Objective To investigate the risk factors and significance of catheter dislocationat the venipuncture site after the implantation of a totally implantable venous access port(TIVAP)in patients with breast cancer.Methods From January 2019 to September 2021,1003 patients who underwent vein approach transfusion port implantation were divided into the catheter dislocation group(7 cases)and the non-catheter dislocation group(996 cases).Risk factors for post operative recurrence of catheter dislocationwere analyzed through univariate analysis and logistic regression analysis.Results The results of the univariate analysis indicated that the incidence of catheter dislocation in the group with age ≥ 60 years,axillary vein approach,and left-side puncture was higher than that in the control group,and the difference was statistically significant(P＜0.05).The BMI of the dislocation group[(27.06±2.16)kg/m2]was higher than that of the non-dislocation group[(25.09±3.33)kg/m2],there was a statistically significant difference between the two groups(P=0.05).The multivariate Logistic regression analysis showed that the catheterization approach and puncture side had no significant effect on catheter dislocation(P＞0.05);high BMI and age ≥ 60 years were independent risk factors for catheter dislocation complications(P＜0.05).Conclusion Axillary vein approach transfusion port implantation is relatively safe and reliable.Age ≥ 60years old and high BMI are independent risk factors affecting the complication of catheter dislodgement.

OBJECTIVE To develop a highly sensitive and specific quantitative real-time poly-merase chain reaction(qPCR)method for detecting the VGM-R02b(a gene therapy drug for glutaric acidemia type Ⅰ)gene in cynomolgus monkeys and analyze the biological distribution of VGM-R02b.METHODS A standard curve was constructed using the VGM-R02b standard plasmid[an adeno-associ-ated virus serotype 9(AAV9)capsid]on a qPCR platform.The detection method was optimized and validated for key parameters,including the quantitative range,accuracy,precision,dilution linearity,selectivity,specificity,stability,and parallelism.The established method was used to determine the target gene of VGM-R02b in the blood,brain,stomach,heart,liver,spleen,lung,kidney,thymus,and duodenum of cynomolgus monkeys on day 29(D29)and D92 after a single,unilateral intraventricular injection of VGM-R02b.The biodistribution of VGM-R02b in cynomolgus monkeys was analyzed.RESULTS A qPCR method for quantifying the VGM-R02b target gene in cynomolgus monkeys was established and validated.The standard curve demonstrated a quantitative range of 5.00×109 to 5.00×10^1 copies·μg-1 DNA,with excellent precision,accuracy,dilution linearity,selectivity,and specificity.The target gene in tissues remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 9 d,-60 to-80℃ for 90 d and after five freeze-thaw cycles.Similarly,the target gene in whole blood remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 93 d,-60 to-80℃ for 93 d and after five freeze-thaw cycles.Extracted nucleic acid samples also showed stability after being stored at room temperature for 4.25 h,2-8℃ for 24.16 h,-60 to-80℃ for 109 d and after five freeze-thaw cycles.The method was also applied to evaluate the biological distribution of the target gene in cynomolgus monkeys.In the control group,the target gene was undetectable on D29 and D92 post-administration,but in the drug administration group,the target gene was distributed across the tissues,with higher concen-trations observed in the brain,liver,spleen,and spinal cord,and there were no significant differences in the target gene content across the tissues between D29 and D92.CONCLUSION The established qPCR method is robust,reliable and suitable for determination of VGM-R02b target gene in cynomolgus monkeys.

Objective To investigate the risk factors and significance of catheter dislocationat the venipuncture site after the implantation of a totally implantable venous access port(TIVAP)in patients with breast cancer.Methods From January 2019 to September 2021,1003 patients who underwent vein approach transfusion port implantation were divided into the catheter dislocation group(7 cases)and the non-catheter dislocation group(996 cases).Risk factors for post operative recurrence of catheter dislocationwere analyzed through univariate analysis and logistic regression analysis.Results The results of the univariate analysis indicated that the incidence of catheter dislocation in the group with age ≥ 60 years,axillary vein approach,and left-side puncture was higher than that in the control group,and the difference was statistically significant(P＜0.05).The BMI of the dislocation group[(27.06±2.16)kg/m2]was higher than that of the non-dislocation group[(25.09±3.33)kg/m2],there was a statistically significant difference between the two groups(P=0.05).The multivariate Logistic regression analysis showed that the catheterization approach and puncture side had no significant effect on catheter dislocation(P＞0.05);high BMI and age ≥ 60 years were independent risk factors for catheter dislocation complications(P＜0.05).Conclusion Axillary vein approach transfusion port implantation is relatively safe and reliable.Age ≥ 60years old and high BMI are independent risk factors affecting the complication of catheter dislodgement.

