Atopic dermatitis (AD) is an inflammatory skin disease commonly characterized by infiltration of inflammatory cells into skin lesions. Keratinocytes produce many chemokines that are involved in the pathogenesis of skin disorders. In particular, macrophage-derived chemokine (MDC/CCL22) and thymus and activationregulated chemokine (TARC/CCL17) are Th2-type cytokines. Serum MDC and TARC levels are increased in AD patients. In this study, we investigated the anti-inflammatory effect and mechanism of action of the active fraction from Prunus yedoensis bark. We evaluated their inhibitory effects on the AD-like inflammatory markers (MDC and TARC) and JAK-STAT pathway (STAT1) in HaCaT keratinocytes. The EtOAc fraction of the crude extract (80% EtOH) and the E5 sub-fraction potently inhibited the induction of MDC and TARC mRNA and protein at 50 ${\mu}g$/mL in HaCaT cells. In addition, the E5 sub-fraction inhibited the phosphorylation of STAT1 protein associated with IFN-$\gamma$ signaling transduction in a dose-dependent manner. Thus, P. yedoensis may have antiatopic activity by suppressing the inflammatory chemokines (MDC and TARC).
This study was performed to evaluate the effects of platelet-derived growth factor (PDGF) and epidermal growth factor (EGF) on the characteristics of beagle dog's periodontal ligament (BPD) cells and bone marrow (BBM) cells which have the important role on the early stage of periodontal tissue regeneration in vitro. In control group, the cells ($1.5{\times}10^5$cells/ml) were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu]g/ml$ ascorbic acid, and 10mM/ml ${\beta}-glycerophosphate$. In experimental groups, growth factors, PDGF or EGF(10ng/ml), were added into the above culture condition. And then each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity at 1, 5, 9, 13, 17th day after seeding of cells into the culture wells. The results were as follows: 1. Both BPD and BBM cells in PDGF-treated group proliferated more rapidly than non-treated cells. This finding also was observed in EGF-treated group but it was not as prominent as that of PDGF-treated group. The proliferation rates of both cells showed the time-dependent pattern during experimental periods in all three groups. 2. Amount of total protein synthesis was more increased in PDGF-treated group than in control group. But no significant difference between EGF-treated group and control group was observed throughout experimental periods even though the tendency of amount of protein synthesis was time-dependent pattern. 3. Alkaline phosphatase activity also more increased in PDGF-treated group than control group. But slight decrease tendency was seen in both cells of EGF-treated group. From the above results, PDGF appeared to enhance the proliferation and cellular activities including amount of total protein synthesis and alkaline phosphatase activity of BPD and BBM cell, but EGF did not show notable effects. The optimal application of these growth factors was thought to be useful as the adjunctive means in periodontal regeneration procedures.
An, Tai Joon;Rhee, Chin Kook;Kim, Ji Hye;Lee, Young Rong;Chon, Jin Young;Park, Chan Kwon;Yoon, Hyoung Kyu
Tuberculosis and Respiratory Diseases
/
v.81
no.1
/
pp.80-87
/
2018
Background: Asthma is a disease of chronic airway inflammation with heterogeneous features. Neutrophilic asthma is corticosteroid-insensitive asthma related to absence or suppression of $T_H2$ process and increased $T_H1$ and/or $T_H17$ process. Macrolides are immunomodulatory drug that reduce airway inflammation, but their role in asthma is not fully known. The purpose of this study was to evaluate the role of macrolides in neutrophilic asthma and compare their effects with those of corticosteroids. Methods: C57BL/6 female mice were sensitized with ovalbumin (OVA) and lipopolysaccharides (LPS). Clarithromycin (CAM) and/or dexamethasone (DXM) were administered at days 14, 15, 21, 22, and 23. At day 24, the mice were sacrificed. Results: Airway resistance in the OVA+LPS exposed mice was elevated but was more attenuated after treatment with CAM+DXM compared with the monotherapy group (p<0.05 and p<0.01). In bronchoalveolar lavage fluid study, total cells and neutrophil counts in OVA+LPS mice were elevated but decreased after CAM+DXM treatment. In hematoxylin and eosin stain, the CAM+DXM-treated group showed less inflammation additively than the monotherapy group. There was less total protein, interleukin 17 (IL-17), interferon ${\gamma}$, and tumor necrosis factor ${\alpha}$ in the CAM+DXM group than in the monotherapy group (p<0.001, p<0.05, and p<0.001). More histone deacetylase 2 (HDAC2) activity was recovered in the DXM and CAM+DXM challenged groups than in the control group (p<0.05). Conclusion: Decreased IL-17 and recovered relative HDAC2 activity correlated with airway resistance and inflammation in a neutrophilic asthma mouse model. This result suggests macrolides as a potential corticosteroid-sparing agent in neutrophilic asthma.
