Cold Air Plasma Inhibiting Tumor-Like Biological Behavior of Rheumatoid Arthritis Fibroblast-Like Synovial Cells via G2/M Cell Cycle Arrest.

Background: Rheumatoid arthritis fibroblast-like synovial cells (RA-FLS) have become the core effector cells for the progression of rheumatoid arthritis due to their "tumor-like cell" characteristics, such as being able to break free from growth restrictions caused by contact inhibition, promoting angiogenesis, invading surrounding tissues, and leading to uncontrolled synovial growth. In recent years, cold air plasma (CAP) has been widely recognized for its clear anticancer effect. Inspired by this, this study investigated the inhibitory effect of CAP on the tumor-like biological behavior of RA-FLS through in vitro experiments. Methods: Treatment of RA-FLS with CAP at different time doses (0s, 30s, 60s, 120s). 5-ethynyl-2'-deoxyuridine (EdU) proliferation assay was used to determine the cell viability. Analysis of cell migration and invasion was performed by wound-healing assay, transwell assay and immunofluorescent staining for f-actin, respectively. Flow cytometry technique was used for analysis of cell cycle and determination of reactive oxygen species (ROS). Hoechst staining was used for analysis of cell apoptosis. Protein expression was analyzed by Western blot analysis. Results: Molecular and cellular level mechanisms have revealed that CAP blocks RA-FLS in the G2/M phase by increasing intracellular reactive oxygen species (ROS), leading to increased apoptosis and significantly reduced migration and invasion ability of RA-FLS. Conclusion: Overall, CAP has significant anti proliferative, migratory, and invasive effects on RA-FLS. This study reveals a new targeted treatment strategy for RA.

Mutations of epigenetic modifier genes predict poor outcome in adult acute lymphoblastic leukemia.

Epigenetic modifier (EM) genes play important roles in the occurrence and progression of acute lymphoblastic leukemia (ALL). However, the prognostic significance of EM mutations in ALL has not yet been thoroughly investigated. This retrospective study included 205 adult patients with ALL engaged in a pediatric-type regimen. Based on targeted next-generation sequencing, they were divided into EM mutation group (EM-mut, n = 75) and EM wild-type group (EM-wt, n = 130). The EM-mut group showed a higher positive rate of minimal residual disease (MRD) on treatment day24 and before consolidation therapy (P = 0.026, 0.020). Multivariate Cox regression analysis showed that EM-mut was an independent adverse factor for overall survival (OS) and event-free survival (EFS) (HR = 2.123, 1.742; P = 0.009, 0.007). Survival analysis revealed that the OS and EFS rates were significantly lower in the EM-mut group than in the EM-wt group (3-year OS rate, 45.8% vs. 65.0%, P = 0.0041; 3-year EFS rate, 36.7% vs. 53.2%, P = 0.011). In conclusion, EM was frequently mutated in adult ALL and was characterized by poor response to induction therapy and inferior clinical outcomes.

Cold atmospheric plasma stabilizes mismatch repair for effective, uniform treatment of diverse colorectal cancer cell types.

Mismatch Repair (MMR) mechanisms play a pivotal role in rectifying DNA replication errors and maintaining the stability of DNA microsatellite structure. Colorectal cancer (CRC) can be characterized into microsatellite stability (MSS) and microsatellite instability (MSI) subtypes based on the functionality of MMR. MSI CRC notably exhibits enhanced chemotherapy resistance, attributable to diminished MMR-related protein expression. Cold atmospheric plasma (CAP) has emerged as a promising treatment modality, demonstrating efficacy in inducing apoptosis in various cancer cells. However, the therapeutic impact of CAP on MSI colorectal cancer, and the underlying mechanisms remain elusive. In this study, we investigated the effects of CAP on MSI (MC38, HCT116, and LOVO) and MSS (CT26 and HT29) CRC cell lines. We are probing into the products of CAP treatment. Our findings indicate that CAP treatment induces comparable effects on apoptosis, reactive oxygen species (ROS), and reactive nitrogen species (RNS), as well as the expression of apoptosis-related proteins in both MSI and MSS cells. Mechanistically, CAP treatment led to an elevation in the expression of mismatch repair proteins (MLH1 and MSH2), particularly in MSI cells, which notably have been proven to facilitate the activation of apoptosis-related proteins. Collectively, our study reveals that CAP enhances apoptotic signaling and induces apoptosis in MSI colorectal cancer cells by upregulating the expression of MMR-related proteins, thereby reinforcing MMR stabilization.

