• 대한한의학회지
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• 제24권2호
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• pp.148-158
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• 2003

Backgrounds & Objectives: In many recent studies, molecular biological methods have been used to investigate the role of cytokines in pathogenesis and new therapeutic targets of asthma. Recently, as a method of research on the gene expression, they are applying another method which assays multiple gene expressions at the same time by the microarray. In this study, the antioxidant effect, the inhibitory effect against interleukin-4 and the effect on the CD/cytokine gene expression in PBMC (peripheral blood mononuclear cells) was evaluated by using cDNA microarray chip of Chungsangboha-tang. Methods: Experimental studies were performed for the antioxidant effect of Chungsangboha-tang on DPPH (1, 1-diphenyl-2-picrylhydrazyl) solution, for the IL-4-inhibiting effect on BALB/c mouse spleen, and for the gene expression effect on PBMC (peripheral blood mononuclear cells) with microarray. Results: Chungsangboha-tang showed antioxidant effect dose-dependently. Chungsangboha-tang inhibited interleukin-4 dose-dependently and showed significant difference in 10ug/ml and 100ug/ml of test groups. There was no 2 more times upregulated genes than in the control group by using cDNA microarray chip of Chungsangbohn-tang, but there were 140%-200% upregulated genes. There was no 2 more times downregulated genes than in the control group by using cDNA microarray chip of Chungsangboha-Tang, but there was 50%-75% downregulated genes. Conclusions: This study showed that Chungsangboha-tang has an antioxidant effect and inhibition of Interleukin-4, but further studies are necessary with microarray.

• Asian Pacific Journal of Cancer Prevention
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• 제15권1호
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• pp.265-271
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• 2014

Interleukin-7 (IL-7) is a potent anti-apoptotic cytokine that enhances immune effector cell functions and is essential for lymphocyte survival. While it known to induce differentiation and proliferation in some haematological malignancies, including certain types of leukaemias and lymphomas, little is known about its role in solid tumours, including breast cancer. In the current study, we investigated whether IL-7 could enhance the in vivo antitumor activity of tumor-reactive $CD8^+$ T cells with induction of IFN-${\gamma}$ in a murine breast cancer model. Human IL-7 cDNA was constructed into the eukaryotic expression plasmid pcDNA3.1, and then the recombinational pcDNA3.1-IL-7 was intratumorally injected in the TM40D BALB/C mouse graft model. Serum and intracellular IFN-${\gamma}$ levels were measured by ELISA and flow cytometry, respectively. $CD8^+$ T cell-mediated cytotoxicity was analyzed using the MTT method. Our results showed that IL-7 administration significantly inhibited tumor growth from day 15 after direct intratumoral injection of pcDNA3.1-IL-7. The anti-tumor effect correlated with a marked increase in the level of IFN-${\gamma}$ and breast cancer cells-specific CTL cytotoxicity. In vitro cytotoxicity assays showed that IL-7-treatment could augment cytolytic activity of $CD8^+$ T cells from tumor bearing mice, while anti-IFN-${\gamma}$ blocked the function of $CD8^+$ T cells, suggesting that IFN-${\gamma}$ mediated the cytolytic activity of $CD8^+$ T cells. Furthermore, in vivo neutralization of $CD8^+$ T lymphocytes by CD8 antibodies reversed the antitumor benefit of IL-7. Thus, we demonstrated that IL-7 exerts anti-tumor activity mainly through activating $CD8^+$ T cells and stimulating them to secrete IFN-${\gamma}$ in a murine breast tumor model. Based on these results, our study points to a potential novel way to treat breast cancer and may have important implications for clinical immunotherapy.

• Asian-Australasian Journal of Animal Sciences
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• 제30권7호
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• pp.912-919
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• 2017

Objective: Identification of the candidate genes that play key roles in phenotypic variations can provide new information about evolution and positive selection. Interleukin (IL)-32 is involved in many biological processes, however, its role for the immune response against various diseases in mammals is poorly understood. Therefore, the current investigation was performed for the better understanding of the molecular evolution and the positive selection of single nucleotide polymorphisms in IL-32 gene. Methods: By using fixation index ($F_{ST}$) based method, IL-32 (9375) gene was found to be outlier and under significant positive selection with the provisional combined allocation of mean heterozygosity and $F_{ST}$. Using nucleotide sequences of 11 mammalian species from National Center for Biotechnology Information database, the evolutionary selection of IL-32 gene was determined using Maximum likelihood model method, through four models (M1a, M2a, M7, and M8) in Codeml program of phylogenetic analysis by maximum liklihood. Results: IL-32 is detected under positive selection using the $F_{ST}$ simulations method. The phylogenetic tree revealed that goat IL-32 was in close resemblance with sheep IL-32. The coding nucleotide sequences were compared among 11 species and it was found that the goat IL-32 gene shared identity with sheep (96.54%), bison (91.97%), camel (58.39%), cat (56.59%), buffalo (56.50%), human (56.13%), dog (50.97%), horse (54.04%), and rabbit (53.41%) respectively. Conclusion: This study provides evidence for IL-32 gene as under significant positive selection in goat.

