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Background: Abnormal glycosylation is associated with tumors. The clinical value of serum glycans in assessing progression of hepatocellular carcinoma (HCC) patients remains a challenge., Methods: A study dynamically comparing levels of fifteen lectin-specific glycans between preoperative and postoperative serum of 65 HCC patients was conducted via lectin biochip technology. Multivariable logistic regression analysis was used to address associations between serum glycan levels and clinicopathological characteristics. Kaplan-Meier analysis was used to evaluate the impacts of serum glycan levels on overall survival (OS) and progression-free survival (PFS) of the HCC patients., Results: HCC patients presented significantly higher levels of the lectin-specific glycans in preoperative serum than disease-free individuals (p < 0.001 - p = 0.029), except ConA. The glycans in preoperative sera were significantly related to tumor size, pTNM, metastasis, BCLC stage, portal hypertension (PHT), and platelet count (PLT), respectively (p < 0.05). Multivariate logistic analyses indicated that tumor size and pTNM independently impact on glycan-specific lectins either LTL, UEA-I, VVL, NPL, WGA, PNA, MAL-I, SNA, or PHA-L (p = 0.003 - p = 0.044); BCLC stage and PLT were independent factors influencing the serum glycans recognizable DSA (p = 0.024) and SNA (p = 0.050), respectively. Surgical excision of tumor mass significantly reduced glycan levels in sera. Tumor differentiation, albumin, and ABO type significantly revealed independent influence on glycan-specific lectins, such as RCA-I (p = 0.024), VVL (p = 0.024), and Con A (p = 0.026) in the postoperative serum. HCC patients with high levels of VVL-binding glycans significantly benefited from a longer OS time (p = 0.016, HR: 0.460, 95% CI: 0.237-0.892) and a better PFS time (p = 0.004; HR: 0.435, 95% CI: 0.237-0.799), respectively., Conclusion: Serum glycans could reflect surgical outcomes in at-risk patients and become valuable biomarkers in evaluating the progression of HCC patients., (© 2024 The Author(s). Cancer Medicine published by John Wiley & Sons Ltd.)

Background: CTCs play a critical role in the diagnosis and prognosis of liver cancer. However, there are few studies on whether different types of CTCs can predict the prognosis in patients with HCC following LT., Methods: Retrospective data including CTCs detected by the CanPatrolTM platform combined with RNA-ISH were collected and analyzed on 56 patients from December 2016 to December 2019 at the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong Province, China., Results: During the study period, fifty-six patients (51 males, 5 females) were included with an mean age of 52 ± 9 years. The 1-, 2- and 3-year recurrence rates of postoperative interstitial CTC-positive and CTC-negative groups were 21.7% vs 10.8%, 37.5% vs 10.8% and 55.5% vs 10.8%, confirming a statistically significant difference between the 2 groups (p = 0.044). The 1-, 2- and 3-year recurrence rates of the increasing interstitial CTCs group were 25.2%, 36.9% and 66.9%, while 12.6%, 24.4% and 24.4% in the decreasing and unchanged group, indicating a significant difference (p = 0.038)., Conclusion: CanPatrolTM platform presents a superior analytical sensitivity, and may be used as a dynamic monitoring tool for CTCs. And interstitial CTCs which are more aggressive and metastatic caused by EMT can be regarded as a predictor of post-transplant tumor recurrence after LT for HCC., (Copyright © 2021. Published by Elsevier Taiwan LLC.)

Immune checkpoint inhibitors are increasingly used in the treatment of various solid tumors, including hepatocellular carcinoma (HCC). For liver transplant recipients, the safety of using immune checkpoint inhibitors before or after transplantation remains to be further explored. Former reports were mainly about posttransplant use of immune checkpoint inhibitors resulting in allograft rejection. Here we present one HCC patient who received toripalimab (an immune checkpoint inhibitor currently in phase 3 clinical trial for HCC) therapy before undergoing liver transplantation. He finally suffered fatal acute hepatic necrosis which is likely to be related to the acute immune rejection caused by the pretransplant use of toripalimab., (Copyright © 2021. Published by Elsevier B.V.)

Purpose: Microvascular invasion (MVI) is a valuable predictor of survival in hepatocellular carcinoma (HCC) patients. This study developed predictive models using eXtreme Gradient Boosting (XGBoost) and deep learning based on CT images to predict MVI preoperatively., Methods: In total, 405 patients were included. A total of 7302 radiomic features and 17 radiological features were extracted by a radiomics feature extraction package and radiologists, respectively. We developed a XGBoost model based on radiomics features, radiological features and clinical variables and a three-dimensional convolutional neural network (3D-CNN) to predict MVI status. Next, we compared the efficacy of the two models., Results: Of the 405 patients, 220 (54.3%) were MVI positive, and 185 (45.7%) were MVI negative. The areas under the receiver operating characteristic curves (AUROCs) of the Radiomics-Radiological-Clinical (RRC) Model and 3D-CNN Model in the training set were 0.952 (95% confidence interval (CI) 0.923-0.973) and 0.980 (95% CI 0.959-0.993), respectively (p = 0.14). The AUROCs of the RRC Model and 3D-CNN Model in the validation set were 0.887 (95% CI 0.797-0.947) and 0.906 (95% CI 0.821-0.960), respectively (p = 0.83). Based on the MVI status predicted by the RRC and 3D-CNN Models, the mean recurrence-free survival (RFS) was significantly better in the predicted MVI-negative group than that in the predicted MVI-positive group (RRC Model: 69.95 vs. 24.80 months, p < 0.001; 3D-CNN Model: 64.06 vs. 31.05 months, p = 0.027)., Conclusion: The RRC Model and 3D-CNN models showed considerable efficacy in identifying MVI preoperatively. These machine learning models may facilitate decision-making in HCC treatment but requires further validation.

Background: Many studies confirm that anatomical resection was associated with favorable oncologic outcomes for patients with HCC who had preserved as much of the remnant liver tissue as possible. 1,2 In recent years, laparoscopic liver resection has been widely extended from minor resection to complex hepatectomy, 3 However, surgery on tumors located in the posterosuperior segment remains a demanding procedure regardless of the extent of resection. 4 Laparoscopic anatomical segment VII resection has one of the highest difficulty scores based on the tumor location due to poor accessibility, hard to exposure, and difficulty in obtaining sufficient surgical margins. 5,6 Here, we report a totally laparoscopic anatomical VII resection using the Glissonian approach with indocyanine green dye fluorescence., Methods: A 74-year-old man with a body mass index of 31.9 kg/m 2 suffered from HBV-related cirrhosis was admitted to our institution. The preoperative Gd-EOB-DTPA MRI showed a 2.7-cm HCC located in segment VIII. The preoperative AFP is 3431 ng/ml. A true anatomical segmentectomy was performed by using selective occlusion of segment VII Glissonian pedicle, which was identified from the liver hilum. Indocyanine green (ICG) dye demarcation was used as a guidance during parenchymal transection., Results: The operative time was 270 min with an estimated blood loss of 200 mL. The postoperative course was uneventful. Drainage tube was pulled out on the fourth day. The pathology confirmed the diagnosis of hepatocellular carcinoma and the surgical margin was negative. The patient was discharged on the 8th day after operation., Conclusions: Totally laparoscopic anatomical segment VII resection is a technically challenging operation. Advanced laparoscopic skills are necessary to complete such a difficult procedure safely. Glissonian approach and ICG fluorescence imaging guide parenchyma resection could be help.

Background/aims: Patient selection is critically important in improving the outcomes of liver transplantation for hepatocellular carcinoma. The aim of the current study was to identify biochemical measures that could affect patient prognosis after liver transplantation., Methods: A total of 119 patients receiving liver transplantation for hepatocellular carcinoma were used to construct a model for predicting recurrence. The results were validated using an independent sample of 109 patients from independent hospitals. All subjects in both cohorts met the Hangzhou criteria., Results: Analysis of the discovery cohort revealed an association of recurrence with preoperative fibrinogen and AFP levels. A mathematical model was developed for predicting probability of recurrence within 5 years: Y = logit(P) = -4.595 + 0.824 ×fibrinogen concentration (g/L) + 0.641 × AFP score (1 for AFP<=20ng/ml, 2 for 20 400ng/ml). At a cutoff score of -0.85, the area under the curve (AUC) was 0.819 in predicting recurrence (vs. 0.655 when using the Milan criteria). In the validation cohort, this model had reasonable performance in predicting 5-year overall survival (68.8% vs. 28.1% in using the -0.85 cutoff, p< 0.001) and disease-free survival (65.7% vs. 25.9%, p< 0.001). The sensitivity and specificity were 77.0% and 62.5%, respectively. The AUC of this newly developed model was similar to that with the Milan criteria (0.698 vs. 0.678). Surprisingly, the DFS in patients with score <= -0.85 under this model but not meeting the Milan criteria was similar to that in patients meeting the Milan criteria (53.8% vs. 60.0%, p=0.380)., Conclusions: Preoperative AFP and fibrinogen are useful in predicting recurrence of hepatocellular carcinoma after liver transplantation., (© 2018 The Author(s). Published by S. Karger AG, Basel.)

