ABSTRACT
Objective To analyze the effect of the identification and evaluation of Escherichia (E.) coli and Shigella,based on the upstream flanking sequences of CRISPR1.Methods Both CRISPR and cas sequences were obtained through the BLAST with repeating sequences against the publicly complete genome in GenBank that related to E.coli and Shigella.Clustal X was used to perform multi-sequences alignment of the flanking sequences.PCR method was used to amplify the upstream flanking sequences of CRISPR1 in order to appraise the effect of identification and evaluation of upstream flanking sequences on E.coli and Shigella,which were based on the upstream flanking sequences of CRISPR1.Results The results showed that 73.4% of the strains containing the I-E CRISPR/Cas that belonged to the phylogroups A,B1,D while 8.4% strains carried the I-F CRISPR/Cas.Another 17.2% of the strains owned CRISPR3-4 (non-CRISPR/Cas) only belonged to the phylogroups B2.All the Shigella strains carried I-E CRISPR/Cas.More than 99% of similarity the CRISPR1 upstream-flanking sequences was seen in E.coli (except B2) and Shigella and E.coli (B2).Both sensitivity and specificity were greater than 91% after PCR amplification in the region to identify the E.coli and Shigella.Conclusion The upstream of CRISPR1 could achieve a preliminary identification effect on E.coli and Shigella.
ABSTRACT
Objective: To analyze the effect of the identification and evaluation of Escherichia (E.) coli and Shigella, based on the upstream flanking sequences of CRISPR1. Methods: Both CRISPR and cas sequences were obtained through the BLAST with repeating sequences against the publicly complete genome in GenBank that related to E. coli and Shigella. Clustal X was used to perform multi-sequences alignment of the flanking sequences. PCR method was used to amplify the upstream flanking sequences of CRISPR1 in order to appraise the effect of identification and evaluation of upstream flanking sequences on E. coli and Shigella, which were based on the upstream flanking sequences of CRISPR1. Results: The results showed that 73.4% of the strains containing the I-E CRISPR/Cas that belonged to the phylogroups A, B1, D while 8.4% strains carried the I-F CRISPR/Cas. Another 17.2% of the strains owned CRISPR3-4 (non-CRISPR/Cas) only belonged to the phylogroups B2. All the Shigella strains carried I-E CRISPR/Cas. More than 99% of similarity the CRISPR1 upstream-flanking sequences was seen in E. coli (except B2) and Shigella and E. coli (B2). Both sensitivity and specificity were greater than 91% after PCR amplification in the region to identify the E.coli and Shigella. Conclusion: The upstream of CRISPR1 could achieve a preliminary identification effect on E.coli and Shigella.
Subject(s)
Humans , Clustered Regularly Interspaced Short Palindromic Repeats/genetics , DNA, Bacterial/genetics , Escherichia coli/isolation & purification , Genotype , Molecular Sequence Data , Sequence Analysis, DNA , Shigella/isolation & purificationABSTRACT
Objective:To investgate the ease effects of local injection of ursolic acid both on orthodontic tooth movement distance and tooth root resorption in the rats, and to clarify its mechanism.Methods:Ninety-six male Wistar rats were used to establish models of orthodontic tooth movement.After the successful establishment of models,all the model rats were randomly divided into 0 (control group),0.5,1.0 and 2.0 mmol·L-1 ursolic acid groups (n=24).The rats were locally injected with different doses of ursolic acid into the palatal submucosal area adjacent to the right upper first molar ,once per 3 d,each time 50μL.The rats were respectively sacrificed on the days 1,3,5,7,10,14,21 and 28,then the distances of tooth movement were measured.HE staining was used to observe the morphologic changes of the root tissue of rats.Results:The tooth movement distances of rats in control,0.5,1.0,and 2.0 mmol· L-1 ursolic acid groups were increased with the prolongation of time (P0.05),but at the other time points,the tooth movement distances of rats had statistical significance (P<0.01).The tooth movement distances of rats had statistical significance between 0.5,1.0 and 2.0 mmol·L-1 ursolic acid groups 5,7,10,14,21 and 28 d after forcing (P<0.05 or P<0.01).The morphological results showed that the resorption pits were found on the surface of tooth root with the prolongation of loading time;in addition,with the increasing of the doses of ursolic acid, the absorption of root surface was relieved. Conclusion:Local injection of ursolic acid at the doses of 0.5,1.0 and 2.0 mmol·L-1 can reduce the distance of orthodontic tooth movement in the rats, and the distance of tooth movement in the rats is reduced with the increasing of its doses,and ursolic acid has the ease effect on the orthodoutic tooth root resorption.