Esophageal squamous cell carcinoma(ESCC)often exhibits a high incidence of multifocality.The dis-tinction between multiple primary and metastatic tumor origin is a prerequisite for precise diagnosis and treatment.Based on histopathological diagnosis,molecular pathological diagnostic indicators and technologies utilizing multi-omics approaches(DNA,RNA,protein)have been gradually identified.This article systematically reviews recent advance-ments in pathological diagnostic techniques for distinguishing the origin of multifocal ESCC and provides future perspec-tives,aiming to offer references for precise diagnosis.

Purpose This study aimed to investigate the expression and clinical significance of TLDC2 in colorec-tal adenocarcinoma.Methods Data from the human protein atlas(HPA)and the cancer genome atlas(TCGA)indi-cated that TLDC2 was highly expressed in colorectal adenocarcinoma.We further analyzed the expression of TLDC2 in 400 colorectal adenocarcinomas and 447 other solid tumors using tissue microarrays and immunohistochemical(IHC)staining.Result The positive expression rate of TLDC2 was significantly higher than that of SATB2 in colorectal ade-nocarcinomas(96.5％vs 87.0％,P＜0.000 1).TLDC2 positivity exceeded that of SATB2 in both low-or high-grade colorectal adenocarcinoma(99.4％vs 88.7％,P＜0.000 1;83.3％vs 79.2％,P=0.669 9).In addition,the expression of TLDC2 and SATB2 was evaluated in 447 cases of other types of solid tumors.TLDC2 was expressed in neuroendocrine tumors as well as in gastric and appendiceal adenocarcinomas,whereas SATB2 was detected in a small number of melanomas,ovarian cancers,breast cancers and gallbladder cancers.The positive and specificity of TLDC2 for colorectal adenocarcinoma were 97％(95％CI=0.94-0.98)and 85％(95％CI=0.81-0.88),respectively.Combined detection of TLDC2 and SATB2 yielded a sensitivity of 96％(95％CI=0.93-0.97)and a specificity of 93％(95％CI=0.90-0.95).Conclusion Analysis of large-scale datasets and IHC staining demonstrated that TLDC2 is a highly sensitive and specific biomarker for colorectal adenocarcinoma.

Immunohistochemistry(IHC)controls are essential tools used in IHC experiments to monitor the exper-imental process and ensure the accuracy and reliability of results.These controls typically contain known specific anti-gen or antibody concentrations and simulate the matrix of actual samples to evaluate the performance of the experimental method.To address key issues in the clinical application of IHC detection technology,Pathological Technical Expert Group of Medical Technician Committee of Chinese Medical Doctor Association,Standardization Department of China Association of Pathology Equipment,Pathological Technology Group of Pathological Committee of China Anticancer As-sociation,and Pathological Technology Group of Pathology Committee of Chinese Research Hospital Association organ-ized experts from related fields to conduct indepth discussions and reached this consensus on the concept,design,placement,preparation,and interpretation of IHC controls.The purpose is to provide guidance for clinical laboratories in conducting IHC testing and quality control,while simultaneously enhancing the accuracy and reliability of test re-sults.

