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Helicase-like transcription factor (HLTF) has been found to be involved in the maintenance of genome stability and tumour suppression, but whether its downregulation in cancers is associated with posttranslational regulation remains unclear. Here, we observed that HLTF was significantly downregulated in hepatocellular carcinoma (HCC) tissues and positively associated with the survival of HCC patients. Mechanistically, the decreased expression of HLTF in HCC was attributed to elevated β-TrCP-mediated ubiquitination and degradation. Knockdown of HLTF enhanced p62 transcriptional activity and mammalian target of rapamycin (mTOR) activation, leading to HCC tumorigenesis. Inhibition of mTOR effectively blocked β-TrCP overexpression- or HLTF knockdown-mediated HCC tumorigenesis and metastasis. Furthermore, in clinical tissues, decreased HLTF expression was positively correlated with elevated expression of β-TrCP, p62, or p-mTOR in HCC patients. Overall, our data not only uncover new roles of HLTF in HCC cell proliferation and metastasis but also reveal a novel posttranslational modification of HLTF by β-TrCP, indicating that the β-TrCP/HLTF/p62/mTOR axis may be a new oncogenic driver involved in HCC development. This finding provides a potential therapeutic strategy for HCC patients by targeting the β-TrCP/HLTF/p62/mTOR axis.

The hepatic artery is the only blood source nourishing the biliary duct and associated with biliary complication after liver transplantation (LT). Gastroduodenal artery (GDA) disconnection increased proper hepatic artery flow. Whether this procedure attenuates biliary non-anastomotic stricture (NAS) is not clear.A total of 241 patients with LT were retrospectively analyzed. The patients were divided into the GDA disconnection (GDA-) and GDA preservation (GDA+) groups. Propensity score matching (PSM) was administrated to reduce bias. Logistic regression was conducted to analyze risk factors for biliary NAS before and after PSM. Postoperative complications were compared. Kaplan-Meier survival analysis and log-rank tests were performed to compare overall survival.In all, 99 patients (41.1%) underwent GDA disconnection, and 49 (20.3%) developed NAS. Multivariable logistic regression revealed that GDA preservation (OR = 2.24, 95% CI: 1.11-4.53; P = 0.025) and model for end-stage liver disease (MELD) score15 (OR = 2.14, 95% CI: 1.12-4.11; P = 0.022) were risk factors for biliary NAS. PSM provided 66 pairs using 1:2 matching method, including 66 GDA disconnection and 99 GDA preservation patients. Multivariable logistic regression after PSM also showed that GDA preservation (OR = 3.15, 95% CI: 1.26-7.89; P = 0.014) and MELD score15 (OR = 2.41, 95% CI: 1.08-5.36; P = 0.031) were risk factors for NAS. When comparing complications between the two groups, GDA preservation was associated with a higher incidence of biliary NAS before and after PSM (P = 0.031 and 0.017, respectively). In contrast, other complications including early allograft dysfunction (P = 0.620), small-for-size graft syndrome (P = 0.441), abdominal hemorrhage (P = 1.000), major complications (Clavien-Dindo grade ≥ 3, P = 0.318), and overall survival (P = 0.088) were not significantly different between the two groups.GDA disconnection during LT ameliorates biliary NAS incidence and may be recommended for application in clinical practice.

Hepatocellular carcinoma (HCC) is reported to associate with abnormal expression of SCF E3 ubiquitin ligases. FBXW10, an F-box protein of the E3 ubiquitin ligases, was abnormally regulated in HCC patients. However, whether FBXW10 is associated with HCC has not yet been evaluated. Here, we analyzed the associations between overall survival and various risk factors in 191 HCC tissues. Univariate and multivariate analyses demonstrated that FBXW10 was an independent risk factor related to HCC prognosis. The results showed that FBXW10, gender and tumor state were strongly associated with overall survival in HCC patients. Furthermore, high expression of FBXW10 was associated with poor survival among male HCC patients but not female HCC patients. FBXW10 was more highly expressed in male HCC tissues and more strongly related to vascular invasion in male HCC patients. Consistent with these findings, the male FBXW10-Tg(+) mice were more susceptible to tumorigenesis, changes in regenerative capacity, and liver injury and inflammation but not changes in liver function than FBXW10-Tg(–) mice. FBXW10 promoted cell proliferation and migration in HCC cell lines. Our findings reveal that FBXW10, an independent risk factor for HCC, promotes hepatocarcinogenesis in male patients, and is also a potential prognostic marker in male patients with HCC.

