• The Journal of Pediatrics of Korean Medicine
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• v.28 no.2
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• pp.23-39
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• 2014

Objectives PRAL (Prunus armniaca Linne Var) is a herbal formula in Oriental Medicine, known for its anti-inflammatory and anti-allergenic properties. However, its mechanism of action and the cellular targets have not yet been found enough. The purpose of this study is to investigate the effects of PRAL on Th2 cytokines expression in MC/9 mast cells. Methods The effect of PRAL was analyzed by ELISA, Real-time PCR, Western blot in MC/9 mast cells. mRNA levels of GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ were analyzed with Real-time PCR. Levels of IL-13, MIP-$1{\alpha}$ were measured using enzyme-linked immunosorbent assays (ELISA). NFAT, AP-1 and NF-${\kappa}B$ p65 were examined by Western blot analysis. Results PRAL inhibited GM-CSF, IL-4, IL-5, IL-6, IL-13, TNF-${\alpha}$ mRNA expression in a dose dependent manner. GM-CSF, IL-4, IL-5 mRNA expression were inhibited significantly in comparison to DNP-IgE control group at concentration of 100 ${\mu}g/ml$ and IL-6, IL-13, TNF-${\alpha}$ mRNA expression were inhibited at concentration of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$. PRAL also inhibited the IL-13, MIP-$1{\alpha}$ production significantly in comparison to DNP-IgE control group in a dose dependent manner. IL-13 production was inhibited at a concentration of 200 ${\mu}g/ml$, 400 ${\mu}g/ml$ and MIP-$1{\alpha}$ was inhibited at a concentration of 100 ${\mu}g/ml$, 200 ${\mu}g/ml$, 400 ${\mu}g/ml$. Western blot analysis of transcription factors involving Th2 cytokines expression revealed prominent decrease of the mast cell specific transcription factors including NFAT-1, c-Jun as well as NF-${\kappa}B$ p65 but not NFAT-2 and c-Fos. Conclusion These results indicate that PRAL has the effect of suppressing Th2 cytokines production in the MC/9 mast cells. These data represent that PRAL potentiates therapeutic activities to the allergic disease by regulating Th2 cytokines in the MC/9 mast cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.12 no.1
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• pp.72-81
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• 1990

Because failure of microvascular flap grafting has sometimes been attributed to vascular obstruction in the anastomotic site, this study investigated the healing process after microvascular anastomoses. The healing process of anastomotic sites were observed by the use of the light and the scanning electron microscope after microvascular anastomoses of the right common carotid artery in rats. The experimental animals were sacrificed on the 4th day, 1st, 2nd, 4th and 6th week. Throughout the whole experimental period, arterial patency rate was 78% (11/14). At the early stage, it was possible to recognize histologically disappearance of endothelial cell and rupture of part of the media. Subintimal hyperplasia and the growth of media appeared around the suture line at the 2nd week. Endothelial cell regeneration occurred and the depth of vessel wall was normalized at the 4th week. By the scanning electron microscope, at the early stage, the anastomotic site was covered with many platelets, red blood cells, fibrins and macrophages. At the 4th week, the insertion site of the thread was completely covered with normal endothelial cells which were parallel to longitudinal axis of vessels and complete reendothelialization over the anastomotic site seemed to take about 6th week.

• Herbal Formula Science
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• v.15 no.2
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• pp.147-160
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• 2007

Objectives: This study is carried out to investigate the anti-stress effects of the sihoueokgantang in rats. Methods: we performed the passive avoidance test and the forced swimming test. The degrees of TH were also measured with immunohistochemical method in the regions of VTA and LC. Results: 1. As the results of the passive avoidance test. the sihoueokgantang showed tendency to improvement in compared with control group. 2. As the results of the forced swimming test, immobility of rats was significantly decreased in sihoueokgantang in compared with control group. 3. As the results of TH expression in the region of the LC, TH-immunoreactive cells were significantly decreased in the sihoueokgantang 100 mg/kg. As the results of TH expression in the region of the VTA. TH-immunoreactive cells were significantly decreased in the sihoueokgantang 400 mg/kg. Conclusion: As the results above, it can be considered that sihoueokgantang has the effects of preventing depression and memory decrease caused by stress.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.7
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• pp.1021-1028
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• 2012

