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Anti-apoptotic effect by Bcl-2 in UVB-irradiated keratinocytes.

  • Takahashi, Hidetoshi;Honma, Masaru;Ishida-Yamamoto, Akemi;Namikawa, Kazuhiko;Miwa, Akiko;Okado, Haruo;Kiyama, Hiroshi;Iizuka, Hajime
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.225-228
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    • 2002
  • Bcl-2 is a member of large bcl-2 family and protects cells from apoptosis. Using bcl-2-expressing adenovirus vector (Ad-bc1-2), we investigated the effect of bc1-2 on UVB-induced apoptosis. Adenovirus vector efficiently introduced bc1-2 gene in cultured normal mouse keratinocytes (NMK cells); almost all NMK cells (lx10$^{6}$ ) were transfected at Ixl0$^{8}$ PFU/ml. Bcl-2-transfected NMK cells were significantly resistant to UVB-induced apoptosis with the suppressive effect dependent on bcl-2-expression level. Following UVB irradiation caspase 8, 3, 9 activities were stimulated in NMK cells, while in bc1-2-transfected cells, only caspase 8, but not caspase 3 or 9 activities were stimulated. In order to investigate the effect of bc1-2 in vivo, topical application of Ad-bc1-2 on tape-stripped mouse skin was performed. Following the application, Bc1-2 was efficiently overexpressed in almost all viable keratinocytes. The expression was transient with the maximal expression of Bc1-2 at 1st day following the application of lxl0$^{9}$ PFU in 200ml. The introduced Bc1-2 remained at least for 6 days. UVB irradiation (1250 J/m$^2$) induced apoptosis within 12 h and the maximal effect was observed at 24 h in control mouse skin. Bc1-2-transfected mice skin were resistant to UVB-induced apoptosis. Topical application of empty adenovirus vector alone had no effect on Bc1-2 expression or UVB-induced apoptosis. These results indicate that adenovirus vector is an efficient gene delivery system into keratinocytes and that Bcl-2 is a potent inhibitor of UVB-induced apoptosis both in vitro and in vivo.

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Low-Level Expression of CD138 Marks Naturally Arising Anergic B Cells

  • Sujin Lee;Jeong In Yang;Joo Hee Lee;Hyun Woo Lee;Tae Jin Kim
    • IMMUNE NETWORK
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    • v.22 no.6
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    • pp.50.1-50.19
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    • 2022
  • Autoreactive B cells are not entirely deleted, but some remain as immunocompetent or anergic B cells. Although the persistence of autoreactive B cells as anergic cells has been shown in transgenic mouse models with the expression of B cell receptor (BCR) reactive to engineered self-antigen, the characterization of naturally occurring anergic B cells is important to identify them and understand their contribution to immune regulation or autoimmune diseases. We report here that a low-level expression of CD138 in the splenic B cells marks naturally arising anergic B cells, not plasma cells. The CD138int B cells consisted of IgMlowIgDhigh follicular (FO) B cells and transitional 3 B cells in homeostatic conditions. The CD138int FO B cells showed an anergic gene expression profile shared with that of monoclonal anergic B cells expressing engineered BCRs and the gene expression profile was different from those of plasma cells, age-associated B cells, or germinal center B cells. The anergic state of the CD138int FO B cells was confirmed by attenuated Ca2+ response and failure to upregulate CD69 upon BCR engagement with anti-IgM, anti-IgD, anti-Igκ, or anti-IgG. The BCR repertoire of the CD138int FO B cells was distinct from that of the CD138- FO B cells and included some class-switched B cells with low-level somatic mutations. These findings demonstrate the presence of polyclonal anergic B cells in the normal mice that are characterized by low-level expression of CD138, IgM downregulation, reduced Ca2+ and CD69 responses upon BCR engagement, and distinct BCR repertoire.

Anti-cancer Effects and Changes in Colonic Microflora of Polysaccharide Derived from Edible Mushroom Mycelium on AOM/DDS-induced Colon Cancer Model (AOM/DDS로 대장암 유도 후 식용버섯균사체 유래 다당류의 대장암 억제효과 및 대장 미생물균총의 변화)