Background Iatrogenic biliary injury (IBI) following laparoscopic cholecystectomy (LC) is the most serious iatrogenic complications. Little is known whether LC-IBI would lead to surgeon's severe mental distress (SMD). Methods A cross-sectional survey in the form of electronic questionnaire was conducted among Chinese general surgeons who have caused LC-IBI. The six collected clinical features relating to mental distress included: 1) feeling burnout, anxiety, or depression, 2) avoiding performing LC, 3) having physical reactions when recalling the incidence, 4) having the urge to quit surgery, 5) taking psychiatric medications, and 6) seeking professional psychological counseling. Univariable and multivariable analyses were performed to identify risk factors of SMD, which was defined as meeting ≥3 of the above-mentioned clinical features. Results Among 1466 surveyed surgeons, 1236 (84.3%) experienced mental distress following LC-IBI, and nearly half (49.7%, 614/1236) had SMD. Multivariable analyses demonstrated that surgeons from non-university affiliated hospitals (OR:1.873), patients who required multiple repair operations (OR:4.075), patients who required hepaticojejunostomy/partial hepatectomy (OR:1.859), existing lawsuit litigation (OR:10.491), existing violent doctor–patient conflicts (OR:4.995), needing surgeons' personal compensation (OR:2.531), and additional administrative punishment by hospitals (OR:2.324) were independent risk factors of surgeon's SMD. Conclusion Four out of five surgeons experienced mental distress following LC-IBI, and nearly half had SMD. Several independent risk factors of SMD were identified, which could help to make strategies to improve surgeons' mental well-being.

NUF2 (NUF2, Ndc80 kinetochore complex component) plays an important role in kinetochore-microtubule attachment. It has been reported that NUF2 is associated with multiple human cancers. However, the functional role of NUF2 in pancreatic cancer remains unclear. In this study, we found that NUF2 expression was stronger in tumour tissues than in normal pancreatic tissues, and its overexpression could be related to poor prognosis. Moreover, NUF2 was highly expressed in several human pancreatic cancer cell lines. We took advantage of lentivirus-mediated siRNA (small interfering RNA) to suppress NUF2 expression in PANC-1 and Sw1990 cell lines aiming to investigate the role of NUF2 in pancreatic cancer. NUF2 silencing by RANi (RNA interference) reduced the proliferation and colony formation ability of pancreatic cancer cells in vitro. Cell cycle analysis showed that NUF2 knockdown induced cell cycle arrest at G0/G1 phase via suppression of Cyclin B1, Cdc2 and Cdc25A. More importantly, NUF2 silencing was able to alleviate in vivo tumourigenesis in pancreatic cancer xenograft nude mice. Collectively, the present study indicates that the siRNA-mediated knockdown against NUF2 may be a promising therapeutic method for the treatment of pancreatic cancer., NUF2 is overexpressed in pancreatic cancer tissues and cell lines. siRNA-mediated knockdown against NUF2 resulted in a significant reduction in pancreatic cancer cell growth both in vitro and in vivo. NUF2 may be a promising therapeutic target in pancreatic cancer.

