Objective:To screen the differentially expressed immune-related genes(DEIRGs)in in-stent restenosis(ISR),and to analyze the immune cell infiltration in ISR,and to clarify the mechanism of occurrence and development of ISR.Methods:The mRNA gene expression data of GSE46560 dataset samples were downloaded from the Gene Expression Omnibus(GEO),and divided into ISR group and non-ISR group.The"Limma"package in R software was used to identify the differentially expressed genes(DEGs)which were then intersected with immune-related genes(IRGs)to identify the DEIRGs in ISR;R software was used for Gene Ontology(GO)functional enrichment andalysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis on DEIRGs;the STRING database was used to construct the protein-protein interaction(PPI)network,which was visualized and analyzed for Hub genes by Cytoscape software;the receiver operating characteristic(ROC)curve of the Hub genes were plotted,and the area under the curve(AUC)was calculated and the diagnostic value was evaluated;CIBERSORT software was used to analyze the immune cell infiltration in ISR;Pearson correlation analysis was used to analyze the relationships between the immune cells and the relationships between the immune cells and key genes.Results:A total of 331 DEGs were identified(P<0.05,|log2FC|>1),including 176 upregulated genes and 155 downregulated genes,and 38 DEIRGs were obstained.The GO functional enrichment analysis results showed that the DEIRGs were mainly enriched in biological processes(BP)such as defense response,immune response,and immune system;in cellular components(CC),the DEIRGs were located primarily in the extracellular region and cytoplasmic membrane;and in molecular functions(MF),the DEIRGs were mainly involved in regulating signaling receptor binding and cytokine receptor activity.The KEGG signaling pathway enrichment analysis results indicated that the DEIRGs in ISR were primarily enriched in the phosphatidylinositol 3-kinase/protein kinase B(PI3K-AKT)and transforming growth factor-β(TGF-β)signaling pathways.In the PPI network,CD19 had the highest node among the top 10 Hub genes.Compared with non-ISR group,the expression level of the CD19 gene in the samples in ISR group was increased(P<0.05).The AUC value in the ROC curve of CD19 gene expression was 0.92(P<0.05).The immune cell infiltration analysis results showed that compared with non-ISR group,the infiltration level of T lymphocyte follicular helper(Tfh)cells in the patients in ISR group were increased(P<0.05),the infiltration levels of immature B lymphocytes,CD8+T lymphocytes,naive CD4+T lymphocytes,and M0 macrophages were increased,but the differences were not statistically significant(P>0.05),while the infiltration levels of memory B lymphocytes,activated memory CD4+T lymphocytes,regulatory T cells,resting natural killer(NK)cells,activated NK cells,monocytes,resting mast cells,and neutrophils were decreased,but the differences were not statistically significant(P>0.05).There were positive correlations between Tfh cells and M0 macrophages and resting mast cells(r=0.88,P<0.05;r=0.68,P<0.05),and there were negative correlations between Tfh cells and monocytes and neutrophils(r=-0.49,P<0.05;r=-0.42,P<0.05).Conclusion:CD19 may influence the occurrence and development of ISR by regulating the activation of the PI3K-AKT signaling pathway to affect the Tfh and B lymphocytes.CD19 can serve as a biomarker for the diagnosis of ISR.

Objective:To investigate key differential expressed genes(DEGs),enriched biological functions and signaling pathways in exosomes derived from angiosarcoma cells ISOHAS and angiosarcoma stem-like cells Sphere by bioinformatics analysis,to provide new therapeutic targets for angiosarcoma.Methods:Transcriptome sequencing of exosomes derived from ISOHAS and Sphere were performed to screen for DEGs by|log2FC|>2 and FDR<0.05 as criteria.Bioinformatics analysis was used to enrich GO and KEGG pathways of DEGs to identify biological functions and signaling pathways of enriched DEGs.STRING database was used to screen key DEGs,and iPath was used to visualize to identify metabolic pathways enriched by DEGs.Results:Transcriptome sequencing results showed that 91 DEGs were identified in exosomes derived from ISOHAS and Sphere.GO and KEGG pathway enrichment analysis demonstrated that main biological functions and signaling pathways enriched by DEGs were response to glucocorticoid and TNF signaling pathway,respectively.STRING database demonstrated that TNF and IL-6 were key DEGs.iPath metabolic network analysis demonstrated that DEGs were mainly identified in lipid metabolism and nucleotide metabolism.Conclusion:Sphere-derived exosomes may influence occurrence and development of angiosarcoma by carrying key genes TNF and IL-6 to interfere with glucocorticoid response,TNF signaling pathway,lipid metabolism,nucleotide metabolism and other biological functions and signaling pathways,providing new ideas for therapeutic targets for angiosarcoma.

