Objective:To investigate the predictive value of YKL-40 at admission on stroke-associated pneumonia (SAP) and poor outcome in patients with acute ischemic stroke (AIS).Methods:Patients with AIS admitted to Taixing People’s Hospital from February 2020 to March 2021 were enrolled prospectively. The poor outcome was defined as 3-6 points on the modified Rankin Scale at 90 d after onset. Multivariate logistic regression analysis was used to determine the independent predictors of SAP and poor outcome, and the predictive value of serum YKL-40 on SAP and poor outcome was evaluated by receiver operating characteristic (ROC) curve. Results:A total of 377 patients with AIS were enrolled. The median serum YKL-40 was 127.16 μg/L. One hundred and four patients (27.6%) had SAP, and 126 (33.4%) had poor outcomes at 90 d after onset. Multivariate logistic regression analysis showed that after adjusting for confounding factors, YKL-40 was the independent predictors of SAP (odds ratio [ OR] 1.005, 95% confidence interval [ CI] 1.003-1.008; P=0.001) and poor outcome at 90 d ( OR 1.009, 95% CI 1.006-1.011; P=0.001). The ROC curve analysis showed that the area under the curve of YKL-40 for predicting SAP was 0.769 (95% CI 0.713-0.824; P<0.001), the best cutoff value was 168.70 μg/L, and the sensitivity and specificity were 71.2% and 75.1% respectively; the area under the curve of YKL-40 for predicting poor outcome at 90 d was 0.787 (95% CI 0.735-0.840; P<0.001), the best cutoff value was 195.56 μg/L, and the sensitivity and specificity were 68.3% and 84.1% respectively. Conclusion:Higher serum YKL-40 at admission has a good predictive value for SAP and poor outcome at 90 d in patients with AIS.

Objective:To investigate the predictive value of serum hypersensitive C-reactive protein (hs-CRP) for stroke-associated pneumonia (SAP) in patients with acute ischemic stroke (AIS) who received intravenous thrombolysis.Methods:From May 2015 to April 2017, the clinical data of the patients with AIS treated with intravenous thrombolysis in Nanjing First Hospital were collected retrospectively. Multivariate logistic regression analysis was used to determine the independent risk factors for SAP in patients with AIS after intravenous thrombolysis. Receiver operating characteristic (ROC) curve and nomogram-based methods were used to analyze the predictive value of hs-CRP for SAP. Results:A total of 243 patients with AIS who received intravenous thrombolysis were included, and 63 (34.6%) of them had SAP. There were significant differences in age ( P=0.006), leukocyte count ( P=0.044), fasting blood glucose level ( P=0.003), serum hs-CRP level ( P=0.001), hs-CRP classification ( P=0.001) and dysphagia rate ( P=0.035) between the SAP group and non-SAP group. Multivariate logistic regression analysis showed that after adjusting for the confounding factors, taking the first quartile of serum hs-CRP level as a reference, the third quantile (odds ratio [ OR] 18.790, 95% confidence interval [ CI] 4.771-74.007; P=0.001) and the fourth quantile ( OR 54.054, 95% CI 12.248-324.088; P=0.001) of hs-CRP were the independent predictors of SAP. The area under the ROC curve of the baseline serum hs-CRP level for predicting SAP was 0.805 (95% CI 0.742-0.868; P<0.001). When the optimal cut-off value of hs-CRP was 5.54 mg/L, the sensitivity and specificity of predicting SAP were 76.11% and 76.19%, respectively. The analysis of nomogram also showed that hs-CRP was an independent predictor of SAP (consistency index 0.862, 95% CI 0.738-0.986; P<0.001). Conclusions:The increased serum hs-CRP was an independent predictor of SAP in patients with AIS receiving intravenous thrombolysis, and had a higher predictive value.

