Weifeng Tan

Sorafenib extends the survival time of patients with multiple recurrences of hepatocellular carcinoma after liver transplantation

Aim:To determine the efficacy and toxicities of sorafenib in the treatment of patients with multiple recurrences of hepatocellular carcinoma (HCC) after liver transplantation in a Chinese population.Methods:Twenty patients with multiple recurrences of HCC after liver transplantation were retrospectively studied. They received either transarterial chemoembolization (TACE) or TACE combined with sorafenib.Results:The median survival times (MST) after multiple recurrences was 14 months (TACE+sorafenib group) and 6 months (TACE only group). The difference was significant in MST between the two groups (P=0.005). The TACE + sorafenib group had more stable disease (SD) patients than the TACE group. The most frequent adverse events of sorafenib were hand–foot skin reaction and diarrhea. In the univariate analysis, preoperative bilirubin and CHILD grade are found to be significantly associated with tumor-free survival time, the survival time after multiple recurrences and overall survival time. TACE+sorafenib group showed a better outcome than single TACE treatment group. In the multivariate COX regression modeling, the preoperative high CHILD grade was found to be a risk factor of tumor-free survival time. In addition, the preoperative high bilirubin grade was also found to be a risk factor of survival time after recurrence and overall survival time. Furthermore, survival time after recurrence and overall survival time were also associated with therapeutic schedule, which was indicated by the GROUP.Conclusion:Treatment with TACE and sorafenib is worthy of further study and may have more extensive application prospects.

hSulf-1 Gene Exhibits Anticancer Efficacy through Negatively Regulating VEGFR-2 Signaling in Human Cancers

Background Human sulfatase 1 (hSulf-1) is a heparin-degrading endosulfatase that desulfates cell surface heparan sulfate proteoglycans (HSPGs) in extracellular matrix and negatively modulates heparin-binding growth factor and cytokine signaling in cell proliferation. But hSulf-1 function is more complicated, and its molecular mechanism has not been well known. Principal Findings To further investigate the functions of hSulf-1 gene in regulating the vascular endothelial growth factor receptor (VEGFR) signaling, a series of vectors expressing hSulf-1, hSulf-1 small hairpin RNA (shRNA) and VEGFR-2 shRNA were generated. hSulf-1 re-expression could downregualte the VEGFR-2 phosphorylation and inhibit cancer cell proliferation both in ovarian and hepatocellular cancer cell lines. Knockdown of hSulf-1 expression by hSulf-1 shRNA enhanced the recovery of high levels of phosphorylated VEGFR-2, and knockdown of VEGFR-2 expression by VEGFR-2 shRNA inhibited the proliferation activity of cancer cells in vitro to some extent. In human cancer xenografts in nude mice, tumor growth was inhibited markedly after injections of adenovirus expressing hSulf-1, with the tumor inhibition rates of 46.19% and 49.56% in ovarian and hepatocellular tumor models, respectively. hSulf-1 expression significantly reduced tumor microvessel density. Conclusions The results demonstrated that hSulf-1 re-expression both in ovarian and hepatocellular cancer cells induces antitumor efficacy by attenuating the phosphorylation of VEGFR-2 and suppressing angiogenesis. Therefore, hSulf-1-mediated antiproliferation and antiangiogenesis could be a reasonable approach for cancer therapy.

Inhibitory effect of Survivin promoter-regulated oncolytic adenovirus carrying P53 gene against gallbladder cancer

