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Immune Regulatory Function of Cancer-Associated Fibroblasts in Non-small Cell Lung Cancer

  • Hyewon Lee;Mina Hwang;Seonae Jang;Sang-Won Um
    • Tuberculosis and Respiratory Diseases
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    • v.86 no.4
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    • pp.304-318
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    • 2023
  • Background: Cancer-associated fibroblasts (CAFs) are key components of the tumor microenvironment and significantly contribute to immune evasion. We investigated the effects of CAFs on the immune function of CD4+ and CD8+ T cells in non-small cell lung cancer (NSCLC). Methods: We isolated CAFs and normal fibroblasts (NFs) from tumors and normal lung tissues of NSCLC patients, respectively. CAFs were co-cultured with activated T cells to evaluate their immune regulatory function. We investigated the effect of CAF conditioned medium (CAF-CM) on the cytotoxicity of T cells. CAFs were also co-cultured with activated peripheral blood mononuclear cells and further incubated with cyclooxygenase-2 (COX2) inhibitors to investigate the potential role of COX2 in immune evasion. Results: CAFs and NFs were isolated from the lung tissues (n=8) and lymph nodes (n=3) of NSCLC patients. Immune suppressive markers, such as COX2 and programmed death-ligand 1 (PD-L1), were increased in CAFs after co-culture with activated T cells. Interestingly, CAFs promoted the expression of programmed death-1 in CD4+ and CD8+ T cells, and strongly inhibited T cell proliferation in allogenic and autologous pairs of CAFs and T cells. CAF-CM decreased the cytotoxicity of T cells. COX2 inhibitors partially restored the proliferation of CD4+ and CD8+ T cells, and downregulated the expression of COX2, prostaglandin E synthase, prostaglandin E2, and PD-L1 in CAFs. Conclusion: CAFs promote immune evasion by suppressing the function of CD4+ and CD8+ T cells via their effects on COX2 and PD-L1 in NSCLC. The immunosuppressive function of CAFs could be alleviated by COX2 inhibitors.

Changes of Inorganic Nitrogen and CO2 Evolution Rate on the Decomposition Process of Korean White Pine Needles (잣나무엽(葉)의 초기(初期) 분해과정(分解過程)에 있어서 무기태(無機態) 질소(窒素) 및 CO2 방출속도(放出速度)의 변화(變化))

  • Yi, Myong Jong;Han, Sang Sup;Kim, Jeong Je
    • Journal of Korean Society of Forest Science
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    • v.69 no.1
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    • pp.13-18
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    • 1985
  • Forest soils mixed with organic matters (green needle, flesh needle litter and needle litter in F layer of Pinus koraiensis, and green leaf of Quercus dentata and Q. variabilis) were incubated under a constant $30^{\circ}C({\pm}1)$ for 53 days to measure the changes of inorganic nitrogen and $CO_2$ evolution rate. The results obtained were summarized as follows; 1) In the early incubation period the amounts of total inorganic nitrogen in soils by mixture of organic matters decreased rapidly because of immobilization by microbial uptake, and thereafter their amounts increased with further incubation. 2) The rate of immobilization of organic nitrogen in mixed organic matters was the highest in green needle among green needle, flesh needle litter and needle litter in F layer of P. koraiensis, but lower than that of green leaf of Q. variabilis and Q. dentata. 3) The rates of $CO_2$ evolution from soils mixed with organic matters increased sharply in the early time, and then decreased slowly with increasing time. The order of the $CO_2$ evolution rate was green leaf of Q. variabilis > green leaf of Q. dentata > green needle of P. koraiensis > flesh needle litter of P. koraiensis > needle litter of P. koraiensis in F layer from the largest to the least. 4) Nitrate nitrogen concentrations showed a tendency to increase throughout incubation time, so that their concentrations after 53 days were higher than that of ammonium nitrogen.

