Spinal tuberculosis (ST) is the tuberculosis caused by Mycobacterium tuberculosis (Mtb) infections in spinal curds. Isoliquiritigenin (4,2',4'-trihydroxychalcone, ISL) is an anti-inflammatory flavonoid derived from licorice (Glycyrrhiza uralensis), a Chinese traditional medicine. In this study, we evaluated the potential of ISL in treating ST in New Zealand white rabbit models. In the model, rabbits (n=40) were infected with Mtb strain H37Rv or not in their $6^{th}$ lumbar vertebral bodies. Since the day of infection, rabbits were treated with 20 mg/kg and 100 mg/kg of ISL respectively. After 10 weeks of treatments, the adjacent vertebral bone tissues of rabbits were analyzed through Hematoxylin-Eosin staining. The relative expression of Monocyte chemoattractant protein-1 (MCP-1/CCL2), transcription factor ${\kappa}B$ ($NF-{\kappa}B$) p65 in lymphocytes were verified through reverse transcription quantitative real-time PCR (RT-qPCR), western blotting and enzyme-linked immunosorbent assays (ELISA). The serum level of interleukin (IL)-2, IL-4, IL-10 and interferon ${\gamma}$ ($IFN-{\gamma}$) were evaluated through ELISA. The effects of ISL on the phosphorylation of $I{\kappa}B{\alpha}$, $IKK{\alpha}/{\beta}$ and p65 in $NF-{\kappa}B$ signaling pathways were assessed through western blotting. In the results, ISL has been shown to effectively attenuate the granulation inside adjacent vertebral tissues. The relative level of MCP-1, p65 and IL-4 and IL-10 were retrieved. $NF-{\kappa}B$ signaling was inhibited, in which the phosphorylation of p65, $I{\kappa}B{\alpha}$ and $IKK{\alpha}/{\beta}$ were suppressed whereas the level of $I{\kappa}B{\alpha}$ were elevated. In conclusion, ISL might be an effective drug that inhibited the formation of granulomas through downregulating MCP-1, $NF-{\kappa}B$, IL-4 and IL-10 in treating ST.
Flammulina velutipes is an edible mushroom and contains a lot of fiber, vitamin $B_1$, $B_2$, niacin and folic acid. This study was conducted to explore the effects of the Flammulina velutipes mushroom on immune cells and immunity. Th1 cytokine productions as $IFN-{\gamma}$, $TNF-{\alpha}$, and IL-2 were measured in an activated macrophage by Flammulina velutipes water extract in seven concentrations (0, 5, 10, 50, 100, 250, 500, and $1,000{\mu}g/mL$). Also, the splenocyte proliferation index was measured at 48 hours after treatment of the Flammulina velutipes water extract in seven concentrations or mitogen, LPS and ConA. The $IFN-{\gamma}$ and $TNF-{\alpha}$ productions were increased by treatment of the Flammulina velutipes water extract. The $TNF-{\alpha}$ production was significantly higher in the $50{\sim}1,000{\mu}g/mL$ Flammulina velutipes water extract treated macrophages. The $IFN-{\gamma}$ production of macrophages treated with the Flammulina velutipes water extract increased significantly in all groups, and the highest $1000{\mu}g/mL$ concentration. The splenocyte proliferation index was enhanced when the $10{\sim}1,000{\mu}g/mL$ Flammulina velutipes water extracts were treated compared to the control. These primary results suggest that Flammulina velutipes may enhance the immune function by activation of the macrophage and spleen cell.
