Host selection by thrips is affected by the floral volatile profile of sunflower.

Thrips, Frankliniella intonsa, is a highly polyphagous pest with a worldwide distribution. F. intonsa-infested sunflower seeds show marked visual damage. The study findings revealed that significantly more F. intonsa infested confection sunflower compared to oilseed sunflower, via olfactometer bioassay studies, we found that compared with the flower and pollen of oilseed sunflowers, those of confection sunflowers attract F. intonsa. Considering this discrepancy in the preference of F. intonsa on oilseed and confection sunflowers, the volatiles of the flower and pollens of two sunflowers were analysed by gas chromatography-mass spectroscopy. The behavioural responses of F. intonsa were assessed for these compounds using Y-tube bioassays. Geranyl bromide, a unique volatile component of oilseed sunflowers, induced an assertive approach-avoidance behaviour in F. intonsa, whereas the unique component ethyl isovalerate in confection sunflowers attracted F. intonsa. F. intonsa adults demonstrated significant attraction to the blends of confection sunflowers. Furthermore, field verification revealed that intercropping confection and oilseed sunflowers could effectively control F. intonsa. The study provided insights into the chemical cues used by F. intonsa in locating hosts. Therefore, oilseed sunflowers can be used as repellent plants to prevent F. intonsa invasion.

Application of the molecular dynamics simulation GROMACS in food science.

Food comprises proteins, lipids, sugars and various other molecules that constitute a multicomponent biological system. It is challenging to investigate microscopic changes in food systems solely by performing conventional experiments. Molecular dynamics (MD) simulation serves as a crucial bridge in addressing this research gap. The Groningen Machine for Chemical Simulations (GROMACS) is an open-source, high-performing molecular dynamics simulation software that plays a significant role in food science research owing to its high flexibility and powerful functionality; it has been used to explore the molecular conformations and the mechanisms of interaction between food molecules at the microcosmic level and to analyze their properties and functions. This review presents the workflow of the GROMACS software and emphasizes the recent developments and achievements in its applications in food science research, thus providing important theoretical guidance and technical support for obtaining an in-depth understanding of the properties and functions of food.

Epstein-Barr virus-positive iris diffuse large B-cell lymphoma detected by metagenomic next-generation sequencing.

PURPOSE: Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) is a relatively rare subtype of DLBCL. Herein, we report a case of a patient with EBV-positive iris DLBCL after undergoing penetrating keratoplasty and discuss its possible pathogenesis. METHODS: A 72-year-old male patient presented to our hospital with progressive blurring of vision in the left eye for the past 4 months. Small white nodular lesions were observed on the iris and retinal surface of the left eye, with a white cloud-like opacity in the vitreous cavity. RESULTS: The patient was eventually diagnosed with EBV-positive iris DLBCL after undergoing pathological and metagenomic tests. After injecting methotrexate in the left vitreous cavity and administering systemic and local antiviral treatments, the ocular lesions disappeared. CONCLUSION: EBV infection, drug immunosuppression, and aging-related immune deterioration may play significant roles in the pathogenesis of EBV-positive iris DLBCL. SYNOPSIS: Epstein-Barr virus (EBV)-positive diffuse large B-cell lymphoma (DLBCL) is a new subtype of DLBCL, which rarely occurs. Herein, we report a case of a patient with EBV-positive iris DLBCL after undergoing penetrating keratoplasty and discuss its possible pathogenesis.

High sensitivity Sagnac interferometric temperature sensor using stress region refractive index modulation.

An all-fiber temperature sensor employing intentional refractive index modulation is experimentally demonstrated. The sensor consists of four sections of polarization maintaining fiber (FPMF) sandwiched between multi-mode fiber (MMF). The stress region of two sections of polarization maintaining fibers (PMFs) is aligned and then anti-clockwise rotates one PMF in 10 deg angles while the other keeps still. Finally, the discharge proceeds. The remaining two PMFs are fusion spliced with the same method. Then the prepared FPMF-MMF structure is connected to the 3 dB coupler to construct a Sagnac loop. The temperature sensitivity reaches 1.49 nm/°C for a temperature range from 16°C to 55°C. The proposed temperature sensor with easy fabrication and good linearity in measuring temperature can be a promising candidate for various applications in environmental monitoring and industrial production.

