}{\textrm}{m}$) and MEM (65.2$\pm$1.7 and 73.3$\pm$1.5 ${\mu}{\textrm}{m}$), respectively. We can conform that medium not supplemented with Gns and/or physiological factors can be used for in vitro antrum formation and growth of mouse pre-antral follicles and intra-follicular oocytes. In conclusion, MEM supplemented with FBS can be used for growth in vitro of mouse pre-antral follicles isolated mechanically.

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Growth enhancement and cytotoxicity of Korean mistletoe fractions on human cell lines (한국산 겨우살이 분획물의 면역세포의 생육증진 및 세포독성)

  • Lee, So-Jin;Lee, Mi-Kyoung;Choi, Geun-Pyo;Yu, Chang-Yeon;Roh, Seong-Kyu;Kim, Jong-Dai;Lee, Hyeon-Yong;Lee, Jin-Ha
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.1
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    • pp.62-70
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    • 2003
  • The biological activities on human immune and cancer cell lines of the four kinds of Korean mistletoes (Korean Viscum album, var. coloratum, : Korean Viscum sp. in Quercus acutissima Carr., Korean Viscum sp. in Castanea crenata, Korean Viscum sp. in Betula platyphylla, and Korean Viscum sp. in Salix koreensis) extracts were investigated. The extracts were preparated with ethanol, and fractionated with n-butanol, ethyl acetate, chloroform, hexane, and second distilled water. Cytotoxic potencies of the fractions on human normal lung cell line (HEL 299) showed under 28% in the concentration of 0.5 mg/ml. Growth inhibition effect of the Korean mistletoe extracts on the several human cancer cell lines depends on the concentration of the extracts, and extracting solvent. The hexane, chloroform, and ethyl acetate fractions indicated a strong anticancer activity, but not in aqueous and butanol fractions. Some mistletoe fractions have a different characteristic on the cancer cell lines. Stimulation on the growth of human immuno cell lines(B cell : Raji, T cell: Jurkat) of the extracts were confirmed in the ethyl acetate, chloroform, hexane fractions, but not in aqueous system.

Health Assessment of Shift Workers in a Automobile Manufacturing Plant (자동차공장 교대작업 근로자들의 건강상태평가)

  • Lee, Jung-Jeung;Chung, Jong-Hak
    • Journal of Preventive Medicine and Public Health
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    • v.28 no.1 s.49
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    • pp.103-121
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    • 1995
  • An investigation on psychophysiological health and social well-being of shift workers been carried out on workers of a automobile manufacturing plant in Ulsan, for 1 month from April, 1993. This cross-sectional survey compared shift workers(n=544) with day workers(n=115). Each subject completed a questionnaire about his personal habit, background, shift schedule, sleep and eating patterns, subjective digestive symptom and psychological well-being and distress using the General Well-Being Schedule(GWB) by self administrated questionnaire that was developed for the U.S. Health and Nutrition Examination Surveys (HANES I). Chi-square analysis was used for discontinuous data and the t-test was used for continous data to determine whether differences noted between the two groups. In terms of sleep quality, a greater percentage of shift workers frequently had trouble getting back to sleep once awakend(p<0.01), and a greater percentage of shift workers awakend tired or sleepy frequently more often than day workers(p<0.05). In rating the quality of their sleep, more shift workers rated their sleep fair to poor than day workers, and greater percentage of shift workers felt tired or sleepy at work two or more times per week(p<0.01) and a much higher percentage of shift workers felt tired or sleepy after work every days(p<0.01). In terms of sleep patterns, a much higher percentage of day workers reported uninterrupted sleep per 24hours than shift workers. The shift workers reported different eating patterns from day workers but there were no statistically significant and rate of their appetite. Thirteen percent of day workers reported the best appetite but only 6.6% of shift workers had the best appetite. The gastric complaints is more frequent in shift workers than day workers(p<0.01). Among subscores in General Well-Being Schedule, anxiety, depression, positive well-being and vitality subscale of shift workers were lower than those of day workers(p<0.05) and general health and self control subscale of shift workers were lower than .those of day workers but there were no statistical significant difference. Based on these study result, it could be concluded that the shift work has significant effects on some psychophysiological conditions of the workers.

