Rab25 is involved in hypospadias via the ß1 integrin/EGFR pathway.

BACKGROUND: Hypospadias is a common congenital abnormality of the penile. Abnormal regulation of critical genes involved in urethral development leads to hypospadias. We used the Rab25-/- mice and foreskin fibroblasts transfected with lentivirus in vitro and in vivo to investigate the role of Rab25 in hypospadias. METHODS: The expression levels of various molecules in tissue samples and foreskin fibroblasts were confirmed using molecular biology methods (western blotting, PCR, immunohistochemistry, etc.). A scanning electron microscope (SEM) was used to visualize the external morphology of genital tubercles (GTs) of gestation day (GD) 18.5 male wild-type (WT) and Rab25-/- mice. RESULTS: An expanded distal cleft and V-shaped urethral opening were observed in GD 18.5 Rab25-/- mice. We demonstrated that Rab25 mediated hypospadias through the ß1 integrin/EGFR pathway. In addition, silencing Rab25 inhibited cell proliferation and migration and promoted apoptosis in the foreskin fibroblasts; Ki-67- and TUNEL-positive cells were mainly concentrated near the urethral seam. CONCLUSION: These findings suggest that Rab25 plays an essential role in hypospadias by activation of ß1 integrin/EGFR pathway, and Rab25 is a critical mediator of urethral seam formation in GD18.5 male fetal mice.

Efficacy and Safety of High-Dose Vitamin D Supplementation vs Solifenacin or Standard Urotherapy for Overactive Bladder Dry in Children: A Randomized Clinical Trial.

PURPOSE: We investigated the efficacy and safety of high-dose vitamin D supplementation (VDS) plus standard urotherapy (SU) in managing overactive bladder dry in children. MATERIALS AND METHODS: A 3-arm, randomized clinical trial was performed at an academic center in China between January 2023 and June 2023. Eligible patients (n=303) were randomized to receive 8 weeks of high-dose VDS (vitamin D3 drops encapsulated as soft capsules, 2400 IU/d) plus SU (VDS + SU group; n=100), solifenacin (5-10 mg/d) plus SU (SOL + SU group; n=102), or SU alone (SU group; n=101). Reduction in voiding frequency was the primary outcome. Secondary outcomes encompassed improvement in urgency, nocturia, quality of life score, pediatric lower urinary tract symptom score, and participant satisfaction. Treatment-emergent adverse events were recorded within each group. RESULTS: Participants had a median age of 82.0 months and their baseline mean vitamin D level was 22.64 ng/mL. The VDS + SU group showed greater improvements in voids/d than the SOL + SU group (median difference 3.0; 95% CI, 2.0 to 3.5; P < .001) and the SU group (median difference 4.0; 95% CI, 3.0 to 5.0; P < .001) after intervention. The VDS + SU group also showed the greatest improvement in quality of life and pediatric lower urinary tract symptom scores. Patient satisfaction was similar between the SOL + SU and SU groups. The VDS + SU group did not exhibit a heightened risk of treatment-emergent adverse events compared to the other groups. CONCLUSIONS: High-dose VDS plus SU was effective and well-tolerated in managing overactive bladder dry in children, suggesting its potential as a novel therapeutic option for this population.

Risk factors and timing of complication presentation following primary hypospadias repair in adolescents.

OBJECTIVES: To evaluate the risk factors for postoperative complications in adolescents who undergo primary hypospadias repair and determine the time required for complication detection. METHODS: Our study included patients classified as Tanner stages three to five who underwent primary hypospadias repairs at our hospital from January 2015 to August 2022. The patients' baseline information, clinical characteristics, postoperative complications, and time to complication detection were collected. Cox regression analysis, ROC curves, Kaplan-Meier survival analyses, and the Mann-Whitney U test were used. RESULTS: The study comprised 143 patients, with a median age of 12.58 years. Postoperative complications were experienced by 66 patients. The length of the urethral defect was identified as an independent risk factor for postoperative complications. The ROC curve analysis identified 3 cm as the optimal cutoff value for the length of the urethral defect. The median time to complication detection was 30.5 days (IQR 23 to 209.25). 89.4% of the complications were identified within the first year. Patients with a urethral defect of <3 cm experienced a significantly longer time for the detection of urethral fistula compared to those with a urethral defect of ≥3 cm (p = 0.047). CONCLUSIONS: Our data indicate that adolescents with a urethral defect ≥3 cm have a higher risk of postoperative complications. Although most complications were identified within the first year, conducting long-term follow-ups for adolescents is recommended to identify potential subsequent complications that may arise from persistent urethral alterations.

