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BACKGROUND: Many studies have indicated that autophagy plays an important role in multiple cancers, including hepatocellular carcinoma (HCC). This study aimed to establish a prognostic signature for HCC based on autophagy-related genes (ARGs) to predict the prognosis of patients. METHODS: The list of ARGs was derived from screening National Center for Biotechnology Information (NCBI)-Gene and Molecular Signatures Database (MSigDB) datasets. Differential analysis was conducted via the R limma package in HCC patients based on The Cancer Genome Atlas (TCGA) database. Univariate and multivariate Cox regression analysis were conducted to identify key prognostic ARGs via the survival package. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed by clusterProfiler package. The Estimation of Stromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) algorithm was used to conduct immune analysis. Finally, the correlation between the prognostic model and clinical characteristics was also assessed, including age, tumor-node-metastasis (TNM) stages, and tumor grades. RESULTS: Firstly, 106 differential ARGs were identified and 10 candidates were further confirmed via Cox regression analysis, including BAMBI, HIF1A, SERPINE1, EZH2, SLC9A3R1, IGFBP3, HSPB8, DAB2, CXCL1 and PRNP. The receiver operating characteristic (ROC) curve analysis revealed that the ARGs risk model had a well diagnostic positive rate with 1-year area under the curve (AUC) =0.688 and 3-year AUC =0.674. Correlation analysis indicated that only advanced tumor stages were positively associated with high ARGs scores with P=0.0227. There were also significant differences in tumor purity (P=6.71e-05), infiltrating cell analysis (P=7.77e-05), immune analysis (P=7.9e-05), and stromal cells analysis (P=0.0015) in high- and low-risk ARGs samples. The genes HIF1A, IGFBP3, and DAB2 were found to have high frequent missense mutations in samples with high-risk ARGs scores. Lastly, we also established a nomogram to predict overall survival (OS) of HCC by integrating ARGs scores and other clinical parameters. CONCLUSIONS: Our study established an autophagy-related signature for predicting the prognosis of HCC patients, providing a thorough understanding of the underlying mechanisms of autophagy in HCC.

Patients with persistent positive hepatitis B surface antigen (HBsAg), even with a low HBV-DNA load, have a higher risk of hepatocellular carcinoma (HCC) than those without HBV infection. Given that tumor stemness has a critical role in the occurrence and maintenance of neoplasms, this study aimed to explore whether HBsAg affects biological function and stemness of HCC by regulating microRNA, and to explore underlying mechanisms.We screened out miR-203a, the most significant down-regulated microRNA in the microarray analysis of HBsAg-positive samples and focused on that miRNA in the ensuing study.MiR-203a was significantly down-regulated in HBsAg-positive HCC with the sharpest decrease shown in microarray analysis. The negative correlation between miR-203a and HBsAg expression was confirmed by quantitative real-time PCR after stimulation or overexpression/knockdown of HBsAg in cells. We demonstrated the function of miR-203a in inhibiting HCC cell proliferation, migration, clonogenic capacity, and tumor developmentHBsAg may promote the development of HCC and tumor stemness by inhibiting miR-203a, resulting in poor prognosis. miR-203a may serve as a crucial treatment target in HBsAg-positive HCC. More explicit mechanistic studies and animal experiments need to be conducted as a next step.

