Reprogramming tumor microenvironment with precise photothermal therapy by calreticulin nanobody-engineered probiotics.

Targeted therapies have revolutionized traditional cancer treatments by precisely targeting tumor cells, enhancing efficacy and safety. Despite this advancement, the proportion of cancer patients eligible for such therapies remains low due to the absence of suitable targets. Here, we investigate whether the translocation of the immunogenic cell death (ICD) marker calreticulin (CALR) from the endoplasmic reticulum (ER) to the cell surface following ICD induction can serve as a target for targeted therapies. To target CALR, a nanobody Nb215 identified from a naïve VHH phage library with high binding affinity to both human and mouse CALR was employed to engineer probiotic EcN 1917. Our results demonstrated that CALR nanobody-modified EcN-215 coupled with the photothermal dye indocyanine green (ICG) was able to exert NIR-II imaging-guide photothermal therapy (PTT). Moreover, PTT with EcN-215/ICG can reshape the tumor microenvironment by enhancing the infiltration of CD45+CD3+ T cells and CD11b+F4/80+ macrophages. Furthermore, the antitumor activity of CALR-targeted EcN-215/ICG is synergistically enhanced by blocking CD47-SIRPα axis. Collectively, our study provides a proof of concept for CALR-targeted therapy. Given that CALR translocation can be induced by various anticancer therapies across numerous tumor cell lines, CALR-targeted therapies hold promise as a novel approach for treating multiple types of cancers.

Off-the-shelf CAR-NK cells targeting immunogenic cell death marker ERp57 execute robust antitumor activity and have a synergistic effect with ICD inducer oxaliplatin.

BACKGROUND: Chimeric antigen receptor natural killer (CAR-NK) therapy holds great promise for treating hematologic tumors, but its efficacy in solid tumors is limited owing to the lack of suitable targets and poor infiltration of engineered NK cells. Here, we explore whether immunogenic cell death (ICD) marker ERp57 translocated from endoplasmic reticulum to cell surface after drug treatment could be used as a target for CAR-NK therapy. METHODS: To target ERp57, a VHH phage display library was used for screening ERp57-targeted nanobodies (Nbs). A candidate Nb with high binding affinity to both human and mouse ERp57 was used for constructing CAR-NK cells. Various in vitro and in vivo studies were performed to assess the antitumor efficacy of the constructed CAR-NK cells. RESULTS: We demonstrate that the translocation of ERp57 can not only be induced by low-dose oxaliplatin (OXP) treatment but also is spontaneously expressed on the surface of various types of tumor cell lines. Our results show that G6-CAR-NK92 cells can effectively kill various tumor cell lines in vitro on which ERp57 is induced or intrinsically expressed, and also exhibit potent antitumor effects in cancer cell-derived xenograft and patient-derived xenograft mouse models. Additionally, the antitumor activity of G6-CAR-NK92 cells is synergistically enhanced by the low-dose ICD-inducible drug OXP. CONCLUSION: Collectively, our findings suggest that ERp57 can be leveraged as a new tumor antigen for CAR-NK targeting, and the resultant CAR-NK cells have the potential to be applied as a broad-spectrum immune cell therapy for various cancers by combining with ICD inducer drugs.

Cadherin 17 Nanobody-Mediated Near-Infrared-II Fluorescence Imaging-Guided Surgery and Immunotoxin Delivery for Colorectal Cancer.

Surgery and targeted therapy are of equal importance for colorectal cancer (CRC) treatment. However, complete CRC tumor resection remains challenging, and new targeted agents are also needed for efficient CRC treatment. Cadherin 17 (CDH17) is a membrane protein that is highly expressed in CRC and, therefore, is an ideal target for imaging-guided surgery and therapeutics. This study utilizes CDH17 nanobody (E8-Nb) with the near-infrared (NIR) fluorescent dye IRDye800CW to construct a NIR-II fluorescent probe, E8-Nb-IR800CW, and a Pseudomonas exotoxin (PE)-based immunotoxin, E8-Nb-PE38, to evaluate their performance for CRC imaging, imaging-guided precise tumor excision, and antitumor effects. Our results show that E8-Nb-IR800CW efficiently recognizes CDH17 in CRC cells and tumor tissues, produces high-quality NIR-II images for CRC tumors, and enables precise tumor removal guided by NIR-II imaging. Additionally, fluorescent imaging confirms the targeting ability and specificity of the immunotoxin toward CDH17-positive tumors, providing the direct visible evidence for immunotoxin therapy. E8-Nb-PE38 immunotoxin markedly delays the growth of CRC through the induction of apoptosis and immunogenic cell death (ICD) in multiple CRC tumor models. Furthermore, E8-Nb-PE38 combined with 5-FU exerts synergistically antitumor effects and extends survival. This study highlights CDH17 as a promising target for CRC imaging, imaging-guided surgery, and drug delivery. Nanobodies targeting CDH17 hold great potential to construct NIR-II fluorescent probes for surgery navigation, and PE-based toxins fused with CDH17 nanobodies represent a novel therapeutic strategy for CRC treatment. Further investigation is warranted to validate these findings for potential clinical translation.

