PTEN status determines therapeutic vulnerability to celastrol in cholangiocarcinoma.

BACKGROUND: A balanced protein homeostasis network helps cholangiocarcinoma (CCA) maintain their oncogenic growth, and disrupting proteostasis therapeutically will induce proteotoxic stress. Phosphatase and tensin homolog (PTEN) have been reported to be involved in proteostasis, and PTEN-associated pathways are commonly altered in CCA. Celastrol, a triterpene from plants, exhibits cytotoxic effects in various types of cancer. However, the underlying mechanisms remain unclear. PURPOSE: We investigated the therapeutic effect of celastrol in CCA and identified the molecular characteristics of tumors that were sensitive to celastrol. The target of celastrol was explored. We then evaluated the candidate combination therapeutic strategy to increase the effectiveness of celastrol in celastrol-insensitive CCA tumors. METHODS: Various CCA cells were categorized as either celastrol-sensitive or celastrol-insensitive based on their response to celastrol. The molecular characteristics of cells from different groups were determined by RNA-seq. PTEN status and its role in proteasome activity in CCA cells were investigated. The CMAP analysis, molecular docking, and functional assay were performed to explore the effect of celastrol on proteasome activities. The correlation between PTEN status and clinical outcomes, as well as proteasomal activity, were measured in CCA patients. The synergistic therapeutic effect of autophagy inhibitors on celastrol-insensitive CCA cells were measured. RESULTS: Diverse responses to celastrol were observed in CCA cells. PTEN expression varied among different CCA cells, and its status could impact cell sensitivity to celastrol: PTENhigh tumor cells were resistant to celastrol, while PTENlow cells were more sensitive. Celastrol induced proteasomal dysregulation in CCA cells by directly targeting PSMB5. Cells with low PTEN status transcriptionally promoted proteasome subunit expression in an AKT-dependent manner, making these cells more reliant on proteasomal activities to maintain proteostasis. This caused the PTENlow CCA cells sensitive to celastrol. A negative correlation was found between PTEN levels and the proteasome signature in CCA patients. Moreover, celastrol treatment could induce autophagy in PTENhigh CCA cells. Disrupting the autophagic pathway in PTENhigh CCA cells enhanced the cytotoxic effect of celastrol. CONCLUSION: PTEN status in CCA cells determines their sensitivity to celastrol, and autophagy inhibitors could enhance the anti-tumor effect in PTENhigh CCA.

Mechanism exploration of Zoledronic acid combined with PD-1 in the treatment of hepatocellular carcinoma.

How to increase the response of immune checkpoint inhibitors (ICIs) is a challenge. In clinical, we found that Zoledronic acid (ZA) may increase the anti-tumor effect of immunotherapy for hepatocellular carcinoma (HCC). To explore the underlying mechanism, we established a mouse model of HCC by subcutaneously injecting Hepa1-6 cell line. The result showed that the tumor volume in the ZA plus anti-PD-1 monocloning antibody (anti-PD-1 mAb) treatment groups was significantly smaller than that of control group, and the onset time of tumor inhibition was even shorter than that of the anti-PD-1 mAb group. Using flow cytometry (FC) to detect the proportion of major immune cell subsets in tumor tissues of each group of mice, we found that the synergistic anti-tumor effect of ZA and anti-PD-1 mAb may be related to ZA-induced polarization of macrophages toward the M1 phenotype. Next, we performed bulk RNA sequencing on tumor samples from different groups to obtain differentially expressed genes (DEGs), which were then input DEGs into pathway enrichment analysis. Data indicated that ZA participated in the M1-type polarization via ferroptosis-related pathways. Our results revealed how ZA involves in the anti-tumor effect of PD-1 monoclonal antibody and provided a potential therapeutic candidate for patients with HCC.

Pharmacokinetic Study of Ultrasmall Superparamagnetic Iron Oxide Nanoparticles HY-088 in Rats.