  • Seaung Sik Kong;Soon Ah Kang
    • The Korean Journal of Food And Nutrition
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    • v.37 no.3
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    • pp.139-151
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    • 2024
  • The objective of this study was to investigate the anticancer effects of EMPS (edible mushroom mycelium polysaccharide: Tremella fuciformis) in animal models with colorectal cancer induced by AOM/DSS. The experimental groups consisted of Nor (normal), NC (AOM/DSS), EMPS (EMPS 50, EMPS 100), and PC (Fluorouracil). The NC group had the highest number of colon tumors, whereas it was observed that tumor occurrence was significantly reduced in the EMPS consumption group. The expression of Bcl-2, an apoptosis inhibitor, was significantly lower in the EMPS 50 & 100 and PC groups. On the other hand, the mRNA gene expression of Bax, a factor that induces apoptosis, was significantly higher in the EMPS 50 & 100 and PC groups compared to the NC group. The mRNA expression levels of TNF-α and COX-2 significantly increased in the NC group, but showed a significant decrease in the EMPS and PC groups, indicating inhibition of the cancer-promoting response of cells. At the phylum level of the mice's intestinal microbial composition, the proportion of Bacteroidetes tended to decrease, while the proportion of Firmicutes tended to increase with EMPS administration. This suggests that changes in the gut microbiota caused by inflammation can be influenced by dietary intake.

ppGalNAc T1 as a Potential Novel Marker for Human Bladder Cancer

  • Ding, Ming-Xia;Wang, Hai-Feng;Wang, Jian-Song;Zhan, Hui;Zuo, Yi-Gang;Yang, De-Lin;Liu, Jing-Yu;Wang, Wei;Ke, Chang-Xing;Yan, Ru-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5653-5657
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    • 2012
  • Objectives: To investigate the effect of glycopeptide-preferring polypeptide GalNAc transferase 1 (ppGalNAc T1 ) targeted RNA interference (RNAi) on the growth and migration of human bladder carcinoma EJ cells in vitro and in vivo. Methods: DNA microarray assays were performed to determine ppGalNAc Ts(ppGalNAc T1-9) expression in human bladder cancer and normal bladder tissues. We transfected the EJ bladder cancer cell line with well-designed ppGalNAc T1 siRNA. Boyden chamber and Wound healing assays were used to investigate changes of shppGalNAc T1-EJ cell migration. Proliferation of shppGalNAc T1-EJ cells in vitro was assessed using [3H]-thymidine incorporation assay and soft agar colony formation assays. Subcutaneous bladder tumors in BALB/c nude mice were induced by inoculation of shppGalNAc T1-EJ cells and after inoculation diameters of tumors were measured every 5 days to determine gross tumor volumes. Results: ppGalNAc T1 mRNA in bladder cancer tissues was 11.2-fold higher than in normal bladder tissues. When ppGalNAc T1 expression in EJ cells was knocked down through transfection by pSUPER-shppGalNAc T1 vector, markedly reduced incorporation of [3H]-thymidine into DNA of EJ cells was observed at all time points compared with the empty vector transfected control cells. However, ppGalNAc T1 knockdown did not significantly inhibited cell migration (only 12.3%). Silenced ppGalNAc T1 expression significantly inhibited subcutaneous tumor growth compared with the control groups injected with empty vector transfected control cells. At the end of observation course (40 days), the inhibitory rate of cancerous growth for ppGalNAc T1 knockdown was 52.5%. Conclusion: ppGalNAc T1 might be a potential novel marker for human bladder cancer. Although ppGalNAc T1 knockdown caused no remarkable change in cell migration, silenced expression significantly inhibited proliferation and tumor growth of the bladder cancer EJ cell line.

Hepatoprotective Effects of Lactic Acid-Fermented Garlic Extracts on Fatty Liver-Induced Mouse by Alcohol (알코올성 지방간을 유발시킨 마우스에서 유산균 발효 마늘추출물의 간 보호 효과)

  • Choi, Ji-Hwi;Kim, Min-Seok;Yu, Heui-Jong;Kim, Ki-Ho;Lee, Hee-Seop;Cho, Hong-Yon;Lee, Seung-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.11
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    • pp.1642-1647
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    • 2014
  • In this study, we investigated the hepatoprotective effects of garlic extracts fermented with Lactobacillus plantarum on alcohol-induced fatty liver damage in C57BL/6J mice. The weight of liver tissue of the lactic acid-fermented garlic extract (LAFGE) diet groups decreased in a dose-dependent manner compared to that of the normal diet group. The activities of typical serum enzymes such as ALT, AST, and ALP were low in the LAFGE 200 mg/kg administered group. The LAFGE 200 mg/kg administered group significantly decreased values of TBIL and DBIL, which are an important index of liver damage. LAFGE also ameliorated alcohol-induced hepatic lipid accumulation in histological analysis dose-dependently. In addition, total GSH and reduced GSH levels in LAFGE-treated groups were gradually recovered up to normal levels. The present study revealed that LAFGE improved unique tastes and odors of raw garlic through lactic acid fermentation technology and thus could be a commercially potential material for hepatoprotective functional foods against alcoholic fatty liver.