A grading system for evaluation of bladder trabeculation.

PURPOSE: To establish a parameter-based grading system for evaluating bladder trabeculation (BT). MATERIALS AND METHODS: A retrospective analysis was conducted on children diagnosed with posterior urethral valve (PUV) or neurogenic bladder (NB) who underwent voiding cystourethrogram (VCUG), urodynamic testing, and urological ultrasonography between January 2016 and October 2022. Cases involving urologic surgery, secondary bladder pathology, and an interval of more than 12 months between examinations were excluded. A parameter named Bladder Dispersion (BD) was calculated through fluoroscopic images, and the grading system was developed as follows: BD < 40 (Grade 0), 40 ≤ BD < 60 (Grade 1), 60 ≤ BD < 90 (Grade 2), BD ≥ 90 (Grade 3). Grades 0-1 were classified as low-risk group, while grades 2-3 were classified as high-risk group. Analysis of variance, Kruskal-Wallis test, and Chi-square test were performed to compare urodynamic results and complications across different grades and groups. RESULTS: A total of 74 patients were eligible to participate, which included 46 boys (62.2%) and 28 girls (37.8%), the mean age was 75.18 ± 48.39 months. Among them, 11 (14.9%) were PUV, 50 (67.6%) were NB, and 13 (17.5%) were PUV and NB. Significant differences were observed in maximum detrusor pressure, post-void residual urine ratio, and compliance among grades 0-3. Severe hydronephrosis and histories of urinary tract infection were more prevalent in the high-risk group. CONCLUSION: A reliable grading system with objective standards was proposed which could aid in the assessment of BT severity.

Design, synthesis and evaluation of quinoline-O-carbamate derivatives as multifunctional agents for the treatment of Alzheimer's disease.

A series of novel quinoline-O-carbamate derivatives was rationally designed for treating Alzheimer's disease (AD) by multi-target-directed ligands (MTDLs) strategy. The target compounds were synthesised and evaluated by AChE/BuChE inhibition and anti-inflammatory property. The in vitro activities showed that compound 3f was a reversible dual eeAChE/eqBuChE inhibitor with IC50 values of 1.3 µM and 0.81 µM, respectively. Moreover, compound 3f displayed good anti-inflammatory property by decreasing the production of IL-6, IL-1ß and NO. In addition, compound 3f presented significant neuroprotective effect on Aß25-35-induced PC12 cell injury. Furthermore, compound 3f presented good stabilities in artificial gastrointestinal fluids, liver microsomes in vitro and plasma. Furthermore, compound 3f could improve AlCl3-induced zebrafish AD model by increasing the level of ACh. Therefore, compound 3f was a promising multifunctional agent for the treatment of AD.

Rapid response of midgut bacteria in Bactrocera tau (Walker) (Diptera: Tephritidae) to lambda-cyhalothrin- and spinosad-induced stress.

In recent decades, the increasingly widespread application of chemical pesticides has exacerbated the emergence of insecticide resistance among insect pests. In this study, we examined the rapid response of bacteria in the midgut of the fruit fly Bactrocera tau (Walker) (Diptera: Tephritidae) to stress induced by the insecticides lambda-cyhalothrin and spinosad by analyzing the bacterial community structure and diversity in the midguts of 4-day-old B. tau. The results revealed that 4-day-old B. tau females were more resistant to lambda-cyhalothrin and spinosad than their 4-day-old male counterparts. Alpha- and beta-diversity analyses revealed no significant differences between male and female B. tau with respect to the diversity and richness of gut bacteria in response to the same treatments. In response to treatment with lambda-cyhalothrin and spinosad at lethal concentration 50 (LC50), we detected significant changes in the structure and diversity of the bacterial community in the midguts of both male and female B. tau. Particularly among the dominant bacterial genera, there were decreases in the relative abundances of Citrobacter, Enterobacter, Klebsiella, and Pectobacterium. Increases were observed in the relative abundances of Dysgonomonas, Erwinia, and Providencia. Our findings provide a theoretical basis for gaining a better understanding of the relationships between midgut bacteria and the insecticide resistance of B. tau.

Microclimate stability on the critical zone of a karst hillslope in southwest China: Insights from continuous temperature observations at the air-soil-epikarst interface.