Background: Whether hepatic resection (HR) could be performed for patients with Barcelona Clinic Liver Cancer (BCLC) B/C stage hepatocellular carcinoma (HCC) is controversial, and the safety and clinical value of HR combined with antiviral therapy for hepatitis B virus (HBV)-related HCC with BCLC-B/C stage remain to be investigated., Methods: We retrospectively evaluated 126 patients with BCLC stage B/C HCC who underwent HR. These patients were divided into the antiviral group (Group A, n = 86) and the control group (Group B, n = 40). The operative indications and prognosis of 126 patients were analyzed., Results: The 1-year, 3-year, and 5-year disease-free survival (DFS) rates for Group A and Group B were 55.4%, 36.1%, 33.7% and 53.8%, 28.2%, 23.1%, respectively. The corresponding overall survival (OS) rates for the two groups were 89.2%, 61.4%, 45.8% and 82.1%, 48.7%, 33.3%, respectively. The DFS and OS for Group A were better than for Group B (p = 0.013, and p = 0.038, respectively). Antiviral therapy was an independent protective factor of late tumor recurrence [hazard ratio (HR) = 0.391, 95% confidence interval (CI): 0.190-0.806, p = 0.011] but not of early tumor recurrence., Conclusion: It is safe and feasible to perform HR combined with antiviral therapy for HBV-related HCC with BCLC stage B/C., (Copyright © 2016. Published by Elsevier Taiwan.)

Objective: To investigate the feasibility of contrast-enhanced computer tomography (CT) texture analysis in predicting early recurrence after transarterial chemoembolization (TACE) in patients with liver cancer. Methods: A retrospective analysis was performed for 47 patients with liver cancer confirmed by liver biopsy and digital subtraction angiography who underwent upper abdominal contrast-enhanced CT scan before TACE, and according to the presence or absence of focal recurrence within half a year, these patients were divided into early recurrence (ER) group and non-early recurrence (NER) group. The texture analysis was used to delineate tumor boundary layer by layer on the axial contrast-enhanced CT image before liver cancer surgery, and related parameters of tumor heterogeneity, including entropy, mean, non-uniformity, skewness, and kurtosis, were obtained. The independent samples t-test was used for comparison of texture parameters between the two groups. The receiver operating characteristic (ROC) curve was used for the analysis of entropy, mean, and non-uniformity, and the area under the ROC curve (ROC), optical cut-off value, sensitivity, and specificity were calculated to evaluate the efficiency of texture analysis in predicting early focal recurrence after TACE. Results: There were 20 patients in the ER group and 27 in the NER group. The ER group had a maximum major axis length of 88.2±36.3 mm and a maximum minor axis length of 41.4±21.4 mm, and the NER group had a maximum major axis length of 66.9±30.2 mm and a maximum minor axis length of 29.3±19.8 mm; the ER group had significantly higher maximum major and minor axis lengths than the NER group ( t = 4.89 and 4.62, P < 0.001). The ER group had significantly higher entropy and non-uniformity values than the NER group, and there were no significant differences in skewness and kurtosis between the two groups. Entropy, non-uniformity, and mean had high efficiency in predicting early recurrence after TACE, and the optimal cut-off value of entropy was 4.135. Conclusion: Volumetric texture analysis of contrast-enhanced CT images before liver cancer surgery has a high value in predicting early recurrence after TACE.

miRNAs are involved in substantial biological passways, including tumorigenesis, cancer development and progression. Angiogenesis plays a vital role in the progression of hepatocellular carcinoma (HCC), and VEGF is closely associated with the angiogenesis. However, the molecular mechanism of miRNAs in regulation tumorigenesis of HCC remains to be investigated. In the present research, we confirmed that miR-338-3p was suppressed both in HCC tissues and HCC cell lines. Then the tube formation, transwell and Chorioallantoic membrane (CAM) assay were carried out, such indicated that down-regulation of miR-338-3p can sharply increased, while up-regulation drastically suppressed angiogenesis of HCC cells in vitro. Moreover, MACC1 is predicted to be a target of miR-338-3p and we checked the prediction through luciferase assay. And then, our research showed that negative correlation existed between miR-338-3p and MACC1, β-catenin and VEGF that has been reported participated in cancer behavior in HCC cell lines. Subsequently, our assays illustrated that suppression miR-338-3p can up-regulate MACC1, β-catenin and VEGF expression of HCC cells. In conclusion, our research discovered that miR-338-3p can contribute to HCC angiogenesis by targeting MACC1, β-catenin and VEGF., (Copyright © 2016. Published by Elsevier Masson SAS.)

Aim: To investigate the effect of a sirolimus-based immunosuppressive protocol on tumor recurrence and survival after liver transplantation (LT) in hepatocellular carcinoma (HCC) patients., Methods: We retrospectively analyzed 142 HCC patients who underwent LT in our hospital from January 2006 to January 2012. The patients were divided into the sirolimus (SRL) group (62 cases) and non-sirolimus (control) group (80 cases). Disease-free survival (DFS) and tumor-bearing survival after tumor recurrence were compared using the Kaplan-Meier method., Results: No significant difference in DFS was observed between the two groups. Furthermore, DFS showed no significant differences in the subgroups of patients who met the Milan criteria, exceeded the Milan criteria but met the University of California, San Francisco (UCSF) criteria, or exceeded the UCSF criteria between the two groups. In the control group, 21 patients who were administered SRL after tumor recurrence had a median tumor-bearing survival time of 12months (3-34months), while 14 patients who did not experience a change in their immunosuppressive protocol after tumor recurrence had a median tumor-bearing survival time of 8months (6-22months). There was a significant difference in the tumor-bearing survival time between these patients (P=0.039)., Conclusions: Not all HCC patients benefited from the sirolimus-based immunosuppressive protocol after LT. However, sirolimus may prolong the survival time of patients after tumor recurrence., (Copyright © 2016. Published by Elsevier Masson SAS.)

Background: Elevated plasma fibrinogen is associated with tumour progression and poor outcomes in several cancers. The present study investigated the prognostic value of preoperative fibrinogen in hepatocellular carcinoma (HCC) patients after liver transplantation (LT)., Material and Methods: We analyzed the preoperative plasma fibrinogen levels of 41 patients who underwent LT for HCC. The cut-off value for elevated level of fibrinogen was determined by using a receiver operating characteristic (ROC) curve analysis. Cox regression analysis was performed to analyze the relationship between elevated fibrinogen level and HCC recurrence. The disease-free survival (DFS) and overall survival (OS) rate after transplantation were calculated by Kaplan-Meier method and compared by log-rank test., Results: The fibrinogen levels were significantly higher in patients with tumor recurrence (3.31±0.98 g/L) compared with those in patients without recurrence (2.39±0.89 g/L) (P<0.01). A cut-off value for elevated fibrinogen level of 2.675 g/L was defined. Cox regression analysis showed that the relative risk for tumor recurrence increased by 6.871 times for patients with elevated fibrinogen. Eleven patients in the elevated fibrinogen group (21 cases) developed recurrence, while only 2 in the normal fibrinogen group (20 cases) developed recurrence. There were significant differences in DFS and OS between the elevated fibrinogen group and normal fibrinogen group (5-year DFS and OS of 44.0% and 42.9% vs. 89.2% and 80.0%, respectively, P<0.05). Vascular invasion and fibrinogen level ≥2.675 g/L were the independent prognostic predictors of tumor recurrence and poor outcome., Conclusions: Pretransplant elevated fibrinogen levels are associated with tumor recurrence and poor prognosis in hepatocellular carcinoma patients after liver transplantation.

Background: Hepatitis B virus (HBV)-related end-stage liver disease is the leading indication for liver transplantation in China, but long-term results of liver transplantation in patients aged over 60 years are not clear. The present study was to reveal the natural history of liver recipients with hepatitis B older than 60 years., Methods: The recipients who had received liver transplantation between December 2003 and December 2005 were divided into two groups: those equal or older than 60 years (older group, n=60) and those younger than 60 years (younger group, n=305). Risk factors for poor long-term outcome in patients aged over 60 years were also analyzed., Results: Except for age and preexisting chronic disease (P<0.05), no significant differences were observed in perioperative characteristics between the two groups. There was also no significant difference in HBV and hepatocellular carcinoma recurrence (P>0.05). The actuarial 1-, 3-, 5- and 8-year survival rates were 81.6%, 71.6%, 66.7% and 63.3% respectively for the older group vs 84.9%, 77.7%, 70.8% and 65.6% for the younger group (P>0.05). Multivariate analyses showed that pre-liver transplant renal insufficiency was a risk factor for poor outcome in the older group (odds ratio=3.615, P=0.014)., Conclusions: Liver transplantation is safe and feasible for patients with HBV-related end-stage liver disease aged over 60 years. Older patients with renal insufficiency should undergo transplantation earlier than younger patients.

