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extracelluar RNA

Luanluan Shao, Chaochao Xu, Hongshuai Ji, Weiping Mao, Yingying Wang, Xiaoqian Liu, Yanyan Zhu
Objective To construct the ribosome display library of anti-B7-H4 extracellular domain, and select the antibody with high specificity. Methods The cDNA of B7-H4 extracellular domain was amplified from A549 cells by reverse transcription PCR (RT-PCR). To express ectodomains of B7-H4, the sequence of B7-H4 gene, which encodes the B7-H4 extracellular domains, was inserted into plasmid pET-28a(+). The purified recombinant protein of B7-H4 extracellular domain was used to immunize BALB/c mice. The total RNA was extracted from the spleen of BALB/c mice which had been immunized with B7-H4 recombinant protein...
September 2016: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
Randy J Hempel, John P Bannantine, Judith R Stabel
Johne's disease is a chronic infection of the small intestine caused by Mycobacterium avium subspecies paratuberculosis (MAP), an intracellular bacterium. The events of pathogen survival within the host cell(s), chronic inflammation and the progression from asymptomatic subclinical stage to an advanced clinical stage of infection, are poorly understood. This study examines gene expression in the ileocecal valve (ICV) of Holstein dairy cows at different stages of MAP infection. The ICV is known to be a primary site of MAP colonization and provides an ideal location to identify genes that are relevant to the progression of this disease...
2016: PloS One
Erika H Noss, Gerald F M Watts, Davide Zocco, Tracy L Keller, Malcolm Whitman, Carl P Blobel, David M Lee, Michael B Brenner
INTRODUCTION: Engagement of the homotypic cell-to-cell adhesion molecule cadherin-11 on rheumatoid arthritis (RA) synovial fibroblasts with a chimeric molecule containing the cadherin-11 extracellular binding domain stimulated cytokine, chemokine, and matrix metalloproteinases (MMP) release, implicating cadherin-11 signaling in RA pathogenesis. The objective of this study was to determine if cadherin-11 extracellular domain fragments are found inside the joint and if a physiologic synovial fibroblast cleavage pathway releases those fragments...
2015: Arthritis Research & Therapy
Xiaolin Wu, Hong Jiang, Jing Chen, Qi Hu, Jing Zhang, Jichun Wang
OBJECTIVE: To explore the effects of Kindlin-2 RNA interference on vascular smooth muscle cells (VSMCs) migration, adhesion and β1-integrin as well as the relationship between Kindlin-2 and β1-integrin. METHODS: Primary VSMCs were cultured, infected with Kindlin-2 siRNA lentiviral vectors.VSMCs were divided into three groups:the blank control group, the negative control group and the Kindlin-2 siRNA group. The ability of VSMCs migration was measured by Transwell experiment and wound healing assay...
November 2014: Zhonghua Xin Xue Guan Bing za Zhi
Biswaprakash Pradhan, Sashi K Dash, Sabuj Sahoo
OBJECTIVE: To screen and isolate an eco-friendly, a thermophilic and potent L-asparaginase producing bacterium, with novel immunological properties that may obviates hypersensitivity reactions. METHODS: In the present study bacterial strain isolated for extracellular L-asparaginase production from hotspring, identified by morphological, biochemical and physiological tests followed by 16S rDNA technology and the L-asparaginase production ability was tested by both semi quantitative and quantitative enzymatic assay...
December 2013: Asian Pacific Journal of Tropical Biomedicine
Akihiro Ishii, Sharyl L Fyffe-Maricich, Miki Furusho, Robert H Miller, Rashmi Bansal
Wrapping of the myelin sheath around axons by oligodendrocytes is critical for the rapid conduction of electrical signals required for the normal functioning of the CNS. Myelination is a multistep process where oligodendrocytes progress through a well coordinated differentiation program regulated by multiple extracellular growth and differentiation signals. The intracellular transduction of the extracellular signals that regulate myelination is poorly understood. Here we demonstrate a critical role for two important signaling molecules, extracelluar signal-regulated protein kinases 1 and 2 (ERK1/ERK2), downstream mediators of mitogen-activated protein kinases, in the control of CNS myelin thickness...
June 27, 2012: Journal of Neuroscience: the Official Journal of the Society for Neuroscience
Ping Li, Jianxin Qian, Guanzhen Yu, Ying Chen, Ke Liu, Jie Li, Jiejun Wang
BACKGROUND: SPARC-like protein 1 (SPARCL1), a member of extracelluar matrix glycoprotein, is involved in many physiological functions. METHODS: Tissue microarray (TMA) blocks were constructed based on 1,072 Chinese patients, containing both gastric cancer (GC) tissues and adjacent normal mucosa tissues. We analyzed the expression of SPARCL1 from both mRNA and protein level, using Real-time quantitative polymerase chain reaction (qRT-PCR), semi-quantitative PCR, immunohistochemistry (IHC), and Western blotting...
