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https://www.readbyqxmd.com/read/29344663/adeno-associated-virus-type-2-mediated-gene-transfer-of-a-short-hairpin-rna-targeting-human-igfbp-2-suppresses-the-proliferation-and-invasion-of-mda-mb-468-cells
#1
Chao Gao, Ru-Song Zhang, Nan Zheng, Chen Wang
Adeno-associated virus 2 (AAV2) is prepotent in the biological treatment of breast tumor because of its low pathogenicity and immunogenicity. Our previous study demonstrated that insulin‑like growth factor‑binding protein 2 (IGFBP‑2) was highly expressed in patients with breast metastasis. In the present study, the effects of recombinant AAV2 on the growth and metastasis of breast cancer cells were determined in vitro, and in vivo. rAAV2-ZsGreen-shRNA-scramble and rAAV2‑ZsGreen‑shRNA‑hIGFBP‑2 were used to transfect MDA‑MB‑468, and MCF‑10A cells respectively, and observed that these virus could not penetrate the normal human breast epithelia MCF‑10A cell line...
January 16, 2018: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29276753/indian-hedgehog-suppresses-a-stromal-cell-driven-intestinal-immune-response
#2
B Florien Westendorp, Nikè V J A Büller, Olga N Karpus, Willemijn A van Dop, Jan Koster, Rogier Versteeg, Pim J Koelink, Clinton Y Snel, Sander Meisner, Joris J T H Roelofs, Anja Uhmann, Emiel Ver Loren van Themaat, Jarom Heijmans, Heidi Hahn, Vanesa Muncan, Manon E Wildenberg, Gijs R van den Brink
Background & Aims: Upon intestinal epithelial damage a complex wound healing response is initiated to restore epithelial integrity and defend against pathogenic invasion. Epithelium-derived Indian Hedgehog (Ihh) functions as a critical sensor in this process. Signaling occurs in a paracrine manner because the receptor for Ihh is expressed only in the mesenchyme, but the exact Hedgehog target cell has remained elusive. The aim of this study was to elucidate further the nature of this target cell in the context of intestinal inflammation...
2018: Cellular and Molecular Gastroenterology and Hepatology
https://www.readbyqxmd.com/read/29232382/charting-the-transcriptional-landscape-of-cells-of-renin-lineage-following-podocyte-depletion
#3
Aaron D McClelland, Julia Lichtnekert, Diana G Eng, Jeffrey W Pippin, Kenneth W Gross, Sina A Gharib, Stuart J Shankland
Renin producing cells of the juxtaglomerulus, herein called cells of renin lineage (CoRL), have garnered recent interest for their propensity to act as a progenitor source for various kidney cell types including podocytes. Despite recent advances, the process of transdifferentiation of CoRL to podocytes is poorly understood. In this study, we employed a transgenic reporter mouse line which permanently labels CoRL with ZsGreen fluorescent protein, allowing for isolation by fluorescence-activated cell sorting...
2017: PloS One
https://www.readbyqxmd.com/read/29152658/hepatic-differentiation-of-mouse-bone-marrow%C3%A2-derived-mesenchymal-stem-cells-using-a-novel-3d-culture-system
#4
Qiong Wu, Jing Tang, Yi Li, Li Li, Yujia Wang, Ji Bao, Hong Bu
The development of novel culture systems that mimic the in vivo microenvironment may be beneficial for inducing the differentiation of stem cells and promoting liver function. In the present study, spheroid cultures and decellularized liver scaffolds (DLSs) were utilized to obtain differentiated hepatocyte‑like cells. Mouse bone marrow (BM)‑derived mesenchymal stem cells (MSCs) self‑aggregated into spheroids under low‑attachment conditions and implanted into the DLSs via a negative pressure suction device...
October 19, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/29131531/modifying-expression-modes-of-human-neurotensin-receptor-type-1-alters-sensing-capabilities-for-agonists-in-yeast-signaling-biosensor
#5
Hiroki Hashi, Yasuyuki Nakamura, Jun Ishii, Akihiko Kondo
Neurotensin receptor type 1 (NTSR1), a member of the G-protein-coupled receptor (GPCR) family, is naturally activated by binding of a neurotensin peptide, leading to a variety of physiological effects. The budding yeast Saccharomyces cerevisiae is a proven host organism for assaying the agonistic activation of human GPCRs. Previous studies showed that yeast cells can functionally express human NTSR1 receptor, permitting the detection of neurotensin-promoted signaling using a ZsGreen fluorescent reporter gene...
