Metachronous colorectal liver metastases (CRLM) had been the main cause of death in most colorectal cancer (CRC) patients after curative surgery. However, the clinical impact of the pre-existing metabolic dysfunction-associated steatotic liver disease (MASLD) on metachronous CRLM remains unclear, the present study aims to clarify this issue. This study retrospectively enrolled in 2155 CRC patients who underwent curative surgery at the Sixth Affiliated Hospital of Sun Yat-sen University from 2017 to 2018, including 300 cases with MASLD and 1855 cases of Control group. Cox regression and Kaplan–Meier curves were constructed to evaluate the influencing factors and cumulative incidence of recurrence (CIR) of metachronous CRLM. The pre-existing MASLD was an independent risk factor for metachronous CRLM in TNM stage 0–II CRC patients (HR, 2.696; 95

BACKGROUND:Colorectal cancer (CRC) is a leading cause of mortality in China, with metastasis significantly contributing to poor outcomes. Histopathological growth patterns (HGPs) in colorectal liver metastasis (CRLM) provide vital prognostic insights, yet the limited number of pathologists highlights the need for auxiliary diagnostic tools. Recent advancements in artificial intelligence (AI) have demonstrated potential in enhancing diagnostic precision, prompting the development of specialized AI models like COFFEE to improve the classification and management of HGPs in CRLM patients. METHODS:This study developed a Transformer-based deep learning model, COFFEE, for the precise classification of colorectal cancer subtypes using whole-slide images (WSIs) from 431 patients diagnosed with colorectal cancer liver metastasis. The model was pretrained using DINO on 1442 WSIs from the TCGA-COAD cohort, utilizing a Vision Transformer (ViT) architecture to extract 384-dimensional feature vectors from 256 × 256 pixel patches. The proposed model integrates a Transformer-based Multiple Instance Learning (TransMIL) framework, which effectively aggregates spatial and morphological information through multi-head self-attention and Pyramid Position Encoding Generator (PPEG) modules. This design enables efficient handling of large instance sequences within WSIs, allowing for accurate binary and four-class classification. The model was validated on 972 WSIs from a recent dataset, demonstrating its robustness and clinical applicability. After testing the model with internal and prospective cohorts, a direct comparison between conventional and AI-assisted pathology assessment was also performed. RESULTS:A total of 431 patients were included in three cohorts: training ( n = 297), testing ( n = 104), and prospective ( n = 30). Desmoplastic tumors were associated with longer overall survival (OS, 53.6 vs. 31.9 months, P = 0.002) and progression-free survival (PFS, 25.2 vs. 10.7 months, P < 0.001) compared to non-desmoplastic tumors. The COFFEE binary classification model achieved high predictive performance with area under the ROC curve (AUC) values of 0.961 in the training, 0.935 in the testing, and 1.000 in the prospective cohort. The four-class model also showed strong performance, with AUCs of 0.961 and 0.966 in the training and testing cohorts, and 0.985 in the prospective cohort. AI-assisted models helped junior pathologists achieve an accuracy of 94.7% (vs. 85.9%) and reduced diagnostic time by 36%, improving both accuracy and speed. CONCLUSION:This study developed an AI model for HGP classification in colorectal cancer liver metastasis, achieving high accuracy in both binary classification and four-class classification models. The model demonstrated potential for improving diagnostic precision and guiding post-surgery treatment strategies, with AI-assisted pathologists surpassing traditional methods in a prospective cohort.

