• Title/Summary/Keyword: alkaline phosphatase.

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Immunocytochemical Study on the Change of the Activated T Cells in Peripheral Blood of the Pulmonary Tuberculosis Patients (폐결핵환자의 말초혈액에서 Activated T Cell의 변화)

  • Ryu, Gyeong-Ryeol;Park, Une-Sook;Park, Chong-Hwa;Jeoung, Pan-Joon;Hwang, Young-Sil;Lee, Yang-Jae;Chang, Ga-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.6
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    • pp.823-830
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    • 1995
  • Background: It has been found that Helper T cells in the peripheral blood are decreased in the cell mediated immunity in the pulmonary tuberculosis. But it has not been confirmed yet that only decrease in number of cells which has phenotype in the peripheral blood is defined to decrease in cell mediated immunity. The immunocytochemical study was performed to observe the change of the percentage of T-lymphocytes with their subsets and activated T cells in the peripheral blood of pulmonary tuberculosis and to know how many T cells would be activated, relative to resting cells in the peripheral blood. Methods: The peripheral blood obtained from twenty two patients and ten healthy controls were smeared on the gelatin coated slide glass prepared for of mononuclear cells. The double bridge technique of alkaline phosphatase-antialkaline phosphatase(APAAP) method was used. As the primary antibodies, $T_1$(anti-human T cell), $T_4$(anti-human helper/inducer T cells) and $T_8$(anti-human supressor/cytotoxic T cell) antibodies and interleukin-2 receptor (for early activated T cell), very late activation antigen (for activated cytotoxic T cell), T cell lineage specific activation antigen monoclonal actibodies were used. Results: 1) There were significantly decrease in the absolute number of $T_4$(+) cells but significantly increase of $T_8$(+) cells in the peripheral blood of pulmonary tuberculosis (p<0.05). 2) The percentage of $T_4$(+) cells showed significantly decrease in pulmonary tuberculosis but $T_8$(+)cells significantly increase(p<0.05). $T_4(+)/T_8(+)$ ratio showed significantly decrease in the peripheral blood of the pulmonary tuberculosis(p<0.05). 3) There were significantly increase in the absolute number of variable stages of activated T cells in the peripheral blood of the pulmonary tuberculosis(p<0.05). 4) The percentage of IL-2R, VLA-1, TLiSA were 6.45+1.56%, $7.64+1.34^*$, 10.45+1.16% in order which showed significantly increase in the peripheral blood of the pulmonary tuberculosis(p<0.05). Conclusion: We speculate that only a few percentage of T lymphocyte is activated in cell mediated immunity in pulmonary tuberculosis.

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Effects of Ramie Leaves on Improvement of Lipid Metabolism and Antiobesity Effect in Rats Fed a High Fat/High Cholesterol Diet (모시잎이 고지방-고콜레스테롤 식이 흰쥐의 지질대사 개선 및 항비만 효과에 미치는 영향)

  • Lee, Jae-Joon;Park, Mi-Ran;Kim, Ah-Ra;Lee, Myung-Yul
    • Korean Journal of Food Science and Technology
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    • v.43 no.1
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    • pp.83-90
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    • 2011
  • This study was performed to investigate the effects of ramie (Boehmeria nivea) leaf powder on improvements in lipid metabolism in serum, liver and adipose tissue, and the anti-obesity effect in rats fed a high fat/high cholesterol diet for 4 weeks to induce a hyperlipidemic and obese model rat. Male Sprague-Dawley rats weighting 210 g were divided into four groups; a normal diet group (N), a high fat/high cholesterol diet group (HFC), a high fat/high cholesterol diet with 5% ramie leaf powder group (HFC-RL), and a high fat/high cholesterol diet with 10% ramie leaf powder group (HFC-RH). The body weight gain increased with a high fat/high cholesterol diet, but gradually decreased in the ramie leaf powder fed groups compared with the HFC group. The liver index in the HFC group was highest among the four groups, although the difference was not significant compared with the ramie leaf powder fed groups. The adipose tissues weight in the HFC group was heavier than that of the N group, whereas those of groups fed ramie leaf powder decreased gradually. Alkaline phosphatase activity was not different between the HFC groups, but serum alkaline aminotransferase and lactate dehydrogenase activities decreased significantly after ramie leaf powder feeding. Levels of serum triglyceride, total cholesterol, LDL-cholesterol, atherogenic index and cardiac risk factors tended to decrease in the ramie leaf powder fed groups compared with the HFC group, whereas serum HDL-cholesterol level decreased in the HFC group and markedly increased in the ramie leaf powder fed groups. Levels of triglyceride and total cholesterol in the liver were significantly lower in the ramie leaf powder fed groups than in the HFC group. Levels of triglyceride and total cholesterol in adipose tissues were also lower in the ramie leaves powder fed groups than in the HFC group. The activities of heparinreleasable lipoprotein lipase (LPL) and total-extractable LPL in adipose tissues increased in the HFC group compared to that of the N group, but those of the ramie leaf powder fed groups decreased significantly. These results suggest that ramie leaf powder may improve lipid metabolism in serum, liver and adipose tissue and potentially reduce lipid storage.