Validation of the Modified Location-based Resect-and-discard Strategy Requiring Pathology Examination of Sigmoid Diminutive Polyps.

BACKGROUND AND AIMS: Recently, the location-based resect-and-discard (LBRD) strategy, which does not depend on optical diagnosis, was developed and demonstrated different surveillance interval agreement with the pathology-based reference in several researches. We aimed to evaluate the performance of LBRD in our first-time colonoscopy cohort, and improve the LBRD. METHODS: The first-time colonoscopy with complete pathologic information were enrolled. The accuracy of LBRD strategy applied in diminutive polyps in different colonic segments was used as indicator to develop modified LBRD (mLBRD) strategy. Surveillance interval agreement with pathology-based reference was compared between LBRD and mLBRD. The ≥ 90% agreement with pathology was used as benchmark. RESULTS: The polyps in sigmoid colon were significantly associated with higher proportion of neoplastic compared with polyps in rectum. The accuracy of LBRD applied in polyps in sigmoid colon were only 53.5%, which was significantly lower than that applied in polyps in other colonic segments. Thus, we hypothesized that mLBRD requiring pathology examination of diminutive polyps in sigmoid colon was more efficient in clinical use. The mLBRD significantly outperformed LBRD in surveillance interval agreement with pathology-based reference (90.2% vs. 83.4%, P<0.001), had lower proportion of patients assigned a longer surveillance interval (3.6% vs. 10.5%, P<0.001) and reached the benchmark, although the proportion of patients with an immediate surveillance interval recommendations and pathology examination avoided decreased. CONCLUSIONS: The mLBRD, but not LBRD, achieved sufficient surveillance interval agreement with pathology-based surveillance interval assignment and reduced over 30% of pathology examinations.

Long-distance migration law of radon in overburden of abandoned goaf during coal spontaneous combustion.

The surface isotope radon measurement method (SIRMM) is widely used in fire source detection in abandoned mines. However, studies on the long-distance migration of radon during coal spontaneous combustion are lacking, which hinders the further popularization of this technology in coal fire prevention and control. For this reason, the migration law of radon in overlying strata in fire areas was studied through experiments and numerical simulation. The radon exhalation concentration of coal was found to increase at first and then decrease in the range of 30-350 °C through experiments. The radon concentration reaches the maximum value (557.1 Bq/m3) at 150 °C, which is 6.3 times higher than that at 30 °C. Based on the radon source term obtained by fitting the experimental data, the radon migration model of coal spontaneous combustion in abandoned goaf was constructed, and the dynamic distribution characteristics of the airflow, temperature, and radon concentration fields in the overlying strata area were analyzed. The internal relationship between surface radon and underground fire source was discussed. The simulation results revealed the sharp change in the porosity of the overlying rock causes radon concentration at the interface between the caving and fissure zones to increase continually with the process of spontaneous combustion, providing material and energy support for the long-distance radon migration. When the maximum temperature of the coal pile reaches 70 °C, the concentration of radon released from the coal pile increases rapidly to 13696 Bq/m3, and the radon from the underground space appears on the surface at this temperature. In the range of 70-150 °C, with rapid increase in radon released from coal piles, the surface concentration of radon also increased rapidly to 225 Bq/m3. At the high-temperature stage exceeding 150 °C, the concentration of radon released from coal piles exhibited a downward trend, resulting in a decrease in the rate of increase of radon concentration on the surface. A close relationship between the surface radon concentration and underground fire source temperature in the process of coal spontaneous combustion was observed. In the spatial position, the peak position of radon on the surface was highly consistent with that of the fire source longitudinally, which ensures the accuracy of the SIRMM to determine the location of the hidden fire source. This suggests that the SIRMM can accurately evaluate the fire source's temperature and fire area's development trend.