Objective Based on the gene expression omnibus(GEO)database,bioinformatics methods were employed to analyze the expression characteristics of hypoxia-related differentially expressed genes(HRDEGs)in ischemic stroke,and key genes were screened,to provide important support for a deeper understanding of ischemic stroke.Methods The GSE16561 and GSE58294 datasets were downloaded from the GEO database,and Python software was used for data integration.The Combat method was employed to eliminate batch effects while retaining disease grouping characteristics.Principal component analysis was conducted to reduce dimensionality of the data before and after batch effect removal,and intraclass correlation coefficient(ICC)testing was performed on the ischemic stroke and normal control groups.Gene set enrichment analysis(GSEA)and single-sample GSEA were conducted on the merged and batch effects eliminated dataset,with a nominal P-value(NOM P-val)＜0.05 and false discovery rate P-value(FDR P-val)＜0.25 used as criteria to select significantly different gene sets.Differential expression genes between the ischemic stroke samples and normal control samples after merging and eliminating batch effects of the GSE16561 and GSE58294 datasets were identified using R software,with an absolute value of log2 gene expression fold change(FC)≥0.58 and adjusted P-value(Padj)＜0.05 as selection criteria.Intersection with hypoxia-related genes obtained from the National Center for Biotechnology Information(NCBI)in the United States yielded the HRDEGs.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were performed on the HRDEGs,and the STRING database was used to construct a protein-protein interaction network of differentially expressed genes.The top 10 key genes were filtered using Cytoscape 3.8 software.Results The ICC analysis results showed excellent consistency in the ischemic stroke and normal control samples after batch effect removal,with ICC values of 0.94 and 0.98 for the GSE16561 and GSE58294datasets,respectively.GSEA results demonstrated significant enrichment of 34 gene sets in the stroke samples in the newly merged and batch effects removed dataset from GSE16561 and GSE58294,leading to the identification of 404 differentially expressed genes(all with Padj＜0.05),including 354 upregulated genes and 50 downregulated genes.Intersection with hypoxia-related genes yielded 64 HRDEGs.GO enrichment analysis indicated significant enrichment of HRDEGs in vesicle lumen,cytoplasmic vesicle lumen,secretory granule lumen,with molecular functions such as amide binding,peptide binding,phospholipid binding,and enzyme inhibitor activity.These genes are primarily involved in the positive regulation of cytokine production,regulation of immune response,response to bacterium-derived molecules,and response to lipopolysaccharide,among other biological processes.KEGG enrichment analysis revealed enrichment of HRDEGs in pathways related to lipid and atherosclerosis,Salmonella infection,neutrophil extracellular trap formation,nucleotide-binding oligomerization domain-like receptor signaling pathway,protein glycosylation in cancer,tuberculosis,and necroptosis.Based on the protein-protein interaction network,10 key genes were identified,including arginase1(ARG1),caspase1(CASP1),interleukin1 receptor type 1(IL-1R1),integrin subunit alpha M(ITGAM),matrix metalloproteinase9(MMP9),prostaglandin-endoperoxide synthase 2(PTGS2),signal transducer and activator of transcription 3(STAT3),Toll-like receptor2(TLR2),TLR4,and TLR8.Conclusion This study has identified 10 key genes associated with ischemic stroke and hypoxia through bioinformatics mining,which maybe provid potential targets for subsequent research and diagnostic and therapeutic interventions.