Gene therapy has become an important strategy for treatment of malignancies, but problems remains concerning the low gene transferring efficiency, poor transgene expression and limited targeting specific tumors, which have greatly hampered the clinical application of tumor gene therapy. Gallbladder cancer is characterized by rapid progress, poor prognosis, and aberrantly high expression of Survivin. In the present study, we used a human tumor‐specific Survivin promoter‐regulated oncolytic adenovirus vector carrying P53 gene, whose anti‐cancer effect has been widely confirmed, to construct a wide spectrum, specific, safe, effective gene‐viral therapy system, AdSurp‐P53. Examining expression of enhanced green fluorecent protein (EGFP), E1A and the target gene P53 in the oncolytic adenovirus system validated that Survivin promoter‐regulated oncolytic adenovirus had high proliferation activity and high P53 expression in Survivin‐positive gallbladder cancer cells. Our in vitro cytotoxicity experiment demonstrated that AdSurp‐P53 possessed a stronger cytotoxic effect against gallbladder cancer cells and hepatic cancer cells. The survival rate of EH‐GB1 cells was lower than 40% after infection of AdSurp‐P53 at multiplicity of infection (MOI) = 1 pfu/cell, while the rate was higher than 90% after infection of Ad‐P53 at the same MOI, demonstrating that AdSurp‐P53 has a potent cytotoxicity against EH‐GB1 cells. The tumor growth was greatly inhibited in nude mice bearing EH‐GB1 xenografts when the total dose of AdSurp‐P53 was 1 × 109 pfu, and terminal dUTP nick end‐labeling (TUNEL) revealed that the apoptotic rate of cancer cells was (33.4 ± 8.4)%. This oncolytic adenovirus system overcomes the long‐standing shortcomings of gene therapy: poor transgene expression and targeting of only specific tumors, with its therapeutic effect better than the traditional Ad‐P53 therapy regimen already on market; our system might be used for patients with advanced gallbladder cancer and other cancers, who are not sensitive to chemotherapy, radiotherapy, or who lost their chance for surgical treatment.

Risk factors of intra-abdominal bacterial infection after liver transplantation in patients with hepatocellular carcinoma

OBJECTIVE To explore the risk factors of intra-abdominal bacterial infection (IAI) after liver transplantation (LT) in patients with hepatocellular carcinoma (HCC). METHODS A series of 82 HCC patients who received LT surgeries in our department between March 2004 and April 2010 was recruited in this study. Then we collected and analyzed the clinical data retrospectively. Statistical analysis system (SPSS) software was adopted to perform statistical analysis. Chi-square test, t-test and Wilcoxon rank sum test were used to analyze the clinical data and compute the significance of the incidences of early-stage IAI after LT for HCC patients. Binary logistic regression was performed to screen out the risk factors, and multiple logistic regression analyses were performed to compute the independent risk factors. RESULTS A series of 13 patients (13/82, 15.9%) had postoperative IAI. The independent risk factors of postoperative intra-abdominal bacterial infections after LT for HCC patients were preoperative anemia [Hemoglobin (HGB) <90 g/L] and postoperative abdominal hemorrhage (72 hours >400 mL), with the odds ratios at 8.121 (95% CI, 1.417 to 46.550, P=0.019) and 5.911 (95% CI, 1.112 to 31.432, P=0.037). CONCLUSIONS Postoperative IAI after LT in patients with HCC was a common complication. Preoperative moderate to severe anemia, as well as postoperative intra-abdominal hemorrhage more than 400 mL within the first 72 hours might independently indicate high risk of IAI for these patients.

Early control of short hepatic portal veins in isolated or combined hepatic caudate lobectomy

BACKGROUND Caudate lobectomy has long been considered technically difficult. This study aimed to elaborate the significance of early control of short hepatic portal veins (SHPVs) in isolated hepatic caudate lobectomy or in hepatic caudate lobectomy combined with major partial hepatectomy, and to describe the anatomical characteristics of SHPVs. METHODS The data of 117 patients who underwent either isolated or combined caudate lobectomy by the same team of surgeons from 2005 to 2009 were retrospectively analyzed. From 2005 to 2007 (group A, n=55), we carried out early control of short hepatic veins (SHVs) only; from 2008 to 2009 (group B, n=62), we carried out early control of both SHVs and SHPVs. The two groups were compared to evaluate which surgical procedure was better. A detailed anatomical study was then carried out on the last 25 consecutive patients in group B to study the number and distribution of SHPVs during surgery. RESULTS Patients in group B had less intra-operative blood loss, less impairment of liver function, shorter postoperative hospital stay, fewer postoperative complications and required less blood transfusion (P<0.05). The number of SHPVs in the 25 patients was 183, with 7.3+/-2.7 per patient. The diameters of SHPVs were 1 to 4 mm. On average, 3.4 SHPVs/patient came from the left portal vein, 2.2 from the bifurcation, 1.4 from the right portal vein, and 0.3 from the main portal vein. On average, 3.3 SHPVs/patient supplied segment I of the liver, 0.4 for segment II, 2.1 for segment IV, 1.4 for segment V and 0.1 for segment VI. CONCLUSION Early control of SHPVs in isolated or combined hepatic caudate lobectomy may be a useful method to decrease surgical risk and improve postoperative recovery.

