Phenotypic modulation of VSMCs by AG490 after AA procedure.

<p>The mRNA levels of ASMA, SM2, and SM22α was lowered after AA, while that of osteopoitin and SMemb were raised markedly and peaked at 7^th day after AA. At the 14^th day time point, expression of SM2 and SM22α began to increase but still lower than Sham group, and expression of osteopoitin and SMemb decreased to lower levels. AG490 down-regulated the osteopoitin and SMemb expression, and had a trend to increase the ASMA and SM2 mRNA levels. All the results were normalized to Sham. n = 4 anastomoses. (^* p<0.05, ^** p<0.01 versus AA+AG490 respectively).</p

activation features of JAK2/STAT3 in the vessel wall after AA.

<p><b>A–F</b>: p-STAT3 positive cells (red) in Sham group and AA group determined by immunofluorecence (Samples: 7 days after AA; Arrows indicated the typical positive cells. n = 3 anastomoses. Bar = 100 µm). <b>G–I</b>: Representative bands of target proteins in Sham group and AA groups at 3, 7, and 14 days after surgery (n = 4 anastomoses). The analysis showed the expression p-JAK2 and p-STAT3 increased and peaked at 7 days after the AA procedure (^** p<0.01 versus Sham group).</p

AG490 inhibited the up-regulation of p-JAK2, p-STAT3 and PCNA, and attenuated cell proliferation after AA.

<p><b>A</b>: AG490 down-regulated the activation of JAK2/STAT3 in the AA+AG490 group on the 7^th day (n = 4 anastomoses). <b>B–D</b>: mRNA levels of PCNA were up-regulated after AA, and inhibited markedly after treating with AG490 on the 7^th day. Protein levels of PCNA were increased and peaked at the 7^th day after AA, and down-regulated in AA+AG490 group (n = 4 anastomoses). <b>E–F:</b> Morphometry analysis of CCA on the 7^th and 14^th day. Quantification analysis results were illustrated by the outside radius (R_o)/inside radius (R_i) ratio. Bar = 500 µm. n = 3 anastomoses. (^* p<0.05 versus AA+AG490 group (7^th day); ^** p<0.01 versus Sham group; ## p<0.01, §§ p<0.01; NS: no significant difference).</p

Re-endothelialization assessment on AA site at different experimental time points.

<p>Quantification analysis of the blue staining area demonstrated heavy staining in and around the AA sites, which peaked at the 3^rd day, and regressed to normal level at the 30^th day after AA. No statistic difference was found between the AA+DMSO and AA+AG490 groups at all indicated time points. n = 3 anastomoses.</p

sj-pdf-1-jcb-10.1177_0271678X231205078 - Supplemental material for Growth arrest specific protein 6 alleviated white matter injury after experimental ischemic stroke

Supplemental material, sj-pdf-1-jcb-10.1177_0271678X231205078 for Growth arrest specific protein 6 alleviated white matter injury after experimental ischemic stroke by Junqiu Jia, Siyi Xu, Jinglong Hu, Yonghui Gan, Min Sun, Shengnan Xia, Xinyu Bao, Meijuan Zhang and Yun Xu in Journal of Cerebral Blood Flow & Metabolism</p

Human Urinary Kallidinogenase Promotes Angiogenesis and Cerebral Perfusion in Experimental Stroke

<div><p>Angiogenesisis a key restorative mechanism in response to ischemia, and pro-angiogenic therapy could be beneficial in stroke. Accumulating experimental and clinical evidence suggest that human urinary kallidinogenase (HUK) improves stroke outcome, but the underlying mechanisms are not clear. The aim of current study was to verify roles of HUK in post-ischemic angiogenesis and identify relevant mediators. In rat middle cerebral artery occlusion (MCAO) model, we confirmed that HUK treatment could improve stroke outcome, indicated by reduced infarct size and improved neurological function. Notably, the ¹⁸F-FDG micro-PET scan indicated that HUK enhanced cerebral perfusion in rats after MCAO treatment. In addition, HUK promotespost-ischemic angiogenesis, with increased vessel density as well as up-regulated VEGF andapelin/APJ expression in HUK-treated MCAO mice. In endothelial cell cultures, induction of VEGF and apelin/APJ expression, and ERK1/2 phosphorylation by HUK was further confirmed. These changes were abrogated by U0126, a selective ERK1/2 inhibitor. Moreover, F13A, a competitive antagonist of APJ receptor, significantly suppressed HUK-induced VEGF expression. Furthermore, angiogenic functions of HUK were inhibited in the presence of selective bradykinin B1 or B2 receptor antagonist both <i>in vitro</i> and <i>in vivo</i>. Our findings indicate that HUK treatment promotes post-ischemic angiogenesis and cerebral perfusion via activation of bradykinin B1 and B2 receptors, which is potentially due to enhancement expression of VEGF and apelin/APJ in ERK1/2 dependent way.</p></div