January 2012: Journal of Surgical Oncology
Huirong Chen, Lixian Fu, Lingxi Luo, Jun Lu, W Lindsey White, Zhangli Hu
The viable but nonculturable (VBNC) state has been found to be a growth strategy used by many aquatic pathogens; however, few studies have focused on VBNC state on other aquatic bacterial groups. The purpose of this study was to explore the VBNC state of cyanobacteria-lysing bacteria and the conditions that regulate their VBNC state transformation. Three cyanobacteria-lysing heterotrophic bacterial strains (F1, F2 and F3) were isolated with liquid infection method from a lake that has experienced a cyanobacterial bloom...
January 2012: Microbial Ecology
Jing Bai, Xiaoning Zhong, Haijuan Tang, Zhiyi He, Jianquan Zhang, Jingming Deng
BACKGROUND AND OBJECTIVE: Recent studies have shown that integrin α5β1 as a core in the integrin family plays an important role in metastasis, invasion and poor difference of non-small cell lung cancer. In this study, A549 cells were cultured and treated with integrin α5β1 small interfering RNA (siRNA) and extracelluar signal-regulated protein kinase (ERK) inhibitor PD98059 to investigate the effect of integrin α5β1 on proliferation and migration of A549 cells and explore its signal transduction mechanism...
July 2011: Zhongguo Fei Ai za Zhi, Chinese Journal of Lung Cancer
Chan Zhu, Yuqin Pan, Bangshun He, Bo Wang, Yongfei Xu, Lili Qu, Qian Bao, Fuliang Tian, Shukui Wang
CD147, also named extracelluar matrix metalloproteinase inducer (EMMPRIN), is a member of the immunoglobulin family and a glycoprotein enriched on the surface of tumor cells, which promotes invasion, metastasis, growth and survival of malignant cells, and is known to confer resistance to some chemotherapeutic drugs. To determine the possible role of CD147 in the invasive properties of laryngeal carcinoma, we used an RNA interference approach to silence CD147 expression in the Hep2 cell line at high levels of CD147 expression...
February 2011: Oncology Reports
Cindy M Sondag, Colin K Combs
BACKGROUND: Amyloid precursor protein (APP) is a ubiquitously expressed cell surface protein reported to be involved in mediating cell-cell or cell-matrix interactions. Prior work has demonstrated that APP co-localizes with beta1 integrin in different cell types. METHODS: In an effort to determine the function of APP on monocytic lineage cells, in particular, the human monocyte cell line, THP-1, was used to assess the role of APP during adhesion to the extracelluar matrix component type I collagen...
March 19, 2010: Journal of Neuroinflammation
Hai Dan Yuan, Sung Hyun Chung
In this study, we investigated the protective effects of fermented ginseng (FG) on hyperglycemia induced by streptozotocin (STZ) in Sprague Dawley rats. FG was administered orally at dose of 250 (FGL) or 500 mg/kg (FGH) for 20 days starting one week before STZ injection. FG restored the plasma insulin levels by 266% and 334% in FGL and FGH, respectively, and resulting in reduction of plasma glucose concentration. Histological observation indicated that STZ-induced destruction of pancreatic islets was protected by FG...
June 2010: Phytotherapy Research: PTR
Wei Xiong, Peng-yan Niu, Wen-tao Zhu, Jing Chen
BACKGROUND: Treating metastatic osteosarcoma has been challenged in past decades. Extracelluar matrix (ECM) proteins play an important role in the progression of osteosarcoma as they are pivotal components of the tumor microenvironment. Here, we identified potential genes belonging to the ECM and characterized the roles of these genes in the progression of osteosarcoma and their association with outcomes. METHODS: Osteosarcoma parental cell line MG63 and its derivative MG63-A1 with a high metastatic potential underwent oligonucleotide microarray analysis...
November 20, 2009: Chinese Medical Journal
Jinping Luo, Susan Megee, Ina Dobrinski
Asymmetric division of germline stem cells in vertebrates was proposed a century ago; however, direct evidence for asymmetric division of mammalian spermatogonial stem cells (SSCs) has been scarce. Here, we report that ubiquitin carboxy-terminal hydrolase 1 (UCH-L1) is expressed in type A (A(s), A(pr), and A(al)) spermatogonia located at the basement membrane (BM) of seminiferous tubules at high and low levels, but not in differentiated germ cells distant from the BM. Asymmetric segregation of UCH-L1 was associated with self-renewal versus differentiation divisions of SSCs as defined by co-localization of UCH-L1(high) and PLZF, a known determinant of undifferentiated SSCs, versus co-localization of UCH-L1(low/-) with proteins expressed during SSC differentiation (DAZL, DDX4, c-KIT)...