Cancer-associated fibroblasts (CAFs) are reported to support tumorigenesis by stimulating angiogenesis, cancer cell proliferation, and invasion in most solid tumors. However, the roles of CAFs in the liver cancer microenvironment have not been thoroughly studied. In our previous study, we successfully isolated CAFs from hepatocellular carcinoma (HCC) (H-CAFs) and proved that H-CAFs suppressed the activation of NK cells and thereby created favorable conditions for HCC progression. In our present study, we found that the proliferation of MHCC97L and Hep3B cells was significantly promoted by treatment with conditioned medium from H-CAFs. Pathological analysis also revealed that H-CAFs increased the proportion of Ki-67 (+) malignant cells and prevented them from undergoing necrosis. Moreover, the concentration of hepatocyte growth factor (HGF) cytokine in the conditioned medium of H-CAFs was higher than conditioned medium from normal skin fibroblasts (NSFs). Anti-HGF significantly reduced the proliferation-promoting capability of H-CAFs. In addition, we found that the abundance of H-CAFs correlated positively with tumor size. These results indicate that H-CAFs are an important factor for promoting the growth of HCC in vitro and in vivo, and that HGF plays a key role in HCC proliferation induced by H-CAFs.

Background: Up-regulation of Human Leukocyte Antigen-G (HLA-G) expression is thought to contribute to the escape in immune surveillance by suppressing natural killer (NK) cell function. However, little is known about the expression of HLA-G in hepatocellular carcinoma (HCC) and its relationship to NK cell-mediated cytotoxicity. In this study, we aimed to investigate the expression of HLA-G in human HCC cell lines and determine whether its expression was related to inhibition of NK cell cytolysis., Materials and Methods: The levels of HLA-G gene expression in human HCC cell lines were assessed by indirect immunofluorescence analysis (IFA), Real time RT-PCR and Western Blot. Vectors containing small interfering RNA (siRNA) specifically targeting the HLA-G gene were constructed and applied to diminish HLA-G expression. The cells were examined by flow cytometry and fluorescent microscope 24 h after transfection as well as 2-3 weeks after G418 selection. The steady-state levels of HLA-G mRNA and protein were then checked by real time RT-PCR and Western Blot analysis, respectively. A nonradioactive cytotoxicity assay was used to evaluate the effect of HLA-G on NK-mediated cytotoxicity against the siRNA treated cells., Results: Both HLA-G mRNA and protein can be detected in human HCC cell lines. Levels of HLA-G mRNA and protein were diminished 88.73% and 75.91% respectively by targeting siRNA. In the stable HLA-G gene knock-down cell lines, a significant increase in NK cell-mediated lysis occurred., Conclusions: Abnormal expression of HLA-G in HCC cell lines plays an important role in protecting against NK cell attack. The significant correlation between HLA-G expression and NK cell lysis implies that the abnormal expression of HLA-G might contribute to the mechanism of escape from host immune surveillance in HCC.

NADPH oxidase DUOX1, DUOX2, and NOX4 have recently been gained considerable concerns, owing to the fact that they involve in reactive oxygen species-induced genetic and epigenetic alternations of human carcinogenesis and serve as biomarkers in several cancers. Whether they predict survival in hepatocellular carcinoma (HCC) is still uncertain. Here, we detected the expressions of DUOX1, DUOX2, and NOX4 in one normal liver cell line, seven HCC cell lines, 30 non-cirrhotic normal liver tissues, and 107 paired HCC tissues using reverse transcription-polymerase chain reaction. The correlations of genes expression with prognoses were analyzed. DUOX1 was expressed at high levels in MHCC-97H and MHCC-97L, but at low levels in Bel-7402. In contrast to low expression level at SMMC-7721, DUOX2 was expressed at considerably high levels in MHCC-97H and MHCC-97L. The transcript of NOX4 was only detected in SMMC-7721. All the 30 normal liver tissues failed to express the three candidate markers. Compared with adjacent non-neoplastic tissues, DUOX1, DUOX2, and NOX4 were expressed at higher frequencies in tumor specimens. Both univariate and multivariate analyses revealed that elevated expression of DUOX1 or DUOX2 predicted poorer recurrence-free survival and overall survival. No such significance trend regarding NOX4 predictive value in survival, however, was seen in univariate analysis. These results suggested DUOX1 and DUOX2, but not NOX4, could predict HCC prognoses after hepatectomy.

Objective: To analyze the negative impact of preoperative neutrophil-lymphocyte ratio (NLR) on the tumor recurrence of hepatocellular carcinoma (HCC) after orthotopic liver transplantation., Methods: The clinical data of HBV (hepatitis B virus)-associated HCC patients undergoing liver transplantation were retrospectively analyzed. Their clinical and pathological risk factors for tumor-free survival were evaluated by univariate analysis. The analysis of Cox multiple regression was performed to determine the parameters of predicting the HCC recurrence. NLR ≥ 2.5 was considered to be elevated., Results: A total of 76 patients were identified. Among them, 37 had an elevated NLR. The 1, 3 and 5-year tumor-free survival rates were 69.2%, 52.7% and 50.9% respectively. The disease-free survival for patients with high NLR was significantly worse than that for those with normal NLR (1, 3, and 5 year survivals at 56.3%, 37.6% and 37.6% vs 81.1%, 66.9% and 63.3% respectively; P = 0.011). Univariate analysis of factors revealed that tumor size > 5 cm, tumor number > 3, vascular invasion, serum α-fetoprotein level ≥ 400 µg/L and NLR ≥ 2.5 were preoperative predictors of disease-free survival. Cox regression analysis showed that the presence of vascular invasion, tumor number > 3 and NLR ≥ 2.5 were independent prognostic factors of worse disease-free survival., Conclusion: An elevated NLR significantly increases the risk for tumor recurrence in HCC patients undergoing liver transplantation.

Objective: To investigate the relationship between hepatocellular carcinoma (HCC) recurrence and hepatitis B virus (HBV) recurrence., Methods: The clinical data of 340 patients underwent liver transplantation due to HBV related end-stage liver disease and received long-term follow up in our hospital from Jan 2004 to Dec 2008 were retrospectively analyzed. All patients received nucleoside analogues therapy formally before entering into the waiting list and nucleoside analogues combined low-dose HBIG therapy during and after transplantation. Patients were regularly followed up at the outpatient, monitoring the HBV recurrence and survival. Multivariate Cox regression analysis was used to evaluate the risk factors for hepatitis recurrence., Results: 33 patients suffered from HBV recurrence post transplantation. The 1-, 3- and 5- year recurrence rates were 7.0%, 10% and 13% respectively. The median HBV recurrence time was 5 months (1-21 months). COX regression analysis revealed that risk factors for HBV recurrence were HCC (HR = 2.98; 95% CI 1.08-8.25; P < 0.05) and pre-transplantation HBV-DNA load over 5 log10 copies/ml (HR = 3.99; 95% CI 1.85-8.62; P < 0.01). Further stratified analysis showed that patients who suffered from carcinoma recurrence had a higher incidence of HBV recurrence than those who did not, which were 27.9% and 8.7% (HR = 4.58;95% CI 1.88-11.12; P < 0.01) respectively. 12 patients suffered from both HCC and HBV recurrence. Spearman correlation analysis demonstrated a strong correlation between HBV and HCC recurrence times (r = 0.583, P < 0.05)., Conclusions: Post transplantation HCC recurrence is a risk factor for HBV recurrence.

Recently, Wnt inhibitory factor-1 (WIF-1) was found to be epigenetically inactivated in several solid tumors, but the biological and clinical relevance of WIF-1 methylation and expression status in hepatocellular carcinoma (HCC) are still unclear. In the present study, reverse transcription polymerase chain reaction (PCR) and methylation-specific PCR were used to examine the WIF-1 expression and methylation in HCC cell lines. In addition, methylation and expression status of WIF-1 in 105 HCC cases were correlated with clinicopathological parameters and prognosis after tumor resection. WIF-1 was expressed in one HCC cell line and L02, both of which were not methylated in promoter region. DNA hypermethylation of WIF-1 promoter was identified in the other four HCC cell lines without WIF-1 expression. In neoplastic and non-neoplastic tissue samples, the rates of WIF-1 methylation were 61.9% and 37.1% (P = 0.001), respectively. WIF-1 was significantly downregulated in neoplastic tissues at messenger ribonucleic acid (mRNA) level, as compared to adjacent non-neoplastic tissues (P = 0.006). A significant inverse association was observed between WIF-1 methylation of and WIF-1 expression (P  0.017, R = -0.232). Methylation of WIF-1 was not associated with patient survival. In contrast, patients whose tumors exhibited negative WIF-1 mRNA expression had lower rates of overall survival. These findings suggested that aberrant methylation of WIF-1 is a common event in hepatocarcinogenesis. In addition, expression, but not methylation, of WIF-1 is a predictor of good outcome in patients undergoing resection of HCC.