A Cancer-specific Promoter for Gene Therapy of Lung Cancer, Protein Regulator of Cytokinesis 1 (PRC1) (폐암의 유전자 치료법을 위한 암특이적인 PRC1 프로모터)

  • Cho, Young-Hwa;Yun, Hye-Jin;Kwon, Hee-Chung;Kim, Hee-Jong;Cho, Sung-Ha;Kang, Bong-Su;Kim, Yeun-Ju;Seol, Won-Gi;Park, Kee-Rang
    • Journal of Life Science
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    • v.18 no.10
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    • pp.1395-1399
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    • 2008
  • We have recently reported the PRC1 promoter as a promoter candidate to control expression of transcriptionally targeted genes for breast cancer gene therapy. We tested whether the PRC1 promoter could be also applied for the lung cancer gene therapy. In the transient transfection assay with naked plasmids containing the luciferase fused to the PRC1 promoter, the promoter showed little activity in the normal lung cell line, MRC5. However, in the lung cancer A549 cells, PRC1 showed approximately 30-fold activation which was similar to the survivin promoter, the gene whose promoter has been already reportedas a candidate for the gene therapy of lung cancer. In viral systems, the PRC1 promoter showed approximately 75% and 66% of transcriptional activity compared to the CMV promoter in the adeno-associated virus (AAV) and the adenovirus (AV) systems, respectively. However, the PRC1 promoter in either AAV or AV showed approximately 20% activity compared to the CMV promoter in the normal lung cells. In addition, human lung tumor xenograft mice showed that the PRC1 promoter activity was as strong as the CMV activity in vivo. Taken together, these results suggested that PRC1 might be a potential promoter candidate for transcriptionally targeted lung cancer gene therapy.

A screening of Alzheimer's disease using basis synthesis by singular value decomposition from Raman spectra of platelet (혈소판 라만 스펙트럼에서 특이값 분해에 의한 기저 합성을 통한 알츠하이머병 검출)

  • Park, Aaron;Baek, Sung-June
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.5
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    • pp.2393-2399
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    • 2013
  • In this paper, we proposed a method to screening of Alzheimer's disease (AD) from Raman spectra of platelet with synthesis of basis spectra using singular value decomposition (SVD). Raman spectra of platelet from AD transgenic mice are preprocessed with denoising, removal background and normalization method. The column vectors of each data matrix consist of Raman spectrum of AD and normal (NR). The matrix is factorized using SVD algorithm and then the basis spectra of AD and NR are determined by 12 column vectors of each matrix. The classification process is completed by select the class that minimized the root-mean-square error between the validation spectrum and the linear synthesized spectrum of the basis spectra. According to the experiments involving 278 Raman spectra, the proposed method gave about 97.6% classification rate, which is better performance about 6.1% than multi-layer perceptron (MLP) with extracted features using principle components analysis (PCA). The results show that the basis spectra using SVD is well suited for the diagnosis of AD by Raman spectra from platelet.

Anti-Obesity Effect of Krill Oil by Regulation of Adipokines in High Fat Diet-Induced Mouse Model (고지방식이 동물모델에서 크릴오일의 아디포카인 조절을 통한 항비만 효과)

  • Kim, Ji Hyun;He, Mei Tong;Seo, Hyo Jeong;Lee, Dongjun;Cho, Eun Ju
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.11
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    • pp.201-208
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    • 2020
  • This study examined the anti-obesity effect of krill oil (KO) by regulating adipokines in a high-fat diet (HFD)-induced obese mouse model. The mice were fed a 60 kcal% HFD for 16 weeks, and KO was then administered at an oral dose of 100, 200, and 500 mg/kg/day for four weeks before the end of the experiment. The administration of KO at concentrations of 200 and 500 mg/kg/day decreased body weight gain significantly compared with the HFD-fed control group. In addition, the HFD-fed control group showed the abnormal release of adipokines by an increase in leptin and decrease in adiponectin, compared to the normal diet-fed normal group. On the other hand, KO (500 mg/kg/day)-administered group attenuated the abnormal release of adipokines by the down-regulation of leptin and the up-regulation of adiponectin. Therefore, KO could be a promising therapeutic agent for obesity by the regulation of adipokines.

