CCNE1, CDK2, and CREB1 are directly involved in IGF-1 stimulated crypt cell proliferation.

<p>(A) immunoblotting analyses of CCNE1, CDK2, and CREB1 protein in mouse small intestinal crypt cells with or without IGF-1 treatment for 24 h. Data were normalized to β-actin. Values are means ± SEM run in triplicates, asterisks indicate significant differences between control and IGF-1 treated cells. (B) CCNE1, CDK2, and CREB1 proteins were knocked-down by siRNA in IEC-6 cells before treated with IGF-1. Cell proliferation was measured 24 h after IGF-1 treatment. Values are means ± SEM run in triplicates, letters indicate significant differences between time points.</p

Mouse small intestinal crypt cells express PCNA.

<p>PCNA expression was analyzed by immunoblotting. 50 µg total proteins from extracted mouse small intestinal crypt cells and cultured rat intestinal crypt cell line IEC-6 were loaded. PCNA was expressed in both cell types and with significantly higher abundance in mouse small intestinal crypt cells (<i>P</i><0.01) than in IEC-6 cells. β-actin served as a loading control. Values are means ± SEM run in triplicates.</p

IGF-1 stimulates mouse small intestinal crypt cell proliferation.

<p>Crypt cells in 96-well plate were treated with 20 ng/ml IGF-1; the control group was grown in the crypt cell growth medium alone. After treatment for 24 h, rate of cell proliferation was measured by Brdu incorporation assay. IGF-1 significantly stimulated proliferation of the crypt cells (<i>P</i><0.05). Values are means ± SEM run in triplicates; asterisks indicate significant differences between IGF-1 treatment and control groups.</p

Forty-four miRNAs were significantly correlated with mouse crypt cell proliferation upon IGF-1 treatment.

<p>Hierarchical clustering analysis was performed using Euclidian distance. Each row represents relative levels of expression for a significantly regulated single microRNA and each column represents the relative expression level of a single replicate relative to control (<i>P</i><0.01). Colors on the figure represent the scaled fold-change between samples within a gene. Control group was set to 0.</p

CCNE1, CDK2, and CREB1 are direct targets of miR-103 in mouse small intestinal crypt cells during IGF-1 stimulation.

<p>(A) sequence alignment between miR-103 seed region and the seed matches on CCNE1, CDK2, and CREB1 mRNA 3′UTR region. The analyses were performed using the miRBase target database. The line indicates conserved seed match (A–T, C–G) in human and mouse, whereas the dot indicates seed match in one of the two species. (B, C) miR-103 directly binds and represses CCNE1, CDK2, and CREB1 mRNA through 3′UTR. pGL3-control luciferase vectors containing the mRNA 3′UTR of respective genes of human (B) or mouse (C) origin were co-transfected with the indicated amount of miR-103 mimic in HEK293 cells, and luciferase activity was analyzed 24 h post-transfection. The structurally unrelated miRNA cel-miR-67 served as negtive control. (D) CCNE1, CDK2, and CREB1 protein expression were examined in IEC-6 cells after transfection with 50 nM miR-103 inhibitor. Scramble (scr) miRNA transfected cells served as control. Values are means ± SEM run in triplicates, and letters indicate significant differences between treatment groups and control.</p

miR-103 is directly involved in IGF-1 stimulated crypt cell proliferation.

<p>IEC-6 cells in 96-well plate were transfected with 50 nM miR-103 mimic before treated with 20 ng/ml IGF-1. The control group was cultured in growth media or treated with IGF-1 alone. Cell proliferation was measured by BrdU incorporation assay. Values are means ± SEM run in triplicates. Letters indicate significant differences between IGF-1 treatment and control groups.</p

miR-103 expression analysis in IGF-1 stimulated mouse small intestinal crypt cells.