Magneto-Acousto-Electrical Tomography (MAET) is a novel multi-physics imaging method, which promises to offer a unique biophysical property of tissue electrical impedance with the additional benefit of excellent spatial resolution of the ultrasonic imaging. It opens the potential for early diagnosis of cancer by revealing changes of dielectric characteristics. However, direct MAET is unable to image the irregularly-shaped lesions fully due to the dependence on the angle between conductivity boundary and ultrasound beam direction. In this paper, a numerical simulation of multi-angle MAET is presented for an improved image reconstruction for MAET in order to discern irregularly-shaped tumors in different positions. The results show that the conductivity boundary interfaces are invisible in single angle B-mode reconstructed image, wherever the ultrasound beam and conductivity boundary are nearly parallel. When the multi-angle scanning was adopted, the image reconstructed with image rotation method reproduced the original object pattern. Furthermore, the relationship between reconstruction error and the number of angles was also discussed. It is found that 12 angles would be necessary to achieve nearly the optimal reconstruction. Finally, reconstructed images in L2 norm of the error with the measurement noise are presented.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease. The noninvasive and accurate classification of NAFLD is still a challenging problem. In this study we pro- posed a new quantitative ultrasound (QUS) technique, which combined multiple QUS parameters for distinguishing steatosis stages. NAFLD was induced in the livers of 57 rats by gavage feeding with a high fat emulsion, while 8 rats were given a standard diet to serve as controls. Ex vivo ultrasound mea- surement was conducted for capturing the radiofrequency signal. Six QUS parameters were extracted and selected for linear combination. The results show that the overall performance of the combined parameter is better than that of the single QUS parameter. The accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUROC) while using our proposed method to distinguish mild steatosis (stage S1) from the steatosis under stage S0 are 90.1%, 0.93, 0.88 and 0.97 respectively. In conclusion, the proposed method in this study can make up for the deficiency of single parameter and improve the quantitative staging ability of fatty liver, and thus could play an important role in the diagnosis of NAFLD.

Immunoglobulin G4 (IgG4)-related cholecystitis (IgG4-C) is often difficult to distinguish from gallbladder carcinoma (GBC). This study aimed to determine a practical strategy for differentiating between IgG4-C and GBC to avoid unnecessary surgical resection. The expression of IgG4 in the gallbladder was detected by immunohistochemistry. The clinicopathological and radiological characteristics of IgG4-C patients and GBC patients were analyzed retrospectively. Immunohistochemistry revealed that IgG4 was upregulated in the plasma cells of IgG4-C tissues. The median serum total bilirubin levels were significantly higher in the patients with IgG4-C than in those with GBC (45.8 µmol L-1 vs. 29.9 µmol L-1). The serum γ-GGT levels were higher in IgG4-C patients than in GBC patients, whereas the serum levels of CA125 were significantly higher in GBC patients than in IgG4-C patients. The imaging scans were helpful for differentiating IgG4-C from GBC based on the presence of a layered pattern and Rokitansky-Aschoff sinuses in the gallbladder wall. There were no statistically significant differences in age, presence of abdominal pain, level of emaciation between the two groups. Our study demonstrated that the combination of imaging with serum total bilirubin, γ-GGT and CA125 levels can offer added preoperative diagnostic value and reduce the rate of IgG4-C misdiagnosis.

CA extern Dysregulation of dickkopf-related protein 1 (DKK1) expression has been reported in a variety of human cancers. We previously reported that DKK1 was upregulated in hepatocellular carcinoma (HCC). However, the role of DKK1 in HCC remains unclear. This study aimed to investigate the clinical significance and biological functions of DKK1 in HCC. The expression of DKK1 was examined in cirrhotic and HCC tissues by immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR). DKK1 was silenced or overexpressed in HCC cell lines, and in vitro and in vivo studies were performed. Immunohistochemistry revealed that DKK1 was weakly expressed in cirrhotic tissues (8/22, 36.4%) but upregulated in HCC tissues (48/53, 90.6%, cohort 1). Significant upregulation of DKK1 was observed in 57.6% (19/33, cohort 2) of HCC tissues by qRT-PCR, and the expression of DKK1 was associated with tumor size (P = 0.024) and tumor number (P = 0.019). Genetic depletion of DKK1 impaired the proliferation, colony-forming ability, invasion, and tumor formation of HCC cells (HepG2 and HUH-7). Conversely, forced expression of DKK1 increased the proliferation, colony-forming ability, and invasion of HepG2 and HUH-7 cells in vitro and enhanced tumor formation in vivo. Subsequent investigation revealed that the DKK1-mediated proliferation and tumorigenicity of HepG2 and HUH-7 cells is dependent on the Wnt/β-catenin signaling pathway. These findings indicate that DKK1 plays an oncogenic role in HCC by activating the Wnt/β-catenin signaling pathway.

