• Journal of Korean Biological Nursing Science
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• 제15권4호
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• pp.257-263
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• 2013

Purpose: The purpose of this study was to investigate the effect of Ulmus root-bark dressing in fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) of induced pressure ulcers in rats. Methods: 54 male Sprague-Dawley rats were used and randomly divided into 2 groups. The rats were anesthetized and pressure ulcers were induced at 140 mmHg for three hours, using a personally-designed pressing apparatus. Ulmus dressing was applied in the experimental group (n=27) and saline gauze dressing in the control group (n=27). Each of the dressings was changed every other day, and after a month, the wounds were examined by microscopy biweekly for 20 weeks. Results: After 4 weeks, the epidermis of the wounds was recovered, but inflammatory infiltration of the dermis was remained. After 6 weeks, inflammatory cells were diminished and the number of capillaries was decreased. Examined by immunofluorescence staining, the FGF positive cells in the experimental group changed negatively after 18 weeks, but the control group still existed even after 20 weeks. VEGF positive cells in the experimental group also changed negatively after 20 weeks, but the control group still existed. Conclusion: The findings of this study demonstrate that Ulmus dressing is effective in minimizing scar formation and inflammatory reaction by decreasing FGF and VEGF in the terminal phase of wound healing.

• Molecular & Cellular Toxicology
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• 제5권3호
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• pp.230-236
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• 2009

Acrolein, a known toxin in cigarette smoke, is the most abundant electrophilic $\alpha$, $\beta$-unsaturated aldehyde to which humans are exposed in a variety of environmental pollutants, and is also product of lipid peroxidation. Increased unsaturated aldehyde levels and reduced antioxidant status plays a major role in the pathogenesis of various diseases such as diabetes, Alzheimer's and atherosclerosis. The findings reported here show that low concentrations of acrolein induce heme oxygenase-1 (HO-1) expression in RAW 264.7 macrophages. HO-1 induction by acrolein and signal pathways was measured using reverse transcription-polymerase chain reaction, Western blot and immunofluorescence staining analyses. Inhibition of extracellular signal-regulated kinase activity significantly attenuated the induction of HO-1 protein by acrolein, while suppression of Jun N-terminal kinase and p38 activity did not affect induction of HO-1 expression. Moreover, rottlerin, an inhibitor of protein kinase $\delta$, suppressed the upregulation of HO-1 protein production, possibly involving the interaction of NF-E2-related factor 2 (Nrf2), which has a key role as a HO-1 transcription factor. Acrolein elevated the nuclear translocation of Nrf2 in nuclear extraction. The results suggest that RAW 264.7 may protect against acrolein-mediated cellular damage via the upregulation of HO-1, which is an adaptive response to oxidative stress.

• 대한약침학회지
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• 제17권1호
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• pp.59-69
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• 2014

Objectives: In homeopathy, it is claimed that more homeopathically-diluted potencies render more protective/curative effects against any disease condition. Potentized forms of Condurango are used successfully to treat digestive problems, as well as esophageal and stomach cancers. However, the comparative efficacies of Condurango 6C and 30C, one diluted below and one above Avogadro's limit (lacking original drug molecule), respectively, have not been critically analyzed for their cell-killing (apoptosis) efficacy against lung cancer cells in vitro, and signalling cascades have not been studied. Hence, the present study was undertaken. Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays were conducted on H460-non-small-cell lung cancer (NSCLC) cells by using a succussed ethyl alcohol vehicle (placebo) as a control. Studies on cellular morphology, cell cycle regulation, generation of reactive oxygen species (ROS), changes in mitochondrial membrane potential (MMP), and DNA-damage were made, and expressions of related signaling markers were studied. The observations were done in a "blinded" manner. Results: Both Condurango 6C and 30C induced apoptosis via cell cycle arrest at subG0/G1 and altered expressions of certain apoptotic markers significantly in H460 cells. The drugs induced oxidative stress through ROS elevation and MMP depolarization at 18-24 hours. These events presumably activated a caspase-3-mediated signalling cascade, as evidenced by reverse transcriptase-polymerase chain reaction (RT-PCR), western blot and immunofluorescence studies at a late phase (48 hours) in which cells were pushed towards apoptosis. Conclusion: Condurango 30C had greater apoptotic effect than Condurango 6C as claimed in the homeopathic doctrine.