Background: Bile leakage following hepatectomy is a challenging issue that poses difficulties for hepatobiliary surgeons. Endoscopic treatment is the primary method for managing this complication, although its effectiveness is still uncertain. Objectives: The aim of this study was to assess the effectiveness and risk factors of endoscopic treatment for bile leakage after hepatectomy. Design: A retrospective cohort study. Methods: A retrospective study was performed on 15 patients who experienced bile leakage following hepatectomy and were successfully treated with endoscopic retrograde cholangiography (ERCP). The primary focus was to examine how factors like the site of bile leakage, the position of biliary drainage, and late-set leakage impact early clinical success (ECS). Results: Out of the 15 cases, 10 successfully achieved ECS through endoscopic treatment. The highest success rate (80.0%, 4 out of 5) was seen when biliary drainage was performed by bridging, while drainage placed near the leakage site had a success rate of 75.0% (6 out of 8). The best outcomes occurred when bile leakage was located in the tertiary or higher branch bile ducts, with an 83.3% success rate (5 out of 6). Additionally, early-onset leakage had a higher ECS success rate compared to late-onset bile leakage (75.0% (6/8) vs 57.1% (4/7)). Among the 15 cases, 12 involved extensive liver resections of more than two segments, 14 included segments 7 or 8, 11 had received preoperative adjuvant therapy, and 9 had undergone transarterial chemoembolization (TACE). In cases with late-onset bile leakage, 6 out of 7 (85.7%) had undergone TACE. Bile leakage caused by TACE was more likely to be accompanied by bile duct-bronchial fistula, and the healing process took significantly longer. Conclusion: Endoscopic management of bile leakage following liver resection is both safe and effective, making it a preferred first-line treatment. Improved results can be obtained by placing a stent to bridge and drain near the leakage site. Extensive liver resection, resection involving segments 7 or 8 of the liver, and preoperative adjuvant therapies are significant risk factors for bile leakage, particularly TACE, which may lead to bile duct-bronchial fistulas and extended recovery periods.

Laparoscopic hepatectomy is a common treatment for colorectal cancer liver metastasis. Previously, a sufficient number of functional liver masses had to be maintained during laparoscopic hepatectomy, with a residual liver volume of >40% in cirrhotic patients and >30% in non-cirrhotic patients. The high incidence of complications such as bleeding, bile leakage, or liver failure due to the exposure and difficulty of the resection of specific liver segments such as S2 and S7 reduces the success rate of liver resection. At present, microwave ablation is mainly applied in the treatment of liver metastasis using a percutaneous approach, which makes it difficult to identify hidden parts or small lesions. For some liver segments, the percutaneous puncture of liver segment 7 (S7) is likely to pass through the thoracic cavity, and the percutaneous puncture of liver segment 2 (S2) adjacent to the diaphragm is likely to injure the diaphragm and heart; these issues restrict the application of percutaneous ablation in colorectal cancer liver metastasis. Considering multiple lesions, laparoscopic microwave ablation combined with hepatectomy was performed in this study. The location of the lesions was determined by contrast-enhanced ultrasound under laparoscopy, and small lesions that were difficult to detect before the operation were identified. For the scattered lesions, which had diameters less than 3 cm and were difficult to resect, ablation was adopted to substitute hepatectomy. This technique helped to more explicitly locate the tumors, simplified the operation procedures, reduced the risk of complications such as bleeding and bile leakage, shortened the operation time, accelerated the postoperative recovery, significantly improved the success rate of operation, and enhanced the clinical prognosis of colorectal cancer liver metastasis by surgical resection.

Cancer-associated fibroblasts (CAFs) are a kind of stromal cells in the cholangiocarcinoma (CCA) microenvironment, playing crucial roles in cancer development. However, the potential mechanisms of the interaction between CCA cells and CAFs remain obscure. This work investigated the role of circ_0020256 in CAFs activation. We proved circ_0020256 was up-regulated in CCA. High circ_0020256 expression facilitated TGF-β1 secretion from CCA cells, which activated CAFs via the phosphorylation of Smad2/3. Mechanistically, circ_0020256 recruited EIF4A3 protein to stabilize KLF4 mRNA and upregulate its expression, then KLF4 bound to TGF-β1 promoter and induced its transcription in CCA cells. KLF4 overexpression abrogated the inhibition of circ_0020256 silencing in TGF-β1/Smad2/3-induced CAFs activation. Furthermore, CCA cell growth, migration, and epithelial-mesenchymal transition were favored by CAFs-secreted IL-6 via autophagy inhibition. We also found circ_0020256 accelerated CCA tumor growth in vivo. In conclusion, circ_0020256 promoted fibroblast activation to facilitate CCA progression via EIF4A3/KLF4 pathway, providing a potential intervention for CCA progression.

