Objective To investigate the relationship between serum cystatin C(CysC)level and the risk of Parkinson's disease(PD)in middle-aged and elderly people in China.Methods Based on the baseline survey data from the China Health and Retirement Longitudinal Study(CHARLS)in 2011,participants who were not diagnosed with PD at the time of the baseline survey were recruited.The onset of PD was tracked and followed up until 2020,and the participants were divided into PD group and non-PD group according to whether they were newly diagnosed with PD in 2020.Multivariable Logistic regression analysis was performed to assess the association between serum CysC level and the risk of PD.Subgroup and interaction analyses were performed to assess effect modifications by age,gender and depression.Additionally,restricted cubic spline(RCS)was used to explore the linear or non-linear relationship between serum CysC level and the risk of PD in different subgroups.Results We included a total of 3 339 subjects in this study,who consisted of 1 495 males(44.77％)and 1 844 females(55.23％).While baseline participants were followed until 2020,32 subjects had a new PD,and the incidence of PD was 0.96％.The median age of PD group was 63.00 years.Multivariable Logistic regression analysis found that CysC was an independent risk factor for the risk of PD,and CysC was positive significantly associated with the risk of PD(OR=2.34,95％ CI:1.14-4.82,P=0.021).Subgroup analysis showed that CysC was positively associated with PD in females(OR=2.70,95％ CI:1.30-5.58,P=0.007)and subjects aged 60 years or older(OR=5.29,95％ CI:1.69-16.53,P=0.004).RCS model indicated a linear relationship between serum CysC level and the risk of PD in females(Ptotal=0.018,Pnon-linear=0.062)and subjects aged 60 years or older(Ptotal=0.024,Pnon-linear=0.379).Conclusion High level of CysC may increase the risk of PD in middle-aged and elderly people,especially in females and those aged 60 years or older.

Objective:This study explored the effect of nanoparticle-encapsulated curcumin on the highly expressed multidrug resistance gene 1 （MDR1） in a human low-differentiated nasopharyngeal carcinoma cell line （CNE2）. Methods:Curcumin/chitosan deoxycholic acid nanoparticles were prepared, and the cells were subjected to different treatments: radiotherapy, empty carriers, curcumin, and curcumin-loaded nanoparticles. Cell survival was analyzed using the clonogenic assay, and assessments of apoptosis, MDR1 levels, and miR593 levels were conducted. Results:The cell survival fractions in the curcumin group and the curcumin-loaded nanoparticles group were significantly reduced. Notably, higher apoptosis rates were observed in cells treated with curcumin or curcumin-loaded nanoparticles compared to those that received only radiotherapy. Moreover, a decreased MDR1 level was noted in both the curcumin group and the curcumin-loaded nanoparticles group, with further reduction in MDR1 expression observed in the nanoparticle group （P<0.05）. Enhanced expression of miR593 was found in the curcumin group and the curcumin-loaded nanoparticles group, with a relatively higher level in the nanoparticle group （P<0.05）. Curcumin encapsulated in nanoparticles exhibited a stronger radiosensitizing effect. The combination of curcumin and radiotherapy effectively inhibited nasopharyngeal carcinoma （NPC） tumor growth, suppressed MDR1 expression, and enhanced miR593 levels. After inhibiting miR593, MDR1 expression increased. The radiosensitizing effect of curcumin-loaded nanoparticles was regulated by miR593 rather than being triggered by MDR1. Conclusion:Curcumin-loaded nanoparticles mediated enhanced expression of miR593, which in turn inhibited the transcription and translation of the MDR1 gene, thereby reducing the radioresistance of NPC and effectively restraining its growth.
