Objective: To clone and express a truncated, soluble vascular endothelial growth factor receptor 2 (sVEGFR2) possessing the combined-functional domains 1-3 and 5 in eukaryotic cells and to test the inhibitory effects of full length VEGFR2 in vivo.
Results: pCMV6-trunctated-rVegfr2 (6100 bp) was successfully cloned. The transfection experiments showed that either pCMV6-truncated-rat-Vegfr2 (pCMV6-truncated-rVegfr2) or pCMV6-rVegfr2 inhibited the expression of intracellular green fluorescent protein, which is usually used as an exogenous transfected reporter gene to determine the transfected efficiency. An analysis of the transfected cells revealed that the amount of full-length VEGFR2 protein in the pCMV6-truncated-rVegfr2 transfected cells was 20% lower than that in the negative control (non-transfected HEK 293 cells). The differences in test results between the transfected and negative control groups were greatest from 24-30 h after transfection; this period was therefore chosen as optimal for collecting culture supernatants. This analysis was highly sensitive for detecting the amount of sVEGFR2 protein expressed and secreted by the cells, and the sVEGFR2 protein content was found to increase by approximately 26% in the transfected cells compared to that in the negative control cells (68.2% ± 1.7% vs. 41.9% ± 2.9%, P = 0.000) and by 18% compared to the negative control cells (68.2% ± 1.7% vs. 50.0% ± 0.5%, P = 0.003). Propidium iodide and Hoechst staining indicated no significant change in the number of HEK293 cells undergoing apoptosis 6 days after pCMV6-trucated-Vegfr2 transfection, compared to the negative control. Soluble VEGFR2 produced by pCMV6-truncated-rVegfr2 inhibited full-length VEGFR2 protein expression in the cell membrane.
Conclusions: This study employed a eukaryotic system to express sVEGFR2. The use of transient transfection technology greatly improved transfect efficiency. Recombinant sVEGFR2 inhibited the effect of endogenous full-length VEGFR2 but was not cytotoxic.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3977943 | PMC |
| http://dx.doi.org/10.1186/2045-3701-4-14 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Comparison of fluoride level between mother's milk and infant formula milk - a global systematic review and dental fluorosis risk assessment.
Food Addit Contam Part A Chem Anal Control Expo Risk Assess
January 2025
Department of Pediatrics and Neonatology, Guangzhou University of Traditional Chinese Medicine Dongguan Hospital, Guangdong Province, China.
Dental Fluorosis (DF) is one of the negative outcomes of excessive fluoride (F) intake through food sources. This systematic review aimed to compare F content in two important food sources for infants, Mother's Milk (MoM) and Infant Formula (IF), and then evaluate the risk of DF related to F in those two types of food. For this purpose, 181 studies were initially found by searching the relevant keywords in widely recognized databases, including Google Scholar, Scopus, Science Direct, and PubMed.
View Article and Find Full Text PDFStress, perceived competence and guilt as predictors of depression in parents with chronic pain.
Front Psychol
January 2025
Department of Psychology, Rey Juan Carlos University, Alcorcón, Spain.
Introduction: Suffering from chronic pain (CP) and coping with parenthood can be challenging for parental mental health. Pain can hinder the ability to deal with demands related to parenthood, which can negatively affect their psychological well-being because of unmet caregiving expectations.
Methods: Considering the limited amount of research regarding the mental health of parents with CP, the study's main aim was to test a predictive model based on previous scientific literature, using structural equation analysis, in which parental competence and parental guilt partially mediate the relationship between parental stress and depression.
Impact of dysregulated microbiota-derived C18 polyunsaturated fatty acid metabolites on arthritis severity in mice with collagen-induced arthritis.
Front Immunol
January 2025
Department of Rheumatology and Clinical Immunology, Kobe University Graduate School of Medicine, Kobe, Japan.
Objective: We aimed to evaluate microbiome and microbiota-derived C18 dietary polyunsaturated fatty acids (PUFAs), such as conjugated linoleic acid (CLA), and to investigate their differences that correlate with arthritis severity in collagen-induced arthritis (CIA) mice.
Methods: On day 84 after induction, during the chronic phase of arthritis, cecal samples were analyzed using 16S rRNA sequencing, and plasma and cecal digesta were evaluated using liquid chromatography-tandem mass spectrometry. Differences in microbial composition between 10 control (Ctrl) and 29 CIA mice or between the mild and severe subgroups based on arthritis scores were identified.
Correlation between night sweats and season fluctuation in China.
Front Public Health
January 2025
Shandong Academy of Chinese Medicine, Jinan, China.
Background: Night sweats are a condition in which an individual sweats excessively during sleep without awareness, and stops when they wake up. Prolonged episodes of night sweats might result in the depletion of trace elements and nutrients, affecting the growth and development of children.
Purpose: To investigate the relationship between sweat nights and season.
Assessment of heart rate deceleration capacity, heart rate deceleration runs, heart rate acceleration capacity, and lipoprotein-related phospholipase A2 as predictors in individuals with dementia.
Front Neurol
January 2025
Department of Neurology, The Affiliated Taizhou People's Hospital of Nanjing Medical University, Taizhou School of Clinical Medicine, Nanjing Medical University, Taizhou, Jiangsu, China.
Background: Autonomic dysfunction plays an essential role in dementia, however, it is not known whether electrocardiogram autonomic dysfunction-related indicators are associated with the severity of dementia. In this study, we attempted to investigate whether these indicators are correlated in patients with vascular dementia and Alzheimer's disease compared with normal health individuals. For this purpose, we measured and analyzed the predictive value of heart rate deceleration capacity (DC), heart rate deceleration runs (DRs), heart rate acceleration capacity (AC) along with the plasma levels of lipoprotein-associated phospholipase A2 (Lp-PLA2).
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!