Wet Catalytic Oxidation of a FeMnCe-Activated Semi-Coke Catalyst for Treating Piperazine Wastewater.

A FeMnCe-activated semi-coke catalyst (FeMnCe/ASC) was prepared by the co-precipitation method using semi-coke as the raw material. The structure and morphology were characterized by X-ray diffraction, Brunauer-Emmett-Teller, scanning electron microscopy, and transmission electron microscopy analyses. The catalytic activity and stability of the FeMnCe/ASC catalyst were investigated with piperazine as the target degradation pollutant and ammonia nitrogen and chemical oxygen demand (COD) as the evaluation indexes. The results showed that the average pore size of FeMnCe/ASC mesopores was 6.68 nm, and the active components were uniformly dispersed on the carrier surface. Under the optimum conditions of piperazine solution including a mass concentration of 100 mg/L, a catalyst mass concentration of 4.0 g/L, a reaction temperature of 240 °C, an oxygen partial pressure of 1.2 MPa, a stirring speed of 500 rpm, and a reaction time of 120 min, the degradation rates of both ammonia nitrogen and COD reached 100%. After the catalyst was recycled five times, the degradation rates of ammonia nitrogen and COD still reached more than 90%. The elemental valence changes before and after the reaction were analyzed by X-ray photoelectron spectroscopy, and the intermediate products generated from piperazine degradation were analyzed by gas chromatography-mass spectroscopy to evaluate the mechanism of piperazine degradation and speculate about the degradation pathway of piperazine.

Upregulation of Netrin-1 in the hippocampus mediates the formation of visceral hypersensitivity induced by maternal separation.

Early adverse life events (EALs), such as maternal separation (MS), can cause visceral hypersensitivity, which is thought to be a key pathophysiological mechanism of irritable bowel syndrome (IBS). Previous studies mainly focused on EALs-induced visceral hypersensitivity in adulthood but did not consider that it may have occurred in the preadult period. We previously found that rats who experienced MS suffered from visceral hypersensitivity starting from the post-weaning period. Moreover, the hippocampus is considered to be critical in regulating the formation of visceral hypersensitivity induced by MS. But the underlying mechanisms throughout different life periods are unclear. In this study, behavioral tests, RNA-seq, lentiviral interference, and molecular biology techniques were applied to investigate the molecular mechanism in the hippocampus underlying MS-induced long-lasting visceral hypersensitivity. It was found that both visceral sensitivity and anxiety-like behaviors were significantly increased in MS rats in post-weaning, prepubertal, and adult periods, especially in the prepubertal period. Subsequently, RNA-seq targeting the hippocampus identified that the expression level of Netrin-1 was significantly increased in all periods, which was further confirmed by quantitative real-time PCR and Western blot. Knocking-down hippocampal Netrin-1 in the post-weaning period by lentivirus interference alleviated visceral hypersensitivity and anxiety-like behaviors of MS rats in the later phase of life. In addition, deleted in colorectal cancer (DCC), instead of neogenin-1(Neo-1) or uncoordinated (UNC5), was proved to be the specific functional receptor of Netrin-1 in regulating visceral hypersensitivity, whose upregulation may result in the most severe symptoms in the prepubertal period. Furthermore, the activation of the Netrin-1/DCC pathway could enhance long-term potentiation (LTP) in the hippocampus, probably via recruitment of the AMPA receptor subunit GluA1, which finally resulted in the formation of visceral hypersensitivity. These novel findings suggest that long-lasting over-expression of Netrin-1 can mediate visceral hypersensitivity and anxiety disorder from the post-weaning period to adulthood by activating DCC/GluA1 pathway in the hippocampus. Moreover, early intervention of Netrin-1 in the post-weaning period could lead to significant symptom relief afterward, which provides evidence that the Netrin-1/DCC/GluA1 signaling pathway may be a potential therapeutic target for the treatment of visceral hypersensitivity in clinics.

Discrepancy Between Forceps Biopsy and Resection in Colorectal Polyps: A 1686 Paired Screening-Therapeutic Colonoscopic Finding.