Design of a 140 GHz waveguide notch filter for millimeter-wave receiver module protection in fusion plasma diagnostics.

A carefully designed waveguide-based millimeter-wave notch filter, operating at 140 GHz, safeguards plasma diagnostic instruments from gyrotron leakage. Utilizing cylindrical cavity resonators with aperture coupling, the filter efficiently resonates 140 GHz wave-power into the TE11p mode, optimizing various geometrical parameters for practical fabrication and high-yield production. Thorough thermal analysis ensures its ability to handle power. The filter achieves outstanding performance with over 90 dB rejection at 140 GHz while providing low insertion loss over the passband (110-138 GHz), which is ideally suited for system-on-chip approach F-band diagnostic system applications.

Cold plasma turns mixed-dye-contaminated wastewater bio-safe.

The excessive and uncontrollable discharge of diverse organic pollutants into the environment has emerged as a significant concern, presenting a substantial risk to human health. Among the advanced oxidation processes used for the purification of wastewater, cold plasma technology is superior in fast and effective decontamination but often fails facing mixed pollutants. To address these issues, here we develop the new conceptual approach, plasma process, and proprietary reactor that ensure, for the first time, that the efficiency of treatment (114.7%) of two mixed organic dyes, methylene blue (MB) and methyl orange (MO), is higher than when the two dyes are treated separately. We further reveal the underlying mechanisms for the energy-efficient complete degradation of the mixed dyes. The contribution of plasma-induced ROS and the distinct degradation characteristics and mechanism of pollutants in mixed treatment are discussed. The electron transfer pathway revealed for the first time suggest that the mixed pollutants reduce the overall redox potentials and facilitate electron transfer during the plasma treatment, promoting synergistic degradation effects. The integrated frameworks including both direct and indirect mechanisms provide new insights into the high-efficiency mixed-contaminant treatment. The degradation products for mixed degradation are revealed based on the identification of intermediate species. The plasma-treated water is proven safe for living creatures in waterways and sustainable fishery applications, using in vivo zebrafish model bio-toxicity assay. Overall, these findings offer a feasible approach and new insights into the mechanisms for the development of highly-effective, energy-efficient technologies for wastewater treatment and reuse in agriculture, industry, and potentially in urban water networks.

Cotton Fibers with a Lactic Acid-Like Surface for Re-establishment of Protective Lactobacillus Microbiota by Selectively Inhibiting Vaginal Pathogens.

Failure to reconstruct the Lactobacillus microbiota is the major reason for the recurrence of vaginal infection. However, most empiric therapies focus on the efficacy of pathogen elimination but do not sufficiently consider the viability of Lactobacillus. Herein, cotton fibers with a lactic acid-like surface (LC) are fabricated by NaIO4 oxidation and L-isoserine grafting. The lactic acid analog chain ends and imine structure of LC can penetrate cell walls to cause protein cleavage in Escherichia coli and Candida albicans and inhibit vaginal pathogens. Meanwhile, the viability of Lactobacillus acidophilus is unaffected by the LC treatment, thus revealing a selective way to suppress pathogens as well as provide a positive route to re-establish protective microbiota in the vaginal tract. Moreover, LC has excellent properties such as good biosafety, antiadhesion, water absorption, and weight retention. The strategy proposed here not only is practical, but also provides insights into the treatment of vaginal infections.

In vitro evaluation of BMSCs early proliferation on minocycline-loaded electrospun nanofibers membrane.

BACKGROUND: Electrospun nanofibers could simulate the natural extracellular matrix (ECM) of the host bone, while minocycline (MINO) is a broad-spectrum tetracycline antibiotic which has been found to have multiple non-antibiotics biological effects that promotes osteogenesis in vitro and in vivo. OBJECTIVE: The present study aims at constructing a polylactic acid (PLA) electrospun nanofiber membrane loaded with MINO to enhance Bone marrow mesenchymal stem cells (BMSCs) adhesion and proliferation for early clinical treatment. METHODS: The MINO-PLA membrane were characterized by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR) and in vitro drug release study. The antibacterial ability was also investigated. In addition, in vitro cellular proliferation experiment was performed to verify whether the PLA electrospun nanofibers membrane loaded with MINO enhance BMSCs adhesion and proliferation. RESULTS: Analyzing the drug release and cell growth results, it was found that only the effective concentration of MINO-PLA could help the growth of BMSCs in the short term. This is related to the drug release rate of MINO-PLA and the initial concentration of MINO. CONCLUSION: This study shows that by controlling the concentration and release rate of MINO with electrospinning PLA, BMSCs could proliferate on it, and a new bone repair material had been made in this study.