Peripheral blood mononuclear cells (PBMCs) discriminate microbial pathogens and induce T-cell responses of appropriate effector phenotype accordingly. Toll-like receptors (TLRs), in part, mediate this microbial recognition and differentiation while the development of T-cell effector functions critically depends on the release of Th1- or Th2- type cytokines. In the present study, buffalo PBMCs were stimulated under in vitro culture conditions by Bacillus subtilis cell wall petidoglycan, a TLR2 ligand, in a dose- and time- dependent manner. The expression of TLR2 as well as the subsequent differential induction of the Th1 and Th2 type cytokines was measured. Stimulation was analyzed across five doses of peptidoglycan ($10{\mu}g/ml$, $20{\mu}g/ml$, $30{\mu}g/ml$, $40{\mu}g/ml$ and $50{\mu}g/ml$) for 3 h, 12 h, 24 h and 36 h incubation periods. We observed the induction of TLR2 expression in a dose- and time-dependent manner and the peptidoglycan induced tolerance beyond $30{\mu}g/ml$ dose at all incubation periods. The correlation between peptidoglycan stimulation and TLR2 induction was found positive at all doses and for all incubation periods. Increased production of all the cytokines was observed at low doses for 3 h incubation, but the expression of IL-4 was relatively higher than IL-12 at the higher antigen doses, indicating tailoring towards Th2 response. At 12 h incubation, there was a pronounced decrease in IL-4 and IL-10 expression relative to IL-12 in a dose- dependent manner, indicating skewing to Th1 polarization. The expression of IL-12 was highest for all doses across all the incubation intervals at 24 h incubation, indicating Th1 polarization. The relative expression of TNF-${\alpha}$ and IFN-${\gamma}$ was also higher while that of IL-4 and IL-10 showed a decrease. For 36 h incubation, at low doses, relative increase in the expression of IL-4 and IL-10 was observed which decreased at higher doses, as did the expression of all other cytokines. The exhaustion of cytokine production at 36 h indicated that PBMCs became refractory to further stimulation. It can be concluded from this study that the cytokine response to sPGN initially was of Th2 type which skews, more pronouncedly, to Th1 type with time till the cells become refractory to further stimulation.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.211-218
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• 2001

The $M_r$ 63,000 glycoprotein (GP63) and lipophosphoglycan (LPG) of Leishmania donovani were evaluated as vaccine candidates against visceral leishmaniasis. Mice were immunized with liposomeencapsulated GP63 and/or LPG that were purified from the soluble extract of L. donovani promastigotes, and were challenged with virulent amastigotes. Mice immunized with GP63/LPG in liposomes plus BCG resulted in a 27.4% reduction of amastigotes in the liver compared to the control group (liposomes plus BCG), and mice immunized with liposome-GP63 plus BCG failed to induce a protective immune response against the challenge infection. Immunization of mice with GP63 fused to the Schistosoma japonicum glutathione S-transferase (GP63-GST) plus BCG also failed to elicit protective immunity. To analyze the cause of failure to induce protection, cytokine release from the spleen cells of immunized mice and Leishmania-specific serum antibodies were analyzed. Spleen cells from mice immunized with GP63-GST plus BCG that were exposed to soluble extract of L. donovani in vitro produced 10-fold greater quantities of IFN-gamma and 3-fold greater quantities of IL-5 than cells from mice receiving BCG only or saline. Western blot analysis revealed that sera from these mice had Leishmania-specific antibodies recognizing 1 to 3 antigens of L. donovani with M. W. of 60-65 kDa. Although immunization of mice with GP63-GST induced a strong Th1 response, this study indicated that GP63-GST simultaneously elicited the Th2 response of the CD4+ T-cell, which was known to abrogate the protective immune response conferred by the Th1 effector function.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.1
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• pp.41-49
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• 2009

