Traditional Chinese medicine contains a variety of volatile components （VOCs） such as terpenes and aliphatics, which have significant pharmacological activities, and are commonly analyzed by gas chromatography （GC） and gas chromatography-mass spectrometry （GC-MS）. Although the separation ability is strong, the samples need to be pre-treated and the analysis time is long. In recent years, the combination of GC and ion migration spectrometry （IMS） has been widely used in the academic community. The combination of high separation ability of GC and rapid reaction ability of IMS has the advantages of no sample pretreatment, fast analysis time, low detection limit and simple operation. It is increasingly used in traditional Chinese medicine research. The working principle of GC-IMS was introduced, the application in VOCs research of traditional Chinese medicine was summarized, and the future development direction was prospected in this paper.

Objective To investigate the expression of Delta(24)-cholesteryl reductase(DHCR24)proteins in cervical squamous intraepithelial lesions(SILs)tissues and its value in different cervical lesion patho-logical diagnosis.Methods The expression of DHCR24,p16,and Ki-67 was quantitatively detected by immuno-histochemistry in 51 normal cervical tissues,44 LSILs,and 57 HSILs.The receiver operating characteristic(ROC)curve was drawn to analyze the diagnostic efficacy of DHCR24,p16,and Ki-67 proteins in evaluating the degree of SILs.Results The expression levels of DHCR24,p16 and Ki-67 protein were positively correlated with the progression of SILs(P<0.05).ROC analysis showed that the immunohistochemistry score cutoff value for DHCR24 between normal cervical tissue and LSIL was 0.1145,and between LSIL and HSIL was 0.1969.The sensi-tivity of DHCR24 in diagnosing LSIL was 79.55%,higher than that for p16 and Ki-67,which was 15.91%and 18.08%(P<0.05).The area under the ROC curve(AUC)for distinguishing normal cervical tissue from LSIL using a combination of DHCR24 and p16 was 0.932(95%CI:0.878～0.986),higher than that for p16 and Ki-67 combined,which was 0.861(95%CI:0.785～0.936).The AUC for distinguishing LSIL from HSIL using a combi-nation of DHCR24 and p16 was0.971(95%CI:0.946～0.997),higher than that for p16 and Ki-67 combined,which was 0.870(95%CI:0.790～0.949).Conclusions Both DHCR24 and p16 protein expression levels can pro-vide reference for the grading of SILs,and their combination can improve the diagnostic efficiency.The cutoff value derived from the ROC curve plotted by DHCR24 immunohistochemical staining intensity can improve the sensitivity of LSIL diagnosis.

Objective To explore the effects of different modes of cerebellar transcranial magnetic stimulation(TMS)on response inhibition function in healthy adults and the underlying mechanisms.Methods Eighty-one healthy adult volunteers were randomly divided into 5 Hz repetitive TMS(rTMS)group(n=28),intermittent theta burst stimulation(iTBS)group(n=26)and sham stimulation group(n=27).The TMS stimulation target was the cerebellar Crus Ⅱ region,and the stimulation site was accurately located by the neuronavigation system.The stimulation intensity was 80％of the individual resting motor threshold(RMT).The Go/No-Go task response time(RT)and response inhibition ability and synchronous EEG were measured before and after intervention in each group.The changes of N2 wave and time-frequency in the time window related to response inhibition function in the event-related potential(ERP)were analyzed.Results Compared with that pre-stimulation,RT after stimulation in rTMS group and iTBS group was significantly shortened(all P＜0.05).There was a significant difference in the difference of RT before and after stimulation among the three groups(P＜0.001).Compared with that in the sham stimulation group,the RT after stimulation in the rTMS group and the iTBS group was significantly shortened(P＜0.05-0.01).There was no significant difference in RT changes before and after stimulation between rTMS group and iTBS group(P＞0.05).There was no significant difference in the correct rate of Go task and No-Go task before and after stimulation in the rTMS group,iTBS group and sham stimulation group(all P＞0.05).There was no significant difference in the changes of correct rate in Go task and No-Go task among the three groups(P＞0.05).In the No-Go condition,compared with pre-TMS intervention,the N2 amplitude at Fz and FCz electrodes in rTMS group and iTBS group significantly decreased after intervention(all P＜0.05).Compared with that in the sham stimulation group,the changes of N2 amplitude in the frontal electrodes of the rTMS group and the iTBS group were statistically significant(all P＜0.01),but there was no significant difference in the N2 amplitude changes before and after intervention between the rTMS and iTBS groups(all P＞0.05).In the Go condition,there was no significant difference in N2 