A dual-channel fluorescent probe targeting lysosomes for differential detection of Cys/Hcy and GSH: Applications in food, pharmaceutical analysis and bioimaging.

Thiols function as antioxidants in food, prolonging shelf life and enhancing flavor. Moreover, thiols are vital biomolecules involved in enzyme activity, cellular signal transduction, and protein folding among critical biological processes. In this paper, the fluorescent probe PYL-NBD was designed and synthesized, which utilized the fluorescent molecule pyrazoline, the lysosome-targeted morpholine moiety, and the sensing moiety NBD. Probe PYL-NBD was tailored for the recognition of biothiols through single-wavelength excitation, yielding distinct fluorescence emission signals: blue for Cys, Hcy, and GSH; green for Cys, Hcy. Probe PYL-NBD exhibited rapid reaction kinetics (<10 min), distinct fluorescence response signals, and low detection limits (15.7 nM for Cys, 14.4 nM for Hcy, and 12.6 nM for GSH). Probe PYL-NBD enabled quantitative determination of Cys content in food samples and L-cysteine capsules. Furthermore, probe PYL-NBD had been successfully applied for confocal imaging with dual-channel detection of biothiols in various biological specimens, including HeLa cells, zebrafish, tumor sections, and Arabidopsis thaliana.

Transjugular intrahepatic portosystemic shunt for esophagogastric variceal bleeding in patients with hepatocellular carcinoma and portal vein tumor thrombus.

BACKGROUND: Whether hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT) and acute esophagogastric variceal bleeding (EGVB) can improve the success rate of endoscopic hemostasis and overall survival (OS) from transjugular intrahepatic portosystemic shunt (TIPS) remains controversial. AIM: To compare the clinical outcomes between TIPS and standard treatment for such HCC patients. METHODS: This monocenter, retrospective cohort study included patients diagnosed as HCC with PVTT and upper gastrointestinal bleeding. Patients were grouped by the treatment (TIPS or standard conservative treatment). The success rate of endoscopic hemostasis, OS, rebleeding rates, and main causes of death were analyzed. RESULTS: Between July 2015 and September 2021, a total of 77 patients (29 with TIPS and 48 with standard treatment) were included. The success rate of endoscopic hemostasis was 96.6% in the TIPS group and 95.8% in the standard treatment group. All the 29 patients in TIPS group successful underwent TIPS procedure and had a better OS compared with standard treatment within the first 160 days after treatment (68 days vs 43 days, P = 0.022), but shorter OS after 160 days (298 days vs 472 days, P = 0.022). Cheng's Classification of PVTT, total bilirubin and Child-Pugh class were independently negative associated with OS (all P < 0.05). The main causes of death were liver failure or hepatic encephalopathy (75.9%) in the TIPS group and rebleeding (68.8%) in the standard treatment. CONCLUSION: TIPS could reduce the risk of early death due to rebleeding and prolong short-term survival in HCC patients with PVTT and acute EGVB, which deserves further investigation.

Enhanced nitrogen removal for low C/N wastewater via preventing futile carbon oxidation and augmenting anammox.

Efficient carbon use is crucial for biological nitrogen removal. Traditional aerobic processes can waste carbon sources, exacerbating carbon deficiency. This study explores an anaerobic/oxic/anoxic system with sludge double recirculation to improve nitrogen removal in low C/N wastewater. This system integrated aerobic nitrification after the carbon intracellular storage, separating carbon and nitrogen by denitrifying glycogen-accumulating organisms (DGAOs) with endogenous partial denitrification and Anammox within the anoxic units. A significant efficiency of 91.02±7.01% chemical oxygen demand (COD) was converted into intracellular carbon in anaerobic units, significantly reducing carbon futile oxidation in the aerobic units by effectively separating COD from ammonia. Intracellular storage of carbon sources and microbial adaptation to carbon scarcity prevent futile oxidation of COD in the aerobic units even with short-term high dissolved oxygen (DO), thereby enhancing nitrogen removal under anoxic conditions with sufficient intracellular carbon source. The microbial analysis identified Candidatus Brocadia as the dominant anammox bacteria, in combination with the activity of DGAOs and other related microbial communities, accounting for 37.0% of the TN removal. Consequently, the system demonstrated remarkable nitrogen removal efficiencies, achieving 81.3±3.3% for total nitrogen (TN) and 98.5±0.9% for ammonia nitrogen while maintaining an effluent COD concentration of 17.2±9.1 mg/L, treating the low C/N of 4.18 in the influent wastewater. The findings in this study provide a sustainable and energy-saving technique for conventional WWTPs to meet strict discharge standards by avoiding futile oxidation of COD and encouraging anammox contributions.

