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Detection of rpoB Gene Mutation in Rifampin-Resistant M. Tuberculosis by Oligonucleotide Chip (Oligonucleotide chip을 이용한 Rifampin 내성 결핵균의 rpoB 유전자 돌연변이 검출)

  • Park, Soon-Kew;Lee, Min-Ki;Chung, Byung-Seon;Kim, Cheol-Min;Chang, Chul-Hun L.;Park, Hee-Kyung;Jang, Hyun-Jung;Park, Seung-Kyu;Song, Sun-Dae
    • Tuberculosis and Respiratory Diseases
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    • v.49 no.5
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    • pp.546-557
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    • 2000
  • Background : Oligonucleotide chip technology has proven to be a very useful tool in the rapid diagnosis of infectious disease. Rifampin resistance is considered as a useful marker of multidrug-resistance in tuberculosis. Mutations in the rpoB gene coding $\beta$ subunit of RNA polymerase represent the main mechanism of rifampin resistance. The purpose of this study was to develop a diagnosis kit using oligonucleotide chip for the rapid and accurate detection of rifampin-resistance in Mycobacterium tuberculosis. Method : The sequence specific probes for mutations in the rpoB gene were designed and spotted onto the glass slide, oligonucleotide chip. 38 clinical isolates of Mycobacterium were tested. A part of rpoB was amplified, labelled, and hybridized on the oligonucleotide chip with probes. Results were analyzed with a laser scanner. Direct sequencing was done to verify the results. Result : The low-density oligonucleotide chip design어 to determine the specific mutations in the rpoB gene of M. tuberculosis accurately detected rifampin resistance associated with mutations in 28 clinical isolates. Mutations at codons 531, 526, and 513 were confirmed by direct sequencing analysis. Conclusion : Mutant detection using oligonucleotide chip technology is a reliable and useful diagnostic tool for the detection of multidrug-resistance in M. tuberculosis.

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AN EXPERIMENTAL STUDY ON THE MICROTENSILE BONDING STRENGTH OF DENTIN TREATED BY $CARISOLV^{TM}$ ($Carisolv^{TM}$ 에 의한 우식제거후 Microtensile Bonding Strength에 관한 연구)

  • Baik, Byeong-Ju;Kwon, Byoung-Woo;Kim, Jae-Gon;Cheon, Cheol-Wan
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.3
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    • pp.389-396
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    • 2002
  • The purpose of this study was to compare the microtensile bonding strength of chemomechanically excavated dentin($Carisolv^{TM}$) to conventional caries removal(bur). The following adhesive systems were used; AB: All-Bond 2(3M, USA), PB: Prime & Bond 2.1(Dentsply, DE), AQ: AQ Bond(sun medical, Japan). 42 human molars with occlusal caries were assigned to 6 groups. Sequential caries removal was controlled with laser fluorescence. Each group was devided as follows; group A, B, C were $Carisolv^{TM}$ applied, group D,E,F were bur used. In group A and D, AB was used as a dentin adhesive. group B,E and group C,F was AQ and AQ was used each. The cavity was filled with composite resin(Z-100). The specimens were sectioned vertically into multiple serial 0.7 mm thick slabs. And then those slabs were sectioned into rectangular parts under 0.7 mm width. Finally 0.7-1.0 mm a right hexahedron shape stick become. Microtensile bonding test was carried out with testing apparatus at cross-head speed of $0.5\;mm/min^{-1}$ and fractured surfaces were observed with scanning electron microscope(JSM-6400, Jeol, Japan). The obtained results were summarized as follows ; 1. In the group of caries removal with $Carisolv^{TM}$, micro-tensile bonding strength decreased to $75.8{\sim}80$ percent of bur used group. 2. In the group of caries removal with $Carisolv^{TM}$, decreased degree of micro-tensile bonding strength is not so different in 3 kinds of dentin adhesives(p<0.05). 3. In the group of caries removal with $Carisolv^{TM}$, microtensile bonding strength of AB, PB, AQ was 32.6MPa(2.4), 30.1Mpa (1.8), 21.2Mpa(1.9). 4. In the group of caries removal with Bur and $Carisolv^{TM}$, microtensile bonding strength of AQ was significantly lower than that of AB and PB(p<0.01).