November 13, 2017: Biotechnology Journal
https://www.readbyqxmd.com/read/29097083/knockdown-of-melatonin-receptor-1-and-induction-of-follicle-stimulating-hormone-on-the-regulation-of-mouse-granulosa-cell-function
#6
Hira Sajjad Talpur, Tesfaye Worku, Zia Ur Rehman, Rahim Dad, Dinesh Bhattarai, Iqra Bano, Farmanullah, Aixin Liang, Changjiu He, Liguo Yang
Melatonin receptor 1 (MT1) performs a critical role in the regulation of the animal reproductive system, particularly in follicular growth, and has a considerable effect on reproductive performance. However, the role that MT1 plays in regulating hormones associated with reproduction remains unclear. This study was designed to examine the physiological role of constitutive MT1 silencing and follicle stimulating hormone (FSH) treatment in reproduction, making use of mouse granulosa cells (mGCs) as a model. To understand the constitutive role of MT1 in ovarian physiology, the RNAi-Ready pSIREN-RETROQ-ZsGreen Vector mediated recombinant pshRNA was used to silence MT1 gene expression...
October 30, 2017: Reproductive Biology
https://www.readbyqxmd.com/read/29016733/endothelial-to-haematopoietic-transition-contributes-to-pulmonary-arterial-hypertension
#7
Olin D Liang, Eui-Young So, Pamela C Egan, Laura R Goldberg, Jason M Aliotta, Keith Q Wu, Patrycja M Dubielecka, Corey E Ventetuolo, Anthony M Reginato, Peter J Quesenberry, James R Klinger
Aims: The pathogenic mechanisms of pulmonary arterial hypertension (PAH) remain unclear, but involve dysfunctional endothelial cells (ECs), dysregulated immunity and inflammation in the lung. We hypothesize that a developmental process called endothelial to haematopoietic transition (EHT) contributes to the pathogenesis of pulmonary hypertension (PH). We sought to determine the role of EHT in mouse models of PH, to characterize specific cell types involved in this process, and to identify potential therapeutic targets to prevent disease progression...
November 1, 2017: Cardiovascular Research
https://www.readbyqxmd.com/read/28879651/preliminary-studies-on-the-differential-expression-of-adrenocorticotropic-hormone-receptor-in-adrenal-adenomas
#8
Dongliang Hu, Tongzu Liu, Xinghuan Wang
INTRODUCTION: The ACTH receptor (ACTHR) is primarily expressed in the adrenal cortex. Previous studies focused on the regulatory function of ACTHR in glucocorticoid secretion, but research on adrenal tumours is rare. The aim of this study was to evaluate ACTHR expression in common adrenal adenomas and investigate its influence on adrenal tumorigenesis using adrenocortical H295R cells. MATERIALS AND M: ETHODS: Real-time polymerase chain reaction and western blot were used to detect the expression of ACTHR in 18 aldosterone-producing adenomas, 16 cortisol-producing adenomas, nine non-functional adenomas, and 12 normal adrenal samples...
September 7, 2017: Endokrynologia Polska
https://www.readbyqxmd.com/read/28745324/a-committed-postselection-precursor-to-natural-tcr%C3%AE-%C3%AE-intraepithelial-lymphocytes
#9
Christoph S N Klose, Jonas F Hummel, Lena Faller, Yannick d'Hargues, Karolina Ebert, Yakup Tanriver
The intestine is a major immune organ with several specialized lymphoid structures and immune cells. Among these are thymus-derived natural intraepithelial lymphocytes (IELs) that lack expression of the classical co-receptors CD4 or CD8αβ (double negative (DN)). Natural IELs are both αβ(+) and γδ(+) T cells that play important roles in the maintenance of the epithelial barrier at steady state and during inflammation. The transcription factor T-bet is essential for the peripheral development of natural IELs, but its role during thymic development has remained less clear...
July 26, 2017: Mucosal Immunology
https://www.readbyqxmd.com/read/28681159/selectable-marker-independent-transformation-of-recalcitrant-maize-inbred-b73-and-sorghum-p898012-mediated-by-morphogenic-regulators-baby-boom-and-wuschel2
#10
Muruganantham Mookkan, Kimberly Nelson-Vasilchik, Joel Hague, Zhanyuan J Zhang, Albert P Kausch
Discriminatory co-expression of maize BBM and WUS transcriptional factor genes promoted somatic embryogenesis and efficient Agrobacterium -mediated transformation of recalcitrant maize inbred B73 and sorghum P898012 genotypes without use of a selectable marker gene. The use of morphogenic regulators to overcome barriers in plant transformation is a revolutionary breakthrough for basic plant science and crop applications. Current standard plant transformation systems are bottlenecks for genetic, genomic, and crop improvement studies...