PASS-ALL study of paediatric-inspired versus adult chemotherapy regimens on survival of high-risk Philadelphia-negative B-cell acute lymphoblastic leukaemia with allogeneic haematopoietic stem cell transplantation.

This PASS-ALL study was designed to explore the effect of paediatric-inspired versus adult chemotherapy regimens on survival of adolescents and young adults (AYA) with high-risk Philadelphia chromosome-negative B-cell acute lymphoblastic leukaemia (HR PH-ve B-cell ALL) eligible for allogeneic haematopoietic stem cell transplantation (allo-HSCT). The PASS-ALL study is a multicentre, observational cohort study, and 143 patients with HR B-cell PH-ve ALL were enrolled from five centres-77 patients allocated in the paediatric-inspired cohort and 66 in the adult cohort with comparable baseline characteristics. Of the 143 patients, 128 cases underwent allo-HSCT. Three-year leukaemia-free survival (LFS) in the paediatric-inspired cohort was 72.2% (95% CI 60.8%-83.6%) compared with 44.6% (95% CI 31.9%-57.3%; p = 0.001). Furthermore, time-to-positive minimal residual disease (TTP-MRD) post-HSCT was marked different, 3-year cumulative incidence of relapse was 25.9% (95% CI 15.8%-37.2%) in paediatric cohort and 45.4% (95% CI 40.0%-57.9%) in adult cohort (p = 0.026). Finally, the 3-year OS rate was 75.3% (95% CI 64.9%-85.7%) for the paediatric-inspired cohort and 64.1% (95% CI 51.8%-76.4%) for the adult cohort (p = 0.074). On a multivariate analysis, paediatric-inspired regimen is a predictive factor for LFS (HR = 2.540, 95% CI 1.327-4.862, p = 0.005). Collectively, our data suggest that paediatric-inspired chemotherapy pre-HSCT results in deeper and durable MRD response reduces relapse post-HSCT and improves survival in HR B-cell PH-ve ALL patients with allo-HSCT.

Diffusion-driven fabrication of calcium and phosphorous-doped zinc oxide heterostructures on titanium to achieve dual functions of osteogenesis and preventing bacterial infections.

Conventional Ti-based implants are vulnerable to postsurgical infection and improving the antibacterial efficiency without compromising the osteogenic ability is one of the key issues in bone implant design. Although zinc oxide (ZnO) nanorods grown on Ti substrates hydrothermally can improve the antibacterial properties, but cannot meet the stringent requirements of bone implants, as rapid degradation of ZnO and uncontrolled leaching of Zn2+ are detrimental to peri-implant cells and tissues. To solve these problems, a lattice-damage-free method is adopted to modify the ZnO nanorods with thin calcium phosphate (CaP) shells. The Ca and P ions from the CaP shells diffuse thermally into the ZnO lattice to prevent the ZnO nanorods from rapid degradation and ensure the sustained release of Zn2+ ions as well. Furthermore, the designed heterostructural nanorods not only induce the osteogenic performances of MC3T3-E1 cells but also exhibit excellent antibacterial ability against S. aureus and E. coli bacteria via physical penetration. In vivo studies also reveal that hybrid Ti-ZnO@CaP5 can not only eradicates bacteria in contact, but also provides sufficient biocompatibility without causing excessive inflammation response. Our study provides insights into the design of multifunctional biomaterials for bone implants. STATEMENT OF SIGNIFICANCE: • A lattice-damage-free method is adopted to modify the ZnO nanorods with thin calcium phosphate (CaP) shells. • The dynamic process of Ca and P diffusion into the ZnO lattice is analyzed by experimental verification and theoretical calculation. • The degradation rate of ZnO nanorods is significantly decreased after CaP deposition. • The ZnO nanorods after CaP deposition can not only sterilize bacteria in contact via physical penetration, but also provide sufficient biocompatibility and osteogenic capability without causing excessive inflammation response..

