• Title/Summary/Keyword: Cell treatment

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Synovial Cell Migration is Associated with B Cell Activating Factor Expression Increased by TNFα or Decreased by KR33426

  • Lee, Jiyoung;Yoon, Sung Sik;Thuy, Pham Xuan;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.28 no.5
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    • pp.405-413
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    • 2020
  • Fibroblast-like synoviocytes (FLS) play a crucial role in initiating rheumatoid arthritis. B-cell activating factor (BAFF) plays a role in FLS survival as well as in B cell maturation and maintenance. Here, we investigated whether tumor necrosis factor (TNF)-α-induced BAFF expression controls FLS migration and whether BAFF expression in FLS could be regulated by KR33426 which is the inhibitor of BAFF binding to BAFF receptors (BAFF-R) by using MH7A synovial cells transfected with the SV40 T antigen. More TNF-α-treated cells migrated compared to the control. TNF-α increased BAFF expression in FLS, significantly. FLS migration was inhibited by the transfection with BAFF-siRNA. KR33426 also inhibited BAFF expression increased by TNF-α treatment in FLS as judged by western blotting, PCR, and transcriptional activity assay. Kinases including JNK, p38 and Erk were activated by TNF-α treatment. While JNK and p38 were inhibited by KR33426 treatment, no changes in Erk were observed. Transcription factors including p65, c-Fos, CREB and SP1 were enhanced by TNF-α treatment. Among them, c-Fos was inhibited by KR33426 treatment. Small interference(si)-RNA of c-fos decreased BAFF transcriptional activity. FLS migration induced by TNF-α was inhibited by the transfection with BAFF-siRNA. KR33426 increased Twist, Snail, Cadherin-11 and N-Cadherin. In contrast, KR33426 decreased E-cadherin and TNF-α-enhanced CCL2. Taken together, our results demonstrate that synovial cell migration via CCL2 expression could be regulated by BAFF expression which is decreased by KR33426 and c-Fos-siRNA. It suggests for the first time that the role of BAFF-siRNA on FLS migration might be matched in the effect of KR33426 on BAFF expression.

Effects of Endoplasmic Reticulum Stress Inhibitor Treatment during the Micromanipulation of Somatic Cell Nuclear Transfer in Porcine Oocytes

  • Park, Yeo-Reum;Park, Hye-Bin;Kim, Mi-Jeong;Jung, Bae-Dong;Lee, Seunghyung;Park, Choon-Keun;Cheong, Hee-Tae
    • Development and Reproduction
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    • v.23 no.1
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    • pp.43-54
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    • 2019
  • We examined the effects of endoplasmic reticulum (ER) stress inhibitor treatment during the micromanipulation of porcine somatic cell nuclear transfer (SCNT) on the in vitro development of SCNT embryos. ER stress inhibitors such as salubrinal (200 nM) and tauroursodeoxycholic acid (TUDCA; $100{\mu}M$) were added to the micromanipulation medium and holding medium. The expression of X-box binding protein 1 (Xbp1), ER-stress-associated genes, and apoptotic genes in SCNT embryos was confirmed at the one-cell and blastocyst stages. Levels of Xbp1 splicing and expression of ER-stress-associated genes in SCNT embryos at the one-cell stage decreased significantly with TUDCA treatment (p<0.05). The expression of ER-stress-associated genes also decreased slightly with the addition of both salubrinal and TUDCA (Sal+TUD). The expression levels of caspase-3 and Bcl2-associated X protein (Bax) mRNA were also significantly lower in the TUDCA and Sal+TUD treatments (p<0.05). At the blastocyst stage, there were no differences in levels of Xbp1 splicing, and transcription of ER-stress-associated genes and apoptosis genes between control and treatment groups. However, the blastocyst formation rate (20.2%) and mean blastocyst cell number ($63.0{\pm}7.2$) were significantly higher (p<0.05) for embryos in the TUDCA treatment compared with those for control (12.6% and $41.7{\pm}3.1$, respectively). These results indicate that the addition of ER-stress inhibitors, especially TUDCA, during micromanipulation can inhibit cellular damage and enhance in vitro development of SCNT embryos by reducing stress levels in the ER.