TiO2 nanotube enhance osteogenesis through Kindlin-2/Integrin ß1/YAP pathway-mediated mechanotransduction.

Titanium has been widely employed in the fields of orthopaedics and dentistry, attributed to its superior mechanical and biological properties. The mechanical stimulation induced by the titanium dioxide (TiO2) nanotubes (TNTs) morphology resulting from surface modification has been demonstrated to enhance the osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs). Kindlin-2, a pivotal focal adhesion protein, is involved in mechanical signaling processes through the regulation of stress fibril filament assembly. Additional research is needed to clarify the involvement of Kindlin-2 in the mechanism of TNTs-induced osteogenic differentiation. This study systematically investigated the impact of Kindlin-2 on TNTs-induced osteogenesis and mechanotransduction. TiO2 nanotubes with diameters of approximately 30 nm (TNT-30) and 100 nm (TNT-100) were fabricated and characterized using anodic oxidation. The results showed that TNT-100 significantly increased the expression of Kindlin-2 and enhanced osteogenic differentiation compared to polished titanium (PT) and TNT-30. Additionally, Kindlin-2 promotes cytoskeleton assembly by regulating the integrin ß1/FAK/RhoA signaling pathway, impacting osteogenic gene expression and BMSC differentiation in a Yes-Associated Protein (YAP)-dependent manner. Therefore, these findings contribute to a more comprehensive understanding of the fate of BMSCs on TNTs morphologies and provide a novel theoretical foundation for the development of advanced bone repair biomaterials.

Suicidality, perceived chronic stress, and stress-induced cortisol changes of individuals with schizophrenia.

The associations of suicidality with stress are poorly studied in schizophrenia. The study aimed to determine whether suicidality was correlated with perceived chronic stress and the cortisol fluctuations under stress tasks in schizophrenia. High suicidality was defined as a lifetime history of suicide attempts or suicidal ideation in the past 2 weeks. Individuals with schizophrenia and high suicidality (SZ-HS, n = 59), with low suicidality (SZ-LS, n = 207), and healthy controls (HC, n = 196) finished the Perceived Stress Scale. Then, they participated in an experiment that induced stress using the Paced Auditory Serial Addition Task and the Mirror Tracing Persistence Task. Negative affect was measured at baseline and after finishing each task. The salivary cortisol was collected before-, after 20 min, and after 40 min of the tasks. SZ-HS had elevated perceived stress than SZ-LS and HC. Mixed effect models showed that stress tasks induced cortisol changes in all groups; cortisol of SZ-LS was reduced more than HC, but SZ-HS and SZ-LS did not differ in cortisol fluctuations. SZ-HS and SZ-LS experienced similar negative affect changes during tasks and the difference in withdrawal rates was nonsignificant. SZ-HS had an increased error rate than SZ-LS. In conclusion, suicidality was correlated with high-level perceived stress but did not result in differences in cortisol reactivities under stress tasks. It suggests the inconsistency between appraisal of stress and biological stress system disturbance among SZ-HS compared to SZ-LS.

Increased Risk of Short-Term Infection After Liver Transplantation Combined With Splenectomy: Clinical Retrospective Study.

AIM: The safety of liver transplantation and simultaneous splenectomy (LTSP) is still controversial. This study aimed to compare postoperative outcomes and infection in liver transplant recipients with and without simultaneous splenectomy. METHODS: Clinical data of patients who underwent liver transplantation (LT) from May 2015 to March 2023 in the First Affiliated Hospital of Anhui Medical University were retrospectively analyzed. The main parameters measured were culture results, infection incidence, pathogens, postoperative complications, and overall survival rates. RESULTS: Of 149 patients, 35 who underwent LTSP were assigned to the LTSP group, and the remaining 114 were assigned to the LT group. The postoperative infection incidence in the LTSP group was significantly higher than in the LT group within 1 month after transplantation. The two groups had no significant differences in pathogens details and overall survival rate. SP, postoperative days (POD) 3 Neutrophil to lymphocyte ratio (NLR), POD 7 NLR, and POD 7 Hemoglobin (HGB) were independent risk factors for postoperative infection in multivariate analysis. CONCLUSION: LTSP increases the risk of short-term postoperative infections, and postoperative NLR can be used as a marker of infection.