Aim: To analyze the relationship between hepatitis B virus (HBV) and hepatocellular carcinoma (HCC) recurrence in orthotopic liver transplantation (OLT) patients., Methods: 340 HBV patients with OLT were included in the study; among them were 148 patients with HBV-associated HCC., Results: HCC recurrence rates at 1, 3 and 5 years were 21, 29, and 46%, respectively. Exceeding Milan criteria (hazard ratio, HR = 12.35; 95% confidence interval, CI, 2.80-54.49; p = 0.001), HBV DNA level >5 log(10) copies/ml before transplant (HR = 2.45; 95% CI 1.10-5.45; p = 0.03) and HBV recurrence (HR = 2.27; 95% CI 1.10-4.75; p = 0.03) were significant independent predictors of HCC recurrence. HBV DNA >5 log(10) copies/ml before transplant (HR = 8.65; 95% CI 3.40-21.98; p < 0.001) and concomitance with HCC (HR = 2.79; 95% CI 1.33-5.87; p = 0.007) were predictors of HBV recurrence. Further stratified analysis showed that HBV recurrence was more prevalent in the HCC recurrence group (HR = 4.58; 95% CI 1.88-11.12; p = 0.001)., Conclusions: There is a close relationship between HBV and HCC recurrence after transplant. High HBV DNA levels before transplant are associated with HCC recurrence after transplant., (Copyright © 2011 S. Karger AG, Basel.)

Background: Neutrophil to lymphocyte ratio (NLR) has been proposed to predict prognosis of hepatocellular carcinoma (HCC). However, the cut-off values are empirical. We determined the optimal cut-off value to predict HCC recurrence after liver transplantation (LT) and further established a scoring model based on NLR., Methodology/principal Findings: We analyzed the outcome of 101 HBV-associated HCC patients undergoing LT. Preoperative risk factors for tumor recurrence were evaluated by univariate analysis. By using ROC analysis, NLR≥3 was considered elevated. The disease-free survival (DFS) and overall survival (OS) for patients with high NLR was significantly worse than that for patients with normal NLR (the 5-year DFS and OS of 28.5% and 19.5% vs. 64.9% and 61.8%, respectively; P<0.001). Univariate analysis revealed that tumor size >5 cm, tumor number >3, macrovascular invasion, AFP≥400 µg/L, NLR≥3, and HBV-DNA level >5 log10 copies/mL were preoperative predictors of DFS. Cox regression analysis showed macrovascular invasion, tumor number, and high NLR were independent prognostic factors. We then established a preoperative prognostic score based on multivariate analysis. Each factor was given a score of 1. Area under the ROC curve of the score was 0.781. All nine patients with score 3 developed recurrence within 6 months after LT. Of 71 patients without vascular invasion, three patients with both tumor number >3 and NLR≥3 developed recurrence within 14 months after LT while the 5-year DFS and OS for patients with a score of 0 or 1 were 68.1% and 62.8%, respectively., Conclusions/significance: Preoperative elevated NLR significantly increases the risk of recurrence in patients underwent LT for HCC. Patients with both NLR≥3 and tumor number >3 are not a good indication for LT. Our score model may aid in the selection of patients that would most benefit from transplantation for HCC.

Objective: To study the relationship between extrahepatic metastasis of primary hepatocellular carcinoma and circulative tumor cells in the blood of hepatoma patients., Methods: The immunomagnetic bead technique was employed to enrich and separate the hepatoma cells in the peripheral blood of preoperative and postoperative hepatoma patients. The relationship between postoperative extrahepatic metastasis and hepatoma cells in peripheral blood cancer cells were analyzed. The circulative tumor cells in the peripheral blood of hepatoma patients were enriched and separated by immunomagnetic bead technique. They were identified as hepatoma cells by AFP immunohistochemistry. Among 30 cases of hepatoma patients, the positive rate of hepatoma cells in the peripheral blood of preoperation and postoperation were 53.3% and 83.3% respectively. There was difference significantly in positive cases before operation and after operation (P<0.05)., Conclusions: Extrahepatic metastasis of primary hepatocellular carcinoma is obviously correlated to the positive tumor cells and the concentration in the peripheral blood of preoperative patients.

Objective: To study the effects of Rapamycin (RPM) on angiogenesis and tumor progression in human hepatocellular carcinoma (HCC) implantation mice., Methods: Tumor tissues of HCC were implanted into the liver of nude mice. Then, nude mice were treated with RPM and cyclosporine A (CsA). Real-time PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF). Immunohistochemical stain and image analysis were used to detect the protein expression of VEGF and proliferating cell nuclear antigen (PCNA) and microvessel density (MVD) was counted by endothelial cells immunostained by anti-CD34 antibody. The concentration of VEGF in the peripheral blood was detected by ELISA., Results: (1) The tumor weights were (372 +/- 35) mg, (769 +/- 39) mg and (751 +/- 42) mg in RPM, CsA and control group respectively. The tumor weight was significantly decreased in RPM group and no difference in CsA group compared with control group. (2) The expression of VEGF mRNA, VEGF and PCNA protein in tumor tissues and concentration of VEGF in the peripheral blood were remarkably down-regulated in RPM group compared with control group (P < 0.05) and were not remarkably different in CsA group from in control (P > 0.05).(3) Comparing with the control, the tumor MVD was remarkably decreased in RPM group (P < 0.05), and no difference in CsA group (P > 0.05)., Conclusion: RPM can inhibit angiogenesis and tumor progression of HCC by down-regulated the gene and protein expression of VEGF and inhibited the growth of tumor.

Objective: To screen the serum proteins associated with the metastasis of hepatocellular carcinoma (HCC) using a comparative proteomic approach., Methods: The serum samples of HCC patients with the same disease background were divided into metastatic (n=20) and non-metastatic (n=20) groups. The proteins extracted from the patients and 20 normal subjects, after depletion of the highly abundant proteins, underwent two-dimensional gel electrophoresis (2-DE). Comparative analyses of the 2-DE protein patterns between the 3 groups were conducted using a computerized image analysis system. The proteins with statistically significant differential expression between the metastatic and non-metastatic patients were identified by mass spectrometry. Western blotting was performed to examine the differential expression of the candidate proteins., Results: Four protein spots were identified by mass spectrometry among the 12 differentially expressed protein spots in the serum samples of HCC patients with intrahepatic metastasis, and confirmed by searching in MASCOT database. Of the 4 proteins, cytokeratin 9 (CK9) was up-regulated by 2 folds, and inter-alpha (globulin) inhibitor H4, complement factor H-related protein 1 precursor (FHR-1), and apolipoprotein E were down-regulated by 2 folds. CK9 was found to be specifically over-expressed in the metastatic group in comparison with the non-metastatic group, as confirmed by Western blotting., Conclusion: The metastasis of HCC might be correlated to the specific variation of protein expression profiles. The overexpression of CK9 may play a crucial role in HCC metastasis, and can be used as a potential serum marker for predicting HCC metastasis.

Objective: The present study aimed to evaluate the long-term outcomes and prognostic factors of elderly patients with hepatocellular carcinoma (HCC) undergoing hepatectomy., Material and Methods: From January 1983 to December 2006, 2,283 patients with HCC received hepatectomy in Sun Yat-sen University Cancer Center. The clinicopathological data and treatment outcomes of 67 elderly HCC patients (elderly group, > or =70 years of age) and 268 patients (control group, <70 years of age) who were selected randomly from the 2216 younger patients were compared retrospectively., Results: The elderly HCC patients had lower hepatitis B surface antigen-positive rate (P < 0.001), lower rate of marked alpha-fetoprotein elevation (P = 0.004), higher infection rate of hepatitis C virus (P = 0.010), more preoperative comorbidities (P < 0.001), higher rate of tumor encapsulation (P = 0.040), and better overall survival rate (P = 0.017); whereas there were no significant differences between these two groups in other factors, including gender ratio, liver function, accompanying cirrhosis, pathological tumor-node-metastasis (pTNM) staging, satellite nodules, vascular invasion, tumor rupture, resection margin, intraoperative blood loss, incidence of postoperative complications, hospital mortality, and disease-free survival rate. Multivariate analysis showed that pTNM staging was an independent prognostic factor of long-term survival in elderly patients with HCC., Conclusion: HCC in the elderly was less HBV-associated, less advanced, and less aggressive. Hepatectomy for selected elderly patients with HCC possibly have a better curative effect compared with younger patients. For the elderly patients without preoperative comorbidities or with controlled comorbidities, hepatectomy is a safe and effective treatment. pTNM staging is the only independent predictor of postoperative overall survival in elderly HCC patients.