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A Study on Knowledge, Attitude and Behavior to Sexuality in Middle School Students (중학생의 성에 대한 지식, 태도, 행위에 관한 조사연구)

  • Cha, Eun-Seok
    • Journal of the Korean Society of School Health
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    • v.12 no.2
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    • pp.357-375
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    • 1999
  • The purpose of this study is to find out methods to control, regulate and prevent sexual violence and provide guidelines for sex education in middle school. This study was conducted to examine factors in relation to sexuality, attitudes toward women, experience of sexual contact, frequencies and kinds of pornography with which they had contact, and sex education. For the data collection, a survey was conducted from September 25 to October 1, 1997 by using prepared questionnaires. Questionnaires were given to 503 students from three different kinds of middle schools in Seoul, Boys, Girls, and Coeducation schools. The data analyses used a PC-SAS. Each item was examined by frequencies and percentage. To compare Boys with Girls, I applied a T-test. In addition, with the kinds of school type, I applied anANOVA, ${\chi}^2$. Moreover, For investigating and analyzing each category, I categorized knowledge and attitude of sexuality, with precautionary measures of sexual violence. After that I applied T-test, ANOVA, and a Pearson's correlation for each category. The instruments of this study consisted of 9 domains. (1) General characteristics of the respondents (2) 9 questions about the interaction between he/she and his/her parent. (3) 12 questions, concerning A Scale for Attitudes of Adolescents Toward women(AWSA). (4) 26 questions related to sexual knowledge. (5) 25 questions about sexual attitude (6) strategies for prevention of sexual violence-33 questions (7) 5 questions related to sexual behavior (8) the experience of contact with pronography. (9) the experience of sex-education. The major results were summarized as follows; 1. Like many researches, This study shows that the girls are more eqalitarian than boys. The girls mean ($40.90{\pm}3.67$) is significantly higher than boys ($34.72{\pm}3.77$). Most Koreans have believed that there are many differences between men's roles and women's ones, because of confucianism which had been implied in most thought and ideas since the Lee dynasty. Therefore, the result is not surprising at all. 2. Most students answered correctly the questions of sexual knowledge. The girl's score was higher ($Mean{\pm}SD$ : $48.88{\pm}2.39$) than the boys' ($Mean{\pm}SD$ : $46.95{\pm}3.90$)(p 0.001). Nevertheless, the hypothesis that sexual knowledge influences sexual attitude and its behavior was rejected. 3. When twenty-five questions were asked about sexual attitude, the girl students investigated were found to have a more propre sexual attitude than that of the boys. A significant difference was found between boys and girls(p=0.001). The result shows that a person who has more experience in viewing pornography and who smokes will have a more distorted view of sexuality. 4. There are many students who have experienced of sexual contact. They need to take a sex education program about contraception and prevention of sexually transmitted diseases. Systematic and concrete sex education is one way to lead them to proper sexual behavior. 5. Most respondents rejected stereotyped attitudes towards women. Boys, however, were more likely to accept stereotypes of Women. The girl's mean($Mean{\pm}SD$ : $40.90{\pm}3.67$) is higher than the boys($Mean{\pm}SD$ : $34.72{\pm}3.77$)(p=0.0001). 6. Many boys ignored the rules for prevention of sexual violence. A boy or a girl who has more experience of sexual contact over kiss and sex-typed neglected the rules, too(p=0.001). Today, there is increasing juvenile delinquency related to sexuality. This study provided the basic material for a preventive education system. I believe that a systematic and concrete sex education system can be helpful for adolescents and promote their responsibility as well as cultivate morality about sexuality. As a result, juvenile delinquency can be decreased. This study is basically aimed to provide information for a prevenative education system of sexual violence. Further research is recommended to evaluate programs in schools.