Effects of High Pressure Treatment on the Microbiological and Chemical Properties of Milk (초고압 처리가 우유의 미생물학적 및 이화학적 특성에 미치는 영향)

  • Lee, Jieun;Choi, Eun-Ji;Park, Sun Young;Jeon, Ga Young;Jang, Ja-Young;Oh, Young Jun;Lim, Seul Ki;Kim, Tae-Woon;Lee, Jong-Hee;Park, Hae Woong;Kim, Hyun Ju;Jeon, Jung Tae;Choi, Hak-Jong
    • Microbiology and Biotechnology Letters
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    • v.42 no.3
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    • pp.267-274
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    • 2014
  • High pressure processing (HPP) is a non-thermal method used to prevent bacterial growth in the food industry. Currently, pasteurization is the most common method in use for most milk processing, but this has the disadvantage that it leads to changes in the milk's nutritional and chemical properties. Therefore, the effects of HPP treatment on the microbiological and chemical properties of milk were investigated in this study. With the treatment of HPP at 600 MPa and $15^{\circ}C$ for 3 min, the quantity of microorganisms and lactic acid bacteria were reduced to the level of 2-3 log CFU/ml, and coliforms were not detected during a storage period of 15 d at $4^{\circ}C$. An analysis of milk proteins, such as ${\alpha}$-casein, ${\beta}$-casein, ${\kappa}$-casein, ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin by on-chip electorophoresis revealed that the electrophoretic pattern of the proteins from HPP-treated milk was different from that of conventionally treated commercial milk. While the quantities of vitamins and minerals in HPP-treated milk were seen to be comparable to amounts found in raw milk, the enzyme activity of lipase, protease and alkaline phosphatase after HPP treatment was reduced. These results suggest that HPP treatment is a viable method for the control of undesirable microorganisms in milk, allowing for minimal nutritional and chemical changes in the milk during the process.

Lethal Toxicity and Hematological Changes Exposed to Nitrate in Flatfish, Paralichthys olivaceus in Biofloc and Seawater (바이오플락 및 일반 해수에서 질산염의 넙치 (Paralichthys olivaceus)에 미치는 급성 독성 및 혈액학적 성상의 변화)

  • Bae, Sun-Hye;Kim, Ki Wook;Kim, Su Kyoung;Kim, Su-Kyoung;Kim, Jong-Hyun;Kim, Jun-Hwan
    • Korean Journal of Environmental Biology
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    • v.35 no.3
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    • pp.373-379
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    • 2017
  • Juvenile Paralichthys olivaceus (mean length $19.8{\pm}2.6cm$, mean weight $97.8{\pm}15.8g$) were exposed for 96 hours to different nitrate concentrations of 0, 62.5, 125, 250, 500, 1,000, and $1,500mg\;L^{-1}$ in biofloc and 0, 62.5, 125, 250, 500, and $1,000mg\;L^{-1}$ in seawater. Median lethal concentration values ($LC_{50}$, the concentration at which 50% of mortality occurred after 96 hours of exposure) of nitrate to P. olivaceus in biofloc and seawater were 1,226 and $597mg\;NO_3L^{-1}$ (P<0.05), respectively, revealing a higher toxicity of nitrate to P. olivaceus in seawater than in biofloc. In hematological parameters, hematocrit level in P. olivaceus exposed to nitrate was significantly increased only at a concentration of $1,000mg\;L^{-1}$ in biofloc and at concentrations higher than $250mg\;L^{-1}$ in seawater, but no significant changes in hemoglobin were found in biofloc and seawater. In plasma parameters, aspartate aminotransferase (AST) and alanine aminotransminase (ALT) were significantly increased by nitrate exposure in biofloc and seawater, but no significant changes in alkaline phosphatase (ALP) were found in biofloc and seawater. Results of this study indicate that nitrate exposure to P. olivaceus have a lethal toxic effect and alter hematological and plasma constituents of flatfish P. olivaceus. Given relatively lower toxicity of nitrate in biofloc than in seawater, the use of biofloc in aquaculture may reduce potential toxic effect caused by nitrate in feces and feed residue.