Temperature is an important near-surface microclimate parameter that plays a key role in hydrological, ecological, and biogeochemical functions. However, the spatio-temporal distribution of temperature on the invisible and inaccessible soil-weathered bedrock continuum, wherein hydrothermal processes are most active, remains poorly understood. Temperature dynamics were monitored at 5 min intervals in the air-soil-epikarst (∼3 m) system at different topographical positions of the karst peak-cluster depression in southwest China. The weathering intensity was characterized based on the physicochemical properties of samples collected through drilling. No significant difference was observed in air temperature across slope positions, which was related to the limited distance and elevation resulting in roughly consistent energy input. The control effect of air temperature on the soil-epikarst was weakened with the decrease in elevation (±0.36 to ±0.25 °C). It is attributed to the enhanced temperature regulation capacity of vegetation cover from the up slope (shrub dominant) to down slope (tree dominant) in a relatively uniform energy environment. Temperature stability is clearly distinguished in two adjacent hillslopes that were differentiated by weathering intensity. For every 1 °C change in the ambient temperature, the amplitude of soil-epikasrt temperature variation on the strongly and weakly weathered hillslopes were ±0.28 and ± 0.32 °C, respectively. The response of soil-epikarst temperature to ambient temperature was more sensitive in the wet season (±0.40 °C) than in the dry season (±0.20 °C), which was related to the cooling effect caused by abundant rainfall. The cooling effect was particularly prominent in the preferential flow development area composed of pipeline cracks, which appear in the hillslope with relatively weak weathering intensity. These demonstrate that soil-epikarst temperature responds more gently to the variability of rainfall and ambient temperature on a relatively strong weathered hillslope. Accordingly, this study highlights that the sensitivity of soil-epikarst temperature to climate change is regulated by vegetation and weathering intensity on karst hillslopes in southwest China.

Computational fluid dynamics modeling approaches to assess lower urinary tract hydraulic dynamics in posterior urethral valve before and after endoscopic valve ablation: a pilot study.

PURPOSE: Computational fluid dynamics (CFD) has been used successfully in cardiovascular system research to analyze the physiological processes inside vessels. We evaluated the hydraulic information of urine through the lower urinary tract in a patient with posterior urethral valve (PUV) before and after valve ablation by CFD. METHODS: A set of models of the lower urinary tract were developed based on geometrical data obtained by cystoscopy and voiding cystourethrography. Simulated assumptions and conditions were applied according to prior studies and urodynamic results. We used Fluent CFD 19.0 (Ansys Inc., USA) to compute the velocity and pressure of the fluid regions. The simplification of Bernoulli's formula was applied afterward to calculate the hydraulic energy of different positions. RESULTS: The urine flow rates of the NORMALst, the PUVst, and the POSTst at 5000 Pa were 18.08 ml/s, 11.14 ml/s, and 12.16 ml/s, respectively. Precipitous pressure change was observed around the valve in the PUVst, and the abnormal change was concentrated in the dilated urethra in the POSTst. Major energy dissipations were generated around the valve and the dilated urethra in the PUVst. The energy loss that occurred in the dilated urethra did not improve after the operation. CONCLUSIONS: Our findings are probably indicative of the hydrodynamics changes in the dilated urethra in PUV and need to be confirmed through more improved CFD models in the future. CFD may revolutionize pediatric urologists' perception in the management of urinary disease.

Synergistic effects of nano-structured WO3-Se heterojunction decorated by organic Nafion layer on improving photoelectrochemical performance.

Exploration of high-performance photoanodes is considered as an essential challenge in photoelectrochemical (PEC) water splitting due to the complex four-electron reaction in water oxidation. Herein, the nano-structured WO3-Se heterojunction decorated by organic Nafion layer is designed. The optimized WO3-Se200-0.05Nafion photoanode shows a remarkable photocurrent of 1.40 mA cm-2at 1.23 V versus reversible hydrogen electrode, which is 2.5-fold higher than that of pure WO3nanosheets (WO3NS) photoelectrode. Remarkably, the photocurrent increments of WO3-Se200-0.05Nafion is larger than the increment sum of WO3-Se200 and WO3-0.05Nafion, which affirming the synergistic effect of Se nanospheres and Nafion layer. The improved PEC performances are attributed to the quick charge separation and transfer, the increased electric conductivity, and the excellent kinetics of oxygen evolution, which is derived from the strong interaction among WO3, Se and Nafion. Meanwhile, the better visible-light harvesting from Se nanospheres as photosensitizer and the induction of transparent Nafion as a passivation layer can explain this synergy. It hopes this heterostructure design with organic Nafion decoration can inspire to exploit outstanding performance photoanodes for PEC water splitting.