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Improving Effects of Platycodon Extracts Jelly on β-amyloid-induced Cytotoxicity and Scopolamine-Induced Cognitive Impairment Animal Models (도라지 추출물 연양갱이 β-amyloid에 의한 세포독성 및 Scopolamine에 의해 유도된 인지능 저하 동물 모델의 개선효과)

  • Oh, Hong-Geun;Kim, Jung-Hoon;Shin, Eun-Hye;Kang, Young-Rye;Lee, Bong-Gun;Park, Sang-Hoon;Moon, Dae-In;Kwon, Lee-Seong;Kim, Yong-Phill;Choi, Min-Hyu;Kim, Ok-Jin;Park, Kwang-Hyun;Lee, Hak-Yong
    • Korean Journal of Plant Resources
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    • v.26 no.4
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    • pp.417-425
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    • 2013
  • The aim of this study was to examine improving effect of Platycodon extracts (PE) and/or Platycodon extracts jelly (PEJ) on cognitive impairment in vitro and in vivo. PC12 (Pheochromocytoma) cells were pretreated with PE for 1hr and than incubated with $50{\mu}M$ amyloid ${\beta}(A{\beta})_{25-35}$ for additional 48hr. Cell viability was assessed by WST-1. Animals for Morris water test and passive avoidance test were divided into normal, control and two Platycodon extracts treated groups that were named Normal (n=7), Control (0 mg/kg, n=7), PE (300 mg/kg, n=7), PEJ (10 g/kg, n=7). Cognitive impairment was induced by scopolamine (1 mg/kg/body weight, i.p.) in the three experimental groups but not the normal group. Pretretment of PE (0.01-1 mg/mL) were not induced cytotoxicity but observed in high dose-treated group (5 and 10 mg/mL) in PC12 cells. Protective effects of PE against $A{\beta}$-induced cytotoxicity were increased in dose dependent manner in PC12 cells. Administration of PE and PEJ were significantly reduced escape latency time on Morris water maze test and passive avoidance test in copolamine-induced cognitive impairment animal model. These results suggest that Platycodon extracts and its related product available to ameliorative purpose for cognitive ability impairments.

Effects of Taurine Supplementation on Heat Shock Protein 70 and In Vitro Protein Syntheses in Liver of Broiler Chicks under Chronic Heat Stress (고온 스트레스 하에 타우린 첨가가 육계 간의 Heat Shock Protein 70 및 In Vitro의 단백질 합성에 미치는 영향)

  • Cho, Eun So Ri;Park, Garng Hee;Shim, Kwan Seob
    • Korean Journal of Poultry Science
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    • v.43 no.4
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    • pp.213-218
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    • 2016
  • This study was conducted to investigate the effect of taurine supplementation on heat shock protein 70 and in vitro protein turnover in broiler chicks under chronic heat stress. Chicks were allocated into 3 groups of 10 birds per group; the control group was maintained at a temperature of $24^{\circ}C$ without taurine (CO group), the heat-stressed group maintained at a temperature of $34^{\circ}C$ without taurine (HO group), and heat-stressed group maintained at a temperature of $34^{\circ}C$ with taurine (HT group). The final body and liver weights of broilers in the HO and HT groups were significantly lower than those of broilers in the CO group (P<0.05). However, these parameters of the broilers in the HT group were significantly higher than those of broilers in the HO group (P<0.05). The heat shock protein 70 (hsp70) concentration in the liver of broilers in the HO group was significantly higher than that of broilers in the CO and HT groups, but the hsp70 concentration in the liver of broilers in the HT group was not different from that of broilers in the CO group. Liver homogenates of 21 day-old broilers were incubated at temperatures of $37^{\circ}C$ and $45^{\circ}C$ to prove the effect of high temperature and taurine on total protein syntheses. Neither high temperature nor taurine supplementation affected protein syntheses in liver homogenates of the broilers. However, the more the temperature increased, the more the degradation rates of cytoplasmic protein in liver homogenates increased; however, taurine supplementation had no effects on the protein syntheses in the liver of the broiler. It is possible that taurine indirectly affected protein turnover via various physiological mechanisms.

