Hypomethylation of glycine dehydrogenase promoter in peripheral blood mononuclear cells is a new diagnostic marker of hepatitis B virus-associated hepatocellular carcinoma

Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.

Multifaceted roles of lymphatic and blood endothelial cells in the tumor microenvironment of hepatocellular carcinoma:A comprehensive review

The tumor microenvironment is a complex network of cells,extracellular matrix,and signaling molecules that plays a critical role in tumor progression and metastasis.Lymphatic and blood vessels are major routes for solid tumor metastasis and essential parts of tumor drainage conduits.However,recent studies have shown that lymphatic endothelial cells(LECs)and blood endothelial cells(BECs)also play multifaceted roles in the tumor microenvironment beyond their structural functions,particularly in hepatocellular carcinoma(HCC).This comprehensive review summarizes the diverse roles played by LECs and BECs in HCC,including their involvement in angiogenesis,immune modulation,lymphangiogenesis,and metastasis.By providing a detailed account of the complex interplay between LECs,BECs,and tumor cells,this review aims to shed light on future research directions regarding the immune regulatory function of LECs and potential therapeutic targets for HCC.

Treatment of fat-poor renal angiomyolipoma with ectopic blood supply by fluorescent laparoscopy:A case report and review of literature

BACKGROUND Renal angiomyolipoma and renal cell carcinoma are the most common benign and malignant tumors of the kidney respectively,and the preoperative differential diagnosis is crucial due to the wide difference in treatment methods.Fat-poor renal angiomyolipoma is a relatively rare type of in renal angiomyolipoma.Its fat imaging features are not obvious,and it is easily misdiagnosed as renal cell carcinoma.CASE SUMMARY We report the case of a 41-year-old man who complained of osphyalgia.Subsequent abdominal computed tomography scans revealed that a heterogeneous mass was seen in the lower pole of the right kidney,with the size of about 53 mm×47 mm.And showed two right renal arteries,with the mass supplied by an ectopic vessel from the abdominal aorta.Fluorescent laparoscopic blockade of the right renal heterotopic artery and partial nephrectomy was performed.Based on histological and immunohistochemical findings,the tumor was diagnosed as fatpoor renal angiomyolipoma.CONCLUSION The use of fluorescent laparoscopy can effectively help intraoperative management,and the fluorescence pattern provided by intravenous indocyanine green can help suggest the final diagnosis,effectively guide the surgical decisionmaking,and avoid preoperative imaging diagnosis leading to nephrectomy for benign renal tumors,through fluorescent navigation of tumor supply vessel precise block,minimize the loss of renal function.

Blood neutrophil-lymphocyte ratio predicts survival after hepatectomy for hepatocellular carcinoma: A propensity score-based analysis

AIM: To investigate whether an elevated preoperative neutrophil-to-lymphocyte ratio(NLR) can predict poor survival in patients with hepatocellular carcinoma(HCC).METHODS: We retrospectively reviewed 526 patients with HCC who underwent surgery between 2004 and 2011.RESULTS: Preoperative NLR ≥ 2.81 was an independent predictor of poor disease-free survival(DFS, P < 0.001) and overall survival(OS, P = 0.044). Compared with patients who showed a preoperative NLR < 2.81 and postoperative increase, patients who showed preoperative NLR ≥ 2.81 and postoperative decrease had worse survival(DFS, P < 0.001; OS, P < 0.001). Among patients with preoperative NLR ≥ 2.81, survival was significantly higher among those showing a postoperative decrease in NLR than among those showing an increase(DFS, P < 0.001; OS, P < 0.001). When elevated, alpha-fetoprotein(AFP) provided no prognostic information, and so preoperative NLR ≥ 2.81 may be a good complementary indicator of poor OS whenever AFP levels are low or high.CONCLUSION: Preoperative NLR ≥ 2.81 may be an indicator of poor DFS and OS in patients with HCC undergoing surgery. Preoperative NLR ≥ 2.81 may be a good complementary indicator of poor OS when elevated AFP levels provide no prognostic information.