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IN VITRO COMPARISON OF VARIOUS DIAGNOSTIC METHODS OF OCCLUSAL CAR10US LESIONS (교합면 우식병소의 다양한 진단법에 관한 비교연구)

  • Kim, Jae-Gon;Kim, Young-Jin;Kim, Young-Sin;Baik, Byeong-Ju
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.4
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    • pp.613-619
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    • 2001
  • The aims of this study were to compare the accuracy, sensitivity and specificity of cnventional visual examination, radiography and a new laser fluorescence method, KaVo Diagnodent, for the detection of occlusal caries lesions. One hundred sound human premolars and molars which had no restorations or interproximal cavities were tested by three methods. Tooth lesions depth was assessed at histologic examination using Caries detector dye The following results were obtained. 1. Diagnodent show 7.8 in sound tooth, 25.4 in initial caries, 30.5 in enamel caries, and 53.8 in dentin caries with average score 2. Spearman and Pearson relation coefficient was high between tooth-specimen test with dye and Diagnodent(0.736, 0.619), visual examination(0.664, 0.666), and was low between tooth-specimen test with dye and radiographic examination(P<0.01, total) 3. Accuracy of occlusal caries was highest on Diagnodent(65%) and lowest on radiographic examination(35%) 4. In initial caries, the sensitivity and specificity of Diagnodent method was the highest. In enamel caries, the sensitivity of visual examination was the highest and specificity of Diagnodent method was the highest. In dentinal caries, the sensitivity and specificity of Diagnodent method was the highest and sensitivity of visual examination was the lowest.

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Elemental analysis of the fluoride varnish effects on root caries initiation (불소 바니쉬 도포 후 초기 치근 우식 발현에 대한 정량원소분석)

  • Park, Se-Eun;Yi, Kee-Wook;Kim, Hae-Young;Son, Ho-Hyun;Chang, Ju-Hea
    • Restorative Dentistry and Endodontics
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    • v.36 no.4
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    • pp.290-299
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    • 2011
  • Objectives: The usage of fluoride varnish for a moderate to low caries-risk group has not been well validated. This study aimed to evaluate the preventive and therapeutic efficacies of fluoride varnish on the initiated root caries. Materials and Methods: Ten premolars were sectioned into quarters, further divided into two windows, one of which was painted with Fluor Protector (1,000 ppm fluoride, Ivoclar Vivadent). An initial lesion with a well-preserved surface layer was produced by pH cycling. Scanned line analysis using energy dispersive spectrometry determined the weight percentages of Ca and P in the demineralized layer. Scanning Electron microscopy and confocal laser scanning microscopy (CLSM) evaluated the varnish-applied root surfaces. Results: The mean lesion depth (SD) was 12.3 (2.6) ${\mu}m$ (single cycling) and 19.6 (3.8) ${\mu}m$ (double cycling). Double cycling extended the lesion depth, but induced no more mineral loss than single cycling (p < 0.05). The mean weight percentages of Ca and P between groups with and without varnish were not significantly different (p < 0.05). A CLSM showed varnish remained within 15 ${\mu}m$ of the surface layer. Conclusions: When a mild acid challenge initiated root tissue demineralization, the application of low-concentration fluoride varnish did not influence the lesion depth or the mineral composition of the subsurface lesion.

Physiological and Proteome Responses of Korean F1 maize (Zea mays L.) Hybrids to Water-deficit Stress during Tassel Initiation (옥수수 영양생장기 한발 스트레스에 의한 광합성의 생리적 반응 및 프로테옴 변화 분석)

  • Bae, Hwan Hee;Kwon, Young-Sang;Son, Beom-Young;Kim, Jung-Tae;Go, Young Sam;Kim, Sun-Lim;Baek, Seong-Bum;Shin, Seonghyu;Kim, Sang Gon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.64 no.4
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    • pp.422-431
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    • 2019
  • Severe droughts in spring have occurred frequently in Korea in recent years, exerting a critical impact on corn yield. Therefore, it is necessary to find physiological and/or molecular indicators of the response to drought stress in maize plants. In this study, we investigated the effects of water-deficit stress on two Korean elite F1 maize hybrids, Ilmichal and Gwangpyeongok, by withholding water for 10 days at tassel initiation. The water deficit drastically reduced the relative leaf water content, leaf number, leaf area, and stem length, leading to dry matter reduction. Moreover, it reduced the SPAD values and stomatal conductance of leaves in drought-stressed plants of both hybrids. Importantly, the number of leaves and SPAD value were non-destructive and easy to investigate in response to water-deficit stress, suggesting that they may be useful indicators for screening drought-tolerant genetic resources. We detected more than 100 spots that were differentially accumulated under drought stress. Of these spots, a total of 21 protein spots (≥1.5-fold) from drought-exposed maize leaves were successfully analyzed by MALDI-TOF-TOF mass spectrometry. Functional annotation using Gene Ontology analysis revealed that most of the identified proteins were involved in carbohydrate metabolism, stress response fatty acid catabolism, photosynthesis, energy metabolism, and transport. The protein expression levels were increased in both Ilmichal and Gwangpyeongok, except for triosephosphate isomerase, fructose-bisphosphate aldolase, and an uncharacterized protein. The lactoylglutathione lyase delta (3,5)-delta (2,4)-dienoyl-CoA isomerase was overexpressed in Gwangpyeongok only. The results obtained from this study suggest that the drought-specific genes may be useful as molecular markers for screening drought-tolerant maize genotypes.