Objective:To observe any effect of long-distance application of Oral Reading for Language with Aphasia (ORLA) training on patients with post-stroke aphasia.Methods:A total of 42 stroke survivors with aphasia were randomly divided into an inpatient group, and two online groups, each of 14. All three groups had their routine rehabilitation treatment supplemented with ordinary multimodal language therapy and ORLA. The inpatient group completed the routine in the rehabilitation treatment room, while the online groups completed it at home using Tencent video conferencing software. The conventional multimodal language therapy was conducted once daily, 5 days a week for 4 weeks. For the inpatient group and online group 2 the daily session lasted 30 minutes, while for the online 1 groups the daily length was doubled. The ORLA therapy was also conducted once daily, 5 days a week for 4 weeks, for the inpatient group and online group 2 the daily session lasted 1h, while for the online 1 groups the daily length was 30min.The speech function, reading ability and life quality of the three groups were evaluated before and after the intervention using the Western Aphasia Battery, the Chinese Standard Aphasia Examination Scale and the Chinese version of the Stroke Aphasia Quality of Life Scale.Results:After treatment, the average aphasia quotient (AQ), reading, naming and quality of life scores in all three groups had improved significantly compared with those before treatment. And related language ability scores (such as retelling, fluency, information volume, listening comprehension, etc.) had also improved significantly in all three groups. However, the average AQ, reading, and oral fluency scores of the inpatient group and online group 2 were significantly higher than those of online group 1. Significant improvement was also observed in the reading aloud and life quality of all three groups, but the average improvement in reading aloud was significantly greater in the inpatient group and in online group 2 compared to online group 1. The average life quality of the online groups was significantly superior to that of the inpatient group.Conclusion:Medium- and high-intensity ORLA synchronous remote speech rehabilitation can significantly improve the speech ability, reading ability and life quality of aphasic stroke survivors.

Objective:To observe any effect of combining vibroacoustic therapy with audio-visual feedback on the swallowing and the emotional state of persons with a swallowing disorder after a brainstem stroke.Methods:Fifty brainstem stroke survivors with dysphagia were randomly divided into a control group and a combination group, each of 25. In addition to routine swallowing function training, the control group received additional direct feeding training, while the combination group was provided with vibroacoustic feeding training and swallowing stimulation from an animated audio-visual presentation. Before and after the treatment, the subjects′ swallowing function was quantified using the Standard Swallowing Assessment (SSA) scale and the Gugging swallowing screen (GUSS). The Positive and Negative Emotions Scale (PNAS) was used to quantify their emotional state.Results:After the treatment the average SSA scores of both groups had decreased significantly, while the average GUSS and PNAS scores had increased significantly. The average improvements in all three measures were significantly greater in the combination group than in the control group.Conclusion:In addition to routine swallowing function training, feeding training applying vibroacoustics and audio-visual stimulation can significantly improve the feeding function and emotional state of brainstem stroke survivors with dysphagia.

N6-Methyladenosine (m⁶A) is the most abundant internal modification in eukaryotic mRNA, playing critical role in various bioprocesses. Like other epigenetic modifications, m⁶A modification can be catalyzed by the methyltransferase complex and erased dynamically to maintain cells homeostasis. Up to now, only two m⁶A demethylases have been reported, fat mass and obesity-associated protein (FTO) and alkylation protein AlkB homolog 5 (ALKBH5), involving in a wide range of mRNA biological progress, including mRNA shearing, export, metabolism and stability. Furthermore, they participate in many significantly biological signaling pathway, and contribute to the progress and development of cancer along with other diseases. In this review, we focus on the studies about structure, inhibitors development and biological function of FTO and ALKBH5.

