Purpose: The objective of this study was conducted to investigate the effects of rutin, buckwheat components on cell growth and anti-inflammation in adipocyte 3T3-L1 and human colon cancer cell SW-480. Methods: We cultured 3T3-L1 adipocyte and SW-480 colon cancer cell to confluence, at which time starvation was induced with SFM for 1 day. Cells were then cultured in medium containing 0, 25, 50, or 100 μmol/mL of rutin 3T3-L1 or 0, 10, 20, or 40 μmol/mL SW-480. Cell viability was measured using a cell viability kit. In addition, we examined the expression of mRNA related to inflammation. RT-PCR was used to quantity tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), IL-6, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) mRNA levels. Results: Rutin significantly inhibited 3T3-L1 and SW-480 cell proliferation in a dose and time dependent manner. Rutin also significantly reduced the mRNA expression of IL-1β, IL-6 and TNF-α at the highest dose. In addition, rutin treatment caused a significant reduction in COX-2 and iNOS mRNA levels compared to the control group. Conclusion: Overall, our results suggest that rutin has the potential to reduce inflammation, and that these effects are greater during tissue-damaging inflammatory conditions.

The knock-in efficiency is very important to manipulate gene editing in the transgenic domestic animal. Recently, it is reported that transiently loosen nucleosome folding of transcriptionally inactive chromatin might have potential tp enhance the homologouse recombination efficiency. Histone deacetylases (HDAC) are a class of enzymes that remove acetyl groups from an amino acid on a histone. This is important because DNA is wrapped around histones, and DNA expression is regulated by acetylation and de-acetylation. In this study, Mac-T cell were treated with 10uM VPA (valproic acid, HDAC inhibitor) for 24 h and transfected with Knock-in vector and TALEN at targeting of β-casein gene. After 3 day of transfection, knock-in efficiency was confirmed by PCR. The level of HDAC2 protein in Mac-T cells was decreased by VPA treatment. The knock-in efficiency in the Mac-T cell with treated HDAC inhibitor was higher than cell not treated HDAC inhibitor. These results indicated that chromatin modification by HDAC inhibitor enhances homologous recombination efficiency in the Mac-T cell.

Purpose: This study examined the effects of α-lipoic acid in diluted solvents on cell growth in 3T3-L1 cells according to the treated concentration and times. Methods: Adipocyte 3T3-L1 cell were cultured. Confluent cells underwent starvation with SFM for 1 day and then were cultured in a medium containing various concentrations 0, 100, 200, and 400 μmol/L of α-lipoic acid. The cell viability was measured using the EZ Cytox assay kit. In addition, the effect of α-lipoic acid of diluted solvents on the cell growth in 3T3-L1cells was examined according to the treated concentration and times. Results: The α- lipoic acid diluted ethanol inhibited cell proliferation in a dose and time dependent manner. The α-lipoic acid diluted ethanol induced adipocyte 3T3-L1 cells proliferation with an adipocyte inducer. In addition, α-lipoic acid inhibited adipocyte 3T3- L1 growth in a dose and time dependent manner (p<0.05). Conclusion: This study showed that a treatment with α-lipoic acid diluted ethanol inhibits cell growth of, adipocyte 3T3-L1 cells induced with an adipocyte inducer, (200 μmol/L of α- lipoic acid) treated for 48 hr.

Pleurotus cornucopiae (PC) mushrooms is found in the field and commonly known in Japan as Tamogidake mushrooms. Recently it has been reported that PC also alleviating the toxicity of heavy metals. However little is known about mechanism of the action of PC on osteoblast differentiation, especially in transcription factor. Inhibitor of DNA binding-1 (Id-1) function has been linked to the proliferation, migration, and senescence of cells, and studies about relationship between Id-1 and biological function. Therefore, this study was aimed to investigate the effect of PC on osteoblast differentiation and expression of Id-1 and Id-2. PC treatment increased ALP, Col 1 and OCN. PC treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This PC–induced osteoblast differentiation is more effective in lower doses rather than high doses. This study shows that expression of Id-1 and Id-2 was increased in a dose-dependent manner during PC-induced osteoblast differentiation.

We would like to report a case of bovine lymphosarcoma. Parous cattle from a livestock farmhouse were examined for mutinodular masses in the abdominal cavity after slaughter. For clinical signs, animals presented mild leukemic signs but did not have viral or bacterial infection. Grossly, whitish to yellowish smooth masses similar to fat tissue were covered with a thin membrane. A multilobulated mass formed around the arteri, and there was a large quantity of reddish fluid on the cut surface. Histopathologically, a monomor- phic population of lymphocytes was observed along with small amounts of cytoplasm, round nuclei with coarsely granular chromatin, and numerous mitotic figures in the samples. In the tumor lesion, uniformly round cells had invaded with abundant neovascularization. Especially, the immunohistochemical phenotype of tumor cells was posi- tive for anti-CD3 and negative for anti-CD8 and anti-CD20. Therefore, morphological analysis diagnosed the mass as a multinodular bovine lymphosarcoma of T-cell origin with- out any sign of infection by a viral agent.

