OBJECTIVE: Gastric cancer (GC) is one of the most prevalent malignancies worldwide, and early detection is crucial for improving patient survival rates. We aimed to identify immune infiltrating cell-related biomarkers in early gastric cancer (EGC) progression.
METHODS: The GSE55696 and GSE130823 datasets with low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia (HGIN), and EGC samples were downloaded from the Gene Expression Omnibus database to perform an observational study. Immune infiltration analysis was performed by single sample gene set enrichment analysis and Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data. Weighted gene co-expression network analysis was used to explore the co-expression modules and genes, and further enrichment analysis was performed on these genes. A protein-protein interaction (PPI) network of these genes was constructed to identify biomarkers associated with EGC progression. Screened hub genes were validated by the rank sum test and reverse transcription quantitative polymerase chain reaction.
RESULTS: Immune scores were significantly elevated in EGC samples compared to LGIN and HGIN samples. The green-yellow module exhibited the strongest correlation with both immune score and disease progression. The 87 genes within this module were associated with the chemokine signaling pathways, the PI3K-Akt signaling pathways, leukocyte transendothelial migration, and Ras signaling pathways. Through PPI network analysis, the hub genes identified were protein tyrosine phosphatase receptor-type C (PTPRC), pleckstrin, CD53, CD48, lymphocyte cytosolic protein 1 (LCP1), hematopoietic cell-specific Lyn substrate 1, IKAROS Family Zinc Finger 1, Bruton tyrosine kinase, and Vav guanine nucleotide exchange factor 1. Notably, CD48, LCP1, and PTPRC showed high expression levels in EGC samples, with the remaining hub genes demonstrating a similar expression trend.
CONCLUSIONS: This study identified 9 immune cell-related biomarkers that may be actively involved in the progression of EGC and serve as potential targets for GC diagnosis and treatment.

UNASSIGNED: Early diagnosis of esophageal squamous cell carcinoma (ESCC) is critical for effective treatment and optimal prognosis; however, less study on serum biomarkers for the early ESCC detection has been reported. The aim of this study was to identify and evaluate several serum autoantibody biomarkers in early ESCC.
UNASSIGNED: We initially screened candidate tumor-associated autoantibodies (TAAbs) associated with ESCC by serological proteome analysis (SERPA) combined with nanoliter-liquid chromatography combined with quadrupole time of flight tandem mass spectrometry (nano-LC-Q-TOF-MS/MS), and the TAAbs were further subjected to analysis by Enzyme-linked immunosorbent assay (ELISA) in a clinical cohort (386 participants, including 161 patients with ESCC, 49 patients with high-grade intraepithelial neoplasia [HGIN] and 176 healthy controls [HC]). Receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic performance.
UNASSIGNED: The serum levels of CETN2 and POFUT1 autoantibodies which were identified by SERPA were statistically different between ESCC or HGIN patients and HC in ELISA analysis with the area under the curve (AUC) values of 0.709 (95%CI: 0.654-0.764) and 0.741 (95%CI: 0.689-0.793), 0.717 (95%CI: 0.634-0.800) and 0.703 (95%CI: 0.627-0.779) for detection of ESCC and HGIN, respectively. Combining these two markers, the AUCs were 0.781 (95%CI: 0.733-0.829), 0.754 (95%CI: 0.694-0.814) and 0.756 (95%CI: 0.686-0.827) when distinguishing ESCC, early ESCC and HGIN from HC, respectively. Meanwhile, the expression of CETN2 and POFUT1 was found to be correlated with ESCC progression.
UNASSIGNED: Our data suggest that CETN2 and POFUT1 autoantibodies have potential diagnostic value for ESCC and HGIN, which may provide novel insights for early ESCC and precancerous lesions detection.