Fabrication and in vitro biocompatibility of hierarchical cellulose acetate/polyvinylpyrrolidone@titania nanowire hollow microfibers.

In this study, hierarchical cellulose acetate/polyvinylpyrrolidone hollow microfibers (CA/PVP HMFs) were first prepared via a dip coating method using a steel wire as tubular template and then supported a sol-gel deposition of titania nanoparticles (NPs) to derive CA/PVP@titania NP HMFs. After hydrothermally treated in NaOH solution, CA/PVP@titania NP HMFs were transformed to CA/PVP@titania nanowire (NW) HMFs. SEM observation showed that CA/PVP@titania NW HMFs had a hollow structure with diameters of 450-600 µm and exhibited a hierarchical and nanofibrous structure. Their surfaces were constructed by numerous titania NWs with diameters of 10-30 nm and lengths of 1-5 µm. The incorporation of PVP not only caused a significant change in surface wettability from hydrophobic CA HMFs to hydrophilic CA/PVP HMFs, but also promoted the sol-gel deposition of titania NPs on CA/PVP HMFs. CA/PVP@titania NW HMFs exhibited the highest hydrophilicity with water contact angle of 32° and the largest specific surface area of 86.1 m2/g. In vitro biocompatible evaluation indicated that CA/PVP@titania NW HMFs exhibited much higher cell adhesion and proliferation than CA/PVP@titania NP HMFs and CA/PVP HMFs within 7 days due to the presence of nanofibrous surface architecture. Thus, the present CA/PVP titania NW HMFs have potential as biocompatible cell supporting matrices.

[Pollution Assessment and Source Apportionment of Heavy Metals in the Surrounding Soil of Typical Mining Areas in Tongling, Anhui Province].

To study the level of heavy metal pollution and ecological risks in the soil around typical mining areas in Tongling, a total of 150 soil samples were collected from the study area. The content characteristics of 10 elements, namely, As, Cd, Cr, Cu, Hg, Mn, Ni, Pb, Fe, and Zn, in the soils were analyzed. Methods including enrichment factor, the geo-accumulation index, single-factor pollution index, Nemero comprehensive pollution index, and potential ecological risk index were used to evaluate the pollution status of heavy metals in the soil of the study area. The pollution sources of heavy metals in the soil were also analyzed using correlation analysis, cluster analysis, and principal component analysis. The results showed that except for Cr and Fe, the average contents of the other eight heavy metal elements were higher than the soil background values in the study area. Pb, Zn, As, Cu, and Cd had a high degree of variation and were significantly affected by external interference. The spatial distribution showed that both Cr and Ni showed a decreasing trend from the edge to the central region, whereas the other eight heavy metals showed a decreasing trend from the central region to the surrounding areas. The pollution level of Cd and Cu in the soil of the research area was relatively severe. The overall ecological risk was at a medium to low level. Cd and Hg were the main contributing factors. As, Cd, Cu, Fe, Mn, Pb, and Zn mainly came from agricultural, industrial, and transportation sources, whereas Cr and Ni were mainly from natural sources. However, the sources of Hg were relatively complex. The research results can provide a scientific basis for the prevention and control of soil heavy metal pollution in metal mining areas, as well as the remediation of mine pollution.

Spatial Structure of Electron Interactions in High-entropy Oxide Nanoparticles for Active Electrocatalysis of Carbon Dioxide Reduction.

High-entropy oxides (HEOs) exhibit distinctive catalytic properties owing to their diverse elemental compositions, garnering considerable attention across various applications. However, the preparation of HEO nanoparticles with different spatial structures remains challenging due to their inherent structural instability. Herein, ultrasmall high-entropy oxide nanoparticles (less than 5 nm) with different spatial structures are synthesized on carbon supports via the rapid thermal shock treatment. The low-symmetry HEO, BiSbInCdSn-O4, demonstrates exceptional performance for electrocatalytic carbon dioxide reaction (eCO2RR), including a lower overpotential, high Faraday efficiency across a wide electrochemical range (-0.3 to -1.6 V), and sustained stability for over100 h. In the membrane electrode assembly electrolyzer, BiSbInCdSn-O4 achieves a current density of 350 mA cm-2 while maintaining good stability for 24 h. Both experimental observations and theoretical calculations reveal that the electron donor-acceptor interactions between bismuth and indium sites in BiSbInCdSn-O4 enable the electron delocalization to facilitate the efficient adsorption of CO2 and hydrogenation reactions. Thus, the energy barrier of the rate-determining step is reduced to enhance the electrocatalytic activity and stability. This study elucidates that the spatial structure of metal sites in HEOs is able to regulate CO2 adsorption status for eCO2RR, paving the way for the rational design of efficient HEO catalysts.

