• Journal of Acupuncture Research
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• v.23 no.6
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• pp.117-132
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• 2006

Objectives : The purpose of this study is to investigate the effect of Ulmus davidiana Planch herbal acupuncture solution in LPS-stimulated RAW 264.7 cells and mouse knee joints, perfom1ed several experimental items: those are MIF mRNA, MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS mRNA, iNOS, NO, synovial hyperplasia, angiogenesis and fibrosis. Methods : In order to observe mRAN expression of MIF and iNOS in LPS-stimulated RAW 264.7 cells, RT-PCR was used. NO production in LPS-stimulated RAW 264.7 cells was measured by nitrite assay. All the female BALB/c mice were bred and maintained in pathogen-free mouse colonies and were 6 weeks of age on beginning of the experiment. The experimental model of RA was induced by injection of $50{\mu}g/kg$ LPS. Ulmus davidiana Planch herbal acupuncture solution was injected into either S 35 (犢鼻) or EX-LE 202 (內膝眼) of mice in turn daily for 19 days. Immunohistochemical staining was carried out to assess $TNF-{\alpha}$, $NF-{\kappa}B$ p65 and iNOS expression in synovial membrane. Synovial hyperplasia, angiogenesis and fibrosis in synovial membrane was observed with a microscope. Results : 1. Ulmus davidiana Planch herbal acupuncture solution inhibited mRNA expression of MIF and iNOS in dependence on a density of it in LPS-stimulated RAW 264.7 cells. 2. Ulmus davidiana Planch herbal acupuncture solution decreased synovial hyperplasia, angiogenesis and fibrosis in LPS-stimulated mouse knee joints. 3. Ulmus davidiana Planch herbal acupuncture solution curtailed production of MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS in LPS-stimulated mouse knee joints. Conclusion : On the basis of these results, It was shown that Ulmus davidiana Planch herbal acupuncture solution is significantly able to inhibit the production of MIF as a top in cytokines related to inflammatory or irrlll1une responses. Our results may provide that Ulmus davidiana Planch herbal acupuncture solution has beneficial effect in not only RA but other inflammatory or immune deases.

• The Journal of Internal Korean Medicine
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• v.34 no.2
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• pp.178-191
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• 2013

Objectives : In this study, we made an effort to investigate the protective mechanism of Ukgan-san (UGS) extracts on hypoxia-induced C6 glial cell death. Methods : The cell viability was assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MMT) assay and cell morphological changes were analysed with microscope after staining with crystal violet (CV). Reactive oxygen species (ROS) formation was assessed by flow cytometer after staining with 2'7'-dichlorofluorescein diacetate (DCF-DA). We also analyzed expression of hypoxia-inducible factor-1 alpha (HIF-$1{\alpha}$) and p53, processing of procaspase-3 and procyclic acidic repetitive protein (PARP) by western blot method. Results : We estimated the elevated cell viability by UGS extract on $CoCl_2$-induced C6 glial cells. UGS attenuated $CoCl_2$-induced ROS formation in C6 glial cells and also showed a protective activity compared to antioxidants and exhibited abrogation of LDH-released by $CoCl_2$. UGS suppressed the typical apoptotic cell death markers, caspase-3 and PARP activation. UGS inhibited $CoCl_2$-induced HIF-1${\alpha}$ expression which is known as a major regulator for hypoxia-induced cell death, and suppressed p53 expression. Conclusions : These results suggest that UGS extract contains protective constituents for hypoxia-induced C6 glial cell death.

• Journal of Embryo Transfer
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• v.32 no.4
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• pp.311-317
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• 2017

