• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.31 no.6
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• pp.461-467
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• 2005

Background. 5-(and-6)-carboxy-2',7'-dichlorofluorescein diacetate, succinimidyl ester mixed (CFSE) is the fluorescent labelling agent of living cells and used to trace the cells in vivo after transplatnation of various cells. The CFSE labelled cells can maintain fluorescence for up to 7 days after labelling. The MC3T3-E1 cell line (MC3T3) has been used for many studies about osteoblast, which is well known as a mouse preosteoblast. So the CFSE would be used to trace the transplanted MC3T3. However there are few reports about CFSE labelling of MC3T3. This study is aimed to know about adequate concenturation and incubation time of CFSE to MC3T3. Materials and methods. The MC3T3 was incubated in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ using ${\alpha}$-minimal essential medium (${alpha}$-MEM) containing10% FBS and gentamycin. Ten mM CFSE solution in dimethylsulphoxide (DMSO: 1%) was diluted with phosphate buffered saline (PBS) and final concentration of culture medium was, respectively, 5, 10, 15, 20, 25 and 30 ${{\mu}M$. Then the MC3T3 was incubated with CFSE in a humidified atmosphere of 95% air with 5% $CO_2$ at $37^{\circ}C$ for 5, 10, 15, 20, 25, 30, 35, 40 and 45 minutes in each concentration. The fluorescence of CFSE labelled cells was analysed with a inverted fluorescence microscope. The duration of cell labelling was also studied. Trypan blue dye exclusion test was done for cell viability. Results. For concentration between 5 and 10 ${\mu}M$, CFSE did not significantly label the MC3T3 in vitro. The destruction of MC3T3 was observed at the concentration of 20 ${\mu}M$. In the concentration of 15 ${\mu}M$, the best labelling was obtained at an incubation period between 15 and 30 minutes. The MC3T3 labelled with an incubation period of 15 minutes at 15 ${\mu}M$ was still fluorescent 7 days after CFSE labelling. The mean cell viability was 95.93%. Conclusion. These results suggests an incubation period of 15 minutes at 15 ${\mu}M$ of CFSE provides best labelling of MC3T3 in vitro.

• Journal of Physiology & Pathology in Korean Medicine
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• v.20 no.4
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• pp.936-941
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• 2006

To prevent human body injury from oxidative stress, antioxidants are very important and many research about antioxidants are generally being conducted. Hydrogen peroxide$(H_20_2)$ that is one of vitality oxygen species has been seen that cause various diseases, DNA damage and gene change. We have already known that the inhibition effect of Zizania latifolia Radix, Rhizoma on apoptosis induced by $H_2O_2$ in Neuro2A cell. And the purpose of this study was that we made a comparative study on the inhibition effect of apoptosis in Neuro2A cell on the region of Zizania latifolia(Radix, Rhizoma, Herba). Neuro2A cells were cultivated in RPMI(GibcoBRL) with 5% FBS and treated with $H_2O_2$ and Zizania latifolia(Radix, Rhizoma, Herba). Separately we measured the cell viability and analyzed DNA fragmentation. Activity of PARP, Cytochrome C, caspase-9, caspase-3, p53, p21, Bax and Bcl-2 in the cell was examined by using western blot. The results obtained were as Follows: The cell viability in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment (60ug/m1<) decreased significantly compared with that of none treatment(p<0.001). Zizania latifolia Radix increased cell viability was most effective of three regions. But we had no significant difference among three regions. All of Zizania latifolia (Radix, Rhizoma, Herba) increased cell viability about twice as much as that being injury by $H_2O_2$,(Zizania Latifolia (Radix, nhizoma, Herba) 20ug/m1, $H_2O_2$ 200uM, p<0.001). DNA fragmentation developed by $H_2O_2$, but was not developed in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. PARP, Cytochrome C, caspase-9 and caspase-3 activated all by $H_2O_2$ but were not activated in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. P53, P2l and Bax activated by $H_2O_2$, and Bcl-2 got into inactivation. But the opposite results appeared in all of Zizania latifolia (Radix, Rhizoma, Herba) treatment. In conclusion, these results suggest that all of Zizania latifolia (Radix, Rhizoma, Herba) inhibit the development of DNA fragmentation and apoptosis by $H_2O_2$and the antioridant action of all of Zizania latifolia (Radix, Rhizoma, Herba) is effective.