<p>(A) microrarray analysis of total miRNA expression in crypt cells after stimulation with IGF-1 for 24 h. The MA plot shows the averaged and background-subtracted fold change on a log2 scale (Y axis) and average expression intensity (X axis) of each miRNA on both channels for Cy-3 labeled control and Cy-5 labeled treated cells and their dye-swaps. Each dot or triangle represents one miRNA probe. Arrow indicates miR-103 probe, which is 44.26% of relative control intensity over the course of 24 h. (B, C) real-time Q-PCR analysis of mature miR-103 (B) and pri-miR-103 expression (C) during 24 h IGF-1 treatment in crypt cells. Data were normalized to snoRNA-202 levels for mature miR-103 and β-actin for pri-miR-103. Values are means ± SEM run in triplicates, letters indicate significant differences between time points.</p

Image_2_Systematic review and meta-analysis of type B aortic dissection involving the left subclavian artery with a Castor stent graft.tif

ObjectiveDespite the rapid development of thoracic endovascular aortic repair (TEVAR), it is still a challenge to maintain the blood flow of the branch arteries above the aortic arch in Stanford type B aortic dissection involving the left subclavian artery (LSA). The Castor stent graft is an integrated, customized, single-branch stent that enables reconstruction of the LSA. The purpose of this systematic review and meta-analysis was to assess the efficacy of the Castor stent graft for type B aortic dissection.Materials and methodsAn extensive electronic literature search (PROSPERO registration number: CRD42022322146) was undertaken to identify all articles published up to August 2022 that described thoracic aortic repair with branch stents in the treatment of type B aortic dissection involving the LSA. The quality of the included studies was analyzed using the MINORS criteria. The primary outcome measures were the technical success rate, early mortality rate, endoleak rate, and 1-year survival rate. The secondary outcome measures were the stroke rate, left upper extremity ischemia rate, and target vessel patency rate.ResultsEleven studies involving 415 patients were eligible for this meta-analysis. The LSA was successfully preserved in all procedures. The technical success rate was 97.5% (95% CI: 0.953–0.991); the intraoperative endoleak rate was 0.1% (95% CI: 0.000–0.012); the intraoperative LSA patency rate was 99.52%; the intraoperative LSA stent deformation and stenosis rate was 0.15% (95% CI: 0.000–0.051); the early type I endoleak rate was 1.6% (95% CI: 0.003–0.035); the 30-day mortality rate was 0.96%; the early reintervention rate was 0.9% (95% CI: 0.000–0.040); and the perioperative stroke rate was 0% (95% CI: 0.000–0.005). The 1-year survival rate was 99.7% (95% CI: 0.976–1.000). The half-year LSA patency rate was 99.3%, the 1-year LSA patency rate was 97.58%, and the 2-year LSA patency rate was 95.23%. During the follow-up period, the leakage rate was 0.3% (95% CI: 0.000–0.017), the incidence of left upper extremity ischemia rate was 0.5% (95% CI: 0.000–0.035), and the deformation and stenosis rate of the LSA stent was 2.2% (95% CI: 0.06–0.046).ConclusionThis meta-analysis shows that endovascular repair of type B aortic dissection using the Castor stent-graft may be technically feasible and effective. However, this conclusion needs to be interpreted with caution, as the quality of evidence for all outcomes is between low and very low.Systematic review registration[https://www.crd.york.ac.uk/prospero/], identifier [CRD42022322146].</p

Data_Sheet_1_Systematic review and meta-analysis of type B aortic dissection involving the left subclavian artery with a Castor stent graft.docx