Adsorption, natural attenuation, and microbial community response of ofloxacin and oxolinic acid in marine sediments.

The pollution of quinolone antibiotics in the marine environment has attracted widespread attention, especially for ofloxacin (OFL) and oxolinic acid (OXO) due to their frequent detection. However, few studies have been conducted to assess the behaviors and microbial community response to these antibiotics in marine sediments, particularly for potential antibiotic-resistant bacteria. In this work, the adsorption characteristics, natural attenuation characteristics, and variation of microbial communities of OFL and OXO in marine sediments were investigated. The adsorption process of antibiotics in sediments occurred on the surface and internal pores of organic matter, where OFL was more likely to be transferred from seawater to sediment compared with OXO. Besides, the adsorption of two antibiotics on sediment surfaces was attributed to physisorption (pore filling, electrostatic interaction) and chemisorption (hydrogen bonding). The natural attenuation of OFL and OXO in marine sediment followed second-order reaction kinetics with half-lives of 6.02 and 26.71 days, respectively, wherein biodegradation contributed the most to attenuation, followed by photolysis. Microbial community structure in marine sediments exposure to antibiotics varied by reducing abundance and diversity of microbial communities, as a whole displaying as an increase in the relative abundance of Firmicutes whereas a decrease of Proteobacteria. In detail, Escherichia-Shigella sp., Blautia sp., Bifidobacterium sp., and Bacillus sp. were those antibiotic-resistant bacteria with potential ability to degrade OFL, while Bacillus sp. may be resistant to OXO. Furthermore, functional predictions indicated that the microbial communities in sediment may resist the stress caused by OFL and OXO through cyano-amino acid metabolism, and ascorbate and aldarate metabolism, respectively. The research is key to understanding fate and bacterial resistance of antibiotics in marine sediments.

Emergency response to ecological protection in maritime phenol spills: Emergency monitor, ecological risk assessment, and reduction.

Recently, hundreds of maritime accidental spills of hazardous chemicals have raised public concerns, especially for phenol due to its potential of spills and highly toxicity. Therefore, for marine ecological protection, this article prepared specific strategies of emergency response to phenol spills. Through the identification for phenol behavior at sea, migration prediction, emergency monitor, as well as their new methods were reviewed. Further, ecological risk assessment and seawater quality criteria were conducted by using a species sensitivity distribution (SSD) approach, wherein, risk quotient (RQ) indicated phenol of simulated marine spills posed a high risk (RQ > 1) in 30 days. The method with eco-friendliness and high-efficiency for phenol reduction was constructed by combination of dredging equipment such as pneumatic dredgers (Airlift) and bioremediation, where marine microorganisms that degraded phenol were summarized, as well as future research needs. This study provided a guidance for emergency response and policy development of phenol spills.

A novel peptide PDHK1-241aa encoded by circPDHK1 promotes ccRCC progression via interacting with PPP1CA to inhibit AKT dephosphorylation and activate the AKT-mTOR signaling pathway.