Esophageal squamous cell carcinoma (ESCC) is thought to be started and developed by genes associated with inflammation. A cancer\'s ability to spread and grow can be aided by nuclear factor erythroid-2 related factor 2 (Nrf2) hyperactivation, which can also make a tumor more resistant to chemotherapy and radiation treatment. However, it is still unknown how Nrf2 gene expression affects ESCC prognosis and controls function throughout ESCC advancement.
The expression of Nrf2 and HO-1 in ESCC and precancerous esophageal precancerous lesions was analyzed, and their relationship with esophageal squamous cell carcinoma was analyzed.
Immunohistochemistry (IHC) was used to confirm the expression of Nrf2 and heme oxygenase-1 (HO-1) proteins in tissue microarrays from Chinese populations with ESCC. We looked at the connections between Nrf2/HO-1 expression and invading immune cells using the TIMER database.
Ethnicity and N stage are associated with Nrf2 overexpression. Differentiation, N stage, vascular invasion, distant metastasis, and American Joint Committee on Cancer (AJCC) staging are all associated with HO-1 overexpression. The expression of Nrf2 and HO-1 had a favorable correlation. Patients with elevated Nrf2 and HO-1 expression had lower progression-free survival (PFS) and overall survival (OS). In high-grade intraepithelial neoplasia, Nrf2 and HO-1 expression generally occurred, partially in low-grade intraepithelial neoplasia specimens, and rarely in normal mucosa. We further show that Nrf2 suppression is linked to higher immunological marker expression and lower immune cell infiltration.
The prognosis of ESCC may be improved by inhibiting the expression of Nrf2 and HO-1. A lack of immune cells was seen in ESCC with Nrf2 impairment.

OBJECTIVE: Gastric precancerous conditions such as atrophic gastritis (AG) and intestinal metaplasia (IM) are considered independent risk factors for gastric cancer (GC). The suitable endoscopic monitoring interval is unclear when we attempt to prevent GC development. This study investigated the appropriate monitoring interval for AG/IM patients.
METHODS: Totally, 957 AG/IM patients who satisfied the criteria for evaluation between 2010 and 2020 were included in the study. Univariate and multivariate analyses were used to determine the risk factors for progression to high-grade intraepithelial neoplasia (HGIN)/GC in AG/IM patients, and to determine an appropriate endoscopic monitoring scheme.
RESULTS: During follow-up, 28 AG/IM patients developed gastric neoplasia lesions including gastric low-grade intraepithelial neoplasia (LGIN) (0.7%), HGIN (0.9%), and GC (1.3%). Multivariate analysis identified H. pylori infection (P=0.022) and extensive AG/IM lesions (P=0.002) as risk factors for HGIN/GC progression (P=0.025).
CONCLUSIONS: In our study, HGIN/GC was present in 2.2% of AG/IM patients. In AG/IM patients with extensive lesions, a 1-2-year surveillance interval is recommended for early detection of HIGN/GC in AG/IM patients with extensive lesions.

Current evidence on the psychological impact of screening and diagnosis of esophageal cancer (EC) is limited and unclear.
This multicenter, population-based, prospective study was conducted in five high-incidence regions in China from 2017 to 2020. The screened participants were diagnosed as healthy, esophagitis, low-grade intraepithelial neoplasia (LGIN), high-grade intraepithelial neoplasia (HGIN), or EC based on pathological biopsy. The psychological impact of the screening was assessed by comparing anxiety and depression symptoms at baseline and follow-up.
A total of 1973 individuals were ultimately included, with an average follow-up of 22.2 months. The prevalence of anxiety and depression symptoms in screened population at baseline was 14.3% and 18.4%. The prevalence of anxiety and depression symptoms of screeners at follow-up declined (all p < 0.001). The anxiety (RR [95% CI]: 0.37 [0.30-0.46]) and depression (0.29 [0.24-0.36]) of screeners weakened over time, but the anxiety and depression symptoms was continuous for patients with HGIN and patients with EC. Compared with the participants classified as normal, the RRs(95% CI) of anxiety and depression symptoms were 2.20 (1.10-4.30) and 2.03 (1.07-3.86) for the patients with HGIN and 2.30 (0.82-6.20) and 3.79 (01.71-8.43) for the patients with EC.
The anxiety and depression symptoms of screeners weakened over time, except in patients with HGIN and EC, for whom it remained lasting and high. Psychological assistance and interventions are urgently needed for individuals who are ready for screening and for those diagnosed as having HGIN or EC.