• The Journal of the Society of Stroke on Korean Medicine
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• v.8 no.1
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• pp.22-27
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• 2007

Objectives : This study was aimed to assess the distribution of metabolic syndrome according to Sasang constitution(SC) in acute stroke patients. Methods : We assessed the type of SC of acute stroke patients in the oriental medical hospitals of 3 universities located in metropolitan region from October 2005 to June 2007 by Ouestionnaire for the Sasang Constitution Classification II(QSCC II). Then, we investigated the general characteristics, the prevalence of each factor of the metabolic syndrome and the prevalence of the metabolic syndrome according to SC. Results : 338 subjects were included into the final analysis. 1. This study showed that the proportion of So-yang was highest of all constitutions(45.9%). 2. The prevalence of high TG, low HDL-chol, high BP, high FBS of Tae-um were higher than those of the other constitutions, but the differencecs were not statistically significant. The prevalence of high WC of Tae-um was significantly higher than other constitutions(p<0.001). 3. The prevalence of metabolic syndrome of Tae-um was significantly higher than other constitutions (p=0.002). Conclusion : There were significant differences in the prevalence of metabolic syndrome according to SC in acute stroke patients. For the further research, it seems to be necessary to construct fundamental databases for stroke by increasing the number of patients and by diagnosing SC more delicately.

• Development and Reproduction
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• v.17 no.3
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• pp.269-274
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• 2013

This study was performed to evaluate the effects of fibroblast co-culture on in vitro maturation (IVM) of prepubertal mouse preantral follicles. The intact preantral follicles were obtained from the ovaries of 12-14 day old mice and these were cultured individually in ${\alpha}$-minimal essential medium (${\alpha}$-MEM) supplemented with 5% fetal bovine serum (FBS), $100mIU/m{\ell}$ recombinant follicle stimulating hormone (rFSH), 1% insulin-transferrin-selenium, $100{\mu}g/ml$ penicillin and $50{\mu}g/m{\ell}$ streptomycin as base medium for 12 days. A total of 200 follicles were cultured in base medium co-cultured with mouse embryonic fibroblast (MEF) (MEF group) (n=100) or only base medium as control group (n=100). Survival rate of follicles on day 12 of culture were significantly higher in the MEF group of 90.0%, compared with 77.0% of the control group (p=0.021). Follicle diameters on day 6 and 8 of the culture period were significantly larger in the MEF group than those in the control group (p=0.021, p=0.007, respectively). Estradiol levels in culture media on day 4, 6, 8, 10 and 12 of the culture period were significantly higher in the MEF group (p=0.043, p=0.021, p=0.006, p<0.001 and p=0.008, retrospectively). Our data suggest that MEF cell co-culture on IVM of mouse preantral follicle increases survival rate and promotes follicular growth and steroid production.

• Nutrition Research and Practice
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• v.8 no.3
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• pp.336-341
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• 2014

BACKGROUND/OBJECTIVE: It is expected that dairy products such as cheeses, which are the main source of cholesterol and saturated fat, may lead to the development or increase the risk of cardiovascular and metabolic diseases; however, the results of different studies are inconsistent. This study was conducted to assess the association between cheese consumption and cardiovascular risk factors in an Iranian adult population. SUBJECTS/METHODS: Information from the Isfahan Healthy Heart Program (IHHP) was used for this cross-sectional study with a total of 1,752 participants (782 men and 970 women). Weight, height, waist and hip circumference measurement, as well as fasting blood samples were gathered and biochemical assessments were done. To evaluate the dietary intakes of participants a validated food frequency questionnaire, consists of 49 items, was completed by expert technicians. Consumption of cheese was classified as less than 7 times per week and 7-14 times per week. RESULTS: Higher consumption of cheese was associated with higher C-Reactive Protein (CRP), apolipoprotein A and high density lipoprotein cholesterol (HDL-C) level but not with fasting blood sugar (FBS), total cholesterol, low density lipoprotein cholesterol (LDL-C), triglyceride (TG) and apolipoprotein B. Higher consumption of cheese was positively associated with consumption of liquid and solid oil, grain, pulses, fruit, vegetable, meat and dairy, and negatively associated with Global Dietary Index. After control for other potential confounders the association between cheese intake and metabolic syndrome (OR: 0.81; 96%CI: 0.71-0.94), low HDL-C level (OR: 0.87; 96%CI: 0.79-0.96) and dyslipidemia (OR: 0.88; 96%CI: 0.79-0.98) became negatively significant. CONCLUSION: This study found an inverse association between the frequency of cheese intake and cardiovascular risk factors; however, further prospective studies are required to confirm the present results and to illustrate its mechanisms.

