• Journal of Korean Medicine Rehabilitation
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• v.24 no.3
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• pp.51-69
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• 2014

Objectives This study was aimed to investigate the effects of Sibjeondaebotanggamibang (SJT) on the wound-induced rats. Methods It was observed the effects of anti-oxidation and anti-inflammation by using of lipopolysaccharide (LPS)-treated RAW 264.7 cells. For the observing on SJT anti-oxidation, it needed to mesure the total amount of polyphenol, DPPH scavenging ability, ABTS scavenging ability and the value of ROS production. In order to observe on the anti-inflammation of SJT, it was mesured the value of No and Cytokine (TNF-${\alpha}$, IL-$1{\beta}$, IL-6). It needed to make a scar (around $2{\times}2cm^2$) on the top of the fascia in the back of the rats and then the rats were divided into 4 groups (n=6). Control group was not treated at all, whereas SJ group was orally medicated SJT, Terra group was per-cutaneously applied Terramycin, and SJ+Terra group was both orally medicated SJT and percutaneously applied Terramycin per day for three weeks. The size of wound was measured with Digimatic Caliper and the blood samples (WBC, neutrophil, monocyte, lymphocyte) were analyzed using Minos-ST, which were collected by cardiac puncture. The effect on inflammatory cytokine (TNF-${\alpha}$, IL-$1{\beta}$, IL-6), immunological cells in synovial fluid was measured. To measure the wound factor expressed by wounded skin sample, we extracted RNA and to investigate MMP-1,2,9 we used RT-PCR. For performing histopathological examinations, we paralyzed the rats by ether, and extracted wounded skin tissues, which were measured by H & E, and monitored on the optical microscope. Results 1. DPPH and ABTS scavenging activity of SJT was increased concentration-dependantly, and ROS scavenging activity was significantly increased (10, $100{\mu}g/ml$). 2. NO production was significantly reduced in SJT treated cells ($100{\mu}g/ml$), both TNF-${\alpha}$ and IL-6 in SJT treated cells (1, 10, $100{\mu}g/ml$), and IL-$1{\beta}$ in SJT treated cells (1, $100{\mu}g/ml$). 1. The size of wound was significantly decreasing in SJ group, Terra group, SJ+Terra group. 2. WBC was significantly reduced in SJ and SJ+Terra group, monocyte in SJ+Terra group. Neutrophil was also reduced in SJ, SJ+Terra group but meaningless. 3. TNF-${\alpha}$ and IL-6 were significantly reduced in SJ group, Terra group, SJ+Terra group, and IL-$1{\beta}$ in SJ+Terra group. 4. mRNA expression in MMP-1 was significantly reduced in SJ group. 5. Collegan production and chronic inflammation were significantly decreased in SJ group, Terra group, SJ+Terra groups. Re-epithelization on the skin in Terra group, SJ+Terra groups was decreased. Conclusions According to this in vitro experiment, Sibjeondaebotanggamibang (SJT) has the effects of anti-oxidative and anti-inflammatory. By in vivo experiment, SJT has the effects of anti-inflammatory. Moreover, the progress of recovery was found visually, heamatologically, genetically and histopathologically. In conclusion, it could be thought that SJT has effect on the treating of wound.

• Tuberculosis and Respiratory Diseases
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• v.53 no.1
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• pp.5-16
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• 2002