ObjectiveDespite the rapid development of thoracic endovascular aortic repair (TEVAR), it is still a challenge to maintain the blood flow of the branch arteries above the aortic arch in Stanford type B aortic dissection involving the left subclavian artery (LSA). The Castor stent graft is an integrated, customized, single-branch stent that enables reconstruction of the LSA. The purpose of this systematic review and meta-analysis was to assess the efficacy of the Castor stent graft for type B aortic dissection.Materials and methodsAn extensive electronic literature search (PROSPERO registration number: CRD42022322146) was undertaken to identify all articles published up to August 2022 that described thoracic aortic repair with branch stents in the treatment of type B aortic dissection involving the LSA. The quality of the included studies was analyzed using the MINORS criteria. The primary outcome measures were the technical success rate, early mortality rate, endoleak rate, and 1-year survival rate. The secondary outcome measures were the stroke rate, left upper extremity ischemia rate, and target vessel patency rate.ResultsEleven studies involving 415 patients were eligible for this meta-analysis. The LSA was successfully preserved in all procedures. The technical success rate was 97.5% (95% CI: 0.953–0.991); the intraoperative endoleak rate was 0.1% (95% CI: 0.000–0.012); the intraoperative LSA patency rate was 99.52%; the intraoperative LSA stent deformation and stenosis rate was 0.15% (95% CI: 0.000–0.051); the early type I endoleak rate was 1.6% (95% CI: 0.003–0.035); the 30-day mortality rate was 0.96%; the early reintervention rate was 0.9% (95% CI: 0.000–0.040); and the perioperative stroke rate was 0% (95% CI: 0.000–0.005). The 1-year survival rate was 99.7% (95% CI: 0.976–1.000). The half-year LSA patency rate was 99.3%, the 1-year LSA patency rate was 97.58%, and the 2-year LSA patency rate was 95.23%. During the follow-up period, the leakage rate was 0.3% (95% CI: 0.000–0.017), the incidence of left upper extremity ischemia rate was 0.5% (95% CI: 0.000–0.035), and the deformation and stenosis rate of the LSA stent was 2.2% (95% CI: 0.06–0.046).ConclusionThis meta-analysis shows that endovascular repair of type B aortic dissection using the Castor stent-graft may be technically feasible and effective. However, this conclusion needs to be interpreted with caution, as the quality of evidence for all outcomes is between low and very low.Systematic review registration[https://www.crd.york.ac.uk/prospero/], identifier [CRD42022322146].</p

Image_1_Systematic review and meta-analysis of type B aortic dissection involving the left subclavian artery with a Castor stent graft.tif

ObjectiveDespite the rapid development of thoracic endovascular aortic repair (TEVAR), it is still a challenge to maintain the blood flow of the branch arteries above the aortic arch in Stanford type B aortic dissection involving the left subclavian artery (LSA). The Castor stent graft is an integrated, customized, single-branch stent that enables reconstruction of the LSA. The purpose of this systematic review and meta-analysis was to assess the efficacy of the Castor stent graft for type B aortic dissection.Materials and methodsAn extensive electronic literature search (PROSPERO registration number: CRD42022322146) was undertaken to identify all articles published up to August 2022 that described thoracic aortic repair with branch stents in the treatment of type B aortic dissection involving the LSA. The quality of the included studies was analyzed using the MINORS criteria. The primary outcome measures were the technical success rate, early mortality rate, endoleak rate, and 1-year survival rate. The secondary outcome measures were the stroke rate, left upper extremity ischemia rate, and target vessel patency rate.ResultsEleven studies involving 415 patients were eligible for this meta-analysis. The LSA was successfully preserved in all procedures. The technical success rate was 97.5% (95% CI: 0.953–0.991); the intraoperative endoleak rate was 0.1% (95% CI: 0.000–0.012); the intraoperative LSA patency rate was 99.52%; the intraoperative LSA stent deformation and stenosis rate was 0.15% (95% CI: 0.000–0.051); the early type I endoleak rate was 1.6% (95% CI: 0.003–0.035); the 30-day mortality rate was 0.96%; the early reintervention rate was 0.9% (95% CI: 0.000–0.040); and the perioperative stroke rate was 0% (95% CI: 0.000–0.005). The 1-year survival rate was 99.7% (95% CI: 0.976–1.000). The half-year LSA patency rate was 99.3%, the 1-year LSA patency rate was 97.58%, and the 2-year LSA patency rate was 95.23%. During the follow-up period, the leakage rate was 0.3% (95% CI: 0.000–0.017), the incidence of left upper extremity ischemia rate was 0.5% (95% CI: 0.000–0.035), and the deformation and stenosis rate of the LSA stent was 2.2% (95% CI: 0.06–0.046).ConclusionThis meta-analysis shows that endovascular repair of type B aortic dissection using the Castor stent-graft may be technically feasible and effective. However, this conclusion needs to be interpreted with caution, as the quality of evidence for all outcomes is between low and very low.Systematic review registration[https://www.crd.york.ac.uk/prospero/], identifier [CRD42022322146].</p