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https://www.readbyqxmd.com/read/29244783/using-a-commonly-down-regulated-cytomegalovirus-cmv-promoter-for-high-level-expression-of-ectopic-gene-in-a-human-b-lymphoma-cell-line
#1
Xiaojun Yu, Wei Geng, Hongchuan Zhao, Guobin Wang, Yijun Zhao, Zhe Zhu, Xiaoping Geng
BACKGROUND Vectors are widely used to drive gene expression using a promoter. However, not all promoters are able to drive ectopic gene expression efficiently, including CMV promoter. Here, we report our data using CMV promoter for high-level gene expression in a B lymphoma cell line DG75. MATERIAL AND METHODS A plasmid (pcDNA3.1(+)) containing the CD21 gene driven under CMV promoter was constructed. The plasmid was stably transfected into a human B lymphoma cell line DG75 for cellular surface CD21 expression, and flow cytometry was used to monitor CD21 expression...
December 15, 2017: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
https://www.readbyqxmd.com/read/29244483/biomembrane-permeabilization-statistics-of-individual-leakage-events-harmonize-the-interpretation-of-vesicle-leakage
#2
Stefan Braun, Šárka Pokorná, Radek Šachl, Martin Hof, Heiko Heerklotz, Maria Hoernke
The mode of action of membrane-active molecules such as antimicrobial, anticancer, cell penetrating, and fusion peptides and their synthetic mimics, transfection agents and drug permeation enhancers, biological signaling molecules (e.g., quorum sensing), involves either the general or local destabilization of the target membrane or the formation of defined, rather stable pores. Some effects aim at killing the cell, others need to be limited in space and time to avoid serious damage. Biological tests reveal translocation of compounds and cell death but do not provide a detailed, mechanistic, and quantitative understanding of the modes of action and their molecular basis...
December 15, 2017: ACS Nano
https://www.readbyqxmd.com/read/29244234/mechanisms-of-caspr2-antibodies-in-autoimmune-encephalitis-and-neuromyotonia
#3
Kristina R Patterson, Josep Dalmau, Eric Lancaster
OBJECTIVE: To determine the pathogenic mechanisms of autoantibodies to the cell adhesion molecule Caspr2 in acquired neuromyotonia and autoimmune encephalitis. METHODS: Caspr2 positive samples were confirmed using a cell-based assay, and their IgG subtypes were determined by ELISA and cell-based assay. A solid phase binding assay quantified the binding of Caspr2 to contactin-2 in the presence of Caspr2 autoantibodies. Living cultures of primary rat hippocampal neurons were incubated with Caspr2-positive or control sera and the distribution of Caspr2-positive immunofluorescent puncta and total surface Caspr2 were quantified...
December 15, 2017: Annals of Neurology
https://www.readbyqxmd.com/read/29244209/generation-of-a-kshv-k13-deletion-mutant-for-vflip-function-study
#4
Fei Wang, Yuanyuan Guo, Wan Li, Chun Lu, Qin Yan
Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded viral Fas-associated death domain-like IL-1-converting enzyme inhibitory protein (vFLIP) is one of the latently expressed genes and plays a key role in cell survival and maintenance of latent infection by activating the NF-κB pathway. To obtain a genetic system for studying KSHV vFLIP mutation in the context of the viral genome, we generated recombinant viruses lacking the coding sequence (CDS) of vFLIP gene (K13/ORF71) by bacterial artificial chromosome (BAC) technology and the Escherichia coli Red recombination system...
December 15, 2017: Journal of Medical Virology
https://www.readbyqxmd.com/read/29243846/aqp2-can-modulate-the-pattern-of-ca2-transients-induced-by-store-operated-ca2-entry-under-trpv4-activation
#5
Alejandro Pizzoni, Macarena López González, Gisela Di Giusto, Valeria Rivarola, Claudia Capurro, Paula Ford
There is increasing evidence indicating that aquaporins (AQPs) exert an influence in cell signaling by the interplay with the TRPV4 Ca2+ channel. Ca2+ release from intracellular stores and plasma membrane hyperpolarization due to opening of Ca2+ -activated potassium channels (KCa) are events that have been proposed to take place downstream of TRPV4 activation. A major mechanism for Ca2+ entry, activated after depletion of intracellular Ca2+ stores and driven by electrochemical forces, is the store-operated Ca2+ entry (SOCE)...