Guo, Jian-Rong;Xu, Feng;Jin, Xiao-Ju;Shen, Hua-Chun;Liu, Yang;Zhang, Yi-Wei;Shao, Yi
Asian Pacific Journal of Cancer Prevention
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v.15
no.1
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pp.467-474
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2014
Objective: To observe the effects of allogeneic and autologous transfusion on cellular immunity, humoral immunity and secretion of serum inflammatory factors and perforin during the perioperative period in patients with malignant tumors. Methods: A total of 80 patients (age: 38-69 years; body weight: 40-78 kg; ASA I - II) receiving radical operation for gastro-intestinal cancer under general anesthesia were selected. All the patients were divided into four groups based on the methods of infusion and blood transfusion: blank control group (Group C), allogeneic transfusion group (group A), hemodiluted autotransfusion Group (Group H) and hemodiluted autotransfusion + allogenic transfusion Group (A+H group). Venous blood was collected when entering into the surgery room ($T_0$), immediately after surgery ($T_1$) and 24h ($T_2$), 3d ($T_3$) and 7d ($T_4$) after surgery, respectively. Moreover, flow cytometry was applied to assess changes of peripheral blood T cell subpopulations and NK cells. Enzyme linked immunosorbent assays were performed to determine levels of IL-2, IL-10, TNF-${\alpha}$ and perforin. Immune turbidimetry was employed to determine the changes in serum immunoglobulin. Results: Both CD3+ and NK cells showed a decrease at $T_1$ and $T_2$ in each group, among which, in group A, CD3+ decreased significantly at $T_2$ (P<0.05) compared with other groups, and CD3+ and NK cell reduced obviously only in group A at $T_3$ and $T_4$ (P<0.05). CD4+ cells and the ratio of D4+/CD8+ were decreased in groups A, C and A+H at $T_1$ and $T_2$ (P<0.05). No significant intra- and inter-group differences were observed in CD8+ of the four groups (P<0.05). IL-2 declined in group C at $T_1$ and $T_2$ (P<0.05) and showed a decrease in group A at each time point (P<0.05). Moreover, IL-2 decreased in group A + H only at $T_1$. No significant difference was found in each group at $T_1$ (P<0.05). More significant decrease in group ?? at $T_2$, $T_3$ and $T_4$ compared with group A (P<0.05), and there were no significant differences among other groups (P>0.05). IL-10 increased at $T_1$ and $T_2$ in each group (P<0.05), in which it had an obvious increase in group A, and increase of IL-10 occurred only in group A at $T_3$ and $T_4$ (P<0.05). TNF-${\alpha}$ level rose at $T_1$ (P<0.05), no inter- and intra-group difference was found in perforin in all groups (P<0.05). Compared with the preoperation, both IgG and IgA level decreased at $T_1$ in each group (P<0.05), and they declined only in Group A at $T_2$ and $T_3$ (P<0.05), and these parameters were back to the preoperative levels in other groups. No significant differences were observed between preoperative and postoperative IgG and IgA levels in each group at $T_4$ (P>0.05). No obvious inter- and intra-group changes were found in IgM in the four groups (P>0.05). Conclusions: Allogeneic transfusion during the perioperative period could obviously decrease the number of T cell subpopulations and NK cells and the secretion of stimulating cytokines and increase the secretion of inhibiting cytokines in patients with malignant tumors, thus causing a Th1/Th2 imbalance and transient decreasing in the content of plasma immune globulin. Autologous transfusion has little impact and may even bring about some improvement oo postoperative immune function in patients with tumors. Therefore, cancer patients should receive active autologous transfusion during the perioperative period in place of allogeneic transfusion.
Purpose : This study was performed to evaluate anti-thrombotic and anti-inflammatory effects of BokbangHongdeungPaejangSan water extract (BHPS). Methods : BHPS was investigated using cultured cells and a murine models. As for the parameters of inflammation, levels of several inflammatory cytokines and chemical mediators which are known to be related to inflammation were determined in mouse lung fibroblast cells (mLFC) and RAW 264.7 cells. Results : In experiment of anti-thrombotic effect, BHPS inhibited the platelet aggregation induced by ADP and epinephrine, and inhibited pulmonary embolism induced by collagen and epinephrine. BHPS increased Platelet number and fibrinogen amount, and shortened PT and APTT in thrombus model induced by dextran. In experiment of anti-inflammatory effect, BHPS inhibited IL-1${\beta}$, IL-6, TNF-${\alpha}$, COX-2 and NOS-II mRNA expression in a concentration-dependent manner in RAW 264.7 cell line, and inhibited significantly NO production at 50, 100 ${\mu}g/ml$, and also inhibited ROS production in a concentration-dependent manner. BHPS inhibited IL-1${\beta}$, IL-6 and TNF-${\alpha}$ production significantly in serum of acute inflammation-induced Balb/c mice, and decreased IL-1${\beta}$, IL-6 and TNF-${\alpha}$ production in spleen tissue, but increased IL-1${\beta}$, IL-6 and TNF-${\alpha}$ production in liver tissue. BHPS increased survival rate at the 3th day in ICR mice with lethal endotoxemia induced by LPS. Conclusion : These results suggest that BHPS can be used for treating diverse female diseases caused by thrombosis and inflammation such as endometriosis, pelvic pain, cervicitis, pelvic inflammatory disease and pelvic tuberculosis and so forth.