OBJECTIVE To develop a highly sensitive and specific quantitative real-time poly-merase chain reaction(qPCR)method for detecting the VGM-R02b(a gene therapy drug for glutaric acidemia type Ⅰ)gene in cynomolgus monkeys and analyze the biological distribution of VGM-R02b.METHODS A standard curve was constructed using the VGM-R02b standard plasmid[an adeno-associ-ated virus serotype 9(AAV9)capsid]on a qPCR platform.The detection method was optimized and validated for key parameters,including the quantitative range,accuracy,precision,dilution linearity,selectivity,specificity,stability,and parallelism.The established method was used to determine the target gene of VGM-R02b in the blood,brain,stomach,heart,liver,spleen,lung,kidney,thymus,and duodenum of cynomolgus monkeys on day 29(D29)and D92 after a single,unilateral intraventricular injection of VGM-R02b.The biodistribution of VGM-R02b in cynomolgus monkeys was analyzed.RESULTS A qPCR method for quantifying the VGM-R02b target gene in cynomolgus monkeys was established and validated.The standard curve demonstrated a quantitative range of 5.00×109 to 5.00×10^1 copies·μg-1 DNA,with excellent precision,accuracy,dilution linearity,selectivity,and specificity.The target gene in tissues remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 9 d,-60 to-80℃ for 90 d and after five freeze-thaw cycles.Similarly,the target gene in whole blood remained stable after being stored at room temperature for 4 h,-15 to-25℃ for 93 d,-60 to-80℃ for 93 d and after five freeze-thaw cycles.Extracted nucleic acid samples also showed stability after being stored at room temperature for 4.25 h,2-8℃ for 24.16 h,-60 to-80℃ for 109 d and after five freeze-thaw cycles.The method was also applied to evaluate the biological distribution of the target gene in cynomolgus monkeys.In the control group,the target gene was undetectable on D29 and D92 post-administration,but in the drug administration group,the target gene was distributed across the tissues,with higher concen-trations observed in the brain,liver,spleen,and spinal cord,and there were no significant differences in the target gene content across the tissues between D29 and D92.CONCLUSION The established qPCR method is robust,reliable and suitable for determination of VGM-R02b target gene in cynomolgus monkeys.

