Objective To explore the impacts and potential mechanisms of MitoQ on isoflurane-induced injury of primary cultured hippocampal neurons in newborn rats.Methods Fifteen healthy SPF Sprague-Dawley rats of both sex were randomly divided into three groups (n =5 each)u-sing a random number table:control group (group C),multiple exposures to isoflurane anesthesia group (group I)and multiple exposures to isoflurane anesthesia+MitoQ group (group IM).On post-natal days 7,14 and 21,1.5% isoflurane was inhaled for 2 h in group I.MitoQ was intraperitoneally administered in a volume of 0.4 ml/kg before isoflurane anesthesia in group IM,while a mixture of oxygen and air was inhaled instead of isoflurane in group C.HE staining was carried out on postnatal day 28 to observe the morphological changes in hippocampal CA1 region of rat neural cell structures. Hippocampal neuron cells were dissected from clean Sprague-Dawley rats born in 24 h.After primary culture for seven days,MTT assay and TUNEL assay was respectively performed to measure the cell viability and apoptosis of hippocampal neurons.The malondialdehyde (MDA)content and superoxide dismutase (SOD)activity were detected respectively using the thiobarbituric method and xanthinoxi-dase method.Mitochondrial membrane potential (MMP)was measured by rhodamine 123 staining, intracellular levels of reactive oxygen species (ROS)were tested by DCFH-DA staining.Western blot was used to analyze the protein levels of Bax,Bcl-2 and caspase-3.Results Compared with group C, group I decreased the number of neural cells and the cell survival rate;the apoptotic rate was signifi-cantly increased;MDA contents and ROS production were significantly increased;SOD activity and MMP level were significantly decreased;the expression of Bax and caspase-3 were significantly in-creased,while the expression of Bcl-2 was significantly decreased (P < 0.05 ).Compared with the group I,the damaged neural cells were decreased,the cell survival rate was significantly increased, the apoptotic rate was significantly decreased in group IM;MDA contents and ROS production were significantly decreased;SOD activity and MMP level were significantly increased;the expression of Bax and caspase-3 were significantly decreased,while the expression of Bcl-2 was significantly in-creased (P < 0.05 ).Conclusion Antioxidant MitoQ attenuates isoflurane-induced neuron damage, which may be associated with the inhibition on oxidative stress and mitochondrial dysfunction.

ObjectiveTo clone coumarate-3-hydroxylase gene （C3H） from Angelica sinensis， and analyze the correlation between its bioinformatics， expression patterns and content of ferulic acid， and to explore the functions of ASC3H. MethodReal-time polymerase chain reaction （Real-time PCR） was used to clone the full-length cDNA of ASC3H based on the transcriptome dataset of A. sinensis， and the bioinformatics analysis of the gene sequence was carried out. Real-time PCR and high performance liquid chromatography （HPLC） were used to determine relative expression of ASC3H and content of ferulic acid in different root tissues of A. sinensis （periderm， cortex and stele）. ResultThe open reading frame （ORF） of ASC3H （GenBank accession number： MN2550298） was 1 530 bp， encoding 509 amino acids， with a theoretical molecular weight of 57.86 kDa and an isoelectric point of 8.36. It was a hydrophilic protein that was located in the chloroplast with multiple phosphorylation sites and a transmembrane region， and contained a conserved domain CGYDWPKGYGPIINVW_P450 （383-399 aa） in cytochrome P450. Multiple amino acid sequence alignment analysis showed that ASC3H had high similarity with C3H from other plants， especially Ammi majus in Umbelliferae. The Real-time PCR revealed that ASC3H had different expressions in periderm， cortex and stele tissues of A. sinensis roots. It was found from HPLC that the cortex tissues had the highest content of ferulic acid， and the stele tissues had the lowest. ConclusionASC3H was successfully cloned from A. sinensis， and its sequence characteristics were understood more clearly， suggesting that ASC3H might be involved in the ferulic acid biosynthesis pathway of A. sinensis. This paper provided a basis for further studying the functions of the gene and exploring the biosynthesis and regulation mechanism of ferulic acid in A. sinensis， while laying the foundation for the genetic improvement of A. sinensis．