Low-intensity transcranial focused ultrasound suppresses pain by modulating pain-processing brain circuits.

ABSTRACT: There is an urgent and unmet clinical need to develop nonpharmacological interventions for chronic pain management because of the critical side effects of opioids. Low-intensity transcranial focused ultrasound (tFUS) is an emerging noninvasive neuromodulation technology with high spatial specificity and deep brain penetration. Here, we developed a tightly focused 128-element ultrasound transducer to specifically target small mouse brains using dynamic focus steering. We demonstrate that tFUS stimulation at pain-processing brain circuits can significantly alter pain-associated behaviors in mouse models in vivo. Our findings indicate that a single-session focused ultrasound stimulation to the primary somatosensory cortex (S1) significantly attenuates heat pain sensitivity in wild-type mice and modulates heat and mechanical hyperalgesia in a humanized mouse model of chronic pain in sickle cell disease. Results further revealed a sustained behavioral change associated with heat hypersensitivity by targeting deeper cortical structures (eg, insula) and multisession focused ultrasound stimulation to S1 and insula. Analyses of brain electrical rhythms through electroencephalography demonstrated a significant change in noxious heat hypersensitivity-related and chronic hyperalgesia-associated neural signals after focused ultrasound treatment. Validation of efficacy was carried out through control experiments, tuning ultrasound parameters, adjusting interexperiment intervals, and investigating effects on age, sex, and genotype in a head-fixed awake model. Importantly, tFUS was found to be safe, causing no adverse effects on motor function or the brain's neuropathology. In conclusion, the validated proof-of-principle experimental evidence demonstrates the translational potential of novel focused ultrasound neuromodulation for next-generation pain treatment without adverse effects.

Heme-induced loss of renovascular endothelial protein C receptor promotes chronic kidney disease in sickle mice.

ABSTRACT: Chronic kidney disease (CKD) is a major contributor to morbidity and mortality in sickle cell disease (SCD). Anemia, induced by chronic persistent hemolysis, is associated with the progressive deterioration of renal health, resulting in CKD. Moreover, patients with SCD experience acute kidney injury (AKI), a risk factor for CKD, often during vaso-occlusive crisis associated with acute intravascular hemolysis. However, the mechanisms of hemolysis-driven pathogenesis of the AKI-to-CKD transition in SCD remain elusive. Here, we investigated the role of increased renovascular rarefaction and the resulting substantial loss of the vascular endothelial protein C receptor (EPCR) in the progressive deterioration of renal function in transgenic SCD mice. Multiple hemolytic events raised circulating levels of soluble EPCR (sEPCR), indicating loss of EPCR from the cell surface. Using bone marrow transplantation and super-resolution ultrasound imaging, we demonstrated that SCD mice overexpressing EPCR were protective against heme-induced CKD development. In a cohort of patients with SCD, plasma sEPCR was significantly higher in individuals with CKD than in those without CKD. This study concludes that multiple hemolytic events may trigger CKD in SCD through the gradual loss of renovascular EPCR. Thus, the restoration of EPCR may be a therapeutic target, and plasma sEPCR can be developed as a prognostic marker for sickle CKD.

Reducing crosstalk of a multi-plane holographic display by the time-multiplexing stochastic gradient descent.

