• Title/Summary/Keyword: ST cell

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miR-375 down-regulation of the rearranged L-myc fusion and hypoxia-induced gene domain protein 1A genes and effects on Sertoli cell proliferation

  • Guo, Jia;Liu, Xin;Yang, Yuwei;Liang, Mengdi;Bai, Chunyan;Zhao, Zhihui;Sun, Boxing
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.8
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    • pp.1103-1109
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    • 2018
  • Objective: This study aimed to screen and identify the target genes of miR-375 in pig Sertoli (ST) cells and to elucidate the effect of miR-375 on the proliferation of ST cells. Methods: In this study, bioinformatics software was used to predict and verify miR-375 target genes. Quantitative polymerase chain reaction (PCR) was used to detect the relationship between miR-375 and its target genes in ST cells. Enzyme-linked immunosorbent assay (ELISA) of rearranged L-myc fusion (RLF) and hypoxia-induced gene domain protein 1A (HIGD1A) was performed on porcine ST cells, which were transfected with a miR-375 mimics and inhibitor to verify the results. Dual luciferase reporter gene assays were performed to assess the interactions among miR-375, RLF, and HIGD1A. The effect of miR-375 on the proliferation of ST cells was analyzed by CellTiter 96 AQueous One Solution Cell Proliferation Assay (MTS). Results: Five possible target genes of miR-375, including RLF, HIGD1A, colorectal cancer associated 2, POU class 3 homeobox 1, and WW domain binding protein 1 like, were found. The results of quantitative PCR suggested that mRNA expression of RLF and HIGD1A had a negative correlation with miR-375, indicating that RLF and HIGD1A are likely the target genes of miR-375. The ELISA results revealed that RLF and HIGD1A were negatively correlated with the miR-375 protein level. The luminescence results for the miR-375 group cotransfected with wild-type RLF and HIGD1A vector were significantly lower than those of the miR-375 group co-transfected with the blank vector or mutant RLF and HIGD1A vectors. The present findings suggest that RLF and HIGD1A are target genes of miR-375 and that miR-375 inhibits ST cell proliferation according to MTS analysis. Conclusion: It was speculated that miR-375 affects cell proliferation through its target genes, which play an important role in the development of testicular tissue.

The Change of Taurine Transport in Osteocytes by Oxidative Stress, Hypertonicity and Calcium Channel Blockers

  • Kang, Young-Sook;Kim, Soon-Joo
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.219-225
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    • 2008
  • Taurine is the most abundant amino acid in many tissues and is found to be enhancing the bone tissue formation or inhibits the bone loss. Although it is reported that taurine reduces the alveolar bone loss through inhibiting the bone resorption, its functions of taurine and expression of taurine transporter (TauT) in bone have not been identified yet. The purpose of this study is to clarify the uptake mechanism of taurine in osteoblast using mouse osteoblast cell lines. In this study, mouse stromal ST2 cells and mouse osteoblast-like MC3T3-E1 cells as osteoblast cell lines were used. The activity of taurine uptake was assessed by measuring the uptake of [$^3H$]taurine in the presence or absence of inhibitors. TauT mRNA was detected in ST2 and MC3T3-E1 cells. [$^3H$]Taurine uptake by these cells was dependent on the presence of extracellular calcium ion. The [$^3H$]taurine uptake in ST2 cells treated with 4 mM calcium was increased by 1.7-fold of the control which was a significant change. In contrast, in $Ca^{++}$-free condition and L-type calcium channel blockers (CCBs), taurine transport to osteocyte was significantly inhibited. In oxidative stress conditions, [$^3H$]taurine uptake was decreased by TNF-$\alpha$ and $H_2O_2$. Under the hyperosmotic conditions, taurine uptake was increased, but inhibited by CCBs in hyperosmotic condition. These results suggest that, in mouse osteoblast cell lines, taurine uptake by TauT was increased by the presence of extracellular calcium, whereas decreased by CCBs and oxidative stresses, such as TNF-$\alpha$ and $H_2O_2$.