BACKGROUND AND OBJECTIVE: HY-088 injection is an ultrasmall superparamagnetic iron oxide nanoparticle (USPIOs) composed of iron oxide crystals coated with polyacrylic acid (PAA) on the surface. The purpose of this study was to investigate the pharmacokinetics, tissue distribution, and mass balance of HY-088 injection. METHODS: The pharmacokinetics of [55Fe]-HY-088 and [14C]-HY-088 were investigated in 48 SD rats by intravenous injection of 8.5 (low-dose group), 25.5 (medium-dose group), and 85 (high-dose group) mg/100 µCi/kg. Tissue distribution was studied by intravenous injection of 35 mg/100 µCi/kg in 48 SD rats, and its tissue distribution in vivo was obtained by ex vivo tissue assay. At the same time, [14C]-HY-088 was injected intravenously at a dose of 25.5 mg/100 µCi/kg into 16 SD rats, and its tissue distribution in vivo was studied by quantitative whole-body autoradiography. [14C]-HY-088 and [55Fe]-HY-088 were injected intravenously into 24 SD rats at a dose of 35 mg/100 µCi/kg, and their metabolism was observed. RESULTS: In the pharmacokinetic study, [55Fe]-HY-088 reached the maximum observed concentration (Cmax) at 0.08 h in the low- and medium-dose groups of SD rats. [14C]-HY-088 reached Cmax at 0.08 h in the three groups of SD rats. The area under the concentration-time curve (AUC) of [55Fe]-HY-088 and [14C]-HY-088 increased with increasing dose. In the tissue distribution study, [55Fe]-HY-088 and [14C]-HY-088 were primarily distributed in the liver, spleen, and lymph nodes of both female and male rats. In the mass balance study conducted over 57 days, the radioactive content of 55Fe from [55Fe]-HY-088 was primarily found in the carcass, accounting for 86.42 ± 4.18% in females and 95.46 ± 6.42% in males. The radioactive recovery rates of [14C]-HY-088 in the urine of female and male rats were 52.99 ± 5.48% and 60.66 ± 2.23%, respectively. CONCLUSIONS: Following single intravenous administration of [55Fe]-HY-088 and [14C]-HY-088 in SD rats, rapid absorption was observed. Both [55Fe]-HY-088 and [14C]-HY-088 were primarily distributed in the liver, spleen, and lymph nodes. During metabolism, the radioactivity of [55Fe]-HY-088 is mainly present in the carcass, whereas the 14C-labeled [14C]-HY-088 shell PAA is eliminated from the body mainly through the urine.

Cell-free DNA testing for early hepatocellular carcinoma surveillance.

BACKGROUND: Liver cirrhosis (LC) is the highest risk factor for hepatocellular carcinoma (HCC) development worldwide. The efficacy of the guideline-recommended surveillance methods for patients with LC remains unpromising. METHODS: A total of 4367 LCs not previously known to have HCC and 510 HCCs from 16 hospitals across 11 provinces of China were recruited in this multi-center, large-scale, cross-sectional study. Participants were divided into Stage Ⅰ cohort (510 HCCs and 2074 LCs) and Stage Ⅱ cohort (2293 LCs) according to their enrollment time and underwent Tri-phasic CT/enhanced MRI, US, AFP, and cell-free DNA (cfDNA). A screening model called PreCar Score was established based on five features of cfDNA using Stage Ⅰ cohort. Surveillance performance of PreCar Score alone or in combination with US/AFP was evaluated in Stage Ⅱ cohort. FINDINGS: PreCar Score showed a significantly higher sensitivity for the detection of early/very early HCC (Barcelona stage A/0) in contrast to US (sensitivity of 51.32% [95% CI: 39.66%-62.84%] at 95.53% [95% CI: 94.62%-96.38%] specificity for PreCar Score; sensitivity of 23.68% [95% CI: 14.99%-35.07%] at 99.37% [95% CI: 98.91%-99.64%] specificity for US) (P < 0.01, Fisher's exact test). PreCar Score plus US further achieved a higher sensitivity of 60.53% at 95.08% specificity for early/very early HCC screening. INTERPRETATION: Our study developed and validated a cfDNA-based screening tool (PreCar Score) for HCC in cohorts at high risk. The combination of PreCar Score and US can serve as a promising and practical strategy for routine HCC care. FUNDING: A full list of funding bodies that contributed to this study can be found in Acknowledgments section.