Evaluation of Methods for Determining Lime Requirement of Soils (토양(土壤) 석회요구량(石灰要求量) 결정방법 (決定方法)의 실험적(實驗的) 고찰(考察))

  • Joo, Jin-Ho;Ha, Sang-Keon;Um, Myung-Ho;Lim, Hyung-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.21 no.3
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    • pp.280-288
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    • 1988
  • Lime requirement determination methods were experimentally compared and evaluated for the soils with different physico-chemical properties. The selected soils were mainly distributed in paddy field of Kangweon-Province; Anmi series from limestone region, Dongsong series from basalt region, Gyuam and Gangseo series from alluvial soil. The results were as follows: 1. Differences of soil lime requirement among seven methods ($CaCO_3$ incubation method, $BaCl_2$-TEA method, SMP-single buffer method, Double buffer method, Adams and Evans method, SMP-double buffer method, and O. R. D. method) were remarkably appeared. 2. Measuring lime values by $CaCO_3$ incubation method which is fixed on the basis of lime requirement, SMP-double buffer method was most acceptable for selected soils except Gyuam series, while $BaCl_2$-TEA method showed the highest value, and O. R. D. method was the lowest. 3. Exchangeable Al content of soils was neutralixed near to 70%, but 30% of extractable reached to neutralize when incubated with 100% lime equivalence. 4. Lime requirements based on exchangeable and extractable Al contents of soils were lower than that of $CaCO_3$ incubation method.

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AMPKα, C/EBPβ, CPT1β, GPR43, PPARγ, and SCD Gene Expression in Single- and Co-cultured Bovine Satellite Cells and Intramuscular Preadipocytes Treated with Palmitic, Stearic, Oleic, and Linoleic Acid

  • Choi, S.H.;Park, S.K.;Johnson, B.J.;Chung, K.Y.;Choi, C.W.;Kim, K. H.;Kim, W.Y.;Smith, S.B.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.3
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    • pp.411-419
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    • 2015
  • We previously demonstrated that bovine subcutaneous preadipocytes promote adipogenic gene expression in muscle satellite cells in a co-culture system. Herein we hypothesize that saturated fatty acids would promote adipogenic/lipogenic gene expression, whereas mono- and polyunsaturated fatty acids would have the opposite effect. Bovine semimembranosus satellite cells (BSC) and intramuscular preadipocytes (IPA) were isolated from crossbred steers and cultured with 10% fetal bovine serum (FBS)/Dulbecco's Modified Eagle Medium (DMEM) and 1% antibiotics during the 3-d proliferation period. After proliferation, cells were treated for 3 d with 3% horse serum/DMEM (BSC) or 5% FBS/DMEM (IPA) with antibiotics. Media also contained $10{\mu}g/mL$ insulin and $10{\mu}g/mL$ pioglitazone. Subsequently, differentiating BSC and IPA were cultured in their respective media with $40{\mu}M$ palmitic, stearic, oleic, or linoleic acid for 4 d. Finally, BSC and IPA were single- or co-cultured for an additional 2 h. All fatty acid treatments increased (p = 0.001) carnitine palmitoyltransferase-1 beta ($CPT1{\beta}$) gene expression, but the increase in $CPT1{\beta}$ gene expression was especially pronounced in IPA incubated with palmitic and stearic acid (6- to 17-fold increases). Oleic and linoleic acid decreased (p = 0.001) stearoyl-CoA desaturase (SCD) gene expression over 80% in both BSC and IPA. Conversely, palmitic and stearic acid increased SCD gene expression three fold in co-cultured in IPA, and stearic acid increased $AMPK{\alpha}$ gene expression in single- and co-cultured BSC and IPA. Consistent with our hypothesis, saturated fatty acids, especially stearic acid, promoted adipogenic and lipogenic gene expression, whereas unsaturated fatty acids decreased expression of those genes associated with fatty acid metabolism.