Soeumin-Googwihyangso-san(SGGHSS) has been used for the prevention or treatment of Soeumin-allergic rhinitis. This study was performed to demonstrate anti-allergic and anti-inflamatory effects of SGGHSS methanol extract in HMC cell lines and activated mouse B cells and CD4+ T cells. SGGHSS inhibited the production of TNF-$\alpha$ in PMA plus A23187 activated HMC-1 cells but not that of IL-6, as measured by ELISA. SGGHSS inhibited the expression of CD23 and surface IgE in B cells as determined by flowcytometry. It also inhibited secretion of IFN-$\gamma$ and IgG1, the Th1 related IgG type, but increased that of IL-4 in anti-CD40 and IL-4 treated B cells as measured by ELISA. As for Th cell differentiation, SGGHSS did not much affect IL-4 or IFN-$\gamma$. Taken together, our data showed that SGGHSS exerted an anti-inflammtory effect by inhibiting TNF-$\alpha$ in mast cells and has anti-allergic activity not via inhibition of CD4+ T cell, but via inhibition of B cells. These results suggest some evidence that SGGHSS can be applied to allergic disease.

• The Korea Journal of Herbology
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• v.29 no.6
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• pp.27-33
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• 2014

Objectives : The aims of this study were to exploring the therapeutic effect of Imperatae Rhizoma Extract(IRE) on Asthma. Methods : To investigate biological modulation activities of IRE, we conducted the cell-based assay whether IRE could regulate T helper 2 cells activity with EL4 T cells and mouse splenocytes, and followed animal study to conform the efficacy of their therapeutic potential on OVA-induced asthmatic mouse. Results : In cell study, IRE suppress the nuclear translocation of GATA binding protein-3 protein in phorbol 12-myristate 13-acetate/Ionomycin-stimulated EL4 T cells and Interleukine(IL)-4, IL-5 and IL-13 production in splenocytes at concentration dependent manner. In animal study, IRE-treated groups both 100mg/ml and 200mg/ml improve airway hypersensitibility reaction(AHR) response to methacholine about 30% and 40% with positive control group. Peritoneal blood analysis reveal that eosinophil number and ovalbumin-specific IgE is reduced by IRE treatment. Cell number of eosinophil is also reduced in bronchoalveolar lavage of IRE group like to peritoneal cell and real time-polymerase chain reaction data show that expression levels of IL-4, IL-5 and IL-13 were down regulated in lung tissue. Finally, histological analysis indicate that IRE protect the bronchial tissue damages through the accumulation of inflammatory cells and collagen, and these effect may be cause by interfering Th2 cells activity. Conclusions : Our data represent that IRE potentiates therapeutic activities to the allergic diseases such as asthma by regulating Th2 cells differentiation.

• Molecules and Cells
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• v.24 no.1
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• pp.132-138
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• 2007

4-1BB (CD137), a member of the tumor necrosis factor receptor superfamily, is expressed on activated T-cells, and 4-1BB signaling due to interaction with 4-1BB ligand or ligation with anti-4-1BB monoclonal antibody (mAb) costimulates T cells. It has been shown that administration of anti-4-1BB mAb induces anti-tumor immunity in mice, but the nature of the cellular subsets responsible for this immunity is uncertain. In this study we found that anti-4-1BB mAb administration to B16F10 melanoma-bearing mice induced marked expansion of $CD11c^+CD8^+$ T-cells in parallel with suppression of pulmonary tumors. The mAb-treated mice produced higher levels of $IFN-{\gamma}$ in their tumor tissues, spleen and lymph nodes than mice exposed to control antibody. When the $CD11c^+CD8^+$ T-cells were purified and re-stimulated in vitro, they produced high levels of the Th1 cytokines, $IFN-{\gamma}$ and IL-2, but low levels of the Th2 cytokines, IL-4 and IL-10. Furthermore, they expressed high levels of 4-1BB and CD107a, a marker of activated cytotoxic T-lymphocytes. Our results suggest that $CD11c^+CD8^+$ T-cells play a role in the anti-tumor immunity induced by anti-4-1BB mAb.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.232-238
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• 2020