Effects of Green Tea Polyphenol and Vitamin C on Type 2 Diabetic Rats Induced by Low Dose Streptozotocin Following High Fat Diet (고지방식이와 저용량 스트렙토조토신으로 유도된 2형 당뇨병 흰쥐에서 녹차 폴리페놀과 비타민 C 병합 투여 효과)

  • Lee, Byoung-Rai;Yang, Hoon;Park, Pyoung-Sim
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.2
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    • pp.167-173
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    • 2016
  • This study investigated the effects of green tea polyphenol and vitamin C on type 2 diabetes mellitus by administering polyphenon 60 (P60) and sodium ascorbate (SA) to diabetic rats induced by high fat diet/low-dose streptozotocin. The experimental group was divided into five different groups: non-diabetic control group (NC), diabetes control group (DC), diabetes+P60 group (DM+P60), diabetes+SA group (DM+SA), and diabetes+P60+SA group (DM+P60+SA). P60 and SA were dissolved in 0.1% drinking water. After P60 and SA were administered for 16 weeks, fasting blood glucose, plasma insulin, serum triglyceride, blood urea nitrogen (BUN), and creatinine levels as well as kidney alkaline phosphatase (AP) and ${\gamma}$-glutamyltranspeptidase (GGT) activities were measured. Fasting blood glucose level increased 5-fold in the DC group compared to the NC group. In the DM+P60 group, fasting blood glucose level decreased by 14%. In the DM+P60+SA group, fasting blood glucose level decreased by 28% compared to the DC group, whereas the DM+SA group did not show any significant difference. The homeostasis model assessment for insulin resistance index increased in the DC group and decreased in the DM+P60+SA group compared to the DC group. Serum creatinine level increased in the DC group, but decreased by 17% in the DM+P60 group and by 43% in the DM+P60+SA group compared to the DC group. The serum BUN level increased in the DC group, but decreased by 41% in the DM+P60+SA group compared to the DC group. Kidney GGT and AP activities decreased in the DC group compared to the NC group; however, they were reversed by DM+P60+SA group. These results show that combined administration of both green tea polyphenol and vitamin C had better effects on improving blood glucose level, insulin resistance, serum triglyceride level, and protecting kidneys than administration of either green tea polyphenol or vitamin C alone in the context of type 2 diabetes.

A STUDY ABOUT ALVEOLAR CREST BONE HEIGHT BEFORE AND AFTER ORTHODONTIC TREATMENT BY USING BITEWING FILM (교익사진을 이용한 교정치료 전후의 치조골 높이 변화에 관한 연구)

  • Hwang, Chung-Ju
    • The korean journal of orthodontics
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    • v.27 no.3 s.62
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    • pp.421-430
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    • 1997
  • Alveolar bone grows with development of tooth germs and roots; bone deposition occurs with tooth eruption. Bone components undergoes processes of resorption and deposition, and when the balance between them is disrupted, decrease in alveolar bone height or excessive bone deposition result. It has been hon that repositioning of teeth through orthodontic treatment can cause alveolar bone resorption which result in decreased alveolar bone height, and there have been many studies to evaluate such effects. X-ray films that could be replicated and standardized were chosen in clinical studies, and among them, bitewing films were used for objective evaluation of changes in alveolar bone level. Twenty subjects, 10 to 13-year- old (average 12.2) children with Cl I molar key, healthy oral condition, no congenital missing, no periodontal disease, and pre-and post-orthodontic bitewing films, were randomly selected for comparison of alveolar bone heights. Amounts of tooth and changes in alveolar bone heights were analyzed. The following results were obtained: 1. Amount of tooth movement in canine, premolar, and molar regions, changes in tooth axis, and changes in alveolar bone heights were measured, and the mean and median values were obtained. 2. When pre-and post-orthodontic alveolar bone levels were compared, larger changes were noticed in maxilla than mandible. 3. When mesio-distally compared, larger changes were observed in the distal sides of 3D3 and 4M3, mesial sides of 4M3 and 4D3, distal sides of 4D3 and 5M3, mesial sides of 5M3 and 5D3, md distal sides of 5D3 and 6M3. 4. When the amounts of tooth movements(TX, TY)and changes in tooth axis(A) were compared,34TX, 34TY, 34A of both sides in maxilla were greater, iud changes in alveolar bone level were greater than any other region.