BACKGROUND: Clear cell renal cell carcinoma (ccRCC) is the most prevalent kidney cancer with high aggressive phenotype and poor prognosis. Accumulating evidence suggests that circRNAs have been identified as pivotal mediators in cancers. However, the role of circRNAs in ccRCC progression remains elusive. METHODS: The differentially expressed circRNAs in 4 paired human ccRCC and adjacent noncancerous tissues ccRCC were screened using circRNA microarrays and the candidate target was selected based on circRNA expression level using weighted gene correlation network analysis (WGCNA) and the gene expression omnibus (GEO) database. CircPDHK1 expression in ccRCC and adjacent noncancerous tissues (n = 148) were evaluated along with clinically relevant information. RT-qPCR, RNase R digestion, and actinomycin D (ActD) stability test were conducted to identify the characteristics of circPDHK1. The subcellular distribution of circPDHK1 was analyzed by subcellular fractionation assay and fluorescence in situ hybridization (FISH). Immunoprecipitation-mass spectrometry (IP-MS) and immunofluorescence (IF) were employed to evaluate the protein-coding ability of circPDHK1. ccRCC cells were transfected with siRNAs, plasmids or lentivirus approach, and cell proliferation, migration and invasion, as well as tumorigenesis and metastasis in nude mice were assessed to clarify the functional roles of circPDHK1 and its encoded peptide PDHK1-241aa. RNA-sequencing, western blot analysis, immunoprecipitation (IP) and chromatin immunoprecipitation (ChIP) assays were further employed to identify the underlying mechanisms regulated by PDHK1-241aa. RESULTS: CircPDHK1 was upregulated in ccRCC tissues and closely related to WHO/ISUP stage, T stage, distant metastasis, VHL mutation and Ki-67 levels. CircPDHK1 had a functional internal ribosome entry site (IRES) and encoded a novel peptide PDHK1-241aa. Functionally, we confirmed that PDHK1-241aa and not the circPDHK1 promoted the proliferation, migration and invasion of ccRCC. Mechanistically, circPDHK1 was activated by HIF-2A at the transcriptional level. PDHK1-241aa was upregulated and interacted with PPP1CA, causing the relocation of PPP1CA to the nucleus. This thereby inhibited AKT dephosphorylation and activated the AKT-mTOR signaling pathway. CONCLUSIONS: Our data indicated that circPDHK1-encoded PDHK1-241aa promotes ccRCC progression by interacting with PPP1CA to inhibit AKT dephosphorylation. This study provides novel insights into the multiplicity of circRNAs and highlights the potential use of circPDHK1 or PDHK1-241aa as a therapeutic target for ccRCC.

RNA-binding protein IGF2BP2 suppresses metastasis of clear cell renal cell carcinoma by enhancing CKB mRNA stability and expression.

Clear cell renal cell carcinoma (ccRCC) is the most prevalent kidney cancer, with a highly aggressive phenotype and poor prognosis. RNA binding proteins (RBPs) play crucial roles in post-transcriptional gene regulation and have been implicated in tumorigenesis. RBPs have the potential to become a new therapeutic target for ccRCC. In this study, we screened and validated that insulin-like growth factor 2 mRNA binding protein 2 (IGF2BP2) as an RBP, was down-regulated in ccRCC tissues and cell lines. Functionally, we verified that IGF2BP2 significantly suppressed the migration and invasion ability of ccRCC in vitro and in vivo. Mechanistically, RIP-seq and actinomycin D experiments results showed that IGF2BP2 enhanced the expression of Creatine Kinase B (CKB) by binding to CKB mRNA and enhancing its mRNA stability. Thus, IGF2BP2 inhibited ccRCC metastasis through enhancing the expression of CKB. Taken together, these finding suggests that IGF2BP2 is a novel metastasis suppressor of ccRCC and may serve as a potential therapeutic target.

RNA Binding Protein PTBP1 Promotes the Metastasis of Gastric Cancer by Stabilizing PGK1 mRNA.