Simultaneous nitrate and sulfate biotransformation driven by different substrates: comparison of carbon sources and metabolic pathways at different C/N ratios.

Nitrate (NO3-) and sulfate (SO42-) often coexist in organic wastewater. The effects of different substrates on NO3- and SO42- biotransformation pathways at various C/N ratios were investigated in this study. This study used an activated sludge process for simultaneous desulfurization and denitrification in an integrated sequencing batch bioreactor. The results revealed that the most complete removals of NO3- and SO42- were achieved at a C/N ratio of 5 in integrated simultaneous desulfurization and denitrification (ISDD). Reactor Rb (sodium succinate) displayed a higher SO42- removal efficiency (93.79%) with lower chemical oxygen demand (COD) consumption (85.72%) than reactor Ra (sodium acetate) on account of almost 100% removal of NO3- in both Ra and Rb. Ra produced more S2- (5.96 mg L-1) and H2S (25 mg L-1) than Rb, which regulated the biotransformation of NO3- from denitrification to dissimilatory nitrate reduction to ammonium (DNRA), whereas almost no H2S accumulated in Rb which can avoid secondary pollution. Sodium acetate-supported systems were found to favor the growth of DNRA bacteria (Desulfovibrio); although denitrifying bacteria (DNB) and sulfate-reducing bacteria (SRB) were found to co-exist in both systems, Rb has a greater keystone taxa diversity. Furthermore, the potential carbon metabolic pathways of the two carbon sources have been predicted. Both succinate and acetate could be generated in reactor Rb through the citrate cycle and the acetyl-CoA pathway. The high prevalence of four-carbon metabolism in Ra suggests that the carbon metabolism of sodium acetate is significantly improved at a C/N ratio of 5. This study has clarified the biotransformation mechanisms of NO3- and SO42- in the presence of different substrates and the potential carbon metabolism pathway, which is expected to provide new ideas for the simultaneous removal of NO3- and SO42- from different media.

Aquaporin 1 is a prognostic marker and inhibits tumour progression through downregulation of Snail expression in intrahepatic cholangiocarcinoma.

BACKGROUND: Recently, some studies have suggested a link between AQP1 and cancer progression. AIMS: The aim of the present study was to investigate the influence of AQP1 on the clinicopathology and prognosis of intrahepatic cholangiocarcinoma (ICC) patients. METHODS: We retrospectively detected the expression of AQP1 protein in 307 patients with ICC who underwent partial hepatectomy. Western blot analysis was used to detect AQP1 protein levels in stable AQP1 overexpression and knockdown cell lines. The influence of AQP1 on the invasion and metastasis ability of ICC cells was assessed by wound-healing and Transwell assays in vitro as well as by a splenic liver metastasis model in vivo. RESULTS: Positive membranous AQP1 expression was identified in 34.2% (105/307) of the ICC specimens. Survival data revealed that positive AQP1 expression was significantly associated with favourable disease-free survival (DFS) and overall survival (OS) (p = 0.0290 and p = 0003, respectively). Moreover, high AQP1 expression inhibited the invasion and migration of ICC cells in vitro as well as inhibited liver metastasis in nude mice. Mechanistically, high AQP1 expression in ICC cells increased the levels of E-cadherin but decreased the levels of the Snail transcription factor. CONCLUSIONS: AQP1 expression is associated with a favourable prognosis in ICC patients. AQP1 inhibits ICC cell invasion, metastasis, and epithelial-mesenchymal transition (EMT) through downregulation of Snail expression.