Controversy over the use of intraoperative blood salvage autotransfusion during liver transplantation for hepatocellular carcinoma patients

Intraoperative blood salvage autotransfusion (IBSA) is used in various surgical procedures. However, because of the risk of reinfusion of salvaged blood contaminated by tumor cells, the use of IBSA in hepatocellular carcinoma (HCC) patients undergoing liver transplantation (LT) is controversial. The critical points include whether tumor cells can be cleared by IBSA, whether IBSA increases the risk of recurrence or metastasis, and what are the indications for IBSA. Moreover, is it warranted to take the risk of tumor dissemination by using IBSA to avoid allogeneic blood transfusion? Do the remaining tumor cells after additional filtration by leukocyte depletion filters still possess potential tumorigenicity? Does IBSA always work well? We have reviewed the literature and tried to address these questions. The available data indicate that IBSA is safe in LT for HCC, but randomized, controlled and prospective trials are urgently required to clarify the uncertainty.

Hepatic blood inflow occlusion without hemihepatic artery control in treatment of hepatocellular carcinoma

AIM:To investigate the clinical significance of hepatic blood inflow occlusion without hemihepatic artery control (BIOwHAC) in the treatment of hepatocellular carcinoma (HCC).METHODS:Fifty-nine patients with HCC were divided into 3 groups based on the technique used for achieving hepatic vascular occlusion:group 1,vascular occlusion was achieved by the Pringle maneuver (n=20);group 2,by hemihepatic vascular occlusion (HVO) (n=20);and group 3,by BIOwHAC (n=19).We compared the procedures among the three groups in term of operation time,intraoperative bleeding,postoperative liver function,postoperative complications,and length of hospital stay.RESULTS:There were no statistically significant differences (P > 0.05) in age,sex,pathological diagnosis,preoperative Child's disease grade,hepatic function,and tumor size among the three groups.No intraoperative complications or deaths occurrred,and there were no significant intergroup differences (P > 0.05) in intraoperative bleeding,hepatic function change 3 and 7 d after operation,the incidence of complications,and length of hospital stay.BIOwHAC and Pringle maneuver required a significantly shorter operation time than HVO;the difference in the serum alanine aminotransferase or aspartate aminotransferase levels before and 1 d after operation was more significant in the BIOwHAC and HVO groups than in the Pringle maneuver group (P < 0.05).CONCLUSION:BIOwHAC is convenient and safe;this technique causes slight hepatic ischemia-reperfusion injury similar to HVO.

Multi-parameter gene expression profiling of peripheral blood for early detection of hepatocellular carcinoma

AIM In our previous study, we have built a nine-gene(GPC3, HGF, ANXA1, FOS, SPAG9, HSPA1 B, CXCR4, PFN1, and CALR) expression detection system based on the Ge XP system. Based on peripheral blood and Ge XP, we aimed to analyze the results of genes expression by different multi-parameter analysis methods and build a diagnostic model to classify hepatocellular carcinoma(HCC) patients and healthy people.METHODS Logistic regression analysis, discriminant analysis, classification tree analysis, and artificial neural network were used for the multi-parameter gene expression analysis method. One hundred and three patients with early HCC and 54 age-matched healthy normal controls were used to build a diagnostic model. Fiftytwo patients with early HCC and 34 healthy people were used for validation. The area under the curve, sensitivity, and specificity were used as diagnostic indicators.RESULTS Artificial neural network of the total nine genes had the best diagnostic value, and the AUC, sensitivity, and specificity were 0.943, 98%, and 85%, respectively. At last, 52 HCC patients and 34 healthy normal controls were used for validation. The sensitivity and specificity were 96% and 86%, respectively.CONCLUSION Multi-parameter analysis methods may increase the diagnostic value compared to single factor analysis and they may be a trend of the clinical diagnosis in the future.

Advantage of autologous blood transfusion in surgery for hepatocellular carcinoma

AIM: To evaluate the significance of autologous blood transfusion (AT) in reducing homologous blood transfusion (HT) in surgery for hepatocellular carcinoma (HCC). METHODS: The proportion of patients who received HT was compared between two groups determined by the time of AT introduction; period A (1991-1994, n = 93) and period B (1995-2000, n = 201). Multivariate logistic regression analysis was performed in order to identify independent significant predictors of the need for HT. We also investigated the impact of AT and HT on long-term postoperative outcome after curative surgery for HCC. RESULTS: The proportion of patients with HT was significantly lower in period B than period A (18.9% vs 60.2%, P < 0.0001). Multivariate logistic regression analysis identified AT administration as a significant independent predictor of the need for HT (P < 0.0001). Disease-free survival in patients with AT was comparable to that without any transfusion. Multivariate analysis identified HT administration as an independent significant factor for poorer disease-free survival (P = 0.0380). CONCLUSION: AT administration significantly decreased the need for HT. Considering the postoperative survival disadvantage of HT, AT administration could improve the long-term outcome of HCC patients.