Study of Experimentations and Clinical Trials' Trends for Obesity Treatment using Pharmacupuncture (약침을 활용한 비만연구의 실험실적 및 임상적 연구동향 고찰)

  • Kim, Min-Woo;Song, Yun-Kyung;Lim, Hyung-Ho
    • Journal of Korean Medicine for Obesity Research
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    • v.11 no.1
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    • pp.47-60
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    • 2011
  • Objectives The objective of this study was to contribute to the development of pharmacupuncture for obesity treatment by reviewing the studies of pharmacupuncture experimentations and clinical trials. Methods We searched the papers with keywords of 'obesity' and 'pharmacupuncture' in the search site, RISS, Oriental medicine advanced searching integrated system(OASIS), Korean traditional knowledge portal, the society of Korean medicine for obesity research, Korean pharmacupuncture institute, the Korean academy of oriental rehabitation medicine and Korean acupuncture & moxibustion society. Results 1. We reviewed 37 articles searched. 27 articles(73.0%) were animal experimentations, 5 articles(13.5%) were cell experimentations, 4 articles(10.8%) were clinical trials and 1 article(2.7%) was study analysis. 2. The herbs, using for animal experimentations, were atratylodes japonica, coix lachrymajobi, ephedra sinica, crataegus pinnatifida, wild ginseng and etc. Acupucture points were joksamni(ST36), zhongwan(CV12), gansoo(BL18), pungnyung(ST40), umnungchon(SP9), bisu(BL20), gokji (LI11), cheun-chu(ST25) and etc. 3. For cell experimentations, preadipocytes and adipocytes performed on cell cultures with using rats, 3T3-L1 preadipocytes and porcine skin including fat tissue were treated with fel ursi, bovis calculus, ephedrae herba, spirodelae herba, wild ginseng. 4. For clinical trials, Sangsik no.1, Bigiheo, ephedra, green tea and sweet bee venom were injected at the region where a lot of fat like zhongwan(CV12), xiawan(CV10), kwanwon(CV4), cheun-chu(ST25) and thigh. Conclusion Through animal and cell experimentations and clinical trials, the treatment of obesity using local acupuncture therapy was effective. For clinical use, however, it is considered that animal and cell experimentation and clinical trial's connection using one kind of herb and studies about more clinical trials and associated side effects are needed.

Engineering Human-like Sialylation in CHO Cells Producing hCTLA4-Ig by Overexpressing α2,6-Sialyltransferase (α2,6-Sialyltransferase 과발현을 통한 인간형 시알산 부가 hCTLA4-Ig 생산 CHO 세포주 제작)

  • Lim, Jin-Hyuk;Cha, Hyun-Myoung;Park, Heajin;Kim, Ha Hyung;Kim, Dong-Il
    • KSBB Journal
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    • v.32 no.3
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    • pp.193-198
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    • 2017
  • Sialylation is important in producing therapeutic proteins such as antibody, cytokine and fusion protein. Thus, enhancement of sialylation is usually performed in CHO cell cultures. ${\alpha}2,6$-Sialyltransferase (ST), which plays a key role in the attachment of ${\alpha}2,6-sialic$ acid, is present in human cells but not in Chinese hamster ovary (CHO) cells. Overexpression of ${\alpha}2,6-ST$ can be used for enhancing the degree of sialylation and achieving human-like glycosylation. In this study, we constructed CHO cells producing human cytotoxic T-lymphocyte antigen4-immunoglobulin (hCTLA4-Ig) as well as ${\alpha}2,6-ST$. Transfected CHO cells were selected using G418 and stable cell line was established. Profiles of viable cell density and hCTLA4-Ig titer in an overexpressed cell line were similar to those of a wild-type cell line. It was confirmed that the total amount of sialic acid was increased and ${\alpha}2,6-sialic$ acid was attached to the terminal residues of N-glycan of hCTLA4-Ig by ESI-LC-MS. Compared to 100% of ${\alpha}2,3-sialic$ acid in wild type cells, 70.9% of total sialylated N-glycans were composed of ${\alpha}2,6-sialic$ acid in transfected cells. In conclusion, overexpression of ${\alpha}2,6-ST$ in CHO cells led to the increase of both the amount of total sialylated N-glycan and the content of ${\alpha}2,6-sialic$ acid, which is more resemble to human-like structure of glycosylation.