December 15, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/29243812/dysregulated-mitogen-activated-protein-kinase-signalling-as-an-oncogenic-basis-for-clear-cell-sarcoma-of-the-kidney
#6
Colin Kenny, Naomi McDonagh, Antonio Lazaro, Elaine O'Meara, Rut Klinger, Darran O'Connor, Fiona Roche, Karsten Hokamp, Maureen J O'Sullivan
The oncogenic mechanisms and tumour biology underpinning Clear Cell Sarcoma of Kidney (CCSK), the second commonest paediatric renal malignancy, are poorly understood and currently therapy depends heavily on doxorubicin with cardiotoxic side-effects. Previously, we characterised the balanced t(10;17)(q22;p13) chromosomal translocation, identified at that time as the only recurrent genetic aberration in CCSK. This translocation results in an in-frame fusion of the genes YWHAE (encoding 14-3-3ε)and NUTM2, with a somatic incidence of 12%...
December 15, 2017: Journal of Pathology
https://www.readbyqxmd.com/read/29243790/mir-93-inhibition-ameliorates-ogd-r-induced-cardiomyocyte-apoptosis-by-targeting-nrf2
#7
L-J Yan, X-W Fan, H-T Yang, J-T Wu, S-L Wang, C-G Qiu
OBJECTIVE: To identify the expression changes of microRNA 93 (miR-93) in oxygen-glucose deprivation/reoxygenation (OGD/R) injury in cardiomyocytes and its mechanism of mediating OGD/R and inducing apoptosis. MATERIALS AND METHODS: Primary cardiomyocytes were extracted and OGD/R model in cardiomyocytes was established in vitro. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to detect the expressions of miR-93, and Western blot assay was applied to measure the protein levels of nuclear factor erythroid 2-related factor 2 (Nrf2) and caspase-3...
December 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/29243779/effects-of-ngx6-expression-on-proliferation-and-invasion-of-nasopharyngeal-carcinoma-cells-and-survival-of-patients
#8
Y-T Wang, X-Y Sun, H Wang, T-T Huang, Y Wang, M-X Yang
OBJECTIVE: To detect the expressions of nasopharyngeal carcinoma-associated gene 6 (NGX6) in nasopharyngeal carcinoma cells and tissues, and to investigate the effects of NGX6 on the proliferation and invasion of nasopharyngeal carcinoma cells and the survival of patients. PATIENTS AND METHODS: The human nasopharyngeal carcinoma cells (HONE1) and immortalized human nasopharyngeal epithelial cells (NP69) were selected and cultured. The mRNA and protein expression levels of NGX6 were detected via quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot...
December 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/29242628/mutant-runx2-regulates-amelogenesis-and-osteogenesis-through-a-mir-185-5p-dlx2-axis
#9
Huaiguang Chang, Yue Wang, Haochen Liu, Xu Nan, Singwai Wong, Saihui Peng, Yajuan Gu, Hongshan Zhao, Hailan Feng
Regulation of microRNAs (miRNA) has been extensively investigated in diseases; however, little is known about the roles of miRNAs in cleidocranial dysplasia (CCD). The aim of the present study was to investigate the potential involvement of miRNAs in CCD. In vitro site-directed mutagenesis was performed to construct three mutant Runx2 expression vectors, which were then transfected into LS8 cells and MC3T3-E1 cells, to determine the impact on amelogenesis and osteogenesis, respectively. miRCURY LNA miRNA microarray identify miR-185-5p as a miRNA target commonly induced by all three Runx2 mutants...
December 14, 2017: Cell Death & Disease
https://www.readbyqxmd.com/read/29242172/curcumin-enhances-lxr%C3%AE-in-an-amp-activated-protein-kinase-dependent-manner-in-human-macrophages
#10
Javier Sáenz, Gonzalo Alba, María E Reyes-Quiroz, Isabel Geniz, Juan Jiménez, Francisco Sobrino, Consuelo Santa-María
Liver X receptor alpha (LXRα) is a nuclear receptor involved in cholesterol homeostasis. Curcumin, a traditional Chinese derivative from the rhizomes of Curcuma longa and a well-known AMP-activated protein kinase (AMPK) activator, possess hypocholesterolemic activity, however, the possible link between AMPK and cholesterol is unknown. In this study, we have investigated whether curcumin regulates metabolic changes in cholesterol metabolism via LXRα in THP-1 human macrophages, the cells implicated in atheroma plaques formation...
