Ferulic Acid (FA) is a metabolite of phenylalanine and tyrosine, a phenolic compound commonly found in fruits and vegetables. Several studies have shown that FA has various functions such as antioxidant effect, prevention of cell damage from irradiation, protection from cell damage caused by oxygen deficiency, anti-inflammatory action, anti-aging action, liver protective effect and anti-cancer action. In this study, we investigated the maturation rate, intracellular glutathione (GSH) and reactive oxygen species (ROS) of porcine oocytes by adding FA to the in vitro maturation (IVM) medium and examined subsequent embryonic developmental competence at 5% oxygen through parthenogenesis. There is no significant difference between the control group ($0{\mu}M$) and treatment groups ($5{\mu}M$, $10{\mu}M$, $20{\mu}M$) on maturation rates. Intracellular GSH levels in oocyte treated with $5{\mu}M$ of FA significantly increased (P < 0.05), and $20{\mu}M$ of FA revealed significant decrease (P < 0.05) in intracellular ROS levels compared with the control group. Oocytes treated with FA exhibited significantly higher cleavage rates (79.01% vs 89.19%, 92.20%, 90.89%, respectively) than the control group. Oocytes treated with $10{\mu}M$ showed significantly higher blastocyst formation rates (28.3% vs 40.3%, respectively) after PA than the control group. Total cell numbers in blastocyst of $10{\mu}M$ FA displayed significantly higher (39.4 vs 51.9, respectively) than the control group. In conclusion, these results suggested that treatment with FA during IVM improved the developmental potential of porcine embryos by increasing intracellular GSH synthesis and reducing ROS levels. Also, there was an improvement of cleavage rate, blastocyst formation and total cell numbers in blastocysts. It might be associated with Keap1-Nrf2 pathway as an antioxidant regulate pathway that plays a crucial role in determining the sensitivity of cells to oxidative damages by regulating the basal and inducible expression of enzymes which is related to detoxification and anti-oxidative effects, stress response enzymes and/or proteins and ABC transporters.
Atopic dermatitis is a chronic inflammatory skin disease caused by a variety of genetic and environmental factors, dysregulation of immunological response, as well as dysfunction of the skin barrier proteins. The purpose of this study is to develop an ELISA kit suitable for evaluating the expression of skin barrier proteins. Proteins were obtained from the skin via AriNo and D-Squame patches. The efficiency of protein collection from the skin, using the Arino patch, was shown to be more effective than using D-Squame; while the efficiency of lysis using 0.1% Triton-X100 was higher than that of other lysis solutions, including 0.1 M Tris-HCL, 0.1% Tween-20, and 5 mM KOH. Recombinant skin barrier proteins, such as filaggrin and involucrin, were produced by molecular biological methods. Monoclonal antibodies against filaggrin and involucrin were produced by immunization of mice, fusion of spleen cells and myeloma cells, as well as a selection of antibody-producing hybridoma cells. The filaggrin expression in the skin of subjects suffering from atopic dermatitis was lower than that in normal mice. Involucrin expression was not altered between normal individuals and subjects with atopic dermatitis. These findings contribute to an elucidation of the importance of the skin barrier protein expression in atopic dermatitis and the development of a diagnostic kit for atopic dermatitis.