Multi-plane reconstruction is essential for realizing a holographic three-dimensional (3D) display. One fundamental issue in conventional multi-plane Gerchberg-Saxton (GS) algorithm is the inter-plane crosstalk, mainly caused by the neglect of other planes' interference in the process of amplitude replacement at each object plane. In this paper, we proposed the time-multiplexing stochastic gradient descent (TM-SGD) optimization algorithm to reduce the multi-plane reconstruction crosstalk. First, the global optimization feature of stochastic gradient descent (SGD) was utilized to reduce the inter-plane crosstalk. However, the crosstalk optimization effect would degrade as the number of object planes increases, due to the imbalance between input and output information. Thus, we further introduced the time-multiplexing strategy into both the iteration and reconstruction process of multi-plane SGD to increase input information. In TM-SGD, multiple sub-holograms are obtained through multi-loop iteration and then sequentially refreshed on spatial light modulator (SLM). The optimization condition between the holograms and the object planes converts from one-to-many to many-to-many, improving the optimization of inter-plane crosstalk. During the persistence of vision, multiple sub-hologram jointly reconstruct the crosstalk-free multi-plane images. Through simulation and experiment, we confirmed that TM-SGD could effectively reduce the inter-plane crosstalk and improve image quality.The proposed TM-SGD-based holographic display has wide applications in tomographic 3D visualization for biology, medical science, and engineering design, which need to reconstruct multiple independent tomographic images without inter-plane crosstalk.

High-brightness hybrid compressive light field display with improved image quality.

Previous LCD-based multiplicative compressive light field (CLF) display has the trade-off between the brightness and the depth of field (DOF). In this paper, we propose a hybrid CLF display using a reflective polarizer and RGB mini-LED panel. By the polarization-multiplexing and the reflector dam (RD) designed on the mini-LED panel, the proposed system can preserve high brightness while enhancing the DOF. Then, a decomposition algorithm is proposed to improve the image quality by depth segmentation and limiting the motion parallax. Compared to the conventional hybrid CLF display, the brightness of the proposed system reaches 348 nits and the reconstruction quality achieves structural similarity index measure (SSIM) improvement by 0.12. The experiments also demonstrate that the proposed method could achieve a higher brightness, larger depth of field, and higher image quality.

Transcriptomic Analysis and the Effect of Maturity Stage on Fruit Quality Reveal the Importance of the L-Galactose Pathway in the Ascorbate Biosynthesis of Hardy Kiwifruit (Actinidia arguta).

Hardy kiwifruit (Actinidia arguta) has recently become popular in fresh markets due to its edible skin and rich nutritional value. In the present study, different harvest stages of two A. arguta cultivars, 'Issai' and 'Ananasnaya' ("Ana"), were chosen for investigating the effects of maturity on the quality of the fruit. Interestingly, Issai contained 3.34 folds higher ascorbic acid (AsA) content than Ana. The HPLC method was used to determine the AsA content of the two varieties and revealed that Issai had the higher content of AsA and DHA. Moreover, RNA sequencing (RNAseq) of the transcriptome-based expression analysis showed that 30 differential genes for ascorbate metabolic pathways were screened in Issai compared to Ana, which had 16 genes down-regulated and 14 genes up-regulated, while compared to the up-regulation of 8 transcripts encoding the key enzymes involved in the L-galactose biosynthesis pathway. Our results suggested that AsA was synthesized mainly through the L-galactose pathway in hardy kiwifruit.

Current Development and Applications of Super-Resolution Ultrasound Imaging.

Abnormal changes of the microvasculature are reported to be key evidence of the development of several critical diseases, including cancer, progressive kidney disease, and atherosclerotic plaque. Super-resolution ultrasound imaging is an emerging technology that can identify the microvasculature noninvasively, with unprecedented spatial resolution beyond the acoustic diffraction limit. Therefore, it is a promising approach for diagnosing and monitoring the development of diseases. In this review, we introduce current super-resolution ultrasound imaging approaches and their preclinical applications on different animals and disease models. Future directions and challenges to overcome for clinical translations are also discussed.

A WRKY Transcription Factor, EjWRKY17, from Eriobotrya japonica Enhances Drought Tolerance in Transgenic Arabidopsis.