Receptor-interacting protein kinase 1 confers autophagic promotion of gasdermin E-mediated pyroptosis in aristolochic acid-induced acute kidney injury.

Aristolochic acid (AA) exposure is a severe public health concern worldwide. AAs damage the kidney with an inevitable acute phase that is similar to acute kidney injury (AKI). Gasdermin E (GSDME) is abundant in the kidney; thus; it-mediated pyroptosis might be essential in connecting cell death and inflammation and promoting AAs-AKI. However, the role and exact mechanism of pyroptosis in AAs-AKI have not been investigated. In this study, aristolochic acid I (AAI) was used to establish AKI models. The expression and translocation results showed GSDME-mediated pyroptosis in AAI-AKI. Knocking down GSDME attenuated AAI-induced cell death and transcription of proinflammatory cytokines. Mechanistic research inhibiting caspase (casp) 3, casp 8, and casp 9 with specific chemical antagonists demonstrated that GSDME was activated by cleaved casp 3. Furthermore, the kinase activity of upstream receptor-interacting protein kinase 1 (RIPK1) was significantly elevated, and inhibiting RIPK1 with specific inhibitors markedly improved AAI-induced cell damage. In addition, the level of autophagy was obviously increased. Pretreatment with a specific autophagic inhibitor (3-methyladenine) or knockdown of autophagic genes (Atg5 or Atg7) evidently reduced the activity of RIPK1 and downstream apoptosis and pyroptosis, thus attenuating AA-induced cell injury, which suggested that RIPK1 was a novel link conferring autophagic promotion of pyroptosis. These findings reveal GSDME-mediated pyroptosis for the first time in AAI-induced AKI, propose its novel role in the transcription of cytokines, and demonstrate that autophagy promotes pyroptosis via the RIPK1-dependent apoptotic pathway. This study promotes the understanding of the toxic effects and exact mechanisms of AAs. This will contribute to evaluating the environmental risk of AA exposure and might provide potential therapeutic targets for AA-AKI.

A lysosome-targeted fluorescent probe for thiol detection in drug analysis and multiple biological systems.

Biothiols, characterized by their unique sulfhydryl (-SH) groups, possess excellent antioxidant properties, effectively neutralizing the damage to cellular structures caused by reactive oxygen species (ROS) in living organisms. Additionally, lysosomes play a crucial role in decomposing damaged biomolecules through the action of their internal enzymes, regulating the cellular redox state, and mitigating oxidative stress. To facilitate rapid monitoring of intracellular biothiols, particularly within lysosomes, we constructed a lysosome-targeted biothiol fluorescent probe, PHL-DNP, in this study. PHL-DNP exhibited excellent photophysical properties in an aqueous test system, including strong fluorescence enhancement response, excellent selectivity, and low detection limits (Cys 16.5 nM, Hcy 16.8 nM, GSH 21.3 nM, Cap 26.6 nM). These attributes enabled easy and efficient qualification of Cys on test strips and accurate determination of the effective content of captopril tablets. Notably, PHL-DNP demonstrated low cytotoxicity and precise lysosomal targeting. Through bioimaging, PHL-DNP not only monitored changes in biothiol levels under oxidative stress but also assessed biothiols in complex biological systems such as live HeLa cells, zebrafish, tumor tissue sections, and radish roots. This provides a promising tool for quantitative analysis of biothiols, disease marker detection, and drug testing.

Fine-tuning of the dual-role transcription factor WRKY8 via differential phosphorylation for robust broad-spectrum plant immunity.