Purpose: To identify pathology discrepancy between forceps biopsies and polypectomy specimens in colorectal polyps, as well as the reliability of biopsy-based treatment strategy. Methods: All endoscopic polypectomy cases with forceps biopsies performed within 6 months were included in the study. The biopsies were compared with polypectomy specimens in terms of concordance of histological diagnosis. A logistic regression model was used to investigate the independent predictors of upgrade in histological diagnosis compared with concordance in histological diagnosis. Results: A total of 1686 paired screening-therapeutic colonoscopies and 1739 paired biopsy-polypectomy specimens were enrolled in the study. The grade of dysplasia in 84.5% of biopsy specimens were concordant to polypectomy specimens, but this proportion decreased to 75.4% when the specimens were classified using tubular or villousness structure. 10.1% and 5.4% of biopsy specimens were upgraded and downgraded in assessing grade of dysplasia, respectively, while 14.3% and 10.3% of biopsy specimens were upgraded and downgraded in assessing tubular or villousness structure, respectively. In subgroup analysis stratified by size of polyps, 9.0% and 10.6% of biopsies obtained from polyps smaller than 10 mm were upgraded in assessing dysplasia and tubular or villousness structure, respectively. This proportion increased to 10.7% and 21.3%, respectively, in biopsies obtained from polyps larger than 10 mm. Larger size of polyps and pedunculated polyps were associated with a higher incidence of upgrade in histological diagnosis. Nearly 25% of biopsy specimens with high-grade dysplasia were identified as adenocarcinoma in polypectomy specimens. Conclusion: The concordance between biopsy and polypectomy specimens is not adequate. The biopsy-based treatment strategy is not reliable and should not be considered as an indicator for further treatment, particularly in large or pedunculated polyps.

Negative Effects of Endoscopists' Fatigue on Colonoscopy Quality on 34,022 Screening Colonoscopies.

BACKGROUND AND AIMS: There is still considerable controversy surrounding the relationship between fatigue of endoscopists and the quality of colonoscopy. The aim of this study is to comprehensively explore the association between fatigue and adenoma detection rate (ADR) and cecal intubation rate (CIR). METHODS: The mixed effects logistic regression model was used to explore the relationship between fatigue- related factors including procedure order, session of procedures and the day of week and ADR as well as CIR. RESULTS: When controlling for confounders, the day of week (Monday as reference, Friday, p=0.022; weekends, p=0.015) and session of procedures (P<0.001) were significantly associated with ADR while procedure order (<5 as reference, 6-10, p<0.001; >10, p=0.001) and session of procedures (p=0.004) were independent predictors for CIR. Additionally, there was a significant downward trend on ADR and CIR with the approaching of weekends (p=0.005) and increasing procedure orders (p<0.001), respectively. In the subgroup analysis stratified by gender, age and workload intensity, significant lower ADR was found in the afternoon in all subgroups (male, p<0.001; female, p=0.005; <40 years, p<0.001; ≥40 years, p=0.020; intensity<50 per month, p=0.017; intensity≥50 per month, p<0.001) but the downward trend on ADR as the week progressed was only found in endoscopists with male gender (p=0.011), age<40 (p=0.027) and high workload intensity (p=0.003). Moreover, a significant downward trend on CIR as the procedure order increased was found in all subgroups except endoscopists with age≥40 (male, p=0.005; female, p<0.001; <40 years, p<0.001; intensity<50 per month, p=0.001; intensity≥50 per month, p<0.001). CONCLUSIONS: Colonoscopies in the afternoon will affect ADR negatively while increasing procedure order will cause a lower CIR. Importantly, the significant negative influence of Friday and weekends on ADR was first discovered in this study. Moreover, endoscopists with female gender and advanced age (≥40) but not high workload intensity showed superiority in resistance of fatigue caused by the end of the week and increasing daily procedures.

Comprehensive evaluation of the learning curve to achieve satisfactory adenoma detection rate.