Purpose: The most effective methods of harvesting, preparing, and injecting autologous fat grafts have been inconsistent and conflicting. With its limitation as resorption in fat grafting, handling various techniques affect adipocyte survival, and is crucial to optimizing its long-term survival. To improve graft survival, re-implantation of cryopreserved adipocytes was developed. In addition, adipocytes do not induce immune rejection in response to non-self lymphocytes in a mixed lymphocyte reaction. The purpose of this study is to analyze the changes in cryopreserved adipocytes so as to determine the most efficient long-term storage period, and to analyze the changes in cryopreserved allografted adipocytes so as to determine the efficacy of cryopreserved adipocytes allografting. Methods: Fat tissues were harvested from the inguinal and retroperitoneal fat pad of mice. After the centrifugation of the harvested fat tissues, they were disintegrated with collagenase. The adipocytes were obtained by centrifugation of the disintegrated fat tissues. The adipocytes were treated as follows: (1) They were examined for weight and then frozen at $-20^{\circ}C$(n=25). For four months, each five frozen samples were taken and examined for weight and histologic changes in the 1st week, the 1st month, the 2nd month, the 3rd month, and the 4th month, respectively. (2) The adipocytes were immediately frozen at $-20^{\circ}C$(n=125). For four months, five frozen samples were taken, and allografted in the same time period as above. Finally, for four months, five cryopreserved allografted adipocytes were taken and examined for histologic changes in the same time period as above. Results: (1) Significant weight changes and histologic findings with inflammatory and destructive changes were observed in the cryopreserved adipocytes in three months. (2) Significant fat necrotic changes in the histologic changes with Hematoxylin and eosin stain were observed in the cryopreserved allografted adipocytes since the first week, independent of the freezing period. Conclusion: The study results show that the adipocytes that were cryopreserved for more than three months underwent obvious weight reductions and necrotic changes, and the adipocytes that were allografted without freezing were viable for four months, but the cryopreserved allografted adipocytes had obvious necrotic changes since the first week regardless of the freezing period.
Journal of Physiology & Pathology in Korean Medicine
/
v.24
no.2
/
pp.290-295
/
2010
Gamigunggui-tang (GMGGT) is one of the important prescriptions that has been used in oriental medicine. We investigated the inhibitory effect of an oral administration of hot water extract of GMGGT on the development of atopic dermatitis (AD) by using Balb/c mice. The induction of atopic dermatitis-like lesion was conducted by the removal of the back hairs and topical application of 2,4-Dinitrochlorobenzene (DNCB) on to the back skin repeatedly. GMGGT was orally administered at a different dose (10.0 mg/kg, 50.0 mg/kg). Skin reactions, consisting of increased ear thickness and the presence of ear inflammation, were observed in mice over three weeks. Oral administration of GMGGT significantly suppressed the skin lesions, ear swelling, spleen weight, total serum IgE in a concentration dependent manner, and also inhibited the infiltration of mast cells in the dorsal skin. In the present study, these results suggested that GMGGT extract inhibits inflammatory response atopic dermatitis. Therefore, GMGGT may be effective substances for the management of AD in human.
Oroxylin A is a flavone isolated from a medicinal herb reported to be effective in reducing the inflammatory and oxidative stresses. It also modulates the production of brain derived neurotrophic factor (BDNF) in cortical neurons by the transactivation of cAMP response element-binding protein (CREB). As a neurotrophin, BDNF plays roles in neuronal development, differentiation, synaptogenesis, and neural protection from the harmful stimuli. Adenosine $A2_A$ receptor colocalized with BDNF in brain and the functional interaction between $A2_A$ receptor stimulation and BDNF action has been suggested. In this study, we investigated the possibility that oroxylin A modulates BDNF production in cortical neuron through the regulation of $A2_A$ receptor system. As expected, CGS21680 ($A2_A$ receptor agonist) induced BDNF expression and release, however, an antagonist, ZM241385, prevented oroxylin A-induced increase in BDNF production. Oroxylin A activated the PI3K-Akt-GSK-$3{\beta}$ signaling pathway, which is inhibited by ZM241385 and the blockade of the signaling pathway abolished the increase in BDNF production. The physiological roles of oroxylin A-induced BDNF production were demonstrated by the increased neurite extension as well as synapse formation from neurons. Overall, oroxylin A might regulate BDNF production in cortical neuron through $A2_A$ receptor stimulation, which promotes cellular survival, synapse formation and neurite extension.