The Study on Acute and Subacute Toxicity and Sarcoma-180 Anti-cancer Effects of Triglii Semen Herbal-acupuncture (파두약침(巴豆藥鍼)의 급성(急性) 아급성(亞急性) 독성실험(毒性實驗) 및 Sarcoma-180 항암효과(抗癌效果)에 관(關)한 실험적(實驗的) 연구(硏究))

  • Yoo, Chang-Kil;Kwon, Ki-Rok;Yu, Byeong-Gil
    • Journal of Pharmacopuncture
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    • v.5 no.1 s.8
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    • pp.27-42
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    • 2002
  • Objective: The purpose of this study was to investigate the acute and subacute toxicity and sarcoma- 180 anti-cancer effects of Herbal acupuncture with Triglii Semen in mice and rats. Method: Balble mice were injected intraperitoneally with Triglii semen Herbal acupuncture for $LD_{50}$ and acute toxicity test. Sprague Dawley rats were injected intraperitoneally with Triglii semen Herbal acupuncture for subacute toxicity test. The Triglii semen Herbal acupuncture was injected on Chung-wan(CV12) of mice with S-180 cancer cell line. Results: 1. In acute toxicity test, the $LD_{50}$ value was $7.49{\times}10^3$ml, 0.30ml/kg.2. The body weights of mice treated with Triglii semen Herbal acupuncture increased during the acute toxicity test. 3. In acute toxicity test of serum biochenrical values of mice, total protein was decreased in treatment groups I, 2 and 3, albunrin was decreased in treatment groups 2 and 3 compared to the control group. GOT was increased in treatment group I and Alk. Phosphatase was increased in treatment groups 1,2 and 3 compared to the normal group(p<0.05). 4.ln subacute toxicity test, severe tissue injury was found in lung and liver. 5. In subacute toxicity test, the body weight was decreased in treatment groups I and 2 compared to the normal group and the weight of liver. lung and kidney were increased in treatment groups 1, 2 and 3 compared to the normal group.(p<0.05) 6. In subacute toxicity test, RBC, HGB and HCT were decreased in treatment groups 1 and 2 compared to the normal group. MCV was increased in treatment group1 compared to the normal group, MCH was increased in treatment groups 1 and 2 compared to the control group in complete blood count test.(p<0.05) 7. In subacute toxicity test, total protein was decreased in treatment groups 1 and 2 compared to the nonnal group, BUN was increased in treatment groups 1 and 2 compared to the nonnal group, creatinine and uric acid were decreased in treatment groups 1 and 2 compared to the normal group, glucose was increased in treatment group 2 compared to the nonnal group, triglycelide was decreased in treatment groups I and 2 compared to the normal group, total cholesterol was increased in treatment groups 1 and 2 compared to the control group. GOT was decreased in treatment group 2 compared to the normal and control group, AIk. Phosphatase was increased in treatment group 1 compared to the normal and control group.(p<0.05) 8. Median survival time was 17days in treatment group 2 for S- 180 cancer cell treated with Triglii semen Herbal acupuncture. 9. Natural killer cell activity was insignificant for S-180 cell treated with Triglii semen Herbal acupuncture.(p<0.05) 10. lnterieukin-2 productivity was decreased for S-180 cell treated with Triglii semen Herbal acupuncture compared to the normal and control group.(p<0.05) Conclusion: According to the results, we can conclude Herbal-acupuncture with Triglii semen caused toxicity, and caused no effects in S-180 cancer cell.

Recombinant Human Thioredoxin-1 Protects Macrophages from Oxidized Low-Density Lipoprotein-Induced Foam Cell Formation and Cell Apoptosis