The Effects of $Hwangryunhaedok$-$tang$ Pharmacopuncture by the Anti-inflammatory Action of Suppression of iNOS Production on Mice with Allergic Rhinitis (황련해독탕 약침액의 iNOS 생성 억제를 통한 항염증효과가 알레르기성 비염 유발 생쥐의 치료에 미치는 영향)

  • Cho, Jae-Yong;Kim, Yu-Jong;Kim, Eun-Jung;Lee, Seung-Deok;Kim, Kap-Sung
    • Journal of Acupuncture Research
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    • v.29 no.1
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    • pp.89-101
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    • 2012
  • Objectives : The purpose of this study was find out the therapeutic effects of its exclusive use on the rat with allergic rhinitis. Materials and Methods : Thirty Sprague-Dawley rats were divided into three group : normal group, control group and sample group. To induce the allergic rhinitis in control group and sample group, rats were sensitized intraperitoneally with 0.1% ovalumin solution 3 times at intervals of 1 week. Then intranasal sensitization was performed by diffusing 0.1% ovalumin solution 3 times at intervals of 2 days. After that time, rats in the sample group were administered by $Yonghyang$($LI_{20}$) subcutaneously to treat the inflammation. Results : 1. The anti-oxidant effects of $Hwangryunhaedok-tang$ extract was dose-dependantly increased. 2. The RAW 264.7 cells were treated with LPS for 1 hours prior to the addition of indicated concentrations ($0.4,-1.0mg/m{\ell}$) of HHT, and the cells were further incubated for 24 hours. The LPS-induced iNOS mRNA expression and NO production were dose-dependantly decreased in HHT treated RAW 264.7 cells. 3. The number of eosinophil in HP noticeably decreased than CON and this decrease had probability. The infiltration of eosinophil in HP noticeably decreased than CON. 4. The damaged mucosa as disruption of cilia in respiratory cell and vacant mucose secreting cell were increased CON, but HP same as normal configuration. Decrease of PAS positive cell were shown in CON, but goblet cell occupied with neutral mucous were shown in HP. Decrease of mucosal stress(HSP70). Decrease of perennial sign(PPAR-${\gamma}$). Decrease of icthing and sneezing intricate neurotransmitter-(substance P). 5. The anti-inflammation of HHT pharmacopuncture for AR caused mucosa comes to result as belows. Decrease of pre-inflammation cytokine(TNF-${\alpha}$). Decrease of transcription factor (NF-${\kappa}B$ p65). Decrease of transcription factor inhibitor(p-$I{\kappa}B$). Decrease of inflammation cytokine(iNOS). Conclusions : The results may suggest that administration treatment using $Hwangryunhaedok-tang$ pharmacopucnture decreases the inflammatory response on an animal model with allergic rhinitis.

Differences in Gene Expression Profiles Reflecting Differences in Drug Sensitivity to Acetaminophen in Normal and Transformed Hepatic Cell Lines In vitro

  • Jeong, Youn-Kyoung;Kang, Jin-Seok;Kim, Joo-Whan;Suh, Soo-Kyung;Lee, Michael;Kim, Seung-Hee;Lee, Sang-Kook;Park, Sue-Nie
    • Molecular & Cellular Toxicology
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    • v.5 no.1
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    • pp.32-43
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    • 2009
  • Acetaminophen (APAP) overdose is known to cause severe hepatotoxicity mainly through the depletion of glutathione. In this study, we compared the cytotoxic effects of APAP on both a normal murine hepatic cell line, BNL CL.2, and its SV40-transformed cell line, BNL SV A.8. Gene expression profiles for APAP-treated cells were also obtained using microarray and analyzed to identify differences in genes or profiles that may explain the differences of susceptibility to APAP in these cell lines. These two cell lines exhibited different susceptibilities to APAP (0-$5,000{\mu}M$); BNL SV A.8 cells were more susceptible to APAP treatment compared to BNL CL.2 cells. A dose of $625{\mu}M$ APAP, which produced significant differences in cytotoxicity in these cell lines, was tested. Microarray analysis was performed to identify significant differentially expressed genes (DEGs) irrespective of APAP treatment. Genes up-regulated in BNL SV A.8 cells were associated with immune response, defense response, and apoptosis, while down-regulated genes were associated with catalytic activity, cell adhesion and the cytochrome P450 family. Consistent with the cytotoxicity data, no significant DEGs were found in BNL CL.2 cells after treatment with $625{\mu}M$ APAP, while cell cycle arrest and apoptosis-related genes were up-regulated in BNL SV A.8 cells. Based on the significant fold-changes in their expression, a genes were selected and their expressions were confirmed by quantitative real-time RT-PCR; there was a high correlation between them. These results suggest that gene expression profiles may provide a useful method for evaluating drug sensitivity of cell lines and eliciting the underlying molecular mechanism. We further compared the genes identified from our current in vitro studies to the genes previously identified in our lab as regulated by APAP in both C57BL/6 and ICR mice in vivo. We found that a few genes are regulated in a similar pattern both in vivo and in vitro. These genes might be useful to develop as in vitro biomarkers for predicting in vivo hepatotoxicity. Based on our results, we suggest that gene expression profiles may provide useful information for elucidating the underlying molecular mechanisms of drug susceptibility and for evaluating drug sensitivity in vitro for extrapolation to in vivo.