Inverse design of an ultra-compact and large-bandwidth bent subwavelength grating wavelength demultiplexer.

Inverse design has attracted significant attention as a method to improve device performance and compactness. In this research, we employed a combination of forward design and the inverse algorithm using particle swarm optimization (PSO) to design a bent ultra-compact 1310/1550 nm broadband wavelength demultiplexer assisted by a subwavelength grating (SWG). Through the phase matching at 1550 nm and the phase mismatch at 1310 nm, we rapidly designed the width parameters of SWG in the forward direction. Then the PSO algorithm was used to optimize the SWG parameters in a certain range to achieve the best performance. Additionally, we introduced a new bent dimension significantly reducing the device length while maintaining low insertion loss (IL) and high extinction ratios (ERs). It has been verified that the length of the device is only 7.8 µm, and it provides a high ER of 24 dB at 1310 nm and 27 dB at 1550 nm. The transmitted spectrum shows that the IL values at both wavelengths are below 0.1 dB. Meanwhile, the 1 dB bandwidth exceeds 150 nm, effectively covering the entire O-band and C-band. This approach has been proven successful in enhancing performance and significantly reducing the device footprint.

Bactericidal effect of low-temperature atmospheric plasma against the Shigella flexneri.

BACKGROUND: Shigella flexneri (S. flexneri) is a common intestinal pathogenic bacteria that mainly causes bacillary dysentery, especially in low socioeconomic countries. This study aimed to apply cold atmospheric plasma (CAP) on S. flexneri directly to achieve rapid, efficient and environmentally friendly sterilization. METHODS: The operating parameters of the equipment were determined by plasma diagnostics. The plate count and transmission electron microscope were employed to calculate bacterial mortality rates and observe the morphological damage of bacterial cells. Measurement of intracellular reactive oxygen species (ROS) and superoxide anions were detected by 2,7-dichlorodihydrofluorescein (DCFH) and Dihydroethidium fluorescence probes, respectively. The fluorescence intensity (a. u.) reflects the relative contents. Additionally, the experiment about the single effect of temperature, ultraviolet (UV), and ROS on bacteria was conducted. RESULTS: The peak discharge voltage and current during plasma operation were 3.92kV and 66mA. After discharge, the bacterial mortality rate of 10, 20, 30 and 40 s of plasma treatment was 60.71%, 74.02%, 88.11% and 98.76%, respectively. It was shown that the intracellular ROS content was proportional to the plasma treatment time and ROS was the major contributor to bacterial death. CONCLUSION: In summary, our results illustrated that the plasma treatment could inactivate S. flexneri efficiently, and the ROS produced by plasma is the leading cause of bacterial mortality. This highly efficient sterilization method renders plasma a highly promising solution for hospitals, clinics, and daily life.

Epigenetic dysregulation of eukaryotic initiation factor 3 subunit E (eIF3E) by lysine methyltransferase REIIBP confers a pro-inflammatory phenotype in t(4;14) myeloma.

REIIBP is a lysine methyltransferase aberrantly expressed through alternative promoter usage of NSD2 locus in t(4;14)-translocated multiple myeloma (MM). Clinically, t(4;14) translocation is an adverse prognostic factor found in approximately 15% of MM patients. The contribution of REIIBP relative to other NSD2 isoforms as a dependency gene in t(4;14)-translocated MM remains to be evaluated. Here, we demonstrated that despite homology with NSD2, REIIBP displayed distinct substrate specificity by preferentially catalyzing H3K4me3 and H3K27me3, with little activity on H3K36me2. Furthermore, REIIBP was regulated through microRNAs by EZH2 in a Dicer-dependent manner, exemplifying a role of REIIBP in SET-mediated H3K27me3. ChIP-sequencing revealed chromatin remodeling characterized by changes in genome-wide and loci-specific occupancy of these opposing histone marks, allowing a bidirectional regulation of its target genes. Transcriptomics indicated that REIIBP induced a pro-inflammatory gene signature through upregulation of TLR7, which in turn led to B-cell receptor (BCR)-independent activation of BTK and driving NFĸB-mediated production of cytokines such as IL-6. Activation of this pathway is targetable using Ibrutinib and partially mitigated bortezomib resistance in an REIIBP xenograft model. Mechanistically, REIIBP upregulated TLR7 through eIF3E, and this relied on eIF3E RNA-binding function instead of its canonical protein synthesis activity, as demonstrated by direct binding to the 3'UTR of TLR7 mRNA. Altogether, we provided a rationale that coexistence of different NSD2 isoforms induced diversified oncogenic programs that should be considered in the strategies for t(4;14)-targeted therapy.

