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OBJECTIVE@#To explore the genetic basis of cerebral palsy (CP).@*METHODS@#A pair of twins with cerebral palsy and different phenotypes were subjected to whole genome sequencing, and other 8 children with CP were subjected to whole exome sequencing. Genetic variations were screened by a self-designed filtration process in order to explore the CP-related biological pathways and genes.@*RESULTS@#Three biological pathways related to CP were identified, which included axon guiding, transmission across chemical synapses and protein-protein interactions at synapses, and 25 susceptibility genes for CP were identified.@*CONCLUSION@#The molecular mechanism of CP has been explored, which may provide clues for development of new treatment for CP.
Subject(s)
Child , Humans , Cerebral Palsy , Genetics , Genetic Testing , Phenotype , Exome Sequencing , Whole Genome SequencingABSTRACT
AIM:To investigate the inhibitory effect of aliskiren on the angiogenesis of human umbilical vein endothelial cells ( HUVECs) induced by lipopolysaccharide ( LPS) and to explore its possible mechanism.METHODS:HUVECs were cultured and randomly divided into blank group and renin group.The levels of tumor necrosis factor-α(TNF-α) and intercellular adhesion molecule-1 (ICAM-1) in the culture supernatant were detected by ELISA.The protein levels of Toll-like receptor 4 ( TLR4) and ICAM-1 in the HUVECs were determined by Western blot.HUVECs were cul-tured and randomly divided into control group, LPS group, low-dose (1μmol/L) aliskiren group, middle-dose (10μmol/L) aliskiren group and high-dose (100 μmol/L) aliskiren group.The proliferation of HUVECs was detected by MTT and BrdU assays.The mobility of HUVECs was measured by Transwell assay.The formation of the vessels was judged by ob-serving the formation of the luminal structure by HUVECs in Matrigel.The levels of TNF-α, ICAM-1 and monocyte chemo-tactic protein 1 ( MCP-1) in the culture supernatant were measured by ELISA.The expression of renin, TLR4, matrix me-talloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) at mRNA and protein levels in the HUVECs was determined by RT-PCR and Western blot.RESULTS:Renin stimulated the expression of inflammatory factors and TLR4 in the HUVECs.Aliskiren inhibited the growth, migration and angiogenesis of HUVECs in a dose-dependent manner, de-creased the levels of TNF-α, ICAM-1 and MCP-1 and the expression of renin, MMP-2 and MMP-9, and inhibited TLR4 expression (P<0.05).CONCLUSION:Aliskiren inhibits LPS-induced angiogenesis of HUVECs, which may be related to the down-regulation of renin expression, the inhibition of TLR4-mediated inflammatory reaction, and the formation of MMP-9 and MMP-2.
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AIM:To investigate the inhibitory effect of catalpol on inflammation in EA .hy926 cells induced by advanced glycation end products ( AGEs) and to explore its antioxidant mechanisms .METHODS:Human endothelial cell line EA.hy926 was cultured and randomly divided into control group , catalpol (0.5 mmol/L) group, AGEs group, high-dose (0.5 mmol/L) catalpol +AGEs group, middle-dose (0.25 mmol/L) catalpol +AGEs group and low-dose (0.05 mmol/L) catalpol+AGEs group.Intracellular reative oxygen species ( ROS) production was detected by laser scanning confocal microscopy.The levels of monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-α(TNF-α) and vas-cular cell adhesion molecule-1 (VCAM-1) in culture supernatant were detected by commercial ELISA kits .The expression of MCP-1, TNF-α, VCAM-1 and receptor for advanced glycation end products (RAGE) in the EA.hy926 cells were detec-ted by Western blot.RESULTS:In high-dose catalpol+AGEs and middle-dose catalpol+AGEs groups, the generation of ROS was decreased significantly .The levels of MCP-1, TNF-αand VCAM-1, and protein expression of MCP-1, TNF-αand VCAM-1 were significantly lower .The expression of RAGE protein in EA .hy926 cells were significantly inhibited ( P<0.05).CONCLUSION:Catalpol effectively inhibits the AGEs-induced oxidative stress and inflammation in EA .hy926 cells, which may be associated with a decrease in the expression of RAGE .
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AIM:To investigate the protective effect of catalpol on Goto-kakizaki (GK) rat aorta and to ex-plore its antioxidant mechanisms.METHODS:Six-month-old GK rats (n=45) were randomly divided into diabetic model group, metformin (100 mg· kg-1· d-1) group, and high-dose (100 mg· kg-1· d-1), medium-dose (50 mg· kg-1· d-1) and low-dose (10 mg· kg-1· d-1) catalpol groups.The healthy male Wistar rats (n=10) were used as control group.The rats in control and model groups were given a same volume of saline .All reagents were administered by oral ga-vage for 12 weeks.Blood glucose and lipids were detected by an automatic biochemical analyzer .Serum reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and total antioxidant capacity (T-AOC) levels were detected by commercial kits .The expression of nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in the thoracic aorta was determined by Western blotting .The pathological changes of the thoracic aorta were observed by HE staining.The ultrastructural changes of the thoracic aorta were observed under electron microscope .RESULTS:Af-ter catalpol treatment , the levels of blood glucose and blood lipids were decreased significantly , and serum levels of ROS and MDA were significantly decreased , but the activity of SOD and T-AOC were significantly enhanced .The protein ex-pression of Nrf2 and HO-1 in the thoracic aorta were significantly increased , the thoracic aortic lesions indicated by HE staining significantly reduced , and the thoracic aortic damage under ultrastructural observation was attenuated slightly . CONCLUSION:Catalpol effectively protects GK rat thoracic aorta , which may be associated with decreasing blood lipids , reducing oxidative stress and activating Nrf 2/ARE/HO-1 signaling pathways.