Purpose : We describe the changes of rat glomerular epithelial cells when exposed to high levels of glucose and advanced glycosylation endproducts(AGE) in the in vitro diabetic condition. We expect morphological alteration of glomerular epithelial cells and permeability changes experimentally and we may correlate the results with a mechanism of proteinuria in DM. Methods : We made 0.2 M glucose-6-phsphate solution mixed with PBS(pH 7.4) containing 50 mg/mL BSA and pretense inhibitor for preparation of AGE. As control, we used BSA. We manufactured and symbolized five culture dishes as follows; B5 - normal glucose(5 mM) + BSA, B30 - high glucose(30 mM) + BSA, A5 - normal glucose(5 mM) + AGE, A30 - high glucose(30 mM) + AGE, A/B 25 - normal glucose(5 mM) + 25 mM of mannitol(osmotic control). After the incubation period of both two days and seven days, we measured the amount of heparan sulfate proteoglycan(HSPG) in each dish by ELISA and compared them with the B5 dish at 2nd and 7th incubation days. We observed the morphological changes of epithelial cells in each culture dish using scanning electron microscopy(SEM). We tried the permeability assay of glomerular epithelial cells using cellulose semi-permeable membrane measuring the amount of filtered BSA through the apical chamber for 2 hours by sandwich ELISA. Results : On the 2nd incubation day, there was no significant difference in the amount of HSPG between the 5 culture dishes. But on the 7th incubation day, the amount of HSPG increased by 10% compared with the B5 dish on the 2nd day except the A30 dish(P<0.05). Compared with the B5 dish on the 7th day the amount of HSPG in A30 and B30 dish decreased to 77.8% and 95.3% of baseline, respectively(P>0.05). In the osmotic control group (A/B 25) no significant correlation was observed. On the SEM, we could see the separated intercellular junction and fused microvilli of glomerular epithelial cells in the culture dishes where AGE was added. The permeability of BSA increased by 19% only in the A30 dish on the 7th day compared with B5 dish on the 7th day in the permeability assay(P<0.05). Conclusion: We observed not only the role of a high level of glucose and AGE in decreasing the production of HSPG of glomerular epithelial cells in vitro, but also their additive effect. However, the role of AGE is greater than that of glucose. These results seems to correlate with the defects in charge selective barrier. Morphological changes of the disruption of intercellular junction and fused microvilli of glomerular epithelial cells seem to correlate with the defects in size-selective barrier. Therefore, we can explain the increased permeability of glomerular epithelial units in the in vitro diabetic condition.
We have reported that a 24 kDa protein (22U homologous; As22U) of Anisakis simplex larvae could elicit several Th2-related chemokine gene expressions in the intestinal epithelial cell line which means that As22U may play a role as an allergen. In order to determine the contribution of As22U to allergic reactions, we treated mice with 6 times intra-nasal application of recombinant As22U (rAs22U). In the group challenged with rAs22U and ovalbumin (OVA), the number of eosinophils in the bronchial alveolar lavage fluid (BALF) was significantly increased, as compared to the group receiving only OVA. In addition, mice treated with rAs22U and OVA showed significantly increased airway hyperresponsiveness. Thus, severe inflammation around the airway and immune cell recruitment was observed in mice treated with rAs22U plus OVA. The levels of IL-4, IL-5, and IL-13 cytokines in the BALF increased significantly after treatment with rAs22U and OVA. Similarly, the levels of anti-OVA specific lgE and lgG1 increased in mice treated with rAs22U and OVA, compared to those treated only with OVA. The Gro-${\alpha}$ (CXCL1) gene expression in mouse lung epithelial cells increased instantly after treatment with rAs22U, and allergy-specific chemokines eotaxin (CCL11) and thymus-and-activation-regulated-chemokine (CCL17) gene expressions significantly increased at 6 hr after treatment. In conclusion, rAs22U may induce airway allergic inflammation, as the result of enhanced Th2 and Th17 responses.