Bio-orthogonal engineered peptide: A multi-functional strategy for the gene therapy of osteoporotic bone loss.

Osteoporosis is a degenerative disease affecting millions of elderly people globally and increases the risk of bone fractures due to the reduced bone density. Drugs are normally prescribed to treat osteoporosis, especially after surgical treatment of osteoporotic fractures. However, many anti-osteoporotic drugs produce deleterious side effects. The recent development of gene therapy utilizing oligonucleotides (ONs) has spurred the development of new therapies for osteoporosis. Nevertheless, most ONs lack the capability of cell penetration and lysosome escape and hence, intracellular delivery of ON remains a challenge. Herein, a novel strategy is demonstrated to efficiently deliver ON to cells by combining ON with the cell-penetrating peptide (CPP) via the bio-orthogonal click reaction. Several dopamine (DOPA) groups are also introduced into the fabricated peptide to scavenge intracellular reactive oxygen species (ROS). Owing to favorable properties such as good cytocompatibility, cell penetration, lysosome escape, ROS scavenging, and osteoclastogenesis suppression, the hybrid CPP-DOPA-ON peptide improves the osteoporotic conditions significantly in vivo even when bone implants are involved. This strategy has great potential in the treatment of osteoporosis and potentially broadens the scope of gene therapy.

All-optical spiking neural network and optical spike-time-dependent plasticity based on the self-pulsing effect within a micro-ring resonator.

In this paper, we proposed an all-optical version of photonic spiking neurons and spike-time-dependent plasticity (STDP) based on the nonlinear optical effects within a micro-ring resonator. In this system, the self-pulsing effect was exploited to implement threshold control, and the equivalent pulse energy required for spiking, calculated by multiplying the input pulse power amplitude with its duration, was about 14.1 pJ. The positive performance of the neurons in the excitability and cascadability tests validated the feasibility of this scheme. Furthermore, two simulations were performed to demonstrate that such an all-optical spiking neural network incorporated with STDP could run stably on a stochastic topology. The essence of such an all-optical spiking neural network is a nonlinear spiking dynamical system that combines the advantages of photonics and spiking neural networks (SNNs), promising access to the high speed and lower consumption inherent to optical systems.

Cold atmospheric pressure plasma: A potential physical therapy for rheumatoid arthritis hyperplastic synovium.

The most significant pathological change in rheumatoid arthritis (RA) is synovial hyperplasia within the joint. The production of a series of degrading enzymes and oxidative stress caused by synovial hyperplasia lead to severe bone and cartilage damage in rheumatoid joints. The core effector cell in hyperplastic synovium is fibroblast-like synovium cells, which can invade cartilage, cause inflammation, destroy joints, and show tumor-like anti-apoptosis characteristics. This study focused on the effect of cold atmospheric pressure plasma on proliferative synovium, and the results showed that no synovial hyperplasia, angiogenesis, or inflammatory infiltration was observed after cold atmospheric pressure plasma (CAP) treatment. The molecular and cellular mechanisms also reveal the spontaneous reactive oxygen species (ROS) cascade inducing apoptosis in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) cells. This study proposes a potential physical therapy method for treating proliferative synovium and also provides ideas for the application of CAP in other types of tumor diseases.

High-Sensitivity Graphene MOEMS Resonant Pressure Sensor.

