Objective:To systematically evaluate the efficacy of epinephrine in preventing post-endoscopic retrograde cholangiopancreatography pancreatitis (PEP).Methods:Randomized controlled trials (RCTs) on epinephrine for preventing PEP from inception to October 10, 2020 were searched in databases including PubMed, Embase, The Cochrane Library, Web of Science, VIP Information Network, China National Knowledge Infrastructure,WanFang Data,and clinical trial registration platforms including ClinicalTrials.gov,WHO International Clinical Trial Registration Platform. Literature was screened independently by two reviewers, data were extracted and the risk of bias of included studies were assessed. The meta-analysis was performed by RevMan 5.3.Results:A total of 410 papers were retrieved and 8 RCTs involving 4 208 patients were included. The results of meta-analysis showed that compared with the saline group, the epinephrine could reduce the incidence of PEP ( RR=0.29,95% CI:0.16-0.50, P<0.001). There were no significant differences in the therapeutic effect between group epinephrine and group indomethacin ( RR=0.17,95% CI:0.02-1.39, P=0.100) or group indomethacin combined with epinephrine and group indomethacin ( RR=1.15,95% CI:0.61-2.16, P=0.670). Conclusion:Local spraying of epinephrine on the duodenal papilla can reduce the incidence of PEP compared with normal saline. But the epinephrine or combination of indomethacin and epinephrine fails to reveal any benefit over indomethacin alone in preventing PEP.

OBJECTIVETo construct a luciferase reporter vector containing the response element of transcription protein AP2α for screening the effect of bone morphogenetic proteins (BMPs) on the transcriptional activity of AP2α.

METHODSFour tandem-linked response elements of AP2α were cloned to the pBGLuc luciferase reporter gene plasmid, which was digested with Bam HI and Mlu I to construct pBGLuc-AP2α-RE vector. The recombinant adenovirus Ad-AP2α and its dominant negative mutant Ad-dnAP2α were used to infect mouse mesenchymal stem cells C3H10; the changes in cellular AP2α mRNA and protein expressions were detected by real-time PCR and Western blotting, and electrophoretic mobility shift assay (EMSA) was carried out to assess the DNA-binding ability of AP2α. C3H10 cells were transfected with pBGLuc-AP2α-RE vector, and AP2α transcriptional activity was measured using luciferase reporter gene assay. In pBGLuc-AP2α-RE vector-transfected C3H10 cells infected with Ad-BMPs, luciferase reporter gene assay was performed to screen the effect of BMPs on AP2α transcriptional activity.

RESULTSThe results of PCR, enzyme digestion and sequencing all confirmed correct cloning of AP2α-RE into pBGLuc-AP2α-RE luciferase reporter vector, and Ad-AP2α infection significantly increased AP2α expression and its DNA binding ability. The dominant negative mutants expressed the corresponding mutants, and EMSA results showed that Ad-dnAP2α-δbHLH significantly lowered while Ad-dnAP2α-δTAD enhanced the DNA-binding ability of AP2α. AP2α over-expression promoted AP2α transcriptional activity, which was suppressed by the two dominant negative mutants. AP2α transcriptional activity increased in the cells infected with the recombinant adenovirus BMPs, especially in cells with BMP9 infection.

CONCLUSIONSThe luciferase reporter vector containing the response element of AP2α we constructed allows detection of AP2α transcriptional activity. BMP9 can significantly enhance AP2α transcriptional activity.

Adenoviridae ; Animals ; Bone Morphogenetic Proteins ; genetics ; metabolism ; Genes, Reporter ; Genetic Vectors ; Growth Differentiation Factor 2 ; genetics ; metabolism ; Luciferases ; genetics ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Osteogenesis ; Protein Binding ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Response Elements ; Transcription Factor AP-2 ; genetics ; metabolism ; Transcriptional Activation ; Transfection