The WRKY gene family, which is one of the largest transcription factor (TF) families, plays an important role in numerous aspects of plant growth and development, especially in various stress responses. However, the functional roles of the WRKY gene family in loquat are relatively unknown. In this study, a novel WRKY gene, EjWRKY17, was characterized from Eriobotrya japonica, which was significantly upregulated in leaves by melatonin treatment during drought stress. The EjWRKY17 protein, belonging to group II of the WRKY family, was localized in the nucleus. The results indicated that overexpression of EjWRKY17 increased cotyledon greening and root elongation in transgenic Arabidopsis lines under abscisic acid (ABA) treatment. Meanwhile, overexpression of EjWRKY17 led to enhanced drought tolerance in transgenic lines, which was supported by the lower water loss, limited electrolyte leakage, and lower levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Further investigations showed that overexpression of EjWRKY17 promoted ABA-mediated stomatal closure and remarkably up-regulated ABA biosynthesis and stress-related gene expression in transgenic lines under drought stress. Overall, our findings reveal that EjWRKY17 possibly acts as a positive regulator in ABA-regulated drought tolerance.

Ultrasound super-resolution imaging provides a noninvasive assessment of renal microvasculature changes during mouse acute kidney injury.

Acute kidney injury (AKI) is a risk factor for the development of chronic kidney disease (CKD). One mechanism for this phenomenon is renal microvascular rarefaction and subsequent chronic impairment in perfusion. However, diagnostic tools to monitor the renal microvasculature in a noninvasive and quantitative manner are still lacking. Ultrasound super-resolution imaging is an emerging technology that can identify microvessels with unprecedented resolution. Here, we applied this imaging technique to identify microvessels in the unilateral ischemia-reperfusion injury mouse model of AKI-to-CKD progression in vivo. Kidneys from 21 and 42 day post- ischemia-reperfusion injury, the contralateral uninjured kidneys, and kidneys from sham-operated mice were examined by ultrasound super-resolution and histology. Renal microvessels were successfully identified by this imaging modality with a resolution down to 32 µm. Renal fibrosis was observed in all kidneys with ischemia-reperfusion injury and was associated with a significant reduction in kidney size, cortical thickness, relative blood volume, and microvascular density as assessed by this imaging. Tortuosity of the cortical microvasculature was also significantly increased at 42 days compared to sham. These vessel density measurements correlated significantly with CD31 immunohistochemistry (R2=0.77). Thus, ultrasound super-resolution imaging provides unprecedented resolution and is capable of noninvasive quantification of renal vasculature changes associated with AKI-to-CKD progression in mice. Hence, this technique could be a promising diagnostic tool for monitoring progressive kidney disease.

Combing superb microvascular imaging with shear wave elastography for risk stratification of Thyroid Imaging Reporting and Data System (TI-RADS) 4 thyroid nodules.

Background: It is difficult to accurately assess the risk of Thyroid Imaging Reporting and Data System (TI-RADS) 4 thyroid nodules due to the overlap of benign and malignant conventional ultrasound (US) features of nodules. To reduce unnecessary needle biopsies and assist clinical decision-making, this study established a dynamic nomogram incorporating superb microvascular imaging (SMI) and shear wave elastography (SWE) for the risk evaluation of TI-RADS 4 thyroid nodules. Methods: A total of 248 patients who underwent US, SMI, and SWE with cytological or histopathological results were included in this retrospective study, and were randomly divided into training (174 patients) and verification (74 patients) cohorts. The clinical characteristics and US, SMI, and SWE features of patients were analyzed in the training cohort. The least absolute shrinkage and selection operator (LASSO) regression and multivariate logistic regression were used to screen parameters and construct dynamic nomogram. The receiver operating characteristic (ROC) curves, calibration curve, and decision curve were used to evaluate the performance of the nomogram. Results: A dynamic nomogram was constructed based on age [odds ratio (OR) =0.954; P=0.005] , shape (OR =0.345; P=0.041), SMI (OR =9.511; P<0.001), and SWE (OR =3.670; P=0.001). The nomogram showed excellent discrimination both in the training [area under the curve (AUC): 0.848; 95% confidence interval (CI): 0.784-0.911] and validation (AUC: 0.862; 95% CI: 0.780-0.944) cohorts, and better than US, SMI, and SWE alone in all cohorts (P<0.05). The Nomo-score of each patient was calculated and the cut-off value was 0.607 which can be used to distinguish high-risk and low-risk patients. Conclusions: The SMI and SWE show added predictive value on risk stratification in patients with TI-RADS 4 thyroid nodules and a dynamic nomogram was constructed to screen high-risk individuals and assist the clinical decision-making.