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Yield of Green Manure and Nitrogen of Cornflower (Centaurea cyanus L.) in Different Upland Soil Textures (토성별 수레국화 (Centaurea cyanus L.)의 녹비수량 및 질소생산량)

  • Cho, Hyeoun-Suk;Seong, Ki-Yeung;Park, Tae-Seon;Seo, Myung-Chul;Jeon, Weon-Tai;Kang, Hang-Won;Lee, Hye-Jin
    • Korean Journal of Soil Science and Fertilizer
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    • v.45 no.4
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    • pp.664-670
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    • 2012
  • Experiments were conducted to find out the landscape effects and green manure production at the same time in farmland. Cornflower was grown in different soil texture with sand, sandy loam, loam, clay loam, and was sowing with autumn and spring respectively. The overwintering rate of cornflower was at 58.7% in average, and the treatment at sand soil showed 62.1% that was highest among other soils, which cornflower is possible to winter landscape crop. After flowering of cornflower, the contents of total nitrogen (T-N) and total carbon (T-C) in plant were 15.0 and $409.2g\;kg^{-1}$, respectively, and the carbon-nitrogen ratio (C/N) was 28.6. The yield of cornflower biomass, which will be returned to soil as green manure, recorded $1,210{\sim}3,920kg\;ha^{-1}$ at the spring seeding higher than the autumn seeding as $1,540{\sim}3,170kg\;ha^{-1}$, and the biomass treated by soil texture were showed that the treatments at the clay loam had been the largest yields both spring and autumn seeding among at other treatment of soil. The heights of cornflower regardless of soil treatments were 52.8 to 73.6 cm at the autumn seeding and 35.5 to 79.2 cm at the spring seeding although it was more significant variation at the soil textures than the seeding periods. The flowering periods of cornflower ranged from $17^{th}$ to $20^{th}$ in May at the autumn seeding and from $19^{th}$ to $20^{th}$ in June at the spring seeding, which was faster 30 days approximately at the autumn seeding than the spring seeding. In a view of the cornflower application as green manure after flowering, the autumn seeding, when considered to combine with following crops, was more suitable and various than the spring seeding, even though the yield at spring seeding was higher than one at autumn seeding.

Antioxidant Effects of Cysteine-containing Peptides of Different Lengths in Human HaCaT Keratinocytes Exposed to Hydrogen Peroxide (과산화수소에 노출된 인간 각질형성세포에서 길이가 다른 시스테인 함유 펩타이드의 항산화 효과)

  • Jae Won Ha;Joon Yong Choi;Yong Chool Boo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.49 no.3
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    • pp.193-201
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    • 2023
  • Hydrogen peroxide (H2O2) is a type of active oxygen species (ROS) that causes oxidative stress in cells and affects cell growth, proliferation, senescence, and death. The purpose of this study is to find active peptides that attenuate cytotoxicity of H2O2. A positional scanning synthetic tetrapeptide combinatorial library was screened to predict the sequence of potentially active peptides. As a result of comparing the effect of peptide pools on H2O2-induced death of human keratinocytes (HaCaT cells), various active peptide sequences were predicted. Especially, peptides containing cysteine (C) residue were predicted to be active. In follow-up experiments, the cytotoxicity and activity of cysteine-containing peptides of different lengths, such as C-NH2, CC-NH2, CCC-NH2, and CCCC-NH2 were examined. C-NH2 and CC-NH2 showed no significant cytotoxicity up to 1.0 mM, but CCC-NH2, and CCCC-NH2 showed relatively strong cytotoxicity. C-NH2 and CC-NH2 alleviated H2O2-induced cytotoxicity. CC-NH2 was more cytoprotective compared to C-NH2, C, N-acetyl cysteine (NAC), and glutathione (GSH). When intracellular ROS was measured by flow cytometry, H2O2 increased ROS production, and CC-NH2 suppressed ROS production more effectively than C-NH2, and it was as effective as C, NAC, and GSH. This study suggests that CC-NH2 of the cysteine-containing peptides of different lengths has an antioxidant property that safely and effectively alleviates H2O2-induced cytotoxicity and ROS production.

Effect of Fineness of Siliceous Materials on Correction of Soil Acidity under Submerged Condition (담수시(湛水時) 규산물질(珪酸物質)들의 입도별(粒度別) 토양산도교정능력(土壤酸度矯正能力))