We describe here a case of malignant mixed osteogenic tumor of the mammary gland with alveolar carcinomatous appreance. A firm, 2 to 2.5cm (in diameter) mass under the 5th nipple, showing the structure of extraosseous osteogenic sarcoma, was removed from the left 5th mammary gland of 12-year-old female dog. When investigated under the microscope, the osteoid material undergoing mineralization was surrounded by numerous scattered osteoblasts and a few osteoclastic cells throughout the osteoid tumorous stroma. The osteoid lesions were continuous with hypercellular myoepithelial cells of a very immature character with several mitotic figures. In addition, there were also carcinomatous tubules and alveoli, with invading cells into peripheral stroma, surrounded by myoepithelial cells in the mammary gland. In these lesions, emanating cords of tumor cells appear to be continuous with the myoepithelial cell layer of a duct. The presence of all these cell types suggests the existence of a common malignant origin, the stem cell being differentiated into epithelial carcinomatous and mesenchymal sarcomatous chondral and osteogenic tissues.
Seo, Kil Woong;Kim, Chong Sup;Park, Chang Sik;Lee, Kyu Seung
Korean Journal of Agricultural Science
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v.17
no.1
/
pp.34-44
/
1990
The study was carried out to elucidate the effects of ovarian function on the thyroid gland, adrenal gland and uterus in female rats. One hundred and forty-four mature female rats were allotted into the three groups ; ovariectomized group, estradiol treated group and intact control group. The ovaries of 48 heads of rats were completely removed. Forty eight heads of rats were administered with $200{\mu}g$ of estradiol benzoate every 48 hours. Serum estradiol-$17{\beta}$ and progesterone levels were determined with radioimmunoassay method at 3, 6, 12, 24 hours and 5, 10, 15 days after treatment. The rats were necropsied to measure weights of thyroid gland, adrenal gland and uterus and to examine the histological changes in the organs. The results obtained were as follows ; 1. Serum estradiol-$17{\beta}$ levels were rapidly decreased below 27.20pg/ml 18 hours after ovariectomy. In estradiol treated rats the levels were rapidly increased 18 hours after treatment, but thereafter slowly decreased. The significant differences in the estradiol level were found between the group at every observation time. 2. Serum progesterone levels were significantly decreased after ovariectomy and estradiol injection. The lowest level was found in the group of ovariectomized rats. 3. The weights of thyroid glands decreased in ovariectomized rats rather than in intact rats 5 days after treatment. The weights tended to increase after estradiol injection but significant differences between the groups were seen on 10th and 15th days. 4. In the histological findings of thyroid glands, follicular epithelial cells were changed to be squamous 5 days after ovariectomy and accompanied pyknosis 10 days and karyorrhexis 15 days after ovariectomy. On the contrary follicular epithelial cells were changed to be columnar with hypertrophy 10 days after estradiol injection. 5. The significant differences in adrenal gland weights were recognized between all the groups 5 and 15 days after treatment in ovariectomized rats were lighter than intact rats and the adrenal gland weights were rather heavier in estradiol treated rats. 6. The days after ovariectomy the adrenal glands were atrophied accompanying with pyknosis in the cortical cells of zona fasciculata. The cells in zona fasciculata and zona reticularis started to hypertrophy 5 days after estradiol injection, but no changes were found in the zona glomerulosa of adrenal cortex and in the adrenal medulla. 7. The significant differences in uterus weights were recognized between the groups at each observation time. After ovariectomy the uterus weights decreased rapidly but after estradiol injection they increased rapidly. 8. Through histological examination of uterus, the atrophy and degeneration started to occur in endometrium and lamina propria 12 hours after ovariectomy, and in myometrium one day after ovariectomy, and the changes progressed rapidly after that. On the contrary, the myometrium was proliferated and hypertrophied from 12 hours after estradiol-$17{\beta}$ injection.
