Overview of mechanisms and novel therapies on rheumatoid arthritis from a cellular perspective.

Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovial inflammation of joints in response to autoimmune disorders. Once triggered, many factors were involved in the development of RA, including both cellular factors like osteoclasts, synovial fibroblasts, T cells, B cells, and soluble factors like interleukin-1 (IL-1), IL-6, IL-17 and tumor necrosis factor-α (TNF-α), etc. The complex interplay of those factors results in such pathological abnormality as synovial hyperplasia, bone injury and multi-joint inflammation. To treat this chronic life-affecting disease, the primary drugs used in easing the patient's symptoms are disease-modifying antirheumatic drugs (DMARDs). However, these traditional drugs could cause serious side effects, such as high blood pressure and stomach ulcers. Interestingly, recent discoveries on the pathogenesis of RA have led to various new kinds of drugs or therapeutic strategies. Therefore, we present a timely review of the latest development in this field, focusing on the cellular aspects of RA pathogenesis and new therapeutic methods in clinical application. Hopefully it can provide translational guide to the pre-clinical research and treatment for the autoimmune joint disease.

Safety of flexible ureteroscopic lithotripsy with 365 µm fibers of Ho: YAG laser: a short-term follow-up.

Our previous study first reported the efficacy of FURL using 365 µm fibers with holmium: yttrium aluminum garnet (Ho: YAG) laser lithotripsy. This study evaluates the 16-week safety of this procedure. A prospective cohort study was conducted, and the clinical characteristics of patients who underwent FURL using 365 µm fibers with Ho: YAG laser were retrospectively collected. Descriptive statistics were reported, and logistic regression analysis was conducted to identify risk factors influencing the stone-free rate (SFR). Infection-related laboratory parameters, including white blood cell count (WBC), procalcitonin (PCT), and C-reactive protein (CRP), were collected. Regression analysis was conducted to identify risk factors for the development of urosepsis post-surgery. Additionally, a 16-week follow-up was conducted in outpatient clinics, and kidney function was assessed. A total of 274 patients participated in this study. The 4-week stone-free rate (SFR) following FURL with 365 µm fibers of Ho: YAG laser was significantly associated with stone size and composition. No severe complications were observed following FURL procedures. There were no significant differences in white blood cell count (WBC), C-reactive protein (CRP), and procalcitonin (PCT) in peripheral blood before and after FURL procedures. Urosepsis was diagnosed in 1.82% of patients. Preoperative white blood cell count (WBC) in urine and preoperative urine culture results were identified as significant risk factors. Kidney function remained stable at 4 and 16 weeks following FURL. This prospective cohort study demonstrated the high safety of FURL with 365 µm fibers of Ho: YAG laser, as infection-related parameters during the perioperative period showed no significant differences, and kidney function remained stable throughout the 16-week follow-up.

Clinical characteristics and factors associated with mucus plugs under bronchoscopy in children hospitalized for acute asthma attack.

Objective: To describe clinical characteristics of hospitalized children with acute asthma attacks complicated with mucus plugs and to investigate the factors associated with mucus plugs in asthma children. Methods: This retrospective study analyzed hospitalized children and adolescents with acute asthma attacks from January 2016 to December 2021. The demographic information and characteristics were collected. Subjects were categorized into the mucus plug group and the control group based on the bronchoalveolar lavage results. The Logistic regression analyses were utilized to assess the relative factors associated with mucus plugs. All data were analyzed using SPSS 22.0. Results: This study included 242 individuals. Out of the 151 subjects who underwent bronchoscopy, 62.9% were classified in the mucus plug group and 37.1% in the control group. The subjects with dyspnea had a higher proportion in the mucus plug group (52.6% vs. 26.8%). The serum total IgE level of the mucus plug group was lower than the control group. The proportion of subjects who were diagnosed with asthma for the first time during hospitalization (87.4% vs. 76.8%) and combined with respiratory infection (91.6% vs. 82.1%) in the mucus plug group might be higher than that in the control group. More subjects in the mucus plug group were administered systemic glucocorticoid, magnesium sulfate, aminophylline, and mucolytic drugs after hospitalization. In multivariable analysis, diagnosed with asthma for the first time during hospitalization (OR = 4.404; 1.101-17.614), dyspnea (OR = 4.039; 1.306-12.496), and cesarean (OR = 0.274; 0.092-0.812) might be associated with mucus plug in children hospitalized for an acute asthma attack. Suggests: While our retrospective study suggests that some clinical features of children hospitalized with asthma who have mucus plugs differ from those without, further studies are required.

Enhanced Photocatalytic Hydrogen Production Activity Driven by TiO2/(MoP/CdS): Insights from Powder Particles to Thin Films.