Pentoxifylline Induces Lipolysis and Apoptosis of Human Preadipocytes, Keratinocytes and Fibroblasts In Vitro

  • Lee, Il-Kyu;Choi, Yun-Jung;Shim, In-Sop;Kim, Kyung-Soo;Choi, Chang-Jin
    • Biomolecules & Therapeutics
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    • v.18 no.1
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    • pp.56-64
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    • 2010
  • Pentoxifylline (PTX) has been used for the local reduction of fat tissue in the clinical setting. However, its safety and efficacy have not been proven. The aim of this study was to evaluate the effects of PTX on cell lines established from fat tissue. Newly cultured human preadipocytes and adipocytes from subcutaneous abdominal fat in addition to purchased human lung fibroblasts and keratinocytes were treated with PTX at different concentrations. Cell viability was determined using the Cell counting kit (CCK)-8 assay and lipolysis was evaluated using an Elisa kit. DNA fragmentation, Western blot analysis, Hoechst and Propidium Iodide (PI) staining and fluorescence activated cell scanning analysis were performed to confirm apoptosis. The viability of adipocytes, preadipocytes, keratinocytes and fibroblasts was markedly decreased at concentrations of PTX above 20 mM. Apoptosis was induced at concentrations of PTX over 40 mM in all cell lines. Lipolysis was increased by 60% at concentrations of PTX of 20 mM compared to the control. In conclusion, the results of this study showed that 20 mM of PTX induced lipolysis. At concentrations over 20 mM, PTX reduced the viability of all cells studied including: adipocytes, preadipocytes, fibroblasts and keratinocytes, in a non-specific manner.

Attenuation of Hepatic Graft-versus-host Disease in Allogeneic Recipients of MyD88-deficient Donor Bone Marrow

  • Lim, Ji-Young;Lee, Young-Kwan;Lee, Sung-Eun;Ju, Ji-Min;Park, Gyeongsin;Choi, Eun Young;Min, Chang-Ki
    • IMMUNE NETWORK
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    • v.15 no.3
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    • pp.125-134
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    • 2015
  • Acute graft-versus-host-disease (GVHD) is characterized by selective damage to the liver, the skin, and the gastrointestinal tract. Following allogeneic hematopoietic stem cell transplantation, donor bone marrow (BM) cells repopulate the immune system of the recipient. We previously demonstrated that the acute intestinal GVHD (iGVHD) mortality rate was higher in MyD88-deficient BM recipients than that in the control BM recipients. In the present study, the role of MyD88 (expressed by donor BM) in the pathophysiology of hepatic GVHD (hGVHD) was examined. Unlike iGVHD, transplantation with MyD88-deficient T-cell depleted (TCD) BM attenuated hGVHD severity and was associated with low infiltration of T cells into the liver of the recipients. Moreover, GVHD hosts, transplanted with MyD88-deficient TCD BM, exhibited markedly reduced expansion of $CD11b^+Gr-1^+$ myeloidderived suppressor cells (MDSC) in the liver. Adoptive injection of the MDSC from wild type mice, but not MyD88-deficient mice, enhanced hepatic T cell infiltration in the MyD88-deficient TCD BM recipients. Pre-treatment of BM donors with LPS increased MDSC levels in the liver of allogeneic wild type BM recipients. In conclusion, hGVHD and iGVHD may occur through various mechanisms based on the presence of MyD88 in the non-T cell compartment of the allograft.

Bilateral Localized Tenosynovial Giant Cell Tumor Causing Patellofemoral Pain Syndrome (슬개대퇴 통증 증후군을 초래한 양측성 건활막 거대세포종)

  • Chung, Whan-Yong;Shin, Sung-Chul;Yi, Sang-Yeop;Sung, Seung-Yong
    • Journal of the Korean Orthopaedic Association
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    • v.56 no.6
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    • pp.525-529
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    • 2021
  • A localized tenosynovial giant cell tumor rarely affects the bilateral knee joint compared to the diffuse form. Only a few cases can be found in the literature, and there is no case report of the bilateral localized form of giant cell tumor causing patellofemoral pain syndrome in Korea. This study experienced a case of bilateral localized giant cell tumor, causing patellofemoral pain syndrome. The tumor was excised arthroscopically and confirmed pathologically. This paper reports this case with a review of the relevant literature.