Objective:To analyze gene expression data of esophageal adenocarcinoma(EAC)in the Gene Expression Omnibus(GEO)and The Cancer Genome Atlas(TCGA)databases and clarify the relationship between the potential core genes and tumor lymphocyte infiltration in the EAC,and to provide the molecular targets for the diagnosis and treatment of EAC.Methods:The high-throughput chip datasets GSE13898,GSE26886,GSE74553,and GSE92396,including EAC and normal esophageal tissues,were downloaded from the GEO database by searching for"esophageal adenocarcinoma".The limma package of R software was used to screen the differentially expressed genes(DEGs)in EAC tissue and esophageal normal tissue,and the common DEGs were obtained through Venn diagram.After the DEGs were analyzed by STRING database,the results were imported into Cytoscape software to screen the core genes and construct the protein-protein interaction(PPI)network.The Gene Expression Profiling Interactive Analysis(GEPIA)database was used to verify the expression levels of core genes.The UALCAN and Kaplan-Meier Plotter databases were used to analyze the correlations between the core genes and prognosis and clinical data of the EAC patients.The Tumor Immune Estimation Resource(TIMER)database was used to analyze the relationship between core genes and tumor immune infiltration.Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway enrichment analysis were performed to analyze the positively correlated genes of core genes obtained from the LinkedOmics database.Results:A total of 340 DEGs were obtained from the intersection of DEGs from the four GEO datasets,including 127 upregulated genes and 213 downregulated genes.After screening with the STRING database and Cytoscape software,the key core genes with the highest scores were secreted phosphoprotein 1(SPP1)and transforming growth factor beta 1(TGFB1).The GEPIA database analysis results showed that compared with esophageal normal tissue,the expression levels of SPP1 and TGFB1 mRNA in cancer tissue were significantly increased(P＜0.01).The 1-year,3-year,and 5-year overall survival of the EAC patients in SPP1 low expression group was higher than those in SPP1 high expression group(HR=10.1,P＜0.05;HR=3.09,P＜0.05;HR=2.32,P＜0.05),and the 5-year overall survival of the EAC patients in TGFB1 low expression group was higher than that in TGFB1 high expression group(HR=2.36,P＜0.05).The UALCAN database analysis results showed that compared with esophageal normal tissue,the expression levels of SPP1 and TGFB1 mRNA in cancer tissue of the EAC patients with stage Ⅱ-Ⅲ and N1-N2 lymph node metastasis were significantly increased(P＜0.01).The TIMER analysis results showed that the expression levels of SPP1 and TGFB1 mRNA in cancer tissue of the EAC patients were positively correlated with the infiltration of macrophages(r=0.353,P＜0.01;r=0.187,P＜0.05)and dendritic cells(r=0.236,P＜0.01;r=0.221,P＜0.01).The GO and KEGG pathway enrichment analysis results showed that SPP1,TGFB1,and their top 50 positively correlated genes mainly participated in the biological processes such as cell migration,cell activity,and angiogenesis,and signaling pathways such as tumor proteoglycans,extracellular matrix(ECM)-receptor interaction,and phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT).Conclusion:SPP1 and TGFB1 are closely associated with clinical staging,lymph node metastasis,and overall survival of the EAC patients.High expressions of SPP1 and TGFB1 may lead to the infiltration of the macrophages and dendritic cells,and change the tumor microenvironment.SPP1 and TGFB1 may become new targets for the diagnosis and treatment of EAC.

Objective To explore the role of ferroptosis-related genes in regulating ferroptosis of esophageal squamous cell carcinoma(ESCC).Methods ESCC datasets GSE161533 and GSE20347 were downloaded from the Gene Expression Omnibus(GEO)to identify the differentially expressed genes(DEGs)using R software.ESCC ferroptosis-related genes obtained by intersecting the DEGs with ferroptosis-related genes from FerrDb were analyzed using GO and KEGG analyses,protein-protein interaction(PPI)network analysis,and core gene identification through Cytoscape.The identified ferroptosis suppressor genes were validated using TCGA database,and their expression levels were detected using RT-qPCR in cultured normal esophageal cells and ESCC cells.Six ferroptosis suppressor genes(RRM2,GCLC,TFRC,TXN,SLC7A11,and EZH2)were downregulated with siRNA in ESCC cells,and the changes in cell proliferation and apoptosis were assessed with CCK8 assay and flow cytometry;Western blotting was performed to examine the changes in ferroptosis progression of the cells.Results We identified a total of 58 ESCC ferroptosis-related genes,which involved such biological processes as glutathione transmembrane transport,iron ion transport,and apoptosis and the ferroptosis,glutathione metabolism,and antifolate resistance pathways.The PPI network included 54 nodes and 74 edges with a clustering coefficient of 0.522 and PPI enrichment P<0.001.Cytoscape identified 6 core ferroptosis suppressor genes(RRM2,TFRC,TXN,EZH2,SLC7A11,and GCLC),which were highly expressed in ESCC tissues in the TCGA dataset and in ESCC cell lines.Downregulating these genes in ESCC TE1 cells significantly inhibited cell proliferation,promoted cell apoptosis,reduced the expression levels of ferroptosis markers GPX4 and FIH1,and increased the expression of ACSL4.Conclusion High expression of ferroptosis suppressor genes in ESCC may cause arrest of ferroptosis progression to facilitate tumor development,and inhibiting these genes can restore ferroptosis and promote cell apoptosis,suggesting their value as potential therapeutic targets for ESCC.