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Effects of Radiofrequency Induced Local Hyperthermia on Normal Canine Liver (라디오 파를 이용한 국소 온열치료 : 정상 개의 간에 미치는 영향)

  • Suh Chang Ok;Loh. John J.K.;Shin Hyun Soo;Lee Hyung Sik;Moon Sun Rock;Seong Jin Sil;Chu Sung Sil;Kim Gwi Eon;Han Eun Kyung;Park Chan Il
    • Radiation Oncology Journal
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    • v.9 no.1
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    • pp.37-45
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    • 1991
  • In order to assess the effects of radiofrequency-induced local hyperthermia on the normal liver, histopathologic findings and biochemical changes after localized hyperthermia in canine liver were studied. Hyperthermia was externally adminsitered using the Thermotron RF-8 (Yamamoto Vinyter Co., Japan; Capacitive type heating machine) with parallel opposed electrodes. Thirteen dogs were used and allocated into one control group (N=3) and two treatment groups according to the treatment temperature. Group I (N=5) was heated with $42.5\pm0.5^{\circ}C$ 30 minutes, and Group II (N=5) was heated with $45\pm0.5^{\circ}C$ for 15-30 minutes. Samples of liver tissue were obtained through a needle biopsy immediately after hyperthermia and T,14, and 28 days after treatment. Blood samples were obtained before treatment and W, 3,5, 7,14 and 28 days after treatment and examined for SGOT, SGPT and alkaline phosphatase. Although SGOT and SGPT were elevated after hyperthermia in both groups (three of five in each group), there was no liver cell necrosis or hyperthermia related mortality in Group 1. A hydropic swelling of hepatocytes was prominent histologic finding. Hyperthermia with $45^{\circ}C$ for 30 minutes was fatal and showed extensive liver cell necrosis. In conclusion, liverdamage dy heat of $42.5\pm0.5^{\circ}C$ for 30 minutes is reversible, and liver damage by heat of $45\pm0.5^{\circ}C$ for 30 minutes can be fatal or irreversible. However, these results cannot be applied directly to human trial. Therefore, in erder to apply hyperthermic treatment on human liver tumor safely, close obsewation of temperature with proper thermometry is mandatory. Hyperthermic treatment should be confined to the tumor area while sparing a normal liver as much as possible.

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Clinicopathological Effects of Waterpepper (Persicaria hydropiper) on Ruminants (여뀌섭취가 반추수에 비치는 임상병리학적 영향)

  • Cho Myoung-Rae;Han Hong-Ryul
    • Journal of Veterinary Clinics
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    • v.6 no.2
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    • pp.227-259
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    • 1989
  • Waterpepper is a weed which grows on damp soil. especially near swamps, and in shallow water of ponds and ditches. It Is widespread throughout the country In abundant colonies. In the present experiments, possible toxic effects of waterpepper were investigated in ruminants. Pour cows were fed waterpepper ad libitum or by force in the from of green forage, hay and/or powder, 8 goats were administered in the form of methanol extract, and 4 goats, crude juice, into the lumen. Clinical signs were examined as well as urinalysis, hematology, serum chemical analysis, pH/blood gas analysis and chclinesterase activities following administration of waterpepper. Six goats which were administered the methanol extract or crude juice were sacrificed for pathological examinations., In addition to the clini copathological examinations, the chemical constituents of waterpepper were qualitively analyzed from the methanol extract and the Effects of the waterpepper crude juice were examined on the motility of rabbit duodenum and uterus. It is revealed that waterpepper contains steroids, terpenoids, flavonoids, tannin and essential oils in the methanol extract and nitrates in the crude juice. The crude juice of waterpepper relaxed the rabbit uterine and duodenal smooth muscles. The constraction of duodenum by acetylcholine or BaCl$_2$ were partially inhibited by pretreatment of the crude juice. However, the relaxation of duodenum by the crude juice was not blocked by the pretreatments of phenoxybenzamlne, propranolol, cocaine, reserpine and tetrodotoxin. The constituents of waterpepper to evoke elaxation of duodenal smooth muscle were stable to heat. The cows administered waterpepper showed common clinical symptoms such as acrid expression, restlessness, dullness, inappetence, anorexia, severe diarrhea, mild bloat and left displacement of abomasum, while bloody feces was shown in a cow. The goats administered the mothanol extract showed common clinical signs such as acrid expression, restlessness, dullness, inappetence and soft feces, while bloody feces was shown in a goat, A goat adminstered the crude juice showde bloody feces and diarrhea. Respiratory rates and heart beats were increased along with diarrhea in the experimental cows. The erythrocyte counts and MCHC were decreased whereas PCV, MCV and neutrophils were increased in the cows administered waterpepper. In goats administered methanol extract, there were decreases in erythrocytes, PCV and hemoglobin content, and an increase in MCHC. The goats ingester with the crude juice showed negligible changes in hematologic values compared with control group which was administered the same amount of water instead of the crude juice. The contents of serum calcium, Inorganic phosphorus, magnesium, Iron, glucose, cholesterol, total protein, triglycerides and phospholipids were tended to decrease in cows. In goats serum iron, glucose, triglycerides, cholesterol, BUN and phospholipids content were decreased while the content of sodium and chloride were increased after administration of the methamol extract The goats ingested with the crude juice did not show significant changes in serum chemical analysis. Even though there were some pathological findings such as hyperemia in the small intestines and kidneys and swelling of liver parenchymal cells, the values of serum AST, ALT, LDH, alkaline phosphatase, total bilirubin and creatinine did not change significantly. While proteins, hemoglobin and blood were detected in the urine of cows, urine pH, ketone bodies, glucose, bilirubin and urobilinogen content were normal or undetected. There were no significant changes in pH/bolld gas analysis data of cows and cholinesterase activities of plasma and erythrocytes of cows and goats ingested with waterpepper or the methanol extract. It is concluded that waterpepper irritates the gatrointestinal system, causes abdominal pain, relaxes the gastrointestinal smooth muscle and dilatates blood vessels supplied to the system. The irritation and relaxation may lead to abnormal fermentation, maldigestion and malabsorption of nutrients and result in diarrhea, body feces, mild bloat and left displacement of abomasum.

