• Title/Summary/Keyword: In vivo와 In vitro

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The Ability of Cervus Elaphus Sibiricus Herbal Acupuncture to Inhibit the Generation of Inflammatory Enzymes on Collagen-induced Arthritis Mice (녹용약침(鹿茸藥鍼)이 CIA 모델 생쥐의 염증인자 생성억제에 미치는 영향)

  • Hwang, Jong-Soon;Hwang, Ji-Hye;Lee, Hyun-Jin;Lee, Dong-Gun;Kang, Min-Joo;Back, Song-Ook;Cho, Hyun-Seok;Kim, Kyung-Ho;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.24 no.6
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    • pp.1-14
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    • 2007
  • Backgrounds : Rheumatoid Arthritis(RA) is known as the chronic inflammatory diseasethat induces persistent inflammation in the joint cavity. The destruction of cartilage occurs as the result of bones destoyed by pannus, several influential cytokines induced by the synovial capsulitis, varieties of proteinases, $O_2$ radicals, and the secondary degenerative changes of articular cartilage. The type 2 collagen-induced arthritis model is used in recent experimental research on rheumatoid arthritis. Cervus elaphus sibiricus (Nockyong) has the effect of relieving pain by nourishing the muscles, joints, and bones. It is also known to be efficacious in promoting and enhancing the immune system. The objective of this study was to investigate the effect of Cervus elaphus sibiricus herbal acupuncture to inhibit the generation of proinflammatory enzyme on type 2 collagen-induced arthritis. I investigated the inhibition of mRNA transcription of MIF(macrophage migration inhibitory factor), $TNF-{\alpha}$(Tumor necrosis $factor-{\alpha}$) and MMP-9 (matrix metalloproteinase-9) of Cervus elaphus sibiricus herbal acupuncture using an in vitro test. Also investigated was the inhibition of differentiation of Th 1 cells and activation of cytokines(MIF, $TNF-{\alpha}$, IL-6, MMP-9), which are known to cause initial RA ,and are also related to the morphology of the synovial membranes of the joint capsule, by an in vivo test, using CIA(collagen induced arthritis) model mice. Materials & methods : The laboratory animals used in this experiment were 4 week-old DBA female mice, weighing approximately 20 grams, and adjusted to the laboratory environment. The experiment was divided into the normal group(NOR)-no treated group, control group(CON)-CIA induced group, and sample group(SAM)-Cervus elaphus sibiricus herbal acupuncture treated group. RA was induced in the mice via injection of $50{\mu}{\ell}$ C II mixed CFA. The Cervus elaphus sibiricus herbal acupuncture solution was applied on $GB_{35}$(陽陵泉) for 26 days from the 3rd day of RA inducement. The concentration of the solution was determined via a MTT assay. To research the effect on the expression of MIF, $TNF-{\alpha}$ and MMP-9 mRNA, RT-PCR was performed on synovial membrane cells from the knee joint of CIA mice. C II induced RA knee joint's histo-chemical synovial membrane was observed using a specimen model via the Hematoxilin and Eosin dying technique. Results : The expression of mRNA of RA-related cytokines such as MIF, $TNF-{\alpha}$, and MMP-9 dosedependently decreased in the cell from the synovial membranes of the joint, which is treated with Cervus elaphus sibiricus herbal acupuncture solution. In mice treated with Cervus elaphus sibiricusherbal acupuncture, the damage of synovial membranes of the joint was lessened, and differentiation of Th 1 cells was suppressed. The activation of RA-related cytokines such as MIF was suppressed, and the generation of $TNF-{\alpha}$ and MMP-9 showed a statistically significant decreas. Conclusions : It is speculated that Cervus elaphus sibiricus herbal acupuncture has the therapeutic effect of palliating the damage of the tissue impaired by RA by inhibition of the initial RA progression and by regulating excessive differentiation of Th 1 cell as it suppresses the generation of RA-related cytokines during the highest stage of RA by acting on pro-inflammatory enzymes.