The development of a cSMART-based integrated model for hepatocellular carcinoma diagnosis.

BACKGROUND: Hepatocellular carcinoma (HCC) generally arises from a background of liver cirrhosis (LC). Patients with cirrhosis and suspected HCC are recommended to undergo serum biomarker tests and imaging diagnostic evaluation. However, the performance of routine diagnostic methods in detecting early HCC remains unpromising. METHODS: Here, we conducted a large-scale, multicenter study of 1675 participants including 490 healthy controls, 577 LC patients, and 608 HCC patients from nine clinical centers across nine provinces of China, profiled gene mutation signatures of cell-free DNA (cfDNA) using Circulating Single-Molecule Amplification and Resequencing Technology (cSMART) through detecting 931 mutation sites across 21 genes. RESULTS: An integrated diagnostic model called "Combined method" was developed by combining three mutation sites and three serum biomarkers. Combined method outperformed AFP in the diagnosis of HCC, especially early HCC, with sensitivities of 81.25% for all stages and 66.67% for early HCC, respectively. Importantly, the integrated model exhibited high accuracy in differentiating AFP-negative, AFP-L3-negative, and PIVKA-II-negative HCCs from LCs.

Expert Consensus on Pathological Diagnosis of Intrahepatic Cholangiocarcinoma (2022 version).

Intrahepatic cholangiocarcinoma (iCCA) can originate from the large bile duct group (segment bile ducts and area bile ducts), small bile duct group (septal bile ducts and interlobular bile ducts), and terminal bile duct group (bile ductules and canals of Hering) of the intrahepatic biliary tree, which can be histopathological corresponding to large duct type iCCA, small duct type iCCA and iCCA with ductal plate malformation pattern, and cholangiolocarcinoma, respectively. The challenge in pathological diagnosis of above subtypes of iCCA falls in the distinction of cellular morphologies, tissue structures, growth patterns, invasive behaviors, immunophenotypes, molecular mutations, and surgical prognoses. For these reasons, this expert consensus provides nine recommendations as a reference for standardizing and refining the diagnosis of pathological subtypes of iCCA, mainly based on the 5th edition of the World Health Organization Classification of Tumours of the Digestive System.

Kitchen-waste-derived biochar modified nanocomposites with improved photocatalytic performances for degrading organic contaminants.

Kitchen-waste-derived biochar (KBC) was produced by thermal treatment at 400 °C, and a series of KBC/BiOX (X = Br, Cl) photocatalysts were developed using ultrasonication and solvothermal treatment. The as-prepared photocatalysts were characterized by several tests and investigated by photocatalytic reactions towards methyl orange (MO) and tetracycline (TC). The best photocatalysts, 0.15KBC/BiOBr and 0.15KBC/BiOCl separately achieved complete MO photodegradation in 20 min and 35 min. Further study confirmed that 0.15KBC/BiOBr and 0.15KBC/BiOCl possessed excellent photocatalytic efficiency that was 17.9 and 14.8 times higher than BiOBr and BiOCl, respectively. In addition, 0.15KBC/BiOX showed higher activity removal of TC than pure BiOX in 60 min. Notably, 0.15KBC/BiOX maintained a reproducible high photocatalytic efficiency after five recycles. Estimated band gap energy for 0.15KBC/BiOBr (2.40 eV) and 0.15KBC/BiOCl (3.00 eV) was considerably lower than that of BiOBr (2.73 eV) and BiOCl (3.30 eV), indicating a delocalized state was created when forming electronic pathways on the interface. Besides, visible-light harvesting of photocatalysts got promoted by the modification of KBC. Active species trapping experiments and electron paramagnetic resonance (EPR) tests illustrated that photogenerated holes were the principal active species, while ∙OH was involved in the reaction. The successful synthesis of 0.15KBC/BiOX catalyst provided a new approach on simultaneously degrading organic contaminants in water and disposing of excessive kitchen waste.