Plants utilize plasma membrane-localized pattern recognition receptors (PRRs) to perceive pathogen-associated molecular patterns (PAMPs) to activate broad-spectrum pattern-triggered immunity (PTI). However, the regulatory mechanism ensuring robust broad-spectrum plant immunity remains largely unknown. Here, we reveal the dual roles of the transcription factor WRKY8 in transcriptional regulation of PRR genes: repressing the nlp20/nlp24 receptor gene RLP23 whereas promoting the chitin receptor gene CERK1. Remarkably, SsNLP1 and SsNLP2, two nlp24 type PAMPs in the destructive fungal pathogen Sclerotinia sclerotiorum, activate two calcium-elicited kinases, CPK4 and CPK11 to phosphorylate WRKY8 and consequently release its inhibition on RLP23 expression to accumulate RLP23. Meanwhile, SsNLPs activate a RLCK type kinase, PBL19 to phosphorylate WRKY8 and consequently enhance the accumulation of CERK1. Intriguingly, RLP23 is repressed at late stage by PBL19-mediated phosphorylation of WRKY8, to avoid excessive immunity for normal growth. Our findings unveil a "killing two birds with one stone" strategy employed by plants to elicit robust broad-spectrum immunity, which is based on PAMP-triggered fine-tuning of a dual-role transcription factor to simultaneously amplify two PRRs recognizing PAMPs well conserved in a wide range of pathogens. Moreover, our results reveal a novel plant strategy based on fine-tuning of multiple PRR gene expression to balance the trade-off between growth and immunity.

Identification and Prediction of Differentially Expressed MicroRNAs Associated with Detoxification Pathways in Larvae of Spodoptera frugiperda.

Spodoptera frugiperda poses a severe threat to crops, causing substantial economic losses. The increased use of chemical pesticides has led to resistance in S. frugiperda populations. Micro ribonucleic acids (MicroRNAs or miRNAs) are pivotal in insect growth and development. This study aims to identify miRNAs across different developmental stages of S. frugiperda to explore differential expression and predict target gene functions. High-throughput sequencing of miRNAs was conducted on eggs, 3rd instar larvae, pupae, and adults. Bioinformatics analyses identified differentially expressed miRNAs specifically in larvae, with candidate miRNAs screened to predict target genes, particularly those involved in detoxification pathways. A total of 184 known miRNAs and 209 novel miRNAs were identified across stages. Comparative analysis revealed 54, 15, and 18 miRNAs differentially expressed in larvae, compared to egg, pupa, and adult stages, respectively. Eight miRNAs showed significant differential expression across stages, validated by quantitative reverse transcription PCR (qRT-PCR). Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses predicted target genes' functions, identifying eight differentially expressed miRNAs targeting 10 gene families associated with detoxification metabolism, including P450s, glutathione S-transferase (GSTs), ATP-binding cassette (ABC) transporters, and sodium channels. These findings elucidate the species-specific miRNA profiles and regulatory mechanisms of detoxification-related genes in S. frugiperda larvae, offering insights and strategies for effectively managing this pest.

An improved spectral clustering method for accurate detection of brain resting-state networks.

This paper proposes a data-driven analysis method to accurately partition large-scale resting-state functional brain networks from fMRI data. The method is based on a spectral clustering algorithm and combines eigenvector direction selection with Pearson correlation clustering in the spectral space. The method is an improvement on available spectral clustering methods, capable of robustly identifying active brain networks consistent with those from model-driven methods at different noise levels, even at the noise level of real fMRI data.

Visualizing ClO- fluctuations in drug-induced liver injury and bacterium via a robust ratiometric fluorescent probe.

As a type of reactive oxygen species, hypochlorous acid (ClO-) plays an important role in sterilization, disinfection and protection in organisms. However, excessive production of ClO- is closely related to various diseases. In this work, we have designed a robust ratiometric fluorescent probe, RDB-ClO, using the excited-state intramolecular proton transfer (ESIPT) strategy. RDB-ClO was achieved by modifying 2-(2-(benzo[d]thiazol-2-yl)-6-(diethylamino)-3-oxo-3H-xanthen-9-yl) benzoic acid (RDB-OH) with a 1-naphthoyl chloride group, specifically for the sensitive detection of ClO-. In the presence of ClO-, RDB-ClO demonstrated relatively good performance, showing swift response time (35 s), low detection limit of 5.1 nM and high selectivity towards ClO-. Notably, the convenience and accessibility detection of ClO- has been implemented using test strip and agarose probe. RDB-ClO effectively tracked both endogenous and exogenous ClO- in HeLa cells, HepG2 cells and zebrafish. Additionally, it is successfully applied to detect changes of exogenous ClO- content in E. coli. and acetaminophen (APAP)-induced liver injury in mice. The development of RDB-ClO represents a promising molecular tool for studying the pathogenesis of DILI and biotransformation of ClO- in bacteria.