Objective: To investigate the manifestation and diagnostic value of multislice spiral CT (MSCT) and MRI imaging in detection of tumor recurrence after liver transplantation for hepatocellular carcinoma (HCC)., Methods: The clinical data of 161 consecutive HCC patients who underwent orthotopic liver transplantation were retrospectively reviewed. Twenty-nine HCC patients were classified by pTNM according to the "Pittsburgh criteria". MSCT and MRI findings of tumor recurrence after liver transplantation were evaluated retrospectively in 29 stage II-IVb HCC patients. The recurrence site and relapse interval between liver transplantation and recurrence were analyzed., Results: Lung tumor recurrence were found in 21 cases, presented as cotton-like lesions in a diameter of 2 - 3 cm, with a clear margin and homogeneous density. Pleural tumor recurrence was detected in 4 cases. Liver tumor recurrence were found in 9 cases, which can be divided into four subtypes: multinodular in 4 cases, diffuse lesion in 2 cases, huge mass in 2 cases, and uninodular in 1 case. Two cases showed tumor thrombus in the inferior vena cava and portal vein. Lymph node tumor recurrence was found in 9 cases, presented as multiple nodules at hepatic hilum, lesser peritoneal sac, posterior mediastinum, retroperitoneum, or around pancreatic head, and accompanied with merging and necrosis in one case. Bone tumor recurrence were found as osteolytic destruction in 4 cases, and accompanied with adjacent soft-tissue mass in 2 cases. The recurrence sites of the 29 cases were as following: lung (21 cases, 72.4%), liver (9 cases, 31.0%), lymph nodes (9 cases, 31.0%), bone (4 cases, 13.8%) and other sites (3 cases, 10.3%). Lung tumor recurrence was found in all the 10 stage IVb patients with tumor recurrence after liver transplantation, significantly more frequent than that in stage IVa patients (P = 0.023). After liver transplantation, all 25 patients with stage III approximately IVb HCC developed recurrence within one year, but in the 4 cases with stage II HCC at one year later (P = 0.009)., Conclusion: The results of our study show that in hepatocellular carcinoma patients after liver transplantation, the lung and pleura are the most frequent site of recurrence, followed by liver, lymph node and bone as the second and third sites. The Stage IVb hepatocellular carcinoma should be regarded as a contradiction for liver transplantation due to rapid recurrence. Tumor recurrence occurs later in stage II HCC than in stage III approximately IVb patients. MSCT and MRI are of significant importance in diagnosis and formulating operation plan in HCC patients with recurrence after liver transplantation.

Objective: To study the effects of sirolimus (SRL) on the growth of transplanted human hepatocellular carcinoma (HCC) in nude mice., Methods: HepG2 cells were Implanted into the liver of nude mice. The implanted mice were then treated with SRL and tacrolimus (FK506). The expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) was detected by immunohistology, microvessel density (MVD) was counted by immunostaining with anti-CD34 antibody for endothelial cells. Tumor apoptosis was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay., Results: The tumor weight was (352+/-38) mg, (683+/-53) mg and (675+/-45) mg in SRL, FK506 and control group respectively. The tumor weight was significantly decreased in SRL group (P < 0.01), and there was no difference between FK506 group and control group. The expression of VEGF and PCNA protein was remarkably down-regulated in SRL group compared to control group (P < 0.05), and it was not significantly different between FK506 group and control group (P > 0.05). Compared to the control group, MVD was significanly decreased in SRL group, and the apoptosis index of tumor cell was significantly higher in SRL group (P < 0.01)., Conclusion: SRL inhibits transplanted HCC tumor growth by reducing tumor angiogenesis, inhibiting tumor proliferation and inducing tumor apoptosis.

Objective: To investigate the effect of rat mesenchymal stem cells (MSCs) transfected with different oncogenes differentiate into hepatocellular carcinoma (HCC) in vitro., Methods: MSCs, transfected with different oncogenes c-myc, K-ras, c-myc and K-ras and amplified in vitro, were infused into rats via vena portae. The recipient rats were divided into the hepatic impairment group, which were fed with tetrachloromethane and the healthy control group. At day 7, 14, 21, 28 and 42 following grafting, the complete livers were obtained and examined using fluorescence detection, conventional pathology and immunohistochemistry detection of GFP, c-kit and AFP to study the colonization and distribution of stem cells in rat liver., Results: No immunological rejection occurred after grafting of allogenic MSCs. The infused MSCs colonized in the recipient rat liver. Liver tumors were present in 6 rats grafted with MSCs that were transfected with K-ras, K-ras and c-myc, and the protein expression of AFP was detected using immunocytochemistry at day 7. Rats grafted with MSCs that were transfected with c-myc gene had no obvious tuberosity or tumor. Small oval cells were found microscopically in the periphery of vena portae, and immunohistochemistry staining of AFP was negative. Immunohistochemical staining of c-kit was positive in all livers of rats that were transfected with MSCs., Conclusions: Hepatocellular carcinoma may derive from genetically mutated MSCs.

Background: Rapamycin, isolated from Streptomyces hygroscopicus, is recently reported to have immunosuppressant and anti-tumor effects on a large variety of cancers. To date, no detailed data are available about the effects of rapamycin on hepatocellular carcinoma cells., Objective: In this study, the anti-proliferation effects of rapamycin on hepatocellular carcinoma cells BEL-7402 and HepG-2 in vitro were studied., Methods: Cell viability was assessed by MTT assay and [3H]-thymidine uptake, cell apoptosis was observed by Hoechst 33258 staining and flow cytometry (FCM). The variation of caspase-3 and apoptotic related genes was assayed by Western blotting, cell mitochondrial membrane potential was also investigated by using standard methods., Results: Rapamycin could inhibit the growth of hepatocellular carcinoma cells and cause apoptosis significantly; the suppression was both in time- and dose-dependent manner, marked morphological changes of cell apoptosis were observed very clearly by Hoechst 33258 staining. Rapamycin exhibits induction apoptosis by activation of caspase-3 and disruption of the mitochondrial membrane potential on hepatocellular carcinoma cells in vitro. Western blotting analysis demonstrated that anti-apoptotic protein Bcl-2 was down-regulated while pro-apoptotic protein Bcl-xl up-regulated remarkably in a time-dependent manner when apoptosis occurred., Conclusion: Rapamycin has significant anti-proliferation effect by induction of apoptosis via activation of caspase-3 and disruption of mitochondrial membrane potential, as well as by down-regulation of anti-apoptotic protein Bcl-2 and up-regulation of pro-apoptotic protein Bcl-xl on hepatocellular carcinoma cells. The data provide a potential mechanism for rapamycin-induced apoptosis in hepatocellular carcinoma cells, suggesting that rapamycin may serve as both an effective adjunctive reagent for the treatment of residual cancer cells and immunosuppressant after liver transplantation of hepatocellular carcinoma, and that in vivo anti-cancer effects as well as its potential clinical effectiveness need further investigation.

Aim: To develop a conditionally replicative gene-viral vector system called CNHK500-p53, which contains dual promoters within the E1 region, and combines the advantages of oncolytic virus and gene therapies for hepatocellular carcinoma (HCC)., Methods: CNHK500-p53 was constructed by using human telomerase reverse transcriptase (hTERT) promoter to drive adenovirus E1a gene and hypoxia response element (HRE) promoter to drive adenovirus E1b gene. p53 gene expressing cassette was inserted into the genome of replicative virus. Viral replication experiments, cytopathic effect (CPE) and methyl thiazolyl tetrazolium (MTT) assay were performed to test the selective replication and oncolytic efficacy of CNHK500-p53., Results: Immunohistochemistry verified that infection with CNHK500-p53 was associated with selective replication of adenovirus and production of p53 protein in telomerase-positive and hypoxia-inducible factor-dependent HCC cells. p53 protein secreted from HepG2, infected with CNHK500-p53 was significantly higher than that infected with nonreplicative adenovirus Ad-p53 in vitro (388 +/- 34.6 microg/L vs 76.3 +/- 13.17 microg/L). Viral replication experiments showed that replication of CNHK500-p53 and CNHK500 or WtAd5, was much stronger than that of Ad-p53 in tested HCC cell lines. CPE and MTT assay indicated that CNHK500-p53 selectively replicated in and killed HCC cells while leaving normal cells unaffected., Conclusion: A more efficient gene-viral system is developed by combining selective oncolysis with exogenous expression of p53 against HCC cells.

Background and Aim: Ponicidin is recently reported to have anti-tumour effects on a large variety of cancers. The present study was undertaken to investigate the anti-proliferation effects of ponicidin on hepatocellular carcinoma cells and its mechanism., Methods: Two hepatocellular carcinoma cell lines, QGY-7701 and HepG-2 cells, were used. Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptosis was assessed by flow cytometry and deoxyribonucleic acid fragmentation analysis. Cell morphology was observed by Hoechst 33258 staining. Reverse transcriptase-polymerase chain reaction and Western blot analysis were used to detect Survivin as well as Bax and Bcl-2 expressions., Results: Ponicidin could inhibit the growth of QGY-7701 and HepG-2 cells significantly by induction of apoptosis. Marked morphological changes of apoptosis were observed clearly. Both Survivin and Bcl-2 expressions were down-regulated remarkably while Bax expression up-regulated when apoptosis occurred., Conclusions: Ponicidin has significant anti-proliferation effects by inducing apoptosis on hepatocellular carcinoma cells in vitro, down regulation of Survivin and Bcl-2 as well as upregualation of Bax expressions may be the important apoptotic inducing mechanisms.

