Objective To observe the value of three-dimensional speckle tracking imaging(3D-STI)in evaluation on right ventricle(RV)function for predicting adverse events in patients with unstable angina(UA)after percutaneous coronary intervention(PCI).Methods A total of 155 UA patients who underwent both 3D-STI and conventional echocardiography and planned to accept PCI were prospectively enrolled and divided into adverse event group(n=37)and control group(n=118)based on the occurrence of adverse events after PCI or not.Clinical data and echocardiographic parameters were compared between groups,and new binary variables were generated according to the optimal cut-off values showed by the maximum Youden index of receiver operating characteristic curves of 3D-STI parameters being statistically different between groups.Then the independent predictors of adverse events among UA patients after PCI were analyzed,a combined model was constructed,and its predictive value was assessed.Results In adverse event group,the left ventricular end diastolic volume(LVEDV)was higher,while the left ventricular ejection fraction(LVEF),right ventricular free wall longitudinal strain(RVFWLS)and right ventricular global longitudinal strain(RVGLS)were all lower than those in control group(all P＜0.05).The optimal cut-off values for distinguishing adverse event or not was 110 ml for LVEDV,19.05%for RVFWLS and 19.75%for RVGLS,respectively,and all the 3 generated binary variables were independent predictors for adverse events of UA patients after PCI(all P＜0.05).The area under the curve of combined model for predicting adverse events in UA patients after PCI was 0.864.Conclusion RVFWLS＜19.05%and RVGLS＜19.75%obtained with 3D-STI,as well as LVEDV＞110 ml with conventional echocardiography were all independent predictors of adverse events in UA patients after PCI.

Although no cross protection was observed between different serotypes of foot-and-mouth disease virus(FMDV),there were cross-reactivity between different serotypes of antibodies produced after vaccination,the aim of this paper was to prepare the neutralizing monoclonal anti-body against Foot-and-mouth disease virus(FMDV)serotype O,and to develop the method to dis-tinguish antibody against FMDV serotype O and A based on mAb.The inactivated FMDV serotype O was used as antigen in mAb production,a series of GST fusion overlapping peptides and trun-cated peptides expressed in Escherichia coli were used to identify antigenic epitope recognized by monoclonal antibodies.In order to verify feasibility of the screened monoclonal antibodies in diag-nosis,20 positive serum of FMDV serotype O and A,20 negative serum with known background were detected by blocking ELISA.Results were as follows:five monoclonal antibodies were suc-cessfully screened.The five monoclonal antibodies showed good reactivity with FMDV serotype O,but did not react with FMDV serotype A by Western blot and IFA,these mAbs showed neutrali-zing ability to FMDV/O/MY98/GZBY/2013 by VNT.The same epitope was identified by five monoclonal antibodies,the minimum epitope was145 RGDLQVLA152,Arg145 and Gln149 were key a-mino acids of the epitope.Sequence alignment analysis revealed that the identified epitopes were conserved among most of O type FMDV strains,but Gln149 was mutated among all A,Asia 1 and SAT1-3 type FMDV strains.The mAb-8C5D3 distinguished between antibody of FMDV serotype O and FMDV serotype A by blocking ELISA.The results provided materials for development of O type FMDV antibody detection kit and evaluation of vaccine immune effect.

Objective:To investigate the role of aging-related genes(ARGs)in the prognosis of lung squamous cell carcinoma(LUSC)and establish a novel prognostic prediction model.Methods:Transcriptomic data and clinical information of LUSC patients were obtained from TCGA,combined with ARGs from Aging Atlas.Key genes were screened through differential expression analysis,survival analysis,and Cox regression to construct a prognostic model.Model performance was validated in clinical subgroups,and biological pathway enrichment(GSEA)and immune microenvironment analyses were performed.Results:Five ARGs(ERFFI1,MDH1,SENP2,SNAI1,TP63)were identified to build the model.Significant survival differences were observed between high-and low-risk groups(P＜0.001),with 1-,3-,and 5-year AUC values of 0.610,0.668,and 0.665,respectively.The risk score was an independent prognostic factor(HR=11.261,95％CI:3.654-34.701,P＜0.001)and showed predictive efficacy in both early-stage(Ⅰ-Ⅱ,P=0.022)and advanced-stage(Ⅲ-Ⅳ,P=0.004)patients.GSEA revealed significant enrichment of Alzheimer's disease(P=0.003)and cell adhesion pathways(P=0.008)in high-risk groups.SNAI1 correlated positively with M1/M2 macrophage infiltration(r=0.45,P＜0.001),MDH1 associated with 12 immune cell types(|r|＞0.3,P＜0.05),and the risk score linked to CD8+T cells(r=0.38)and M2 macrophages(r=0.32)(both P＜0.001).Twenty-three immune checkpoints(e.g.,TNFRSF14,CD200R1)were differentially expressed between groups and survival-related(P＜0.05).High-risk patients exhibited elevated TIDE scores(P＜0.001),indicating enhanced immune suppression.Conclusion:This model provides a novel tool for LUSC prognosis assessment,but further clinical validation is required.