Objective:To observe any effect of dry needling of myofascial trigger points on substance P and synaptophysin expression in the spinal dorsal horn.Methods:Sixty-four Sprague-Dawley rats were randomly divided into a control group ( n=16) and a model group ( n=48). Myofascial trigger points were induced in the model group by a blunt strike and eccentric running. That group was then randomly divided into a no-treatment group ( n=15), a massage group ( n=16), and a dry needling group (16 rats). The rats in the two treatment groups received 4 weeks of dry needling or Chinese massage. Pressure pain thresholds were recorded before the experiment and after the 4 weeks. The content of substance P and synaptophysin in the spinal dorsal horn were measured using immunoblotting and immunohistochemistry. Results:After the treatment 14 rats (93%) in the model group had trigger points, significantly higher than the 8 rats (50%) in the massage group and the 7 rats (44%) in the dry needling group. After treatment, the average pressure pain thresholds of the no-treatment and massage groups was significantly lower than the control group′s average, while the difference between the dry needling group and the control group was not significant. The average pressure pain threshold had improved significantly in the no-treatment group, the massage group and the dry needling group, and the averages of the massage group and the dry needling group were significantly higher than that of the no-treatment group. The level of substance P was significantly higher in the no-treatment group than in the other three groups and the ratio of substance P to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH)was significantly higher. The substance P: GAPDH ratio of the massage group was significantly higher than that of the control group.Conclusions:Dry needling and massage are effective in relieving myofascial pain, at least in rats. Both can reduce the content of substance P in the spinal dorsal horn.

Objective : To explore the role and its clinical significance of P300 in biphasic differentiation and epithelial mesenchymal transition ( EMT) process in synovial sarcoma (SS) by detecting the expression of histone acetyltransferase P300 and EMT related molecules. Methods : 40 cases of SS paraffin embedded tissue samples were collected , and the SYT⁃SSX fusion gene subtype was detected by RT⁃PCR. Immunohistochemistry was used to detect the expression of P300 and EMT⁃related molecules. : Results Conclusion P300 may be involved in the biphasic differentiation and the EMT process of SS , suggesting that P300 plays a certain role in SS invasion and migration.

Objective To study the sensitivity and specificity of parameter Corvis biomechanical index (CBI)— a new biomechanical index of Corvis ST in the diagnosis of keratoconus and evaluate the role of CBI in the diagnosis of keratoconus and the change of biomechanic.Methods A diagnostic trial study was adopted,and 66 eyes from 49 keratoconic patients (keratoconus group) and 91 right eyes from 91 myopic patients (control group) from April in 2018 to August in 2018 in Henan Eye Hospital were enrolled.Pentacam and Corvis ST examinations were performed by the same operator after the basic eye examinations.Using the evaluation of diagnostic test,consistency test,and receiver operating characteristic (ROC) curve analysis,gain the outcome of the sensitivity,specificity,consistency,Youden index,and area under the ROC curve of the parameter CBI.This study protocol was approved by Ethic Committee of Henan Eye Hospital and followed the Declaration of Helsinki.Written informed consent was obtained from each patient prior to any medical examination.Results Sixty eyes were diagnosed as keratoconus by the parameter CBI of Corvis ST.Evaluation of diagnosis test:the sensitivity was 97.0％,and the specificity was 97.8％;consistency check:Kappa =0.948,P＜0.05.ROC curve analysis:the sensitivity was 98.5％,the specificity was 96.8％,Youden index was 96.3％,P＜0.000 1 and AUC was 0.996.Conclusions CBI can separate healthy eyes from keratoconic eyes with highly sensitivity and specificity,which was highly consistency with results of Rabinowitz keratoconus diagnostic criteria.CBI could be used as a new biomechanical indicator for the diagnosis of keratoconus.