July 5, 2017: Plant Cell Reports
https://www.readbyqxmd.com/read/28617813/identification-of-padi2-as-a-novel-angiogenesis-regulating-gene-by-genome-association-studies-in-mice
#11
Mehrdad Khajavi, Yi Zhou, Amy E Birsner, Lauren Bazinet, Amanda Rosa Di Sant, Alex J Schiffer, Michael S Rogers, Subrahmanian Tarakkad Krishnaji, Bella Hu, Vy Nguyen, Leonard Zon, Robert J D'Amato
Recent findings indicate that growth factor-driven angiogenesis is markedly influenced by genetic variation. This variation in angiogenic responsiveness may alter the susceptibility to a number of angiogenesis-dependent diseases. Here, we utilized the genetic diversity available in common inbred mouse strains to identify the loci and candidate genes responsible for differences in angiogenic response. The corneal micropocket neovascularization assay was performed on 42 different inbred mouse strains using basic fibroblast growth factor (bFGF) pellets...
June 2017: PLoS Genetics
https://www.readbyqxmd.com/read/28597477/effective-expression-of-drebrin-in-hippocampus-improves-cognitive-function-and-alleviates-lesions-of-alzheimer-s-disease-in-app-swe-ps1-%C3%AE-e9-mice
#12
Yan Liu, Yan-Feng Xu, Ling Zhang, Lan Huang, Pin Yu, Hua Zhu, Wei Deng, Chuan Qin
AIMS: Alzheimer's disease (AD), a progressive development dementia, is increasingly impacting patients' living conditions worldwide. Despite medical care and funding support, there are still no highly individualized drugs and practical strategies for clinical prevention and treatment. Developmentally regulated brain protein (abbreviated as Drebrin or Dbn, also known as Dbn1 in mouse) exists in neurons, especially in dendrites, and is an actin-binding protein that modulates synaptic morphology and long-term memory...
July 2017: CNS Neuroscience & Therapeutics
https://www.readbyqxmd.com/read/28584633/construction-of-a-general-albumin-promoter-reporter-system-for-real-time-monitoring-of-the-differentiation-status-of-functional-hepatocytes-from-stem-cells-in-mouse-rat-and-human
#13
Jing Tang, Qiong Wu, Yi Li, Xiujuan Wu, Yujia Wang, Lihua Zhu, Yujun Shi, Hong Bu, Ji Bao, Mingjun Xie
Genetic constructs with promoters fused to reporter genes for simultaneous monitoring of cellular events have been the focus of attention in recent years. Adenoviral vectors, which have distinctive characteristics, have been used to monitor the differentiation of stem cells in vitro. In the present study, a modified adenoviral vector was constructed, containing a mouse, rat, and human general albumin promoter sequence fused to a ZsGreen reporter gene, and evaluated its efficiency in different cell types. Two hepatocyte cell lines (Hepa1-6 and HepG2), rat primary hepatocytes, rat bone marrow mesenchymal stem cells (BM-MSCs) and rat BM-MSCs-derived hepatocyte-like cells were transduced with this vector, and the transfection efficiency and functional capabilities of the promoter were evaluated by fluorescent microscopy...
June 2017: Biomedical Reports
https://www.readbyqxmd.com/read/28583151/identification-and-validation-of-a-schistosoma-japonicum-u6-promoter
#14
Qing Li, Wan Wang, Nan Zhao, Pengcheng Li, Yue Xin, Wei Hu
BACKGROUND: RNA polymerase III promoters have been widely used to express short hairpin-RNA (shRNA), microRNA (miRNA), and small guide RNA (sgRNA) in gene functional analysis in a variety of organisms including Schistosoma mansoni. However, no endogenous RNA polymerase III promoters have been identified in Schistosoma japonicum. The lack of appropriate promoters in S. japonicum has hindered its gene functional analysis. Identification of functional promoters in S. japonicum is therefore in urgent need...
June 5, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28535287/a-flow-cytometry-based-screen-identifies-mbnl1-modulators-that-rescue-splicing-defects-in-myotonic-dystrophy-type-i
#15
Fan Zhang, Nicole E Bodycombe, Keith M Haskell, Yumei L Sun, Eric T Wang, Carl A Morris, Lyn H Jones, Lauren D Wood, Mathew T Pletcher
Myotonic dystrophy Type 1 (DM1) is a rare genetic disease caused by the expansion of CTG trinucleotide repeats ((CTG)exp) in the 3' untranslated region of the DMPK gene. The repeat transcripts sequester the RNA binding protein Muscleblind-like protein 1 (MBNL1) and hamper its normal function in pre-mRNA splicing. Overexpressing exogenous MBNL1 in the DM1 mouse model has been shown to rescue the splicing defects and reverse myotonia. Although a viable therapeutic strategy, pharmacological modulators of MBNL1 expression have not been identified...