Evaluation of artificial intelligence models for the detection of asymmetric keratoconus eyes using Scheimpflug tomography.

BACKGROUND: To evaluate artificial intelligence (AI) models based on objective indices and raw corneal data from the Scheimpflug Pentacam HR system (OCULUS Optikgeräte GmbH, Wetzlar, Germany) for the detection of clinically unaffected eyes in patients with asymmetric keratoconus (AKC) eyes. METHODS: A total of 1108 eyes of 1108 patients were enrolled, including 430 eyes from normal control subjects, 231 clinically unaffected eyes from patients with AKC, and 447 eyes from keratoconus (KC) patients. Eyes were divided into a training set (664 eyes), a test set (222 eyes) and a validation set (222 eyes). AI models were built based on objective indices (XGBoost, LGBM, LR and RF) and entire corneal raw data (KerNet). The discriminating performances of the AI models were evaluated by accuracy and the area under the ROC curve (AUC). RESULTS: The KerNet model showed great overall discriminating power in the test (accuracy = 94.67%, AUC = 0.985) and validation (accuracy = 94.12%, AUC = 0.990) sets, which were higher than the index-derived AI models (accuracy = 84.02%-86.98%, AUC = 0.944-0.968). In the test set, the KerNet model demonstrated good diagnostic power for the AKC group (accuracy = 95.24%, AUC = 0.984). The validation set also proved that the KerNet model was useful for AKC group diagnosis (accuracy = 94.12%, AUC = 0.983). CONCLUSIONS: KerNet outperformed all the index-derived AI models. Based on the raw data of the entire cornea, KerNet was helpful for distinguishing clinically unaffected eyes in patients with AKC from normal eyes.

A Corresponding Region Fusion Framework for Multi-modal Cervical Lesion Detection.

Cervical lesion detection (CLD) using colposcopic images of multi-modality (acetic and iodine) is critical to computer-aided diagnosis (CAD) systems for accurate, objective, and comprehensive cervical cancer screening. To robustly capture lesion features and conform with clinical diagnosis practice, we propose a novel corresponding region fusion network (CRFNet) for multi-modal CLD. CRFNet first extracts feature maps and generates proposals for each modality, then performs proposal shifting to obtain corresponding regions under large position shifts between modalities, and finally fuses those region features with a new corresponding channel attention to detect lesion regions on both modalities. To evaluate CRFNet, we build a large multi-modal colposcopic image dataset collected from our collaborative hospital. We show that our proposed CRFNet surpasses known single-modal and multi-modal CLD methods and achieves state-of-the-art performance, especially in terms of Average Precision.

Construction of mannose-modified polyethyleneimine-block-polycaprolactone cationic polymer micelles and its application in acute lung injury.

This study evaluated the D-mannose modified polyethyleneimine-block-polycaprolactone biomacromolecule copolymer micelles (PCL-PEI-mannose) as a targeted delivery of the glucocorticoid dexamethasone (DXM) to lung inflammation tissues and enhances the vehicle for its anti-inflammatory effects. Dexamethasone was encapsulated in the hydrophobic core of cationic polymer micelles by solvent evaporation. The polymeric micelles exhibited sustained-release within 48 h, good blood compatibility, and colloidal stability in vitro. The cellular uptake of mannose-modified micelles was higher compared with the non-modified micelles. And drug-loaded targeted micelles could inhibit the production of inflammatory factors in activated RAW264.7 cells. The distribution results indicated that drug-loaded targeted micelles highly improved the lung targeting ability, reduced the wet/dry ratio of injured lung tissue, and relieved the lung inflammation, accompanied by the decrease of inflammatory cell infiltration, myeloperoxidase activity, and inflammatory mediator levels in bronchoalveolar lavage fluid. These findings suggested that PCL-PEI-mannose delivery system could facilitate the lung-specific delivery and inhibit the inflammatory response. Collectively, PCL-PEI-mannose polymer micelles could be used as a potential delivery system for the treatment of acute lung injury (ALI).