Effects of ferulic acid on muscle development and intestinal microbiota of zebrafish.

Ferulic acid (FA) is one of a common ingredients in Chinese herbal medicine. FA has the interesting property of promoting growth and improving meat quality in livestock, but the mechanism is not understood. This study evaluated both safety and mechanism of efficacy in zebrafish model. At 15 µg/mL or above, FA led to pericardial oedema and delayed growth in zebrafish embryos. Dietary FA promoted growth and feed assimilation in male adult zebrafish. Genes related to myogenic development (myod1, myog and myf5) were significantly upregulated by FA and muscle fibre width in skeletal muscle was increased. At 20 µg/g, FA significantly increased number of goblet cells in zebrafish intestinal tissue, and gut microbiota composition also changed. Based on 16s rRNA gene sequences, 20 µg/g FA decreased Firmicutes and increased Bacteroides. 20 µg/g FA also stimulated the expression of PPAR-α, a gene associated with fat metabolism, and decreased the expression of PPAR-ß and PPAR-γ. These gene expression changes were beneficial to fatty acid synthesis and metabolism and decreased fat deposition. Our overall results indicated that FA can be a safe growth promotor in fish particularly in skeletal muscles.

[Expert consensus on measurement and clinical application of serum HBV RNA in patients with chronic HBV infection].

Since the discovery of circulating hepatitis B virus (HBV) RNA in the peripheral blood of patients with chronic hepatitis B in 1996, a growing number of studies have focused on clarifying the biological characteristics and clinical application value of serum HBV RNA. This consensus mainly summarizes the research progress of serum HBV RNA existing profiles, quantitative detection methods, and current clinical applications. In order to better apply this indicator for the clinical management of patients with chronic HBV infection, recommendations on quantitative detection target regions, detection results, and clinical applications are put forward.

The Chinese Society of Hepatology position statement on the redefinition of fatty liver disease.

Fatty liver disease associated with metabolic dysfunction is of increasing concern in mainland China, the world's most populous country. The incidence of fatty liver disease is highest in China, surpassing the incidence in European countries and the USA. An international consensus panel recently published an influential report recommending a novel definition of fatty liver disease associated with metabolic dysfunction. This recommendation includes a switch in name from non-alcoholic fatty liver disease (NAFLD) to metabolic (dysfunction)-associated fatty liver disease (MAFLD) and adoption of a set of positive criteria for disease diagnosis that are independent of alcohol intake or other liver diseases. Given the unique importance of this proposal, the Chinese Society of Hepatology (CSH) invited leading hepatologists and gastroenterologists representing their respective provinces and cities to reach consensus on alternative definitions for fatty liver disease from a national perspective. The CSH endorses the proposed change from NAFLD to MAFLD (supported by 95.45% of participants). We expect that the new definition will result in substantial improvements in health care for patients and advance disease awareness, public health policy, and political, scientific and funding outcomes for MAFLD in China.

Water uptake depth is coordinated with leaf water potential, water-use efficiency and drought vulnerability in karst vegetation.

Root access to bedrock water storage or groundwater is an important trait allowing plant survival in seasonally dry environments. However, the degree of coordination between water uptake depth, leaf-level water-use efficiency (WUEi) and water potential in drought-prone plant communities is not well understood. We conducted a 135-d rainfall exclusion experiment in a subtropical karst ecosystem with thin skeletal soils to evaluate the responses of 11 co-occurring woody species of contrasting life forms and leaf habits to a severe drought during the wet growing season. Marked differences in xylem water isotopic composition during drought revealed distinct ecohydrological niche separation among species. The contrasting behaviour of leaf water potential in coexisting species during drought was largely explained by differences in root access to deeper, temporally stable water sources. Smaller-diameter species with shallower water uptake, more negative water potentials and lower WUEi showed extensive drought-induced canopy defoliation and/or mortality. By contrast, larger-diameter species with deeper water uptake, higher leaf-level WUEi and more isohydric behaviour survived drought with only moderate canopy defoliation. Severe water limitation imposes strong environmental filtering and/or selective pressures resulting in tight coordination between tree diameter, water uptake depth, iso/anisohydric behaviour, WUEi and drought vulnerability in karst plant communities.

Predictive potential of Nomogram based on GMWG for patients with hepatocellular carcinoma after radical resection.