Background & Objective: Rapamycin, isolated from Streptomyces hygroscopicus, is recently reported to have immunosuppressive and antitumor effects on a large variety of cancers. This study was to investigate the role of Caspase-3 in rapamycin-induced apoptosis of hepatocellular carcinoma BEL-7402 cells., Methods: BEL-7402 cells were treated with different concentrations (5, 10, 20, 30, 40, 50 nmol/L) of rapamycin. Cell viability was detected by MTT assay; cell apoptosis was observed by flow cytometry (FCM) and Hoechst 33258 staining. The activity of Caspase-3 was determined by Caspase colorimetric assay kit and Western blot., Results: Rapamycin inhibited the growth of BEL-7402 cells and induced apoptosis significantly in time- and dose-dependent manners. Marked morphologic changes of cell apoptosis, such as chromatin condensation and nuclear fragmentation, were observed clearly at 48 h after exposion to rapamycin; Caspase-3 was activated by the loss of Caspase-3 proenzyme (32-ku) and its 20-ku subunit appeared at 24 h after incubation. Caspase-3 inhibitor z-DEVD-FMK could block the apoptosis induced by rapamycin., Conclusions: Rapamycin can inhibit growth and induce apoptosis of BEL-7402 cells. The activation of Caspase-3 may play an important role in cell apoptosis.

Objective: To investigate the change of Bcl-2 expression and telomerase during the apoptosis induced by oridonin on human hepatocelluar carcinoma BEL7402 cells., Method: BEL-7402 cells in culture medium were given 8,16,24,32 micromol x L(-1) different concentrations of oridonin. The cell apoptotic rate was detected by flow cytometry (FCM), morphology of cell apoptosis was observed by Hoechst 3325 staining. Bcl-2 and Bax expressions were detected by Western blotting. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR enzyme-linked immunosorbent assay (ELISA) were used to detect hTERT mRNA expression and telomerase activity., Result: Oridonin induced BEL-7402 cells apoptosis significantly, and the apoptosis rate was both in time-and dose-dependent manner. Marked morphological changes of cell apoptosis were observed very clearly by Hoechst 33258 staining after the cells exposed to oridonin for 60 hours; Western blotting showed that Bcl-2 expression was down-regulated and Bax expression up-regulated concurrently along with the apoptotic process, and the expression of hTERT mRNA as well as activity of telomerase were decreased concurrently., Conclusion: Oridonin could decrease the expression of hTERT mRNA and telomerase activity as well as down-regulation of Bcl-2 and up-regulation of Bax expression during the apoptosis of BEL-7402 cells.

Objective: To investigate the effect of adjuvant individualized chemotherapy in prevention of tumor recurrence and improvement of patient survival after liver transplantation for hepatocellular carcinoma (HCC)., Methods: 21 HCC cases received orthotopic liver transplantation and treated with adjuvant individualized chemotherapy based on ATP tumor chemosensitivity assay (ex vivo) between April 2001 and January 2003 were retrospective reviewed, compared with 52 cases received orthotopic liver transplantation only. The cumulative and tumor-free survivals were compared between 2 groups., Results: The 1, 2 years overall survival rates were 92.31%, 73.85% for the individualized chemotherapy group and 92.06%, 63.93% for the non-chemotherapy group, the difference was not statistically significant. The 6, 12, 18, 24 months tumor-free survival rates were 90.00%, 80.00%, 80.00%, 60.00% and 67.31%, 51.92%, 40.03%, 37.81% respectively, the difference was statistically significant (P <0.05)., Conclusions: This study suggests that tumor recurrence decreases and tumor-free survival increases by adjuvant individualized chemotherapy after liver transplantation for HCC. The individualized protocol based on ATP-TCA may be effective for patients with HCC after liver transplantation.

Background & Objective: Most cases of hepatocellular carcinoma (HCC) are unresectable even without extra-hepatic metastasis. In addition, about 80%-90% patients in China are accompanied with cirrhosis. Liver transplantation is probably more proper for these patients than other treatments. This study was to explore the value of liver transplantation for treatment of advanced HCC without extra-hepatic metastasis, and to summarize the experience of liver transplantation performed in Cancer Center of Sun Yat-sen University., Methods: Ten patients with advanced HCC without extra-hepatic metastasis who underwent liver transplantation in Sun Yat-sen University Cancer Center from Sept. 2003 to Apr. 2004 were followed up to May 2004, and their clinical records were reviewed retrospectively., Results: The mortality within 1 month after operation was 0.0% for 10 cases of liver transplantation. One patient was alive for 7 month with tumor recurrence, and the remaining 9 patients were alive for 1-6 months without recurrence. Alpha fetoprotein (AFP) decreased to normal range within 2 months in 2 cases with macro tumor thrombus in the trunk of portal vein, and the left branch of portal vein, respectively., Conclusion: Advanced HCC without extra-hepatic metastasis, even with macro tumor thrombus in portal vein, is still indicated for liver transplantation.

Objective: To evaluate the role of orthotopic liver transplantation (OLT) in treatment of hepatocellular carcinoma (HCC) and the selection of recipients., Methods: OLT was performed in 60 patients with HCC at Organ Transplantation Centre of the First Affiliated Hospital of Sun Yat-sen University between September 1993 and September 2002. Medical records were retrospectively analyzed with regard to the response to OLT and survival., Results: One-month, 1, 2, 3-year survival rate of 23 liver transplant performed from September 1993 to July 2002 were 73.9%, 60.9%, 43.5% and 29.0%, respectively. One-month, 1, 2-year survival rate of 37 liver transplant performed from August 2000 to September 2002 were 89.2%, 75.8% and 61.2%, respectively. One-month survival rate was significantly greater in the patients with a preoperative liver function of Child A or B than Child C (P < 0.05). One-month, 1, 2, 3-year survival rate of small HCC (single tumor 5 cm diameter, n = 41) were 84.2%, 76.6%, 65.6%, 65.6% and 82.9%, 63.1%, 46.7%, 37.4%, respectively. The median survival of large HCC was 18.0 months and mean survival of small HCC was 29.6 months, respectively. The recurrence rate of small HCC and large HCC were 15.8% and 27.7%, respectively. There was no significant difference between the cumulative survival of small HCC and large HCC. The quality of life of patients with long-term survival was good., Conclusions: HCC associated with cirrhosis can be effectively treated by OLT on condition that no extrahepatic spread and no vascular involvement. OLT is recommended for treatment of small HCC combined with liver cirrhosis, meanwhile, OLT performed in the partial large HCC still is reasonable at the present time.

Background/aims: Familial clustering of hepatitis B virus (HBV) infection is related to perinatal transmission, and is the main cause of familial-type hepatocellular carcinoma (HCC). The route of HBV transmission differs between the children and siblings of patients with HCC. This study examined the differences in HBV carrier rates and HCC-related mortality between two generations in HCC families., Methods: From 1992 to 1997, relatives of individuals with HCC were screened prospectively with ultrasonography, alpha-fetoprotein, liver biochemistry tests and viral markers. Total HCC-related deaths during a 9-year period were compared between the generations of index patients and their children., Results: The study included a total of 13676 relatives in two generations. More HCC-related deaths occurred in the index patient generation than in the child generation. Furthermore, children of female index patients had higher rates of liver cancer related mortality than children of male index patients. The same was true when the analysis was limited to male HBV carriers. The prevalence of HBsAg in the offspring of HBsAg positive mothers was 66% in the child generation and 72% in the index patient generation. These high prevalences indicated high maternal HBV replication status., Conclusions: Perinatal transmission and maternal viral load are important risk factors in hepatocarcinogenesis.

Background/aims: Technetium-99m methoxyisobutyl isonitrile (MIBI) has been shown to be useful in identifying several types of tumors, such as breast, lung and thyroid cancers. There are only a few reports in the literature regarding Tc-99m MIBI uptake in hepatocellular carcinoma, and the results are conflicting. The aim of this study was to investigate the relationship between Tc-99m MIBI accumulation in patients with hepatocellular carcinoma and the gene expression of P-glycoprotein-multidrug-resistance., Methodology: Twenty-two patients with hepatocellular carcinoma were enrolled in this study. Tc-99m MIBI imaging was performed 10 minutes after intravenous injection of 20 mCi Tc-99m MIBI. All patients had liver biopsy or surgery within 1 week of MIBI imaging. Immunohistochemical study of the biopsy or resected hepatocellular carcinoma specimens was performed using the anti-human P-glycoprotein antibody., Results: On Tc-99m MIBI imaging, 20 of 22 (90.9%) patients with hepatocellular carcinoma showed negative Tc-99m MIBI uptake in tumor lesions, whereas only 2 patients showed positive Tc-99m MIBI uptake in tumor lesions. P-glycoprotein expression was observed in 13 of 20 (65%) patients with negative Tc-99m MIBI uptake, but in the 2 patients who showed positive Tc-99m MIBI uptake in tumor lesions, P-glycoprotein expression was negative., Conclusions: Tc-99m MIBI SPECT is a useful noninvasive method for predicting the expression of P-glycoprotein-multidrug-resistance gene in hepatocellular carcinoma patients.