Gene Transfer into Pig and Goat Fetal Fibroblasts by Co-transfection of tPA Transgene and $Neo^r$ Gene

  • Kim, Bae-Chul;Han, Rong-Xun;Kim, Myung-Yoon;Shin, Young-Min;Park, Chang-Sik;Jin, Dong-Il
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.107-111
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    • 2009
  • The transfection efficiency of a transgene into pig and goat fetal fibroblast cells (PFF and GFF, respectively) was tested using co-transfection of a human tissue-type plasminogen activator (tPA) transgene and neomycin-resistant ($Neo^r$) gene, followed by G418 selection. To initially test G418 resistance, GFF and PFF were incubated in culture medium containing different concentration of G418 for 2 weeks, and cell survival was monitored over time. Based on the obtained results, the concentrations chosen for G418 selection were 800 ug/ml and 200 ug/ml for GFF and PFF, respectively. For co-transfection experiments, the pBC1/tPA and $Neo^r$ vectors were co-transfected into GFF and PFF ($1{\times}10^6$ cells in each case) using the FuGENE6 transfection reagent, and resistant colonies were obtained following 14 days of G418 selection. We obtained 96 and 93 drug-resistant colonies of GFF and PFF, respectively, only 54 and 39 of which, respectively, continued proliferating after drug selection. PCR-based screening revealed that 23 out of 54 analyzed GFF colonies and 5 out of 39 analyzed PFF colonies contained insertion of the tPA gene. Thus, the experimentally determined transfection efficiencies for tPA gene co-transfection with the $Neo^r$ gene were 42.6% for GFF and 12.8% for PFF. These findings suggest that co-transfection of a transgene with the $Neo^r$ gene can aid in the successful integration of the transgene into fetal fibroblast cells.

Biomaterials Inhibiting Biofilm Formation of Staphylococcus aureus (생물소재를 이용한 황색포도상구균의 바이오필름 억제 연구)

  • Shin, Kye-Ho;Yun, Yu-Na;Jeon, Gi-Boong;Lee, Tae-Ryong;Yi, Sung-Won;Cho, Jun-Cheol;Park, Ji-Yong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.4
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    • pp.347-350
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    • 2011
  • Biofilms are surface-attached microbial communities with phenotypic and biochemical properties distinct from free-living planktonic cells. Biofilm bacteria show much greater resistance than planktonic counterparts and much higher concentration of biocide is needed to treat biofilms compared to the dosage used for planktonic bacteria. As a result, alternative strategies or more effective agents exhibiting activity against biofilm-producing micro-organisms are of great interest. Therefore, we turned our attention to control of biofilm of S. aureus. The aims of this research are to investigate substances which inhibit the formation of biofilm by S. aureus and to suggest effective materials for controlling skin problems. We coated slide glasses with human placental collagen and the coverslip was incubated with test materials and bacteria. The coverslip was stained with crystal violet and we measured optical density of each sample. The biofilm inhibitory activity was calculated by crystal violet staining degrees. In this study, S. aureus ATCC 6538 was used as test organism. Our results show that both water soluble and insoluble Hinoki cypress polysaccharide strongly inhibited biofilm formation. Whereas, green tea and sunset hibiscus root extract promoted biofilm. Xylitol showed a concentration dependent effect; high concentration (3 % and 5 %) of xylitol reduced biofilm while promoted biofilm formation at a concentration of 1 %. These results support that Hinoki cypress polysaccharide and xylitol have ability to suppress biofilm formation.