Objective To investigate the pathological damage to and inflammation of different intestinal segments in a rat model of severe hemorrhage,and to explore the effect of polarization of intestinal macrophage on the pathophysiology of intestinal inflammation.Methods Male Wistar rats were randomly divided into two groups:the sham operation group and hemorrhage group.In the hemorrhage group,40％of the total blood volume was lost in 25-30 minutes,while in the sham operation group,only the femoral artery and vein were intubated without bleeding.The rats were killed at 0,3,6,12 and 24 hours.The entire intestine was isolated quickly,and sections of the intestine were cut at the duodenum,jejunum,ileocecal junction,colon and rectum for histopathological evaluation.ELISA was adopted to determine related inflammation factors while multi-color immunohistochemistry was used to calculate macrophage surface markers.The data was statistically analyzed.Results(1)Compared with the sham group,there was no significant difference in colon histology at 3 h and 6 h,but significant difference was detected in rectum scores only at 24 h.The scores of other intestinal segments were significantly different at each time point.The severity of ileocecal and colonic lesions after bleeding increased with time.The duodenum,jejunum and ileocecum were more critically injured at 3 h than the rectum at 6 h.The injury to the duodenum,jejunum,ileum and colon was much more pronounced than to the rectum at 12 h.(2)The expressions of TNF-α and IL-1β in the rectum were increased significantly at 12 h post operation.The expressions of IL-1β,TNF-α in the jejunum increased obviously at 3 h and 6 h,respectively.(3)Three hours after severe bleeding,the level of macrophages in the jejunum and ileocececal area increased significantly,and the percentage of M1 macrophages was higher.After 6 hours,the proportion of M2 macrophages in the jejunum and M1 macrophages decreased significantly.After 3 hours,the percentage of M1 macrophages in the colon decreased,but that of M2 macrophages increased.The proportion of M2 polarized macrophages in the duodenum and rectum increased at 3 h after severe bleeding but decreased at 6 h.Conclusion Pathological damage to intestinal sections after bleeding varies depending on the time,and is correlated with the inflammatory level of macrophages.

OBJECTIVE To investigate the distribution and regulation of G-quadruplex(G4)in enzymes related to glycolysis.METHODS The sequences of the transcription start site(TSS)region upstream of 1500 bp to the 5'-Untranslated region in 200 enzymes and their subtypes in glycolysis were selected for bioinformatics analysis.Related enzymes in glycolysis containing putative G-quadru-plex-forming sequences(PQSs)were identified.Circular Dichroism and Native polyacrylamide gel elec-trophoresis were used to verify the formation of G4.The ExonucleaseⅠhydrolysis assay was used to validate the stability of the formed G4 under 0,0.5,2,8,16,and 32 min.A reporter gene plasmid was constructed by inserting specific fragments of the related enzymes before the luciferase expression sequence.The dual-luciferase reporter assay system validate the expression level of luciferase to assess the impact of G4 on promoter activity.Real-time quantitative PCR was performed to validate the transcriptional regulatory role of G4 by detecting the mRNA levels of luciferase.RESULTS ①Bioin-formatics analysis showed that out of the 200 glycolysis-related enzymes,12 contained PQSs.Based on the analysis of the length and structure of PQSs,aldolase A(ALDOA)and phosphoglycerate mutase 2(PGAM2)proved to be able to form stable G4.② ALDOA and PGAM2 had the maximum positive absorption peak at 260 nm and maximum negative absorption peak at 240 nm.Both of them could form a G4 at the same time.③After digestion with ExonucleaseⅠ,ALDOA and PGAM2 showed no significant hydrolysis and demonstrated the stability of G4 structures.However,both of them could be gradually hydrolyzed after mutations in their PQSs.④ After PQS mutation of ALDOA and PGAM2,the mRNA levels and expression of downstream luciferase of ALDOA were significantly increased(P<0.05,P<0.01),while PGAM2 was significantly decreased(P<0.05,P<0.01).CONCLUSION The gene sequences of glycolysis-related enzymes and their subtypes contain a large number of PQSs.ALDOA and PGAM2 can form stable G4 and perform transcriptional regulatory functions.

In order to improve the wearing comfort and bearing effectiveness of the exoskeleton, based on the prototype and working mechanism analysis of a relaxation wearable system for knee exoskeleton robot, the static optimization synthesis and its method are studied. Firstly, based on the construction of the virtual prototype model of the system, a comprehensive wearable comfort evaluation index considering the factors such as stress, deformation and the proportion of stress nodes was constructed. Secondly, based on the static simulation and evaluation index of system virtual prototype, multi-objective genetic optimization and local optimization synthesis of armor layer topology were carried out. Finally, the model reconstruction simulation data confirmed that the system had good wearing comfort. Our study provides a theoretical basis for the bearing performance and prototype construction of the subsequent wearable system.
Humans ; Exoskeleton Device ; Computer Simulation ; Emotions ; Knee Joint