Polymethoxylated flavone variations and in vitro biological activities of locally cultivated Citrus varieties in China.

Citrus peels are rich in polymethoxylated flavones (PMFs), which have beneficial health and pharmacological properties. In this study, the profiles, variations, and biological activities of PMFs in the peel extracts of 27 Citrus varieties (eight species) native to China were investigated. UPLC-QTOF-MS/MS analysis revealed that mandarin accumulated more diversity and higher detectable PMF contents. Wangcangzhoupigan (ZPG) possessed the highest antioxidant capacity. Gailiangcheng (GLC) and Bingtangcheng (BTC), sweet oranges showed excellent inhibitory effects against pancreatic lipase and α-glucosidase, respectively. Most citrus extracts effectively inhibited the production of ROS and pro-inflammatory cytokines, while increasing the accumulation of anti-inflammatory cytokines. In addition, Limeng (LM), Cupig-oushigan (GSG), and Yanxiwanlu (YXWL) showed anti-proliferative effects against DU145 and PC3 cancer cells. This study provides a comprehensive PMF profile and biological activities of various citrus species and will benefit future functional citrus breeding practices aimed at designing plants rich in total or specific PMFs for health benefits.

3-D real-time ultrasound tracking of acoustically actuated swimming microdrone.

Maneuverable microswimmers/microdrones that navigate in hard-to-reach spaces inside human bodies hold a great potential for various biomedical applications. Acoustically actuated microswimmers have already demonstrated feasibility. However, for eventual translation of this technology, a robust 3-D tracking strategy for the microswimmer is particularly required. This paper presents our lab-designed 3-D ultrasound tracking system for real-time tracking of an acoustically actuated 3-D swimming microdrone. The ultrasound tracking system utilizing two ultrasound probes, a step motor and a host controller, was built to track the 3-D arbitrary motion of the microdrone in real-time. The performance of tracking was evaluated in the benchtop experiments by comparing the reconstructed trajectories with synchronized camera recordings. The ultrasound tracking system showed high reliability, with an average error of less than 0.3 mm across six different trials when compared to camera tracking. The results demonstrated the capability of our lab-designed 3-D ultrasound tracking system in accurately tracking the undetermined motion of the acoustic actuated 3-D swimming microdrone in real-time. The developed tracking system holds promise as a potential approach for biomedical applications and could pave the way for future clinical translation of the microswimmer technology.

Correlation between computed tomography-based body composition parameters and hepatic venous pressure gradient in patients with cirrhosis: a systematic review and meta-analysis.

PURPOSE: Computed tomography (CT)-based body composition parameters and the hepatic venous pressure gradient (HVPG) are key characteristics in patients with liver cirrhosis. The present study aims to explore the correlation between CT-based body composition parameters and HVPG, as well as the difference in HVPG between patients with and patients without sarcopenia. METHODS: A literature search for studies reporting the correlation between HVPG and CT-based body composition parameters published in English up to August 2023 in four databases, Embase, MEDLINE (via PubMed), Web of Science, and Cochrane Library, was conducted. The correlation coefficient between HVPG and CT-based body composition parameters was the primary outcome, and the difference in the HVPG value between the sarcopenia and non-sarcopenia groups was the secondary outcome. A meta-analysis was conducted using a random-effects models. The methodologic quality was assessed using the Quality Assessment of Diagnostic Studies instrument. RESULTS: A total of 652 articles were identified, of which nine studies (n = 1,569) met the eligibility criteria. Among them, seven studies reported the primary outcome via the muscle index, five via the skeletal muscle index (SMI), two via the psoas-muscle-related index (PRI), and three via two adipose tissue indexes. A total of five studies reported the secondary outcome: four via SMI and one via PRI. No evidence of a significant correlation was determined between the various body composition parameters and the HVPG value, either in the muscle index or the adipose tissue index. Higher HVPG values were observed in patients with sarcopenia than in patients without sarcopenia [pooled standardized mean difference (SMD): 0.628 (-0.350, 1.606), P < 0.001; I2 = 92.8%; P < 0.001] when an Asian sarcopenia definition was adopted. In contrast, when a Western cut-off value was applied, the HVPG value was higher in patients without sarcopenia than in patients with sarcopenia [pooled SMD: -0.201 (-0.366, -0.037), P = 0.016; I2 = 0.00%; P = 0.785]. CONCLUSION: No sufficient evidence regarding a correlation between the CT-based body composition and HVPG value was discovered. The difference in the HVPG value between the sarcopenia and non-sarcopenia groups was likely dependent on the sarcopenic cut-off value.

