Objective To study the genome molecular characteristics of Getah virus(DY0824)which isolated in Shandong province,2008 by molecular biology methods.Methods Reverse transcriptasepolymerase chain reaction(RT-PCR)was used to amplify the structural gene and 3'UTR fragments then the RT-PCR products were inserted into PGEM-T easy to be sequenced.Computer software was used to analyze the nucleotide and deduced amino acid sequence,and draw phylogenetic trees,including Clustal X1.83 and MegaAlign and Mega4.Results The capsid protein of DY0824 consists of 804 nucleotides,encoding 268 amino acids and the full-length of E2 protein is 1266 nucleotides,encoding 422 amino acids.The nucleotide homology of the capsid protein and the E2 protein with other strains were 95.4%-99.9%and 94.8%-99.5%,and the amino acid were 97.4%-100%and 97.6%-100%.The 3'UTR of the virus include 401 nucleotides and there are three repeat sequence elements.Conclusion Compared with the prototype virus,the Getah virus isolated in Shandong province had 7 amino acid differences in capsid protein genes and 10 amino acid differences in E protein genes.The 3'UTR region had multi-nucleotide changes.

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective To investigate the lead and cadmium contents in different sampling sites from full term neonatal placenta and to explore the role of placental sample in the evaluation of intrauterine heavy metals exposure.Methods The placentas from 30 healthy full term neonates were collected from the West China Second Hospital of Sichuan University during May and June 2016.Each placenta fetal side was divided into the left and right parts with the umbilical vein in the umbilical cord cross-section as the 12 o'clock direction.The villus lobular tissue samples were taken from the 1/4 radius in left part (site A) and 3/4 radius in the right part (site B).The graphite furnace atomic absorption spectrometry was used to detect the lead and cadmium contents in the samples.The elements contents in the site A and B were performed the paired t-test and correlation analysis.Results The mean contents of lead and cadmium in dry weight sample at placental site A were 91.8 and 66.7μg/kg which at the site B were 88.9 and 64.8 μg/kg respectively.The lead and cadmium contents at these two sites presented the positive correlation,the coefficients were 0.98 and 0.97 respectively,whereas the difference in contents between the tissues from different placental sites had no statistical significance.Conclusion The lead and cadmium contents of villus lobular tissue in the central part of placenta fetal side (1/4-3/4 radius area) are basically consistent,which is a reliable indicator for evaluating the intrauterine heavy metals exposure.

【Objective】 To evaluate the potential of miR-30a-5p as a novel indicator of acute myocardial infarction (AMI) and the underlying molecular mechanisms. 【Methods】 AMI-associated microRNAs (miRNAs) and mRNA microarray datasets were downloaded from the GEO database. Real-time fluorescence quantitative PCR (qRT-PCR) technique was used to detect the level of miRNAs in serum samples; automatic biochemistry was used to detect other biochemical indicators. Receiver operating characteristic (ROC) curve analysis and Spearson correlation analysis were performed to assess the value of miR-30a-5p as a diagnostic AMI marker. The target genes of miR-30a-5p were predicted with the R language multiMiR package, and the protein interaction network was constructed by the STRING database. The results were imported into Cytoscape 3.7.1 software to screen the pivotal genes of the network. The R language clusterProfiler package performed KEGG and GO analyses on the hub genes to explore the clinical significance of miR-30a-5p in AMI and its potential molecular mechanisms. 【Results】 Compared with the control group, miR-30a-5p was upregulated significantly in the serum of patients in the AMI group (P<0.05); miR-30a-5p was positively correlated with the levels of CK-MB, CK, TnT, proBNP and CRP (rs=0.489, P<0.001; rs=0.347, P<0.001; rs=0.545, P<0.001; rs=0.533, P<0.001; rs=0.206, P<0.05). The area under the ROC curve was 0.862, with sensitivity of 84.4% and specificity of 74.2%. A total of 780 differentially expressed genes (DEGs) were obtained from the two datasets. A total of 1 061 target genes of miR-30a-5p and 61 common genes were identified by taking the intersection set. Among the central genes of PPI, BCL6, FOSL2, JDP2, LYN, PDE4D, SOCS3 and SOX4 scored high and were closely associated with the occurrence of AMI. KEGG and GO enrichment analyses showed that miR-30a-5p might regulate JAK-STAT, NF-kappaB and Wnt signaling pathways, which were involved in inflammatory response, apoptosis, autophagy and post-infarction remodeling, and thus participated in the process of AMI. 【Conclusion】 Serum miR-30a-5p is up-regulated in the early stage of AMI, and the study on miR-30a-5p and its regulatory pathways can help with the diagnosis and treatment of myocardial infarction.

OBJECTIVE: Pseudostellaria heterophylla has been paid more attention in recent years, mainly as a medicine food homology plant. The content determination of P. heterophylla is not specified in the Chinese Pharmacopoeia (version 2020). The environmental conditions in different production areas could exert an influence on the quality of P. heterophylla. The purpose of this study is to discriminate P. heterophylla collected from different geographical origins of China. METHODS: In this study, the content of polysaccharide in 28 batches of P. heterophylla was determined using phenol-sulfuric acid. HPLC fingerprints were established under optimised HPLC-PDA methods. Subsequently, the similarity analysis (SA) and the quantification of heterophyllin B were analyzed. The metabolites of P. heterophylla were identified and evaluated using UHPLC-Q Exactive HF orbitrap MS system. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), hierarchical cluster analysis (HCA) and orthogonal PLS-DA (OPLS-DA) were performed based on all peak areas. RESULTS: The polysaccharide content in Guizhou and Jiangsu was higher than that of other production areas, which varied significant from different origins. While the content of heterophyllin B in Anhui and Jiangsu was high. The correlation coefficients of HPLC fingerprints for 28 batches samples ranged from 0.877 to 0.990, and the characteristic map can be used to identify and evaluate the quality of P. heterophylla. The samples from Fujian, Guizhou, Jiangsu provinces can be relatively separated using multivariate statistical analysis including PCA, PLS-DA, HCA, OPLS-DA, indicating that their metabolic compositions were significantly different. Ultimately, a total of 15 metabolites which were filtrated by a VIP-value > 1 and a P-value < 0.05 associated with the separation of different origins were identified. CONCLUSION HPLC fingerprint was established to evaluate the quality and authenticity of P. heterophylla. The present work showed that the difference of geographic distributions had an influence on the internal chemical compositions. A sensitive and rapid untargeted metabolomics approach by UHPLC-Q Exactive HF orbitrap MS was utilized to evaluate P. heterophylla from different origins in China for the first time. Overall, this study provides insights to metabolomics of P. heterophylla and supplies important reference values for the development of functional foods.