Dynamic alteration of telomerase expression and its diagnostic significance in liver or peripheral blood for hepatocellular carcinoma

AIM： To investigate the dynamic alteration of telomerase expression during development of hepatocellular carcinoma （HCC） and its diagnostic implications in liver tissues or peripheral blood mononuclear cells for HCC. METHODS： Dynamic expressions of liver telomerase during malignant transformation of hepatocytes were observed in Sprague-Dawly （SD） rats fed with 0.05% of 2-fluoenyacetamide （2-FAA）. Total RNA and telomerase were extracted from rat or human liver tissues. The telomerase activities in livers and in circulating blood were detected by a telomeric repeat amplification protocol-enzyme-linked immunosorbent assay （TRAP- ELISA）, and its diagnostic value was investigated in patients with benign or malignant liver diseases. RESULTS： The hepatoma model displayed the dynamic expression of hepatic telomerase during HCC development. The telomerase activities were consistent with liver total RNA levels （r = 0.83, P 〈 0.01） at the stages of degeneration, precancerosis, and cancerization of hepatocytes. In HCC patients, the telomerase levels in HCC tissues were significantly higher than in their adjacent non-cancerous tissues, but liver total RNA levels were lower in the former than in the latter. Although the circulating telomerase of HCC patients was abnormally expressed among patients with chronic liver diseases, the telomerase activity was a non-specific marker for HCC diagnosis, because the incidence was 15.7% in normal control, 25% in chronic hepatitis, 45.9% in liver cirrhosis, and 85.2% in HCC, respectively when absorbance value of telomerase activity was more than 0.2. If the value was over 0.6, the incidence was 60% in HCC group and 0% in any of the others （P 〈 0.01） except in two cases with liver cirrhosis. However, the combination of circulating telomerase with serum alpha-fetoprotein level could increase the positive rate and the accuracy （92.6%, 125 of 135） of HCC diagnosis. CONCLUSION： The overexpression of telomerase is associated with HCC development, and its abnormality in liver tissues or in peripheral blood could be a useful marker for diagnosis and prognosis of HCC.

Blood DNA methylation markers in prospectively identifiedhepatocellular carcinoma cases and controls from Taiwan

AIM: To determine if gene-specific DNA methylation in prospectively collected blood samples is associated with later development of hepatocellular carcinoma(HCC).METHODS: Comparing genome-wide DNA methylation profiles using Illumina Human methylation 450 K arrays, we previously identified a list of loci that were differentially methylated between tumor and adjacent nontumor tissues. To examine if dysregulation of DNAmethylation patterns observed in tumor tissues can be detected in white blood cell(WBC) DNA, we conducted a prospective case-control study nested within a community-based cancer screening cohort in Taiwan with 16 years of follow up. We measured methylation levels in ninety-six loci that were aberrant in DNA methylation in HCC tumor tissues compared to adjacent tissues. Baseline WBC DNA from 159 HCC cases and 312 matched controls were bisulfite treated and assayed by Illumina Bead Array. We used the χ2 test for categorical variables and student's t-test for continuous variables to assess the difference in selected characteristics between cases and controls. To estimate associations with HCC risk, we used conditional logistic regression models stratified on the matching factors to calculate odds ratios(OR) and 95%CI. RESULTS: We found that high methylation level in cg10272601 in WNK2 was associated with increased risk of HCC, with an OR of 1.91(95%CI: 1.27-2.86). High methylation levels in both cg12680131 in TPO and cg22511877 in MYT1 L, however, were associated with decreased risk. The ORs(95%CI) were 0.59(0.39-0.87) and 0.50(0.33-0.77), respectively, for those with methylation levels of cg12680131 and cg22511877 above the median compared with those with levels below the median. These associations were still statistically significant in multivariable conditional logistic regression models after adjusting for hepatitis B virus infection and alcohol consumption. CONCLUSION: These findings support the measurement of methylation markers in WBC DNA as biomarkers of HCC susceptibility but should be replicated in additional prospective studies.