Effect of Fusion Condition on In Vitro Development of Caprine Cloned Oocytes with Nuclear Transfer (재래산양에 있어서 핵이식란의 융합조건이 융합 및 체외발달에 미치는 영향)

  • 박희성;김태숙;이윤희;정수영;이명열;홍승표;박준규;김충희;정장용
    • Reproductive and Developmental Biology
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    • v.28 no.2
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    • pp.127-132
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    • 2004
  • This study was conducted to examine the effects of electric stimulation conditions on in vitro developmental ability of caprine embryos after somatic cell nuclear transfer. Recipient oocytes were surgically collected after superovulation by using CIDR and FSH, PMSG, hCG and estrous synchronization in Korean native goats. The caprine ear cells were cultured in vitro in serum-starvation condition (TCM-l99 + 0.5% FBS) for 3 to 5 days of cell confluence. The zona pellucida of in vivo and in vitro matured oocytes were partially drilled using laser system. Single somatic cell was individually transferred into the enucleated oocyte. The reconstructed oocytes were electrically fused with 0.3M mannitol. After the electofusion, embryos were activated by electric stimulation or Ionomycin + 6-DMAP. Nuclear transfer embryos were cultured in mSOF medium supplemented with 0.8% BSA 6∼7 days at 39 , 5% $CO_2$, 5% $O_2$, 90% $N_2$. The fusion rate of donor cells was 60.4% and 40.3 % in ear cell and fetal fibroblast, and cleavage rate were 40.6% and 48.2%, respectively. No significant difference was found in the fusion and cleavage rate in different donor cells. Nuclear transferred oocytes were fused by electric pulses of 1.30∼1.40, 2.30∼2.39 and 2.40∼2.46 ㎸/cm. There was no significant difference among different electric pulses in fusion rates (26.7, 34.8 and 43.8%). The cleavage rate was higher (p<0.05) in 1.30∼1.40 ㎸/cm (82.9%) than 2.30∼2.39 ㎸/cm (43.8%) and 2.40∼2.46 ㎸/cm. (51.8%). The fusion rates of recipient oocyte source were 1st (43.5% and 23.6%), 2nd (55.7% and 39.2%) and 3rd (66.1% and 52.8%) in in vivo and in vitro oocytes. However, fusion ratee were significantly higher (p<0.05) in in vivo than in vitro oocyte. The cleavage rate of fused oocytes from in vivo and in vitro sources were 52.6% and 54.4%, respectively. No significant difference was found in the cleavage rate according to the recipient oocyte source. These results suggest that factors such as field pulse of electric stimulation and oocyte source could affect in vitro developmental ability of nuclear transplanted caprine oocytes.

A Study on the Cosmeceutical activities of Prunus Sargentii R. (산벚나무(Prunus sargentii R.) 수피의 화장품활성에 관한 연구)