August 15, 2017: Human Molecular Genetics
https://www.readbyqxmd.com/read/28339054/overexpression-of-tim-3-reduces-helicobacter-pylori-associated-inflammation-through-tlr4-nf%C3%AE%C2%BAb-signaling-in%C3%A2-vitro
#16
Fucai Wang, Zhirong Mao, Dongsheng Liu, Jing Yu, Youhua Wang, Wen Ye, Dongjia Lin, Nanjin Zhou, Yong Xie
The present study aimed to investigate the interaction between T-cell immunoglobulin and mucin-domain-containing molecule-3 (Tim-3) and Toll-like receptor 4 (TLR4)/nuclear factor κB (NF‑κB) signaling in Helicobacter pylori-infected RAW264.7 macrophage cells. RAW264.7 cells were co‑cultured with H. pylori SS1 at different bacteria/cell ratios, and subsequently the mRNA expression of Tim‑3, TLR4, and myeloid differentiation factor 88 (MyD88) was measured by reverse transcription-quantitative polymerase chain reaction (RT‑qPCR)...
May 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28251288/rnai-as-a-tool-to-control-the-sex-ratio-of-mouse-offspring-by-interrupting-zfx-zfy-genes-in-the-testis
#17
YongSheng Zhang, JiFeng Xi, Bin Jia, XiangZu Wang, XuHai Wang, ChaoCheng Li, YaQiang Li, XianCun Zeng, RuiWen Ying, Xin Li, Song Jiang, FangYuan Yuan
The objective of this study was to explore a novel method to alter the sex-ratio balance of mouse offspring by silencing the paralogous genes Zfx/Zfy (Zinc finger X/Y-chromosomal transcription factor gene) during spermatogenesis. Four recombined vectors PRZ1, PRZ2, PRZ3, and PRZ4 (RNAi-Ready-pSIREN-RetroQ-ZsGreen) were constructed for interrupting the Zfx gene. Additionally, a recombined vector Psilencer/Zfy-shRNA was constructed for interrupting the Zfy gene. Male mice were randomly divided into 8 groups, with 20 animals per group...
April 2017: Mammalian Genome: Official Journal of the International Mammalian Genome Society
https://www.readbyqxmd.com/read/27906115/established-cell-surface-markers-efficiently-isolate-highly-overlapping-populations-of-skeletal-muscle-satellite-cells-by-fluorescence-activated-cell-sorting
#18
Claire C Maesner, Albert E Almada, Amy J Wagers
BACKGROUND: Fluorescent-activated cell sorting (FACS) has enabled the direct isolation of highly enriched skeletal muscle stem cell, or satellite cell, populations from postnatal tissue. Several distinct surface marker panels containing different positively selecting surface antigens have been used to distinguish muscle satellite cells from other non-myogenic cell types. Because functional and transcriptional heterogeneity is known to exist within the satellite cell population, a direct comparison of results obtained in different laboratories has been complicated by a lack of clarity as to whether commonly utilized surface marker combinations select for distinct or overlapping subsets of the satellite cell pool...
November 8, 2016: Skeletal Muscle
https://www.readbyqxmd.com/read/27826411/established-cell-surface-markers-efficiently-isolate-highly-overlapping-populations-of-skeletal-muscle-satellite-cells-by-fluorescence-activated-cell-sorting
#19
Claire C Maesner, Albert E Almada, Amy J Wagers
BACKGROUND: Fluorescent-activated cell sorting (FACS) has enabled the direct isolation of highly enriched skeletal muscle stem cell, or satellite cell, populations from postnatal tissue. Several distinct surface marker panels containing different positively selecting surface antigens have been used to distinguish muscle satellite cells from other non-myogenic cell types. Because functional and transcriptional heterogeneity is known to exist within the satellite cell population, a direct comparison of results obtained in different laboratories has been complicated by a lack of clarity as to whether commonly utilized surface marker combinations select for distinct or overlapping subsets of the satellite cell pool...
2016: Skeletal Muscle
https://www.readbyqxmd.com/read/27771389/lassa-and-ebola-virus-inhibitors-identified-using-minigenome-and-recombinant-virus-reporter-systems
#20
Stephen R Welch, Lisa Wiggleton Guerrero, Ayan K Chakrabarti, Laura K McMullan, Mike Flint, Gregory R Bluemling, George R Painter, Stuart T Nichol, Christina F Spiropoulou, César G Albariño
Lassa virus (LASV) and Ebola virus (EBOV) infections are important global health issues resulting in significant morbidity and mortality. While several promising drug and vaccine trials for EBOV are ongoing, options for LASV infection are currently limited to ribavirin treatment. A major factor impeding the development of antiviral compounds to treat these infections is the need to manipulate the virus under BSL-4 containment, limiting research to a few institutes worldwide. Here we describe the development of a novel LASV minigenome assay based on the ambisense LASV S segment genome, with authentic terminal untranslated regions flanking a ZsGreen (ZsG) fluorescent reporter protein and a Gaussia princeps luciferase (gLuc) reporter gene...
December 2016: Antiviral Research
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