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Effects of Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) on Osteoporosis Induced by Ovariectomy in Rats (유향흑호단(乳香黑虎丹)이 난소적출로 유발된 흰쥐의 골다공증 모델에 미치는 영향)

  • Kum, Chang-Jun;Cho, Chang-Young;Lim, Jeong-Min;Song, Dea-Uk;Kim, Jae-Hyun;Lee, Bina;Jung, Hyuk-Sang
    • Journal of Korean Medicine Rehabilitation
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    • v.24 no.3
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    • pp.29-37
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    • 2014
  • Objectives Osteoporosis is an impending problem in the rapidly aging society. The purpose of this study is to identify the effects of Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) on osteoporosis induced by ovariectomy in rats. Methods 24 rats were randomly assigned to a SHAM group, a control group, and a Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group (n=8). Ovaries were extracted and for 8 weeks, the rats were given dry feeds and Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group were given a mixture of dry feeds and Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$). At 8 weeks, their body weight, femur mass, tibia ash per body ratio, overall surface area and thickness of the trabeculae, overall surface area of the osteoblasts, and the number of osteoclasts were measured and levels of albumin, aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total-cholesterol, calcium and estradiol were recorded. Results There was no significant difference in the weight of the femur and the tibia ash per body ratio. The Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group had significantly thicker trabeculae than the control group and showed a minimal increase of overall surface area of the trabeculae. The overall surface area of the osteoblasts and the number of osteoclasts decreased in the Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group. There were no statistically significant differences in AST, ALT, ALP, total cholesterol, phosphorus, and estradiol levels. On the contrary the Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) group had significantly higher levels of Albumin, triglyceride, and calcium. Conclusions It can be concluded that oral intake of Yoohyangheukho-dan ($R{\check{u}}xi{\bar{a}}ngh{\bar{e}}ih{\check{u}}-d{\bar{a}}n$) prevents the thinning process of the trabeculae. Thus, it may have positive effects on the treatment and prevention of osteoporosis in postmenopausal women.

The effect of admixture of vitamin $D_3$ and dexamethasone on the activity of osteoblastic cells (Vitamin $D_3$와 Dexamethasone의 복합 투여가 골모세포에 미치는 영향에 관한 연구)

  • Lim, Na-Won;Park, Young Joo;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.29 no.3 s.74
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    • pp.383-397
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    • 1999
  • Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine $D_3$ are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucocorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin $D_3$, 1,25-dihydroxycholecalciferol[1.25-$(OH)_2D_3$], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin $D_3$ and dexamethasone, one of glucocorticoids, on osteoblastic cell line(MC3T3-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nM/ml of 1,25-$(OH)_2D_3$ and/or 10nM/ml, 100nM/ml, $1{\mu}M/ml$ of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with $1{\mu}M/ml$ of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in $1{\mu}M/ml$ of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin $D_3$ was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in l00nM/.ml at 2-day cultivation, and was a little increased again at 3-day cultivation. The activity of ALP was increased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin $D_3$, the cellular activity was decreased in any concentration of vitamin $D_3$ at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin $D_3$ at 2-day cultivation, and was again decreased at 3-day cultivation.

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