  • Park, Jung-Mi;Lee, Jin-Young;Park, Tae-Soon;Hyun, Sok-Jun;Kim, Han-Hyuk;Cho, Young-Je;Kwon, O-Jun;Son, Ae-Ryang;Kim, Dong-Seok;An, Bong-Jeun
    • Applied Biological Chemistry
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    • v.51 no.1
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    • pp.70-78
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    • 2008
  • Prunus sargentii R. of Rosaceae familiy, has been reported to have radical scavenging activity and anti-inflammatory effect. On these facts, biological activity and safety test were conducted to evaluate biological activities of the extracts of P. sargentii R. as a potential pharmaceutical ingredient. The electron donating ability of its ethanol extracts at a 500 ppm level showed 92%, which was higher than that of hot water extract (59%), the superoxide dismutase (SOD)-like activity of the water extract of P. sargentii R. was about 50%, the ethanol extract of P. sargentii R. was about 40% at 1,000 ppm concentration. Xanthine oxidase inhibition by the water extract of P. sargentii R. was about 40% and that by the ethanol extract was 60% respectively at 500 ppm concentration. From the measurement on lipid oxidation, the $Cu^{2+}$ chelating effect of the ethanol extract was higher than that of hot water extract. The $Fe^{2+}$ chelating effect was also shown to be about 80% at a 500 ppm concentration in both hot water extract and ethanol extract. The tyrosinase inhibition effect related to skin-whitening was 26% by hot water extract and 20% by ethanol extract respectively at a 1,000 ppm. Hyaluronidase inhibition activity related to the anti-inflammation effect was 96% in ethanolic extract at a 500 ppm. Clear zones formed by P. sargentii R. against the human skin-resident micro-flora such as Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Propionibacterium acnes indicated that antimicrobial activity of the ethanol extract was higher than that of the hot water extract.

Photodynamic Therapy with Photofrin Reduces Invasiveness of U87 Malignant Human Glioma Cells (교모세포종 세포주 U87에서 Photofrin을 사용한 광역학 치료가 종양 침습성에 미치는 영향)

  • Woo, Hye Kyung;Cho, Kyung-Keun;Rha, Hyung Kyun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Park, Hea Kwan;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup2
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    • pp.189-196
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    • 2001
  • Objective : We tested the hypothesis that photodynamic therapy(PDT) with Photofrin inhibits tumor invasion of U87 human glioma cells using several in vitro assay to measure tumor invasiveness. The effects of PDT on cell growth, directional migration and cell invasion were investigated. Material and Method : Tumor cells were treated with Photofrin at various doses and at a fixed optical(632nm) dose of $100mJ/cm^2$. Cytotoxicity was tested using the MTT method. Invasion assays including the matrigelartificial basement membrane barrier migration and spheroid confrontation with confocal microscopic analysis were used to study the relationship between PDT and invasiveness. Result : U87 cells showed a dose dependent cytotoxic response to increasing Photofrin dose. Data from the matrigel artificial basement membrane assay indicate that PDT inhibits the U87 cell migration dose dependently. Low doses of subcytotoxic PDT treatment, such as 2.5ug/ml Photofrin dose, also appeared to significantly inhibit migration of U87 cells(p<0.05). In co-cultures between U87 cell spheroids and brain aggregates, progressive invasion with destruction of the brain aggregate occurs. The extent of tumor cell infiltration and proportion or intact brain aggregate remaining after 24h differs in Photofrin PDT treated versus Photofrin only control, with changes suggestive of a dose-response effect. Conclusion : our data indicate that PDT with Photofrin significantly inhibits the invasiveness of U87 cells, and this inhibition is dose dependent.

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Effects of acid-treatment conditions on the surface properties of the RBM treated titanium implants (산-처리 조건이 RBM처리한 티타늄 임플란트의 표면 특성에 주는 영향)

  • Lee, Han-Ah;Seok, Soohwang;Lee, Sang-Hyeok;Lim, Bum-Soon
    • Korean Journal of Dental Materials
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    • v.45 no.4
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    • pp.257-274
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    • 2018
  • The purpose of this study was to evaluate the effect of acid-treatment conditions on the surface properties of the RBM (Resorbable Blast Media) treated titanium. Disk typed cp-titanium specimens were prepared and RBM treatments was performed with calcium phosphate ceramic powder. Acid solution was mixed using HCl, $H_2SO_4$ and deionized water with 4 different volume fraction. The RBM treated titanium was acid treated with different acid solutions at 3 different temperatures and for 3 different periods. After acid-treatments, samples were cleaned with 1 % Solujet solution for 30 min and deionized water for 30 min using ultrasonic cleanser, then dried in the electrical oven ($37^{\circ}C$). Weight of samples before and after acid-treatment were measured using electric balance. Surface roughness was estimated using a confocal laser scanning microscopy, crystal phase in the surface of sample was analyzed using X-ray diffractometer. Surface morphology and components were evaluated using Scanning Electron Microscope (SEM) with Energy Dispersive X-ray spectroscopy (EDX) and X-ray Photoemission Spectroscopy (XPS). Values of the weight changes and surface roughness were statistically analyzed using Tukey-multiple comparison test (p=0.05). Weight change after acid treatments were significantly increased with increasing the concentration of $H_2SO_4$ and temperature of acid-solution. Acid-treatment conditions (concentration of $H_2SO_4$, temperature and time) did not produce consistent effects on the surface roughness, it showed the scattered results. From XRD analysis, formation of titanium hydrides in the titanium surface were observed in all specimens treated with acid-solutions. From XPS analysis, thin titanium oxide layer in the acid-treated specimens could be evaluated. Acid solution with $90^{\circ}C$ showed the strong effect on the titanium surface, it should be treated with caution to avoid the over-etching process.