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Effects of Herbal Complex on Blood Glucose in Streptozotocin-induced Diabetic Rats and in Mice Model of Metabolic Syndrome (생약복합제의 Streptozotocin 유발 당뇨 및 대사성증후군 모델 동물에서의 혈당에 미치는 효과)

  • Park, Han-Seok;Lee, Yeon-Sil;Choi, Se-Jin;Kim, Jin-Kyu;Lee, Yun-Lyul;Kim, Hyun-Gwen;Koo, Sam-Hoi;Ku, Dae-Hoy;Ki, Seung-Il;Lim, Soon-Sung
    • Korean Journal of Pharmacognosy
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    • v.40 no.3
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    • pp.196-204
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    • 2009
  • This study was carried out to investigate the in vivo and in vitro inhibitory effect of a traditional herbal complex (HC) extract prepared from a mixture of four oriental herbs (Dioscorea Rhizoma, Glycine soja Sieb. et Zucc, Bombycis corpus, Fermented Glycine soja) that have been widely used for the treatment and prevention of diabetes mellitus on hyperglycemia. The water extract of HC showed potent inhibitory effect on $\alpha$-glucosidase with $IC_{50}$ value of 1.24 mg/mL. Additionally, the ethanol extract of HC was also found to exhibit significant inhibitory effect against protein tyrosine phosphatase $1{\beta}$ ($PTP1{\beta}$), which is known as a major regulator of both insulin and leptin signaling. In the $PTP1{\beta}$ inhibitory assay, the most active n-hexane fraction obtained from the ethanol extract of HC, was identified as a mixture of fatty acid derivatives by gas chromatography-mass spectrometry (GC-MS). In high-fat diet-low dose streptozotocin (STZ)-induced diabetic rat, the water extract of HC improved the oral glucose intolerance as compared with rosiglitazone. HC also caused a marked decrease of body weight and fasting blood glucose and a significant improvement on glucose tolerance in metabolic syndrome mice model. These findings support that this traditional HC may be useful in the control of blood glucose in diabetes mellitus and metabolic syndrome.

Identification and Characterization of Lactobacillus salivarius subsp. salivarius from Korean Feces

  • Bae, Hyoung-Churl
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2004.05a
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    • pp.89-119
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    • 2004
  • This study was conducted to isolate lactobacilli having probiotic characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at $37^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by $0.9{\sim}1.0%$ at $37^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

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Lipase-Inhibitory and Anti-Oxidative Activity of the Methanol Extract and the Powder of Phellinus linteus (상황버섯 자실체 메탄올 추출물과 분말의 지방소화효소 억제 및 항산화 활성)

  • Kim, Ji-Hyun;Son, In-Suk;Kim, Jong-Sang;Kim, Ki-Hoon;Kwon, Chong-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.2
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    • pp.154-161
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    • 2008
  • Phellinus linteus (PL) has been known to exhibit potent biological activity. The present study was designed to investigate lipase-inhibitory and anti-oxidative activity of the methanol extract and the powder of PL fruiting body. The methanol extract of PL appeared to have the inhibitory activity against pancreatic lipase with an $IC_{50}$ value of $36.3\;{\mu}g/mL$, and the scavenging activity of DPPH radical with an $IC_{50}$ value of $20.1\;{\mu}g/mL$, which was similar to that of vitamin C ($IC_{50}\;18.3\;{\mu}g/mL$). To investigate the lipase-inhibitory and anti-oxidative effect of PL on animal, Sprague-Dawley rats were fed with high-fat diet supplemented with either 2% or 5% PL powder for 8 weeks. Total food intake was significantly increased, but body weight was not changed by PL powder supplementation. However, fecal fat excretion of the experimental groups fed with the PL powder were higher than that of the control group. PL powder showed a decrease in the plasma total cholesterol, LDL-cholesterol, and the hepatic total cholesterol levels. The anti-oxidative enzyme activities were also affected by PL supplementation. Glutathione peroxidase (GSH-Px) in the plasma and liver were significantly increased by 98% and 46% in the 2% PL group, and 99% and 32% in the 5% PL group, respectively. Total superoxide dismutase (T-SOD) activity was not affected by PL supplementation. DNA damage was measured by the comet assay in the lymphocytes collected after 2 weeks, 4 weeks, and 8 weeks of feeding PL supplemented diet. Lymphocyte DNA damage was decreased in the PL supplemented group. Furthermore, PL feeding enhanced the resistance to lymphocyte DNA damage caused by an oxidant challenge with $H_2O_2$.