  • Lee, Yun Hwan;Han, Ki Hak;Kim, Bok Jin
    • Korean Journal of Soil Science and Fertilizer
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    • v.5 no.2
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    • pp.59-64
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    • 1972
  • The liming effect of the four different siliceous materials with six grades of fineness were investigated in comparison with limestone under the submerged condition for three months, and the alkalinity extracted by 0.07N-EDTA and N-NaOAc solutions were determined to evaluate the neutralizing power of these materials. 1. Fused phosphate took 20 days with finer particles than 60 mesh (Tyler), 34 days with -40+48 mesh particles and 84 days with -20+25 mesh particles to reach the pH 5.5 from pH 4.0 of initial soil pH. These adjusted soil acidities were less 1.0-0.5 unit of pH compared with the pH of particles of limestone. 2. The basic reduction furnace slag increased the pH value to 5.5 in the finer particles than 100 mesh, but the other coarse particles appeared to have slow changes of the soil acidity to pH 4.5-4.7 for the three months. Wollastonite didn't affect the increase of soil pH in coarser particles than 25 mesh whereas other finer particles increased upto pH 4.5-5.0. Blast furnace slag is definitely slower through all sizes of particles. 3. In the relationship between the adjusted soil acidities and alkalinities dissolved in EDTA and NaOAc solution, NaOAc-alkalinity agreed quite closely to the activity of neutralizing value of silicates and limestone containing fineness of particles. The correlation coefficients between the amended soil acidities and NaOAc-alkalinities were stabilized with high significance at the 8 days after water logging and 16 days with the EDTA-alkalinities.

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Comparable Ablation Efficiency of 30 and 100 mCi of I-131 for Low to Intermediate Risk Thyroid Cancers Using Triple Negative Criteria

  • Fatima, Nosheen;Zaman, Maseeh uz;Zaman, Areeba;Zaman, Unaiza;Tahseen, Rabia
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1115-1118
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    • 2016
  • Background: There is controversy about ablation efficacy of low or high doses of radioiodine-131 (RAI) in patients with differentiated thyroid cancers (DTC). The purpose of this prospective study was to determine efficacy of 30 mCi and 100 mCi of RAI to achieve successful ablation in patients with low to intermediate risk DTC. Materials and Methods: This prospective cross sectional study was conducted from April 2013 to November 2015. Inclusion criteria were patients of either gender, 18 years or older, having low to intermediate risk papillary and follicular thyroid cancers with T1-3, N0/N1/Nx but no evidence of distant metastasis. Thirty-nine patients were administered 30 mCi of RAI while 61 patients were given 100 mCi. Informed consent was acquired from all patients and counseling was done by nuclear physicians regarding benefits and possible side effects of RAI. After an average of 6 months (range 6-16 months; 2-3 weeks after thyroxin withdrawal), these patients were followed up for stimulated TSH, thyroglobulin (sTg) and thyroglobulin antibodies, ultrasound neck (U/S) and a diagnostic whole body iodine scan (WBIS) for ablation outcome. Successful ablation was concluded with stimulated Tg< 2ng/ml with negative antibodies, negative U/S and a negative diagnostic WBIS (triple negative criteria). ROC curve analysis was used to find diagnostic strength of baseline sTg to predict successful ablation. Results: Successful ablation based upon triple negative criteria was 56% in the low dose and 57% in the high dose group (non-significant difference). Based on a single criterion (follow-up sTg<2 ng/ml), values were 82% and 77% (again non-significant). The ROC curve revealed that a baseline sTg level ${\leq}7.4ng/ml$ had the highest diagnostic strength to predict successful ablation in all patients. Conclusions: We conclude that 30 mCi of RAI has similar ablation success to 100 mCi dose in patients with low to intermediate risk DTC. A baseline $sTg{\leq}7.4ng/ml$ is a strong predictor of successful ablation in all patients. Low dose RAI is safer, more cost effective and more convenient for patients and healthcare providers.

Comparison of immune cell populations in bronchoalveolar lavage cells and PBMC cytokine expressions in porcine reproductive and respiratory syndrome and porcine respiratory disease complex