High-grade intraepithelial neoplasia (HIN) is the precursor of oesophageal squamous cell carcinoma. The molecular and functional properties of HIN are determined by intrinsic origin cells and the extrinsic microenvironment. Yet, these factors are poorly understood.
We performed single-cell RNA sequencing of cells from HINs and adjacent tissues from the human oesophagus. We analysed the heterogeneity of basal layer cells and confirmed it using immunostaining. Aneuploid cells in HIN were studied using primary cell culture combined with karyotype analysis. We reconstructed the lineage relationship between tumour and normal populations based on transcriptome similarity. Integration analysis was applied to our epithelial data and published invasive cancer data, and results were confirmed by immunostaining and 3D organoid functional experiments. We also analysed the tumour microenvironment of HIN.
The basal layer contained two cell populations: KRT15high STMN1low and KRT15high STMN1high cells, which were located mainly in the interpapillary and papillary zones, respectively. The KRT15high STMN1low population more closely resembled stem cells and transcriptome similarity revealed that HIN probably originated from these slow-cycling KRT15high STMN1low cells. 3D Organoid experiments and RNA-sequencing showed that basal-cell features and the differentiation ability of the normal epithelium were largely retained in HIN, but may change dramatically in tumour invasion stage. Moreover, the tumour microenvironment of HIN was characterised by both inflammation and immunosuppression.
Our study provides a comprehensive single-cell transcriptome landscape of human oesophageal HIN. Our findings on the origin cells and unique microenvironment of HIN will allow for the development of strategies to block tumour progression and even prevent cancer initiation.

Little is known about esophageal high-grade intraepithelial neoplasia dominated by cytological atypia (HGINc). We aimed to elucidate the endoscopic features of HGINc compared with esophageal high-grade intraepithelial neoplasia dominated by architectural atypia (HGINa). All patients pathologically diagnosed as esophageal high-grade intraepithelial neoplasia after endoscopic submucosal dissection at our center between January 2018 and December 2019 were included in this study. According to the pathological diagnosis, the patients were divided into two groups: HGINa group and HGINc group. Basic characteristics and endoscopic information were collected in detail. Data were analyzed statistically. Binary logistic regression was performed and a predictive model for HGINc was established. Then we evaluated its predictive value and built a nomogram for clinical application. A total of 175 patients were included in this study (126 with HGINa and 49 with HGINc). Among 228 lesions found in all patients, there were 148 HGINa and 80 HGINc. The independent relevant factors for HGINc were tobacco and alcohol usage, color, and gross type. To predict risk of HGINc, a three-factor model (TFM) was established with a highest area under curve (AUC) as 0.869 (95% CI, 0.852, 0.939). When the cut-off value was set as 0.3569184, the diagnostic accuracy, sensitivity, specificity, positive predictive value, and negative predictive value for HGINc was 81.14%, 88.75%, 77.03%, 67.62%, and 92.68%, respectively. HGINc differs greatly in endoscopic features from HGINa in our study. It\'s important to reduce misdiagnosis that our model was established with good predictive value for clinical application.

Long non-coding (lnc) RNAs in circulating exosomes are a new class of promising cancer biomarkers; however, their expression in exosomes derived from gastric high-grade intraepithelial neoplasia (GHGIN) has not been reported. In the present study, differentially expressed (DE) lncRNAs were analyzed in the peripheral blood collected from 5 patients with GHGIN and 5 healthy donors using high-throughput sequencing. Reverse transcription-quantitative PCR analysis was performed on 6 randomly selected DE lncRNAs to validate the reliability of the sequencing results. The potential roles of the DE lncRNAs in GHGIN were investigated using Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genome (KEGG) pathway enrichment analyses. A total of 25,145 lncRNAs were identified in all the samples and 83 DE lncRNAs were further screened, including 76 upregulated and 7 downregulated DE lncRNAs. GO and KEGG analyses predicted that the DE lncRNAs played notable roles in \'protein/macromolecule glycosylation\', \'regulation of protein ubiquitination\', \'renin-angiotensin system\' and \'MAPK signaling pathways\'. A lncRNA-micro (mi)RNA-mRNA interaction network was constructed and used to perform association analyses. It was found that 83 lncRNAs were abnormally expressed in GHGIN, with some potential functions associated with gastric cancer. Furthermore, the lncRNA-miRNA-mRNA interaction network indicated that 7 DE lncRNAs may play a notable role in the occurrence and development of GHGIN. The results of the present study showed the expression profiles of lncRNAs in human GHGIN, elucidated some of the molecular changes associated with GHGIN and improved the understanding of the molecular mechanisms underlying GHGIN and gastric cancer.