Purpose To investigate the clinicopathologic,molecular genetic features of metastatic adenoid cystic carcinoma(ACC)of the liver.Methods The histological morphological characteristics,immunophenotype and prog-nosis of metastatic ACC in 4 cases of liver biopsy specimens collected were retrospectively analyzed,and the genetic characteristics were analyzed by fluorescence in situ hybridization(FISH).Results There were 3 male patients and 1 female patients,ranging in age from 41 to 66 years,with an average age of 52.5 years.Pathologically,the tumor cells showed a variety of tubular,cribriform or solid structures.The tumor cells had a bland morphology,composed of two layers of cells:the luminal cells with eosinophilic cytoplasm and round nuclei,and the peripheral cell with deeply stained,triangular or irregular nuclei.Immunohistochemistry showed that CK7,CK19 and CD117 were positive in lu-minal cells.p63,Calponin,SMA and CK5/6 were positive in peripheral cells.TTF-1,Napsin A,CD10,CD56 and CRP were negative,and Ki67 proliferation index was 8％-20％.4 patients had a history of adenoid cystic carcinoma of lung or submandibular gland.FISH results showed that MYB gene rearrangement in case 3 and MYB gene amplifica-tion in case 2.All 4 patients were diagnosed with liver metastatic ACC.After 27 to 65 months of follow-up,all the 4 patients had multiple systemic metastases such as bone and lymph nodes,of which 1 patient died of disease 36 months after surgery,and 3 patients survived with tumor.Conclusion The histological morphology of metastatic ACC in the liver is similar to that of benign and malignant tumors and metastatic tumors of the primary bile duct in the liver,espe-cially when it presents as small tubular form.The characteristic morphology of the two layers of epithelia should be carefully observed during diagnosis,and the patient history should be asked.Combined with the histochemical results and molecular detection,it can distinguish from primary and metastatic tumors such as hepatic bile duct adenoma,in-trahepatic small bile duct type cholangiocarcinoma,hepatic metastatic epithelial-myoepithelial carcinoma and metastatic adenocarcinoma.

Purpose To investigate how IL-6-mediated monocytes/macrophages suppress the effector function of natural killer(NK)cells through the PD-L1/PD-1 axis in acute-on-chronic liver failure(ACLF).Methods HE stai-ning and Masson's trichrome staining were used to assess hepatic inflammatory infiltration and fibrosis.Immunofluores-cence staining was performed to detect IL-6 expression in hepatic macrophages.Flow cytometry was applied to evaluate the phenotypes and functions of monocytes/macrophages and NK cells.Recombinant IL-6 was used to treat peripheral blood mononuclear cells,and PD-L1 expression on monocytes was examined.Monocytes and NK cells were co-cul-tured,and after PD-1 blockade,NK cell effector function was evaluated.Results The ACLF patients,the degree of hepatic fibrosis increased with higher serum IL-6 level.Hepatic macrophages secreted more IL-6,and the circulating monocytes showed an increased percentage[(9.89±0.65)％],absolute count[(0.58±0.04)× 106/mL],prolif-erative capacity[(36.65±6.810)％],and IL-6 secretion[(16.97±2.387)％].In ACLF mice,hepatic inflamma-tory infiltration and fibrosis were aggravated,and IL-6+macrophages exhibited enhanced pro-inflammatory activity.IL-6 had mediated the upregulation of PD-L1 expression in monocytes,while NK cells showed increased PD-1 expression and impaired effector function.When monocytes and NK cells were co-cultured,PD-1 blockade restored,NK cell ef-fector functions.Conclusion In ACLF,elevated IL-6 levels were associated with hepatic fibrosis and disease severi-ty.IL-6 mediated monocytes-induced suppression of NK cell effector functions via the PD-L1/PD-1 axis,suggesting that IL-6 plays a critical role in ACLF-related immune dysregulation.

Cancer-associated fibroblasts(CAFs),as major components of the tumor microenvironment,play a pivotal role in therapeutic resistance in gastric cancer.Evidence shows that CAFs contribute to resistance through vari-ous multiple mechanisms,including exosomal signaling,metabolic reprogramming,extracellular matrix remodeling,and immune modulation.Although the regulatory network of CAFs is highly complex and the clinical translation of relat-ed therapeutic strategies remains challenging,precise targeting CAFs offers a promising avenue for overcoming drug re-sistance and improving patient outcomes.This review summarizes the molecular mechanisms by which CAFs mediate resistance and signaling pathways,biomarker-based immunotherapy,and emerging combination regimens,aiming to provide novel insights for gastric cancer treatment.