Background : Though mononuclear phagocytes serve as the final effectors in killing intracellular Mycobacterium tuberculosis, the bacilli readily survive in the intracellular environment of resting cells. The mechanisms through which cellular activation results in the intracellular killing is unclear. In this study, we sought to explore an in vitro model of a low-level infection of human mononuclear phagocytes with MAC and $H_{37}Ra$ and determine the extent of the lymphocyte dependent cytotoxicity of human monocytes and alveolar macrophages. Materials and Methods : The peripheral monocytes were prepared using the Ficoll gradient method from PPD positive healthy people and tuberculosis patients. The alveolar macrophages were prepared from PPD positive healthy people via a bronchoalveolar lavage. The human mononuclear phagocytes were infected at a low infection rate (bacilli:phagocyte 1:10) with MAC(Mycobacterium avium) and Mycobacterium tuberculosis $H_{37}Ra$. Non-adherent cells(lymphocyte) were added at a 10:1 ratio. After 1,4, and 7 days culture in $37^{\circ}C$, 5% CO2 incubator, the cells were harvested and inoculated in a 7H10/OADC agar plate for the CFU assay. The bacilli were calculated with the CFU/$1{\times}10^6$ of the cells and the cytotoxicity was expressed as the log killing ratio. Results : The intracellular killing of MAC and $H_{37}Ra$ within the monocyte was greater in patients with tuberculosis compared to the PPD positive controls (p<0.05). Intracellular killing of MAC and $H_{37}Ra$ within the alveolar macrophage appeared to be greater than that within the monocytes of the PPD positive controls. There was significant lymphocyte dependent inhibition of intracellular growth of the mycobacteria within the monocytes in both the controls and tuberculosis patients and within the macrophages in the controls(p<0.05). There was no specific difference in the virulence between the MAC and the $H_{37}Ra$. Conclusion : This study is an in vitro model of a low-level infection with MAC and $H_{37}Ra$ of human mononuclear phagocytes. The intracellular cytotoxicity of the mycobacteria within the phagocytic cells was significantly lymphocyte dependent. During the 7 days culture after the intracellular phagocytosis, the actual confinement of the mycobacteria was observed within the monocytes of tuberculosis patients and the alveolar macrophages of the controls as in the case of adding lymphocytes.

• Tuberculosis and Respiratory Diseases
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• v.47 no.1
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• pp.13-25
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• 1999

Background: Ineffective cell-mediated immune response in human tuberculosis is associated with a depressed Thl cytokine response and reduced production of IFN-$\gamma$. Most persons infected with Mycobacterium tuberculosis are healthy tuberculin reactors with protective immunity, but a minority with ineffective immunity develop extensive pulmonary tuberculosis. The cell-mediated immune response is an important aspect of host resistance to mycobacterial infection and is believed to be tightly regulated by a balance between Th1 cytokines including IFN-$\gamma$, IL-12, IL-18, regulated on activation, normal T cell expressed and secreted (RANTES) and Th2 counterparts such as IL-4, monocyte chemoattractant protein-l (MCP-l). Methods: Proliferation and mRNA expression of IFN-$\gamma$, RANTES and MCP-l by RT-PCR in peripheral blood mononuclear cells (PBMCs) in response to in vitro stimulation with mycobacterial antigens were compared in pulmonary tuberculosis patients with cured and treatment failure and in tuberculin-positive and tuberculin-negative healthy subjects. Results: Defective proliferative responsiveness to aqueous TSP antigen was involved with treatment failure tuberculosis patients. Aqueous TSP antigen-induced IFN-$\gamma$ and RANTES mRNA expression was decreased in treatment failure tuberculosis patients compared with healthy tuberculin reactors and cured tuberculosis patients (23.1 % versus 90.0% for IFN-$\gamma$ and 46.2% versus 70.0% versus 46.2% for RANTES). The frequency of MCP-l mRNA expression to aqueous TSP antigen in treatment failure tuberculosis patients was greater than in healthy tuberculin reactors and cured tuberculosis patients (76.9% versus 40.0%). Conclusion: The increasing expression of MCP-1 mRNA in response to aqueous TSP antigen might be predicted to favor Th1 responses and restricted Th1 responses in treatment failure of pulmonary tuberculosis.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.2
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• pp.125-134
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• 2010

Objective: The aim of this study was to measure the concentrations of cytokines contained in the hydrosalpingeal fluid and to evaluate the effect on the mouse embryo development with the different cytokine concentration. Methods: The hydrosalpingeal fluids (HSF) were collected during the surgery for hydrosalpinx which was confirmed by hysterosalphingogram. The cytokines in HSF including interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-$\alpha$, Interferon-$\gamma$ (IFN-$\gamma$), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), monocyte chemotactic protein-1 (MCP-1) were measured by ELISA method. HSF were added up to culture media with 5%, 10%, and 30% concentrations. The blastulation rates were compared. Results: IL-$\alpha$, IL-$1{\beta}$, IL-2, IL-4, IL-6, IL-8, IL-10, TNF-$\alpha$, IFN-$\bamma$, VEGF, EGF, and MCP-1 were detected, but the concentrations were different from each sample. IL-6 and IL-10 were increased in HSF-1 group, and IFN-$\gamma$, MCP-1, VEGF were increased in HSF-2 compared with normal serum range. The Th1/Th2 ratio of HSF-2 (IFN-$\gamma$:IL10) was highly elevated to 61.64, compared with that of HSF-1 (3.69). The blastulation rate was significantly decreased in HSF-2 group (27.7%) compared those of the HSF-1 group (74%) and control group (76.7%). It showed the trend that the blastulation rate was decreased depending on the concentration HSF of culture media in HSF-2 group, but it was not statistically significant. Conclusion: The composition and concentration of cytokines in each HSF were different, and increased proinflammatory cytokines in HSF might be associated with poor embryonic development. Further study will be needed about the effect of each cytokines in HSF.

