In an effort to generate a desired expression construct for making heart-specific expression transgenic zebrafish, a Tol2 plasmid, which can drive EGFP reporter gene specifically expressed in the heart, was modified using subcloning technology. An IRES fragment bearing multiple cloning site (MCS) was amplified directly from pIRES2-EGFP plasmid and was inserted between the CMLC2 promoter and EGFP fragment of the pDestTol2CG vector. This recombinant expression plasmid pTol2-CMLC2-IRES-EGFP can drive any interested gene specifically expressed in the zebrafish heart along with EGFP reporter gene. To test the effectiveness of this new expression plasmid, we constructed pTol2-CMLC2-RED-IRES-EGFP plasmid by inserting another reporter gene DsRed-Monome into MCS downstream of the CMLC2 promoter and injected this transgenic recombinant plasmid into one-cell stage embryos of zebrafish. Under fluorescence microscope, both the red fluorescence and the green fluorescence produced by pTol2-CMLC2-RED-IRES-EGFP were detected specifically in the heart tissue in the same expression pattern. This novel expression construct pTol2-CMLC2-IRES-EGFP will become an important tool for our research on identifying heart development candidate genes' function using zebrafish as a model.
Animals ; Animals, Genetically Modified ; genetics ; growth & development ; DNA Transposable Elements ; genetics ; Genes, Reporter ; genetics ; Genetic Vectors ; biosynthesis ; genetics ; Green Fluorescent Proteins ; genetics ; Myocardium ; metabolism ; Plasmids ; genetics ; Transfection ; Transgenes ; Transposases ; genetics ; Zebrafish ; genetics ; Zebrafish Proteins ; genetics ; metabolism

Objective:To explore the difference of blood pressure compliance rate in patients with continuous ambulatory peritoneal dialysis (CAPD) in the internet of things (IoT) follow-up and conventional care.Methods:CAPD patients from 3 peritoneal dialysis centers from May 2019 to October 2019 were included in this retrospective cohort study. They were divided into IoT group and conventional care group according to the way of follow-up. The difference in blood pressure compliance rate during 1 year of follow-up between the two groups was observed. The primary outcome was defined as the proportion of patients with blood pressure compliance rate≥85%.Results:A total of 75 patients were included in this study, in during 32 patients in IoT group and 43 patients in conventional care group. The comparison of baseline data between the two groups showed that the dialysis age of patients in IoT group was shorter ( P<0.01). After a median of 9(9, 12) months follow-up, the median blood pressure compliance rate was 85.2% (65.2%, 95.1%), and 25 patients (65.6%) in IoT group had met the target of blood pressure compliance rate≥85%, which was significantly higher than that in the conventional care group (17 cases, 39.5%) ( χ2=4.996, P=0.025). The cumulative probability of the target of blood pressure compliance rate≥85% was 97%, 90%, 90% and 52%, respectively in IoT group, while 95%, 86%, 55% and 34%, respectively in conventional care group after 3, 6, 9 and 12 months of follow-up, and the different between the two groups was significant (Log-rank χ2=4.774, P=0.029). Adjusted for age, sex and dialysis age, the multivariate Cox proportional risk regression model showed that serum creatinine level(for every 1 μmol/L increase, HR=1.002, 95% CI 1.000-1.003, P=0.033), follow-up mode (IoT follow-up vs conventional care, HR=0.023, 95% CI 0.003-0.210, P=0.001), follow-up times (for each additional time, HR=0.879, 95% CI 0.823-0.939, P<0.001) and the rate of weight compliance (for each increase of 1%, HR=0.964, 95% CI 0.939-0.991, P=0.008) was the independent influencing factors for the blood pressure compliance rate<85%. The results of subgroup analysis showed that patients with shorter dialysis age (<10 months) and in the centers where the nurses finished the PD follow-up work as part-time job had better blood pressure control in IoT follow-up. Conclusions:IoT follow-up is helpful to improve CAPD patients' blood pressure compliance rate. Elevated serum creatinine level at baseline is the independent risk factor associated with poor blood pressure compliance. However, IoT follow-up, more follow-up times and the elevated rate of weight compliance are the protective factors for blood pressure compliance. IoT follow-up mode is more recommended for patients with short dialysis age and for dialysis centers where most of the nurses are part-time.