Laparoscopic hepatectomy is a common treatment for colorectal cancer liver metastasis. Previously, a sufficient number of functional liver masses had to be maintained during laparoscopic hepatectomy, with a residual liver volume of >40% in cirrhotic patients and >30% in non-cirrhotic patients. The high incidence of complications such as bleeding, bile leakage, or liver failure due to the exposure and difficulty of the resection of specific liver segments such as S2 and S7 reduces the success rate of liver resection. At present, microwave ablation is mainly applied in the treatment of liver metastasis using a percutaneous approach, which makes it difficult to identify hidden parts or small lesions. For some liver segments, the percutaneous puncture of liver segment 7 (S7) is likely to pass through the thoracic cavity, and the percutaneous puncture of liver segment 2 (S2) adjacent to the diaphragm is likely to injure the diaphragm and heart; these issues restrict the application of percutaneous ablation in colorectal cancer liver metastasis. Considering multiple lesions, laparoscopic microwave ablation combined with hepatectomy was performed in this study. The location of the lesions was determined by contrast-enhanced ultrasound under laparoscopy, and small lesions that were difficult to detect before the operation were identified. For the scattered lesions, which had diameters less than 3 cm and were difficult to resect, ablation was adopted to substitute hepatectomy. This technique helped to more explicitly locate the tumors, simplified the operation procedures, reduced the risk of complications such as bleeding and bile leakage, shortened the operation time, accelerated the postoperative recovery, significantly improved the success rate of operation, and enhanced the clinical prognosis of colorectal cancer liver metastasis by surgical resection.

BACKGROUND:Serum protein induced by vitamin K absence or antagonist-II (PIVKA-II) is a promising biomarker for hepatocellular carcinoma (HCC) surveillance. AIM:To identify the contributing factors related to the abnormal elevation of PIVKA-II level and assess their potential influence on the performance of PIVKA-II in detecting HCC. METHODS:This study retrospectively enrolled in 784 chronic liver disease (CLD) patients and 267 HCC patients in Mengchao Hepatobiliary Hospital of Fujian Medical University from April 2016 to December 2019. Logistic regression and the area under the receiver operating characteristic curve (AUC) were used to evaluate the influencing factors and diagnostic performance of PIVKA-II for HCC, respectively. RESULTS:Elevated PIVKA-II levels were independently positively associated with alcohol-related liver disease, serum alkaline phosphatase (ALP), and total bilirubin (TBIL) for CLD patients and aspartate aminotransferase (AST) and tumor size for HCC patients (all P < 0.05). Serum PIVKA-II were significantly lower in patients with viral etiology, ALP ≤ 1 × upper limit of normal (ULN), TBIL ≤ 1 × ULN, and AST ≤ 1 × ULN than in those with nonviral disease and abnormal ALP, TBIL, or AST (all P < 0.05), but the differences disappeared in patients with early-stage HCC. For patients with TBIL ≤ 1 × ULN, the AUC of PIVKA-II was significantly higher compared to that in patients with TBIL > 1 × ULN (0.817 vs 0.669, P = 0.015), while the difference between ALP ≤ 1 × ULN and ALP > 1 × ULN was not statistically significant (0.783 vs 0.729, P = 0.398). These trends were then more prominently perceived in subgroups of patients with viral etiology and HBV alone. CONCLUSION:Serum PIVKA-II has better performance in detecting HCC at an early stage for CLD patients with normal serum TBIL.