The T-cell receptor (TCR) engages with an antigen and initiates a signaling cascade that leads to the activation of transcription factors. Roquin, a protein encoded by the RC- 3H1 gene and characterized as an immune regulator, was recently identified as a novel RING-type ubiquitin ligase family member, but the mechanisms by which Roquin regulates T-cell responses are unclear. We used the EL-4 murine lymphoma cell line to elucidate the role of Roquin in vitro. Roquin-overexpressing EL-4 cells became hyper-responsive after anti-CD3/CD28 stimulation in vitro and were a major source of the cytokines IL-2 and TNF-α. Upon activation, these cells showed particularly enhanced production of IL-2 and TNF-α. To clarify the important role played by Roquin in T-cell responses ex vivo, we generated T-cell-specific Roquin transgenic (Tg) mice. Roquin-Tg CD4+ T-cells showed enhanced production of IL-2 and TNF-α in response to TCR stimulation with anti-CD28 co-stimulation. Further studies are necessary to investigate the role of Roquin in the regulation of primary T-cell activation, survival, and differentiation.

MAC-T cells, bovine mammary epithelial cell line, have been utilized to investigate bovine lactation system. A lactogenic phenotype of the cell is generally induced by combination of dexamethasone, insulin and prolactin (PRL). Effect of vitamin A derivative retinoic acid (RA), well reported as an inducer for differentiation in many cells, to MAC-T cell has not been studied. The objective of this study was to confirm effect of differentiation potential by RA treatment in MAC-T cells and to test effect of combination of RA and PRL treatment. In RA or PRL treatment groups, both has induced morphological change to secrete milk of MAC-T cells. Combination of RA and PRL treatment group has presented noticeable lactogenic phenotype among the all group. This phenotype observed at four days after treatment and showed critical morphological change that was rouphly spherical structure at eight days. RA alone treatment showed slightly inhibition of proliferation in the MAC-T cells, but co-treatment with PRL was improved the cell growth more than control group. MTT assay result and Bcl-xL/Bax ratio of mRNA abundance also was entirely consistent with earlier one. RA-induced differentiation of MAC-T cells has increased αs1-casein, αs2-casein and β-casein mRNA expression compared to PRL treatment group. Expression of αs1-casein, αs2-casein and β-casein genes represented the maximum value in the combination of RA and PRL treatment group at four days. The value of each casein gene expression was 4-, 5.5- and 5.9-fold, respectively, as compared with PRL alone treatment in the MAC-T cells. Protein level of β-casein releasing to the medium also induced the highest level at four days. These results provide evidence that RA can induce the differentiation of MAC-T cells and have synergetic effect with PRL.

Several types of white blood cells, such as T cells, B cells, and macrophages, are involved in the immune response. In particular, the processes of T-cell activation play a crucial role in an adaptive immune response, whereby the T-cell receptor (TCR) engages with an antigen and signals a cascade that leads to the activation of transcription factors (AP-1, NF-κB, and NFAT) that are critically involved in cytokine production. Roquin, encoded by the RC3H1 gene and characterized as an immune regulator, was recently identified as a novel RING-type ubiquitin ligase family member, but the mechanisms by which Roquin proteins regulate T-cell responses are unclear. To elucidate the role of Roquin in vitro, murine lymphoma EL-4 cells were used. Roquin overexpressing Tcells became hyper-responsive upon anti-CD3/CD28 stimulation in vitro and were a major source of cytokines such as IL-2, TNF-α, IL-6, and IL-10. Upon activation, these cells showed preferentially enhanced production of IL-2 and TNF-α, but not IFN-γ. To clarify the important role of Roquin in the T-cell response ex vivo, we generated T-cell-specific Roquin-transgenic (Tg) mice having a higher expression of Roquin in T cells as compared to wild-type mice. Using Roquin-Tg mice, we studied whether immune responses are affected ex vivo. Roquin-Tg CD4+ T cells showed enhanced production of IL-2 or TNF-α to TCR stimulation with anti-CD28 costimulation via up-regulation of CD28. T-cell proliferation also increased in Roquin-Tg CD4+ T cells after anti-CD3/CD28 treatment. Further studies on the role of Roquin in the regulation of primary T-cell activation, survival, and differentiation may be anticipated.

Facial numbness restricted to the distribution of the mental nerve(mental neuropathy) is called "numb chin syndrome". The clinical importance of this syndrome is associated with first recognition of involvement of malignant diseases. The malignant neoplasm with numb chin syndrome show rapid progression and high mortality. We present a 43-year-old female who had been treated by radiotherapy for precursor T-cell leukemia/lymphoma involving the central nervous system( CNS) previously and later developed mental nerve invasion without central nervous system recurrence. MRI images revealed the CNS tumor mass remitted, and there was no identified peripheral nervous system(PNS) involvement including the mental nerve invasion, nevertheless the patient complained of consistent numbness and pain on right mandibular area. This is the first case of precursor T-cell leukemia/lymphoma accompanying mental nerve invasion without recurrence for central nervous system. Proper interpretation for mental neuropathy may lead to the prompt diagnosis and therapeutic intervention.