Additional file 1 of A novel starch-active lytic polysaccharide monooxygenase discovered with bioinformatics screening and its application in textile desizing

Additional file 1: Table S1. Physicochemical properties of 34 members in AA13 family. Table S2. Ten sequences with the highest scores obtained by HMM scanning of the Aspergillaceae database. Figure S1. Multiple sequence alignment analysis of 34 members in AA13 family. The red box highlighted the three conserved histidines and the double N/Q/E-x-F/Y-like motif. Figure S2. Annotations of domains for candidate expression sequences

HUK enhanced cerebral blood perfusion and promoted angiogenesis in stroke rats.

<p>(A) Representative images of ¹⁸F-FDG PET/CT in HUK group and vehicle group at 1 d and 14 d after stroke. (B) Uptake of ¹⁸F-FDG in HUK or vehicle-treated rats at different time points after stroke. *<i>p</i><0.05 <i>versus</i> vehicle group; **<i>p</i><0.01 <i>versus</i> vehicle group, N = 10 per group. (C) Left panel showed the typical detected areas of CD31 staining in rats brain indicated in boxed area. Right panel demonstrated representativeimmunofluorescent images stained by CD31 (green) 7 days after reperfusion. Scale bar = 100 μm. (D) CD31 positive vessels were counted and analyzed at indicated time point. Data was presented as capillary density reflected by number of CD31 positive vessels per mm². #<i>p</i><0.05 <i>versus</i> sham group; ##<i>p</i><0.01 <i>versus</i> sham group; *<i>p</i><0.05 <i>versus</i> vehicle group; **<i>p</i><0.01 <i>versus</i> vehicle group. N = 6 per group. (E) Quantitative data of CD31 mRNA levels at 1, 3, 7, and 14 d after stroke onset. # <i>p</i><0.05 <i>versus</i> sham group; *<i>p</i><0.05 <i>versus</i> control group. N = 6 per group.</p

HUK enhanced VEGF, Apelin/APJ expression in stroke rat brain.

<p>MCAO rats with or without HUK treatment were sacrificed and mRNA was extracted for the detection of apelin (A), APJ (B), VEGF (C) levels by real-time PCR. N = 6 per group. #<i>p</i><0.05 <i>versus</i> sham group; ##<i>p</i><0.01 <i>versus</i> sham group; *<i>p</i><0.05 <i>versus</i> control group; **<i>p</i><0.01 <i>versus</i> control group. Protein level of apelin in rat brain was determined by ELISA (F), while protein level of VEGF in rat brain was measured by Western blot (D). Quantitative data of VEGF blots at indicated time points (F). #<i>p</i><0.05 <i>versus</i> sham group; *<i>p</i><0.05 <i>versus</i> vehicle group, N = 6 repeats.</p

HUK enhanced VEGF, Apelin/APJ expression in stroke rat brain.

<p>MCAO rats with or without HUK treatment were sacrificed and mRNA was extracted for the detection of apelin (A), APJ (B), VEGF (C) levels by real-time PCR. N = 6 per group. #<i>p</i><0.05 <i>versus</i> sham group; ##<i>p</i><0.01 <i>versus</i> sham group; *<i>p</i><0.05 <i>versus</i> control group; **<i>p</i><0.01 <i>versus</i> control group. Protein level of apelin in rat brain was determined by ELISA (F), while protein level of VEGF in rat brain was measured by Western blot (D). Quantitative data of VEGF blots at indicated time points (F). #<i>p</i><0.05 <i>versus</i> sham group; *<i>p</i><0.05 <i>versus</i> vehicle group, N = 6 repeats.</p