The incidence of hepatocellular carcinoma (HCC), the second most frequent cause of cancer-related death worldwide, accounting for 90% of all primary liver neoplasias, is increasing.1 Recent studies have demonstrated that microRNAs (miRNAs) can act as oncogenes or tumor suppressors, and miRNA dysregulation may play an important role in cancer onset and progression.2, 3 Transcriptional and epigenetic regulations may play a major role in miRNA expression.4 However, the molecular mechanisms involved in disorders associated with miRNAs remain unclear. In recent years, miRNAs within exosomes have gained significant attention due to their potential role in delineating the molecular pathogenesis of cancer.5, 6 Exosomes are small (40-100 nm) cell-derived extracellular vesicles that initially form with the maturation of multivesicular bodies as intraluminal vesicles (ILVs).7 These ILVs are released in the extracellular medium by fusion of the limiting membrane with the plasma membrane and are thereafter known as “exosomes.”7 Cancer cells secrete exosomes that function in an autocrine or paracrine manner to modulate the tumor microenvironment.8-10 Accumulating evidence indicates that exosomes are involved in cell–cell communication by delivering proteins and nucleic acids to recipient cells, thereby impacting the biological processes in recipient cells.7, 11 However, little is known about whether exosomes can function as mediators by self-regulation of miRNA expression profiles. Cargo sorting of ILVs (exosomes) depends on the endosomal sorting complex required for transport (ESCRT) machinery.12 Loss-of-function mutations in ESCRT components lead to disorders of cargoes within ILVs, which could lead to alterations in several signaling pathways.13, 14 Membrane neck cleavage mediated by ESCRT-III is crucial for the formation of ILVs, which is regulated by Vps4.15 In humans, there are two paralogs of the yeast Vps4, Vps4A and Vps4B, both of which exhibit functions that are similar to that of the protein in yeast. Recent studies have shown that Vps4 is responsible for hepatitis B virus budding and the release of infectious hepatitis C virus particles.16, 17 Rodahl et al. reported that disruption of Vps4 and c-Jun N-terminal kinase function could cause tumor growth in Drosophila. 18 Until now, the role of Vps4 in human cancer development has been poorly understood. In this study, we demonstrated that Vps4A is frequently down-regulated in HCC tissues. Ectopic expression of Vps4A led to significant repression of HCC growth and metastasis. Our study provides mechanistic evidence that Vps4A interferes with the biological activity of exosomes and the cell response to exosomes through affecting exosomal miRNA secretion and uptake. Overexpression of Vps4A can lead to inactivation of the phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway. These results implicate Vps4A as a novel tumor suppressor for HCC.

Failure of immune surveillance related to inadequate host antitumor immune responses has been suggested as a possible cause of the high incidence of recurrence and poor overall survival outcome of hepatocellular carcinoma. The stress-induced heat shock proteins (HSPs) are known to act as endogenous "danger signals" that can improve tumor immunogenicity and induce natural killer (NK) cell responses. Exosome is a novel secretory pathway for HSPs. In our experiments, the immune regulatory effect of the HSP-bearing exosomes secreted by human hepatocellular carcinoma cells under stress conditions on NK cells was studied. ELISA results showed that the production of HSP60, HSP70, and HSP90 was up-regulated in both cell lines in a stress-specific manner. After exposure to hepatocellular carcinoma cell-resistant or sensitive anticancer drugs (hereafter referred to as "resistant" or "sensitive" anticancer drug), the membrane microvesicles were actively released by hepatocellular carcinoma cells, differing in their ability to present HSPs on the cell surface, which were characterized as exosomes. Acting as a decoy, the HSP-bearing exosomes efficiently stimulated NK cell cytotoxicity and granzyme B production, up-regulated the expression of inhibitory receptor CD94, and down-regulated the expression of activating receptors CD69, NKG2D, and NKp44. Notably, resistant anticancer drugs enhanced exosome release and generated more exosome-carried HSPs, which augmented the activation of the cytotoxic response. In summary, our findings demonstrated that exosomes derived from resistant anticancer drug-treated HepG2 cells conferred superior immunogenicity in inducing HSP-specific NK cell responses, which provided a clue for finding an efficient vaccine for hepatocellular carcinoma immunotherapy.