Background/aims: Hepatocellular carcinoma is one of the most common malignancies in Taiwan, a hepatitis B endemic area. In this study, we aimed to evaluate the characteristics of hepatocellular carcinoma in patients receiving hemodialysis, as this patient group is at high risk for malignancy., Methodology: From October 1991 to September 1997, thirteen patients receiving hemodialysis and diagnosed with hepatocellular carcinoma were enrolled in this retrospective study. Patients' clinical course, laboratory data, image study and treatment were evaluated., Results: Among these 13 patients, hepatitis B virus related in 6 and hepatitis C virus related in 7. There was no statistical significance in serum levels of asparate aminotransferase, alanine aminotransferase, bilirubin and alpha-fetoprotein between hepatitis B virus and hepatitis C virus related hepatocellular carcinoma. Hepatitis B virus related hepatocellular carcinoma had a shorter mean dialysis period (29.67 +/- 22.18 vs. 87.86 +/- 79.90 months, P = 0.25) and mean duration from beginning hemodialysis to diagnosis of hepatocellular carcinoma (17.16 +/- 26.94 vs. 76.08 +/- 65.69 months, P = 0.05), but there was no statistical significance. In the area of treatment, the survival curves of the treatment (hepatic resection and/or transcatheter arterial chemoembolization) group and supportive group were compared by log-rank test and there was no statistical significance for these two groups (P = 0.69)., Conclusions: Both hepatitis B virus and hepatitis C virus are equally important for hepatocellular carcinoma in hemodialysis patients in hepatitis B endemic Taiwan. The acquisition of hepatitis B virus or hepatitis C virus might be not related to hemodialysis. Periodic screening with ultrasonography and serum alpha-fetoprotein is necessary among hemodialysis patients with evidence of hepatitis B virus or hepatitis C virus infection.

Background/aims: Hepatocellular carcinoma is part of the natural history of liver cirrhosis. Gastrointestinal bleeding and hepatic failure are the leading causes of death in hepatocellular carcinoma patients. With gastrointestinal bleeding, variceal bleeding is the most prominent, and most variceal bleeding is of esophageal origin. Gastric varices bleeding is often a massive and severe bleeding episode. The role of gastric varices among patients with hepatocellular carcinoma remains to be clarified. In this study, we aimed to evaluate the prevalence, clinical significance and prediction of gastric varices in patients with hepatocellular carcinoma., Methodology: From 1998 to 2000, we reviewed 304 patients with hepatocellular carcinoma receiving upper gastrointestinal endoscopic examinations. Patients' clinical characteristics, physical findings, laboratory data, image studies, endoscopic examinations and treatment were reviewed., Results: Among 304 patients with HCC, twenty-one (6.9%) had gastric varices among 304 patients with hepatocellular carcinoma. The location of gastric varices were the posterior wall in 12 (57%), the lesser curvature in 1 (5%), the greater curvature in 4 (19%) and the fundus in 4 (19%). Three (14%) of these 21 patients with hepatocellular carcinoma and gastric varices had clinical evidence of bleeding. One of them died due to uncontrollable bleeding. Child-Pugh classification, hepatic encephalopathy, portal vein or splenic vein dilatation, ascites, splenomegaly, albumin level, prothrombin time and platelet count were significantly different between hepatocellular carcinoma patients with gastric varices and without gastric varices under the univariate analysis. Ascites (Odds ratio: 5.45; 95% confidence interval: 2.12-14.01) and portal vein or splenic vein dilatation (Odds ratio: 4.38; 95% confidence interval: 1.77-10.86) were the two most important predictors under the stepwise logistic regression analysis., Conclusions: The prevalence of gastric varices in patients with hepatocellular carcinoma is 6.9% and the risk of bleeding is low in this study. The Predictors of gastric varices among hepatocellular carcinoma are related to liver cirrhosis, Child-Pugh classification, hepatic encephalopathy, portal vein or splenic vein dilatation, ascites, splenomegaly, albumin level, prothrombin time and platelet count.

Background/aims: To study and review the clinical manifestations, microbiology, comorbidity, and diagnosis of liver abscess after transarterial chemoembolization for malignant hepatic tumor., Methodology: We retrospectively reviewed 1374 patients who underwent 2581 transarterial chemoembolization procedures due to malignant hepatic tumors over an 8-year period., Results: 7 patients had liver abscess after transarterial chemoembolization. The incidence was 0.27% (7/2581). Hepatocellular carcinoma was diagnosed in all 7 patients, whose liver function was classified as stage A by the Child-Pugh criteria. The clinical manifestations were intermittent fever, abdominal pain, and leukocystosis. All the patients had hyperechoic spots with reverberative shadows on sonograms or low attenuation areas with different Hounsfield units on computed tomography scan, which expressed the 100% incidence (7 of 7) of gas-forming abscesses. Percutaneous drainage or aspiration was done in 6 patients. One received laparotomy with local debridement due to suspicious organ rupture. The pus culture showed Gram-negative bacteria in all patients. Blood cultures were positive in only 3 of 7 patients (43%). No patients died of liver abscess after aspiration, drainage, or debridement of abscess combined with parenteral antibiotic treatment. Biliary tract diseases, found in 4 patients, were the most common comorbidity., Conclusions: Liver abscess after transarterial chemoembolization is a very rare complication, which usually develops in patients with biliary tract disease. Gram-negative bacteria are the main pathogens. The incidence of gas formation is higher after transarterial chemoembolization than in the general population. However, the prognosis is good after adequate clearance of pus and antibiotic treatment.

P-glycoprotein (Pgp) and multidrug resistance-associated protein (MRP) expressions as well as Tc-99m methoxisobutylisonitrile (MIBI) images were assessed in 25 patients hepatocellular carcinoma (HCC). Tc-99m MIBI imaging was performed 10 minutes after intravenous injection of 20 mCi Tc-99m MIBI. Using immunohistochemical staining, 60% of the HCC lesions showed positive for Pgp and 64% showed positive for MRP. In 3 patients with MIBI uptake, immunohistochemical study of tumor tissue showed no Pgp stained cells. Nevertheless, they were all positive for MRP. The result of Tc-99m MIBI imaging is more related to the expression of Pgp than MRP gene. It is possible that other membrane transporters as well as Pgp and MRP are involved in the efflux of Tc-99m MIBI.

Background/aims: Combined hepatocellular-cholangiocarcinoma is a rare subtype of primary liver cancer. To clarify the significance of this tumor, we conducted the present study to investigate the clinicopathological characteristics. As well, a comparison between this tumor and cholangiocarcinoma was made to determine whether these two types of liver cancers are closely related., Methodology: Seventeen cases of combined hepatocellular-cholangiocarcinoma and 104 cases of cholangiocarcinoma were included. Patients were evaluated on the basis of age, gender, viral hepatitis markers, tumor markers, laboratory data, indocyanine green test, tumor location, histological differentiation, lymph node involvement, and organ metastasis. In addition, the presenting symptoms at admission were also recorded. Data was analyzed using the chi 2 test and Student's t test and differences between the curves were tested using the two-tailed log-rank test., Results: Combined hepatocellular-cholangiocarcinoma was found to be more prevalent in males. In addition, the combined hepatocellular-cholangiocarcinoma patients had lower serum bilirubin, ALP levels (p < 0.05), and more poor differentiation (p < 0.05). Also, the presenting symptoms of combined hepatocellular-cholangiocarcinoma patients were different from those of cholangiocarcinoma patients with the former a lower incidence of fever, chills, jaundice and a higher incidence of fatigue and weakness., Conclusions: The clinicopathological features of combined hepatocellular-cholangiocarcinoma were different from those of cholangiocarcinoma with more advanced histological differentiation, more prevalent in males, and lower levels of serum bilirubin and ALP. These characteristic features may help us in diagnosing combined hepatocellular-cholangiocarcinoma in patients with suspected cholangiocarcinoma.