Purpose This study aimed to investigate the expression,function,and molecular mechanisms of ZNF384 in esophageal squamous cell carcinoma(ESCC),as well as its role in tumor progression and chemoresistance.Methods The expression of ZNF384 in ESCC cell lines and tissues was assessed using RT-qPCR.Correlations with TNM stage,invasion depth,lymph node metastasis,and prognosis were evaluated.In vitro assays were performed to examine the effects of ZNF384 on ESCC cell proliferation,migration,invasion,and chemosensitivity.Dual-luciferase reporter assays were conducted to determine the interaction between ZNF384 and FZD3,and to assess the activation of the Wnt signaling pathway.Results ZNF384 expression was significantly upregulated in ESCC cell lines and tissues(P＜0.01).Elevated ZNF384 expression was associated with advanced TNM stage,greater invasion depth,lymph node metastasis,and poor prognosis(P＜0.05).Functional assays demonstrated that ZNF384 overexpression promo-ted ESCC cell proliferation,migration,and invasion(all P＜0.01),whereas ZNF384 knockdown inhibited these processes and enhanced chemosensitivity to cisplatin(all P＜0.01).Mechanistic studies showed that ZNF384 directly bound to the FZD3 promoter,upregulated FZD3 expression,and activated the Wnt signaling pathway(P＜0.05).Overexpression of FZD3 partially reversed the inhibitory effects of ZNF384 knockdown on cell malignancy and chemore-sistance(P＜0.05).Conclusion ZNF384 promotes ESCC progression and reduces chemosensitivity through activa-tion of the FZD3/Wnt signaling pathway,suggesting its potential as a therapeutic target in ESCC.

Purpose To investigate the expression of LAMB3 and ITGB4 in esophageal squamous cell carcinoma(ESCC)and analyze their associations with clinicopathological features.Methods Bioinformatics was used to assess the expression of LAMB3 and ITGB4 in pan-cancer and ESCC.Immunohistochemistry was performed to evaluate their expression and distribution in ESCC tissues,and correlations clinicopathological features were analyzed.Follow-up data were collected to construct Kaplan-Meier survival curves,and Cox regression analysis was proformed to determine their prognostic significance.Results Bioinformatics analysis showed that LAMB3 and ITGB4 were highly expressed in ES-CC tissues(P＜0.001).Immunohistochemistry confirmed that both proteins were localized in the cytoplasm and membrane of tumor cells.High LAMB3 expression was significantly associated with poor differentiation(P＜0.001),lymph node metastasis(P=0.015),and T staging(P＜0.001).High ITGB4 expression was significantly correlated with poor differentiation(P=0.004)and advanced T staging(P=0.004).Cox regression analysis identified high ex-pression of LAMB3 and ITGB4 as independent prognostic factors for overall survival[HR=4.97(95％CI:2.73-9.02);HR=2.33(95％CI:1.36-3.99)].Conclusion LAMB3 and ITGB4 are highly expressed in ESCC.High LAMB3 expression is associated with tumor differentiation,lymph node metastasis,and T staging,while high ITGB4 expression is correlated with tumor differentiation and T staging.Patients with high expression of LAMB3 or ITGB4 have poorer overall survival,suggesting that these proteins may serve as prognostic biomarkers for unfavorable outcome in ESCC.