The relationships between epithelial-to-mesenchymal transition (EMT), hepatitis B virus X protein (HBx), and the non-receptor tyrosine kinase c-Src were investigated. The HBx gene transfected SMMC-7721 cells underwent morphological changes from a classic epithelial morphology to a spindle-like shape. The HBx transfection increased the invasive potential of these cells. When the transfected cells were exposed to the c-Src kinase inhibitor PP2, the cells recovered their original epithelial morphology and the mRNA and protein expression of epithelial and mesenchymal markers returned to the parental cell levels. Our data suggest that activated c-Src played a critical role in the HBx-induced EMT of SMMC-7721 cells.

To evaluate the mechanism of increased invasion and migration of hepatocellular carcinoma (HCC) cells induced by vascular endothelial growth factor receptor-1 (VEGFR-1) activation.Vascular endothelial growth factor-B (VEGF-B) was used to induce and stimulate hepatocellular carcinoma cell line MHCC97-H. Morphologic changes of MHCC97-H were investigated. The expression of E-cadherin and alpha-catenin (two epithelial markers) and Vimentin and N-cadherin (two mesenchymal markers) was detected by reverse transcriptase polymerase chain reaction (RT- PCR), western blotting, and immunofluorescence staining. Cell invasion and migration test was performed.Treatment of MHCC97-H cells with VEGF-B led to morphologic changes characteristic of epithelial to mesenchymal transition (EMT), including loss of polarity, increased intercellular separation, and the presence of pseudopodia. Expression of the epithelial adhesion molecules, including E-cadherin and alpha-catenin, was decreased after VEGF-B treatment. Conversely, an increase in the expression of the mesenchymal cell markers, including N-cadherin and vimentin, was observed after VEGF-B treatment (P less than 0.05). VEGF-B-treated cells exhibited a change in E-cadherin from an organized, membrane-bound structure to a disorganized state in which it was noted to be dispersed throughout the cytoplasm. Pretreatment with VEGFR-1 blocking antibody 18F1 inhibited the change in localization of E-cadherin induced by VEGF-B treatment. The ability of invasion and migration of MHCC97-H was enhanced by VEGF-B reatment (P less alpha 0.05).Increased invasion and migration of HCC cells induced by VEGFR-1 activation was mediated by epithelial to mesenchymal transition.

To examine whether or not vascular endothelial growth factor (VEGF) and its receptors were expressed in hepatocellular carcinoma cell lines with various metastatic potentialities.Reverse transcription-polymerase chain reaction (RT-PCR), enzyme linked immunosorbent assay (ELISA) and Western blot were employed to study the expressions of VEGFR-1, VEGFR-2, VEGF-A and VEGF-B in four hepatocellular carcinoma cell lines MHCC97-H, MHCC97-L, SMMC7721 and HepG-2 and one normal liver cell line L-02.Three hepatocellular carcinoma cell lines (MHCC97-H, MHCC97-L, SMMC7721) expressed VEGFR-1 mRNA and their proteins. The expression level of VEGFR-1 in MHCC97-H was higher than that in MHCC97-L (P less than 0.05) and the expression level of VEGFR-1 in MHCC97-L was higher than that in SMMC7721 (P less than 0.05). No expression of VEGFR-1 was found in HepG-2 or L-02. All four hepatocellular carcinoma cell lines and the L-02 cell line expressed VEGFR-2 mRNA and the protein, as well as the VEGFR-1 ligands VEGF-A and VEGF-B. The expression level of VEGFR-2 in all tested hepatocellular carcinoma cell lines and normal liver cell line L-02 showed no significant differences (P more than 0.05).VEGFR-1 was expressed in 4 hepatocellular carcinoma cell lines with various metastatic potentialities. The expression levels appeared to be positively correlated with the potentialities of metastasis of the hepatocellular carcinoma cell lines. VEGFR-1 may relate to the invasiveness and metastatic potential of the hepatocellular carcinoma.