Humans ; Curcumin/pharmacology* ; Nasopharyngeal Neoplasms/pathology* ; Nasopharyngeal Carcinoma ; Nanoparticles ; Cell Line, Tumor ; Apoptosis/drug effects* ; MicroRNAs ; ATP Binding Cassette Transporter, Subfamily B ; ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism* ; Cell Survival

Objective:To establish an automatic classification approach for bone marrow cells based on microscopic hyperspectral imaging and three-dimensional spectral convolutional neural network (Spec-CNN).Methods:The research type is establishment of methodology. The study included 306 newly diagnosed patients' bone marrow smears under Wright's staining from the Department of Hematology of the First Medical Center of the PLA General Hospital from November 1st, 2013 to April 30th, 2024. The high-spectrum data and 4k image data of bone marrow cells were simultaneously collected using a microscopic hyperspectral-4k optical path integrated imaging system (with a spectral resolution of 400—1 000 nm). The high-spectrum data was used for model training, while the 4k image data recognized by morphologists was only used as a reference for labeling the high-spectrum data. The high-spectrum data set was divided into training set, validation set and test set in a ratio of 14∶6∶5. The training set and validation set were used to train and fine-tune the Spec-CNN model, and the test set was used to evaluate the model performance. The sensitivity, specificity ,accuracy ,and Kappa coefficient were calculated for comparing the manual annotation results as gold standard with the intelligent identification results of the Spec-CNN model. Five non-data set samples were used for external validation.Results:The acquired hyperspectral data and 4k imaging dataset comprised of 32 categories and 64 800 bone marrow cells. In the test set, the Spec-CNN model demonstrated weighted-average indicators on classification metrics across 32 cell types: sensitivity 87.79%, specificity 99.31%, and accuracy 98.78%, and Kappa coefficient 0.869. For external validation, the mean correct identification rate of bone marrow cells reached 83.28%.Conclusion:We successfully established an automatic classification method of bone marrow cells based on microscopic hyperspectral imaging and three-dimensional Spec-CNN. This method has a good automatic classification ability for 32 types of bone marrow nucleated cells, which has a certain auxiliary effect on improving the diagnosis efficiency of blood diseases for bone marrow morphologists.

Objective To observe and compare the therapeutic effects of aflibercept biosimilar and originator afliber-cept through intravitreal injection in patients with age-related wet macular degeneration(AMD).Methods This retro-spective study enrolled 60 AMD patients(60 eyes)who were diagnosed and treated at the Department of Ophthalmology,Zibo Central Hospital,from January 2024 to May 2024.These patients were divided into two groups.Specifically,30 pa-tients(30 eyes)(the originator group)received the intravitreal injection of originator aflibercept,while another 30 patients(30 eyes)(the biosimilar group)received the intravitreal injection of aflibercept biosimilar.All patients followed a 3+PRN treatment regimen with a follow-up period of at least 6 months.Relevant examinations including best-corrected visual acui-ty(BCVA),optical coherence tomography,corneal endothelial microscopy,and multifocal electroretinogram(mfERG)were performed before and after treatment.Moreover,the BCVA,central retinal thickness(CRT),pigment epithelial de-tachment(PED)height,corneal endothelial cell density,hexagonal cell ratio,coefficient of variation(CV)of corneal en-dothelial cells,P1 wave amplitude density of the first ring,number of intravitreal injections,and incidence of complications were compared between both groups.Results No statistically significant differences were observed in the age,sex ratio,or disease duration between both groups(all P＞0.05).After treatment,the BCVA,CRT,and PED height of patients in both groups decreased,and the P1 wave amplitude density of the first ring increased in both groups,with statistically sig-nificant differences compared with baseline values(all P＜0.05).In the originator group,the corneal endothelial cell den-sity,hexagonal cell ratio,and CV of corneal endothelial cells of patients in both groups increased after treatment,while the biosimilar group showed a decrease in the corneal endothelial cell density and hexagonal cell ratio,alongside an increase in the CV of corneal endothelial cells;however,these changes were not statistically significant compared with baseline values(all P＞0.05).No significant difference was observed in the number of intravitreal injections between both groups(P＞0.05).The correlation analysis results revealed positive associations between disease duration and post-treatment BCVA(logMAR)in both groups(r=0.901 and 0.905,respectively;both P＜0.000 1).No severe ocular complications or cardio-vascular/cerebrovascular events occurred in all patients.Conclusion There is no significant difference in the short-term efficacy between originator aflibercept and aflibercept biosimilar in the treatment of patients with wet AMD.