August 2009: Journal of Cellular Physiology
Dongming Wang, Linmei Shi
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a new member of the TNF superfamily. In this paper, we report the expression, purification, and preparation of a recombinant form of the extracelluar domain of the TRAIL (sTRAIL) without posttranslational modifications, which may selectively induce apoptosis of tumor cells in vitro. To obtain recombinant nonfusion sTRAIL protein, the encoding region for sTRAIL was cloned between KpnI and BamHI in pET32a. The Trx (thioredoxin)/sTRAIL fusion proteins were expressed in the form of inclusion bodies in Escherichia coli host strain BL21 (DE3)...
April 2009: Applied Biochemistry and Biotechnology
Jun Li, Xiaoping Yi, Yuanxing Zhang, Xiangming Sun
The extracelluar domain I-IV of target gene VEGFR2 (Vascular endothelial growth factor receptor 2) was cloned from villus of trimester abortion by RT-PCR, and linked to the expression vectors. Then, the transfection conditions were optimized in serum-free suspension culture HEK293 using GFP (Green fluorescence protein) as the report gene. The results showed that the optimal transfection efficiency and cell number were obtained when the ratio of foreign DNA: PEI = 1:2 (W/W), DNA = 1.5 g /10(6) cells and shaking speed (120 r/min) in serum free medium in the beginning 4 hours of transfection...
May 2008: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Xiang Chen, Juan Su, Jing Chang, Takuro Kanekura, Ji Li, Ye Hong Kuang, Sheng Peng, Fang Yang, Hui Lu, Jiang Lin Zhang
CD147, a leukocyte surface molecule over-expressed in T-lymphoma cells, is reportedly associated with lymphocyte activation and proteinase production via interactions with fibroblasts and plays a role in stromal invasion by lymphoma cells. To determine the role of CD147 in the progression of T-lymphoma, we performed siRNA interference-mediated knockdown of CD147 in a CD147-expressing Jurkat T-cell line. CD147 knockdown resulted in the decreased proliferation and migration of Jurkat cells and reduced the adhesion of Jurkat cells to extracelluar matrix fibronectin in vitro...
August 2008: Cancer Investigation
Majid Momeny, Mohammad R Khorramizadeh, Seyed H Ghaffari, Mehdi Yousefi, Mir S Yekaninejad, Rezvan Esmaeili, Zahra Jahanshiri, Mohammad R Nooridaloii
The purpose of the current study is to evaluate the effect of silibinin on human hepatocellular carcinoma HepG-2 cells. Microculture tetrazolium test (MTT assay), Lactate dehydrogenase (LDH) release, Gelatin zymography, Griess reaction, Cell-based the extracelluar signal-regulated kinase (ERK) 1/2 phosphorylation assay and quantitative real-time RT-PCR were employed to appraise the effect of silibinin on cell proliferation, cytotoxicity, metastatic potential, nitric oxide (NO) production, ERK 1/2 phosphorylation and activation in HepG-2 cells...
September 4, 2008: European Journal of Pharmacology
Izumi Yawata, Hideyuki Takeuchi, Yukiko Doi, Jianfeng Liang, Tetsuya Mizuno, Akio Suzumura
We have shown previously, that the most neurotoxic factor from activated microglia is glutamate that is produced by glutaminase utilizing extracellular glutamine as a substrate. Drugs that inhibit glutaminase or gap junction through which the glutamate is released were effective in reducing neurotoxic activity of microglia. In this study, to elucidate whether or not a similar mechanism is operating in macrophages infiltrating into the central nervous system during inflammatory, demyelinating, and ischemic brain diseases, we examined the neurotoxicity induced by macrophages, in comparison with microglia in vitro...
May 23, 2008: Life Sciences
H Wang, X M Jiang, J H Xu, J Xu, J X Tong, Y W Wang
Mesangioproliferative glomerulonephritis (MsPGN) is a disease of high incidence in humans. Rats with Thy-1 nephritis (Thy-1 N) are used as an animal model for studying MsPGN. Although several studies have demonstrated that many pathological factors could cause the injury of glomerular mesangial cells (GMCs) in Thy-1 N, changes of profile and the molecular mechanism of the disease (i.e. the role of transcription factors) at intervals remain unclear. The purpose of this study was to identify the changes in gene expression profile and to observe the role of nuclear factor kappa B (NF-kappaB) on the pathological change of renal tissue in Thy-1 N rats...
June 2008: Clinical and Experimental Immunology
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