amplitude before and after intervention in each group(all P＞0.05).There was no significant difference in the latency changes of Go/No-Go task before and after intervention in each group(all P＞0.05).In the No-Go task,the theta frequency power was increased significantly after rTMS intervention(P＜0.01),and the alpha,beta and gamma frequency powers were decreased significantly(all P＜0.05).Theta frequency power at the Fz channel in the iTBS group was significantly increased after intervention(P＜0.05).No significant time-frequency changes were observed in the sham group before and after the intervention.There were no significant differences in the frequency changes(alpha,theta,beta,gamma)between the rTMS and iTBS groups after intervention(all P＞0.05).In the Go task,there was no significant change in each frequency before and after intervention in each group.Correlation analysis showed that there was no significant correlation between RT and N2 amplitude and theta oscillation activity in each group(all P＞0.05).Conclusions Both 5 Hz rTMS and iTBS interventions significantly improve response inhibition ability in healthy adults.The two types of stimulation may modulate cerebellar executive control functions through similar neuroregulatory mechanisms,providing new evidence for the clinical application of cerebellar TMS in cognitive control-related disorders.

Objective To investigate the expression of Delta(24)-cholesteryl reductase(DHCR24)proteins in cervical squamous intraepithelial lesions(SILs)tissues and its value in different cervical lesion patho-logical diagnosis.Methods The expression of DHCR24,p16,and Ki-67 was quantitatively detected by immuno-histochemistry in 51 normal cervical tissues,44 LSILs,and 57 HSILs.The receiver operating characteristic(ROC)curve was drawn to analyze the diagnostic efficacy of DHCR24,p16,and Ki-67 proteins in evaluating the degree of SILs.Results The expression levels of DHCR24,p16 and Ki-67 protein were positively correlated with the progression of SILs(P<0.05).ROC analysis showed that the immunohistochemistry score cutoff value for DHCR24 between normal cervical tissue and LSIL was 0.1145,and between LSIL and HSIL was 0.1969.The sensi-tivity of DHCR24 in diagnosing LSIL was 79.55%,higher than that for p16 and Ki-67,which was 15.91%and 18.08%(P<0.05).The area under the ROC curve(AUC)for distinguishing normal cervical tissue from LSIL using a combination of DHCR24 and p16 was 0.932(95%CI:0.878～0.986),higher than that for p16 and Ki-67 combined,which was 0.861(95%CI:0.785～0.936).The AUC for distinguishing LSIL from HSIL using a combi-nation of DHCR24 and p16 was0.971(95%CI:0.946～0.997),higher than that for p16 and Ki-67 combined,which was 0.870(95%CI:0.790～0.949).Conclusions Both DHCR24 and p16 protein expression levels can pro-vide reference for the grading of SILs,and their combination can improve the diagnostic efficiency.The cutoff value derived from the ROC curve plotted by DHCR24 immunohistochemical staining intensity can improve the sensitivity of LSIL diagnosis.

Objective Biomarkers associated with liver cancer CD8+T cells identified and prognostic models established by single-cell RNA sequencing and Bulk RNA sequencing.Methods Single-cell datasets of liver cancer were downloaded from the GEO database and differentially expressed genes in CD8+T cells between patients and controls were extracted by single-cell RNA sequencing.Gene expression profiling data and clinical data of liver cancer were downloaded from the TCGA database,and modular genes with relevance to CD8+T cells were screened using the CIBERSORT algorithm and WGCNA technology.Differential genes and modular genes were taken as intersecting genes and subjected to GO and KEGG analyses,and a prognostic model was established by applying univariate COX regression analysis and LASSO algorithm.The predictive effectiveness of the model in both internal and external datasets is validated by K-M and ROC curves.The effectiveness of the model's prediction in both internal and external datasets was validated by K-M and ROC curves.High-and low-risk groups were divided according to the median value of the risk score,and the distribution of infiltrating immune cells and tumor mutations between high-and low-risk groups were analyzed.Results A prognostic model with nine genes was constructed,and the K-M and ROC curves showed that the model had good predictive ability in both the internal and external datasets,and there were significant differences in the distribution of infiltrating immune cells and gene mutations in the high-and low-risk groups.Conclusion In this study,a novel prognostic model based on CD8+T cells was developed using bioinformatic method in combination with single-cell RNA sequencing and Bulk RNA sequencing technologies,which provides a reliable theoretical basis for prognostic improvement and survival prediction of liver cancer patients.