Objective:To establish an automatic classification approach for bone marrow cells based on microscopic hyperspectral imaging and three-dimensional spectral convolutional neural network (Spec-CNN).Methods:The research type is establishment of methodology. The study included 306 newly diagnosed patients' bone marrow smears under Wright's staining from the Department of Hematology of the First Medical Center of the PLA General Hospital from November 1st, 2013 to April 30th, 2024. The high-spectrum data and 4k image data of bone marrow cells were simultaneously collected using a microscopic hyperspectral-4k optical path integrated imaging system (with a spectral resolution of 400—1 000 nm). The high-spectrum data was used for model training, while the 4k image data recognized by morphologists was only used as a reference for labeling the high-spectrum data. The high-spectrum data set was divided into training set, validation set and test set in a ratio of 14∶6∶5. The training set and validation set were used to train and fine-tune the Spec-CNN model, and the test set was used to evaluate the model performance. The sensitivity, specificity ,accuracy ,and Kappa coefficient were calculated for comparing the manual annotation results as gold standard with the intelligent identification results of the Spec-CNN model. Five non-data set samples were used for external validation.Results:The acquired hyperspectral data and 4k imaging dataset comprised of 32 categories and 64 800 bone marrow cells. In the test set, the Spec-CNN model demonstrated weighted-average indicators on classification metrics across 32 cell types: sensitivity 87.79%, specificity 99.31%, and accuracy 98.78%, and Kappa coefficient 0.869. For external validation, the mean correct identification rate of bone marrow cells reached 83.28%.Conclusion:We successfully established an automatic classification method of bone marrow cells based on microscopic hyperspectral imaging and three-dimensional Spec-CNN. This method has a good automatic classification ability for 32 types of bone marrow nucleated cells, which has a certain auxiliary effect on improving the diagnosis efficiency of blood diseases for bone marrow morphologists.

Objective To investigate the relationship between serum cystatin C(CysC)level and the risk of Parkinson's disease(PD)in middle-aged and elderly people in China.Methods Based on the baseline survey data from the China Health and Retirement Longitudinal Study(CHARLS)in 2011,participants who were not diagnosed with PD at the time of the baseline survey were recruited.The onset of PD was tracked and followed up until 2020,and the participants were divided into PD group and non-PD group according to whether they were newly diagnosed with PD in 2020.Multivariable Logistic regression analysis was performed to assess the association between serum CysC level and the risk of PD.Subgroup and interaction analyses were performed to assess effect modifications by age,gender and depression.Additionally,restricted cubic spline(RCS)was used to explore the linear or non-linear relationship between serum CysC level and the risk of PD in different subgroups.Results We included a total of 3 339 subjects in this study,who consisted of 1 495 males(44.77％)and 1 844 females(55.23％).While baseline participants were followed until 2020,32 subjects had a new PD,and the incidence of PD was 0.96％.The median age of PD group was 63.00 years.Multivariable Logistic regression analysis found that CysC was an independent risk factor for the risk of PD,and CysC was positive significantly associated with the risk of PD(OR=2.34,95％ CI:1.14-4.82,P=0.021).Subgroup analysis showed that CysC was positively associated with PD in females(OR=2.70,95％ CI:1.30-5.58,P=0.007)and subjects aged 60 years or older(OR=5.29,95％ CI:1.69-16.53,P=0.004).RCS model indicated a linear relationship between serum CysC level and the risk of PD in females(Ptotal=0.018,Pnon-linear=0.062)and subjects aged 60 years or older(Ptotal=0.024,Pnon-linear=0.379).Conclusion High level of CysC may increase the risk of PD in middle-aged and elderly people,especially in females and those aged 60 years or older.

Objective To observe and compare the therapeutic effects of aflibercept biosimilar and originator afliber-cept through intravitreal injection in patients with age-related wet macular degeneration(AMD).Methods This retro-spective study enrolled 60 AMD patients(60 eyes)who were diagnosed and treated at the Department of Ophthalmology,Zibo Central Hospital,from January 2024 to May 2024.These patients were divided into two groups.Specifically,30 pa-tients(30 eyes)(the originator group)received the intravitreal injection of originator aflibercept,while another 30 patients(30 eyes)(the biosimilar group)received the intravitreal injection of aflibercept biosimilar.All patients followed a 3+PRN treatment regimen with a follow-up period of at least 6 months.Relevant examinations including best-corrected visual acui-ty(BCVA),optical coherence tomography,corneal endothelial microscopy,and multifocal electroretinogram(mfERG)were performed before and after treatment.Moreover,the BCVA,central retinal thickness(CRT),pigment epithelial de-tachment(PED)height,corneal endothelial cell density,hexagonal cell ratio,coefficient of variation(CV)of corneal en-dothelial cells,P1 wave amplitude density of the first ring,number of intravitreal injections,and incidence of complications were compared between both groups.Results No statistically significant differences were observed in the age,sex ratio,or disease duration between both groups(all P＞0.05).After treatment,the BCVA,CRT,and PED height of patients in both groups decreased,and the P1 wave amplitude density of the first ring increased in both groups,with statistically sig-nificant differences compared with baseline values(all P＜0.05).In the originator group,the corneal endothelial cell den-sity,hexagonal cell ratio,and CV of corneal endothelial cells of patients in both groups increased after treatment,while the biosimilar group showed a decrease in the corneal endothelial cell density and hexagonal cell ratio,alongside an increase in the CV of corneal endothelial cells;however,these changes were not statistically significant compared with baseline values(all P＞0.05).No significant difference was observed in the number of intravitreal injections between both groups(P＞0.05).The correlation analysis results revealed positive associations between disease duration and post-treatment BCVA(logMAR)in both groups(r=0.901 and 0.905,respectively;both P＜0.000 1).No severe ocular complications or cardio-vascular/cerebrovascular events occurred in all patients.Conclusion There is no significant difference in the short-term efficacy between originator aflibercept and aflibercept biosimilar in the treatment of patients with wet AMD.