Cryopreservation of oocytes by vitrification technique may contribute a lot in the field of reproductive biotechnology. The objectives of the present study were to evaluate the effectiveness of two cryo-devices for vitrification of immature oocytes of indigenous zebu cows. Slaughter house derived immature cumulus-oocyte-complexes (COCs) of cows were vitrified using 15% dimethyl sulphoxide (DMSO) as cryoprotective agent (CPA) with 0.5 mol sucrose in TCM 199 supplemented with 20% FBS. Vitrification of COCs was completed after immediate plunging of COCs loaded cryotop or French mini straw into the liquid nitrogen ($LN_2$). Then the COCs containing cryotop or French mini straws were warmed in 0.25 mol sucrose and 20% FBS supplemented TCM 199 followed by in vitro culture in $50{\mu}l$ droplets of bicarbonate buffered TCM 199 supplemented with 10% FBS, pyruvate, FSH and oestradiol for 24 hrs at $39^{\circ}C$ with 5% CO2 in humidified air. After maturation culture, oocytes were denuded and examined under inverted microscope for presence of polar body as the indication of maturation. Denuded oocytes were also stained by whole mount technique using 1% orcein to examine the maturation by presence of MII chromosomes. The in vitro maturation rate was significantly (p<0.05) higher in oocytes vitrified and warmed using crytop ($47.1{\pm}6.9%$) than that of French mini straw ($15.9{\pm}12.5%$). Moreover, in vitro maturation rate was significantly (p<0.05) highe r in control oocytes (not vitrified) ($84.5{\pm}14.2%$) than that of vitrified oocytes. In conclusion, cryotop is better than French mini straw as cryo-device for vitrification of bovine immature oocytes.

• Restorative Dentistry and Endodontics
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• v.41 no.1
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• pp.12-21
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• 2016

Objectives: To evaluate the effects of three acids on the microhardness of set mineral trioxide aggregate (MTA) and root dentin, and cytotoxicity on murine macrophage. Materials and Methods: OrthoMTA (BioMTA) was mixed and packed into the human root dentin blocks of 1.5 mm diameter and 5 mm height. Four groups, each of ten roots, were exposed to 10% citric acid (CA), 5% glycolic acid (GA), 17% ethylenediaminetetraacetic acid (EDTA), and saline for five minutes after setting of the OrthoMTA. Vickers surface microhardness of set MTA and dentin was measured before and after exposure to solutions, and compared between groups using one-way ANOVA with Tukey test. The microhardness value of each group was analyzed using student t test. Acid-treated OrthoMTA and dentin was examined by scanning electron microscope (SEM). Cell viability of tested solutions was assessed using WST-8 assay and murine macrophage. Results: Three test solutions reduced microhardness of dentin. 17% EDTA demonstrated severe dentinal erosion, significantly reduced the dentinal microhardness compared to 10% CA (p = 0.034) or 5% GA (p = 0.006). 10% CA or 5% GA significantly reduced the surface microhardness of set MTA compared to 17% EDTA and saline (p < 0.001). Acid-treated OrthoMTA demonstrated microporous structure with destruction of globular crystal. EDTA exhibited significantly more cellular toxicity than the other acidic solutions at diluted concentrations (0.2, 0.5, 1.0%). Conclusions: Tested acidic solutions reduced microhardness of root dentin. Five minute's application of 10% CA and 5% GA significantly reduced the microhardness of set OrthoMTA with lower cellular cytotoxicity compared to 17% EDTA.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.1
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• pp.1-17
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• 2020

Three-dimensional microscopic approaches in histopathology display multiplex properties that present puzzling questions for specimens as related to their comprehensive volumetric information. This information includes spatial distribution of molecules, three-dimensional co-localization, structural formation and whole data set that cannot be determined by two-dimensional section slides due to the inevitable loss of spatial information. Advancement of optical instruments such as two-photon microscopy and high performance objectives with motorized correction collars have narrowed the gap between optical theories and the actual reality of deep tissue imaging. However, the benefits gained by a prolonged working distance, two-photon laser and optimized beam alignment are inevitably diminished because of the light scattering phenomenon that is deeply related to the refractive index mismatch between each cellular component and the surrounding medium. From the first approaches with simple crude refractive index matching techniques to the recent cutting-edge integrated tissue clearing methods, an achievement of transparency without morphological denaturation and eradication of natural and fixation-induced nonspecific autofluorescence out of real signal are key factors to determine the perfection of tissue clearing and the immunofluorescent staining for high contrast images. When performing integrated laboratory workflow of tissue for processing frozen and formalin-fixed tissues, clear lipid-exchanged acrylamide-hybridized rigid imaging/immunostaining/in situ hybridization-compatible tissue hydrogel (CLARITY), an equipment-based tissue clearing method, is compatible with routine procedures in a histopathology laboratory.

