ABSTRACT
Objective:To clone uridine diphosphate （UDP）-glucose dehydrogenase （<italic>UGDH</italic>） gene of <italic>Glycyrrhiza uralensis</italic> and analyze its bioinformatics and expression. Method:Total RNA was extracted from roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， the complementary deoxyribonucleic acid （cDNA） sequence of <italic>GuUGDH</italic>1 gene （Gu was short for <italic>G. uralensis</italic>） was cloned by reverse transcription polymerase chain reaction （RT-PCR）， then sequencing and bioinformatic analysis were performed， and the specificity of the tissue was analyzed by real-time fluorescence quantitative PCR （Real-time PCR）. Result:The open reading frame（ORF）of <italic>GuUGDH</italic>1 gene was 1 443 bp in length and encoded 480 amino acid residues （GenBank accession number of MT968993）. Bioinformatics analysis showed that GuUGDH1 was a stable acidic hydrophilic protein with a relative molecular weight of 53.056 kDa， an isoelectric point of 5.89， no signal peptide and no transmembrane helix， and all of them were outside the membrane. There were three typical conserved domains， which belonged to the UDP-glucose/guanosine diphosphate （GDP）-mannose dehydrogenase family. Phylogenetic analysis showed that the <italic>GuUGDH</italic>1 gene was closely related to <italic>Glycine max</italic> and <italic>Spatholobus suberectus</italic>. The results of Real-time PCR showed that the expression of <italic>GuUGDH</italic>1 gene could be detected in the roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， and the expression level in the roots was significantly higher than that in the stems and leaves. Conclusion:In this study， the <italic>UGDH</italic>1 gene of <italic>G. uralensis</italic> was cloned and its protein sequence characteristics were systematically analyzed， which can provide theoretical basis for further research on the catalytic function of UGDH1 protein.
ABSTRACT
Objective:To identify 24 <italic>Rana</italic> species such as <italic>Rana dybowskii</italic> by mitochondrial cytochrome C oxidase subunit I （<italic>CO</italic>Ⅰ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of <italic>Rana</italic> species as well as the discovery of new species. Method:<italic>R. dybowskii</italic>， <italic>R. chensinensis</italic>， <italic>R. amurensis</italic>， <italic>R. culaiensi</italic>s， and <italic>R. huanrenesis</italic>， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification<italic> </italic>and sequencing. A total of 50 <italic>CO</italic>Ⅰ gene sequences were obtained. Then 163 <italic>CO</italic>Ⅰ gene sequences for 24 species of <italic>Rana</italic> and one <italic>CO</italic>Ⅰ gene sequence for <italic>Pelophylax</italic>，<italic> Odorrana</italic>， <italic>Nidirana</italic>， <italic>Hylarana</italic>， and <italic>Amolops</italic> were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of <italic>CO</italic>Ⅰ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis. Result:The <italic>CO</italic>Ⅰ gene sequences of 24<italic> Rana</italic> species including <italic>R. dybowskii</italic> were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> were clustered into one branch and some <italic>R. dybowskii</italic> and <italic>R. uenoi</italic> into one branch. There were two separate branches for <italic>R. chensinensis</italic> and the other species were all clustered independently. Conclusion:<italic>CO</italic>Ⅰ-based DNA barcoding enabled the identification of 24 species of <italic>Rana</italic> including <italic>R.dybowskii</italic>. The findings supported that <italic>R. sangzhiensis</italic>， <italic>R. zhengi</italic>， <italic>R. coreana</italic>， and <italic>R. kunyuensis</italic> were the same species. One branch of <italic>R. chensinensis </italic>might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that <italic>CO</italic>Ⅰ-based DNA barcoding allows not only the identification of 24 species of Rana including <italic>R. dybowskii </italic>but also the classification of ranidae species and the discovery of new species or subspecies.