Background Liver enriched natural killer (NK) cells are of high immune activity. However, the function of donor liver NK cells in allogeneic liver transplantation (LTx) remains unclear. Methods Ten Gy of whole body gamma-irradiation (WBI) from a 60 Co source at 0.6 Gy/min was used for depleting donor-derived leukocytes, and transfusion of purified liver NK cells isolated from the same type rat as donor (donor type liver NK cells, dtl NKs) through portal vein was performed immediately after grafting the irradiated liver. Post-transplant survival observation on recipients and histopathological detection of liver grafts were adoptive to evaluate the biological impact of donor liver NK cells on recipients' survival in rat LTx. Results Transfusion of dtl NKs did not shorten the survival time among the recipients of spontaneous tolerance model (BN to LEW rat) after rat LTx, but prolonged the liver graft survival among the recipients depleted of donor-derived leukocytes in the acute rejection model (LEW to BN rat). Compared to the recipients in the groups which received the graft depleted of donor-derived leukocytes, better survival and less damage in the allografts were also found among the recipients in the two different strain combinations of liver allograft due to transfusion of dtl NKs. Conclusions Donor liver NK cells alone do not exacerbate liver allograft acute rejection. Conversely, they can alleviate it, and improve the recipients' survival.

AIM: To discuss strategies and prognosis for the emergency treatment of ruptured bleeding in primary hepatocellular carcinoma. METHODS: The retrospective analysis was performed by examining the emergency treatment experiences of 60 cases of ruptured bleeding in primary hepatocellular carcinoma. The treatment methods included surgical tumour resection, transcatheter arterial embolization (TAE) and non-surgical treatment. Univariate and multivariate analyses were performed to identify the risk factors that impacted 30-d mortality in the research groups. RESULTS: The 30-d mortality of all patients was 28.3% (n = 17). The univariate analysis showed that Child-Pugh C level liver function, shock, massive blood transfusion and large tumour volume were risk factors that influenced 30-d mortality. The multivariate analysis showed that shock and massive blood transfusion were independent risk factors that impacted the 30-d mortality of surgical resection. As for the TAE patients, larger tumour volume was a risk factor towards prognosis. CONCLUSION: Radical resection and TAE therapy would achieve better results in carefully selected ruptured hepatocellular tumours.

This study was to establish BALB/c murine model featured with the human-mouse chimeras from umbilical cord blood transplantation.Thirty BALB/c nude mice were exposed to 350 cGy radiation under the sublethal condition. The nuclear cells from fresh umbilical cord blood were injected into the mice of experimental group via tail vein, in which the mice were further allocated to A, B and C sub-group with given 1.0 x 10(7), 2.0 x 10(7) and 3.0 x 10(7) nuclear cells per mouse respectively, and simultaneously the equal volume normal sodium (NS) was injected into the mice of control group. White blood cells from peripheral blood in experimental group and the control group were detected at week 1, 2, 3, 4 and 8 after transplantation. Human CD34+ and CD45+ from peripheral blood in experimental group were detected by using flow cytometry analysis on week 4, 6 and 8 after transplantation in order to be the human-mouse chimeras known.No difference in the number of white blood cells showed between experimental groups and control group before blood cell transplantation and after transplantation conducted on for 8 weeks (P0.05), but the number of white blood cells from experimental groups and control group wasn't totally same on week 1, 2, 3 and 4 after cell transplantation conducted (P0.05). As compared with pre-transplantation, the number of white blood cells for post-transplantation at different time in experimental groups and control group was decreasing, and the lowest on 1 week, then going up and to the level of pre-transplantation on 4 week in experimental groups and on 8 week in control group. CD34+ and CD45+ cells in peripheral blood for nude mice appeared on 4 week, but the total number was little. The number of CD34+, CD45+ cells in experimental group A was less than those in experimental group B and C at all the stage (P0.05), but there was no difference between experimental group B and C(P0.05). There was obvious correlation between CD34+ and CD45+ on 4 week (r = 0. 903, P0.05), but this correlation didn't appear on 6 and 8 week (P0.05), which might be related to the amount of sample.The BALB/c murine model can be successfully established with the human-mouse chimeras from umbilical cord blood transplantation.