• The Korean Journal of Physiology
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• 제29권1호
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• pp.1-11
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• 1995

Alveolar macrophages play a pivotal role in the pathogenesis of silicosis since the macrophages may release a wide variety of toxic and inflammatory mediators as well as mitogenic growth factors. In the present study, the effects of in vitro exposure to silica on release of various mediator such as reactive oxygen species, platelet activating factor(PAF), and interleukin-1 (IL-1) by alveolar macrophages were examined. First, hydrogen peroxide release from alveolar macrophages was monitored by measuring the change in fluorescence of scopoletin in the absence or presence of graded concentration of silica. Significantly enhanced release of hydrogen peroxide was observed at 0.5 mg/ml and above. A maximal enhancement of 10 fold above control was observed at 5 mg/ml silica. Similarly, in vitro exposure to silica also significantly stimulated the generation of chemiluminescence from alveolar macrophages at 0.5 mg/ml and above with n maximal enhancement of 8 fold at 5 mg/ml silica. Second, PAF release from alveolar macrophages after 30 min incubation at $37^{\circ}C$ in absence or presence of zymosan and silica was determined by measuring $^{3}H-serotonin$ release ability of the conditioned macrophage supernates from platelets. 5 mg/ml zymosan as a positive control fur the PAF assay increased PAF release by 19 % of total serotonin release. Furthermore, silica also resulted in significant enhancement of the PAF release compared with that in unstimulated (control) cells, i.e., $17.7{\pm}5.8%$ and $24.0{\pm}4.9%$ of total serotonin release at 5 mg/ml and 10 mg/ml silica, respectively, which represents the release of nanomole levels of PAF. Lastly, IL-1 production by alveolar macrophages was analysed following their stimulation with lipopolysaccharide (LPS) and silica by their capacity to stimulate thymocyte proliferation. $10\;{\mu}g/ml$ LPS resulted in an 11 fold increase in IL-1 production. In comparison, $50\;{\mu}g/ml$ silica resulted in a 4 fold increase in IL-1 release. These data indicate that in vitro exposure of alveolar macrophages to silica activates the release of various bioactive mediators such as reactive oxygen species, PAF and IL-1 which thus contribute to amplification of inflammatory reactions and regulation of fibrotic responses by the lung after inhalation of silica.

• Journal of the Korean Wood Science and Technology
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• 제41권6호
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• pp.551-558
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• 2013

본 연구는 만병초와 구상나무 추출물의 항알러지 효과에 대해 조사하였다. 비만세포 RBL-2H3에 concanavalin A (Con A)를 처리하여 면역반응을 유도한 후 만병초와 구상나무 추출물을 농도별로 처리하여 real-time PCR을 이용하여 IL-4, IL-13의 mRNA 발현량을 확인하였다. 또한 ${\beta}$-hexosaminidase 분비를 측정하여 탈과립 유도효과를 평가하였다. IL-4의 발현은 만병초 $10^{-7}$, $10^{-5}$, $10^{-3}%$ 농도와 구상나무의 모든 농도에서 유의성 있게 감소하였으며, IL-13의 발현은 만병초와 구상나무 모든 농도에서 유의성 있게 감소되었다. 만병초 추출물과 구상나무 추출물의 $10^{-5}$, $10^{-3}%$ 농도에서는 ${\beta}$-hexosaminidase 분비가 유의성 있게 감소하였다. 이러한 결과를 통해 만병초와 구상나무 추출물이 면역반응 표적유전자의 발현을 감소시키고 면역반응에 의해 유도되는 탈과립유도 효과도 감소시킴을 확인할 수 있었다. 이는 만병초와 구상나무 추출물이 항알러지 효과가 있음을 제시하는 결과이다.

• 한국발생생물학회지:발생과생식
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• 제22권2호
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• pp.155-163
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• 2018