Objective To explore the role of ferroptosis-related genes in regulating ferroptosis of esophageal squamous cell carcinoma(ESCC).Methods ESCC datasets GSE161533 and GSE20347 were downloaded from the Gene Expression Omnibus(GEO)to identify the differentially expressed genes(DEGs)using R software.ESCC ferroptosis-related genes obtained by intersecting the DEGs with ferroptosis-related genes from FerrDb were analyzed using GO and KEGG analyses,protein-protein interaction(PPI)network analysis,and core gene identification through Cytoscape.The identified ferroptosis suppressor genes were validated using TCGA database,and their expression levels were detected using RT-qPCR in cultured normal esophageal cells and ESCC cells.Six ferroptosis suppressor genes(RRM2,GCLC,TFRC,TXN,SLC7A11,and EZH2)were downregulated with siRNA in ESCC cells,and the changes in cell proliferation and apoptosis were assessed with CCK8 assay and flow cytometry;Western blotting was performed to examine the changes in ferroptosis progression of the cells.Results We identified a total of 58 ESCC ferroptosis-related genes,which involved such biological processes as glutathione transmembrane transport,iron ion transport,and apoptosis and the ferroptosis,glutathione metabolism,and antifolate resistance pathways.The PPI network included 54 nodes and 74 edges with a clustering coefficient of 0.522 and PPI enrichment P<0.001.Cytoscape identified 6 core ferroptosis suppressor genes(RRM2,TFRC,TXN,EZH2,SLC7A11,and GCLC),which were highly expressed in ESCC tissues in the TCGA dataset and in ESCC cell lines.Downregulating these genes in ESCC TE1 cells significantly inhibited cell proliferation,promoted cell apoptosis,reduced the expression levels of ferroptosis markers GPX4 and FIH1,and increased the expression of ACSL4.Conclusion High expression of ferroptosis suppressor genes in ESCC may cause arrest of ferroptosis progression to facilitate tumor development,and inhibiting these genes can restore ferroptosis and promote cell apoptosis,suggesting their value as potential therapeutic targets for ESCC.

Objective:To investigate the effects of Anhydroicaritin (AHI) , an isopentenylated flavo-noid compound, on proliferation, migration and apoptosis of human hepatocarcinoma cell line MHCC-97H.Methods:Human hepatocarcinoma cell line MHCC-97H and human normal liver cell line L02 were cultured in vitro. MHCC-97H cells were treated with 0, 20, 40, 80, 120, 160, 200 μg/ml of AHI respectively and L02 cells were treated with 0, 25, 50, 100, 150, 200, 400, 500 μg/ml of AHI respectively. CCK-8 and clone formation assay were used to detect cell proliferation. Scratch test was used to explore cell migration ability. Hoechst33342 assay and flow cytometer were used to detect cell apoptosis. The expressions of apoptosis-related proteins were detected by Western blotting. Results:The cell viabilities of MHCC-97H cells treated with 0, 20, 40, 80, 120, 160, 200 μg/ml of AHI for 24 h were (100.00±0.00) %, (97.41±2.10) %, (96.58±3.23) %, (87.72±4.85) %, (78.33±3.76) %, (56.97±2.61) % and (15.25±2.51) % respectively, and there was a statistically significant difference ( F=429.20, P<0.001) . There were statistically significant differences between 0 μg/ml and 80, 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . The cell viabilities of L02 cells treated with 0, 25, 50, 100, 150, 200, 400, 500 μg/ml of AHI for 24 h were (100.00±0.00) %, (96.82±3.79) %, (95.36±3.43) %, (90.79±5.75) %, (77.67±5.66) %, (63.98±5.22) %, (34.22±4.01) % and (33.84±4.41) % respectively, and there was a statistically significant difference ( F=233.20, P<0.001) . There were statistically significant differences between 0 μg/ml and 100, 150, 200, 400, 500 μg/ml of AHI treatment (all P<0.05) . The 24 h half maximal inhibitory concentration (IC 50) value of AHI treated L02 cells was (300.20±17.10) μg/ml, which was significantly higher than that of MHCC-97H cells [ (158.60±5.50) μg/ml], and there was a statistically significant difference ( t=13.65, P<0.001) . The cell clone numbers of MHCC-97H cells treated with 0, 120, 160 and 200 μg/ml of AHI for 24 h were 1 993.00±46.29, 1 355.00±54.84, 998.33±21.03 and 218.33±35.95 respectively, and there was a statistically significant difference ( F=954.80, P<0.001) . There were statistically significant differences between 0 μg/ml and 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . The healing rates of MHCC-97H cells treated with 0, 120, 160 and 200 μg/ml of AHI for 24 h were (51.68±1.93) %, (16.04±0.73) %, (8.88±0.31) % and (-6.94±0.46) % respectively, and there was a statistically significant difference ( F=1 616.00, P<0.001) . There were statistically significant differences between 0 μg/ml and 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . Hoechst33342 experiment showed that MHCC-97H cells treated with 0 μg/ml AHI showed uniform dark blue with a complete nuclear state under inverted microscope. Compared with 0 μg/ml AHI treated cells, cells in the 120, 160, 200 μg/ml AHI treatment groups wrinkled and broken, and nuclei were also morphologically abnormal, with some nuclei stained bright blue, and the situation became more obvious with increasing dose. The apoptosis rates of MHCC-97H cells treated with 0, 120, 160 and 200 μg/ml AHI for 24 h were (10.51±0.56) %, (42.23±0.87) %, (61.92±0.52) % and (72.05±0.74) % respectively, and there was a statistically significant difference ( F=4 677.00, P<0.001) . There were statistically significant differences between 0 μg/ml and 120, 160, 200 μg/ml of AHI treatment (all P<0.001) . There were statistically significant differences among the different expression levels of Bax, Cleaved Caspase-3/Caspase-3, Cleaved Caspase-9/Caspase-9, and Bcl-2 proteins in MHCC-97H cells of 0, 120, 160, and 200 μg/ml of AHI treatment ( F=30.43, P<0.001; F=212.80, P<0.001; F=475.30, P<0.001; F=10.75, P=0.004) . The Bax protein expression of 160 and 200 μg/ml was significantly increased than that of 0 μg/ml AHI (both P<0.001) . The Cleaved Caspase-3/Caspase-3, Cleaved Caspase-9/Caspase-9 protein expressions of 120, 160 and 200 μg/ml were significantly increased than those of 0 μg/ml AHI (all P<0.001) . The Bcl-2 protein expression of 120, 160, 200 μg/ml was significantly decreased compared with that of 0 μg/ml AHI (all P<0.05) . Conclusion:AHI can inhibit the proliferation and migration of hepatocellular carcinoma cell line MHCC-97H, and promote its apoptosis.