• Applied Microscopy
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• 제47권2호
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• pp.63-69
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• 2017

Golgi staining has been modified and developed since Camillo Golgi introduced the black reaction in 1873. This study focuses on the commonly used Golgi staining methods and presents comprehensive data regarding three Golgi staining methods along with their strong and weak points. The Golgi-Cox method uses mercuric chloride for brain tissue impregnation and is a reliable technique for analyzing the complete dendritic tree of cortical neurons. However, specimens tend to shrink during the staining steps. Recent combination of the Golgi-Cox method and immunofluorescence provides additional options for neuroscientists. Rapid Golgi staining requires osmium tetroxide for the post-fixation process. It homogenously stains whole structures of neurons and provides their detailed anatomical morphology. This staining is influenced by the age of the specimen, temperature of the laboratory, and duration of each procedure. The Golgi-Kopsch method uses formaldehyde and glutaraldehyde instead of osmium tetroxide and can be used regardless of the age of the specimen and the duration after fixation. This method is suitable for research using human brain fixed for a long time or for specimens obtained from old-aged animals. Selecting a Golgi staining protocol that is appropriate for the specimen type and research purpose is important to achieve best results.

• 대한한방내과학회지
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• 제24권1호
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• pp.113-122
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• 2003

Objective : The aim of this study was to evaluate the efficacy of Gagamchunggan-tang on anti-inflammation reaction with lipopolysaccharide (LPS)-induced HepG2 cell. Method : We examined the effects of the Gagamchunggan-tang, a traditional drug for liver inflammation, on the process of lipopolysaccharide(LPS)-induced nuclear factor-${\kappa}Bp65(NF-{\kappa}Bp65)$ activation in HepG2 cell. SDS-PAGE, Western blotting, Immunofluorescence staining were studied. Results : Immunoblot analysis showed that the level of nucleic $NF-{\kappa}Bp65$ was rapidly up-regulated and cytosolic inhibitory $I-{\kappa}B{\alpha}$ was down-regulated by LPS challenge. While Gagamchunggan-tang inhibited an increase of $NF-{\kappa}Bp65$ and degradation of $I-{\kappa}B{\alpha}$ in HepG2 cell. Besides LPS-induced expression of a group of genes, such as tumor necrosis factor-${\alpha}(TNF-{\alpha})$, inducible nitric oxide synthase(iNOS) and cyclooxygenase-2 (COX-2), are repressed by Gagamchunggan-tang. It may be concluded that Gagamchunggan-tang attenuates the progress of LPS-induced inflammation by reduction of $NF-{\kappa}Bp65$ activation. Conclusion : The Gagamchunggan-tang would be useful as a therapeutic agent for endotoxin-induced liver disease.

• 대한수의학회지
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• 제29권1호
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• pp.27-35
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• 1989

To develop more specific and sensitive diagnostic methods for infectious bovine rhinotracheitis, 7-C-2 monoclonal antibody specific to polypeptides of infectious bovine rhinotracheitis virus (IBRV) was applied in indirect immunofluorescence antibody assay (IFA), indirect immunoperoxidase assay(IPA) and radial immunodiffusion enzyme assay (RIDEA). It was found that IBRV infected in MDBK cells could be detected as early as 8 hours post infection by IFA, and that IFA was more rapid and specific to identify IBRV antigen than IPA. The diagnostic efficacy of RIDEA and SN test was studied with 88 bovine sera. It was evident that RIDEA could eliminate the false positive reaction encountered in serum neutralization(SN) test, being more rapid and sensitive than the latter. Highly significant correlation coefficiency (r=0.76, p<0.01) was evaluated between the titers of sera and the diameters of RIDEA. Tracheal membranes and sera collected from 96 slaughtered cattle with lesions in respiratory organs were examined to detect IBRV antigen and antibody by IFA, RIDEA and SN test. It was presented that positive rates were 32.3% in IFA, 20.8% in RIDEA and 21.9% in SN test, and that coincidence rate between RIDEA and SN test were 100% in positive sera and 98.7% in negative sera. In conclusion, it was assumed that application of monoclonal antibody could improve the diagnostic efficacy of IBR by enhancing sensitivity and specificity of IPA, IFA and RIDEA.

• Journal of Genetic Medicine
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• 제12권1호
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• pp.31-37
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• 2015

Purpose: We investigated the neurogenic potentials of amniotic fluid-derived stem cells (AFSCs) according to the expression levels of stem cell markers and ingredients in the neural induction media. Materials and Methods: Four samples of AFSCs with different levels of Oct-4 and c-kit expression were differentiated neurally, using three kinds of induction media containing retinoic acid (RA) and/or a mixture of 3-isobutyl-1-methylxanthine/indomethacin/insulin (neuromix), and examined by immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR) for their expression of neurospecific markers. Results: The cells in neuromix-containing media displayed small nuclei and long processes that were characteristic of neural cells. RT-PCR analysis revealed that the number of neural markers showing upregulation was greater in cells cultured in the neuromix-containing media than in those cultured in RA-only medium. Neurospecific gene expression was also higher in Oct-4 and c-kit double-positive cells than in c-kit-low or -negative cells. Conclusion: The stem cell marker c-kit (rather than Oct-4) and the ingredient neuromix (rather than RA) exert greater effects on neurogenesis of AFSCs.