Chinese Expert Consensus on Immunotherapy for Hepatocellular Carcinoma (2021 Edition).

Background: Hepatocellular carcinoma (HCC) is one of the most common malignancies in China. Most HCC patients are first diagnosed at an advanced stage, and systemic treatments are the mainstay of treatment. Summary: In recent years, immune checkpoint inhibitors have made a breakthrough in the systemic treatment of middle-advanced HCC, breaking the single therapeutic pattern of molecular-targeted agents. To better guide the clinical treatment for effective and safe use of immunotherapeutic drugs, the Chinese Association of Liver Cancer and Chinese Medical Doctor Association has gathered multidisciplinary experts and scholars in relevant fields to formulate the "Chinese Clinical Expert Consensus on Immunotherapy for Hepatocellular Carcinoma (2021)" based on current clinical studies and clinical medication experience for reference in China. Key Messages: The consensus contained 17 recommendations, including the preferred regimen for first- and second-line immunotherapy, evaluation and monitoring before/during/after treatment, management of complications, precautions for special patients, and potential population for immunotherapy.

Study on the effect of the size irregularity gradient of metal foams on macroscopic compressive properties.

Size irregularity gradient and cell wall gradient, combined as the density gradient in previous studies, affect the macroscopic mechanical properties of the gradient metal foam. More and more complex mesostructures are designed and applied in metal foams, and the density gradient becomes insufficient to describe the difference in mesostructures. To explore the effect of mesostructures carefully, this study focuses on the effect of the size irregularity gradient on the macroscopic compressive properties of metal foams. A series of metal foam models were developed using the 3D Voronoi technique. These models have the same average relative densities, the same average diameters and different size irregularity gradients. Simulation results indicated that the macroscopic mechanical properties of cell wall gradient metal foams are significantly different from those of size irregularity gradient metal foams, though these models have the same relative density gradient. To explore the effect of size irregularity gradient, a theoretical model was developed to characterize the compression process from the first cell-collapse to full condensation. Theoretical results showed a linear relationship between the nominal stress and the current relative density. These findings can provide efficient guidance for the design and applications of gradient metal foams.

RPRD1A stabilizes NRF2 and aggravates HCC progression through competing with p62 for TRIM21 binding.

NRF2 is the master transcriptional activator of cytoprotective genes and Kelch-like ECH-associated protein 1 (Keap1), a biosensor for electrophiles and oxidation, promotes NRF2 degradation in unstressed conditions. SQSTM1/p62, an oncogenic protein aberrantly accumulated in hepatocellular carcinoma (HCC), binds and sequestrates Keap1, leading to the prevention of NRF2 degradation. Here, we show that p15INK4b-related sequence/regulation of nuclear pre-mRNA domain-containing protein 1A (RPRD1A) is highly expressed in HCC tumors and correlated with aggressive clinicopathological features. RPRD1A competitively interacts with TRIM21, an E3 ubiquitin ligase of p62, resulting in the decrease of p62 ubiquitination and the increased sequestration for Keap1. Therefore, RPRD1A enhances the nuclear translocation of NRF2, which induces gene expression for counteracting oxidative stress, maintaining cancer cells survival, and promoting HCC development. Moreover, disturbing the redox homeostasis of cancer cells by genetic knockdown of RPRD1A sensitizes cancer cells to platinum-induced cell death. Our study reveals RPRD1A is involved in the oxidative stress defense program and highlights the therapeutic benefits of targeting pathways that support antioxidation.

Comparison of Modified Child-pugh (MCP), Albumin-bilirubin (ALBI), and Child-pugh (CP) score for predicting of survival in Hepatocellular Carcinoma Patients Treated with Transcatheter Arterial Chemoembolization.