Dual-crosslinking gelatin-hyaluronic acid methacrylate based biomimetic PDAC desmoplastic niche enhances tumor-associated macrophages recruitment and M2-like polarization.

The tumor microenvironment (TME) of pancreatic ductal adenocarcinoma (PDAC) is characterized by deposition of desmoplastic matrix (including collagen and hyaluronic acid). And the interactions between tumor-associated macrophages (TAMs) and tumor cells play a crucial role in progression of PDAC. Hence, the appropriate model of tumor cell-macrophage interaction within the unique PDAC TME is of significantly important. To this end, a 3D tumor niche based on dual-crosslinking gelatin methacrylate and hyaluronic acid methacrylate hydrogels was constructed to simulate the desmoplastic tumor matrix with matching compressive modulus and composition. The bionic 3D tumor niche creates an immunosuppressive microenvironment characterized by the downregulation of M1 markers and upregulation of M2 markers in TAMs. Mechanistically, RNA-seq analysis revealed that the PI3K-AKT signaling pathway might modulate the phenotypic balance and recruitment of macrophages through regulating SELE and VCAM-1. Furthermore, GO and GSEA revealed the biological process of leukocyte migration and the activation of cytokine-associated signaling were involved. Finally, the 3D tumor-macrophage niches with three different ratios were fabricated which displayed increased M2-like polarization and stemness. The utilization of the 3D tumor niche has the potential to provide a more accurate investigation of the interplay between PDAC tumor cells and macrophages within an in vivo setting.

Endothelial Cells Promote Pseudo-islet Function Through BTC-EGFR-JAK/STAT Signaling Pathways.

Interactions between cells are of fundamental importance in affecting cell function. In vivo, endothelial cells and islet cells are close to each other, which makes endothelial cells essential for islet cell development and maintenance of islet cell function. We used endothelial cells to construct 3D pseudo-islets, which demonstrated better glucose regulation and greater insulin secretion compared to conventional pseudo-islets in both in vivo and in vitro trials. However, the underlying mechanism of how endothelial cells promote beta cell function localized within islets is still unknown. We performed transcriptomic sequencing, differential gene analysis, and enrichment analysis on two types of pseudo-islets to show that endothelial cells can promote the function of internal beta cells in pseudo-islets through the BTC-EGFR-JAK/STAT signaling pathway. Min6 cells secreted additional BTC after co-culture of endothelial cells with MIN6 cells outside the body. After BTC knockout in vitro, we found that beta cells functioned differently: insulin secretion levels decreased significantly, while the expression of key proteins in the EGFR-mediated JAK/STAT signaling pathway simultaneously decreased, further confirming our results. Through our experiments, we elucidate the molecular mechanisms by which endothelial cells maintain islet function in vitro, which provides a theoretical basis for the construction of pseudo-islets and islet cell transplants for the treatment of diabetes mellitus.

GMMID: genetically modified mice information database.