Prognostic Factors in Stage IIB Non-Small Cell Lung Cancer according to the 8th Edition of TNM Staging System

  • Shin, Jin Won;Cho, Deog Gon;Choi, Si Young;Park, Jae Kil;Lee, Kyo Young;Moon, Youngkyu
    • Journal of Chest Surgery
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    • v.52 no.3
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    • pp.131-140
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    • 2019
  • Background: The purposes of this study were to evaluate the appropriateness of the stage migration of stage IIA non-small cell lung cancer (NSCLC) in the seventh edition of the tumor, node, and metastasis classification for lung cancer to stage IIB lung cancer in the eighth edition, and to identify prognostic factors in patients with eighth-edition stage IIB disease. Methods: Patients with eighth-edition stage IIB disease were subclassified into those with seventh-edition stage IIA disease and those with seventh-edition stage IIB disease, and their recurrence-free survival and disease-specific survival rates were compared. Risk factors for recurrence after curative resection were identified in all included patients. Results: Of 122 patients with eighth-edition stage IIB NSCLC, 101 (82.8%) had seventh-edition stage IIA disease and 21 (17.2%) had seventh-edition stage IIB disease. Nonsignificant differences were observed in the 5-year recurrence-free survival rate and the 5-year disease-specific survival rate between the patients with seventh-edition stage IIA disease and those with seventh-edition stage IIB disease. Visceral pleural invasion was a significant risk factor for recurrence in patients with eighth-edition stage IIB NSCLC. Conclusion: The stage migration from seventh-edition stage IIA NSCLC to eighth-edition stage IIB NSCLC was appropriate in terms of oncological outcomes. Visceral pleural invasion was the only prognostic factor in patients with eighth-edition stage IIB NSCLC.

A Study on Anti-aning of Electroacupuncture at Joksamni($ST_{36}$) in Rats (족삼리(足三里) 전침(電鍼)이 흰쥐의 노화(老化)에 미치는 영향(影響))

  • Lee, Jun-Seong;Yim, Yun-Kyoung;Kim, Young-Il
    • Journal of Acupuncture Research
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    • v.24 no.3
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    • pp.145-161
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    • 2007
  • Objectives : The purpose of this study is to investigate the effect of electroacupuncture at Joksanmi($ST_{36}$) on Rat's aging by experimental methods. Methods : The author performed several experimental procedures to observe the effects of the EC-HAS at the arthritis. First, I measured the cell survival rate of the mice lung fibroblasts. Second, the incidence rate of arthritis and arthritis index of CIA were analyzed. Third, the levels of IL-6, $INF-{\gamma}$ , $INF-{\alpha}$ , $IL-1{\beta}$, IgG, IgM and anti-collagen II were measured in serum and the level of $INF-{\gamma}$, $INF-{\gamma}$/IL-4 ratio in the CIA mouse spleen cell culture. Fourth, histological analysis of the mice joint was performed. Fifth, the expression ratio of $CD3e^+$ cells to $CD19^+$ cells, $CD4^+$ cells to $CD8^+$ cells, $CD11a^+CD19^+$ cells, $CD4^+CD25^+$ cells, $CD3^+CD69^+$ cells, and $CD11b^+Gr-1^+$ cells were checked. Results: 1. The hematocrit was meaningfully (p<0.01, p<0.05) decreased in $ST_{36}$ group in comparison with the control and sham group. 2. The glucose, ALP were meaningfully (p<0.05, p<0.05) decreased in $ST_{36}$ group in comparison with the control group. 3. In the activation of hepatic antioxidase, the catalase was meaningfully (p<0.01) increased in $ST_{36}$ group in comparison with the control group. 4. In the liver, the NO formation was meaningfully (p<0.001) decreased in $ST_{36}$ group in comparison with the nomal, control and sham group. 5. In the activation of splenetic antioxidase, the GSH was meaningfully (p<0.01) increased in $ST_{36}$ group in comparison with the sham group, and the catalase was meaningfully (p<0.001) increased in $ST_{36}$ group in comparison with the control group. 6. In the spleen, the MDA formation was meaningfully (p<0.01, p<0.05) decreased in $ST_{36}$ group in comparison with the control and sham group. 7. In the spleen cell culture, the levels of IL-6 was meaningfully (p<0.05) decreased in comparison with the sham group, $INF-{\gamma}$( was meaningfully (p<0.05) increased in comparison with the control group. Conclusions: According to the results, it is considered that electroacupuncture at Joksamni($ST_{36}$) is effective in increasing the activities of antioxidative enzyme and inhibiting lipid peroxides, and has an immunomodulatory reaction. In conclusion, it han an significant anti-aging effect and need more experimental study.

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