Effect of Hijikia fusiforme Fractions on Proliferation and Differentiation in Osteoblastic MC3T3-E1 Cells (톳 분획물이 조골세포의 증식 및 분화에 미치는 영향)

  • Jeon, Min-Hee;Kim, Mi-Hyang
    • Journal of Life Science
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    • v.21 no.2
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    • pp.300-308
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    • 2011
  • Osteoporosis is a disease involving a decrease in bone mineral density and increased risk of fractures. Osteoblast and osteoclast activities are important for bone formation. The MC3T3-E1 osteoblastic cell line is a well-accepted model of osteogellsis in vitro. Hijikia fusiforme is a kind of edible brown seaweed that grows mainly in the Northwest Pacific region, including the countries of Korea, Japan and China, and it has been widely used as a medicinal and health food in Korea. In this study, by using osteoblasts, the effects of Hijikia fusiforme fractions on proliferation, alkaline phosphatase (ALP) activity, collagen synthesis and mineralization of cells were investigated. Hijikia fusiforme were subjected to fractionation by using hexane, methanol, butanol and aqueous. Proliferation of the MC3T3-E1 osteoblastic cells that were treated with Hijikia fusiforme fractions increased by approximately 120%. Regarding effects of Hijikia fusiforme fractions on ALP activity, 1 ${\mu}g$/ml butanol fraction showed the highest activity. The synthesis of collagen increased significantly in response to treatment with Hijikia fusiforme fractions, with the exception of the hexane fraction. Moreover, mineralization in the MC3T3-E1 cells that were treated with 100 ${\mu}g$/ml butanol fraction increased by 281%. Also, when 100 ${\mu}g$/ml aqueous fraction was added, mineralization increased by 240%. These results indicate that Hijikia fusiforme fractions have anabolic effect on bone through the promotion of osteoblastic differentiation, suggesting that it could be used for the treatment of common metabolic bone diseases.

Measurement of Porcine Aortic and Pulmonary Valve Geometry and Design for Implantable Tissue Valve (돼지 대동맥, 폐동맥의 근위부 기하학적 구조 측정을 통한 판막 구조 수치의 계량화와 판막 도안에 관한 연구)

  • Park, Sung-Joon;Kim, Yong-Jin;Nam, Jin-Hae;Kim, Soo-Hwan;Lee, Chang-Ha;Lim, Hong-Gook
    • Journal of Chest Surgery
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    • v.43 no.6
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    • pp.602-613
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    • 2010
  • Background: As life expectancy has been increased, the cardiac valve disease has been increased. In past, mechanical valve for valve replacement surgery was used widely, but it has many weaknesses, such as hemorrhage, teratogenic effect caused by warfarin, acute mechanical failure, taking warfarin during life, etc. So, the tissue valve is used widely and researches for durability of tissue valve are in progress. Tissue valves being used are all imported in Korea, and there is a lack of information on its geometry and design. So, we studied the geometry of porcine aortic and pulmonary valve, and tried to suggest theoretical basis for making the aortic and pulmonary valve. Material and Method: We harvested aortic and pulmonary valves of 25 pigs and measured the geometry of valve at fresh and glutaraldehyde (GA) fixed state. In each group, we measured the diameter of the base, diameter of commissure, valve height, commissural height, etc. Also, for making implantable porcine and bovine pericardial valve, we designed the valve stent form, thickness, height, and leaflet size, form, thickness by different size of valve. Result: The aortic and pulmonary valve geometry and ratio were measured in each group. The right coronary cusp of aortic valve and right facing cusp of pulmonary valve was bigger than other cusps and non coronary cusp was smaller than others (RCC: NCC : LCC=1 : 0.88 : 1). Valve height was correlated to the leaflet size. We designed the outer diameter of stented porcine aortic valve from 19 mm to 33 mm and designed stent height and width, using previous measured ratio of each structure, stent thickness, working thickness (for making valve). Also, we designed the size of stent and form for stented bovine pericardial valve, considering diameter of valve, leaflet length, height and leaflet minimum coaptation area. Conclusion: By measuring of 25 pig's aortic and pulmonary valve geometry and ratio, we can make theoretical basis for making implantable stented porcine valve and bovine pericardial valve in various size. After making implantable valve using these data, it is necessary to do in vivo and in vitro researches, furthermore.