Objective To observe the gastric changes in adult male Sprague-Dawly (SD) rats irradiated by the single large dose electron beam,providing animal experimental evidence for intraoperative radiotherapy for gastric cancer.Methods Thirty-eight SD rats were randomly divided into the control and experimental groups.The stomach of the rats in the experimental group were subject to single 6 MeV 20 Gy irridiation by using the patent technology of Accurate Irradiation Experiment Table for Small Animal Radiation.The general conditions,gastric injury and body weight change were observed at different days following irradiation.Results The most severe gastric damage of rats was observed on the 14th d after irradiation.The gastric injury was gradually repaired accompanied with glandular atrophy at 28 d postirradiation,and the gastric injury was manifested as cellulose fibrinous repair on the 56th d after irradiation.Within 1 week post-irradiation,weight loss was noted in the experimental group,which significantly differed from the rats in the control group (P＜0.05).During the 2nd week,the body weight was increased in the experimental group,significantly lower compared with the rats in the control group (P＜ 0.05).The body weight of rats did not significantly differ between two groups at 6 weeks after irradiation (P＞ 0.05).Conclusions The most severe gastric injury is observed at 2 weeks after the single-dose 6 MeV electron beam 20 Gy irradiation,whereas no gastric perforation occurs.The gastric injury can be restored to normal status within 8 weeks following irradiation.

Objective: To investigate the interaction of Ca²⁺ protein TRPC1 and STIM1 in extracellular Ca²⁺ -sensing receptor (CaR)-induced extracellular Ca²⁺ influx and the production of nitric oxide (NO). Methods: Human umbilical vein endothelial cells (HUVECs) were cultured and incubated with CaR agonist spermine (activating store-operates cation channels (SOC) and receptor-operated channels (ROC)), CaR negative allosteric modulator Calhex231 (blocking SOC, activating ROC) and ROC analogue TPA (activating ROC, blocking SOC), protein kinase C (PKC) inhibitor Ro31-8220, PKCs and PKCμ inhibitor Go6967(activate SOC, blocking ROC), respectively. The interaction of TRPC1 and STIM1 was determined using the immunofluorescence methods. The interaction between TRPC1 and STIM1 were examined by Co-immuno precipitation. The HUVECs were divided into: TRPC1 and STIM1 short hairpin RNA group (shTRPC1+ shSTIM1 group), vehicle-TRPC1+ vehicle-STIM1 group and control group. The cells were incubated with four different treatments under the action of above mentioned interventions, intracellular Ca²⁺ concentration ([Ca²⁺ ]_i) was detected using the fluorescence Ca²⁺ indicator Fura-2/AM, the production of NO was determined by DAF-FM. Results: (1) The expression of TRPC1 and STIM1 proteins levels in HUVECs: Under the confocal microscope, TRPC1 and STIM1 protein expression showed masculine gender, both located in cytoplasm in the normal control group. Post incubation with Calhex231+ TPA, Ro31-8220 and Go6967, TRPC1 and STIM1 positioned in cytoplasm was significantly reduced, and the combined TRPC1 and STIM1 was also significantly reduced. (2) The interaction of TRPC1 and STIM1 in HUVECs: The relative ratios of Calhex231+ TPA+ Spermine+ Ca²⁺ group, Ro31-8220+ Spermine+ Ca²⁺ group and Go6976+ Spermine+ Ca²⁺ group STIM1/TRPC1 and TRPC1/STIM1 were as follows: (25.98±2.17)% and (44.10±4.01)%, (20.85±1.01)% and (46.31±3.47)%, (23.88±2.05)% and (39.65±2.91)%, which were significantly lower than those in the control group (100.00±4.66)% and (100.00±6.40)% and in the Spermine+ Ca²⁺ group (106.04±2.45)% and (107.78±2.66)% (all P<0.05). (3) The influence of joint TRPC1 and STIM1 transfection to four different drugs treated HUVECs on [Ca²⁺ ]_i and NO generation: The changes of two excitation fluorescence intensity ratio and NO net fluorescence intensity values were consistent, [Ca²⁺ ]_i and NO net fluorescence intensity values were significantly lower in the experimental group than the control group and the vehicle group (all P<0.05), while which were similar between the vehicle group and control group (all P>0.05). Conclusions Our results indicate that TRPC1 and STIM1 jointly regulate CaR-mediated Ca²⁺ influx and nitric oxide generation in HUVECs in the form of binary complex.