• Journal of Acupuncture Research
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• v.24 no.2
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• pp.211-220
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• 2007

Objectives : This study was to investigate the effect of high frequency electroacupuncture at $ST_{36}$ acupuncture point on the Complete Freund's Adjuvant (CFA) induced rat arthritis pain model. Methods : Arthritis was induced by intradermal injection of CFA into base of tail. Experimental groups were divided into 4 groups; Normal, Control and Jok-Samri ($ST_{36}$) and Non-Acupuncture point (NA). Normal group, non-arthritic group, was injected with normal saline，and the others groups were injected CFA. $ST_{36}$ group was treated by 120 Hz electroacupuncture at $ST_{36}$ acupuncture point, and NA group was treated by 120 Hz electroacupuncture at non-acupuncture point. Each groups were evaluated by the change of c-fos positive neurons in periaqueductal gray (PAG) and hippocampus by using an image analyzer and a microscope. Results: - In the PAG region, the number of fos-positive cells in the $ST_{36}$ group ($42.37{\pm}5.08$) were significantly (p<0.05) decreased compared with the control group ($64.56{\pm}6.35$). - In the PAG region, the number of fos-positive cells in the NA group were meaninglessly decreased compared with the control group - In the Cornu Ammonis(CA)l region of hippocampus, the number of fos-positive cells in the $ST_{36}$ group ($7.00{\pm}1.08$) and NA group ($5.56{\pm}2.01$) were significantly (p<0.05) decreased compared with the control group ($13.81{\pm}1.24$). - In the dentate gyrus region of hippocampus, the number of fos- positive cells in the $ST_{36}$ group ($10.75{\pm}0.98$) and NA group ($6.56{\pm}0.78$) were significantly (p$26.45{\pm}1.82$). Conclusions : It is expected that high frequency electroacupuncture can be used a treatment of arthritic pain.

• Journal of Acupuncture Research
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• v.25 no.4
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• pp.1-9
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• 2008

Objectives : This study was to investigate the effect of low frequency electroacupuncture according to acupoints combination on NOS positive neurons in the PAG and Hippocampus of Rat with adjuvant induced Rheumatoid arthritis. Methods : Arthritis was induced by intradermal injection of FCA into base of tail. Experimental group were divided into 6 group ; Normal, Control, $ST_{36}$, $SP_9$, $ST_{36}+SP_9$, and NA. Normal group was injected with normal saline and not treated electroacupuncture. Control group was injected with FCA and not treated electroacupuncture. ST36 group was injected with FCA and treated by 2 Hz electroacupuncture at $ST_{36}$. $SP_9$ group was injected with FCA and treated by 2 Hz electroacupuncture at $SP_9$. $ST_{36}+SP_9$ group was injected with FCA and treated by 2 Hz electroacupuncture at $ST_{36}+SP_9$. NA group was injected with FCA and treated by 2 Hz electroacupuncture at non acupoint. Each groups were evaluated by the number of NOS-positive neurons in PAG and hippocampus by using an image analyzer and a microscope. Results : 1. The number of NOS-positive neurons of the NA group were significantly decreased in the CA2-3. 2. The number of NOS-positive neurons of the ST36 group were significantly decreased in the PAG and CA2-3. 3. The number of NOS-positive neurons of the SP9 group were significantly decreased in the PAG and DG 4. The number of NOS-positive neurons of the ST36+SP9 group were significantly decreased in the PAG, CA1 and DG. Conclusions : It is expected that low frequency electroacupuncture can be used a treatment of rheumatoid arthritic pain and that treatment of electroacupuncture at combined acupoints is more efficient than treatment of low frequency electroacupuncture at one acupoint.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.201-206
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• 2014