• Journal of Korean Medicine for Obesity Research
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• v.9 no.2
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• pp.33-45
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• 2009

Objectives This study was performed to evaluate the relation between hair tissue mineral elements(HTME) and Heart rate variability(HRV) of the metabolic syndrome(MS). Methods 89 persons(41-69 ages) who visited Oriental hospital for medical examination were divided MS group(n=22) and control group(n=67). HTME and HRV were compared, and were analyzed correlation with five contents of the MS. Results (1) In total subjects, waist circumference had a positive correlation with Pb. Blood pressure had a negative correlation with Mg. High density lipoprotein cholesterol(HDL-chol) had a negative correlation with Pb, Sb and K, while positive correlation with Ca/K. Triglyceride(TG) had a negative correlation with Mg and Na/K, while positive correlation with K and Ca/Mg. (2) In total subjects, TG and fasting blood sugar(FBS) had negative correlation with high frequency(HF), while positive correlation with low frequency/high frequency ratio(LF/HF). (3) In MS group, most of the level of toxic minerals were higher, and the level of major nutritional minerals were lower, but there were no statistical signficance. In two groups, there is no contrast between the correlations of the MS contents and HTME. (4) In MS group, HF was significantly lower and LF/HF was higher than normal group. LF/HF had a negative correlation with waist circumference in MS group, while positive correlation in normal group. (5) In total subjects, TP had a positive correlation with Mg. In two groups, there is no contrast between the correlations of the MS contents and HRV. However LF/HF had a positive correlation with Na in MS group, TP and LF had negative correlation with Ca, while RMSSD and HF had negative correlation with Cu in normal group. Conclusion These results may suggest that HTME and HRV are useful in diagnosing and preventing the metabolic syndrome.

• Journal of Korean Academy of Nursing
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• v.27 no.3
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• pp.563-576
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• 1997

This study was carried out to identify basic data for more efficient operation of comprehensive health check-up centers in the national health care system. The researcher reviewed and analyzed the main symptoms of the subjects according to sex, age, occupation and positive rate in the screening tests of a comprehensive health check-up, performed at one comprehensive health check-up center in located in a hospital. The subjects were 512 persons who had a check-up at the comprehensive health check-up, center in K university hospital in Seoul from October 2, 1996 to March 30, 1997. A questionaire developed by the researcher to obtain subject's general characteristics, main symptoms, medical examination, diagnosed disease was used as the tool for the study. The researcher analyzed the data with SPSS PC/sup +/ program. : the distribution of the subject's general characteristics and main symptoms was described with percentages and the difference between main symptoms and screening tests analyzed with X²-test, t-test, ANOVA. The result of this study are as follows : 1. For the distribution of diagnosis in males. liver disease was most frequent and hypertension was second : in females, hypertention and liver disease were not common in that order. 2. The most common complaints were indigestion in the digestive system, sputum in the respiratory system, irregular heart beat in the cardiovascular system, frequent urine in the urinary system vertigo in the hematologic system. 3. The main symptoms according to sex were for females, a statistically siguificant higher rate of complaints in the digestive system, cardiovascular system, urinary system, hematologic system, musculoskeletal, mental & nervous system as compared to males. 4. The main symptoms according to age were that in the the second decade there wara higher rate of complaints in the digestive system, and cardiovascular system than in the older age groups : and in the fifth decade more in the musculoskeletal system. 5. The main symptoms according to occupation were that formers and fishers had a higher rate of complaints in the mental and nervous system as compared to other occupation groups : and office workers, in gynecology. 6. The main symptoms according to diagnosis, were symptoms of the digestive system, frequent in liver disease : symptoms of the cardiovascular system in allergic disease. 7. The screening tests which showed the highest rates were UGI /FGS, PFT : In males a higher rate of abnormality was seen in abdominal ultrasono- graphy and in, females, in mammography. 8. In screening tests according to sex, males showed a higher positive rate for chest X-rays, and LFT, and females, in mammography, pap smear, CBC. 9. In screening tests according to age, higher positive rates of obesity, high blood pressure, chest X-ray, mammography, pap smear, FBS, CBC, urine test were seen in the fifth decade : PFT and abdominal ultrasonography in the third decade. The subjects who wanted comprehensive health check-up were not asymptomatic but had an illness with several symptoms. It is found that the most frequent diagnosed diseases were hypertention and liver disease. So proper observations must be done for them. Comprehensive health check-ups should be changed to a comprehensive examination including treatment beyond examination for those subjects with positive results.