Blood exosomal micro ribonucleic acid profiling reveals the complexity of hepatocellular carcinoma and identifies potential biomarkers for differential diagnosis

BACKGROUND Hepatocellular carcinoma(HCC)is one of the leading causes of cancer-related deaths worldwide,but there is a shortage of effective biomarkers for its diagnosis.AIM To explore blood exosomal micro ribonucleic acids(miRNAs)as potential biomarkers for HCC diagnosis.RESULTS The principal component analysis suggested that daily alcohol consumption could alter the blood exosomal miRNA profiles of hepatitis B virus positive non-HCC patients through miR-3168 and miR-223-3p.The miRNA profiles also revealed the tumor stages of HCC patients.High expression of miR-455-5p and miR-30c-5p,which significantly correlated with better overall survival in tumor tissues,could also be detected in blood exosomes.Two pairs of miRNAs(miR-584-5p/miR-106-3p and miR-628-3p/miR-941)showed a 94.1%sensitivity and 68.4%specificity to differentiate HCC patients from non-HCC patients.The specificity of the combination was substantially influenced by alcohol consumption habits.CONCLUSION This study suggested that blood exosomal miRNAs can be used as new noninvasive diagnostic tools for HCC.However,their accuracy could be affected by tumor stage and alcohol consumption habits.

Frankincense myrrh attenuates hepatocellular carcinoma by regulating tumor blood vessel development through multiple epidermal growth factor receptor-mediated signaling pathways

BACKGROUND In traditional Chinese medicine(TCM),frankincense and myrrh are the main components of the antitumor drug Xihuang Pill.These compounds show anticancer activity in other biological systems.However,whether frankincense and/or myrrh can inhibit the occurrence of hepatocellular carcinoma(HCC)is unknown,and the potential molecular mechanism(s)has not yet been determined.AIM To predict and determine latent anti-HCC therapeutic targets and molecular mechanisms of frankincense and myrrh in vivo.METHODS In the present study,which was based on the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(http://tcmspw.com/tcmsp.php),Universal Protein database(http://www.uniprot.org),GeneCards:The Human Gene Database(http://www.genecards.org/)and Comparative Toxicogenomics Database(http://www.ctdbase.org/),the efficacy of and mechanism by which frankincense and myrrh act as anti-HCC compounds were predicted.The core prediction targets were screened by molecular docking.In vivo,SMMC-7721 human liver cancer cells were transplanted as xenografts into nude mice to establish a subcutaneous tumor model,and two doses of frankincense plus myrrh or one dose of an EGFR inhibitor was administered to these mice continuously for 14 d.The tumors were collected and evaluated:the tumor volume and growth rate were gauged to evaluate tumor growth;hematoxylineosin staining was performed to estimate histopathological changes;immunofluorescence(IF)was performed to detect the expression of CD31,α-SMA and collagen IV;transmission electron microscopy(TEM)was conducted to observe the morphological structure of vascular cells;enzyme-linked immunosorbent assay(ELISA)was performed to measure the levels of secreted HIF-1αand TNF-α;reverse transcription-polymerase chain reaction(RT-qPCR)was performed to measure the mRNA expression of HIF-1α,TNF-α,VEGF and MMP-9;and Western blot(WB)was performed to determine the levels of proteins expressed in the EGFR-mediated PI3K/Akt and MAPK signaling pathways.RESULTS The results of the network pharmacology analysis showed that there were 35 active components in the frankincense and myrrh extracts targeting 151 key targets.The molecular docking analysis showed that both boswellic acid and stigmasterol showed strong affinity for the targets,with the greatest affinity for EGFR.Frankincense and myrrh treatment may play a role in the treatment of HCC by regulating hypoxia responses and vascular system-related pathological processes,such as cytokine-receptor binding,and pathways,such as those involving serine/threonine protein kinase complexes and MAPK,HIF-1 and ErbB signaling cascades.The animal experiment results were verified.First,we found that,through frankincense and/or myrrh treatment,the volume of subcutaneously transplanted HCC tumors was significantly reduced,and the pathological morphology was attenuated.Then,IF and TEM showed that frankincense and/or myrrh treatment reduced CD31 and collagen IV expression,increased the coverage of perivascular cells,tightened the connection between cells,and improved the shape of blood vessels.In addition,ELISA,RT-qPCR and WB analyses showed that frankincense and/or myrrh treatment inhibited the levels of hypoxia-inducible factors,inflammatory factors and angiogenesis-related factors,namely,HIF-1α,TNF-α,VEGF and MMP-9.Furthermore,mechanistic experiments illustrated that the effect of frankincense plus myrrh treatment was similar to that of an EGFR inhibitor with regard to controlling EGFR activation,thereby inhibiting the phosphorylation activity of its downstream targets:the PI3K/Akt and MAPK(ERK,p38 and JNK)pathways.CONCLUSION In summary,frankincense and myrrh treatment targets tumor blood vessels to exert anti-HCC effects via EGFR-activated PI3K/Akt and MAPK signaling pathways,highlighting the potential of this dual TCM compound as an anti-HCC candidate.