Genetically engineered mouse models (GEMMs) are vital for elucidating gene function and disease mechanisms. An overwhelming number of GEMM lines have been generated, but endeavors to collect and organize the information of these GEMMs are seriously lagging behind. Only a few databases are developed for the information of current GEMMs, and these databases lack biological descriptions of allele compositions, which poses a challenge for nonexperts in mouse genetics to interpret the genetic information of these mice. Moreover, these databases usually do not provide information on human diseases related to the GEMM, which hinders the dissemination of the insights the GEMM provides as a human disease model. To address these issues, we developed an algorithm to annotate all the allele compositions that have been reported with Python programming and have developed the genetically modified mice information database (GMMID; http://www.gmmid.cn), a user-friendly database that integrates information on GEMMs and related diseases from various databases, including National Center for Biotechnology Information, Mouse Genome Informatics, Online Mendelian Inheritance in Man, International Mouse Phenotyping Consortium, and Jax lab. GMMID provides comprehensive genetic information on >70 055 alleles, 65 520 allele compositions, and ∼4000 diseases, along with biologically meaningful descriptions of alleles and allele combinations. Furthermore, it provides spatiotemporal visualization of anatomical tissues mentioned in these descriptions, shown alongside the allele compositions. Compared to existing mouse databases, GMMID considers the needs of researchers across different disciplines and presents obscure genetic information in an intuitive and easy-to-understand format. It facilitates users in obtaining complete genetic information more efficiently, making it an essential resource for cross-disciplinary researchers. Database URL: http://www.gmmid.cn.

Dynamic Changes in Flavor and Microbiota in Traditionally Fermented Bamboo Shoots (Chimonobambusa szechuanensis (Rendle) Keng f.).

Dissecting flavor formation and microbial succession during traditional fermentation help to promote standardized and large-scale production in the sour shoot industry. The principal objective of the present research is to elucidate the interplay between the physicochemical attributes, flavor, and microbial compositions of sour bamboo shoots in the process of fermentation. The findings obtained from the principal component analysis (PCA) indicated notable fluctuations in both the physicochemical parameters and flavor components throughout the 28 day fermentation process. At least 13 volatile compounds (OAV > 1) have been detected as characteristic aroma compounds in sour bamboo shoots. Among these, 2,4-dimethyl Benzaldehyde exhibits the highest OAV (129.73~668.84) and is likely the primary contributor to the sour odor of the bamboo shoots. The analysis of the microbial community in sour bamboo shoots revealed that the most abundant phyla were Firmicutes and Proteobacteria, while the most prevalent genera were Enterococcus, Lactococcus, and Serratia. The results of the correlation analysis revealed that Firmicutes exhibited a positive correlation with various chemical compounds, including 3,6-nonylidene-1-ol, 2,4-dimethyl benzaldehyde, silanediol, dimethyl-, nonanal, and 2,2,4-trimethyl-1,3-pentylenediol diisobutyrate. Similarly, Lactococcus was found to be positively correlated with several chemical compounds, such as dimethyl-silanediol, 1-heptanol, 3,6-nonylidene-1-ol, nonanal, 2,2,4-trimethyl-1,3-pentanediol diisobutyrate, dibutyl phthalate, and TA. This study provides a theoretical basis for the standardization of traditional natural fermented sour bamboo production technology, which will help to further improve the flavor and quality of sour bamboo.

Depth-Enhanced Holographic Super Multi-View Maxwellian Display Based on Variable Filter Aperture.