Biocompatibility and Histopathologic Change of the Acellular Xenogenic Pulmonary Valved Conduit Grafted in the Right Ventricular Outflow Tract (우심실 유출로에 이식한 무세포화 이종 폐동맥 판막도관의 생체 적합성 및 조직병리학적 변화양상에 대한 연구)

  • 허재학;김용진;박현정;김원곤
    • Journal of Chest Surgery
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    • v.37 no.6
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    • pp.482-491
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    • 2004
  • Background: The xenogenic or allogenic valves after in Vitro repopulation with autologous cells or in vivo repo-pulation after acellularization treatment to remove the antigenicity could used as an alternative to synthetic polymer scaffold. In the present study, we evaluated the process of repopulation by recipient cell to the acellu-larized xenograft treated with NaCl-SDS solution and grafted in the right ventricular outflow tract. Material and Method: Porcine pulmonary valved conduit were treated with. NaCl-SDS solution to make the grafts acellularized and implanted in the right ventricular outflow tract of the goats under cardiopulmonary bypass. After evaluating the functions of pulmonary valves by echocardiography, goats were sacrificed at 1 week, 1 month, 3 months, 6 months, and 12 months after implantation, respectively. After retrieving the implanted valved conduits, histopathologic examination with Hematoxylin-Eosin, Masson' trichrome staining and immunohistochemical staining was performed. Result: Among the six goats, which had been implanted with acellularized pulmonary valved conduits, five survived the expected time period. Echocardiographic examinations for pulmonary valves revealed good function except mild regurgitation and stenosis. Microscopic analysis of the leaflets showed progressive cellular in-growth, composed of fibroblasts, myofibroblasts, and endothelial cells, into the acellularized leaflets over time. Severe inflammatory respon-se was detected in early phase, though it gradually decreased afterwards. The extracellular matrices were regenerated by repopulated cells on the recellularized portion of the acellularized leaflet. Conclusion: The acellularized xenogenic pulmonary valved conuits were repopulated with fibroblasts, myofibroblasts, and endothelial cells of the recipient and extracellullar matrices were regenerated by repopulted cells 12 months after the implantation. The functional integrity of pulmonary valves was well preserved. This study showed that the acellularized porcine xenogenic valved conduits could be used as an ideal valve prosthesis with long term durability.

In Vivo and In Vitro Studies of the Steady State Free Precession-Diffusion-Weighted MR Imagings on Low b-value : Validation and Application to Bone Marrow Pathology

  • Byun, Woo-Mok
    • Journal of Yeungnam Medical Science
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    • v.24 no.2
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    • pp.119-128
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    • 2007
  • Purpose : The purpose of this study was a phantom study to measure the diffusion properties of water molecules by steady-state free precession diffusion-weighted imaging (SSFP- DWI) with a low b-value and to determine if this sequence might be useful for application to the evaluation of bone marrow pathology. Materials and methods : 1. The phantom study: A phantom study using two diffusion weighted sequences for the evaluation of the diffusion coefficient was performed. Three water-containing cylinders at different temperatures were designed: phantom A was $3^{\circ}C$, B was $23^{\circ}C$ and C was $63^{\circ}C$. Both SSFP and echo planar imaging (EPI) sequences (b-value: $1000s/mm^2$) were performed for comparison of the diffusion properties. The Signal to noise ratios (SNR) and apparent diffusion coefficient (ADC) values of the three phantoms using each diffusion-weighted sequence were assessed. 2. The Clinical study: SSFP-DWI was performed in 28 patients [sacral insufficiency fractures (10), osteoporotic lumbar compression fractures (10), malignant compression fractures (8)]. To measure the ADC maps, a diffusion-weighted single shot stimulated echo-acquisition mode sequence ($650s/mm^2$) was obtained using the same 1.5-T MR imager Results : For the phantom study, the signal intensity on the SSFP as well as the classic EPI-based DWI was decreased as the temperature increased in phantom A to C. The ADC values of the phantoms on EPI-DWI were $0.13{\times}10^{-3}mm^2/s$ in phantom A, $0.22{\times}10^{-3}mm^2/s$ in B and $0.37{\times}10^{-3}mm^2/s$. in C. The SSFP can be regarded as a DWI sequence in view of the series of signal decreases. Conclusion : Bone marrow pathologies with different diffusion coefficients were evaluated by SSFP-DWI. All benign fractures were hypointense compared to the adjacent normal bone marrow where as the malignant fractures were hyperintense compared to the adjacent normal bone marrow.