BACKGROUND: Both modified Child-Pugh (MCP) and Albumin-bilirubin (ALBI) grade were reported that simpler, more objective and evidence-based alternative to the Child-Pugh (CP) class for assessing liver function. AIMS: To investigate whether the MCP and ALBI grade could better evaluate the liver reserve of Hepatocellular Carcinoma (HCC) patients treated with TACE (transcatheter arterial chemoembolization) than CP grade. METHODS: Three hundred seventy-six consecutive HCC patients treated with TACE between December 2007 and October 2011 were enrolled. The baseline characteristics and clinical information were collected. Homogeneity and discriminatory ability were compared between the MCP grade and ALBI class or CP grade. RESULTS: Compared with the CP and ALBI, the MCP grade had a higher predictive accuracy for overall survival (OS) in terms of homogeneity and discriminatory ability. Most of the HCC patients had CP class A disease (84.0%) at presentation, and within this CP class, although the ALBI grade revealed two clear and nonoverlapping groups, the MCP grade revealed three clearly different prognostic groups. Both in the ALBI grade 1 or ALBI grade 2 group, the MCP grade still showed a significant progressive decrease in OS from the smallest to the largest grades, but the CP class was unsatisfactory in stratifying these patients. CONCLUSIONS: The stratification ability and prognostic predictive power of the MCP grade for HCC patients treated with TACE may be better than that of the ALBI grade or CP class.

The Landscape of Cell-Free HBV Integrations and Mutations in Cirrhosis and Hepatocellular Carcinoma Patients.

PURPOSE: Intratumoral hepatitis B virus (HBV) integrations and mutations are related to hepatocellular carcinoma (HCC) progression. Circulating cell-free DNA (cfDNA) has shown itself as a powerful noninvasive biomarker for cancer. However, the HBV integration and mutation landscape on cfDNA remains unclear. EXPERIMENTAL DESIGN: A cSMART (Circulating Single-Molecule Amplification and Resequencing Technology)-based method (SIM) was developed to simultaneously investigate HBV integration and mutation landscapes on cfDNA with HBV-specific primers covering the whole HBV genome. Patients with HCC (n = 481) and liver cirrhosis (LC; n = 517) were recruited in the study. RESULTS: A total of 6,861 integration breakpoints including TERT and KMT2B were discovered in HCC cfDNA, more than in LC. The concentration of circulating tumor DNA (ctDNA) was positively correlated with the detection rate of these integration hotspots and total HBV integration events in cfDNA. To track the origin of HBV integrations in cfDNA, whole-genome sequencing (WGS) was performed on their paired tumor tissues. The paired comparison of WGS data from tumor tissues and SIM data from cfDNA confirmed most recurrent integration events in cfDNA originated from tumor tissue. The mutational landscape across the whole HBV genome was first generated for both HBV genotype C and B. A region from nt1100 to nt1500 containing multiple HCC risk mutation sites (OR > 1) was identified as a potential HCC-related mutational hot zone. CONCLUSIONS: Our study provides an in-depth delineation of HBV integration/mutation landscapes at cfDNA level and did a comparative analysis with their paired tissues. These findings shed light on the possibilities of noninvasive detection of virus insertion/mutation.

KerNet: A Novel Deep Learning Approach for Keratoconus and Sub-Clinical Keratoconus Detection Based on Raw Data of the Pentacam HR System.