Purpose The current study aims to elucidate the interrelationships among IREB2,FBXL5,iron ho-meostasis,and the therapeutic efficacy of neoadjuvant chemotherapy.Methods A total of 97 samples,classified into colorectal cancer neoadjuvant chemotherapy-resistant and-sensitive groups,along with their corresponding paracancer-ous normal mucosa were collected.The expression levels of FBXL5,IREB2,TFRC and FTH1 were detected by immu-nohistochemistry,Werstern blot and RT-qPCR.The contents of ferroptosis-related markers Fe2+,MDA,ROS and GSH were detected by applying the kit,and the levels of these markers were analyzed.The relationship between each factor in different colorectal cancer tissues and tumor regression rate and prognosis of neoadjuvant chemotherapy were ana-lyzed.Results(1)The expression of IREB2,FBXL5,TFRC and FTH1 in colorectal cancer was higher than that in normal intestinal mucosa(P＜0.05),and the expression of FBXL5 and FTH1 in colorectal cancer drug-resistant group was lower than that in the sensitive group,whereas the expression of IREB2 and TFRC was higher than that in the sen-sitive group(P＜0.05);(2)The correlation analysis showed a positive correlation between the expression of IREB2 and TFRC in the drug-resistant group,and a negative correlation with the expression of FBXL5 and FTH1 in the drug-resistant group.TFRC expression in the colorectal cancer resistance group were positively correlated(P＜0.05)and negatively correlated with the expression of FBXL5 and FTH1(P＜0.05);(3)the content of Fe2+and GSH was high-er than that of the sensitivity group in the colorectal cancer resistance group,and the level of ROS was lower than that of the sensitivity group(P＜0.05);(4)Fe2+was positively correlated with the expression of IREB2,TFRC,and neg-atively correlated with the expression of FBXL5 and FTH1,and was negatively correlated with the expression of FBXL5 and FTH1.FTH1 expression were both negatively correlated(P＜0.05);(5)tumor regression rate was positively cor-related with the expression of FBXL5 and FTH1,and negatively correlated with the expression of IREB2 and TFRC,as well as positively correlated with the level of ROS,and negatively correlated with Fe2+and GSH(P＜0.05);(6)The expression of IREB2 and TFRC was positively correlated with tumor diameter and lymph node metastasis,and the ex-pression of FBXL5 and FTH1 was negatively correlated with tumor diameter and lymph node metastasis,and the expres-sion of FBXL5 was also negatively correlated with the depth of tumor infiltration(P＜0.05);(7)Kaplan-Meier analy-sis showed that lymph node metastasis,FBXL5,IREB2,TFRC,FTH1,and TRG grading were closely related to the prognosis of colorectal cancer patients(P＜0.05).Cox multivariate regression analysis indicated that lymph node me-tastasis,high expression of IREB2 and TFRC,low expression of FBXL5 and FTH1,and low tumor regression grade(TRG)were risk factors for the ineffectiveness of neoadjuvant chemotherapy in colorectal cancer patients.Conclusion FBXL5 and IREB2 are not only associated with high iron homeostasis,but also closely related to the efficacy and prognosis of neoadjuvant chemotherapy for colorectal cancer.In the future,they may become new targets for the treat-ment of colorectal cancer and improve the prognosis of patients.

Purpose To investigate the clinicopathological characteristics of the gastric oxyntic gland adenoma(GOGA).Methods We collected 18 samples of GOGA,histopathological features and immunohistochemical staining were assessed.Main features of pathological diagnosis,treatment methods and follow-up were retrospectively analyzed.Results There were 18 patients,including 9 females and 9 males,aged from 36 to 86 years old.The endoscopic im-age showed a flat lesion with whitish in color or a polypoid protrusions.The size ranged from 0.3 cm to 0.8 cm.Hema-toxylin and eosin staining showed irregular glandular structures in the mucosal lamina propria,with branched and anas-tomosed patterns.The tumour demonstrating composed of chief cells hyperplasia with mild nuclear atypia.All lesions were confined to the mucous lamina propria.There was no atrophic within the peripheral gastic mucosa.Immunohisto-chemical examination showed positive for Pepsinogen-Ⅰ and MUC6.Gene mutation were analyzed in 2 cases using next generation sequence technology,and no KRAS and GNAS mutation had been detected.Endoscopic surgical treatment was performed in 11 cases,and biopsy forceps removal was carried out in 7 cases.No recurrence or metastasis was ob-served during the follow-up period of 1 to 58 months.Conclusion GOGA is a rare lesion,and appears to behave bio-logically benign.A full understanding of its histological morphology and biological behavior can improve the diagnostic ability of clinincans,and facilitate further research in the future.