• Journal of Periodontal and Implant Science
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• v.26 no.4
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• pp.841-858
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• 1996

Epidermal growth factor(EGF) is one of polypeptide growth factors. EGF has been reported as a biological mediator which regulates activities of wound healing process including the cell proliferation, migration and metabolism. The purposes of this study is to evaluate the effects of EGF on the human periodontal ligament cells and human gingival fibroblast cells that promote regeneration of periodntal tissue. The mitogenic effects of epidermal growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'-deoxy-uridine into DNA of the cells in a dose dependent manner. The prepared cells were the primary cultured gingival fibroblast and periodontal ligament cells from humans, the fourth or sixth subpassages were used in the experiments. Cells were seeded in DMEM containing 10% FBS. 1, 10, 50, 100, $200{\eta}g/ml$ and epidermal growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10\{mu}l/200{\mu}l$ 5-Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows : The DNA synthetic activity of human gingival fibroblasts were increased dose dependently by epidermal growth factor at 24 hours, 48 hours and 72 hours. The mitogenic effects were similar at the 24 and 48 hours of epidermal growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells were increased dose dependently by epidermal growth factor at 24 hours but the DNA synthetic activity decreased at $200{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were observed at the 48 hours application of epidermal growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 24, 72 hours than at 48 hours the application of epidermal growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the epidermal growth factor. In conclusion, epidermal growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

• The Journal of Korean Medicine
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• v.27 no.1 s.65
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• pp.165-183
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• 2006

Objectives : We implemented the Artemisia Extract Moxibustion Method and had the clinical tests for the diabetes with it. Methods : We implemented Artemisia extract made by extracting the vasodilator and antioxidant compounds from Artemisia-CH2C12 fraction and the moxibustion method constructed with DC Power supply, controller, Artemisia pad. single and multiple heating terminal with PTC(Positive Temperature Coefficient) thermistor. And we performed to estimate the efficiency on the questionnaire and the clinical tests with 23 cases of the diabetics. Results : We have estimated the improvement over 60% the symptoms that were the upper and lower limbs pain, frequent urination, spontaneous perspiration, thirst, decrease of body weight, and malaise after the moxibustion treatment on 5 cases among 23 cases. And the 19 cases took the biochemical check-up after the moxibustion treatment. From the results of biochemical check-up, the average HbAlc of before treatment was 8.400%, and after treatment 7.632%. The average HbAlc was decreased significantly after treatment (P<0.001). And the average urinary blood of before treatment was 0.73 and after treatment 0.27. The average urinary blood was decreased significantly after treatment (P<0.001). In addition the average FBS before treatment was 182.64 mg/dl, after treatment 161.77 mg/dl. Conclusions : We could estimate that our proposed moxibustion method was a significant treatment method for the diabetes.

• Journal of Embryo Transfer
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• v.19 no.3
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• pp.275-282
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• 2004

This study was carried out to examine the effects of development media and those change on in vitro development (IVD) of porcine oocytes matured and fertilized in vitro. Putative embryos, which were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF) and were fertilized in mTBM, were developed in vitro as the experimental scheme. The results are as follows. When porcine putative embryos were cultured in vitro in NCSU-23 or CZB/Pig-MEM, the percentage of oocytes cleaved was not different between two systems, but the percentage of blastocysts in NCSU-23 was significantly higher than in CZB/Pig-MEM (P<0.05). And when porcine putative embryos were cultured in vitro in NCSU-23 during 7 days with or without changing media at day 5, which was supplement with or without 10％ fetal bovine serum(FBS), the percentage of oocytes cleaved, blastocysts at day 6, and the cell number of ICM, TE and total of blastocysts at day 7 were not different among three treatments. As a result of this study, it is supposed that NCSU-23 be more favorable, to develop porcine embryos derived from IVM/IVF, than CZB/Pig-MEM, but that demonstration on the effects of changing medium with fresh one stand in need of the more experiments.