High Glucose Induces Apoptosis through Caspase-3 Dependent Pathway in Human Retinal Endothelial Cell Line (인간망막 내피세포주에서 고농도 포도당이 caspase-3 경로를 통해 세포자연사 유도)

  • Seo, Eun-Sun;Chae, Soo-Chul;Kho, Eun-Gyeong;Lee, Jong-Bin
    • Korean Journal of Environmental Biology
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    • v.27 no.1
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    • pp.66-72
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    • 2009
  • Diabetic Retinopathy (DR) is a leading cause of blindness among adults in the western countries. Hyperglycemia is a condition, that induces apoptotic cell death in a variety of cell types in diabetes, but the mechanism remains unclear. The aim of the study is to understand the effects of high Glucose on Human Retinal Endothelial Cells. Retinal endothelial cells were cultured in Iscove's Modified Dulbecco's Medium (IMDM) containing 5, 25 and 50 mM Glucose, incubated for 24, 36 and 48 hours in humidified 5 % CO$_2$ incubator at 37$^{\circ}C$. Human Retinal Endothelial Cell Line (HREC) were characterized for morphology with different treatment by phase contrast microscopic analysis. Number of dead and viable cells was counted by trypan blue exclusion and supported by MTT assay. The intracellular Hydrogen peroxide (H$_2$O$_2$), a Reactive Oxygen Species (ROS) generation in high glucose conditions was assessed by FOX II assay and apoptosis by caspase-3 assay. The high glucose treated cells undergoing DNA fragmentation was witnessed by Agarose gel electrophoresis. We found that the cells incubated with 25 and 50 mM glucose containing medium for 48 hours altered the morphology of the cell, induced apoptosis and DNA fragmentation. The dead cell number were high in 25 and 50 mM when compared to the cells incubated with 5 mM glucose for 24, 36, and 48 hours. Also, the H$_2$O$_2$ levels and the activity of caspase-3 were increased in high glucose treated cells. Conclusions/interpretation: Our results demonstrated that elevated glucose induces apoptosis in cultured HREC. The hyperglycemia-induced increase in apoptosis may be dependent on caspase activation. The association between ROS generation and caspase-3 activation on high glucose treated cells is yet to be investigated.

Anti-atherosclerotic Effects of Herbal Formulas for Sasang Constitutional Medicine (사상 체질 처방의 항동맥경화 효능 비교 연구 -열다한소탕, 양격산화탕, 청심연자탕, 태음조위탕-)

  • Kim, Ohn-Soon;Kim, Ye-Ji;Shin, Hyeun-Kyoo
    • Journal of Sasang Constitutional Medicine
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    • v.24 no.4
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    • pp.51-61
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    • 2012
  • Objectives : The purpose of this study was to investigate the anti-atherosclerotic effects of 4 herbal formulas for Sasang constitutional medicine (Yeoldahanso-tang: YDHST, Yanggyeoksanhwa-tang: YGSHT, Cheongsimyeonja-tang: CSYJT and Taeeumjowi-tang: TEJWT). Methods : The antioxidant activities of herbal formulas were studied by measuring free radical scavenging activities on ABTS and DPPH. The inhibitory effects on LDL oxidation was evaluated by the formation of TBARS, REM and fragmentation of apolipoprotein B-100 (ApoB). Effects of herbal formulas on macrophage lipid accumulation were determined in native LDL and LPS co-incubated macrophages using Oil Red O staining. Results : The scavenging activities on ABTS and DPPH of herbal formulas were increased in dose-dependent manner (YDHST>YGSHT>CSYJT>TEJWT). Herbal formulas reduced the oxidation properties of LDL induced by $CuSO_4$. YDHST, YGSHT and CSYJT showed strong suppressive effect on LDL oxidation than TEJWT. In addition, YDHST, YGSHT and CSYJT significantly inhibited foam cell formation in LDL/LPS stimulated RAW 264.7 cells. Conclusions : These results demonstrate that YDHST, YGSHT and CSYJT have potentials on anti-atherosclerosis by antioxidative effect and suppressive effect on LDL oxidation.