Objective:To investigate the expression, distribution and cellular localization of acylglycerol kinase (AGK) in liver cancer tissues, and the correlation of AGK expression with the prognosis of liver cancer patients.Methods:AGK mRNA expression data and clinical information of hepatocellular carcinoma (HCC) patients were downloaded from The Cancer Genome Atlas (TCGA) database in January 2024. The expression differences of AGK mRNA between HCC tissues and paracancerous tissues were compared, and the high and low expressions of AGK were judged by using the median expression of AGK mRNA in 369 HCC tissues as a cut-off value. A univariate logistic regression model was used to analyze the relationship between clinical pathological characteristics and high expression of AGK. The mRNA expressions in HCC tissues and paracancerous tissues of 9 datasets from the Hepatocellular Carcinoma Molecular Landscape Database (HCCDB) 2.0 were compared. The spatial distribution and cellular localization of AGK were analyzed based on multidimensional data from Bulk transcriptome sequencing (RNA-seq), single-cell sequencing and spatial RNA-seq. The expression of AGK protein in liver cancer tissues was analyzed using the Human Protein Atlas (HPA) database. Kaplan-Meier method and log-rank test were employed to compare the differences in overall survival (OS) among patients with different AGK mRNA expressions in HCCDB25 dataset of HCCDB 2.0 and HPA database. The correlation between expressions of AGK and hepatic stem cell-related markers was analyzed by using Spearman rank test based on Tumor Immune Estimation Resource (TIMER) 2.0 database.Results:Data from both the TCGA and 9 datasets of HCCDB 2.0 showed that AGK mRNA expression in HCC tissues was higher than that in paracancerous non-tumorous tissues and normal liver tissues, and the difference was statistically significant (all P < 0.001). HPA database immunohistochemical testing revealed that AGK protein was primarily localized in the cytoplasm, with positive or strong positive expression in HCC tissues and negative or weak positive expression in normal liver tissues; mass spectrometry data showed that it was upregulated in tumor samples (165 cases) compared to normal liver tissues (165 cases) ( P < 0.001). Univariate logistic regression analysis indicated that tumor family history and tumor pathological differentiation in HCC patients from TCGA database were associated with high AGK expression in tumor tissues ( P values were 0.028 and 0.050), while other factors such as age, gender, body mass index, alpha fetoprotein level, Child-Pugh classification, inflammation degree in paracancerous tissues, Ishak fibrosis score, pathological TNM staging, tumor clinical staging, and tumor vascular infiltration had no impact on AGK expression level in tumor tissues (all P > 0.05). One hundred and fifty-eight patients were divided into high and low KGK mRNA expression groups based on the median expression of AGK mRNA in tissues of HCCDB25 dataset, analysis showed that patients in low AGK mRNA expression group (79 cases) had better overall survival (OS) compared to the high expression group (79 cases) in tumor tissues, and the difference was statistically significant ( P = 0.038), while there was no significant difference in OS between high (79 cases) and low (79 cases) expression groups in paracancerous tissues ( P = 0.760). In HPA database, patients were divided into high and low AGK mRNA expression groups based on AGK mRNA values in liver cancer tissues corresponding to the lowest P value during OS analysis by Kaplan-Meier method; in all stages of HCC patients, low AGK mRNA expression group (279 cases) had better OS than the high expression group (76 cases), and the difference was