Gastric cancer (GC) is the most common type of malignant tumor within the gastrointestinal tract, and GC metastasis is associated with poor prognosis. Polypyrimidine tract binding protein 1 (PTBP1) is an RNA-binding protein implicated in various types of tumor development and metastasis. However, the role of PTBP1 in GC metastasis remains elusive. In this study, we verified that PTBP1 was upregulated in GC tissues and cell lines, and higher PTBP1 level was associated with poorer prognosis. It was shown that PTBP1 knockdown in vitro inhibited GC cell migration, whereas PTBP1 overexpression promoted the migration of GC cells. In vivo, the knockdown of PTBP1 notably reduced both the size and occurrence of metastatic nodules in a nude mice liver metastasis model. We identified phosphoglycerate kinase 1 (PGK1) as a downstream target of PTBP1 and found that PTBP1 increased the stability of PGK1 by directly binding to its mRNA. Furthermore, the PGK1/SNAIL axis could be required for PTBP1's function in the promotion of GC cell migration. These discoveries suggest that PTBP1 could be a promising therapeutic target for GC.

Effect of ultrahigh pressure processing (UHP) on physicochemical properties, antioxidant activity and anti-inflammatory activity of insoluble dietary fiber from Pholiota nameko.

The aim of this study was to evaluate the effect of ultrahigh pressure processing (UHP) of 200, 300, 400, 500, 600 and 700 MPa for 20, 40 and 30 min on physicochemical and bioactive properties of the insoluble dietary fiber Pholiota nameko (PN-IDF). The results revealed that UHP were capable of decreasing the particle size of PN-IDF and binding phenolic content. Moreover, UHP technique had an improving effect on the bioaccessible phenolic content, the water-holding capacity, the oil-holding capacity and the nitrite ion adsorption capacity. Further, UHP technique presented a promoting effect on the antioxidant activity by scavenging ABTS or DPPH free radicals and increasing reducing power, and the anti-inflammatory activity by inhibiting carrageenan-induced paw edema on PN-IDF. Overall, this study well proved that UHP technology could improve the physicochemical and functional quality of PN-IDF, which could be used as a promising green technique for functional food ingredients processing.

Knowledge mapping of acupoint sensitization and acupoint specificity: a bibliometric analysis.

Objective: The relationship between acupoint sensitization and acupoint specificity is a topic of significant interest in acupuncture research. Numerous clinical studies have demonstrated that needling pain sensitive acupoints yields superior therapeutic outcomes compared to traditional acupoints, particularly in the context of pain disorders. However, there is a lack of bibliometric analysis in acupuncture area. Therefore, the objective of this study is to offer a comprehensive overview of the knowledge structure and research hotspots pertaining to acupoint sensitization and acupoint specificity. Methods: The search for publications pertaining to acupoint sensitization and acupoint specificity was conducted in the Web of Science Core Collection (WoSCC) database from its inception until August 11, 2023. Subsequently, bibliometric analyses were carried out using VOSviewer, CiteSpace, R software (Bibliometrix package), and GraphPad Prism software. Results: This study includes 4,940 articles from 72 countries, with China and the United States being the leading countries. The number of publications related to acupoint sensitization and specificity has been increasing annually. Major research institutions involved in this field include the Shanghai University of Traditional Chinese Medicine, Kyung Hee University, Beijing University of Chinese Medicine, Chinese Academy of Chinese Medical Sciences, and China Medical University, among others. "Evidence-based Complementary and Alternative Medicine" is the most popular journal in acupuncture field, and "PAIN" is the most co-cited journal. Publications are contributed by 20,325 authors from all over the world, with Wu Huangan, Fang Jianqiao, Lin Yi-Wen, Liu Huirong, and Chen Rixin having published the most articles. Han Ji-Sheng is the most cited author in this research area. The main directions include the study of temperature specificity of acupoints, the diagnosis of acupoint sensitization diseases, and the study of the mechanism of acupoint sensitization. The most listed keywords in recent years are "TRPV1," "signaling pathway," and "diagnosis." Conclusion: This is the first bibliometric study to comprehensively summarize research trends and advances in acupoint sensitization and acupoint specificity, and the information highlights recent research preliminary and main directions that serve as a reference for acupoint sensitization and acupoint specificity research.

Evaluation of the efficacy and safety of CalliSpheres® microsphere-transarterial chemoembolization in large hepatocellular carcinoma.