Objective To explore the effects of different modes of cerebellar transcranial magnetic stimulation(TMS)on response inhibition function in healthy adults and the underlying mechanisms.Methods Eighty-one healthy adult volunteers were randomly divided into 5 Hz repetitive TMS(rTMS)group(n=28),intermittent theta burst stimulation(iTBS)group(n=26)and sham stimulation group(n=27).The TMS stimulation target was the cerebellar Crus Ⅱ region,and the stimulation site was accurately located by the neuronavigation system.The stimulation intensity was 80％of the individual resting motor threshold(RMT).The Go/No-Go task response time(RT)and response inhibition ability and synchronous EEG were measured before and after intervention in each group.The changes of N2 wave and time-frequency in the time window related to response inhibition function in the event-related potential(ERP)were analyzed.Results Compared with that pre-stimulation,RT after stimulation in rTMS group and iTBS group was significantly shortened(all P＜0.05).There was a significant difference in the difference of RT before and after stimulation among the three groups(P＜0.001).Compared with that in the sham stimulation group,the RT after stimulation in the rTMS group and the iTBS group was significantly shortened(P＜0.05-0.01).There was no significant difference in RT changes before and after stimulation between rTMS group and iTBS group(P＞0.05).There was no significant difference in the correct rate of Go task and No-Go task before and after stimulation in the rTMS group,iTBS group and sham stimulation group(all P＞0.05).There was no significant difference in the changes of correct rate in Go task and No-Go task among the three groups(P＞0.05).In the No-Go condition,compared with pre-TMS intervention,the N2 amplitude at Fz and FCz electrodes in rTMS group and iTBS group significantly decreased after intervention(all P＜0.05).Compared with that in the sham stimulation group,the changes of N2 amplitude in the frontal electrodes of the rTMS group and the iTBS group were statistically significant(all P＜0.01),but there was no significant difference in the N2 amplitude changes before and after intervention between the rTMS and iTBS groups(all P＞0.05).In the Go condition,there was no significant difference in N2 amplitude before and after intervention in each group(all P＞0.05).There was no significant difference in the latency changes of Go/No-Go task before and after intervention in each group(all P＞0.05).In the No-Go task,the theta frequency power was increased significantly after rTMS intervention(P＜0.01),and the alpha,beta and gamma frequency powers were decreased significantly(all P＜0.05).Theta frequency power at the Fz channel in the iTBS group was significantly increased after intervention(P＜0.05).No significant time-frequency changes were observed in the sham group before and after the intervention.There were no significant differences in the frequency changes(alpha,theta,beta,gamma)between the rTMS and iTBS groups after intervention(all P＞0.05).In the Go task,there was no significant change in each frequency before and after intervention in each group.Correlation analysis showed that there was no significant correlation between RT and N2 amplitude and theta oscillation activity in each group(all P＞0.05).Conclusions Both 5 Hz rTMS and iTBS interventions significantly improve response inhibition ability in healthy adults.The two types of stimulation may modulate cerebellar executive control functions through similar neuroregulatory mechanisms,providing new evidence for the clinical application of cerebellar TMS in cognitive control-related disorders.