This study aimed to investigate changes in the activity and mRNA expression of plasminogen activators (PAs) induced by $17{\beta}$-estradiol ($E_2$), human chorionic gonadotropin (hCG), and interleukin-$1{\beta}$ ($IL-1{\beta}$) in porcine endometrial cells. Endometrial cells were isolated from the epithelium and cultured to 80% confluence. They were then treated for 24 h with $E_2$ (0.2, 2, 20, and 200 ng/mL), $IL-1{\beta}$ (0.1, 1, 10, and 100 ng/mL), and hCG (0.5, 1, 1.5 and 2 IU/mL). mRNA expressions of urokinase-type (uPA) and tissue-type (tPA) PAs were analyzed using reverse transcription PCR, and activities were measured using a PA activity assay. mRNA expressions of uPA and tPA increased with $E_2$ treatment; however, this was not significant. Similarly, treatment with hCG did not influence the mRNA expressions of PAs. Interestingly, treatment with 0.1 ng/mL $IL-1{\beta}$ significantly reduced the mRNA expression of uPA, but did not affect that of tPA. Treatment with 2, 20, and 200 ng/mL $E_2$ increased PA activity compared with the control group; treatment with 0.1 and 1 ng/mL $IL-1{\beta}$ significantly increased PA activity compared with the other $IL-1{\beta}$ treatment groups, whereas treatment with 10 and 100 ng/mL $IL-1{\beta}$ decreased. Treatment with 2 IU/mL hCG increased PA activity compared with the other treatment groups, although there were no significant differences between the hCG and control groups. In conclusion, the activity and mRNA expression of PAs were differently regulated by the hormone/cytokine and its concentration in porcine endometrial cells. Therefore, understanding PA regulatory mechanisms may help to improve the reproductive potential of domestic animals.

• Restorative Dentistry and Endodontics
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• 제30권1호
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• pp.1-6
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• 2005

근관치료의 실패원인 중 중요한 세균으로 알려진 Enterococcus faecalis는 최근에 중요성이 더해지며 많은 연구들이 진행중이다. 여러가지 기전들이 보고되고 있으나 면역반응에 관한 연구는 거의 알려져 있지 않은 상태이다. 본 연구에서는 Enterococcus faecalis의 초음파 분쇄 추출물을 성인의 말초혈액으로부터 얻은 임파구에 적용시켜서 여기서 분비되는 interleukin-2, interleukin-4, transforming growth $factor-\beta1$의 농도를 Enzyme linked immunosorbent assay (ELISA)로 측정하여 비교, 평가하는 것을 목적으로 한다. E. faecalis를 적절한 조건에서 배양한 뒤 초음파 분쇄를 하여 추출물을 얻어냈다. 임파구는 건강한 성인의 말초혈액에서 추출하여 분리하였다. 임파구를 적절한 농도의 mitogen (Phytohemagglutinin: PHA)으로 자극시킨 뒤에 다양한 농도의 E. faecalis 초음파 추출물을 적용시키고 72시간 동안 배양하였다 ELISA를 이용하여 IL-2, IL-4, $TGF-\beta1$의 농도를 측정하였다. 실험결과는 Kruskal-Wallis test, Man-Whitney rank sum test (p < 0.05)를 사용하여 통계처리 하였다. 실험결과 PHA로 처리한 군은 아무것도 처리하지 않은 군에 비해서 IL-2, IL-4의 수치가 유의성 있게 높았다 (p < 0.05). PHA로 처리한 군중에서 고농도와 중농도의 sonic extract of E. faecalis (SEF)로 처리한 군은 그렇지 않은 군에 비해서 IL-2 IL-4의 농도가 유의성 있게 낮았다 (p < 0.05). PHA로 처리한 군중에서 저농도의 SEF로 처리한 군은 그렇지 않은 군과 비교하여 유의할 만한 차이를 보이지 않았다. $TGF-\beta1$의 농도는 모든 군에서 유의할 만한 차이를 보이지 않았다 (p > 0.05). 따라서, E. faecalis의 추출물은 임파구의 IL-2, IL-4의 분비능력을 저하시킨다고 할 수 있다.

• Parasites, Hosts and Diseases
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• 제31권3호
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• pp.249-258
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• 1993