To discuss the influence of cold preservation on intrahepatic biliary microcirculation.Male Sprague Dawley (SD) rats were divided into 3 groups:cold preserve 1 h group (CP 1 h group), cold preserve 24 h group (CP 24 h group) and sham operation group (SO group). Five time points were determined as 0 h, 1 h, 6 h, 24 h and 72 h postoperation. Morphology was observed under light microscope. Dark microspheres were injected into hepatic artery and the number of microspheres in liver portal areas was measured under light microscope. The expressions of eNOS, ET-1 and ICAM-1 in microvascular endothelial cells of hepatic portal area were measured by immunofluorescence double staining technique and in situ hybridization histochemistry.The histological changes of intrahepatic bile duct were more severe in CP 24 h group than in CP 1 h group. The number of microspheres in implanted liver portal areas was increased significantly in CP 24 h group than in CP 1 h group at the same time point. Compared with CP 1 h group, the expression of eNOS in CP 24 h group significantly reduced after liver transplantation, while the expressions of ET-1 and ICAM-1 in CP 24 h group were significantly increased after liver transplantation. The changes of their mRNA expressions were the approximately same as well as their proteins expressions.Cold preservation brings obvious changes of intrahepatic biliary microcirculation and function of vascular endothelial cell after liver transplantation. The obstruction of microcirculation might play an important role in the reperfusion injury after cold preserve of intrahepatic biliary during liver transplantation.

To investigate the cause, prophylaxis, and management of ischemic-type biliary lesion (ITBL) after orthotopic liver transplantation.The clinical data of 212 operations of orthotopic liver transplantation on 209 patients, 184 males and 25 females, aged 44.5 (18-69), including 3 patients undergoing secondary liver transplantation, between February 1999 and August 2004 were reviewed retrospectively.ITBL occurred in 14 patients (6.6%), among whom 5 had biliary lesions of hepatic bifurcation, 3 had intrahepatic biliary lesions, and 6 had multiple extrahepatic and intrahepatic biliary lesions. The incidence rate of ITBL among the recipients of liver in cold storage for more than 10 hours, with donor-recipient ABO blood type incompatibility, with postoperative hepatic arterial lesions, and with hepatitis B related hepatic failure as the primary disease were 9.8% (10/102), 22.2% (2/9), 40% (2/5), and 14.6% (7/48) respectively. The 14 patients with ITBL were managed with conservative treatment, endoscopy, Roux-en-Y anastomosis, or re-transplantation. Seven of the 14 patients were cured, the condition of 5 patients was improved, and 1 patient died with a mortality of 7.1% (1/14). The incidence of ITBL-related graft loss was 23% (3/14).It is crucial to avoid too long preservation time of donor liver and donor-recipient ABO blood type incompatibility, and timely management of postoperative hepatic arterial lesions so as to prevent ITBL. Proper treatment for ITBL should be chosen according to the specific conditions of the lesion of biliary duct tree.

To evaluate the role of orthotopic liver transplantation (OLT) in treating hepatocellular carcinoma (HCC).Data of 92 consecutive orthotopic liver transplantations (OLTs) performed during January 1999 and February 2005 at our institution were analyzed.Of the 92 recipients, 8 HCC patients were stage I, 13 were stage II, 12 stage III and 59 stage IV (UICC TNM staging system). Overall 1-, 2-, 3-, 5-year patient survival rates were 65.3%, 27.0%, 20.0%, and 6.9%, respectively. When OLT indications were considered, best recipients survival was obtained in stage I patients (100.0%, 100.0%, 66.7%, and 50.0% at 1, 2, 3, and 5 years, respectively) and stage II patients (85.7%, 66.7%, and 66.7% at 1, 2 and 3 years, respectively). Whereas, 1, 2, 3 and 5-year recipients survival rates were 50.0%, 0, 0, 0 in stage III patients, and 58.1%, 20.0%, 13.0% and 5.0% in stage IV patients.The prognosis of different stages of HCC patients who underwent OLT was significantly different. The OLT recipients with HCC should be strictly selected. Long-term recipient survival could be obtained in stage I and stage II patients.