November 16, 2017: Journal of Nutritional Biochemistry
https://www.readbyqxmd.com/read/29242061/phosphorylation-of-ser-525-in-%C3%AE-pix-impairs-nox1-activating-ability-in-caco-2%C3%A2-cells
#11
Yuuki Kaito, Ryosuke Kataoka, Tatsuya Mihara, Kento Takechi, Akira Takahira, Shuhei Watanabe, Fei Han, Minoru Tamura
βPix activates Nox1, an O2--generating NADPH oxidase, through Rac activation. In this study, we found that S525E mutation of βPix eliminated its Nox1-activating ability in transfected Caco-2 cells. Unexpectedly, affinity for Rac was not diminished but rather enhanced by S525E mutation, and guanine nucleotide exchange factor (GEF) activity was not altered. The N-terminal fragment (amino acids 1-400) showed similar Rac-binding and GEF activity to wild-type βPix. In contrast, the C-terminal fragment (amino acids 408-646) had higher Rac-binding activity, particularly for Rac-GTP, than wild-type βPix, and showed no GEF activity...
December 11, 2017: Archives of Biochemistry and Biophysics
https://www.readbyqxmd.com/read/29241219/yap-inhibits-the-apoptosis-and-migration-of-human-rectal-cancer-cells-via-suppression-of-jnk-drp1-mitochondrial-fission-htra2-omi-pathways
#12
Haijun Li, Fucheng He, Xin Zhao, Yuan Zhang, Xi Chu, Chunlan Hua, Yunhui Qu, Yu Duan, Liang Ming
BACKGROUND/AIMS: The Hippo-Yap pathway is associated with tumor development and progression. However, little evidence is available concerning its role in cancer cell apoptosis and migration via mitochondrial homeostasis. Here, we identify mitochondrial fission as a regulator of the Hippo-Yap pathway in human rectal cancer tumorigenesis and metastasis. METHODS: In this study, we performed loss-of function assays concerning Yap in RCC via shRNA. Cellular viability and apoptosis were measured via MTT, the TUNEL assay and trypan blue staining...
December 12, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29241198/fam3a-protects-against-glutamate-induced-toxicity-by-preserving-calcium-homeostasis-in-differentiated-pc12-cells
#13
Qing Song, Wen-Li Gou, Yu-Liang Zou
BACKGROUND/AIMS: Stroke is the leading cause of adult disability, and glutamate-induced dysregulation of intracellular Ca2+ homeostasis is a key mechanism. FAM3A is the first member of the family with sequence similarity 3 (FAM3) gene family, and its biological function remains largely unknown. We have recently reported that FAM3A exerts protective effects against oxidative stress and mitochondrial dysfunction in HT22 cells. METHODS: Here, we investigated the protective effects of FAM3A using a glutamate-induced neuronal injury model in nerve growth factor (NGF)-differentiated PC12 cells...
December 12, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29241192/microrna-92a-3p-regulates-aggrecanase-1-and-aggrecanase-2-expression-in-chondrogenesis-and-il-1%C3%AE-induced-catabolism-in-human-articular-chondrocytes
#14
Guping Mao, Peihui Wu, Ziji Zhang, Zhiqi Zhang, Weiming Liao, Yukang Li, Yan Kang
BACKGROUND/AIMS: Aggrecanase-1 (ADAMTS-4) and aggrecanase-2 (ADAMTS-5) are secreted enzymes belonging to the ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs) family that play significant roles in the progression of osteoarthritis (OA). Here, we aimed to determine whether the expression of ADAMTS-4/5 in chondrogenesis and inflammation is regulated by microRNA-92a-3p (miR-92a-3p). METHODS: MiR-92a-3p and ADAMTS-4/5 expressions were determined by quantitative polymerase chain reaction (qPCR)...
November 3, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29241179/rs739837-polymorphism-in-mir-885-3p-binding-site-within-3-untranslated-region-of-vitamin-d-receptor-is-associated-with-a-decreased-risk-of-pressure-ulcers
#15
Xia Ji, Jinbao Mao, Shuang Zhou
BACKGROUND/AIMS: Accumulative evidence has shown that miR-885-3p plays a crucial role in human carcinogenesis. From miRNA database, we also found that rs739837 polymorphism in miR-885-3p binding site within 3'-untranslated region of Vitamin D receptor (VDR), compromising the suppressive effect of miR-885-3p on VDR. Moreover, Vitamin D is involved in controlling the cell immune response and may play a role in pressure ulcers development. However, whether this polymorphism is actually linked with pressure ulcers remains unclear...