  • Zhang, Hui;Liu, Qi;Lin, Jia-Le;Wang, Yu;Zhang, Ruo-Xi;Hou, Jing-Bo;Yu, Bo
    • Biomolecules & Therapeutics
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    • v.26 no.2
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    • pp.121-129
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    • 2018
  • Oxidized low-density lipoprotein (ox-LDL)-induced macrophage foam cell formation and apoptosis play critical roles in the pathogenesis of atherosclerosis. Thioredoxin-1 (Trx) is an antioxidant that potently protects various cells from oxidative stress-induced cell death. However, the protective effect of Trx on ox-LDL-induced macrophage foam cell formation and apoptosis has not been studied. This study aims to investigate the effect of recombinant human Trx (rhTrx) on ox-LDL-stimulated RAW264.7 macrophages and elucidate the possible mechanisms. RhTrx significantly inhibited ox-LDL-induced cholesterol accumulation and apoptosis in RAW264.7 macrophages. RhTrx also suppressed the ox-LDL-induced overproduction of lectin-like oxidized LDL receptor (LOX-1), Bax and activated caspase-3, but it increased the expression of Bcl-2. In addition, rhTrx markedly inhibited the ox-LDL-induced production of intracellular reactive oxygen species (ROS) and phosphorylation of p38 mitogen-activated protein kinases (MAPK). Furthermore, anisomycin (a p38 MAPK activator) abolished the protective effect of rhTrx on ox-LDL-stimulated RAW264.7 cells, and SB203580 (a p38 MAPK inhibitor) exerted a similar effect as rhTrx. Collectively, these findings indicate that rhTrx suppresses ox-LDL-stimulated foam cell formation and macrophage apoptosis by inhibiting ROS generation, p38 MAPK activation and LOX-1 expression. Therefore, we propose that rhTrx has therapeutic potential in the prevention and treatment of atherosclerosis.

The behavior characteristics of immobilized sludge in waste water treatment using sequencing batch reactor(SBR). (연속 회분식 반응기를 이용한 폐수처리에서 고정화 슬러지의 거동 특성)

  • 최석순
    • Journal of environmental and Sanitary engineering
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    • v.11 no.2
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    • pp.1-7
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    • 1996
  • The behavior of total organic carbon (TOC) and phosphate were observed for 15 days with immobilized activated sludge using polyacrylamide (PAA) by sequencing batch reactor (SBR). In the preparation of immobilized sludge by PAA, it was found that suitable acrylamide concentration for actual wastewater treatment was to be 15% through the batch test. When SBR system was operated in the repeated aerobic and anaerobic conditions, TOC removal efficiency was 92%. The uptake rate of phosphate was increased from 1.78 mg-P/g cell/hr on the 5th day of acclimation to 2.5 mg-P/g cell/hr on the 15th day of acclimation. And the total phosphorus content in PAA bead was increased from 40 mg-P/g cell on the 1st day of operation to 55 mg-P/g cell on the 15th day of operation. From this study, lowering the volume of aeration tank was possible when PAA bead was used in wastewater treatment and long operation was also possible without the settler.

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The Role of Protein Kinase C for Prolactin Secretion in Chicken Primary Pituitary Cell Culture (산란계의 뇌하수체 세포배양에서 Prolactin의 생성에 관계하는 Protein Kinase C의 역할)

  • 선상수
    • Korean Journal of Poultry Science
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    • v.23 no.3
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    • pp.113-119
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    • 1996
  • A series of experiments were conducted to investigate the role of protein kinase C (PKC) as a second messenger in vasoactive intestinal peptide (VIP) mediated prolactin secretion. Primary pituitary cells (106 cells/treatment) were separated from laying hens and incubated in M-199 with 5% chicken serum and 5% fetal calf serum. The VIP(0.1 $\pi$M) treatment enhanced prolactin Secretion into media upto 9-fold during 48-h incubation. The phorbol 12-myristate 13-acetate (PMA), a PKG agonist, increased prolactin secretion upto 2-fold at 0.1 nM PMA (P<0.01), and the prolactin secretion was not significantly higher than this concentration. Staurosporine (ST; 1.0$\pi$M) a PKC antagonist, decreased by 70% of 0.1 $\pi$M VIP-stimulated prolactin secretion and by 48% of 10 ${\mu}$M PMA-stimulated prolactin secretion (P<0.01). However, pituitary cell prolactin content did not differ in any treatment (P>0.05). In conclusion, these results indicate that the PKC second messenger system is involved in VIP-stimulated prolactin release in chicken primary pituitary cell culture.