Keratoconus is one of the most severe corneal diseases, which is difficult to detect at the early stage (i.e., sub-clinical keratoconus) and possibly results in vision loss. In this paper, we propose a novel end-to-end deep learning approach, called KerNet, which processes the raw data of the Pentacam HR system (consisting of five numerical matrices) to detect keratoconus and sub-clinical keratoconus. Specifically, we propose a novel convolutional neural network, called KerNet, containing five branches as the backbone with a multi-level fusion architecture. The five branches receive five matrices separately and capture effectively the features of different matrices by several cascaded residual blocks. The multi-level fusion architecture (i.e., low-level fusion and high-level fusion) moderately takes into account the correlation among five slices and fuses the extracted features for better prediction. Experimental results show that: (1) our novel approach outperforms state-of-the-art methods on an in-house dataset, by ~1% for keratoconus detection accuracy and ~4 for sub-clinical keratoconus detection accuracy; (2) the attention maps visualized by Grad-CAM show that our KerNet places more attention on the inferior temporal part for sub-clinical keratoconus, which has been proved as the identifying regions for ophthalmologists to detect sub-clinical keratoconus in previous clinical studies. To our best knowledge, we are the first to propose an end-to-end deep learning approach utilizing raw data obtained by the Pentacam HR system for keratoconus and subclinical keratoconus detection. Further, the prediction performance and the clinical significance of our KerNet are well evaluated and proved by two clinical experts. Our code is available at https://github.com/upzheng/Keratoconus.

Influence of postoperative adjuvant transarterial chemoembolization on the prognosis of early-stage intrahepatic cholangiocarcinoma: a single center study.

BACKGROUND: The effectiveness of postoperative adjuvant transarterial chemoembolization (TACE) on survival and recurrence in tumor-node-metastasis (TNM) stage I intrahepatic cholangiocarcinoma (ICC) after radical resection remains unclear. This study aimed to compare overall survival (OS) and recurrence-free survival (RFS) in TNM stage I ICC patients with and without postoperative TACE. METHODS: A retrospective cohort study was conducted on TNM stage I ICC patients who had undergone R0 resections with curative intent in Shanghai Eastern Hepatobiliary Surgery Hospital from January 2012 to December 2016. A total of 269 patients were divided into two groups: (I) 35 patients who received postoperative TACE and (II) 234 patients no TACE. Staging was performed according to the 8th edition of American Joint Committee on Cancer (AJCC) TNM staging system. The tumor-related RFS and OS were estimated by the Kaplan-Meier method. Cox proportional regression model was employed to evaluate the prognosis between the two groups. RESULTS: In all patients, the median OS was 66.8 months. After R0 resection, adjuvant TACE could not improve the survival of TNM stage I patients, and the OS of the TACE group was not better than that of the non-TACE group (P=0.7070). In addition, in the TACE group, the recurrence rate of TNM stage I ICC patients was statistically significantly higher than that of the non-TACE group (P=0.0328). Multivariable analysis revealed that adjuvant TACE was an independent predictor of worse RFS (HR: 1.88, 95% CI: 1.21-2.93). CONCLUSIONS: Adjuvant TACE after radical surgery failed to prolong the OS and potentially delay recurrence for patients with TNM Stage I ICC. Adjuvant TACE might not be suitable for patients with TNM Stage I ICC.

MiR-371 promotes proliferation and metastasis in hepatocellular carcinoma by targeting PTEN.

Hepatocellular carcinoma (HCC) is the leading cause of cancer-related mortality worldwide. MiR-371 has recently emerged as an important regulator in tumorigenesis, and may serve as a biomarker for malignant tumors. We transfected miR-371 or its inhibitor in two human HCC cell lines, then used 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, soft agar colony formation, and transwell migration assays to evaluate the effects on cell proliferation, migration, and invasion. We found that miR-371 was positively correlated with HCC metastasis and poor prognosis in the inflicted patients, and the high expression of miR-371 was promoted, whereas a low level of miR-371 depressed cell proliferation and invasion. We found PTEN to be a direct target of miR-371. The overexpression or knockdown of PTEN exhibited the opposite effects from those of miR-371 on cell proliferation and migration. Our study demonstrates that miR-371 promotes proliferation and metastasis in HCC by targeting PTEN. [BMB Reports 2019; 52(5): 312-317].