The brain pericyte is a unique and indispensable part of the blood-brain barrier (BBB), and contributes to several pathological processes in traumatic brain injury (TBI). However, the cellular and molecular mechanisms by which pericytes are regulated in the damaged brain are largely unknown. Here, we show that the formation of neutrophil extracellular traps (NETs) induces the appearance of CD11b⁺ pericytes after TBI. These CD11b⁺ pericyte subsets are characterized by increased permeability and pro-inflammatory profiles compared to CD11b^- pericytes. Moreover, histones from NETs by Dectin-1 facilitate CD11b induction in brain pericytes in PKC-c-Jun dependent manner, resulting in neuroinflammation and BBB dysfunction after TBI. These data indicate that neutrophil-NET-pericyte and histone-Dectin-1-CD11b are possible mechanisms for the activation and dysfunction of pericytes. Targeting NETs formation and Dectin-1 are promising means of treating TBI.
Blood-Brain Barrier/metabolism* ; Brain/pathology* ; Brain Injuries, Traumatic/metabolism* ; Extracellular Traps/metabolism* ; Histones ; Humans ; Lectins, C-Type ; Pericytes/pathology*

Objective To investigate the expression of Aire,Foxp3,AchR and other immune factors in human thymoma tissue and plasma and explore their role in myasthenia gravis with thymoma.Methods T lymphocyte subsets,immunoglobulin and other immune factors in plasma were compared,and the Expression of Aire,Foxp3 and AchR were examined in thymoma by reverse transcriptional polymerase chain reaction and immunohistochemical staining,and the results were analyzed by SPSS statistics software.Results The ratio of CD4 + to CD8 + T lymphocyte was much higher in plasma,while the expressions of Aire,Foxp3 and AchR at mRNA and protein level were much lower in thymoma patients with myasthenia gravis,and related to Ossermann subtype,WHO subgroup and Masaoka stage.The differences were statistically significant (P ＜ 0.05).Conclusion The ratio of CD4 + to CD8 + T lymphocyte and the abnormal expressions of Aire and Foxp3could used as an indicator of immune state in thymoma patient with myasthenia gravis and play an important role in the development of thymoma with myasthenia gravis,but the mechanism is indefinite.