BACKGROUND: Since it's a challenging task to precisely predict the prognosis of patients with hepatocellular carcinoma (HCC). We developed a nomogram based on a novel indicator GMWG [(Geometric Mean of gamma-glutamyltranspeptidase (GGT) and white blood cell (WBC)] and explored its potential in the prognosis for HCC patients. METHODS: The patients enrolled in this study were randomly assigned to training and validation cohorts. And we performed the Least Absolute Shrinkage and Selection Operator proportional hazards model (LASSO Cox) model with clinical characteristics, serum indexes, and novel GMWG. Multivariate analysis was performed to build a nomogram. The performance of the nomogram was evaluated by C-index, the area under the receiver operating characteristic curve (AUC), and the calibration curve. Kaplan-Meier curves showed discrimination of the nomogram. Clinical utility was assessed by decision curve analysis (DCA). The discrimination ability of the nomogram was determined by the net reclassification index (NRI). RESULTS: The geometric mean of GGT and white WBC count (GMWG), neutrophil to lymphocyte ratio (NLR), and tumor size were significantly associated with the overall survival (OS). The variables above were used to develop the nomogram. The indexes of nomogram were 0.70 and 071 in the training or validation cohort, respectively. AUC of 1-, 3- and 5-year OS showed satisfactory accuracy as well. The calibration curve showed agreement between the ideal and predicted values. Kaplan-Meier curves based on the overall survival (OS) and disease-free survival (DFS) showed significant differences between nomogram predictive low and high groups. DCA showed clinical utilities while NRI showed discrimination ability in both training or validation cohort. CONCLUSIONS: GMWG might be a potential prognostic indicator for patients with HCC. The nomogram containing GMWG also showed satisfaction prediction capacity.

Larger males facilitate population expansion in Ophraella communa.

One of the most intriguing concepts in animal ecology is the reproductive advantages offered by larger body size, and the females prefer to mate with larger males to gain reproductive advantage. Currently, it is not clear how females recognize signs of male 'quality' and what mechanisms are involved in producing offspring with direct or indirect benefits. Our study aims to assess the preferences of females for males in Ophraella communa, determine the reproductive benefits and reveal the underlying mechanism behind this advantage. We demonstrate that male body size is an important determinant in the evolutionary process of O. communa, affecting female mate choice. Moreover, our study establishes that females prefer males with a larger body size, and this could further improve the developmental and reproductive fitness of their offspring. Finally, we focus on the seminal fluid proteins (SFPs) in O. communa, determine differentially expressed genes (i.e. OcACE, OcCBP and OcSFP) by analysing their proteomes and transcriptomes, and define the role of these SFPs-related genes through RNAi. Our study proved that the reproductive benefit of large males may be regulated by biased expression of crucial SFPs genes. The present study advances our understanding of the biological significance of preferential mating.

The significance of m6A RNA methylation regulators in predicting the prognosis and clinical course of HBV-related hepatocellular carcinoma.

BACKGROUND: Hepatocarcinogenesis is reportedly correlated with abnormal m6A modifications; however, it is unknown whether m6A RNA methylation regulators facilitate the occurrence of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Thus, we constructed an m6A-related model that may enhance HBV-related HCC prognosis. METHODS: Gene signatures of HNRNPA2B1 and RBM15 were generated by univariate and Lasso Cox regression analyses using the gene set and clinical information from The Cancer Genome Atlas (TCGA) database. High-risk and low-risk groups were confirmed based on the gene signature model. Furthermore, we validated the predictive roles of the two genes for overall survival (OS) in the GSE14520 dataset. The relative expression of 22 paired mRNAs was measured using quantitative real-time polymerase chain reaction (qRT-PCR) analysis to determine whether the two genes had a predictive role in our Guilin cohort. RESULTS: The differences in OS between the high-risk and low-risk groups were statistically significant in the TCGA (p = 0.003) and GSE14520 (p = 0.045) datasets, but not in the Guilin cohort, owing to differences in clinical information among the three cohorts (mainly the TNM stage and survival state). Stratified analysis of TNM stages showed that the two-gene signature acted as a prognostic indicator of HBV-related HCC patients in the early TNM stage; both TCGA and GSE14520 cohorts showed statistical significance. Moreover, multivariate Cox regression analysis indicated that the two-gene signature was an independent factor for predicting prognosis (HR = 1.087, 95% CI: 1.007-1.172). Correlation analysis between the gene signature and clinical features revealed that the risk stratification was significantly correlated with grade and survival state. Finally, Gene Set Enrichment Analysis (GSEA) revealed that the KEGG pathways associated with the cell cycle, DNA replication, the spliceosome, repair, and metabolism-related processes were all significantly enriched in the high-risk group. Among the enriched genes, the expression levels of the replication protein RPA1 and the pre-mRNA splicing factor SF3B1 were significantly upregulated in the high-risk group. These results might help in elucidating the underlying molecular mechanisms of HBV-related HCC. CONCLUSIONS: Our data may provide new predictive signatures and potential therapeutic targets to identify and treat HBV-related HCC patients in the early disease stage.