Background: Tumor-associated macrophages (TAMs) play a dual role in tumors. However, the factors which drive the function of TAMs in cholangiocarcinoma remain largely undefined. Methods: SHH signaling pathway and endoplasmic reticulum stress (ERS) indicators were detected in clinical tissues and cholangiocarcinoma cell lines. TAMs were co-cultured with cholangiocarcinoma cells under conditions of hypoxia/normoxia. Polarized TAMs were counted by flow cytometry, and TGF-beta 1 levels in cell supernatants were detected by ELISA. The effects of glioma-associated oncogene GLI2 on TAMs themselves and cholangiocarcinoma cells were examined by conducting interference and overexpression assays. Results: The SHH signaling pathway and ERS were both activated in tumor tissues or tumor cell lines under conditions of hypoxia. In co-culture experiments, the presence of cholangiocarcinoma cells increased the proportion of M2-polarized TAMs and the secretion of TGF-beta 1 by TAMs, while knockdown of SHH expression reversed those increases. Overexpression of GLI2 in TAMS or stimulation of TAMS with Hh-Ag1.5 increased their levels of TGF-beta 1 expression. Furthermore, under co-culture conditions, interference with GLI2 expression in TAMs reduced the tumor cell migration, invasion, and ER homeostasis induced by Hh-Ag1.5-pretreated TAMs. Under conditions of hypoxia, the presence of cholangiocarcinoma cells promoted the expression of GLI2 and TGF-beta 1 in Tams, and in turn, TAMs inhibited the apoptosis and promoted the migration and invasion of cholangiocarcinoma cells. In vivo, an injection of cholangiocarcinoma cells plus TAMs contributed to the growth, EMT, and ER homeostasis of tumor tissue, while an injection of TAMs with GLI2 knockdown had the opposite effects. Conclusion: Cholangiocarcinoma cells regulated TAM polarization and TGF-beta 1 secretion via a paracrine SHH signaling pathway, and in turn, TAMs promoted the growth, EMT, and ER homeostasis of cholangiocarcinoma cells via TGF-beta 1.

Background:The accuracy of preoperative staging is crucial for cT4 stage gastric cancer patients. The aim of this study was to develop the radiomics model and evaluate its predictive potential for differentiating preoperative cT4 stage gastric cancer patients into pT4b and no-pT4b patients.Methods:A multicenter retrospective analysis of 704 gastric cancer patients with preoperative contrast-enhanced computed tomography (CE-CT) staging cT4 between January 2008 and December 2021. These patients were divided into the training cohort (478 patients, the Affiliated Hospital of Qingdao University) and validation cohort (226 patients, the Weihai Wendeng District People's Hospital). According to the pathological stage of the tumors, the patients were divided into pT4b or no-pT4b stage. In the training cohort, the clinical and radiomics features were analyzed to construct the clinical model, tri-phase radiomics signatures and nomogram. Two kinds of methods were employed to achieve dimensionality reduction: (I) the least absolute shrinkage and selection operator (LASSO); and (II) the minimum redundancy maximum relevance (mRMR) algorithms. We utilized Logistic regression, support vector machine (SVM), Decision tree and Adaptive boosted tree (AdaBoost) algorithms as the machine learning classifiers. The nomogram was constructed on the clinical characteristics and the Rad-score. The performance of the models was evaluated by receiver operating characteristic (ROC) area under the curve (AUC), Decision Curve Analysis (DCA) curve and calibration curve.Results:The 345 pT4b and 359 no-pT4b stage patients were included in this study. In the validation cohort, the AUC of the clinical model was 0.793 (95% CI: 0.732-0.855). The tri-phase radiomics features combined with the SVM algorithm was the best radiomics signature with an AUC of 0.862 (95% CI: 0.812-0.912). The nomogram was the best predictive model of all with an AUC of 0.893 (95% CI: 0.834-0.927). In the training and validation cohorts, the calibration curves and DCA curves of the nomogram showed satisfactory result.Conclusions:CE-CT-based radiomics nomogram offers good accuracy and stability in differentiating preoperative cT4 stage gastric cancer patients into pT4b and non-pT4b stages, which has a great clinical relevance for selecting the course of treatment for cT4 stage gastric cancer patients.