Although no cross protection was observed between different serotypes of foot-and-mouth disease virus(FMDV),there were cross-reactivity between different serotypes of antibodies produced after vaccination,the aim of this paper was to prepare the neutralizing monoclonal anti-body against Foot-and-mouth disease virus(FMDV)serotype O,and to develop the method to dis-tinguish antibody against FMDV serotype O and A based on mAb.The inactivated FMDV serotype O was used as antigen in mAb production,a series of GST fusion overlapping peptides and trun-cated peptides expressed in Escherichia coli were used to identify antigenic epitope recognized by monoclonal antibodies.In order to verify feasibility of the screened monoclonal antibodies in diag-nosis,20 positive serum of FMDV serotype O and A,20 negative serum with known background were detected by blocking ELISA.Results were as follows:five monoclonal antibodies were suc-cessfully screened.The five monoclonal antibodies showed good reactivity with FMDV serotype O,but did not react with FMDV serotype A by Western blot and IFA,these mAbs showed neutrali-zing ability to FMDV/O/MY98/GZBY/2013 by VNT.The same epitope was identified by five monoclonal antibodies,the minimum epitope was145 RGDLQVLA152,Arg145 and Gln149 were key a-mino acids of the epitope.Sequence alignment analysis revealed that the identified epitopes were conserved among most of O type FMDV strains,but Gln149 was mutated among all A,Asia 1 and SAT1-3 type FMDV strains.The mAb-8C5D3 distinguished between antibody of FMDV serotype O and FMDV serotype A by blocking ELISA.The results provided materials for development of O type FMDV antibody detection kit and evaluation of vaccine immune effect.

Objective:To investigate the role of aging-related genes(ARGs)in the prognosis of lung squamous cell carcinoma(LUSC)and establish a novel prognostic prediction model.Methods:Transcriptomic data and clinical information of LUSC patients were obtained from TCGA,combined with ARGs from Aging Atlas.Key genes were screened through differential expression analysis,survival analysis,and Cox regression to construct a prognostic model.Model performance was validated in clinical subgroups,and biological pathway enrichment(GSEA)and immune microenvironment analyses were performed.Results:Five ARGs(ERFFI1,MDH1,SENP2,SNAI1,TP63)were identified to build the model.Significant survival differences were observed between high-and low-risk groups(P＜0.001),with 1-,3-,and 5-year AUC values of 0.610,0.668,and 0.665,respectively.The risk score was an independent prognostic factor(HR=11.261,95％CI:3.654-34.701,P＜0.001)and showed predictive efficacy in both early-stage(Ⅰ-Ⅱ,P=0.022)and advanced-stage(Ⅲ-Ⅳ,P=0.004)patients.GSEA revealed significant enrichment of Alzheimer's disease(P=0.003)and cell adhesion pathways(P=0.008)in high-risk groups.SNAI1 correlated positively with M1/M2 macrophage infiltration(r=0.45,P＜0.001),MDH1 associated with 12 immune cell types(|r|＞0.3,P＜0.05),and the risk score linked to CD8+T cells(r=0.38)and M2 macrophages(r=0.32)(both P＜0.001).Twenty-three immune checkpoints(e.g.,TNFRSF14,CD200R1)were differentially expressed between groups and survival-related(P＜0.05).High-risk patients exhibited elevated TIDE scores(P＜0.001),indicating enhanced immune suppression.Conclusion:This model provides a novel tool for LUSC prognosis assessment,but further clinical validation is required.

Objective To observe the value of three-dimensional speckle tracking imaging(3D-STI)in evaluation on right ventricle(RV)function for predicting adverse events in patients with unstable angina(UA)after percutaneous coronary intervention(PCI).Methods A total of 155 UA patients who underwent both 3D-STI and conventional echocardiography and planned to accept PCI were prospectively enrolled and divided into adverse event group(n=37)and control group(n=118)based on the occurrence of adverse events after PCI or not.Clinical data and echocardiographic parameters were compared between groups,and new binary variables were generated according to the optimal cut-off values showed by the maximum Youden index of receiver operating characteristic curves of 3D-STI parameters being statistically different between groups.Then the independent predictors of adverse events among UA patients after PCI were analyzed,a combined model was constructed,and its predictive value was assessed.Results In adverse event group,the left ventricular end diastolic volume(LVEDV)was higher,while the left ventricular ejection fraction(LVEF),right ventricular free wall longitudinal strain(RVFWLS)and right ventricular global longitudinal strain(RVGLS)were all lower than those in control group(all P＜0.05).The optimal cut-off values for distinguishing adverse event or not was 110 ml for LVEDV,19.05%for RVFWLS and 19.75%for RVGLS,respectively,and all the 3 generated binary variables were independent predictors for adverse events of UA patients after PCI(all P＜0.05).The area under the curve of combined model for predicting adverse events in UA patients after PCI was 0.864.Conclusion RVFWLS＜19.05%and RVGLS＜19.75%obtained with 3D-STI,as well as LVEDV＞110 ml with conventional echocardiography were all independent predictors of adverse events in UA patients after PCI.