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.3
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• pp.276-283
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• 2007

This study was carried out to investigate the effect of isoflavone on the atherogenic effect in C57BL/6 mice. C57BL/6 female mice, 5 weeks of age, were fed on chow diets for 2 weeks during adjustment period. Mice weighing approximately $17.9{\pm}0.9\;g$ were divided into 4 groups and were fed on the experimental diets containing isoflavone for 8 weeks. Experimental groups were control (atherogenic diet), IF-10 (atherogenic diet with isoflavone 10 mg/100 g diet), IF-40 (atherogenic diet with isoflavone 40 mg/100 g diet) and IF-100 (atherogenic diet with isoflavone 100 mg/100 g diet). Food efficiency ratio was not different among the experimental groups. Plasma triglyceride (TG) concentrations were lower after 4 weeks in isoflavone supplementation groups than in control group, whereas monocyte chemoattractant protein-1 and thiobarbituric acid reactive substances (TBARS) levels of plasma were significantly (p<0.05) decreased in the isoflavone supplementation groups in a dose dependent manner. Both hepatic TG and cholesterol levels were significantly lowered in IF-100 than control. Hepatic glutathione concentrations were higher in the IF-100 group than in the other groups. Hepatic antioxidant enzyme activities including glutathione-reductase, glutathione-peroxidase, catalase, and Mn-superoxide dismutase were significantly higher in the isoflavone supplemen-tation groups in a dose dependent manner. From the above results, it is concluded that isoflavone may reduce the risk of atherosclerosis via hypolipidemic, anti-oxidative and anti-inflammatory effects.

• Korean Journal of Poultry Science
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• v.30 no.1
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• pp.67-72
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• 2003

This study was conducted to investigate the effect of germanium biotite on the growth performance, blood composition , fecal NH$_3$-N and volatile fatty acid(VFA) in broiler chicks. Two hundred forty broiler chicks were randomly allocated into five treatments with four replicates for five weeks. Dietary treatments included 1) Control, 2) GB1(basal diet + 200 mesh 0.5%), 3) GB2(basal diet + 325 mesh 0.5%), 4) GB3(basal diet + 200 mesh 1.0%) and 5) GB4(basal diet +325 nesh 1.0%) For overall period, weight gain, feed intake and feed conversion were not significantly different among treatments (P>0.05). Digestibilities of nutrients were not affected by adding GB(P>0.05). No differences occurred in red blood cell(RBC), white blood cell(WBC), hematocrit(HCT), hemoglobin(Hb), lymphocyte, monocyte and platelet of blood among the treatments (P>0.05). Fecal NH$_3$-N concentration of chicks fed GB1.0 diets(GB3 and GB4) was lower(P<0.03) than chicks fed GB0.5 diets(GB1 and GB2). Propionic acid and butyric acid in feces of chicks fed GB diets were lower(P<0.04) than in chicks fed control diet. GB diets reduced fecal acetic acid significantly compared to control (P<0.01). Also, acetic acid of feces in chicks fed GB1.0 diets(GB3 and GB4) was lower(P<0.02) than GB 0.5 diets(GB1 and GB2) treatments. In conclusion, dietary germanium biotite was an effective means of decreasing fecal NH$_3$-N and volatile fatty acid(VFA) emission.