Objective To investigate the suppression of multidrug resistance-associated proteins 2 (MRP2) and the protein product induced by small interfering RNA (siRNA) in the multidrug-resistant (MDR) hepatocellular carcinoma (HCC) cell line HepG2/adriamycin (ADM) and the effect on MDR.Methods MDR HepG2/ADM cells were developed by exposing parental cells to stepwise increasing concentrations of ADM from 0.1 to 2.0 mg/L.MRP2 targeted small interfering RNA fragments were designed and synthesized,and transfected into MDR HepG2/ADM cells.The suppression of MRP2 and the protein product were detected by using real-time reverse transcription quantitative poiymerase chain reaction (RT-qPCR) and Western blotting at 24 h after transfection.Methyl thiazol tetrazolium (MTT) assay was used to determine drug sensitivity of HepG2/ADM cells before and after transfection based on the results of IC50.Results MTT assay showed that the IC50 values of HepG2/ADM against ADM,5-fluorouracil,vincristine,and oxaliplatin were 0.3204,3.8002,0.2014 and 0.1221 respectively.siRNA transfection significantly inhibited the expressiou of MRP2 mRNA and protein in HepG2/ADM cells ( P ＜ 0.05).After MRP2-siRNA transfection,the IC50 values of HepG2/ADM against ADM,5-fluorouracil,vincristine,and oxaliplatin were decreased significantly (0.1023,1.4417,0.0452 and 0.0268,respectively).Conclusion Silencing MRP2 genes by siRNA can reverse MDR of HepG2/ADM cells.MRP2 is closely related to MDR of HepG2/ADM HCC cells and silencing MRP2 might improve the sensitivity of resistant cells to chemotherapeutic drugs.

Objective Hepatocellular carcinoma (HCC) responds poorly to chemotherapy owing to multidrug resistance (MDR). This study was designed to observe the effect of small interfering RNA (siRNA) targeting MDR1 in modulating drug resistance and apoptosis. Methods HCC MDR cell lines,SMMC-7721/adriamycin ( ADM), were developed by exposing parental cells to stepwise increasing concentrations of ADM from 0. 1 to 2. 0 mg/L. After siRNA (25, 50, 75 nmol/L) transfection MDR1 mRNA expression levels were detected by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) at 24, 48, 72 h. methyl thiazol tetrazolium (MTT) assay was used to determine drug sensitivity 48 h after transfection. Flow cytometry (FCM) was employed to analyze cell apoptosis to ADM (0. 2mg/L). The expression of MDR1 protein was examined by Western blotting. Results MTT assay showed that the resistance index (RI) values of SMMC-7721/ADM cells against ADM, 5-Fu, VCR and oxaliplatin were 25.43, 68.32, 39. 17, 18. 31, and those after siRNA transfection were 5.01, 17.04, 4. 96, 1.62,respectively. QRT-PCR analysis demonstrated the MDR1 mRNA expression was decreased significantly in SMMC-7721/ADM cells after siRNA transfection (P＜ 0. 05 ). In addition, as compared with parental cells, MDR1 protein expression was apparently decreased in SMMC7721/ADM cells treated with siRNA (P ＜0. 05). Flow cytometry revealed significant apoptesis of the cells following siRNA transfection ( P ＜0. 05). Conclusion Inhibition of MDR1 by siRNA enhanced the selectively restored sensitivity to drugs.The MDR1 siRNA might represent a new therapeutic option for HCC.