This experiment was conducted to investigate effects of dietary protease on immune responses of weaned pigs. Weaned pigs (n = 75; 7.06 ± 0.18 kg BW; 28 d old) were randomly assigned to 3 treatments (5 pigs/pen; 5 pens/treatment). Dietary treatments were positive control, a diet with required protein level (PC), negative control, a diet with lower protein level than PC (NC), and NC + 0.02% dietary protease (PRO). The dietary protease used in this experiment was a commercial product containing 75,000 protease units/g derived from Nocardiopsis prasina produced in Bacillus licheniformis. The dietary treatments did not contain any ingredients or additives that may provide antibacterial or physiological effects. Pigs were fed respective dietary treatments for 6 weeks. Blood was collected from randomly selected 2 pigs in each pen on d 1, 3, 7, and 14 after weaning. Measurements were number of white blood cells (WBC), tumor necrosis factor-α (TNF-α), transforming growth factor-β1 (TGF-β1), and C-reactive protein (CRP). Pigs fed PRO had lower WBC on d 7 (14.84 vs 20.42 × 103/μL; p < 0.05) and TNF-α on d 7 (618 vs 889 pg/mL; p = 0.085) and 14 (437 vs 576 pg/mL; p = 0.069) than those fed NC, but there were no differences on WBC and TNF-α between PC and PRO. Pigs fed PRO had lower TGF-β1 on d 3 (630 vs. 1,588 and 1,396 pg/mL; p < 0.05) than those fed PC and NC. However, no differences were found on CRP among dietary treatments. In conclusion, addition of dietary protease reduced inflammatory immune responses of weaned pigs.
The in vivo and in vitro effects of oriental herb extracts of Cassia obtusifolia, Taraxacum platycarpum and Ulmusmacrocarpa on anti-atopic allergic reaction were evaluated in this study. A mixture of these extracts exhibited more potent anti-allergic activities in human mast cells than those from individual extracts. The herbal mixture significantly inhibited the release of compound 48/80-induced $\beta$-hexosaminidase release in the human mast cell line, HMC-1. The mixture also suppressed the production of PMA and A23187-induced inflammatory cytokines in HMC-1 cells. To further investigate the in vivo effects of the herbal mixture, a Dermatophagoides farinae (DF)-induced atopic dermatitis mouse model was utilized. Oral administration of the herbal mixture significantly decreased the ear thickness and swelling in DF treated NC/Nga mice in a dose dependent manner. Furthermore, serum levels of IgE and interleukin-4 (IL-4) were significantly decreased, whereas interferon-gamma (IFN-$\gamma$) levels were increased in the mixture administrated groups when compared to the control. Taken together, our data indicate the possibility of using a mixture of the oriental herb extract to relieve symptoms of atopic dermatitis.
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.33
no.2
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pp.114-120
/
2007
The augmentation of soft tissue defects is one of the critical problems in the oral and maxillofacial surgery. Various types of graft materials, both autologous and non-autologous, have been used for the augmentation of soft tissue in the facial region. However, it is not easy to choose an ideal material for soft tissue augmentation because each has its advantages and disadvantages. An ideal graft material should meet the following criteria : it should not leave a scar at the area from which it was taken; should have less likelihood of causing infection; should feel natural after implanted; and should be not absorbed. Among the materials meeting these criteria, human dermis and artificial dermis are commonly used for clinical purposes. The present study was aimed to investigate and compare the resorption rate and the histological change following the use of the autologous dermis, the human homogenous dermis $Alloderm^{(R)}$, and the artificial dermis $Terudermis^{(R)}$ to reconstruct the soft tissue defect. Twenty mature rabbits of either sex, weighing about 2 ㎏, were used. Each rabbit was transplanted with the autologous dermis, $Alloderm^{(R)}$, and $Terudermis^{(R)}$ size $1{\times}1-cm$ at the space between the external abdominal oblique muscle and the external abdominal oblique fascia. They were then divided into 4 groups (n=5 each) according to the time elapsed after the surgery: 1, 2, 4, and 8 weeks. The resorption rate was calculated by measuring the volume change before and after the transplantation, and H-E stain was preformed to observe the histological changes. The resorption rate after 8 weeks was 21.5% for the autologous dermis, 16.0% $Alloderm^{(R)}$, and 36.4% $Terudermis^{(R)}$, suggesting that $Alloderm^{(R)}$ is the most stable while $Terudermis^{(R)}$ is the most unstable. In microscopic examinations, the autologous dermis graft was surrounded by inflammatory cells and showed foreign body reactions. The epidermal inclusion cyst was observed in the autologous dermis graft. $Terudermis^{(R)}$ and $Alloderm^{(R)}$ demonstrated neovascularization and the progressive growth of new fibroblast. The results suggest that $Terudermis^{(R)}$ and $Alloderm^{(R)}$ can be availably for substituting the autologous dermis.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.16