BACKGROUND AND AIM: The number of colonoscopies required to reach satisfactory adenoma detection rate (ADR) is not well established. The aim of this study was to identify the appropriate number of procedures required to attain satisfactory ADR for those well-trained endoscopists who have a cecal intubation rate (CIR) ≥ 90% and start to perform colonoscopy independently. METHODS: All endoscopists with compelete independent colonoscopy data during career in our database were enrolled. The number of procedures required to achieve ADR ≥ 20% was identified by cumulative summation (Cusum), learning curve Cusum (LC-Cusum), and moving average method. Mixed effect logistic regression model was developed to determine the relationship between endoscopist as well as patient-related factors and adenoma detection. RESULTS: A total of 24 943 procedures and 14 endoscopists were enrolled. By Cusum analysis, the interest point was at 207 procedures. By LC-Cusum analysis, 71% (10/14) and 86% (12/14) of endoscopists had attained satisfactory ADR after 200 and 300 procedures, respectively. By moving average method, endoscopists reached a mean ADR of 20% at 216 and 261 procedures over blocks of 50 and 100 procedures, respectively. The total number of procedures, number of daily procedures, patient age and gender, bowel preparation, sedation, and diverticulosis were significantly associated with adenoma detection. CONCLUSIONS: This is the first study to investigate the learning curve of ADR for those well-trained endoscopists who have a CIR ≥ 90% and start to perform colonoscopy independently. Two hundred procedures might be an optimal number required to reach an ADR ≥ 20%.

The Impact of Sedation on Adenoma Detection Rate and Cecal Intubation Rate in Colonoscopy.

PURPOSE: To explore the effect of sedation on the quality of colonoscopy. METHODS: The data collected from the Digestive Endoscopy Center of Shanghai Tongji Hospital from March 2012 to June 2019 were retrospectively analyzed. The rate of sedation and quality metrics of colonoscopy such as adenoma detection rate (ADR) and cecal intubation rate (CIR) were calculated. The logistic regression model was used to explore the relationship between sedation and quality metrics of colonoscopy. The interaction effects between experience of endoscopists and sedation on quality of colonoscopy was also investigated in subgroups stratified by total number of colonoscopies during career using the logistic regression model. RESULTS: A total of 63,417 colonoscopies including 11,417 colonoscopies without sedation and 52,000 colonoscopies with sedation were enrolled in our study. The proportion of colonoscopy with sedation was 82.0%. The ADR and CIR were all significantly higher in cases with sedation compared with cases without sedation (ADR, 22.5% vs. 17.0%, p < 0.001; CIR, 94.7% vs. 91.2%, p < 0.001). Multivariate analysis showed that the sedation was an independent factor associated with adenoma detection (OR = 1.448, 95% CI: 1.372~1.529, p < 0.001) and cecal intubation (OR = 1.560, 95% CI: 1.446~1.683, p < 0.001). A total of 14 endoscopists with complete colonoscopy data in our database and corresponding 20,949 colonoscopies data were enrolled for further analysis. The logistic regression model yielded a similar result that sedation was an independent factor on adenoma detection and cecal intubation when the factor, experience of endoscopists, was also entered into the model as a confounder (adenoma detection, OR = 1.408, 95% CI: 1.333~1.487, p < 0.001; cecal intubation, OR = 1.601, 95% CI: 1.482-1.729, p < 0.001). CONCLUSION: Colonoscopy with sedation has a positive effect on ADR and CIR in all endoscopists with different experience of colonoscopy, which makes the quality of colonoscopy better.

Dichondra repens J.R.Forst. and G.Forst.: A Review of Its Traditional Uses, Chemistry, Pharmacology, Toxicology and Applications.

Ethnopharmacology relevance: Dichondra repens J.R.Forst. and G.Forst (DRF; Convolvulaceae, called Matijin in Chinese), has been traditionally used to treat jaundice, bacillary dysentery, urinary tract infection, edema, contusions, and strains and sprains based on traditional Chinese medicine (TCM) concepts. Aim of study: This paper intends to provide a comprehensive and critical analysis of research on DRF focusing on a relationship between traditional uses and pharmacological effects, evaluating the therapeutic potential of this plant. Methods: Relevant data on DRF were retrieved from available databases. Results: The heat-clearing and detoxifying, and removing the phlegm and turbid urine effects of DRF are linked to its anti-hepatitis B virus (HBV), anti-inflammatory, and hepatoprotective activities. Especially, the hepatoprotective effects of DRF are mainly based on anti-HBV activities of phenylalanine dipeptides Matijin-Su (MTS) and its derivatives derived from this plant. Further, a phase I anti-HBV clinical trial of a candidate compound named bentysrepinine (Y101, Chinese name Tifentai) has been completed. Also, anti-tumor, analgesic, and antibacterial properties have been reported in the extracts and compounds from DRF. Although pharmacy, pharmacodynamics, toxicology, and pharmacokinetics of bentysrepinine have been systemically reported, no studies have reported chemistry, safety, pharmacology of other compounds or extracts systemically. Conclusion: Phenylalanine dipeptide compounds are main components and MTS is a characteristic substance of DRF. The main pharmacological effect of DRF is anti-HBV activity, which is coherent with the traditional use of this plant in China. Except bentysrepinine, few studies have been conducted on toxicities of the extracts or compounds from DRF. Thus, it is still necessary to evaluate safety, chemistry, pharmacology of the extracts or compounds from DRF regarding the link between traditional uses and modern applications before the future clinical trials. Bacterial sepsis, cholecystitis and tumors may be prior therapeutic targets of this plant in the future.