Blood oxygen level–dependent functional magnetic resonance imaging can evaluate the efficiency of transcatheter arterial chemoembolization in hepatocellular carcinoma

Hepatocellular carcinoma(HCC) is among the most common malignant tumors worldwide, and transcatheter arterial chemoembolization(TACE) technology has become the first-line treatment for advanced HCC. Another important, recently developed technique is blood oxygen level–dependent functional magnetic resonance imaging(BOLD-fMRI), which utilizes hemoglobin as an endogenous contrast agent and measures deoxygenated hemoglobin content by sampling the oxygen content of tissues, thus reflecting the hemodynamics and pathophysiologic changes in body organs. Currently this technology is being used in patients with liver tumors;that is, it serves as an important tool in follow-up after TACE. The present paper summarizes these developments.

INVESTIGATION OF BLOOD SUPPLY OF BRONCHOGENIC CARCINOMA DERIVING FROM PULMONARY ARTERY

Microfil perfusion technique was used to investigate the blood supply of bronchogenic carcinoma deriving from pulmonary artery on 20 fresh specimens of lung cancer, including 11 squamous carcinoma, 6adenocarcinoma, 1 alveolar cell carcinoma and 2undifferentiated carcinoma cases. The results showed that the appearance and quantity of pulmonary blood supply of bronchogenic carcinoma depended on and changed with the site, activity,growth mode as well as the local condition of tumor nodules; pulmonary artery supplied blood to the Periphery of the tumor and its iunermost part as well; vessels from pulmonary artery in tumor nodules were generally less in number than those in the surrounding normal lung tissues around. The results suggested that the tumor blood supply from pulmonary artery should he evaluated comprehensively and dynamically; during interventional chemotherapy via pulmonary artery, patients should bc selected carefully and the catheter for infusion placed in suitable position so as to gain the best therapeutic effect.

Role of Early Arterial Phase Multislice Helical CT Angiography in Evaluation of Hepatocellular Carcinoma

Objective: To investigate the clinical application of early arterial phase multislice CT angiog raphy (MSCTA) of hepatic vessels in evaluation of middle or advanced stage hepatocellular carcinoma. Methods: Trigger Bolus program was used to carry out MSCTA in early and late arterial phases and portal vein phase with single breath holding. Hepatic vessels were reconstructed from the original images of early arterial phase by post processing. The blood supply of tumor and normal liver tissue and the appearances of venous thrombosis and arteriovenous shunts were analyzed. Results: The MSCTA with early arterial phase could perfectly display the origin, shape and amount of feeding vessels to normal liver tissue and tumor in middle or advanced stage hepatocellular carcinoma. It had the ability of displaying the arteriovenous shunts better than that in conventional dual phased liver scanning. Conclusion: MSCTA of hepatic vessels with early arterial phase acquisition using multislice helical CT in middle or advance stage hepatocellular carcinoma has favorable and promising application. It can be used as an imaging method for comprehensive assessment of the hepatocellular carcinoma before treatment.