Efficacy of infusing cold fluid through the bile duct in cooling the liver

Objective To evaluate the liver cooling effect of infusing cold fluid through the bile duct in liver injury models. Methods A total of 14 BA-MA mini-pigs were randomized into two groups: control group( n = 7) and hypothermia group( n = 7). All the animals underwent tracheal intubation under general anaesthesia and liver injury was created after opening the abdomen. The liver bleeding was allowed for 15 min,and then initial treatment was given by packing the liver injury and infusing fluid for 1 hour before suturing the wound. Then the cold fluid( 4℃ lactated Ringers solution) was infused through the bile duct in the hypothermia group, with no further treatment given to the control group. The body and local liver temperature and hemodynamic variations were recorded. Blood samples were collected from the front vena cava to perform liver function tests. The liver specimens of animals were subjected to histopathological evaluation at the end of the experiment. Results The primary bleeding volume,heart rate and mean artery pressure at different time points were not significantly different between the two groups. After 2 hours intervention,the liver temperature dropped by( 1. 36 ±0. 80) ℃ in the control group and by( 2. 72 ±1. 01) ℃ in the hypothermia group,with significant difference found between the two groups( P = 0. 020); at the same time point,the body temperature dropped by( 0. 89 ± 0. 76) ℃ and( 0. 97 ± 0. 97) ℃, respectively; showing no significant difference between the two groups( P = 0. 87). Liver function examination showed that at the end of infusion aspartate transaminase( AST) in the hypothermia group was significantly lower than that in the control group( P 0. 05). The inflammatory cell infiltration of liver tissues in the hypothermia group was slighter than that in the control group. Conclusion Infusion with cold 4℃ lactated Ringers solution has a protective effect on liver trauma by inducing liver hypothermia to promote the recovery of liver function and alleviate liver inflammation reaction.

Lentiviral vector-mediated RNA interference of HBs gene inhibits replication of HBV: Lentiviral vector-mediated RNA interference of HBs gene inhibits replication of HBV

Objective:To construct a lentiviral vector(LV) of RNA interference(RNAi) targeting HBs gene and to observe its effect on the replication of HBV and expression of antigens.Methods: The effective sequence of siRNA targeting HBs gene was confirmed in our previous study.The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the lentivirus vector(pGCLM-GFP).The resulting lentivirus vector containing HBs shRNA was named LVshHBs,and it was confirmed by PCR and sequencing.293T cells were cotransfected with lentivirus vector pGCLM-GFP,pHelper 1.0 and pHelper 2.0.All virus stocks were produced by calcium phosphate-mediated transfection.The titer of virus was tested according to the expression level of GFP.HepG2.2.15 cells were infected with LVshHBs and the supernatant of the cells was subjected to ELISA,Western blotting analysis and HBV DNA quantitative analysis.Results: PCR and DNA sequencing demonstrated that the LVshHBs-producing HBs shRNA was successfully constructed.The titer of concentrated virus was 5×108-2×109 TU /ml.The inhibitory effect was efficient and the corresponding viral transcript and expression of antigens were decreased after infection.The inhibitory effect was observed 4 days after infection and peaked 9 days after the initial treatment with RNAi.Secreted HBsAg was reduced by 70% in cell culture compared with the negative control,which is also confirmed by Western blotting and real-time PCR.After quantification of HBV DNA,the level of DNA relative to the controls was also significantly reduced after RNAi treatment(P0.05).Conclusion: The lentivirus RNAi vector of HBs has been successfully constructed.The lentiviral microRNA-based RNAi targeting HBs can specifically mediate the down-regulation of HBs expression,inhibiting HBV replication and antigen expression.