Aberrant activation of the Hedgehog (Hh) signaling pathway has been reported in various cancer types including hepatocellular carcinoma (HCC). As a key effector of this signaling, Gli2 plays a crucial role in carcinogenesis, including the activation of genes encoding apoptosis inhibitors and cell‐cycle regulators. In this study, we examined the role of Gli2 proliferation and survival of HCC cells. First, the expression levels of Hh pathway components were detected in a subset of HCC cell lines. To establish the role of Gli2 in maintaining the tumorigenic properties of HCC cells, we developed small hairpin RNA (shRNA) targeting Gli2 and transfected it into SMMC‐7721 cell, which was selected with high level of Hh signaling expression. Next, effects of Gli2 gene silencing, on cell proliferation and on the expression of cell cycle‐related proteins were evaluated, then, whether down‐regulation of Gli2 renders HCC cell susceptible to TRAIL was examined in vitro. Knockdown of Gli2 inhibited cell proliferation and induced G1 phase arrest of cell cycle in SMMC‐7721 cell through down‐regulation of cyclin D1, cyclinE2, and up‐regulation of p21‐WAF1. Also, Gli2 gene siliencing sensitized SMMC‐7721 cell to tumor necrosis factor‐related apoptosis‐inducing ligand‐induced apoptosis by reducing the expression of the long and short isoform of c‐FLIP and Bcl‐2, and then augmented the activation of initiator caspases‐8/‐9 and effector caspases‐3, which induces PARP cleavage. In conclusion, our data suggest that Gli2 plays a predominant role in the proliferation and apoptosis resistance of HCC cells, and that knockdown of Gli2 may be a novel anticancer strategy for the treatment of HCC. J. Cell. Biochem. 112: 3140–3150, 2011. © 2011 Wiley Periodicals, Inc.

Objective To investigate the expression and significance of Hedgehog signaling pathway in human hepatocellular carcinoma cell strains.Methods Semi-quantitative RT-PCR was used for detections of Shh,.Ptch1,.Smo,.Gli1 and Gli2 mRNA,.and Western blot was used for detections of Gli2 protein in PLC/PRF/5、.HepG2 and SMMC-7721 hepatocellular carcinoma cell strains.Results Besides Shh,mRNA of Ptch1,.Smo,.Gli1,.Gli2 had expressions of different degree in PLC/PRF/5,HepG2 and SMMC-7721 cells.The mRNA expressions of Ptch1,Smo,Gli1,and Gli2 were significantly higher in PLC/PRF/5 and SMMC-7721 than that in normal adult liver cells,while mRNA expressions of Gli1 was no significant differences between HepG2 cells and adult liver cells.Gli2 protein was almost undetected in adult liver cells,.and was no significant differences between HepG2 cells and adult liver cells,.but Gli2 protein was highly expressed in PLC/PRF/5 and SMMC-7721 cell,particularly in SMMC-7721.Conclusion Abnormal activation of Hedgehog pathway may take part in development and progression of human HCC,.Gli2 may be an important biological marker of HCC and biological therapeutic target.

Background The prognosis for hepatocellular carcinoma (HCC) along with portal vein tumor thrombi (PVTT) is poor, and surgery has not been considered an option. Aims To compare the outcomes and the quality of life (QoL) of patients with HCC and PVTT who underwent hepatic resection and thrombectomy for tumor thrombi in the inferior vena cava and hepatic vein with total hepatic vascular exclusion to the patients who received only chemotherapy. Methods We retrospectively reviewed the medical records of patients who received hepatectomy and thrombectomy (n=65), and those who received only chemotherapy (n=50). The surgical outcomes, survival, and QoL that was determined using the Functional Assessment of Cancer Therapy-Hepatobiliary instrument were analyzed and compared. Results Patients who underwent surgery had a median overall survival of 17 months, compared with patients who underwent chemotherapy for 8 months (P<0.0001). Patients who underwent surgery had a median recurrence-free survival of 14 months, as compared with patients who underwent chemotherapy for 7 months (P<0.0001). The probabilities of 1-year recurrence in the surgery and chemotherapy groups were 27.7 and 70%, respectively (P<0.0001). The QoL total score of the surgery group was significantly higher than that of the control group (P<0.0001). Surgery was slightly, though significantly more cost-effective than chemotherapy based on the quality-adjusted life years. Conclusion Hepatectomy and thrombectomy using the total hepatic vascular exclusion, is a viable surgical management for patients with HCC and PVTT, and is associated with longer overall survival and recurrence-free survival and better QoL than chemotherapy alone.