• 한국동물위생학회지
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• 제31권1호
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• pp.101-111
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• 2008

The purpose of this study was to investigate the protective effects against porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) in suckling piglets by oral administration of IgY. Twenty piglets were divided into two groups with the same number: group I (treated with IgY) and group II (not treated). Group I was administerd orally with IgY for three days from one-day-old and experimentally challenged with PEDV and TGEV at four-day-old. The other was administered with saline solution and challenged with PEDV and TGEV at four-day-old. Serum antibody titers against PEDV and TGEV were examined by enzyme-linked immunosorbent assay (ELISA) and the detection of PEDV or TGEV antigen from feces and small intestines was performed by reverse transcription-polymerase chain reaction (RT-PCR) and indirect immunofluorescence (IFA). The antibody titers of the group I was higher than that of the other, and lasted at the end of experiment. In the detection tests of both virus from feces and small intestine, the rate of the group I was lower. Based on these results, oral administration of IgY may be effective to prevent the diarrhea caused by PEDV and TGEV.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권3호
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• pp.239-247
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• 2005

This study demonstrated that xenogenic human marrow mesenchymal stem cells (hMSCs) could elicit the regeneration of the sensory nerve after axotomy in the adult rats infraorbital nerves without immunosuppression. For this, we evaluated the behavioral testing for functional recovery of the nerve and histological findings at weeks 3 and 5 compared to controls. Xenogenic hMSCs did not evoke any significant inflammatory or immunologic reaction after systemic and local administrations. HMSCs-treated rats exhibited significant improvement on sensory recovery tested with von Frey monofilaments. At 5 postoperative weeks, in the hMSCs treated nerve, expression of myelin basic protein (MBP), neurofilament (NF) at the site of axotomy was higher than control. And mRNA expression of neurotropin receptor Trk precursor (TrkPre), nerve growth factor receptor (NGFR) and neuropeptide (NPY) in trigeminal ganglion were also higher. The number of myelinated nerve at distal stump and cells in trigeminal ganglion were higher in hMSC treated rats. So it was supposed that transplanted MSCs contributed to reducing post-traumatic degeneration and production of neurotrophic factors. Immunofluorescence labeling showed small portion of hMSCs (<10%) expressed a phenotypic marker of Schwann cell (S-100). Xenogenic or allogenic mesenchymal stem cells might have immune privileged characteristics and useful tool for cell based nerve repair.

• Clinical and Experimental Reproductive Medicine
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• 제47권2호
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• pp.108-113
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• 2020

Objective: Endometrial fibrosis, the primary pathological feature of intrauterine adhesion, may lead to disruption of endometrial tissue structure, menstrual abnormalities, infertility, and recurrent pregnancy loss. At present, no ideal therapeutic strategy exists for this fibrotic disease. Eupatilin, a major pharmacologically active flavone from Artemisia, has been previously reported to act as a potent inducer of dedifferentiation of fibrotic tissue in the liver and lung. However, the effects of eupatilin on endometrial fibrosis have not yet been investigated. In this study, we present the first report on the impact of eupatilin treatment on transforming growth factor beta (TGF-β)-induced endometrial fibrosis. Methods: The efficacy of eupatilin on TGF-β-induced endometrial fibrosis was assessed by examining changes in morphology and the expression levels of fibrosis markers using immunofluorescence staining and quantitative real-time reverse-transcription polymerase chain reaction. Results: Eupatilin treatment significantly reduced the fibrotic activity of TGF-β-induced endometrial fibrosis in Ishikawa cells, which displayed more circular shapes and formed more colonies. Additionally, the effects of eupatilin on fibrotic markers including alpha-smooth muscle actin, hypoxia-inducible factor 1 alpha, collagen type I alpha 1 chain, and matrix metalloproteinase-2, were evaluated in TGF-β-induced endometrial fibrosis. The expression of these markers was highly upregulated by TGF-β pretreatment and recovered to the levels of control cells in response to eupatilin treatment. Conclusion: Our findings suggest that suppression of TGF-β-induced signaling by eupatilin might be an effective therapeutic strategy for the treatment of endometrial fibrosis.