OBJECTIVE: To analyse the current predictive value of isolated high-grade prostatic intraepithelial neoplasia (HGPIN) for clinically significant prostate cancer (csPCa) detection in repeat biopsies.
METHODS: A cohort of 293 men with isolated HGPIN detected in previous biopsies performed without multiparametric magnetic resonance imaging (mpMRI), and who underwent repeat biopsy within 1 to 3 years, was analysed. Pre-repeat biopsy mpMRI and guided biopsies to suspicious lesions (Prostate Imaging - Reporting and Data System [PI-RADS] ≥3) and/or and systematic biopsies were performed. Persistent prostate cancer (PCa) suspicion, defined as sustained serum prostate-specific antigen level >4 ng/mL and/or abnormal digital rectal examination, was present in 248 men (84.6%), and was absent in 45 men (15.4%). A control group of 190 men who had no previous HGPIN, atypical small acinar proliferation or HGPIN with atypia who were scheduled to undergo repeat biopsy due to persistent PCa suspicion were also analysed. csPCa was defined as tumours of Grade Group ≥2.
RESULTS: In the subset of 45 men with isolated HGPIN, in whom PCa suspicion disappeared, only one csPCa (2.2%) and one insignificant PCa (iPCa) were detected. csPCa was detected in 34.7% of men with persistent PCa suspicion and previous HGPIN, and in 28.4% of those without previous HGPIN (P =0.180). iPCa was detected in 12.1% and 6.3%, respectively (P =0.039). Logistic regression analysis showed that the risk of csPCa detection was not predicted by previous HGPIN: odds ratio (OR) 1.369 (95% confidence interval [CI] 0.894-2.095; P =0.149); however, previous HGPIN increased the risk of iPCa detection: OR 2.043 (95% CI 1.016-4.109; P =0.006).
CONCLUSIONS: The risk of csPCa in men with isolated HGPIN, in whom PCa suspicion disappears, is extremely low. Moreover, in those men in whom PCa suspicion persists, the risk of csPCa is not influenced by the previous finding of HGPIN. However, previous HGPIN increases the risk of iPCa detection. Therefore, repeat prostate biopsy should not be recommended solely because of a previous HGPIN.