December 12, 2017: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29241074/sall4-promotes-osteoblast-differentiation-by-deactivating-notch2-signaling
#16
Wei Yu, Dianming Jiang, Yu Shaobin, Fu Jiabin, Zhijun Li, Yuchi Wu, Yulong Wang
BACKGROUND: The Spalt-Like Transcription Factor 4 (SALL4) is reported to regulate cell proliferation, migration and invasion. However, the roles of SALL4 in osteoblast differentiation are unclear. This study was aimed to explore the underlying mechanism of SALL4 in osteoporosis. METHODS: Firstly, the expression of SALL4 was assessed in vivo and in vitro at various stages of development of rats (E14, E20, postnatal 2, 4, and 9 day) or different incubation time (0, 6, 9, 12 and 15 day) of C2C12 and MC3T3-E1 cells...
December 11, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
https://www.readbyqxmd.com/read/29241031/the-protective-effect-of-dexmedetomidine-on-lps-induced-acute-lung-injury-through-the-hmgb1-mediated-tlr4-nf-%C3%AE%C2%BAb-and-pi3k-akt-mtor-pathways
#17
Lu Meng, Longyun Li, Shan Lu, Kai Li, Zhenbo Su, Yunyun Wang, Xiaodi Fan, Xuyang Li, Guoqing Zhao
The aim of present study was to evaluate the protective effects of dexmedetomidine (DEX) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) and investigate its possible mechanisms mediated by HMGB1. In vivo, pulmonary pathology observation and myeloperoxidase (MPO) activity were also examined to evaluate the protective effect of DEX in the lungs. Tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) in bronchoalveolar lavage fluid (BALF), serum and lung tissues LPS-induced rats were detected...
December 11, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/29240831/comparative-transcriptome-profiling-of-virulent-and-non-virulent-trypanosoma-cruzi-underlines-the-role-of-surface-proteins-during-infection
#18
A Trey Belew, Caroline Junqueira, Gabriela F Rodrigues-Luiz, Bruna M Valente, Antonio Edson R Oliveira, Rafael B Polidoro, Luciana W Zuccherato, Daniella C Bartholomeu, Sergio Schenkman, Ricardo T Gazzinelli, Barbara A Burleigh, Najib M El-Sayed, Santuza M R Teixeira
Trypanosoma cruzi, the protozoan that causes Chagas disease, has a complex life cycle involving several morphologically and biochemically distinct stages that establish intricate interactions with various insect and mammalian hosts. It has also a heterogeneous population structure comprising strains with distinct properties such as virulence, sensitivity to drugs, antigenic profile and tissue tropism. We present a comparative transcriptome analysis of two cloned T. cruzi strains that display contrasting virulence phenotypes in animal models of infection: CL Brener is a virulent clone and CL-14 is a clone that is neither infective nor pathogenic in in vivo models of infection...
December 14, 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/29240260/sohlh1-suppresses-glioblastoma-cell-proliferation-migration-and-invasion-by-inhibition-of-wnt-%C3%AE-catenin-signaling
#19
Xinyu Liu, Qing Gao, Na Zhao, Xiaoli Zhang, Weiwei Cui, Jinhao Sun, Jiang Fu, Jing Hao
Glioblastoma (GBM) is the most aggressive and highly invasive type of astrocytic tumors. Despite advances in diagnosis and therapy, the prognosis and survival time remain poor. Identifying key mediators of tumor cell proliferation, migration and invasion is crucial to the development of new and more effective therapies. In this paper, we report the novel role of Spermatogenesis- and oogenesis- specific basic helix-loop-helix transcription factor1 (Sohlh1) in the inhibition of Wnt/β-catenin signaling and aggressive behaviors in GBM cells...
December 14, 2017: Molecular Carcinogenesis
https://www.readbyqxmd.com/read/29239816/mir-9-5p-promotes-cell-growth-and-metastasis-in-non-small-cell-lung-cancer-through-the-repression-of-tgfbr2
#20
Gang Li, Fang Wu, Han Yang, Xia Deng, Yawei Yuan
BACKGROUND: Increasing evidence indicates that the dysregulation of microRNAs (miRNAs) play critical roles tumor progression and metastasis, but very few papers had reported the function of miR-9-5p in lung cancer, especially in NSCLCs. METHODS: In this study, we investigated the role of miR-9-5p in non-small cell lung cancers (NSCLCs). MiR-9-5p level were analyzed in 62 clinical NSCLC lung tissue samples and adjacent normal lung tissues by RT-PCR. The target of miR-9-5p was predicted by TargetScan and luciferase reporter assay was used to verify the binding site of miR-9-5p on TGFBR2 mRNA...
November 24, 2017: Biomedicine & Pharmacotherapy, Biomédecine & Pharmacothérapie
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