The objectives of the study were to determine an effective culture dish, culture duration and protein supplementation in medium for in vitro maturation (IVM) of oocytes of native zebu cows in Bangladesh. The ovaries of cows were collected from local slaughterhouse followed by aspiration of follicular fluid. The cumulus-oocyte-complexes (COCs) with more than 3 compact cumulus cell layers were cultured in tissue culture medium (TCM) 199 for maturation. The maturation of oocytes was determined by observing polar body under microscope. To determine an effective culture dish, 130 COCs derived from 48 ovaries in a well of 4-well dish and 102 COCs derived from 36 ovaries in drops covered with mineral oil within 35 mm petri dish were cultured for 24 hours. The rate of maturation of oocytes did not vary between 4-well dish ($51.3{\pm}15.0%$) and drops in petri dish ($52.4{\pm}11.6%$). To determine the effective culture duration, 185 COCs derived from 62 ovaries were cultured in drops for 18, 21, 24 and 27 hours. The rate of maturation of occytes ranged from $51.9{\pm}9.4%$ (18 hours) to $59.0{\pm}17.1%$ (27 hours) and the difference in maturation rate among different culture durations was not significant (P>0.05). To determine an effective protein supplementation, 63 oocytes from 19 ovaries were cultured separately in TCM 199 supplemented with either fetal bovine serum (FBS) or bovine serum albumin (BSA). The rate of maturation was significantly (P<0.01) higher in medium supplemented with FBS ($55.63{\pm}16.19%$) than that of BSA ($14.82{\pm}9.36%$). In conclusion, COCs of native zebu cows can be cultured for IVM either in 4-well culture dish or droplets in petri dish for 18 to 27 hours in medium supplemented with FBS.

• Journal of Korean Society of Occupational and Environmental Hygiene
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• v.21 no.2
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• pp.110-115
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• 2011

Objectives: The aim of this study is to survey for finding asbestos containing equipment at the laboratories using picture based questionnaire and polarized light microscopic analysis. Methods: This study was conducted from 2009 to 2010 at a university in Seoul. In 2009, picture based questionnaire was distributed to 100 laboratories during the regular laboratory air quality monitoring. In 2010, we emailed all professors of the same university who have laboratories to participate voluntarily this survey. For the laboratories consented to participate survey, picture based questionnaire was distributed and collected. Suspected asbestos containing material and apparatus were collected at the laboratories which replied they have suspected material and equipment. Collected samples were analyzed with polarized light microscope at the laboratory accredited by ministry of employment and labor in Korea. Results: Total of 18 out of 100 laboratories reported that they had suspected asbestos containing equipment in 2009. Twenty-three samples were collected and three samples (13%), one heating mantle and two pairs of insulation gloves, contained asbestos. Thirty four laboratories reported they had suspected asbestos containing material or equipment in 2010. Sixty samples were collected and four of them (6%), two pairs of insulation gloves, one packing rope in dry oven and, one pair of tongs, contained asbestos. All founded asbestos was chrysotile and the content of chrysotile was more than 90% for all equipment except heating mantle which has less than 1%. Conclusions: We confirmed that asbestos was still used at the laboratories though strict regulations on asbestos use in Korea. The method of picture based questionnaire invented in this study could be applied for asbestos survey to other research institute or university where there are many laboratories because of its simplicity and accessibility without huge man power, cost and time.

• Restorative Dentistry and Endodontics
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• v.35 no.6
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• pp.486-491
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• 2010

Objectives: This study examined the effect of 2% chlorhexidine on the ${\mu}TBS$ of a direct composite restoration using one-step self-etch adhesives on human dentin. Materials and Methods: Twenty-four extracted permanent molars were used. The teeth were assigned randomly to six groups (n = 10), according to the adhesive system and application of chlorhexidine. With or without the application of chlorhexidine, each adhesive system was applied to the dentin surface. After the bonding procedure, light-cure composite resin buildups were produced. The restored teeth were stored in distilled water at room temperature for 24 hours, and then cut and glued to the jig of the microtensile testing machine. A tensile load was applied until the specimen failed. The failure mode was examined using an operating microscope. The data was analyzed statistically using one-way ANOVA, Student's t-test (p < 0.05) and Scheffet's test. Results: Regardless of the application of chlorhexidine, the Clearfil $S^3$ Bond showed the highest ${\mu}TBS$, followed by G-Bond and Xeno V. Adhesive failure was the main failure mode of the dentin bonding agents tested with some samples showing cohesive failure. Conclusions: The application of 2% chlorhexidine did not affect the ${\mu}TBS$ of the resin composite to the dentin using a one-step self-etch adhesive.