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Antihypertensive Effect of Peptide from Sardine Muscle Hydrolyzate on Mild Hypertensive Subjects (정어리 단백질 유래 펩타이드의 경중 고혈압 환자에 대한 강압효과)

  • Kawasaki, T.;Seki, E.;Osajima, K.;Yoshida, M.;Asada, K.;Matsui, T.;Osajima, Y.
    • Proceedings of the Korean Society of Food Hygiene and Safety Conference
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    • 2004.11a
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    • pp.50-58
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    • 2004
  • Valy-Tyrosine (VY) derived from alkaline pretense hydrolyzate of sardine muscles showed the in vitro Angiotensin I converting enzyme (ACE) inhibitory activity and the in vivo antihypertensive effect in SHR. We investigated the antihypertensive effect of the VY on mild hypertensive subjects including subjects with high-normal blood pressure using a randomized double-blind placebo-controlled study. (1) Nineteen subjects (Age 48.9${\pm}$4.3, M/F;18/1) took a 100ml drink either containing 125${\mu}$g of VY or placebo twice daily for 4 weeks. The reductions of the systolic (SBP) and diastolic blood pressure (DBP) were observed in mild hypertensive subjects (n=5) with averages of 17.8${\pm}$2.5 mmHg (p<0.01 vs placebo) and 11.0${\pm}$2.0 mmHg(p<0.05 vs placebo), respectively. Neither SBP nor DBP changed in the subjects of both the placebo group and the high-normal blood pressure group. (2) A randomized double-blind cross-over placebo-controlled study was carried out in 10 mild essential hypertensive subjects (Age 50.6${\pm}$4.6, M/F;10/0). They took a 100ml drink either containing 62.5${\mu}$g of VY or placebo twice daily for 4 weeks alternatively with a 6-week interval. The percent changes in SBP and DBP were -6.9 % and -5.8 % (p<0.05) one week after the VY drink administration, respectively. No adverse effects such as coughing or allergic phenomena could be observed in any of the subjects of drinking VY during the experimental period. These results suggest that the drink containing at least 125${\mu}$g/day of VY may have a significant antihypertensive effect on mild hypertensive subjects without any adverse effects.

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DEVELOPMENT OF HIGH SENSITIVE MODEL OF CARIES ACTIVITY TEST FOR EARLY DIAGNOSIS OF DENTAL CARIES (치아우식증의 조기진단을 위한 고감도 우식활성검사 모형개발)

  • Lee, Sang-Ho;Lee, Chang-Seop
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.1
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    • pp.169-179
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    • 2000
  • The purpose of this study is to develop the system which convert the optical difference of teeth texture between intact enamel and incipient caries lesion into shade difference by laser fluorescence and to develop new and simple caries activity test using laser fluorescence. The experimental design of this study consists of three parts. In first part, a new method for the in vitro assessment of changes in initial enamel caries lesion of Bovine teeth using laser fluorescence is tested. In second part, in vivo assessment undertaken. Number of teeth which showed incipient carious lesion on buccal surface examined by laser fluorescence was compared with the caries activity test of $Cariescreen^{(R)}$ test and other oral environmental test of dDfFtT. In third part, new caries activity test measured by laser fluorescence was developed on the basis of above results and evaluated the sensitivity, specificity, and diagnostic power. Optical density measured by laser fluorescence was increased as increasing the depth of incipient carious lesion and showed high correlation$(\gamma=0.7015)$ with lesion depth. Optical density showed direct proportion to lesion depth. Linear equation was obtained between the optical density and the lesion depth by regression analysis. The result of caries activity test with laser fluorescence showed high correlation with those of $Cariescreen^{(R)}$ test and dDfFtT examination. Caries activity test with laser fluorescence showed 48% of sensitivity, 52% of specificity, and 45% of diagnostic power on the basis of dDfFtT examination, and also showed 48% of sensitivity, 51% of specificity, and 36% of diagnostic power on the basis of $Cariescreen^{(R)}$ test. In regard above result, caries activity test with laser fluorescence considered to be reliable for caries activity test compared with other oral environmental test. and it was also considered to be practical because it would be simple, inexpensive, and time saving method.

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