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Chloroquine and Valproic Acid Combined Treatment in Vitro has Enhanced Cytotoxicity in an Osteosarcoma Cell Line

  • Wang, Chuan-Kun;Yu, Xi-Dong;Li, Qiang;Xie, Gang;Teng, Yue
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4651-4654
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    • 2013
  • Choroquine (CQ) and valproic acid (VPA) have been extensively studied for biological effects. Here, we focused on efficacy of combined CQ and VPA on osteosarcoma cell lines. Viability of osteosarcoma cell lines (U20S and HOS) was analyzed by MTT assay. Apoptotic assays and colony formation assays were also applied. ROS generation and Western Blotting were performed to determine the mechanism of CQ and VPA combination in the process of apoptosis. The viability of different osteosarcoma cell lines significantly decreased after CQ and VPA combination treatment compared with either drug used alone, and apoptosis was increased significantly. ROS generation was triggered leading to expression of apoptosis related genes being increased and of antiapoptotic related genes being decreased. From our data shown here, CQ and VPA combination treatment in vitro enhanced cytotoxicy to osteosarcoma cells.

Electric power generation from treatment of food waste leachate using microbial fuel cell

  • Wang, Ze Jie;Lim, Bong Su
    • Environmental Engineering Research
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    • v.22 no.2
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    • pp.157-161
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    • 2017
  • Simultaneous treatment of food waste leachate and power generation was investigated in an air-cathode microbial fuel cell. A TCOD removal efficiency of $95.4{\pm}0.3%$ was achieved for an initial COD concentration of 2,860 mg/L. Maximum power density ranged was maximized at $1.86W/m^3$, when COD concentration varied between 60 mg/L and 2,860 mg/L. Meanwhile, columbic efficiency was determined between 1.76% and 11.07% for different COD concentrations. Cyclic voltammetric data revealed that the oxidation peak voltage occurred at -0.20 V, shifted to about -0.25 V. Moreover, a reduction peak voltage at -0.45 V appeared when organic matters were exhausted, indicating that reducible matters were produced during the decomposition of organic matters. The results showed that it was feasible to use food waste leachate as a fuel for power generation in a microbial fuel cell, and the treatment efficiency of the wastewater was satisfied.

Anticancer Effects of Gut Microbiota-Derived Short-Chain Fatty Acids in Cancers

  • Mi-Young Son;Hyun-Soo Cho
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.849-856
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    • 2023
  • Short-chain fatty acids (SCFAs), such as butyrate, propionate, and acetate produced by the gut microbiota have been implicated in physiological responses (defense mechanisms, immune responses, and cell metabolism) in the human body. In several types of cancers, SCFAs, especially butyrate, suppress tumor growth and cancer cell metastasis via the regulation of the cell cycle, autophagy, cancer-related signaling pathways, and cancer cell metabolism. In addition, combination treatment with SCFAs and anticancer drugs exhibits synergistic effects, increasing anticancer treatment efficiency and attenuating anticancer drug resistance. Therefore, in this review, we point out the importance of SCFAs and the mechanisms underlying their effects in cancer treatment and suggest using SCFA-producing microbes and SCFAs to increase therapeutic efficacy in several types of cancers.

Vemurafenib Enhances NK cell Expansion and Tumor-killing Activity for Cancer Immunotherapy

  • Min Hwa Shin
    • Biomedical Science Letters
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    • v.29 no.4
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    • pp.371-375
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    • 2023
  • Natural killer (NK) cells are innate immune cells and play important roles as the first immune cells to recognize and kill cancer. In patients with advanced and terminal cancer, NK cells are often inactivated, suggesting that NK cells may play important roles in cancer treatment. In particular, the proportion of NK cells among immune cells infiltrating tumor tissues is often low, which suggests that NK cells do not survive in tumor microenvironment (TME). In order to overcome these hurdles of NK cells in cancer treatment, it is critical to develop strategies that enhance the proliferation and cytolytic activity of NK cells. We applied Vemurafenib to NK cells and measured the degree of NK cell proliferation and functional activation. We obtained unexpected results of increased NK cell numbers and anti-tumor activity after Vemurafenib treatment. Although further investigation is required to uncover the detailed mechanisms, our results suggest that Vemurafenib is a promising candidate to increase the efficacy of cancer immunotherapy using NK cells.