Objective To investigate the neurovascular coupling(NVC)status in the acute phase of patients with minor ischemic stroke(MIS)or transient ischemic attack(TIA)due to intracranial large artery moderate-to-severe stenosis or occlusion using multimodal MRI techniques and to explore its correlation with quality of life(QoL).Methods This prospective,consecutive study enrolled patients with MIS or TIA due to intracranial large artery moderate-to-severe stenosis or occlusion form the Department of Neurology,the First Affiliated Hospital of Xi'an Jiaotong University,between June 2022 and October 2023.Recruit healthy subjects with matched age,sex,and handedness form the community during the same period.Patients were divided into left-sided involvement and right-sided involvement groups based on the affected side of the responsible vessel,while the healthy subjects were set as the healthy control group.Post-hoc power analysis was performed using G*Power 3.1 software.General characteristics(age,gender,body mass index,education level)were collected and compared across all three groups.Clinical data and QoL assessment were collected and compared between the two patient groups.Collected clinical data including type of cerebrovascular events(TIA,MIS),the National Institutes of Health stroke scale(NIHSS)score at admission,the responsible vessel(internal carotid artery,middle cerebral artery)and its side location,the degree of responsibility artery stenosis(moderate-severe stenosis[50%-99%stenosis rate],occlusion[100%stenosis rate]),the intracranial collateral circulation status(American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology[ASITN/SIR]collateral circulation grading),cerebrovascular risk factors(hypertension,diabetes,hyperlipidemia,smoking history),and the laboratory test indicators at admission(glycated hemoglobin,triglycerides,total cholesterol,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,blood uric acid,blood homocysteine).QoL was assessed using the stroke impact scale(SIS),covering eight functional domains and a patient-reported overall recovery item.Multimodal MR data were acquired for all subjects.Whole-brain cerebral blood flow(CBF)images were generated using statistics parameter mapping 12(SPM 12)software,while regional homogeneity(ReHo)images were generated using DPABI software.The voxel-wise ratio of CBF to ReHo(CBF/ReHo)was calculated as the regional NVC parameter.Differences in regional NVC characteristics were compared between patient groups and the healthy control group.Correlations between NVC parameters and SIS scores within patient groups were explored.Results(1)A total of 38 patients with MIS or TIA due to intracranial large artery moderate-to-severe stenosis or occlusion were included(26 males,12 females,aged 36-69 years,with mean age of[52±11]years),with 23 in the left-sided involvement group and 15 in the right-sided involvement group.Nineteen healthy subjects were included(10 males,9 females,aged 37-67 years,with mean age of[53±10]years).Post-hoc power analysis showed statistical power of 0.808 for comparing the left-sided involvement group with the healthy control group and 0.762 for comparing the right-sided involvement group with control group.(2)No statistically significant differences were found on gender,age,education level,or body mass index across the three groups(all P＞0.05).No statistically significant differences were observed on the type of cerebrovascular event,cerebrovascular risk factors,distribution of the responsible vessel,degree of stenosis in the responsible vessel,admission NIHSS score,or laboratory test results between the two patient groups(all P＞0.05).There were no statistically significant differences in the total SIS score and the scores of subscales between the two patient groups(all P＞0.05).(3)Compared with the healthy control group,the left-sided involvement group exhibited reduced CBF/ReHo values in the left superior and middle temporal gyri,supramarginal gyrus,middle and inferior frontal gyri,precentral gyrus,angular gyrus,postcentral gyrus,insula,and posterior cerebellar lobe(FDR-corrected,all P＜0.05).In the right-sided involvement group,reduced CBF/ReHo values were observed in the right supramarginal gyrus,right postcentral gyrus,inferior temporal gyrus,and insula(FDR-corrected,all P＜0.05).(4)Correlation analysis revealed that the SIS total score in the left-sided involvement group negatively correlated with CBF/ReHo values in the right inferior frontal gyrus(T=-5.91)and the right middle temporal gyrus(T=-6.65,FDR-corrected,both P＜0.05).The SIS subscale score for activities of daily living in the left-sided involvement group showed negative correlations with CBF/ReHo values in the right angular gyrus(T=-7.36),right medial superior frontal gyrus(T=-6.97),right orbitofrontal cortex(T=-8.99),and left thalamus(T=-7.51,FDR-corrected,all P＜0.05).No significant correlation was observed between the SIS total score and CBF/ReHo values in patients with right-sided involvement group.The SIS subscale for communication score in the right-sided involvement group correlated with CBF/ReHo in the left lingual gyrus(T=-12.15),left olfactory cortex(T=-7.68),and right anterior cingulate and paracingulate cortex(T=-9.46,FDR-corrected,all P＜0.05).Conclusions Patients with MIS or TIA due to intracranial large artery moderate-to-severe stenosis or occlusion show abnormal NVC in the acute phase,especially those with left hemisphere involvement,who exhibit more extensive impairments.QoL in left-sided involvement patients is strongly linked to NVC in the right orbitofrontal cortex and right middle temporal gyrus.These findings require further validation in larger-scale studies.