statistically significant ( P = 0.022). The OS of high AGK mRNA expression group in patients with stages Ⅱ-Ⅲ was worse than that of low expression group, and the difference was statistically significant ( P = 0.007). The UMAP plot obtained through dimensionality reduction and cell clustering analysis based on single-cell sequencing data in HCCDB 2.0 revealed that AGK gene expression in liver cancer tissues was primarily distributed in tumor cells, NK/T cells, stromal cells, and myeloid cells. Spatial transcriptomic analysis of tissue samples from 5 HCC patients using HCCDB 2.0 online tools showed that AGK expression varied across different liver cancer tissue regions (non-tumorous tissue, paracancerous tissue, tumor junction, tumor focus, and portal vein tumor thrombus), with 3 cases showing AGK expression enrichment in tumor cells of the tumor junction, tumor focus and portal vein tumor thrombus, while lower in normal hepatocytes, stromal cells and immune cells. In 2 cases, AGK expression was more widespread. Analysis of 3 patients with significant AGK enrichment showed that in HCC samples with complete fibrous capsule, AGK was mainly localized in tumor cells of the tumor focus and junction areas, with weaker expression in paracancerous normal tissues; while in samples with incomplete capsule, high AGK expression was primarily in tumor cells of the tumor junction, tumor focus and portal vein tumor thrombus. TIMER 2.0 database assessment showed that AGK gene expression in 371 patients of TCGA database was positively correlated with the expressions of liver cancer stem cell-related marker genes, including PROM1 ( rho = 0.250), TYH1 ( rho = 0.188), CD44 ( rho = 0.268), ANPEP ( rho = 0.171), CD47 ( rho = 0.435), EPCAM ( rho = 0.246), KRT19 ( rho = 0.203), TGFB1 ( rho = 0.285), and SOX9 ( rho = 0.328) (all P < 0.001). Conclusions:AGK expression is significantly upregulated at both mRNA and protein levels in tumor tissues of HCC patients, it predominantly localizes in the tumor tissues and the cytoplasm of tumor cells within the junction areas, and its high expression closely associates with poor prognosis of patients. Its expression is positively correlated with the expression of liver cancer stem cell-related markers.

Chemotherapy resistance plays a pivotal role in the prognosis and therapeutic failure of patients with colorectal cancer(CRC).Cisplatin(DDP)-resistant cells exhibit an inherent ability to evade the toxic chemotherapeutic drug effects which are characterized by the activation of slow-cycle programs and DNA repair.Among the elements that lead to DDP resistance,O6-methylguanine(O6-MG)-DNA-meth-yltransferase(MGMT),a DNA-repair enzyme,performs a quintessential role.In this study,we clarify the significant involvement of MGMT in conferring DDP resistance in CRC,elucidating the underlying mechanism of the regulatory actions of MGMT.A notable upregulation of MGMT in DDP-resistant cancer cells was found in our study,and MGMT repression amplifies the sensitivity of these cells to DDP treatment in vitro and in vivo.Conversely,in cancer cells,MGMT overexpression abolishes their sensi-tivity to DDP treatment.Mechanistically,the interaction between MGMT and cyclin dependent kinase 1(CDK1)inducing slow-cycling cells is attainted via the promotion of ubiquitination degradation of CDK1.Meanwhile,to achieve nonhomologous end joining,MGMT interacts with XRCC6 to resist chemotherapy drugs.Our transcriptome data from samples of 88 patients with CRC suggest that MGMT expression is co-related with the Wnt signaling pathway activation,and several Wnt inhibitors can repress drug-resistant cells.In summary,our results point out that MGMT is a potential therapeutic target and predictive marker of chemoresistance in CRC.