OBJECTIVE: The prognosis of large hepatocellular carcinoma (HCC) is still unfavorable due to limited and challenging treatment. CalliSpheres® microsphere-transarterial chemoembolization (CSM-TACE) is an effective therapy for general HCC but not frequently applied for large HCC. Hence, this study aimed to investigate the efficacy and safety of CSM-TACE in large HCC patients. MATERIALS AND METHODS: This prospective study analyzed 100 large HCC (tumor size >5 cm) patients receiving CSM-TACE. Treatment response, survival, change in liver function indexes, and adverse events were recorded. RESULT: The best complete response, partial response, stable disease, and progressive disease rates were 2.0%, 31.3%, 65.7%, and 1.0%, respectively, leading to the best objective response rate (ORR) of 33.3% and disease control rate of 99.9%. Multivariate analysis showed that intrahepatic metastasis was independently related to poor ORR (odd ratio = 0.366, P = 0.023). The 1- and 2-year progression-free survival (PFS) rates were 88.9% and 80.6%, with a mean [95% confidence interval (CI)] PFS of 21.6 (20.4-22.9) months. The 1- and 2-year overall survival (OS) rates were 99.0% and 99.0%, with a mean (95% CI) OS of 23.8 (23.3-24.2) months. Total bilirubin (P < 0.001), alanine transaminase (P < 0.001), aspartate transaminase (P < 0.001), and α-fetoprotein (P = 0.045) were abnormal in a short-term period then stably recovered from 1 month ± 15 days after drug-eluting bead-TACE to 24 months ± 15 days. During hospitalization and postdischarge, tolerable abdominal pain and decreased appetite were common adverse events. CONCLUSIONS: CSM-TACE shows favorable treatment response and survival with acceptable tolerance among large HCC patients, indicating that it may promote the management of these patients.

Stability and Crystallinity of Sodium Poly(Heptazine Imide) in Photocatalysis.

Poly(heptazine imide) (PHI) salts, as crystalline carbon nitrides, exhibit high photocatalytic activity and are being extensively researched, but its photochemical instability has not drawn researchers' attention yet. Herein, sodium PHI (PHI-Na) ultrathin nanosheets with increased crystallinity, synthesized by enhancing contact of melamine with NaCl functioning as a structure-induction agent and hard template, exhibits improved photocatalytic hydrogen evolution activity, but low photochemical stability, owing to Na+ loss in the photocatalytic process, which, interestingly, can be enhanced by the common ion effect, e.g., addition of NaCl that is also able to remarkably increase the photoactivity with the apparent quantum yield at 420 nm reaching 41.5 %. This work aims at attracting research peers' attention to photochemical instability of PHI salts and provides a way to enhance their crystallinity.

Efficiency, mechanism, influencing factors, and integrated technology of biodegradation for aromatic compounds by microalgae: A review.

Aromatic compounds have received widespread attention because of their threat to ecosystem and human health. However, traditional physical and chemical methods are criticized due to secondary pollution and high cost. As a result of ecological security and the ability of carbon sequestration, biodegradation approach based on microalgae has emerged as a promising alternative treatment for aromatic pollutants. In light of the current researches, the degradation efficiency of BTEX (benzene, toluene, ethylbenzene, and xylene), polycyclic aromatic hydrocarbons (PAHs), and phenolic compounds by microalgae was reviewed in this study. We summarized the degradation pathways and metabolites of p-xylene, benzo [a]pyrene, fluorene, phenol, bisphenol A, and nonylphenol by microalgae. The influence factors on the degradation of aromatic compounds by microalgae were also discussed. The integrated technologies based on microalgae for degradation of aromatic compounds were reviewed. Finally, this study discussed the limitations and future research needs of the degradation of these compounds by microalgae.

Interfacial Affinity Determined Photocatalytic Activity: A Comparison between Defective and Bulk Polymeric Carbon Nitride.