Purpose: This study was performed to evaluate anti-thrombotic and anti-inflammatory effects of NeungaSoJeokTang water extract (NSJT). Methods: In the study of anti-inflammatory effects, NSJT was investigated using cultured cells and murine models. As for the parameters of inflammation, levels of several inflammatory cytokines and chemical mediators which are known to be related to inflammation were determined in mouse lung fibroblast cells(mLFC) and RAW 264.7 cells. Results: Prior to the experiment, we evaluated sGOT, sGPT, BUN and creatine after the treatment. As a result, NSJT was innoxious on liver and kidney. In experiment of anti-thrombotic effect, NSJT inhibited the platelet aggregation induced by ADP and epinephrine, and inhibited pulmonary embolism induced by collagen and epinephrine. NSJT did not affect significantly the blood flow rate both in vitro and in vivo. NSJT increased platelet number and fibrinogen amount, and NSJT shortened PT and APTT in thrombus model induced by dextran. In experiment of anti-inflammatory effect, NSJT inhibited $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, COX-2 and NOS-II mRNA expression in a concentration-dependent manner in RAW 264.7 cell line, and inhibited significantly NO production at 50, 100 ${\mu}g/ml$, and also inhibited ROS production in a concentration-dependent manner. NSJT inhibited $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production significantly in serum of acute inflammation-induced Balb/c mice, and decreased $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production in spleen tissue, but increased $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ production in liver tissue. NSJT increased survival rate at the 3th day in ICR mice with lethal endotoxemia induced by LPS. Conclusion: These results suggest that NSJT can be used for treating diverse female diseases caused by thrombosis and inflammation such as pelvic pain, pelvic inflammatory disease as well as vulvar pain due to vulvitis, vulvar vestibulitis and so on.
Yoon, Hyun-A;Han, Young-Woo;Aleyas, Abi George;George, June Abi;Kim, Seon-Ju;Kim, Hye-Kyung;Song, Hee-Jong;Cho, Jeong-Gon;Eo, Seong-Kug
Journal of Microbiology and Biotechnology
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v.18
no.3
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pp.591-599
/
2008
A murine model immunized by systemic and mucosal delivery of plasmid DNA vaccine expressing glycoprotein B (pCIgB) of pseudorabies virus (PrV) was used to evaluate both the nature of the induced immunity and protection against a virulent virus. With regard to systemic delivery, the intramuscular (i.m.) immunization with pCIgB induced strong PrV-specific IgG responses in serum but was inefficient in generating a mucosal IgA response. Mucosal delivery through intranasal (i.n.) immunization of pCIgB induced both systemic and mucosal immunity at the distal mucosal site. However, the levels of systemic immunity induced by i.n. immunization were less than those induced by i.m. immunization. Moreover, i.n. genetic transfer of pCIgB appeared to induce Th2-biased immunity compared with systemic delivery, as judged by the ratio of PrV-specific IgG isotypes and Th1- and Th2-type cytokines produced by stimulated T cells. Moreover, the immunity induced by i.n. immunization did not provide effective protection against i.n. challenge of a virulent PrV strain, whereas i.m. immunization produced resistance to viral infection. Therefore, although i.n. immunization was a useful route for inducing mucosal immunity at the virus entry site, i.n. immunization did not provide effective protection against the lethal infection of PrV.
Park, Yeong-Chul;Lim, Jung-Dae;Park, Yong-Ki;Yoon, Mi-Sook;Lee, Sun-Dong
The Korea Journal of Herbology
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v.27
no.4
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pp.33-44
/
2012
Objectives : There is a pressing need to determine the clinical and scientific validity of herbal therapies for animal model with atopic dermatitis since some differences in systemic cytokine polarization between in animal model and in patients with atopic dermatitis has been reported. New studies for tang, medicinal herb itself or effective ingradients of medicinal herb showing anti-atopic dermatitis effectiveness are reviewed in terms of cytokine regulation. Methods : Those herbal therapies used to treat atopic dermatitis in animal model were introduced and the expression pattern of cytokine and the activity of mast cell were compared in both animal model and patients with atopic dermatitis. Results : In case of atopic dermatitis in human, there is a biphasic pattern of cytokine expression in atopic dermatitis, with acute skin inflammation associated with a predominance of IL-4 and IL-13 expression from Th2 cells, and chronic inflammation associated with increased IL-5 from Th2-cells and IFN-${\gamma}$ from Th1-cells. However, a pattern of cytokine expression in animal model with atopic dermatitis is not matched well to the biphasic pattern of cytokine expression in patients with atopic dermatitis. In addition, a kind of cytokine is different by animal model with atopic dermatitis. These differences would make herbal medicines, showing their effectiveness on atopic dermatitis, difficult to apply to patients with atopic dermatitis. Conclusion : The pattern of local cytokine expression plays an important role in modulating tissue inflammation, and in atopic dermatitis this pattern depends on the acuity or duration of the skin lesion. Thus, in order to develop medicinal herb itself or effective ingradients of medicinal herb showing anti-atopic dermatitis effectiveness, biphasic pattern of cytokine expression should be considered in animal model with atopic dermatitis.