The super multi-view (SMV) near-eye display (NED) effectively provides depth cues for three-dimensional (3D) displays by projecting multiple viewpoint images or parallax images onto the retina simultaneously. Previous SMV NED suffers from a limited depth of field (DOF) due to the fixed image plane. Aperture filtering is widely used to enhance the DOF; however, an invariably sized aperture may have opposite effects on objects with different reconstruction depths. In this paper, a holographic SMV display based on the variable filter aperture is proposed to enhance the DOF. In parallax image acquisition, multiple groups of parallax images, each group recording a part of the 3D scene on a fixed depth range, are captured first. In the hologram calculation, each group of wavefronts at the image recording plane (IRP) is calculated by multiplying the parallax images with the corresponding spherical wave phase. Then, they are propagated to the pupil plane and multiplied by the corresponding aperture filter function. The size of the filter aperture is variable which is determined by the depth of the object. Finally, the complex amplitudes at the pupil plane are back-propagated to the holographic plane and added together to form the DOF-enhanced hologram. Simulation and experimental results verify the proposed method could improve the DOF of holographic SMV display, which will contribute to the application of 3D NED.

Effect of Insoluble Dietary Fiber Extracted from Feijoa (Acca sellowiana (O. Berg) Burret.) Supplementation on Physicochemical and Functional Properties of Wheat Bread.

This study aimed to assess the effects of insoluble dietary fiber (IDF) from feijoa supplementation on the physicochemical and functional properties of wheat bread. The results showed that feijoa IDF (FJI) had the typical structures of hydrolysis fiber, polysaccharide functional groups, and crystal structure of cellulose. The gradual increase of FJI levels (from 2 to 8%) in wheat bread resulted in increased total DF, ash, and protein contents, accompanied by a reduction in moisture, carbohydrates, and energy value. The inclusion of FJI in the bread crumb caused a rise in both redness (a*) and yellowness (b*) values while decreasing the brightness (L*) relative to the control specimen. In addition, adding FJI up to 2% significantly increased total phenolic and flavonoid contents and antioxidant activity, as well as flavor score of supplemented bread samples, while additions above 2% resulted in undesirable taste and texture. FJI addition caused higher bile acid, NO2-, and cholesterol adsorption capacities. Moreover, FJI addition up to 4% significantly reduced glucose adsorption capacities at different in vitro starch digestion intervals. The findings revealed that FJI offers great potential as an ideal functional ingredient in food processing.

Comparison of quantitative volumetric analysis and linear measurement for predicting the survival of Barcelona Clinic Liver Cancer 0- and A stage hepatocellular carcinoma after radiofrequency ablation.

PURPOSE: The prognostic role of the tumor volume in patients with hepatocellular carcinoma (HCC) at the Barcelona Clinic Liver Cancer (BCLC) 0 and A stages remains unclear. This study aims to compare the volumetric measurement with linear measurement in early HCC burden profile and clarify the optimal cut-off value of the tumor volume. METHODS: The consecutive patients diagnosed with HCC who underwent initial and curative-intent radiofrequency ablation (RFA) were included retrospectively. The segmentation was performed semi-automatically, and enhanced tumor volume (ETV) as well as total tumor volume (TTV) were obtained. The patients were categorized into high- and low-tumor burden groups according to various cutoff values derived from commonly used diameter values, X-tile software, and decision-tree analysis. The inter- and intra-reviewer agreements were measured using the intra-class correlation coefficient. Univariate and multivariate time-to-event Cox regression analyses were performed to identify the prognostic factors of overall survival. RESULTS: A total of 73 patients with 81 lesions were analyzed in the whole cohort with a median follow-up of 31.0 (interquartile range: 16.0­36.3). In tumor segmentation, excellent consistency was observed in intra- and inter-reviewer assessments. There was a strong correlation between diameter-derived spherical volume and ETV as well as ETV and TTV. As opposed to all linear candidates and 4,188 mm3 (sphere equivalent to 2 cm in diameter), ETV >14,137 mm3 (sphere equivalent to 3 cm in diameter) or 23,000 mm3 (sphere equivalent to 3.5 cm in diameter) was identified as an independent risk factor of survival. Considering the value of hazard ratio and convenience to use, when ETV was at 23,000 mm3, it was regarded as the optimal volumetric cut-off value in differentiating survival risk. CONCLUSION: The volumetric measurement outperforms linear measurement on tumor burden evaluation for survival stratification in patients at BCLC 0 and A stages HCC after RFA.