Objective:To investigate the expression, distribution and cellular localization of acylglycerol kinase (AGK) in liver cancer tissues, and the correlation of AGK expression with the prognosis of liver cancer patients.Methods:AGK mRNA expression data and clinical information of hepatocellular carcinoma (HCC) patients were downloaded from The Cancer Genome Atlas (TCGA) database in January 2024. The expression differences of AGK mRNA between HCC tissues and paracancerous tissues were compared, and the high and low expressions of AGK were judged by using the median expression of AGK mRNA in 369 HCC tissues as a cut-off value. A univariate logistic regression model was used to analyze the relationship between clinical pathological characteristics and high expression of AGK. The mRNA expressions in HCC tissues and paracancerous tissues of 9 datasets from the Hepatocellular Carcinoma Molecular Landscape Database (HCCDB) 2.0 were compared. The spatial distribution and cellular localization of AGK were analyzed based on multidimensional data from Bulk transcriptome sequencing (RNA-seq), single-cell sequencing and spatial RNA-seq. The expression of AGK protein in liver cancer tissues was analyzed using the Human Protein Atlas (HPA) database. Kaplan-Meier method and log-rank test were employed to compare the differences in overall survival (OS) among patients with different AGK mRNA expressions in HCCDB25 dataset of HCCDB 2.0 and HPA database. The correlation between expressions of AGK and hepatic stem cell-related markers was analyzed by using Spearman rank test based on Tumor Immune Estimation Resource (TIMER) 2.0 database.Results:Data from both the TCGA and 9 datasets of HCCDB 2.0 showed that AGK mRNA expression in HCC tissues was higher than that in paracancerous non-tumorous tissues and normal liver tissues, and the difference was statistically significant (all P < 0.001). HPA database immunohistochemical testing revealed that AGK protein was primarily localized in the cytoplasm, with positive or strong positive expression in HCC tissues and negative or weak positive expression in normal liver tissues; mass spectrometry data showed that it was upregulated in tumor samples (165 cases) compared to normal liver tissues (165 cases) ( P < 0.001). Univariate logistic regression analysis indicated that tumor family history and tumor pathological differentiation in HCC patients from TCGA database were associated with high AGK expression in tumor tissues ( P values were 0.028 and 0.050), while other factors such as age, gender, body mass index, alpha fetoprotein level, Child-Pugh classification, inflammation degree in paracancerous tissues, Ishak fibrosis score, pathological TNM staging, tumor clinical staging, and tumor vascular infiltration had no impact on AGK expression level in tumor tissues (all P > 0.05). One hundred and fifty-eight patients were divided into high and low KGK mRNA expression groups based on the median expression of AGK mRNA in tissues of HCCDB25 dataset, analysis showed that patients in low AGK mRNA expression group (79 cases) had better overall survival (OS) compared to the high expression group (79 cases) in tumor tissues, and the difference was statistically significant ( P = 0.038), while there was no significant difference in OS between high (79 cases) and low (79 cases) expression groups in paracancerous tissues ( P = 0.760). In HPA database, patients were divided into high and low AGK mRNA expression groups based on AGK mRNA values in liver cancer tissues corresponding to the lowest P value during OS analysis by Kaplan-Meier method; in all stages of HCC patients, low AGK mRNA expression group (279 cases) had better OS than the high expression group (76 cases), and the difference was statistically significant ( P = 0.022). The OS of high AGK mRNA expression group in patients with stages Ⅱ-Ⅲ was worse than that of low expression group, and the difference was statistically significant ( P = 0.007). The UMAP plot obtained through dimensionality reduction and cell clustering analysis based on single-cell sequencing data in HCCDB 2.0 revealed that AGK gene expression in liver cancer tissues was primarily distributed in tumor cells, NK/T cells, stromal cells, and myeloid cells. Spatial transcriptomic analysis of tissue samples from 5 HCC patients using HCCDB 2.0 online tools showed that AGK expression varied across different liver cancer tissue regions (non-tumorous tissue, paracancerous tissue, tumor junction, tumor focus, and portal vein tumor thrombus), with 3 cases showing AGK expression enrichment in tumor cells of the tumor junction, tumor focus and portal vein tumor thrombus, while lower in normal hepatocytes, stromal cells and immune cells. In 2 cases, AGK expression was more widespread. Analysis of 3 patients with significant AGK enrichment showed that in HCC samples with complete fibrous capsule, AGK was mainly localized in tumor cells of the tumor focus and junction areas, with weaker expression in paracancerous normal tissues; while in samples with incomplete capsule, high AGK expression was primarily in tumor cells of the tumor junction, tumor focus and portal vein tumor thrombus. TIMER 2.0 database assessment showed that AGK gene expression in 371 patients of TCGA database was positively correlated with the expressions of liver cancer stem cell-related marker genes, including PROM1 ( rho = 0.250), TYH1 ( rho = 0.188), CD44 ( rho = 0.268), ANPEP ( rho = 0.171), CD47 ( rho = 0.435), EPCAM ( rho = 0.246), KRT19 ( rho = 0.203), TGFB1 ( rho = 0.285), and SOX9 ( rho = 0.328) (all P < 0.001). Conclusions:AGK expression is significantly upregulated at both mRNA and protein levels in tumor tissues of HCC patients, it predominantly localizes in the tumor tissues and the cytoplasm of tumor cells within the junction areas, and its high expression closely associates with poor prognosis of patients. Its expression is positively correlated with the expression of liver cancer stem cell-related markers.