no.3
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pp.116-128
/
2003
Objectives : The incidence of allergic rhinitis has increased but treatment in most cases has only dealt with the symptoms. Medicine has been developed that shows fewer side effects. However, some side effects and the psychological stress over taking medicine has remained. There have been no studies so far performed on the effect of this Jeulminmilmae-tang's use, only. The purpose of this study was find out the therapeutic effects of its exclusive use on an Animal Model with Allergic Rhinitis. Methods : Thirty Sprague-Dawley rats were divided into three group : normal group, control group and sample group. To induce the allergic rhinitis in control group and sample group, rats were sensitized intraperitoneally with 0.1$\%$ ovalumin solution 3 times at intervals of 1 week. Then intranasal sensitization was performed by diffusing 0.1$\%$ ovalumin solution 3 times at intervals of 2 days. After that time, rats in the sample group were oral administration treated by Jeulminmilmae-tang for 28 days. Compared with the sample group, rats in the control group were oral administration treated by normal saline for 28 days. We observed changes in nasal mucosa and submucosa; also changes in the segment of neutrophil, eosinophil, Iympocyte and monocyte in blood. We used the statistical methods of student t-test(p 〈0.05). And we observed the changes of AST, ALT of three groups and used anova test statistically. Results : The segment of eosinophil was significantly decreased in treated group when compared with the control group(p 〈0.05). The segment of neutrophil. in blood were decreased in the treated group when compared with the control group but. that was not significant statistically(p 〈0.05). There were some regrowth of the cilium in the treated group. Histologic changes showed edema congestion and expantion of grandular cells in nasal submucosa and hypertrophy of epithelium ill nasal mucosa were decreased in treated group when compared with control group. Effects of Jeulminmilmae-tang on the liver function were also studies in rats. Treatment of Jeulminmilmae-tang did not affected on AST and ALT. Conclusions : The results may suggest that oral administration treatment using Jeulminmilmae-tang decreases the inflammatory response on an Animal Model with Allergic Rhinitis.
Park, Jin-Hoon;Choi, Chung-Gon;Jeon, Sang-Ryong;Rhim, Seung-Chul;Kim, Chang-Jin;Roh, Sung-Woo
Journal of Korean Neurosurgical Society
/
v.49
no.5
/
pp.267-272
/
2011
Objective : Although iliac crest autograft is the gold standard for lumbar fusion, the morbidity of donor site leads us to find an alternatives to replace autologous bone graft. Ceramic-based synthetic bone grafts such as hydroxyapatite (HA) and b-tricalcium phosphate (b-TCP) provide scaffolds similar to those of autologous bone, are plentiful and inexpensive, and are not associated with donor morbidity. The present report describes the use of Polybone$^{(R)}$ (Kyungwon Medical, Korea), a beta-tricalcium phosphate, for lumbar posterolateral fusion and assesses clinical and radiological efficacy as a graft material. Methods : This study retrospectively analyzed data from 32 patients (11 men, 21 women) who underwent posterolateral fusion (PLF) using PolyBone$^{(R)}$ from January to August, 2008. Back and leg pain were assessed using a Numeric Rating Scale (NRS), and clinical outcome was assessed using the Oswestry Disability Index (ODI). Serial radiological X-ray follow up were done at 1, 3, 6 12 month. A computed tomography (CT) scan was done in 12 month. Radiological fusion was assessed using simple anterior-posterior (AP) X-rays and computed tomography (CT). The changes of radiodensity of fusion mass showed on the X-ray image were analyzed into 4 stages to assess PLF status. Results : The mean NRS scores for leg pain and back pain decreased over 12 months postoperatively, from 8.0 to 1.0 and from 6.7 to 1.7, respectively. The mean ODI score also decreased from 60.5 to 17.7. X-rays and CT showed that 25 cases had stage IV fusion bridges at 12 months postoperatively (83.3% success). The radiodensity of fusion mass on X-ray AP image significantly changed at 1 and 6 months. Conclusion: The present results indicate that the use of a mixture of local autologous bone and PolyBone$^{(R)}$ results in fusion rates comparable to those using autologous bone and has the advantage of reduced morbidity. In addition, the graft radiodensity ratio significantly changed at postoperative 1 and 6 months, possibly reflecting the inflammatory response and stabilization.
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