Objective To investigate the neurovascular coupling(NVC)status in the acute phase of patients with minor ischemic stroke(MIS)or transient ischemic attack(TIA)due to intracranial large artery moderate-to-severe stenosis or occlusion using multimodal MRI techniques and to explore its correlation with quality of life(QoL).Methods This prospective,consecutive study enrolled patients with MIS or TIA due to intracranial large artery moderate-to-severe stenosis or occlusion form the Department of Neurology,the First Affiliated Hospital of Xi'an Jiaotong University,between June 2022 and October 2023.Recruit healthy subjects with matched age,sex,and handedness form the community during the same period.Patients were divided into left-sided involvement and right-sided involvement groups based on the affected side of the responsible vessel,while the healthy subjects were set as the healthy control group.Post-hoc power analysis was performed using G*Power 3.1 software.General characteristics(age,gender,body mass index,education level)were collected and compared across all three groups.Clinical data and QoL assessment were collected and compared between the two patient groups.Collected clinical data including type of cerebrovascular events(TIA,MIS),the National Institutes of Health stroke scale(NIHSS)score at admission,the responsible vessel(internal carotid artery,middle cerebral artery)and its side location,the degree of responsibility artery stenosis(moderate-severe stenosis[50%-99%stenosis rate],occlusion[100%stenosis rate]),the intracranial collateral circulation status(American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology[ASITN/SIR]collateral circulation grading),cerebrovascular risk factors(hypertension,diabetes,hyperlipidemia,smoking history),and the laboratory test indicators at admission(glycated hemoglobin,triglycerides,total cholesterol,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,blood uric acid,blood homocysteine).QoL was assessed using the stroke impact scale(SIS),covering eight functional domains and a patient-reported overall recovery item.Multimodal MR data were acquired for all subjects.Whole-brain cerebral blood flow(CBF)images were generated using statistics parameter mapping 12(SPM 12)software,while regional homogeneity(ReHo)images were generated using DPABI software.The voxel-wise ratio of CBF to ReHo(CBF/ReHo)was calculated as the regional NVC parameter.Differences in regional NVC characteristics were compared between patient groups and the healthy control group.Correlations between NVC parameters and SIS scores within patient groups were explored.Results(1)A total of 38 patients with MIS or TIA due to intracranial large artery moderate-to-severe stenosis or occlusion were included(26 males,12 females,aged 36-69 years,with mean age of[52±11]years),with 23 in the left-sided involvement group and 15 in the right-sided involvement group.Nineteen healthy subjects were included(10 males,9 females,aged 37-67 years,with mean age of[53±10]years).Post-hoc power analysis showed statistical power of 0.808 for comparing the left-sided involvement group with the healthy control group and 0.762 for comparing the right-sided involvement group with control group.(2)No statistically significant differences were found on gender,age,education level,or body mass index across the three groups(all P＞0.05).No statistically significant differences were observed on the type of cerebrovascular event,cerebrovascular risk factors,distribution of the responsible vessel,degree of stenosis in the responsible vessel,admission NIHSS score,or laboratory test results between the two patient groups(all P＞0.05).There were no statistically significant differences in the total SIS score and the scores of subscales between the two patient groups(all P＞0.05).(3)Compared with the healthy control group,the left-sided involvement group exhibited reduced CBF/ReHo values in the left superior and middle temporal gyri,supramarginal gyrus,middle and inferior frontal gyri,precentral gyrus,angular gyrus,postcentral gyrus,insula,and posterior cerebellar lobe(FDR-corrected,all P＜0.05).In the right-sided involvement group,reduced CBF/ReHo values were observed in the right supramarginal gyrus,right postcentral gyrus,inferior temporal gyrus,and insula(FDR-corrected,all P＜0.05).(4)Correlation analysis revealed that the SIS total score in the left-sided involvement group negatively correlated with CBF/ReHo values in the right inferior frontal gyrus(T=-5.91)and the right middle temporal gyrus(T=-6.65,FDR-corrected,both P＜0.05).The SIS subscale score for activities of daily living in the left-sided involvement group showed negative correlations with CBF/ReHo values in the right angular gyrus(T=-7.36),right medial superior frontal gyrus(T=-6.97),right orbitofrontal cortex(T=-8.99),and left thalamus(T=-7.51,FDR-corrected,all P＜0.05).No significant correlation was observed between the SIS total score and CBF/ReHo values in patients with right-sided involvement group.The SIS subscale for communication score in the right-sided involvement group correlated with CBF/ReHo in the left lingual gyrus(T=-12.15),left olfactory cortex(T=-7.68),and right anterior cingulate and paracingulate cortex(T=-9.46,FDR-corrected,all P＜0.05).Conclusions Patients with MIS or TIA due to intracranial large artery moderate-to-severe stenosis or occlusion show abnormal NVC in the acute phase,especially those with left hemisphere involvement,who exhibit more extensive impairments.QoL in left-sided involvement patients is strongly linked to NVC in the right orbitofrontal cortex and right middle temporal gyrus.These findings require further validation in larger-scale studies.