• Korean Journal of Veterinary Research
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• v.28 no.2
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• pp.285-297
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• 1988

The present study was conducted in order to get the normal hematologic values of Korean Jindo dogs. Blood samples were taken from 328(♂ 92, ♂ 236) healthy Jindo dogs in Jindo area. The results obtained in this study were summarized as follows: 1. The mean values of red blood cell(RBC) count, hemoglobin(Hb) content and packed cell volume(PCV) in the age group of less than one year were 6.77(♂ 6.80, ♂ 6.76)${\times}10^6/{\mu}l$, 13.14 (♂ 13.53, ♂ 12.99)g/100ml and 43.28(♂ 44.47, ♂ 42.79)ml/100ml, respectively, whereas the RBC count, Hb content and PCV in the age group of one year and more were 7.42(♂ 7.44, ♂ 7.42)${\times}10^6/{\mu}l$, 14.98(♂ 15.56, ♂ 14.76)g/100ml and 47.18 (♂ 48.43, ♂ 46.71)ml/100ml, respectively. The values of RBC count, Hb content and PCV appeared to be higher in the male than in the female and it increased with aging in the age group of less than one year. 2. The mean values of mean corpuscular volume(MCV), mean corpuscular hemoglobin(MCH) and mean corpuscular hemoglobin content(MCHC) in the age group of less than one year were 63.93(♂ 65.40, ♂ 63.30)fl, 19.41(♂ 19.90, ♂ 19.22) pg and 30.36(♂ 30.43, ♂ 30.36)g/100ml respectively, whereas the MCV, MCH and MCHC in the age group of one year and more were 63.58(♂ 65.09, ♂ 62.95)fl, 20. 19(♂ 20.91, ♂ 19.89) pg and 31.75(♂ 32.13, ♂ 31.60)g/100ml respectively. The values of MCV, MCH and MCHC appeared to be higher in the male than in the female. No differences were found in the MCV and MCH between age groups. The MCHC appeared to be higher in older age group. 3. The mean values of white blood cell(WBC) count in the age group of less than one year were 14,356(♂ 13,878, ♂ 14,551)/${\mu}l$, whereas the values of WBC count in the age group of one year and more were 13,394(♂ 12,656, ♂ 13,672)/${\mu}l$. The WBC count appeared to be higher in the female than in the male. No differences were found between age groups in the WBC count. 4. In WBC differential count the mean percentage of lymphocyte, monocyte, neutrophil and eosinophil of the age group less than one year were 33.45(♂ 31. 64, ♂ 34.18), 2.77(♂ 3.00, ♂ 2.68), 57.40(♂ 58.18, ♂ 57.08) and 6.38(♂ 7.18, ♂ 6.06)% respectively, while those of the age group one year and more were 30. 22(♂ 28.38, ♂ 30.90), 3.03(♂ 3.23, ♂ 2.95), 60.93(♂ 63.34, ♂ 60.03) and 5.82(♂ 5.04, ♂ 6.11)% respectively. Lymphocyte count appeared to be higher in the female and in the age group below one year than in the male and in the age group of one year and more, whereas the reverse was the case with neutrophil and monocyte counts. 5. Mean platelet count in the age group of less than one year was 377,391(♂ 398,778, ♂ 368,721)/${\mu}l$, whereas mean platelet count in the age group of one year and more was 354,657 (♂ 373,660, ♂ 347,512)/${\mu}l$. The platelet count appeared to be higher in the male than in the female and especially lower in the age group of three years and more($305,513/{\mu}l$) than in the other age groups.

• Tuberculosis and Respiratory Diseases
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• v.39 no.4
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• pp.334-342
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• 1992

Background: The activated T lymphocyte by inhalaed mycobacterial antigen may evoke cell-mediated immunity in patients with active pulmonary tuberculosis. These activated lymphocyte may influence the response of tuberculin-purified protein derivative (PPD) in skin test. But occasionally, anergy to PPD appear in patients with pulmonary tuberculosis in spite of active stage. Thus we evaluated the effect of change of subtypes of lymphocyte in bronchoalveolar lavage fluid (BAL) and peripheral blood on anergy to PPD in patients with active pulmonary tuberculosis. Method: We performed tuberculin skin test and flow-cytometry analysis of lymphocytes obtained from BAL fluid and peripheral blood in 11 healthy normal volunteers and 20 patients with active pulmonary tuberculosis. Results: 1) The composition of lymphocyte significantly increased in patients with active pulmonary tuberculosis when compared with that in healthy control ($25.2{\pm}4.8$ vs $6.5{\pm}1.3%$, p<0.01), but composition of monocyte significantly decreased ($69.6{\pm}5.7$ vs $89.2{\pm}1.4%$, p<0.05) in analysis of BAL fluid. 2) There were no differences in compositions of cells in BAL fluid between responders and no-responders to PPD. 3) The compositions of CD3 (+), CD4 (+), CD3 (+) IL-2R (+), CD3 (+) HLA-DR (+) significantly increased in BAL fluid when compared with those in peripheral blood in patients with active pulmonary tuberculosis. But the composition of CDS (+), CD4/CDS were not different between BAL fluid and peripheral blood. 4) There were no correlations between response to PPD and compositions of cells and lymphocyte subtypes in BAL fluid and peripheral blood in all patients with tuberculosis, responders, and no-responders, respectively. Conclusion: From these results, we suggest no direct relationship between compositions of inflammatory cells in bronchoalveolar lavage fluid and we could not rule out the possibility of compartmentalization of activated lymphocyte involving in anergy to PPD in skin test in patients with active pulmonary tuberculosis.