Nanomechanical resonators made from suspended graphene exhibit high sensitivity toward pressure variations. Nevertheless, these devices exhibit significant energy loss in nonvacuum environments due to air damping, as well as inevitably weak gas leakage within the reference cavity because of the slight permeation of graphene. We present a new type of graphene resonant pressure sensor utilizing micro-opto-electro-mechanical systems technology, which features a multilayer graphene membrane that is sealed in vacuum and adhered to pressure-sensitive silicon film with grooves. This approach innovatively employs an indirectly sensitive method, exhibiting 60 times smaller energy loss in atmosphere, and solving the long-standing issue of gas permeation between the substrate and graphene. Notably, the proposed sensor exhibits a high pressure sensitivity of 1.7 Hz/Pa, which is 5 times higher than the sensitivity of the silicon counterparts. Also, the all-optical encapsulating cavity structure contributes a high signal-to-noise ratio of 6.9 × 10-5 Pa-1 and a low temperature drift (0.014%/◦C). The proposed method offers a promising solution for long-term stability and energy loss suppression of pressure sensors using two-dimensional materials as the sensitive membrane.

The effect of inhibition of receptor tyrosine kinase AXL on DNA damage response in ovarian cancer.

AXL is a receptor tyrosine kinase that is often overexpressed in cancers. It contributes to pathophysiology in cancer progression and therapeutic resistance, making it an emerging therapeutic target. The first-in-class AXL inhibitor bemcentinib (R428/BGB324) has been granted fast track designation by the U.S. Food and Drug Administration (FDA) in STK11-mutated advanced metastatic non-small cell lung cancer and was also reported to show selective sensitivity towards ovarian cancers (OC) with a Mesenchymal molecular subtype. In this study, we further explored AXL's role in mediating DNA damage responses by using OC as a disease model. AXL inhibition using R428 resulted in the increase of DNA damage with the concurrent upregulation of DNA damage response signalling molecules. Furthermore, AXL inhibition rendered cells more sensitive to the inhibition of ATR, a crucial mediator for replication stress. Combinatory use of AXL and ATR inhibitors showed additive effects in OC. Through SILAC co-immunoprecipitation mass spectrometry, we identified a novel binding partner of AXL, SAM68, whose loss in OC cells harboured phenotypes in DNA damage responses similar to AXL inhibition. In addition, AXL- and SAM68-deficiency or R428 treatment induced elevated levels of cholesterol and upregulated genes in the cholesterol biosynthesis pathway. There might be a protective role of cholesterol in shielding cancer cells against DNA damage induced by AXL inhibition or SMA68 deficiency.

A Meaningful Attempt: Applying Dielectric Barrier Discharge Plasma to Induce Polarization of Macrophages.

Macrophage polarization plays an important role in many macrophage-related diseases. This study was designed to preliminarily explore the effects of dielectric barrier discharge (DBD) plasma on the polarization direction and cell activity of macrophages with different phenotypes (ie, M0, M1, and M2). The M1 macrophage marker inducible nitric oxide synthase (iNOS) and M2 macrophage marker cluster of differentiation 206 (CD206) were detected by western blot (WB). The effects of DBD plasma on macrophage viability were analyzed by using a cell counting kit-8 detection kit. M0, M1, and M2 macrophages exhibited a decrease in iNOS expression and an increase in CD206 expression after the DBD plasma intervention. Additionally, the decrease in macrophage viability remained non-significant after initiating the intervention. DBD plasma can promote the transformation of M0 and M1 macrophages to M2 macrophages, and can further enhance the expression of the M2 macrophage phenotype marker CD206. Our study not only demonstrates the potential therapeutic value of DBD plasma for macrophage-related diseases, but it also provides a new direction for research to improve the treatment of macrophage-related diseases. © 2023 Bioelectromagnetics Society.

Non-thermal plasma-treated melatonin inhibits the biological activity of HCC cells by increasing intracellular ROS levels and reducing RRM2 expression.

Non-thermal plasma (NTP) is thought to have a cytotoxic effect on tumor cells. Although its application in cancer therapy has shown considerable promise, the current understanding of its mechanism of action and cellular responses remains incomplete. Furthermore, the use of melatonin (MEL) as an adjuvant anticancer drug remains unexplored. In this study, we found that NTP assists MEL in promoting apoptosis, delaying cell cycle progression, and inhibiting cell invasion and migration in hepatocellular carcinoma (HCC) cells. This mechanism may be associated with the regulation of intracellular reactive oxygen species levels and ribonucleotide reductase regulatory subunit M2 expression. Our findings confirm the pharmacological role of MEL and the adjuvant value of NTP, emphasizing their potential in combination therapy for HCC. Our study may have important implications for the development of new approaches for HCC treatment.