원발성 아메바성 수막뇌염을 야기하는 Naegleria fowleri를 사망율에 따라 $1{\;}{\times}{\;}10^4$개 아메바 영양형 접종군과. $1{\;}{\times}{\;}10^5$개 아메바 영양형 접종군으로 나누어 비장세포를 비특이 mitogen인 Phytohemagglutinin(PHA)과 특이항원인 N. fowleri Iysates로 자극하여 T 세포의 interleukin-2 생성정도를 측정하고 T 림프구의 아형 및 아세포화정도를 측정한 결과 N.fowleri 아메바 영양형 $1{\;}{\times}{\;}10^5$개 접종군에서의 마우스 사망율은 72.2%였으며 $1{\;}{\times}{\;}10^4$개 접종군에서의 마우스 사망율은 14.3%였다. 또한 IL릭 생성정도를 접종후 7, 14, 24일째 측정한 결과. 접종 후 14일째에는 두 실험군 모두에서 대조군과 비교하여 IL닉의 생성이 유의하게 감소하였으며 접종 후 24일째에는 두 실험군 모두에서 접종 후 14일째 보다는 증가하였으나 대조군과 비교하여 유의하게 억제되었다 비장세포내 T 림프구 아형의 변동은 전체 비장 림프구에 대한 $Thy-1.2^{+}{\;}T.{\;}L3T4^{+}{\;}T,{\;}Ly^{2+}{\;}T$ 세포는 N.fowleri아메바 영양형 $1{\;}{\times}{\;}10^5$개 접종군에서 접종 후 7일째 대조군과 비교하여 유의하게 증가하였고 접종 후 14일째와 24일째에는 대조군과 비교하여 유의한 차이가 없었다. 아메바 영양형 $1{\;}{\times}{\;}10^4$개를 접종시킨 실험군은 경과 일수에 관계없이 대조군과 비교하여 유의한 차이가 없었다. 비장세포내 T 세포의 DNA의 분획을 관찰한 결과 두 실험군 모두에서 접종 후 7일째에 S phase 분획이 가장 높이 증가하였으며 접종 후 14일과 24일째에도 대조군에 비하여는 유의하게 증가하여 있었다. 이상의 결과를 종합하여 볼 때, N.fowleri를 사망율을 기준으로 접종량을 달리하여 아메바 영양형 $1{\;}{\times}{\;}10^4$개 접종군과 $1{\;}{\times}{\;}10^5$개 접종군으로 나누어 접종하였을 때, 접종 후 7일을 전후하여 IL-2를 매개로 활성화되는 세포매개성 면역이 N.fowleri감염의 방어기작으로 작용하는 것으로 생각되며 아메바 접종 후 14일째에는 치명적인 수막뇌염으로 진행되어 비장세포의 IL-2의 생성능력이 매우 억제된 것으로 생각된다. 또한 IL-2 생성능력과 T 세포의 아세포화의 증가 및 T세포 아령의 수의 변동과는 잘 일치되지 않는 것으로 나타났다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권7호
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• pp.2951-2957
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• 2014

Background: Gastric carcinogenesis is a complicated process that involves environmental and genetic factors like interleukin-4 (IL-4) and IL-8. Single nucleotide polymorphisms in their genes are associated with changed levels of gene expression. Here, we investigated the association between IL4-590 C>T and IL8-251T>A and gastric cancer (GC) risk in Sichuan of Southwestern China. Materials and Methods: We surveyed the research subjects using a self-designed questionnaire with questions on demographic factors and putative risk factors. Approximately 2-5ml of whole blood was collected after field survey to analyze IL4-590 C>T and IL8-251T>A genotypes using MALDI-TOF MS. Results: Our study recruited 308 pairs of GC patients and controls, including 224 (72.7%) men and 84 (27.3%) women in each group. There were 99 cardia and 176 noncardia GC patients in the case group. The case and control groups had an average age of $57.7{\pm}10.6$ ($mean{\pm}SD$) and $57.6{\pm}11.1$ years. GC patients reported a significantly greater proportion of family history of cancer (29.9% vs 10.7%, p<0.01) and drinking (54.6% vs 43.2%, p<0.01) than did controls. Variant genotypes of IL-4-590 C>T and IL-8-251 T>A were not associated with overall GC risk (adjusted OR, 0.89; 95%CI, 0.61-1.28 for CT or CC vs TT; adjusted OR, 1.14; 95%CI, 0.86-1.79 for TA or AA vs TT). Stratification analysis of two SNPs for risk by subsites only found that variant IL-8-251 TA or AA genotype was associated with increased noncardia GC risk (adjusted OR, 2.58; 95%CI, 1.19-5.57). We did not observe interactions between the IL-8-251 T>A genotype and smoking (adjusted OR, 0.38; 95%CI, 0.08-1.79) or drinking (adjusted OR, 0.36; 95%CI, 0.08-1.65) for risk of noncardia GC. Conclusions: Our data indicate no association between the two SNPs of IL-4-590 and IL-8-251 with overall GC risk, while the IL-8-251 TA or AA genotype conferred risk of cardia GC. Our findings contribute to the evidence body for risk of SNPs associated with the development of gastric cancer in this region.

• BMB Reports
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• 제46권10호
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• pp.479-483
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• 2013

The balance between osteoblast-dependent bone formation and osteoclast-dependent bone resorption maintains bone homeostasis. In inflammatory conditions, this balance shifts toward bone resorption, causing osteolytic bone lesions observed in rheumatoid arthritis and periodontitis. A recently discovered family of cytokine IL-17 is widely reported to mediate diverse inflammatory processes. During the last decade, novel roles for IL-17 in skeletal homeostasis have been discovered indicating the potential importance of this cytokine in bone metabolism. This review will summarize and discuss the involvement of IL-17 during bone homeostasis in both physiologic and pathologic conditions. A better understanding of the role of IL-17 in skeletal systems warrants an advance in bone biology, as well as development of therapeutic strategies against bone-lytic diseases, such as rheumatoid arthritis and periodontitis.