  • Yang, Myeon-Sik;Jeong, Chang-Gi;Nazki, Salik;Mattoo, Sameer ul Salam;Lee, Sang-Myeong;Kim, Won-Il;Kim, Bumseok
    • Korean Journal of Veterinary Service
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    • v.42 no.4
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    • pp.201-216
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    • 2019
  • Porcine reproductive and respiratory syndrome (PRRS) is characterized by reproductive failure in sows and respiratory distress in all age pigs. Porcine respiratory disease complex (PRDC) is a disease caused by opportunistic bacterial infection secondary to a weakened immune system by a preceding respiratory infection. In this study, we tried to compare the immune responses in PRRS and PRDC groups to clearly characterize the disease severity. Eighty-five pigs were infected with various Korean field PRRS virus strains. Infected animals were classified into PRRS (n=32) and PRDC (n=53) groups based on lung lesions such as interstitial pneumonia, suppurative pneumonia, and pleuropneumonia. The immune cell population of bronchoalveolar lavage cells (BALc) was evaluated on 14 and 28 days post infection (dpi) and PMBC cytokine expression was measured on 0, 3, 7, 14 dpi to investigate early inflammatory reactions. Pulmonary lesion severity was negatively correlated with alveolar macrophage (AM) in both PRRS and PRDC groups on 14 and 28 dpi. AM in BALc was less populated in PRDC group on 28 dpi compared to PRRS group. AM in BALc was significantly less populated in PRDC group on 28 dpi compared to 14 dpi. In addition, cytotoxic T lymphocyte (CTL) in BALc was higher populated in PRDC group on 14 dpi and 28 dpi compared to PRRS group. In the case of PBMC cytokine TNF-α, IFN-α, IL-1β, IFN-γ, FoxP3, and IL-2, the PRRS group showed higher expression than the PRDC group on 7 dpi, 14 dpi, 7 dpi, 14 dpi, 14 dpi, and 14 dpi, respectively. On the other hand, in the case of IFN-β, IL-6, IL-8, IL-4, and IL-17, the PRDC group showed higher PBMC cytokine expression at 14 dpi, 7 dpi, 14 dpi, 3 dpi, and 3 dpi, respectively, than the PRRS group. Based on these results, our study could characterize differential immune responses in pigs with PRRS or PRDC.

Toxic Effect of Azalea Extract on Cardiovascular System (진달래 꽃잎의 추출물이 심혈관계에 미치는 영향)

  • Chun, Jun-Ha;Chung, Sung-Bok;Kang, Seung-Ho;Kim, Yeong-Jo;Shim, Bong-Sub;Lee, Hyun-Woo;Shin, Dong-Gu;Park, Jong-Min
    • Journal of Yeungnam Medical Science
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    • v.8 no.2
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    • pp.52-62
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    • 1991
  • The toxic effect of azalea extract, expecially on cardiovascular system, is relatively unclear. The purpose of this study is to study the possible underlying mechanism and effect of toxic ingredient of azalea on cardiovascular system. The 71 healthy rabbits were divided into 10 groups : In group as preliminary study ; 4cc of normal saline was administered intravenously(N) ; 0.7gm/kg and 1.0gm/kg of azalea extract was administered respectively in the same route, volume(A1, A2) ; atropine was administered intravenously(A) ; after pretreatment with atropine(0.04mg/kg) to block parasympathetic system, azalea extract was injected like the above groups(AA1, AA2) ; normal saline, 0.7gm/kg and 1.0gm/kg of azalea extract were administered respectively with 0.2cc(1 : 1000) epinephrine(E0,E1,E2). We measured the following indices at I minute interval during first 10 minutes and then 10 minute interval during next 30 minutes : RR interval, QTc interval, maximal systolic and diastolic pressure drop with occuring time and presence of significant arrhythmia. The results were as follows : 1. The changes of RR interval, QTc interval were significantly increased in groups by Azalea extract. The blood pressure change was significantly decreased in groups by Azalea extract. There were no significant differences according to dosage of Azalea extract. 2. The changes of RR interval, blood pressure were significant differences between administration of atropine and Azalea extract after pretreatment with atropine, but not in the change of QTc interval. 3. There were no significant differences in the change of RR interval, ATc interval, blood pressure drop according to pretreatment with atropine. 4. The interaction between epineprine and Azalea extract was not noted by the effect of epineprine itself. 5. The ST change by 0.7gm/kg, 1.0gm/kg of Azalea extract was revealed in 1 case(14.0%), 7 case(100%), respectively. 6. Most of all cases with arrhthymia, ventricular tachycardia, ventricular fibrillation, were noted in the group by epineprine, except one case by Azalea extract(1.0gm/kg). It was idioventricular rhythm. In conclusion, azalea extract has negative inotropic and chronotropic effect with arrhythmogenic potential possibly through direct myocardial ischemia or injury but we cann't be absolutely exclusive of actions of autonmic nervous system, especially parasympathetic nervous system.

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