Effects on the Thermal Change of the Face Follow Electroacupunctyre on Hapkok($LI_4$), Sangan($LI_3$) (合谷($LI_4$), 三間($LI_3$)의 電針刺戟이 顔面部 領域 溫度變化에 미치는 影響)

  • Yun, Jeong-hun;Kim, Jong-Han;Hwang, Chung-yeon;Lim, Kyu-sang
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.12 no.2
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    • pp.222-247
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    • 1999
  • The back ground and purpose : The acupuncture of oriental medicine is very important in treatments. Until now it has been researched according to the meridian and qi xue(氣血) phenomenon of oriental medicine's theory. As electroacupuncture is one of acupuncture treatments, it will show more objective index to observe the meridian and qi xue(氣血) phenomenon. And then, I studied the effects on the thermal change of the face following electroacupuncture treatment. Objective and Methods : This study was performed from January 1999 to March 1999 on 10 healthy students. The objective was divided into three groups, those were the control group A(n=10), the group B(n=10) of electroacupuncture on Hapkok($LI_4$), Samgan($LI_3$) and the group C(n=10) of electroacupuncture on Shinmun($H_7$), T' ongni($H_5$). First, in the control group A, we took a picture for 10 men without any stimulation with the Digital Infrared Thermograph Imaging(D.I.T.I.) and did 3min after, 10min after, 15min after, 25min after, 45min after respectively. Second, in the electroacupuncture treatment group B, we took a picture for 10 men without any stimulation, and then treat electroacupuncture on Hapkok($LI_4$), Samgan($LI_3$) and took a picture immediately(3min after), 10min after, 15min after and remove needle and took a picture in the same way respectively. Third, in the electroacupuncture treatment group C, we took a picture for 10 men without any stimulation, and then treat electroacupuncture on Shinmun($H_7$), T'ongni($H_5$) and took a picture in the second way respectively. Results: 1. In healthy men, average skin temperture about Yonghyang($LI_{20}$) area was higher than Soryo($G_{25}$) or Chich'ang($S_4$) area. They were Soryo($G_{25}$) area $31.495{\pm}0.766^{\circ}C$, Rt. Yonghyang($LI_{20}$) area $31.664{\pm}0.936^{\circ}C$, Lt. Yonghyang ($LI_{20}$)area $31.686{\pm}0.767^{\circ}C$, Rt. Chich'ang($S_4$) area $31.226{\pm}0.875^{\circ}$, Lt. Chich'ang ($S_4$) area $31.453{\pm}0.855^{\circ}C$. 2. In the control group A, the skin temperature of Soryo($G_{25}$) showed the increase or decrease in below ${\Delta}0.1^{\circ}C\;except\;0.265{\pm}0.594^{\circ}C$ in 25min, but not significantly. 3. About Soryo($G_{25}$) area, the skin temperature decreased significantly after electroacupuncture immediately. ${\Delta}T $of the group B was $-0.970{\pm}0.87\;1^{\circ}C$, which was larger than one of the group C which was $-0.707{\pm}0.624^{\circ}C$ at 3min. And then ${\Delta}T$ of the group C was increase valuable at 25min, 45min. 4. About Yonghyang($L1_{25}$) area, the left ${\Delta}T$ of the group B showed below $0.2^{\circ}C$ or so in contrast to the right it. In the group C, on the both side showed continous increase of temperature as following times. 5. About Chich'ang($S_4$) area, the skin temperature increased valuable $0.3^{\circ}C$ or so on the both side and later inclined to decrease in the group B but not significantly. In the group C, it increased valuable on the both side. 6. The skin temperature of electroacupuncture treatment group B, C were more increase than the control group A except Lt. Yonghyang($LI_{20}$) area in the group B. The temperature of group C were more increase than the group B wholly. Conclusion : The above results indicate that D.I.T.I. is a useful method to observe and fallow-up the effects and the changes by electroacupuncture stimulation on objective evaluation of phenomenon for the meridian system and character. Thus, continuous thermographic study will be needed for more clinical application such as acupuncture and medicine or laser therapy according to oriental medicine.

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