ABSTRACT
OBJECTIVE@#To study the expression of microRNA-495-5p (miRNA-495-5p) in the serum of preterm infants with bronchopulmonary dysplasia (BPD) based on a bioinformatics analysis, and to provide a theoretical basis for further research on the association between miRNA-495-5p and BPD.@*METHODS@#A total of 40 preterm infants who were admitted to the neonatal intensive care unit from January 2015 to December 2016 were enrolled. Among these infants, 20 with early clinical manifestations of BPD were enrolled as the BPD group, and 20 without such manifestations were enrolled as the control group. Peripheral blood samples were collected. The miRNA microarray technique was used to screen out differentially expressed miRNAs in serum between the two groups. RT-PCR was used for validation of results. TargetScan, miRDB, and miRWalk databases were used to predict the target genes of miRNA-495-5p. The DAVID database was used to perform gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the target genes.@*RESULTS@#Compared with the control group, the BPD group had a significant increase in the expression of miRNA-495-5p in serum (P<0.05). A total of 117 target genes of miRNA-495-5p were predicted by the above three databases and they were involved in several molecular functions (including transcriptional regulatory activity, transcriptional activation activity, and transcription cofactor activity), biological processes (such as metabolic regulation, DNA-dependent transcriptional regulation, and vascular pattern), and cell components (including nucleoplasm, membrane components, and insoluble components) (P<0.05). As for signaling pathways, these genes were significantly enriched in the mTOR signaling pathway (P<0.05).@*CONCLUSIONS@#MiRNA-495-5p may be involved in the development and progression of BPD by regulating angiogenesis, stem cell differentiation, apoptosis, and autophagy, which provides clues for further research on the role and functional mechanism of miRNA-495-5p in BPD.
Subject(s)
Humans , Infant, Newborn , Bronchopulmonary Dysplasia , Computational Biology , Infant, Premature , MicroRNAs , Genetics , Transcription, GeneticABSTRACT
Eight triterpenes were isolated from the methanol extract of Galbanum by various chromatographic methods including silica gel, ODS opening column, recrystallization and semi-preparative HPLC. Their structures were determined by spectroscopic methods and physicochemical properties as 3β,19α,21α-trihydroxyl-12-en-28-oic acid (1), sumaresinolic acid (2), 3β,19α-dihydroxyl-12-en-28-oic acid (3), oleanolic acid (4), 3β,6β,19α-trihydroxyl-12-en-28-oic acid (5), 19α-hydroxy oleanonic acid (6), 6α-hydroxy oleanonic acid (7), and (11R,12R)-3α,6α-dihydroxy-epoxyolean-28α,13α-olide (8). Among them, compound 1 is a new compound, while compounds 2-8 were newly isolated from the Apiaceae family. The ability of compounds 1-8 to inhibit cholinesterase was determined with an improved Ellman method. Compound 1 showed strong inhibitory activity against butyrylcholinesterase. The molecular docking results indicated that Trp82, His438, Phe329 and Ala328 played an important role in the binding of compound 1 to butyrylcholinesterase.
ABSTRACT
As a traditional Chinese medicine in China, ginseng has a wide range of medicinal and health value. At present, the nutritional value of ginseng as a medicinal food has been a hotspot in studies. Intestinal flora plays an important role in the organism, which has been confirmed by many researchers. In order to find out the effect of long-term intake of ginseng extracts on the gut microbiota structure of rats, MiSeq sequencing platform was applied in macro gene sequencing of cecal contents in the long-term use of ginseng extracts modelin rats. According to the findings, after long-term administration with ginseng extracts, probiotics such as Bifidobacterium, Lactobacillus, Allobaculum and Clostridium, in the intestinal flora of rats were significantly increased, suggesting that long-term intake of ginseng extracts could facilitate the growth of probiotics. Meanwhile, some pathogenic bacteria, such as Butyricimonas, Parabacteroides, Alistipes, Helicobacter, were significantly down-regulated, indicating that long-term intake of ginseng extracts may have a positive effect in inhibiting the colonization of pathogenic bacteria. In conclusion, this study provided an important basis for the research on the effect of long-term use of ginseng extracts on the intestinal flora of rats.
Subject(s)
Animals , Rats , Bacteria , Classification , China , Gastrointestinal Microbiome , Panax , Chemistry , Plant Extracts , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To predict the target genes of rno-microRNA-296-5p (miR-296) using bioinformatics software and databases, and to provide a theoretical basis for further studies of biological effects of miR-296 in fetal lung development.</p><p><b>METHODS</b>PubMed and Google were used to search for all reported literature on miR-296. The miRBase database was used to determine the sequence and evolutionary conservatism of miR-296. The TargetScans database was used to predict the target genes of miR-296. The DAVID Bioinformatics Resources 6.8 database was used for the functional enrichment analysis of the target genes. The KEGG database was used to analyze the signaling pathways of target genes.</p><p><b>RESULTS</b>miR-296 was reported to play important roles in many biological processes and have a high degree of sequence conservation among species. The target genes of miR-296 were involved in biological processes, cell components, and molecular function. Those target genes were significantly enriched in the mitogen-activated protein kinase signaling pathway, Wnt signaling pathway, and transforming growth factor-β signaling pathway (p<0.05).</p><p><b>CONCLUSIONS</b>The bioinformatics analysis of the target genes of miR-296 provides a basis for studying biological effects and mechanism of action of miR-296 in lung development.</p>