Objective To investigate the relationship between plasma metalloproteinase-tissue inhibitor 1 (TIMP-1),vascular endothelial growth factor (VEGF) and potentially transforming growth factor binding protein 2 (LTBP-2) levels and risk stratification and death in patients with acute pulmonary embolism (APE). Methods A toral of 110 APE patients admitted to the Third People's Hospital of Haikou from January 2022 to January 2024 were selected for risk stratification,and they were divided into low-risk group(n=28),medium-risk group(n=43) and high-risk groups(n=39). According to the occurrence of death in APE patients,they were divided into a survival group (n=79)and a death group(n=31). Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of plasma TIMP-1,VEGF and LTBP-2. Applying multiple Logistic regression analysis to identify risk factors affecting the mortality of APE patients and plotting ROC curve to analyze the predictive value of plasma TIMP-1,VEGF and LTBP-2 levels for APE patients mortality and. Pearson correlation analysis was used to analyze the correlation between plasma levels of TIMP-1,VEGF and LTBP-2 and clinical indexes in APE patients. Results The heart rate,B-type brain natriuretic peptide (BNP),D-dimer,TIMP-1(207.15±62.84pg/ml vs 152.48±41.62pg/ml),VEGF (726.35±190.46pg/ml vs 419.27±115.28pg/ml) and LTBP-2(29.17±6.38ng/ml vs 13.26±3.70ng/ml) levels in the death group were significantly higher than those in the survival group,the difference were statistically significant (t=5.386～21.194,all P<0.05). The levels of plasma TIMP-1(204.15±60.17 pg/ml,178.18±51.30pg/ml vs 148.20±34.80pg/ml),VEGF(720.83±204.18pg/ml,580.16±158.37pg/ml vs 412.15±109.26pg/ml) and LTBP-2(28.40±6.41ng/ml,21.37±5.26ng/ml vs 12.84±3.12ng/ml) in high-risk and medium-risk groups were significantly higher than those in the low-risk group(t=8.417～19.850),and those in the high-risk group were higher than those in the medium-risk group(t=7.964,9.381,11.470),the differences were statistically significant (all P<0.05),respectively. Multiple Logistic regression analysis showed that increased plasma BNP,D-dimer,TIMP-1,VEGF and LTBP-2 levels were risk factors for death in APE patients (all P<0.05). ROC curve analysis showed that TIMP-1,VEGF and LTBP-2 combined predicted the highest AUC(95％CI) of death in APE patients[0.938(0.881～0.997)],with an accuracy of 88.2％. Pearson correlation analysis showed that the levels of plasma TIMP-1,VEGF and LTBP-2 in APE patients were positively correlated with BNP and D-dimer (r=0.416～0.753,all P<0.05). Conclusion Elevated levels of plasma TIMP-1,VEGF and LTBP-2 are associated with high risk and mortality in APE,and the combination of these three factors has good predictive value for mortality in APE patients.

Objective To identify genetic regulators of ionizing radiation(IR)sensitivity through clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated nuclease 9(Cas9)genome-wide screening.Methods A specialized single guide RNA(sgRNA)library was developed targeting 987 stress-response regulatory genes identified through Kyoto Encyclopedia of Genes and Genomes(KEGG),Reactome and gene ontology(GO)databases,followed by construction of sgRNA plasmids and packaging into a lentiviral library.Using colon adenocarcinoma Caco-2 cells as a model system,the Cas9-stable transgenic line was established via lentiviral transduction and antibiotic selection.Library-transduced cells underwent puromycin selection,followed by γ-irradiation(dose optimized via preliminary experiments).Post-irradiation phenotypic selection was conducted after 14 days,with subsequent next-generation sequencing of surviving cell populations to identify enriched/depleted sgRNAs.Candidate genes were functionally validated through orthogonal assays:cell counting kit-8(CCK-8)proliferation analysis,clonogenic survival assays and flow cytometric quantification of reactive oxygen species(ROS)and apoptotic markers.Results The optimized screening platform identified 281 radiation response genes(165 radiosensitive and 116 radioprotective candidates).Functional validation revealed Bcl2-associated athanogene 3(BAG3)knockdown significantly enhanced radioresistance.Proliferation was increased 1.2-1.5 fold,clonogenic survival improved 1.5-2.0 fold,and ROS was reduced by 13％-25％coupled with 32％-73％apoptosis attenuation compared to control groups.Conclusion The integrated CRISPR/Cas9 screening platform effectively identifies novel radiation response regulators,as evidenced by the discovery of BAG3 as a critical radiosensitivity determinant.This high-throughput functional genomics approach provides a robust methodology for systematically mapping molecular determinants of cellular radiation response,with potential applications in both mechanistic studies and therapeutic target discovery.

Objective To investigate the relationship between plasma metalloproteinase-tissue inhibitor 1 (TIMP-1),vascular endothelial growth factor (VEGF) and potentially transforming growth factor binding protein 2 (LTBP-2) levels and risk stratification and death in patients with acute pulmonary embolism (APE). Methods A toral of 110 APE patients admitted to the Third People's Hospital of Haikou from January 2022 to January 2024 were selected for risk stratification,and they were divided into low-risk group(n=28),medium-risk group(n=43) and high-risk groups(n=39). According to the occurrence of death in APE patients,they were divided into a survival group (n=79)and a death group(n=31). Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of plasma TIMP-1,VEGF and LTBP-2. Applying multiple Logistic regression analysis to identify risk factors affecting the mortality of APE patients and plotting ROC curve to analyze the predictive value of plasma TIMP-1,VEGF and LTBP-2 levels for APE patients mortality and. Pearson correlation analysis was used to analyze the correlation between plasma levels of TIMP-1,VEGF and LTBP-2 and clinical indexes in APE patients. Results The heart rate,B-type brain natriuretic peptide (BNP),D-dimer,TIMP-1(207.15±62.84pg/ml vs 152.48±41.62pg/ml),VEGF (726.35±190.46pg/ml vs 419.27±115.28pg/ml) and LTBP-2(29.17±6.38ng/ml vs 13.26±3.70ng/ml) levels in the death group were significantly higher than those in the survival group,the difference were statistically significant (t=5.386～21.194,all P<0.05). The levels of plasma TIMP-1(204.15±60.17 pg/ml,178.18±51.30pg/ml vs 148.20±34.80pg/ml),VEGF(720.83±204.18pg/ml,580.16±158.37pg/ml vs 412.15±109.26pg/ml) and LTBP-2(28.40±6.41ng/ml,21.37±5.26ng/ml vs 12.84±3.12ng/ml) in high-risk and medium-risk groups were significantly higher than those in the low-risk group(t=8.417～19.850),and those in the high-risk group were higher than those in the medium-risk group(t=7.964,9.381,11.470),the differences were statistically significant (all P<0.05),respectively. Multiple Logistic regression analysis showed that increased plasma BNP,D-dimer,TIMP-1,VEGF and LTBP-2 levels were risk factors for death in APE patients (all P<0.05). ROC curve analysis showed that TIMP-1,VEGF and LTBP-2 combined predicted the highest AUC(95％CI) of death in APE patients[0.938(0.881～0.997)],with an accuracy of 88.2％. Pearson correlation analysis showed that the levels of plasma TIMP-1,VEGF and LTBP-2 in APE patients were positively correlated with BNP and D-dimer (r=0.416～0.753,all P<0.05). Conclusion Elevated levels of plasma TIMP-1,VEGF and LTBP-2 are associated with high risk and mortality in APE,and the combination of these three factors has good predictive value for mortality in APE patients.