Objective To observe the risk factors of ischemic mitral regurgitation(IMR)in ischemic cardiomyopathy.Methods Totally 143 patients with ischemic cardiomyopathy were retrospectively enrolled and divided into IMR+group(n=68)or IMR-group(n=75)based on IMR,while 50 healthy volunteers were taken as controls(control group).The general information,conventional ultrasonic parameters of left ventricle,three-dimensional speckle tracking imaging(3D-STI)parameters as well as mitral valve structural and functional parameters were compared among groups,and the risk factors of IMR were screened with logistic regression analysis.Results Significant differences of left ventricular end-diastolic volume(LVEDV),left ventricular end-systolic volume(LVESV),left ventricular global longitudinal strain(LVGLS),the peak systolic twist(Twist),tenting volume(VTent)and total leaflet area(TLA)/annulus area(AA)were found between IMR+group and IMR-group(all P＜0.05).Decreased LVGLS,decreased Twist and increased VTent were all independent risk factors of IMR in ischemic cardiomyopathy(all P＜0.05).Conclusion Decreased LVGLS,decreased Twist and increased VTent were independent risk factors of IMR in ischemic cardiomyopathy.

Coronary perforation is when a contrast agent or blood flows outside a blood vessel through a tear in a coronary artery.In this case,we reported a case of percutaneous coronary intervention for coronary calcified lesions,which led to iatrogenic coronary perforation and cardiac tamponade after the use of Shockwave balloon to treat intracoronary calcified nodules,and the management of PCI-related CAP was systematically reviewed through the literature.

A prokaryotic expression vector for the mutant diphtheria toxin CRM197 was constructed and expressed in Esch-erichia coli cells.Anti-CRM197 antibody concentrations were detected in serum samples of healthy volunteers.The crm 197 gene was codon-optimized in E.coli and cloned into the plasmid pET28a(+)under optimized expression conditions.CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography,and confirmed by western blot analysis.The puri-fied CRM197 was used to detect specific anti-CRM197 antibody levels in serum samples of different age groups.The results showed that soluble codon-optimized CRM197 was successfully expressed under optimized expression conditions.The purity of CRM197 was more than 95％,as determined with Ni-NTA spin columns and ion exchange chromatography,consistent with the single specific bands obtained by western blot analysis and detection of serum levels of the anti-CRM197 antibody.Collec-tively,these results confirmed that the proposed expression strategy achieved high-yield production of soluble CRM197,al-though high levels in human serum may affect evaluation of immune interactions with glycan-CRM197 conjugates for applica-tion as a diagnostic antigen.The diphtheria mutant toxin CRM197 is used in many conjugate vaccines.The synthetic crm 197 gene with codon optimization in pET28a was transformed into E.coli Origami B(DE3)cells.CRM197 was induced by isopro-pyl β-d-1-thiogalactopyranoside and high level accumulation of soluble CRM197 was purified using Ni-NTA spin columns and ion exchange chromatography.The purity of the final prepara-tion reached 95％.CRM197 was used to detect the concentra-tions of the anti-CRM197 antibody in serum samples of healthy volunteers of different ages.The proposed expression strategy yielded high production of CRM197,which could interfere with evaluations of induced immune interactions by glycan-CRM197 conjugates and prohibit application as a diagnostic antigen.

Objective To propose a classification method for ultrasound images of pedicle screw canals based on radiomics analysis,and to evaluate the integrity of the screw canal.Methods With thoracolumbar spine specimens from 4 fresh cadavers,50 pedicle screw canals were pre-established and ultrasound images of the canals were acquired.A total of 2 000 images(1 000 intact and 1 000 damaged canal samples)were selected.The dataset was randomly divided in a 4∶1 ratio using 5-fold cross-validation to form training and testing sets(consisting of 1 600 and 400 samples,respectively).Firstly,the optimal radius of the region of interest was identified using the Otsu's thresholding method,followed by feature extraction using pyradiomics.Principal component analysis and the least absolute shrinkage and selection operator algorithm were employed for dimensionality reduction and feature selection,respectively.Subsequently,3 machine learning models(support vector machine[SVM],logistic regression,and random forest)and 3 deep learning models(visual geometry group[VGG],ResNet,and Transformer)were used to classify the ultrasound images.The performance of each model was evaluated using accuracy.Results With a region of interest radius of 230 pixels,the SVM model achieved the highest classification accuracy of 96.25%.The accuracy of the VGG model was only 51.29%,while the accuracies of the logistic regression,random forest,ResNet,and Transformer models were 85.50%,80.75%,80.17%,and 75.18%,respectively.Conclusion For ultrasound images of pedicle screw canals,the machine learning model performs better than the deep learning model as a whole,and the SVM model has the best classification performance,which can be used to assist physicians in diagnosis.