Effect of autotransfusion system on tumor recurrence and survival in hepatocellular carcinoma patients

AIM:To investigate the therapeutic efficacy and safety of continuous autotransfusion system(CATS) during liver transplantation of hepatocellular carcinoma patients.METHODS:Eighty-three hepatocellular carcinoma(HCC) patients who underwent liver transplantation with intraoperative CATS(n = 24,CATS group) and without(n = 59,non-CATS group) between April 2006 and November 2011 at the Liver Transplant Institute of Inonu University were analyzed retrospectively.Postoperative HCC recurrence was monitored by measuring alpha-fetoprotein(AFP) levels at 3-mo intervals and performing imaging analysis by thoracoabdominal multidetector computed tomography at 6-month intervals.Inter-group differences in recurrence and correlations between demographic,clinical,and pathological data were assessed by ANOVA and χ 2 tests.Overall and disease-free survivals were calculated by the univariate Kaplan-Meier method.RESULTS:Of the 83 liver transplanted HCC patients,89.2% were male and the overall mean age was 51.3 ± 8.9 years(range:18-69 years).The CATS and nonCATS groups showed no statistically significant differences in age,sex ratio,body mass index,underlying disease,donor type,graft-to-recipient weight ratio,Child-Pugh and Model for End-Stage Liver Disease scores,number of tumors,tumor size,AFP level,Milan and University of California San Francisco selection criteria,tumor differentiation,macrovascular invasion,median hospital stay,recurrence rate,recurrence site,or mortality rate.The mean follow-up time of the nonCATS group was 17.9 ± 12.8 mo,during which systemic metastasis and/or locoregional recurrence developed in 25.4% of the patients.The mean follow-up time for the CATS group was 25.8 ± 15.1 mo,during which systemic metastasis and/or locoregional recurrence was detected in 29.2% of the patients.There was no significant difference between the CATS and non-CATS groups in recurrence rate or site.Additionally,no significant differences existed between the groups in overall or disease-free survival.CONCLUSION:CATS is a safe procedure and may decrease the risk of tumor recurrence in HCC patients.

Efficiency and safety of radiofrequency-assisted hepatectomy for hepatocellular carcinoma with cirrhosis:A single-center retrospective cohort study

AIM: To assess the efficiency and safety of radiofrequencyassisted hepatectomy in patients with hepatocellular carcinoma(HCC) and cirrhosis.METHODS: From January 2010 to December 2013, 179 patients with HCC and cirrhosis were recruited for this retrospective study. Of these, 100 patients who received radiofrequency-assisted hepatectomy(RF+ group) were compared to 79 patients who had hepatectomy without ablation(RF- group). The primary endpoint was intraoperative blood loss. The secondary endpoints included liver function, postoperative complications, mortality, and duration of hospital stay.RESULTS: The characteristics of the two groups were closely matched. The Pringle maneuver was not used in the RF+ group. There was significantly less median intraoperative blood loss in the RF+ group(300 vs 400 m L, P = 0.01). On postoperative days(POD) 1 and 5, median alanine aminotransferase was significantly higher in the RF+ group than in the RF- group(POD 1: 348.5 vs 245.5, P = 0.01; POD 5: 112 vs 82.5, P = 0.00), but there was no significant difference between the two groups on POD 3(260 vs 220, P = 0.24). The median AST was significantly higher in the RF+ group on POD 1(446 vs 268, P = 0.00), but there was no significant difference between the two groups on POD 3 and 5(POD 3: 129.5 vs 125, P = 0.65; POD 5: 52.5vs 50, P = 0.10). Overall, the rate of postoperative complications was roughly the same in these two groups(28.0% vs 17.7%, P = 0.11) except that post hepatectomy liver failure was far more common in the RF+ group than in the RF- group(6% vs 0%, P = 0.04).CONCLUSION: Radiofrequency-assisted hepatectomy can reduce intraoperative blood loss during liver resection effectively. However, this method should be used with caution in patients with concomitant cirrhosis because it may cause severe liver damage and liver failure.