Radiopaque and Biocompatible PMMA Bone Cement Triggered by Nano Tantalum Carbide and Its Osteogenic Performance.

Poly(methyl methacrylate) (PMMA) bone cements have been widely used in orthopedics; thanks to their excellent mechanical properties, biocompatibility, and chemical stability. Barium sulfate and zirconia are usually added into PMMA bone cement to enhance the X-ray radiopacity, while the mechanical strength, radiopacity, and biocompatibility are not well improved. In this study, an insoluble and corrosion-resistant ceramic, tantalum carbide (TaC), was added into the PMMA bone cement as radiopacifies, significantly improving the mechanical, radiopaque, biocompatibility, and osteogenic performance of bone cement. The TaC-PMMA bone cement with varied TaC contents exhibits compressive strength over 100 MPa, higher than that of the commercial 30% BaSO4-PMMA bone cement. Intriguingly, when the TaC content reaches 20%, the radiopacity is equivalent to the commercial bone cement with 30% of BaSO4 in PMMA. The cytotoxicity and osteogenic performance indicate that the incorporation of TaC not only enhances the osteogenic properties of PMMA but also does not reduce cell viability. This study suggests that TaC could be a superior and multifunctional radio-pacifier for PMMA bone cement, offering a promising avenue for improving patient outcomes in orthopedic applications.

Unlocking a Fast Track for Magnesium Migration in Cu1.8S1-xSex via Anion-Tuning Chemistry.

Rechargeable magnesium batteries (rMBs) are promising candidates for next-generation batteries in which sulfides are widely used as cathode materials. The slow kinetics, low redox reversibility, and poor magnesium storage stability induced by the large Coulombic resistance and ionic polarization of Mg2+ ions have obstructed the development of high-performance rMBs. Herein, a Cu1.8S1-xSex cathode material with a two-dimensional sheet structure has been prepared by an anion-tuning strategy, achieving improved magnesium storage capacity and cycling stability. Element-specific synchrotron radiation analysis is evidence that selenium incorporation has indeed changed the chemical state of Cu species. Density functional theory calculations combined with kinetics analysis reveal that the anionic substitution endows the Cu1.8S1-xSex electrode with favorable charge-transfer kinetics and low ion diffusion barrier. The principal magnesium storage mechanisms and structural evolution process have been revealed in details based on a series of ex situ investigations. Our findings provide an effective heteroatom-tuning tactic of optimizing electrode structure toward advanced energy storage devices.

TACE plus lenvatinib and tislelizumab for intermediate-stage hepatocellular carcinoma beyond up-to-11 criteria: a multicenter cohort study.

Background: Intermediate-stage (BCLC-B) hepatocellular carcinoma (HCC) beyond the up-to-11 criteria represent a significant therapeutic challenge due to high and heterogeneous tumor burden. This study evaluated the effectiveness and safety of transarterial chemoembolization (TACE) in combination with lenvatinib and tislelizumab for these patients. Methods: In this retrospective cohort study, patients with unresectable intermediate-stage HCC beyond the up-to-11 criteria were enrolled and divided into TACE monotherapy (T), TACE combined with lenvatinib (TL), or TACE plus lenvatinib and tislelizumab (TLT) group based on the first-line treatment, respectively. The primary endpoint was overall survival (OS). The secondary outcomes included progression-free survival (PFS), tumor response according to RESIST1.1 and modified RECIST, and adverse events (AEs). Results: There were 38, 45, and 66 patients in the T, TL, and TLT groups, respectively. The TLT group exhibited significantly higher ORR and DCR than the other two groups, as assessed by either mRECIST or RECIST 1.1 (all P<0.05). Median PFS and OS were significantly longer in the TLT group compared with the T group (PFS: 8.5 vs. 4.4 months; OS: 31.5 vs. 18.5 months; all P<0.001) and TL group (PFS: 8.5 vs. 5.5 months; OS: 31.5 vs. 20.5 months; all P<0.05). The incidence of TRAEs was slightly higher in the TLT and TL groups than in the T group, while all the toxicities were tolerable. No treatment-related death occurred in all groups. Conclusions: TACE combined with lenvatinib and tislelizumab significantly improved the survival benefit compared with TACE monotherapy and TACE plus lenvatinib in patients with intermediate-stage HCC beyond the up-to-11 criteria, with an acceptable safety profile.