The photoexcited charge separation efficiency of photocatalysts is generally considered as the key factor for enhancement of their photocatalytic activity, and sometimes, their photoabsorption capability and interfacial reaction kinetics play a key part, but the role of interfacial affinity of photocatalysts with substrates was rarely researched systematically. Herein, nitrogen vacancy-modified polymeric carbon nitride porous nanotubes (PCNpts) were simply synthesized, using tartaric acid as a crosslinking and corrosion agent, and exhibit a remarkable increment in surface area, wettability, photoabsorption and charge separation capability, and photocatalytic activity in water splitting to produce H2, but, interestingly, exhibit substrate-dependent variation of photoactivity in contaminant degradation, compared with bulk PCN. More interestingly, the interfacial affinity of PCNpts and PCN with contaminants and H2O, rather than photoabsorption and charge separation capability, is confirmed to dominate their photoactivity.

CircNFATC3 promotes the proliferation of gastric cancer through binding to IGF2BP3 and restricting its ubiquitination to enhance CCND1 mRNA stability.

BACKGROUND: Insulin like growth factor II mRNA binding protein 3 (IGF2BP3) is an RNA binding protein with multiple roles in regulation of gene expression at the post-transcriptional level and is implicated in tumorigenesis and progression of numerous cancers including gastric cancer (GC). Circular RNAs (circRNAs) are a diverse endogenous noncoding RNA population that have important regulatory roles in cancer. However, circRNAs that regulate the expression of IGF2BP3 in GC is largely unknown. METHODS: CircRNAs that bound to IGF2BP3 were screened in GC cells using RNA immunoprecipitation and sequencing (RIP-seq). The identification and localization of circular nuclear factor of activated T cells 3 (circNFATC3) were identified using Sanger sequencing, RNase R assays, qRT-PCR, nuclear-cytoplasmic fractionation and RNA-FISH assays. CircNFATC3 expression in human GC tissues and adjacent normal tissues were measured by qRT-PCR and ISH. The biological role of circNFATC3 in GC was confirmed by in vivo and in vitro experiments. Furthermore, RIP, RNA-FISH/IF, IP and rescue experiments were performed to uncover interactions between circNFATC3, IGF2BP3 and cyclin D1 (CCND1). RESULTS: We identified a GC-associated circRNA, circNFATC3, that interacted with IGF2BP3. CircNFATC3 was significantly overexpressed in GC tissues and was positively associated with tumor volume. Functionally, the proliferation of GC cells decreased significantly after circNFATC3 knockdown in vivo and in vitro. Mechanistically, circNFATC3 bound to IGF2BP3 in the cytoplasm, which enhanced the stability of IGF2BP3 by preventing ubiquitin E3 ligase TRIM25-mediated ubiquitination, thereby enhancing the regulatory axis of IGF2BP3-CCND1 and promoting CCND1 mRNA stability. CONCLUSIONS: Our findings demonstrate that circNFATC3 promotes GC proliferation by stabilizing IGF2BP3 protein to enhance CCND1 mRNA stability. Therefore, circNFATC3 is a potential novel target for the treatment of GC.

Ecological risk assessment for xylenes and propylbenzenes in aquatic environment using a species sensitivity distribution approach.