Identification of potential key genes in esophageal adenocarcinoma using bioinformatics.

Esophageal adenocarcinoma (EAC) is the predominant pathological subtype of esophageal cancer in Europe and the USA. The present bioinformatics study analyzed a high-throughput sequencing dataset, GSE94869, to determine differentially expressed genes (DEGs) in order to identify key genes, biological processes and pathways associated with EAC. Functional enrichment analysis was performed using the Database for Annotation Visualization and Integrated Discovery. The co-expression network of the DEGs was established using Weighted Gene Co-Expression Network Analysis and visualized using Cytoscape. A Kaplan-Meier analysis based on The Cancer Genome Atlas (TCGA) database was used to identify prognosis-associated genes. Univariate and multivariate Cox proportional hazard models were used to identify genes with a prognostic value regarding relapse-free survival (RFS), while validation of the differential expression of prognosis-associated genes was performed using a box plot based on data from TCGA and another microarray dataset, GSE26886. A total of 130 DEGs, comprising 82 upregulated and 48 downregulated genes, were identified. The upregulated DEGs were significantly associated with extracellular matrix organization, disassembly, and the phosphoinositide-3 kinase/AKT, Rap1 and Ras signaling pathways, while the downregulated genes were associated with the Wnt signalling pathway. Subsequently, two co-expression modules were established and 20 hub genes were identified. The blue module was associated with the Rap1 signaling pathway, while the turquoise module was associated with the Ras and Rap1 signaling pathways. Among them, methyltransferase like 7B (METTL7B) was associated with RFS. Furthermore, the overexpression of METTL7B in EAC was successfully validated using data from TCGA and GSE26886. The present study identified key genes and provides potential biomarkers for the diagnosis and treatment of EAC.

Identification of prognostic risk factors for esophageal adenocarcinoma using bioinformatics analysis.

PURPOSE: Esophageal adenocarcinoma (EAC) is the most common type of esophageal cancer in Western countries. It is usually detected at an advanced stage and has a poor prognosis. The aim of this study was to identify key genes and miRNAs in EAC. METHODS: The mRNA microarray data sets GSE1420, GSE26886, and GSE92396 and miRNA data set GSE16456 were downloaded from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) and differentially expressed miRNAs (DEMs) were obtained using R software. Functional enrichment analysis was performed using the DAVID database. A protein-protein interaction (PPI) network and functional modules were established using the STRING database and visualized by Cytoscape. The targets of the DEMs were predicted using the miRecords database, and overlapping genes between DEGs and targets were identified. The prognosis-related overlapping genes were identified using Kaplan-Meier analysis and Cox proportional hazard analysis based on The Cancer Genome Atlas (TCGA) database. The differential expression of these prognosis-related genes was validated using the expression matrix in the TCGA database. RESULTS: Seven hundred and fifteen DEGs were obtained, consisting of 313 upregulated and 402 downregulated genes. The PPI network consisted of 281 nodes; 683 edges were constructed and 3 functional modules were established. Forty-four overlapping genes and 56 miRNA- mRNA pairs were identified. Five genes, FAM46A, RAB15, SLC20A1, IL1A, and ACSL1, were associated with overall survival or relapse-free survival. FAM46A and IL1A were found to be independent prognostic indicators for overall survival, and FAM46A, RAB15, and SLC20A1 were considered independent prognostic indicators for relapse-free survival. Among them, the overexpression of RAB15 and SLC20A1 and lower expression of ACSL1 were also identified in EAC tissues based on the expression matrix in the TCGA database. CONCLUSION: These prognosis-related genes and differentially expressed miRNA have provided potential biomarkers for EAC diagnosis and treatment.