This study used adult wistar-based rats to observe the sexual cycle as a morphological characteristic of vaginal epithelial cells by vaginal smearing, and investigated the fetal number through mating with male rats of the same strain. The target animal was a 12 to 13-week-old Wistar-based mature unlighted rat (weight 220 g to 240 g), room temperature 23 ± 2℃, 14 hours artificial lighting (05:00 to 19:00 hours), 10 hours Adapted individuals were used for rearing for at least 2 weeks under the conditions of the darkroom (19:00 to 05:00). The feed was managed for free feeding of pellet feed for animals and water. The vaginal smearing method was used for the experiments by observing the sexual cycle every morning and confirming that the normal sexual cycle of 4 or 5 days was repeated at least 2 cycles or more. As a result, the proestrus was found to have few red blood cells, the cells and nuclei were rather large and round, and many nucleated cells were identified. In the case of the estrus, the cells were large and the nuclei were not stained, and most of the keratinocytes were found. In addition, in the metestrus and diestrus, there were many white blood cells, and it was confirmed that nucleated epithelial cells and keratinocytes were significantly reduced. The pregnancy period was 21 ± 1.8 days, and the number of live births per delivery was 11.9 on average. The number of fetuses on the 8th and 10th days of pregnancy were 15.2 ± 0.4 and 15.4 ± 0.3, respectively. On the contrary, the number of fetuses on the 12th day of pregnancy was 12.9 ± 0.6, which was significantly (p < 0.05) decreased compared to the 10th day of pregnancy, and the number of fetuses was similar until delivery. As a result of investigating the change of body weight according to the birth weight and growth stage after delivery, the birth weight of female and male was 9.2 ± 2.0 g and 9.8 ± 2.5 g, respectively. After that, until the 16th day, the female and the male showed similarly moderate weight gain, and then showed a rapid weight gain until the 21st day of lactation. With reference to the results of this study, it is expected to be used as basic data for determining the mating time of rodents and controlling pregnancy and fetal number.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.13 no.3
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• pp.252-264
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• 1991

The purpose of this study was to compare the response of adjacent tissue and new bone formation after implantation by different methods of subperiosteal using using Proplast I and II in rabbit mandible. Microstructure of Proplast I and II was observed by scanning electron microscope. And the implantation procedure was carried out by dividing into tow groups, A and B. a group consisted of subperiosteal graft on the cortex, and the other B group was made up onlay graft following artificial decortication in the madibular body of rabbit. The experimental animals were sacrificed on the 1st, 2nd, 4th and 8th week after grafting for macroscopic and histopathologic examination. The samples extracted at the 6th postgrafting week were also used for biometric test. The result ere as follows : 1. By scanning electron microscopic observation, pore size was $50{\sim}180{\mu}m$ in the Proplast I and $100{\sim}220{\mu}m$ in Proplast II. 2. Macroscopically, infection of the graft site, deformation and displacement of the implanted materials were not observed in all experimental groups. 3. In the tissue response, infiltration of inflammatory cells and multinucleated giant cells were observed from the 2nd to the 8th week in Proplast I. Inflammatory cells decreased in number from the 2nd week in Proplast II suggesting that Proplast II is better than Proplast I. 4. Bone formation was not observed until the 8th week in the group A, but new bone formation from the surrounding graft bed and the periostium was appeared from the 4th week in the group B. 5. The maximum mean values of shear stress mere serially $65.5gf/mm^2$ in Proplast II of group B, $32.9gf/mm^2$ in Proplast I of group B, $17.0gf/mm^2$ in Proplast II of group A, and $15.7gf/mm^2$ in Proplast I. of group A.