Background: Families of patients with hepatocellular carcinoma (HCC) carry a high risk of developing HCC. We determine the number of fatalities in relatives of HCC patients during an 8-year period to understand the risk and cause of HCC in relatives of patients with HCC., Methods: From 1992 to 1997, 15 410 relatives of HCC patients in three generations were screened prospectively for HCC by ultrasonography, alpha-fetoprotein, liver biochemistry and viral markers. By using national citizen identification numbers, we searched the total fatalities in relatives of HCC patients between 1992 and 1999 from the national mortality data bank. The results were compared among different viral infection groups., Results: Of the relatives studied, 37.8% were hepatitis B s antigen (HBsAg) positive (+), 4.3% were anti-hepatitis C virus (HCV) (+) and 1.7% were both HBsAg (+) and anti-HCV (+). A total of 399 fatalities, including 139 because of HCC (34.8%), 37 because of liver diseases (9.3%), 88 because of other cancers (22.1%) and 135 because of other diseases (33.8%), were found. Relatives who were HBsAg (+) or anti-HCV (+)showed a lower cumulative survival than did relatives who were negative for both HBsAg and anti-HCV. Relatives with dual infection of hepatitis B and C virus showed the highest mortality due to HCC or terminal liver diseases., Conclusions: Chronic viral infection rather than a hereditary factor is the main cause of a familial tendency for HCC. Dual infection of hepatitis B and C virus increases the risk of HCC or decompensated liver diseases.

Hepatocellular carcinoma (HCC) is one of the most common malignancies worldwide, especially in Asia. Gastrointestinal bleeding due to esophagogastric variceal hemorrhage is one of the leading causes of death in HCC patients. The aim of study was to determine whether clinical variables were predictive of the presence of large esophagogastric varices (EGV) before performing endoscopy. Three hundred and four HCC patients who received endoscopy were enrolled and studied retrospectively. Univariate and stepwise logistic regression analysis were used to evaluate associations between the presence of large EGV and patient characteristics. There were 248 patients with small or no EGV and 56 patients with large EGV. The optimal critical values determined by a receiver operating characteristic curve for platelet count and albumin level were 135,000/mm3 and 3.5 g/dl, respectively. Stepwise logistic regression analysis demonstrated that splenomegaly [odds ratio (OR): 9.72; confidence interval (CI): 3.75-25.17], portal vein thrombosis (OR: 2.73; CI: 1.50-4.97), low platelet count (<135,000/mm3) (OR: 3.78; CI: 2.07-6.90) and low albumin level (<3.5 g/dl) (OR: 3.44; CI: 1.73-6.82) were significant, independent predictors for large EGV. Large EGV also could be independently predicted by Child-Pugh classification, splenomegaly (OR: 4.93; CI: 1.87-13.01), or portal vein thrombosis (OR: 2.37; CI: 1.28-4.39) while excluding the non-cirrhotic patients. In conclusion, splenomegaly, low platelet count (<135,000/mm3), and low albumin level (<3.5 g/dl) are clinical predictors to stratify HCC patients at risk of developing large EGV. Besides factors related to liver cirrhosis, portal vein thrombosis is also an important predictor for HCC patients with large EGV.

In conclusion, pyogenic liver abscess in hepatocellular carcinoma is unusual. Most of the reported cases occurred after a treatment such as transcatheter arterial embolization or percutaneous ultrasound-guided ethanol injection. Salomonella very rarely causes pyogenic liver abscesses. Only 14 cases have been reported in the English literature since 1911. Salmonella liver abscess occurring within a primary neoplasm is even rarer. There were only two such cases described in patients with hepatocellular carcinoma before. The present case is the third one, but it may be the first case of obvious spontaneous gas-forming liver abscess caused by Salmonella within hepatocellular carcinoma.

Background and Aims: In recent years, gastric slow-wave dysrhythmias induced by transcatheter arterial chemoembolization (TACE) have been observed. Enhanced endogenous prostaglandin may be a possible mechanism for the myoelectrical changes. The aim of this study was to evaluate whether the gastric slow-wave dysrhythmias induced by TACE may be mediated by ketoprofen, a prostaglandin synthesis inhibitor., Methods: Twenty-three patients with hepatocellular carcinoma (HCC) admitted for TACE were enrolled. A follow-up TACE was scheduled to take place 2 months later. During the next admission for TACE, 50 mg of ketoprofen was given intramuscularly 12 h for 3 days, beginning 48 h before TACE, as premedication. Cutaneous electrogastrography (EGG) was performed before and within 24 h after TACE., Results: The results showed that the change in the fasting EGG parameters after TACE without premedication was not statistically significant. However, the postprandial EGG parameters, including the dominant frequency (DF); the percentages of DF in the normal, bradygastric and tachygastric range; along with the dominant frequency instability coefficient, deteriorated significantly after the procedure (P < 0.01). After the follow-up TACE with ketoprofen premedication, neither the fasting nor postprandial EGG parameters in the control group changed significantly., Conclusions: Gastric slow-wave dysrhythmias induced by TACE may be mediated by ketoprofen, a prostaglandin synthesis inhibitor, in HCC patients. However, the improvement in the gastric myoelectrical activity does not eliminate the degree of nausea/vomiting after TACE.

Background/aims: In Taiwan, most cases of hepatocellular carcinoma (HCC) are hepatitis B virus (HBV) or hepatitis C virus (HCV) related. The serum alpha-fetoprotein (AFP) level is an important factor in the diagnosis of HCC. There have been many studies discussing the role of AFP in diagnosing HBV-related HCC, but only few concerning HCV-related HCC. In this study, we aimed at analyzing the distribution of AFP levels in anti-HCV positive patients with and without HCC and evaluating the effectiveness of serum AFP levels in screening HCV-related HCC., Methodology: From 1993-1996, we collected the AFP data of 205 HCC patients retrospectively, who were anti-HCV positive. For comparison, 131 randomized anti-HCV positive patients without evidence of HCC served as the control group. We analyzed the AFP distribution in both groups over the following ranges: < or = 5 ng/ml, > 5-20 ng/ml, > 20-50 ng/ml, > 50-100 ng/ml, > 100-200 ng/ml and > 200-400 ng/ml, and > 400 ng/ml., Results: The distributions of AFP levels in anti-HCV positive patients with HCC were 13.2%, 21.5%, 11.2%, 4.9%, 4.4%, 7.3%, and 37.6%. The distributions in anti-HCV positive patients without evidence of HCC were 34.3%, 55.0%, 8.4%, 1.5%, 0.8%, 0%, 0%., Conclusions: We found the differences in AFP to be statistically significant between anti-HCV positive patients with and without HCC. A serum AFP level of more than 200 ng/ml highly indicates HCC. However, there is a large overlap between these 2 groups. Thus, in anti-HCV positive patients, AFP level is not a good single reference for diagnosis of HCC. Anti-HCV positive patients should be routinely screened for HCC by image studies along with serum AFP level.

Background: Transcatheter arterial chemoembolization (TACE) of the hepatic artery is frequently used in the treatment of inoperable hepatocellular carcinoma (HCC). TACE causes not only effective tumor tissue necrosis in patients with hepatoma but also adverse effects on extrahepatic abdominal organs. There are no published reports on the effect of TACE on the gastric myoelectric activity. In this study, using cutaneous electrogastrography (EGG), we evaluated the effect of TACE on gastric myoelectric activity in patients with HCC., Methods: A total of 27 patients (24 men and 3 women, aged 22 to 78 years) with hepatoma, admitted for TACE, were included in this study. Furthermore, 28 patients (24 men and 4 women, aged 26 to 75 years), admitted for diagnostic angiography of the liver, served as the control group. Cutaneous EGG was performed before and after TACE or angiography., Results: In the TACE group there were significant changes in dominant frequency (DF) and percentages of DF in the defined normal range, bradygastric range, and tachygastric range on post-meal EGG. On fasting EGG, only the dominant frequency and percentages of DF in the bradygastric range changed significantly. However, there was no correlation between the occurrence of nausea/vomiting and the degree of change in the EGG variables, during both fasting and postprandial states. In the control group there were no significant differences in EGG variables before and after angiography., Conclusions: TACE can affect gastric myoelectric activity in HCC patients. Nevertheless, the relationship between changes in myoelectric activity and the occurrence of gastrointestinal symptoms needs further investigation.

Between August 1983 and December 1991 at the Taichung Veterans General Hospital, Taiwan, 65 advanced hepatocellular carcinoma (HCC) patients with esophageal variceal bleeding received endoscopic injection sclerotherapy (EIS) and 60 such patients received conservative medical treatment without EIS. The rate of successful control of acute bleeding was 72.5% (27/40 patients) in the EIS group and 56.7% (34/60 patients) in the non-EIS group. The rebleeding rate was lower in the EIS group than in the non-EIS group (26.9% vs 73.5%). Thirty-one of the EIS and 44 of the non-EIS treatment patients, mainly Child's B and C patients, died within 2 months after the first bleeding. In the short term, EIS decreased the mortality due to esophageal variceal bleeding, but the survivors still had to face hepatic failure and tumor growth. Thus, benefits of EIS were noted on short- but not on long-term survival. The mean survival times were 2.38 months for the EIS group and 1.79 months for the non-EIS group. Since EIS had no beneficial effects on long-term survival it is doubtful whether sclerotherapy applied to esophageal variceal bleeding in patients with advanced HCC would be worthwhile, as the endoscopic procedure would only add to their suffering.