Objective: To investigate the relationship between the expression of DNA methyltransferase 3b (DNMT3b) and the methylation of SEPT9 gene, and their application prospects in the diagnosis and treatment of colorectal cancer. Methods: Seventy-five cases of colorectal cancer and adjacent tissues, 68 cases of colorectal high-grade internal neoplasia tissues (referred to as precancerous tissues) and high-grade internal adjacent neoplasia tissues (referred to as adjacent precancerous tissues) were collected. Pyrosequencing was used to detect the methylationlevel of SETP9. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to examine the mRNA expressionof SEPT9 and immunohistochemistry(IHC) was applied to detect the protein expressions of SETP9 and DNMT3b. Liposome-mediated method was used to transfect DNMT3b siRNA and negative control siRNA into HT-29 cells. Five groups including DNMT3b siRNA 15 nmol/L group, DNMT3b siRNA 30 nmol/L group, negative control siRNA 15 nmol/L group, negative control siRNA 30 nmol/L group and blank control group were set up. Pyrosequencing was applied to determine the methylation level of SEPT9 and mRNA expression of DNMT3b in each group. Results: The methylation rates of SEPT9 gene in colorectal cancer tissues, adjacent tissues, precancerous tissues and adjacent precancerous tissues were (76.8±6.5)%, (14.4±2.6)%, (34.6±5.0)% and (7.4±1.2)%, respectively, which was highest in colorectal cancer tissue (P<0.001). The relative expressions of SEPT9 mRNA were 0.18±0.03, 0.89±0.41, 0.69±0.41 and 1.01±0.21, respectively, which was lowest in colorectal cancer tissue (P<0.001), while there were no statistically significant differences in adjacent tissues, precancerous tissues and adjacent precancerous tissues (P>0.05). The positive rates of SEPT9 protein expression were 12.0% (9/75), 53.3% (40/75), 55.1% (38/69) and 62.3% (43/69), which was lowest in the colorectal cancer tissue (P<0.001), while there were no statistically significant differences in the adjacent group, precancerous group and adjacent precancerous group (P>0.016 7). The positive rates of DNMT3b protein expression were 56.3% (45/75), 26.7% (20/75), 46.4% (32/69) and 33.3% (23/69), respectively, which was highest in colorectal cancer tissue (P<0.001), while without statistically significant difference from the precancerous tissue (P>0.016 7). Experiments in vitro showed that DNMT3b mRNA expression was lowest in DNMT3b siRNA 30 nmol/L group among five groups and was statistically different from other groups (all P<0.05). Meanwhile, the methylationrate of SEPT9 gene was lowest in this group, but without statistically significant difference from the DNMT3b siRNA 15 nmol/L group (P>0.05). Conclusions: The expression of DNMT3b is significantly correlated with the methylation level of SEPT9 gene in different stages of colorectal cancer. The high expression of DNMT3b may be an important molecular event before SEPT9 gene methylation and it may have an important potential application value in the diagnosis and treatment of early colorectal cancer.
目的： 探讨DNA甲基转移酶3b(DNMT3b)表达与SEPT9基因甲基化的关系，以及二者在结直肠癌诊治中的应用前景。 方法： 收集75例结直肠癌及癌旁组织、68例结直肠高级别内瘤变组织(简称癌前病变组织)和高级别内瘤变旁组织(简称癌前病变旁组织)，采用焦磷酸测序检测SETP9基因甲基化水平，采用实时荧光定量聚合酶链反应检测SEPT9 mRNA的表达，采用免疫组化方法检测SETP9和DNMT3b的蛋白表达。采用脂质体介导法，将DNMT3b siRNA和阴性对照siRNA转染HT-29细胞，设DNMT3b siRNA 15 nmol/L组、DNMT3b siRNA 30 nmol/L组、阴性对照siRNA 15 nmol/L组、阴性对照siRNA 30 nmol/L组和空白对照组。采用焦磷酸测序检测各组SEPT9基因甲基化和DNMT3b mRNA表达水平。 结果： 结直肠癌组织、癌旁组织、癌前病变组织和癌前病变旁组织中，SEPT9基因甲基化率分别为(76.8±6.5)%、(14.4±2.6)%、(34.6±5.0)%和(7.4±1.2)%，结直肠癌组织最高(P<0.001)；SEPT9mRNA的相对表达量分别为0.18±0.03、0.89±0.41、0.69±0.41和1.01±0.21，结直肠癌组织最低(P<0.001)，而癌旁组织、癌前病变组织及癌前病变旁组织差异无统计学意义(P>0.05)；SEPT9蛋白表达阳性率分别为12.0%(9/75)、53.3%(40/75)、55.1%(38/69)和62.3%(43/69)，结直肠癌组最低(P<0.001)，而癌旁组、癌前病变组及癌前病变旁组差异无统计学意义(P>0.016 7)；DNMT3b蛋白表达阳性率分别为56.3%(45/75)、26.7%(20/75)、46.4%(32/69)和33.3%(23/69)，结直肠癌组最高(P<0.001)，但与癌前病变组差异无统计学意义(P>0.016 7)。体外实验显示，空白对照组、阴性对照siRNA 15 nmol/L组、阴性对照siRNA 30 nmol/L组、DNMT3b siRNA 15 nmol/L组和DNMT3b siRNA 30 nmol/L组中，DNMT3b siRNA 30 nmol/L组DNMT3b mRNA表达量最低，与其他各组差异均有统计学意义(均P<0.05)，SEPT9基因甲基化率最低，但与DNMT3b siRNA 15 nmol/L组差异无统计学意义(P>0.05)。 结论： 在结直肠癌病变进程的不同阶段，DNMT3b的表达与SEPT9基因甲基化水平有明显的相关性，DNMT3b高表达可能是SEPT9基因甲基化前重要的分子事件，在结直肠癌早期病变诊治中具有重要的潜在应用价值。.