Xylenes and propylbenzenes (PBZs) are volatile aromatic hydrocarbons with high aquatic toxicity. Xylenes can be present in three isomers: o-xylene (OX), m-xylene (MX), and p-xylene (PX), while PBZs include two isomers: n-propylbenzene (n-PBZ) and isopropylbenzene (i-PBZ). Their accidental spills and improper discharges from petrochemical industries can cause severe contamination in water bodies posing potential ecological risks. In this study, the published acute toxicity data of these chemicals for aquatic species were collected to calculate hazardous concentrations protecting 95% species (HC5) using a species sensitivity distribution (SSD) approach. The acute HC5 values for OX, MX, PX, n-PBZ, and i-PBZ were estimated to be 1.73, 3.05, 1.23, 1.22, and 1.46 mg/L, respectively. The risk quotient (RQ) values calculated based on HC5 indicated their high risk (RQ: 1.23 ∼ 21.89) in groundwater, but low risk (RQ < 0.1) in natural seawater, river water, and lake water. When xylenes or PBZs leaked into the sea, they were expected to pose a high risk (RQ > 1) at the start and then a low risk (RQ < 0.1) after 10 days due to natural attenuation. These results may help to derive more reliable protection thresholds for xylenes and PBZs in aquatic environment and provide a basis for evaluating their ecological risks.

Detection of methyltransferase activity and inhibitor screening based on rGO-mediated silver enhancement signal amplification strategy.

DNA methylation refers to the chemical modification process of obtaining a methyl group by the covalent bonding of a specific base in DNA sequence with S-adenosyl methionine (SAM) as a methyl donor under the catalysis of methyltransferase (MTase), which is related to the occurrence of multiple diseases. Therefore, the detection of MTase activity is of great significance for disease diagnosis and drug screening. Because reduced graphene oxide (rGO) has a unique planar structure and remarkable catalytic performance, it is not clear whether rGO can rapidly catalyze silver deposition as an effective way of signal amplification. However, in this study, we were pleasantly surprised to find that using H2O2 as a reducing agent, rGO can rapidly catalyze silver deposition, and its catalytic efficiency of silver deposition is significantly better than that of GO. Therefore, based on further verifying the mechanism of catalytic properties of rGO, we constructed a novel electrochemical biosensor (rGO/silver biosensor) for the detection of dam MTase activity, which has high selectivity and sensitivity to MTase in the range of 0.1 U/mL to 10.0 U/mL, and the detection limit is as low as 0.07 U/mL. Besides, this study also used Gentamicin and 5-Fluorouracil as inhibitor models, confirming that the biosensor has a good application prospect in the high-throughput screening of dam MTase inhibitors.

A Phase 1/2 Multicenter Randomized Trial of Local Ablation plus Toripalimab versus Toripalimab Alone for Previously Treated Unresectable Hepatocellular Carcinoma.

PURPOSE: To assess the safety and efficacy of local ablation plus PD-1 inhibitor toripalimab in previously treated unresectable hepatocellular carcinoma (HCC). PATIENTS AND METHODS: In the multicenter, two-stage, and randomized phase 1/2 trial, patients were randomly assigned to receive toripalimab alone (240 mg, every 3 weeks), subtotal local ablation followed by toripalimab starting on post-ablation day 3 (Schedule D3), or on post-ablation day 14 (Schedule D14). The first endpoint of stage 1 was to determine which combination schedule could continue and progression-free survival (PFS) as the primary endpoint for stage 1/2. RESULTS: A total of 146 patients were recruited. During stage 1, Schedule D3 achieved numerically higher objective response rate (ORR) than Schedule D14 for non-ablation lesions (37.5% vs. 31.3%), and was chosen for stage 2 evaluation. For the entire cohort of both stages, patients with Schedule D3 had a significantly higher ORR than with toripalimab alone (33.8% vs. 16.9%; P = 0.027). Moreover, patients with Schedule D3 had improved median PFS (7.1 vs. 3.8 months; P < 0.001) and median overall survival (18.4 vs. 13.2 months; P = 0.005), as compared with toripalimab alone. In addition, six (9%) patients with toripalimab, eight (12%) with Schedule D3, and 4 (25%) with Schedule D14 developed grade 3 or 4 adverse events, and one patient (2%) with Schedule D3 manifested grade 5 treatment-related pneumonitis. CONCLUSIONS: In patients with previously treated unresectable HCC, subtotal ablation plus toripalimab improved the clinical efficacy as compared with toripalimab alone, with an acceptable safety profile.