Olfactory co-receptor is involved in host recognition and oviposition in Ophraella communa (Coleoptera: Chrysomelidae).

Common ragweed (Ambrosia artemisiifolia) is a notorious invasive weed that has spread across most temperate regions of the world. The beetle (Ophraella communa) is considered to be an effective control agent against A. artemisiifolia. As an oligophagous insect, its olfactory system is extremely important for host seeking in the wild. To the best of our knowledge, there is no report on the molecular mechanisms underlying olfaction recognition in this beetle. Hence, in this study, we characterized the odorant receptor co-receptor of O. communa and named it as 'OcomORco'. Real-time quantitative PCR (qRT-PCR) showed that, compared to the control treatment, RNA interference (RNAi) strongly reduced the expression of OcomORco by 89% in male and 90% in female beetles. Electroantennogram assay showed that the antennal response of both male and female beetles to four volatiles of A. artemisiifolia was significantly reduced. The injected male or female beetles lost their preference for plant leaves as observed in the behavioural tests. In addition, disruption of the expression of OcomORco resulted in a reduction of oviposition, while there was no difference in larval hatching rate between control and knockdown females. We demonstrated that OcomORco plays a vital role in olfactory perception and host search in O. communa, and it is involved in oviposition in an indirect way.

PPPDE1 promotes hepatocellular carcinoma development by negatively regulate p53 and apoptosis.

We have previously identified that PPPDE1 is a deubiquitinase (DUB) belonging to a cysteine isopeptidase family. Here we sought to explore the biological significance of PPPDE1 in hepatocellular carcinoma and its underlying molecular mechanism. In the present study, we found that amplification and overexpression of PPPDE1 were associated with poor prognosis in hepatocellular carcinoma (HCC). We also demonstrated that knocking down of PPPDE1 could significantly block the clonal growth and tumorigenicity of human HCC cells, which revealed a critical role for PPPDE1 in HCC development. Furthermore, we proved that PPPDE1 is a key modulator of p53 protein level and its down stream apoptosis pathway. Taken together, these results suggested that PPPDE1 is a putative HCC driver gene and extensive studies should be conducted in the future to investigate the role of PPPDE1 in HCC and other tumors.

A Large-scale, multicenter serum metabolite biomarker identification study for the early detection of hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is the third most lethal cancer worldwide. The lack of effective biomarkers for the early detection of HCC results in unsatisfactory curative treatments. Here, metabolite biomarkers were identified and validated for HCC diagnosis. A total of 1,448 subjects, including healthy controls and patients with chronic hepatitis B virus infection, liver cirrhosis, and HCC, were recruited from multiple centers in China. Liquid chromatography-mass spectrometry-based metabolomics methods were used to characterize the subjects' serum metabolic profiles and to screen and validate the HCC biomarkers. A serum metabolite biomarker panel including phenylalanyl-tryptophan and glycocholate was defined. This panel had a higher diagnostic performance than did α-fetoprotein (AFP) in differentiating HCC from a high-risk population of cirrhosis, such as an area under the receiver-operating characteristic curve of 0.930, 0.892, and 0.807 for the panel versus 0.657, 0.725, and 0.650 for AFP in the discovery set, test set, and cohort 1 of the validation set, respectively. In the nested case-control study, this panel had high sensitivity (range 80.0%-70.3%) to detect preclinical HCC, and its combination with AFP provided better risk prediction of preclinical HCC before clinical diagnosis. Besides, this panel showed a larger area under the receiver-operating characteristic curve than did AFP (0.866 versus 0.682) to distinguish small HCC, and 80.6% of the AFP false-negative patients with HCC were correctly diagnosed using this panel in the test set, which was corroborated by the validation set. The specificity and biological relevance of the identified biomarkers were further evaluated using sera from another two cancers and HCC tissue specimens, respectively. Conclusion: The discovered and validated serum metabolite biomarker panel exhibits good diagnostic performance for the early detection of HCC from at-risk populations. (Hepatology 2018;67:662-675).