New advances in hepatocellular carcinoma

AIM: To determine if gene-specific DNA methylation in prospectively collected blood samples is associated with later development of hepatocellular carcinoma(HCC).METHODS: Comparing genome-wide DNA methylation profiles using Illumina Human methylation 450 K arrays, we previously identified a list of loci that were differentially methylated between tumor and adjacent nontumor tissues. To examine if dysregulation of DNAmethylation patterns observed in tumor tissues can be detected in white blood cell(WBC) DNA, we conducted a prospective case-control study nested within a community-based cancer screening cohort in Taiwan with 16 years of follow up. We measured methylation levels in ninety-six loci that were aberrant in DNA methylation in HCC tumor tissues compared to adjacent tissues. Baseline WBC DNA from 159 HCC cases and 312 matched controls were bisulfite treated and assayed by Illumina Bead Array. We used the χ2 test for categorical variables and student's t-test for continuous variables to assess the difference in selected characteristics between cases and controls. To estimate associations with HCC risk, we used conditional logistic regression models stratified on the matching factors to calculate odds ratios(OR) and 95%CI. RESULTS: We found that high methylation level in cg10272601 in WNK2 was associated with increased risk of HCC, with an OR of 1.91(95%CI: 1.27-2.86). High methylation levels in both cg12680131 in TPO and cg22511877 in MYT1 L, however, were associated with decreased risk. The ORs(95%CI) were 0.59(0.39-0.87) and 0.50(0.33-0.77), respectively, for those with methylation levels of cg12680131 and cg22511877 above the median compared with those with levels below the median. These associations were still statistically significant in multivariable conditional logistic regression models after adjusting for hepatitis B virus infection and alcohol consumption. CONCLUSION: These findings support the measurement of methylation markers in WBC DNA as biomarkers of HCC susceptibility but should be replicated in additional prospective studies.

Transforming growth factor-β and peripheral regulatory cells are negatively correlated with the overall survival of hepatocellular carcinoma

AIM To understand the cellular and molecular changes inperipheral blood that can lead to the development of hepatocellular carcinoma(HCC) and provide new methods for its diagnosis and treatment.METHODS Peripheral blood mononuclear cells were isolated from the peripheral blood of HCC patients and normal controls and then analyzed by flow cytometry. The percentage of transforming growth factor-β(TGF-β)+ regulatory cells(Tregs) in the peripheral blood was measured, and the expression of TGF-β was also determined. Then, the relationship between the changes and the 5-year survival of patients was analyzed. In addition, recombinant human TGF-β(rh TGF-β) and recombinant human interleukin-6 were added to stimulate the cultured cells, and their effects on HCC were evaluated.RESULTS The expression of TGF-β and the percentage of TGF-β+ Tregs in the peripheral blood of HCC patients increased significantly compared with normal controls. Compared with the low TGF-β expression group, the high TGF-β expression group had a significantly lower 5-year survival rate, and the same result was found in the two TGF-β+ Treg groups, suggesting that TGF-β and TGF-β+ Tregs were negatively correlated with the overall survival of the patients. In addition, rh TGF-β promoted the growth of tumor cells and induced high expression levels of IL-6, which further promoted tumor proliferation.CONCLUSION The results showed that TGF-β may promote tumor growth and proliferation by inducing the production of IL-6, and TGF-β and TGF-β+ Tregs may serve as new markers for predicting a poor prognosis in HCC.

Thrombocytopenia for prediction of hepatocellular carcinoma recurrence: Systematic review and meta-analysis

AIM: To investigate the association between thrombocytopenia and relapse after treatment for hepatocellular carcinoma(HCC).METHODS: We searched the Pub Med, EMBASE, and Web of Science databases to obtain eligible studies. The hazard ratios(HRs) values and 95% confidence intervals(CIs) were pooled by random effects model. Subsequently, we estimated the heterogeneity, performed a sensitivity analysis, determined the publication bias, and performed subgroup and meta-regression analyses. Study quality was assessed by using the Oxford Center for Evidence Based Medicine tool.RESULTS: We identified 18 eligible studies by retrieval(published during 2000-2014). Out of the 4163 patients with HCC who were recruited, 2746(66.0%) experienced recurrence. In general, our meta-analysis suggested that low platelet count(PLT) before therapy significantly increased the probability of postoperative recurrence(HR = 1.53, 95%CI: 1.29-1.81). PLT was also valuable in the prediction of intrahepatic distant recurrence(HR = 1.49, 95%CI: 1.25-1.77). Subgroupand meta-regression analyses identified various therapeutic modalities as the source of a high degree of heterogeneity. The pooled HR values showed no obvious change when a single study was removed, but otherwise, an opposite-effects model was used. In addition, no significant publication bias was detected.CONCLUSION: Thrombocytopenia before treatment might be an inexpensive and useful predictor of postoperative recurrence in patients with HCC.