Construction and Validation of a Dual-Transgene Vector System for Stable Transformation in Plants

Publication date: Available online 10 March 2016
Source:Journal of Genetics and Genomics
Author(s): Zhimin He, Bin Liu, Xu Wang, Mingdi Bian, Reqing He, Jindong Yan, Ming Zhong, Xiaoying Zhao, Xuanming Liu
In this study, we constructed dual-transgene vectors (pDT1, pDT7, and pDT7G) that simultaneously co-expressed two genes for use in plants, and ACTIN2 and UBQ10 promoters were used to control the expression of these two genes. The use of 4×Myc, 3×HA, and 3×Flag reporter genes allowed for the convenient identification of a tunable co-expression system in plants, whereas the dexamethasone (Dex) inducible reporter gene c-terminus of the glucocorticoid receptor (cGR) provided Dex-dependent translocation of the fusion gene between the nucleus and cytoplasm. The function of pDT vectors was validated using four pairwise genes in Nicotiana benthamiana or Arabidopsis thaliana. The co-expression efficiency of two genes from the pDT1 and pDT7G vectors was 35% and 42%, respectively, which ensured the generation of sufficient transgenic materials. These pDT vectors are simple, reliable, efficient, and time-saving tools for the co-expression of two genes through a single transformation event and can be used in the study of protein-protein interactions or multi-component complexes.



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Non-invasive Assessment of NASH and NASH-related Fibrosis: is There a Role for Cytokeratin 18?.

No abstract available

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Are We Overdoing Pediatric Lower Gastrointestinal Endoscopy?.

Introduction: Lower gastrointestinal endoscopy (LGIE)/colonoscopy is frequently performed for rectal bleeding, recurrent abdominal pain and the diagnosis of Inflammatory Bowel Disease (IBD). Although these are common indications, the yield for isolated rectal bleeding and recurrent abdominal pain in the otherwise well child has not been described. Methods: A retrospective analysis of patients who had had a LGIE/colonoscopy from January 2001 - December 2010 was performed. The following data were collected: demographic data, indication, distance reached, macroscopic findings, microscopic findings, diagnosis, additional procedures and complications. Results: There were a total of 999 colonoscopies. The colonoscopy was normal in 390/999 (39%). The commonest indication for colonoscopy was for a diagnosis of suspected IBD, 449/999 (45%). IBD was confirmed in 282/449 (63%) but colonoscopy was normal in 143/449 (32%) of suspected IBD. Colonoscopy was performed for rectal bleeding in 197/999 (20%) of whom 141/197 (72%) were normal. There were 46 (5%) colonoscopies performed for recurrent abdominal pain which were all normal. Our completion rate to the cecum and beyond was 521/999 (52%). Our perforation rate over the 10 years was 0.2%. Conclusions: Colonoscopy is a safe procedure in pediatrics, however thirty-nine percent of colonoscopies in this series were normal. Many of these could have been avoided by eliminating colonoscopy in patients with recurrent abdominal pain in the absence of other clinical features, conservative management with laxatives for those with fresh blood per rectum typical of anal fissures, and fecal calprotectin screening prior to endoscopy in patients with suspected IBD. (C) 2016 by European Society for Pediatric Gastroenterology, Hepatology, and Nutrition and North American Society for Pediatric Gastroenterology,

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Identification of Circulating MicroRNAs in Biliary Atresia by Next-Generation Sequencing.

Objectives: Biliary atresia (BA) is an idiopathic neonatal liver disease, characterized by inflammatory and fibrotic obliteration of extrahepatic bile ducts. Therefore, reliable methods for noninvasive diagnosis are needed. This study aimed to analyze circulating microRNAs (miRNAs) in patients with BA using next-generation sequencing (NGS) for identifying novel diagnostic biomarkers. Methods: An initial screening of miRNAs in plasma from patients with BA and healthy controls (HCs) was performed on an Illumina NGS platform. Differential miRNAs were validated by quantitative real-time PCR (qPCR). Target genes and related signal transduction pathways of differential miRNAs were predicted by online software. Results: In total, 146 differential miRNAs were identified by deep sequencing. Fifteen miRNAs with read counts more than 1000, that included 7 upregulated and 8 downregulated miRNAs, were predicted to be associated with liver fibrosis, biliary differentiation and bile duct development. Of these, six miRNAs with read counts more than 5000 were analyzed by qPCR on an independent sample set comprising 44 patients with BA, 20 cholestatic disease controls and 20 HCs. Two up-regulated miRNAs (miR-122-5p, miR-100-5p) and two down-regulated miRNAs (miR-140-3p, miR-126-3p) were confirmed by individual qPCR. Only miR-140-3p was significantly different from controls (P

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Influence of Partial External Biliary Diversion on the Lipid Profile in Children with PFIC.

Background and aim: The concentration of bile acids is highly increased in progressive familial intrahepatic cholestasis (PFIC). Bile acids are the end products of cholesterol metabolism, and aid in the absorption of fat-soluble vitamins and dietary fat. The aim of our study was to investigate lipid metabolism in PFIC patients with focus on the effect of partial external biliary diversion (PEBD). Methods: In 26 patients with PFIC, who underwent PEBD surgery at the median age of 2.2 years (range: 0.4-16.6),we analyzed the concentrations of lipids and apolipoproteins both before and 6 months after PEBD. Patients were split into two groups according to the outcome of surgery (either "good" or "poor"), and were analyzed separately. A "good" result following surgery was defined as complete relief from pruritus, and normalization of total bilirubin ( 190 mg/dl) in 13 patients, phospholipids were increased (>250 mg/dl) in five patients, and triglyceride concentration was increased (>150 mg/dl) in 13 patients. After PEBD, the concentrations of plasma cholesterol, triglycerides, and phospholipids decreased significantly; whereas, ApoA-I and HDL-C concentrations increased and the concentrations of ApoB, LDL-C and VLDL-C significantly decreased. PEBD had neither an effect on ApoE concentration nor on LCAT activity. In the group with a "poor" outcome report following PEBD, total serum cholesterol concentration decreased significantly, and no effect on the concentrations of triglycerides and phospholipids were observed. Conclusion: Patients with PFIC present with a high risk of lipid disturbances. PEBD has a beneficial effect on lipid profile in the majority of cases. (C) 2016 by European Society for Pediatric Gastroenterology, Hepatology, and Nutrition and North American Society for Pediatric Gastroenterology,

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Seattle FD Building Explosion Audio

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The role of octamer binding transcription factors in glioblastoma multiforme

Publication date: Available online 8 March 2016
Source:Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms
Author(s): A.K. Rooj, A. Bronisz, J. Godlewski
A group of transcription factors (TF) that are master developmental regulators of the establishment and maintenance of pluripotency during embryogenesis play additional roles to control tissue homeostasis and regeneration in adults. Among these TFs, members of the Octamer-binding transcription factor (OCT) gene family are well documented as major regulators controlling the self-renewal and pluripotency of stem cells isolated from different adult organs including the brain. In the last few years a large number of studies show the aberrant expression and dysfunction of OCT in different types of cancers including glioblastoma multiforme (GBM). GBM is the most common malignant primary brain tumor, and contains a subpopulation of undifferentiated stem cells (GSCs), with self-renewal and tumorigenic potential that contribute to tumor initiation, invasion, recurrence, and therapeutic resistance. In this review, we have summarized the current knowledge about OCT family in GBM and their crucial role in the initiation, maintenance and drug resistance properties of GSCs. This article is part of a Special Issue entitled: The Oct Transcription Factor Family, edited by Dr. Dean Tantin.



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