Doroudchi, Mehrnoosh;Pishe, Zahra Ghanaat;Malekzadeh, Mahyar;Golmoghaddam, Hossein;Taghipour, Mousa;Ghaderi, Abbas
Asian Pacific Journal of Cancer Prevention
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v.14
no.9
/
pp.5225-5230
/
2013
Background: There is a Th1/Th2 cytokine imbalance and expression of IL-17 in patients with brain tumours. We aimed to compare the levels of IL-17A and IL-6 in sera of glioma, meningioma and schwannoma patients as well as in healthy individuals. Materials and Methods: IL-17A and IL-6 levels were measured in sera of 38 glioma, 24 meningioma and 18 schwannoma patients for comparison with 26 healthy controls by commercial ELISA assays. Results: We observed an increase in the IL-17A in 30% of glioma patients while only 4% and 5.5% of meningioma and schwannoma patients and none of the healthy controls showed elevated IL-17A in their sera ($0.29{\pm}0.54$, $0.03{\pm}0.15$ and $0.16{\pm}0.68$ vs. $0.00{\pm}0.00pg/ml$; p=0.01, p=0.01 and p=0.001, respectively). There was also a significant decrease in the level of IL-6 in glioma patients compared to healthy controls ($2.34{\pm}4.35$ vs. $4.67{\pm}4.32pg/ml$; p=0.01). There was a direct correlation between the level of IL-17A and age in glioma patients (p=0.005). Glioma patients over 30 years of age had higher IL-17A and lower IL-6 in their sera compared to the young patients. In addition, a non-significant grade-specific inverse trend between IL-17A and IL-6 was observed in glioma patients, where high-grade gliomas had higher IL-17A and lower IL-6. Conclusions: Our data suggest a Th17 mediated inflammatory response in the pathogenesis of glioma. Moreover, tuning of IL-6 and IL-17A inflammatory cytokines occurs during progression of glioma. IL-17A may be a potential biomarker and/or immunotherapeutic target in glioma cases.
2Department of Marine Molecular Biotechnology, College of Life Science, Gangneung-Wonju National University Recently, we reported that diarylheptanoid hirsutenone (HST) effectively inactivated T lymphocytes. However, it has not been evaluated whether HST is involved in calcineurin or calmodulin inactivation. In the present study, cells were treated with T-cell inhibitors with or without HST. Our results revealed that HST successfully inhibited expression of T-helper type I (Th1) and Th2 cytokines. Co-treatment with HST and nuclear factor-activated T cell (NFAT) activation inhibitor III (INCA-6) showed a more sensitive effect than that with other inhibitors, suggesting that HST contributes to inhibition of dephosphorylation of NFAT in the cytosol. HST up-regulated cell cycle arrest genes and inhibited the growth of Staphylococcus aureus. These effects were confirmed in an NFAT electrophoretic-mobility shift assay via successful inhibition of NFAT translocation and in the histological recovery in a 2,4-dinitrochloro benzene-induced in vivo model. Taken together, HST was shown to effectively inhibit T-cell activation via inhibition of cytosolic NFAT dephosphorylation, similar to INCA-6.
Hand-foot-and-mouth disease (HFMD) is a viral infectious disease that occurs in children under 5 years of age. Its main causes are coxsackievirus (CV) and enterovirus (EV). Since there are no efficient therapeutics for HFMD, vaccines are effective in preventing the disease. To develop broad coverage against CV and EV, the development of a bivalent vaccine form is needed. The Mongolian gerbil is an efficient and suitable animal model of EV71 C4a and CVA16 infection used to investigate vaccine efficacy following direct immunization. In this study, Mongolian gerbils were immunized with a bivalent inactivated EV71 C4a and inactivated CVA16 vaccine to test their effectiveness against viral infection. Bivalent vaccine immunization resulted in increased Ag-specific IgG antibody production; specifically, EV71 C4a-specific IgG was increased with medium and high doses and CVA16-specific IgG was increased with all doses of immunization. When gene expression of T cell-biased cytokines was analysed, Th1, Th2, and Th17 responses were found to be highly activated in the high-dose immunization group. Moreover, bivalent vaccine immunization mitigated paralytic signs and increased the survival rate following lethal viral challenges. When the viral RNA content was determined from various organs, all three doses of bivalent vaccine immunization were found to significantly decrease viral amplification. Upon histologic examination, EV71 C4a and CVA16 induced tissue damage to the heart and muscle. However, bivalent vaccine immunization alleviated this in a dose-dependent manner. These results suggest that the bivalent inactivated EV71 C4a/CVA16 vaccine could be a safe and effective candidate HFMD vaccine.
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