• The Journal of Internal Korean Medicine
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• v.29 no.2
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• pp.334-347
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• 2008

Background and Objective : Haeyeol-tang, composed of Houttuyniae Herba, Lonicerae Flos, Taraxaci Herba, and Scrophulariae Radix, is widely used for alleviating the symptom of various kinds of inflammatory pulmonary disease, including asthma and COPD. We want to know whether Haeyeol-tang has an anti-inflammatory effect by analyzing expression of pro-inflammatory cytokines. Materials and Methods : We differentiated the THP-1 cells into macrophage-like cells by treatment with PMA. Inflammation was induced by treatment with LPS and PMA. We found the safe concentration of Haeyeol-tang by using MTS assay and used PD98059 as a negative control for comparison of anti-inflammatory effect of Haeyeol-tang. Results : The RT-PCR analysis results show that the cell survival rate is over 100% within 1 ng/mL to 1 ug/mL of Haeyeol-tang and begins to decrease under 100% at 10 ug/mL. The gene expression of $IL-1{\beta}$, IL-6, IL-8, IL-10, $TNF-{\alpha}$ and $TGF-{\beta}$ levels were down-regulated when Haeyeol-tang was treated at concentrations between 1 ng/mL an 1 ug/mL on monocyte-derived macrophages. Interestingly, 1 ug/mL Haeyeol-tang-treated samples showed that the transcriptional activities of IL-8, $TNF-{\alpha}$, IL-10 and $TGF-{\beta}$ were more down-regulated than those of PD098059 $(TNF-{\alpha}$ inhibitor). At protein level, the ELISA analysis results showed that there were more remarkable (p<0.001) decreases in expression of $IL-1{\beta}$, IL-6, IL-8 and $TNF-{\alpha}$ on both the 1 ug/mL Haeyeol-tang-treated group and the PD98059-treated group than the LPS-treated group. Conclusion : We conclude that Haeyeol-tang has an anti-inflammatory effect by inhibiting expression of pro-inflammatory cytokines and chemokines at mRNA and protein levels. These results may provide us a promising way to care for general inflammatory diseases as well as inflammatory pulmonary disease, including asthma and COPD, with further clinical study.

• Korean Journal of Veterinary Research
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• v.51 no.4
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• pp.259-265
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• 2011

Several compounds and extracts isolated from a brown alga, Ishige (I.) okamurae, exhibit anti-oxidant and anti-inflammatory effects. The present study investigated whether the ethyl acetate (EtOAc) fraction of I. okamurae (EFIO) could ameliorate carbon tetrachloride ($CCl_{4}$)-induced hepatotoxicity in rats. Sprague-Dawley rats were intraperitoneally (i.p.) administered with EFIO at 10 or 50 mg/kg per day for 2 consecutive days before $CCl_{4}$ injection (3.3 mL/kg, i.p.). Twenty four hours later, the rats were anesthesized with diethyl ether and dissected. Pretreatment with EFIO significantly reduced the increased serum levels of alanine aminotransferase and aspartate aminotransferase in $CCl_{4}$-treated rats. Pretreatment with EFIO also significantly inhibited the reduced activities of superoxide dismutase and catalase in the $CCl_{4}$-injured liver. Histopathological evaluations showed that hemorrhage, hepatocyte necrosis, inflammatory cell infiltration, and fatty degeneration induced by $CCl_{4}$ treatment were ameliorated by the administration of EFIO. Additionally, liver immunohistochemical analyses revealed the marked reduction in ED1-positive monocyte-like macrophages in EFIO-pretreated rats given $CCl_{4}$. These results suggest that EFIO ameliorates $CCl_{4}$-induced liver injury, possibly through the inhibition of oxidative stress.