In order to investigate the antitumor effect by Chungsangbohahwan after B-16 cells were transplanted in C57BL/6 mice, and the immune responses in mice induced by methotrexate, the extract of Chungsangbohahwan was orally administered to the mice for 21 days. Experimental studies were performed for measurance of metastasis, cell cytotoxicity in vitro, natural killer cell activity, productivity of interleukin-2. The results were summarized as follows: 1. Inhibition of metastasis in Chungsangbohahwan-treated group was higher than control group with significance on 7th day and 14th day. 2. On the MTT assay, cell viability was significantly inhibited by $5{\mu}g/well$, $2.5{\mu}g/well$, $1.25{\mu}g/well$, and $0.625{\mu}g/well$ of Chungsangbohahwan concentration inhibited cell viability significantly(P<0.05). $IC_{50}$ for cell viability was $2.17{\mu}g/well$. 3. Natural killer cell activity in Chungsangbohahwan-treated group was significantly increased on 100:1, 50:1 E/T(effect cell/target cell) ratio(P<0.05). 4. Production of interleukin-2 in Chungsangbohahwan-treated group was significantly increased(P<0.05).
Unlike most bacteria, Treponema pallidum subspecies cannot be readily isolated or sustained in cell culture for numerous generations. In korea, two non treponemal tests are currently considered as standard; the VDRL slide test and RPR card test. These tests are based on an antigen composed of an alcoholic solution containing measured amount of cardiolipin, cholesterol, and sufficient purified lecithin to produce reactivity. The nontreponemal reagin tests measure immunoglobulin M (IgM) and IgG to lipoidal material released from damaged host cells as well as to lipoprotein-like material and possibly by cardiolipin released from the treponemes. The object of the evaluation was to evaluate the performance of the Mediace RPR kit on the automated biochemistry analyzer system as a method for screen method of syphilis as well as to identify BFP possibility. For evaluation of routine screening test, a total 2,380 specimens tested by Mediace RPR from 28th Oct, 2007 to 22th Feb, 2008. For evaluation of BFP possiblility, we measured samples which have potential BFP reaction in Syphilis test such as ANA (anti-nuclear antibody) positive (135 samples), CRP (C-reactive protein) positive (100 samples), RF (Rheumatoid factor) positive (26 samples), and other potential BFP cases (17 samples) including total 278 samples. These samples were tested quantitative test Mediace RPR with Hitachi 7600 P module. For comparison with current manual test, VDRL slide test were performed. Of these 2380 specimens, 2350 were negative, 30 were positive, and one were positive with TPHA. Both methods agreed for 2356 (98.9%) samples. Of the 30 samples showed positive results over 1.0 R.U, 6 samples showed positive results with VDRL test. Of these 6 samples, 1 samples showed positive with TPHA test. The combination of the Automated Biochemistry analyzer and VDRL test for retest can be increase efficiency of syphilis screening test.
We investigated the hypoglycemic effect of formula containing Euonymus alatus (EA) and Mori Folium (MF) in multiple low dose (MLD) streptozotocin (STZ)-induced diabetic rats. In order to iduce hyperglycemic state 25 mg/kg of STZ was injected intraperitoneally for 5 consecutive days. SD rats were randomly divided into diabetic control and treatment groups. Treatment groups were administered with either 250 mg/kg of EA and 250 mg/kg of MF (E1Ml), or 500 mg/kg of EA mixed with same dose of MF (E2M2) for 3 weeks. Blood glucose levels and body weights were measured every 5th or 6th day. E1Ml and E2M2 both significantly reduced food intake, water intake, and fasting blood and urine glucose levels as compared to those in diabetic control group in a dose dependent manner. Body weight in diabetic control group was increased slightly after 3 weeks. Treatment group, however, showed gradual increase in body weights during 3 week-period. While plasma insulin levels of the diabetic control group were decreased to the level of 387$\pm$14 pg/ml from 534$\pm$36 pg/ml, those levels in E1Ml and E2M2-treated groups were both markedly increased by 13% and 26%, respectively. Urine glucose levels in E1Ml and E2M2-treated groups were also remarkably reduced by 17 and 26% compared to the levels of diabetic control group. While expression of membrane-bound glucose transporter-4 (GLUT-4) protein in skeletal muscle was reduced by 45% in diabetic control compared to the normal control, GLUT-4 protein expressions in E1Ml and E2M2-treated groups were augmented by 2 and 3.5 times compared to the diabetic control, respectively. Pancreatic HE staining experiments showed that E2M2-treated group revealed much less infiltrated mononuclear cells, indicating that E2M2 efficiently blocked insulitis induced by multiple low dose streptozotocin. Taken together, we conclude that formula containing EA and MF may prevent or delay the development of hyperglycemia through overexpression of GLUT-4 protein in skeletal muscle and prevention of insulitis.
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