Objective To investigate the effects of extracellular vesicles(EVs)derived from synovial fluid of rheumatoid arthritis(RA)patients on angiogenesis of human umbilical vein endothelial cells,and to preliminarily explore the underlying mechanisms.Methods Synovial fluid samples of knee joint were collected from 20 patients with RA and 20 patients with osteoarthritis(OA)in this study.EVs were purified using ultracentrifugation.The morphology and size of EVs were observed by transmission electron microscopy and nanoparticle tracking analysis.CD9,CD63,cytochrome c(Cyt-c),vascular endothelial growth factor(VEGF),lysyl oxidase(LOX),matrix metalloproteinase 2(MMP2),tumour necrosis factor alpha(TNF-α),transforming growth factor beta1(TGF-β1)in EVs were detected using Western blot.Human umbilical vein endothelial cells(HUVEC)were treated with the EVs.The growth,migration and angiogenesis of HUVEC were observed by CCK8 assay,TranswellTM chamber assay,scratch test and matrigel angiogenesis assay,respectively.The effect of EVs on the PI3K/AKT pathway in HUVEC was assessed using Western blot.Results Both EVs from RA synovial fluid(RA-EVs)and OA synovial fluid(OA-EVs)were cup-shaped,mainly between 30-400 nm in diameter,expressing CD63 and CD9,but not Cyt-c.RA-EVs carried more VEGF,LOX,MMP2,TNF-α and TGF-β1 than OA-EVs.Compared with the OA-EVs intervention,RA-EVs significantly promoted the proliferation,migration,and angiogenesis of HUVECs,as well as upregulated PI3K/AKT phosphorylation.The inhibitor of PI3K suppressed angiogenesis induced by EVs.Conclusion EVs in synovial fluid of RA carried more cytokines and enzymes that related angiogenesis and inflammation.These EVs exert their pro-angiogenic effects by activating the PI3K/AKT pathway,then contributing to the pathological progression of RA.

Objective To develop an artificial intelligence(AI)diagnostic system for computed tomography angiography(CTA)images related to acute aortic syndrome(AAS)and to evaluate its efficacy in diagnosing and differentiating between AAS subtypes.Methods We collected the CTA images of patients diagnosed with aortic dissection(AD),intramural hematoma(IMH),or penetrating atherosclerotic ulcer(PAU)who were treated in the Department of Peripheral Vascular Diseases,The First Affiliated Hospital of Xi'an Jiaotong University,from June 2016 to June 2022.Based on the strict inclusion and exclusion criteria,2 057 CTA images were selected and extracted.Normal human aorta CTA images served as the control group.The YOLO v7 neural network was used to diagnose and differentiate subgroups of AAS patients in the CTA images,and its diagnostic performance was evaluated.Results The sensitivity,specificity,positive predictive value,negative predictive value,and total accuracy of the AAS diagnostic system based on the YOLO v7 were 98.72％,83.10％,97.82％,89.40％,and 96.92％,respectively.The total accuracy for differential diagnosis of the subgroups was 85.58％.The total accuracy of diagnosis results was significantly higher than that of differential diagnosis results(P＜0.05).Conclusion The AI-based AAS diagnostic system utilizing YOLO v7 meets the established standards for disease diagnosis.However,further research is warranted involving larger image databases and more advanced deep learning networks to improve the differentiation among AAS subtypes.