To evaluate the effects of primary duct closure and T-tube drainage in laparoscopy choledochotomy to treat the common bile duct stones.The enrollment of the patients was in accordance with 6 criteria. 55 patients with cholecystolithiasis and secondary common bile duct stones from January 2000 to February 2003 were treated with laparoscopic choledochotomy. The patients were randomly divided into two groups: primary duct closure group and T-tube drainage group. Their all data were recorded and studied prospectively,and patients were followed up after discharge.There were 27 patients and 28 patients in primary duct closure group and T-tube drainage group respectively. The operation time and the results of following up between the two groups had no significant difference. Compared with T-tube drainage group, primary duct closure group had less the total quantity of postoperative transfusion and hospital costs, shorter postoperative hospital stay. The incidence of postoperative complications in primary duct closure group was 11.1% (3/27), and all of them were biliary complications. The incidence of postoperative complications in T-tube drainage group was 28.6% (8/28), and seven of them were biliary complications. The incidence of severe complications that needed reoperations was 10.7% (93/28), and all of them were caused by T-tubes. There was no mortality in two groups.The primary duct closure in laparoscopic choledochotomy can avoid the deficiency of T-tube drainage, and it is feasible and safe and lower complications in treating the common bile duct stones, so we advocate it in appropriate cases.

To investigate whether neuron-glial antigen 2 (NG2) could be an effective prognostic marker in hepatocellular carcinoma (HCC).NG2 expression was semi-quantitatively scored from the immunohistochemistry (IHC) data based on the number of positive cells and the staining intensity. A total of 132 HCC specimens and 96 adjacent noncancerous tissue samples were analyzed by IHC for NG2 protein expression. To confirm the NG2 expression levels observed by IHC, we measured NG2 expression in 30 randomly selected tumor and adjacent noncancerous tissue samples by quantitative real-time polymerase chain reaction and Western blot. The correlations between NG2 protein expression and the clinicopathological features of HCC patients were analyzed using the χ (2) test. To assess the prognostic value of NG2 for HCC, the association between NG2 expression and survival was analyzed using the Kaplan-Meier method with the log-rank test. To further evaluate the prognostic value of NG2 expression, a Cox multivariate proportional hazards regression analysis was performed with all the variables to derive risk estimates related to disease-free and overall survival and to control for confounders.High NG2 expression was observed in significantly more primary tumor samples (63.6%; 84/132) compared with the adjacent noncancerous tissue samples (28.1%; 27/96) (P0.0001). Moreover, high NG2 protein expression was closely associated with tumor differentiation (χ (2) = 9.436, P = 0.0089), recurrence (χ (2) = 5.769, P = 0.0163), tumor-node-metastasis (TNM) stage (χ (2) = 8.976, P = 0.0027), and invasion (χ (2) = 5.476, P = 0.0193). However, no significant relationship was observed between NG2 protein expression in HCC and other parameters, such as age, sex, tumor size, serum alpha fetoprotein (AFP), tumor number, or tumor capsule. The log-rank test indicated a significant difference in the overall survival of HCC patients with high NG2 expression compared with those with low NG2 expression (29.2% vs 9.5%, P0.001). Moreover, NG2 expression in HCC tissue significantly correlated with disease-free survival (15.2% vs 6.7%, P0.001). Multivariate analysis showed that NG2 expression (HR = 2.035, P = 0.002), serum AFP (HR = 1.903, P = 0.003), TNM stage (HR = 2.039, P = 0.001), and portal vein invasion (HR = 1.938, P = 0.002) were independent prognostic indicators for OS in HCC patients. Furthermore, NG2 expression (HR = 1.974, P = 0.003), serum AFP (HR = 1.767, P = 0.008), TNM stage (HR = 2.078, P = 0.001), tumor capsule (HR = 0.652, P = 0.045), and portal vein invasion (HR = 1.941, P = 0.002) were independent prognostic indicators for DFS in HCC patients.The up-regulation of NG2 is associated with poor prognosis in HCC. Therefore, NG2 could be useful as an additional prognostic marker to increase the resolution of traditional approaches.