Transitioning from powder photocatalysts to thin film photocatalysts is one of the necessary steps toward industrializing photocatalytic hydrogen production. Herein, we reported the integration of non-noble metal cocatalyst MoP decorated with TiO2 and CdS, forming TiO2/(MoP/CdS) for ultraviolet-visible light utilization. The designed powder TiO2/(MoP/CdS) composites achieved a superior hydrogen production rate of 42.2 mmol g-1 h-1, which is 30.1 times that of TiO2/CdS, performing the highest activity among the TiO2-CdS-based composite photocatalysts. Moreover, we fabricated a thin film from TiO2/(MoP/CdS) powder, which exhibited comparable photocatalytic activity for hydrogen production, achieving 35.5 mmol g-1 h-1 and maintaining long-term stability for 150 h. The outstanding performance was attributed to the ability of the TiO2/(MoP/CdS) composite photocatalysts to absorb both visible and ultraviolet light. Additionally, the heterojunction formed between TiO2 and CdS also played a significant role in the overall photocatalyst activity. This cost-effective catalyst holds promise for future large-scale industrial applications.

Insight into Grain Refinement Mechanisms of WC Cemented Carbide with Al0.5CoCrFeNiTi0.5 Binder.

High-entropy alloys (HEA) as a kind of new binder for cemented carbide have garnered significant attention. In this work, WC/(17~25 wt.%)Al0.5CoCrFeNiTi0.5 cemented carbides were prepared by hot pressing sintering (HPS), and the reactions between WC powder and Al0.5CoCrFeNiTi0.5 powder during hot pressing sintering were elucidated. It found that different from traditional Co binder, the Al0.5CoCrFeNiTi0.5 binder effectively inhibited WC grain growth. During HPS, the decomposed W and C atoms from WC diffused into the Al0.5CoCrFeNiTi0.5 binder, reacted with the elements in the binder, and then formed the M(Co, Fe, Ni)3W3C phase. The back-diffusion of W and C atoms to WC grains was restricted by the Al0.5CoCrFeNiTi0.5 alloy and inhibited them from re-precipitating onto the large undissolved WC grains. As a result, the average size of WC grains in the cemented carbides was less than 200 nm. This work bright new insight into the grain refinement mechanisms of WC cemented carbide with HEA binder and provide a guidance for designing performance-stable WC/HEA cemented carbide and promoting their application.

Electrical Double Layer and In Situ Polymerization SEI Enables High Reversible Zinc Metal Anode.

Aqueous zinc-ion batteries (AZIBs) stand out among new energy storage devices due to their excellent safety and environmental friendliness. However, the formation of dendrites and side reactions on the zinc metal anode during cycling have become the major obstacles to their commercialization. This study innovatively selected Sodium 4-vinylbenzenesulfonate (VBS) as a multifunctional electrolyte additive to address the issues. The dissociated VBS- anions can not only significantly alter the hydrogen bond network structure of H2O in the electrolyte, but also preferentially adsorb on the surface of the zinc anode before H2O molecules, which will result in the development of organic anion-rich interface and alterations to the electrical double layer (EDL) structure. Furthermore, the ─C═C─ structure in VBS leads to the formation of an in situ polymerized organic anion solid electrolyte interface (SEI) layer that adheres to the surface of the zinc anode. The mechanisms work together to significantly improve the performance of Zn//Zn symmetric batteries, achieving a cycle life of over 1800 h at 1 mA cm-2 and 1 mAh cm-2. The introduction of VBS also enhances the cycling performance and capacity of Zn//δ-MnO2 full cells. This study provides a low-cost solution for the development of AZIBs.

[Establishment and optimization of Agrobacterium tumefaciens-mediated genetic transformation system for Polyporus umbellatus].

The unstable quality of Polyporus umbellatus sclerotia during cultivation is the key factor affecting the quality and yield of P. umbellatus sclerotia. In order to provide technical support for obtaining superior P. umbellatus by molecular breeding, the genetic transformation system mediated by Agrobacterium tumefaciens was studied in this paper. A. tumefaciens-mediated method was used to investigate the effects of antibiotic concentration, strain type, A. tumefaciens concentration, receptor material, infection time, co-culture time, and screening conditions on the genetic transformation efficiency of P. umbellatus. The transformants were screened and detected by hygromycin resistance marker genes, polymerase chain reaction(PCR) of specific primers, and fluorescence detection methods. The results showed that the A. tumefaciens GV3101 strain could genetically transfer P. umbellatus mycelium cells, and the optimal conditions for infection were as follows: the A. tumefaciens concentration A_(600 nm)= 0.6, P. umbellatus mycelium cells as receptor material, infection time of 30 min, and co-culture time of 3 days. The two-step screening method involving hygromycin of 9 and 13 µg·mL~(-1 )was the best screening condition. The results of hygromycin resistance screening, PCR detection of specific primers, and fluorescence detection showed that the exogenous gene eGFP had been transferred into the P. umbellatus mycelium cells, integrated into the genome, and successfully expressed. Under optimal conditions, the conversion efficiency could be increased to 2.3%, and the genetic transformation period was shortened from more than 90 days to less than 60 days. This study established and optimized the genetic transformation system of P. umbellatus mycelium cells mediated by A. tumefaciens, laying a foundation for the analysis of the molecular mechanism of P. umbellatus during growth and molecular breeding.

Repurposing existing drugs for the treatment ofCOVID-19/SARS-CoV-2: A review of pharmacological effects and mechanism of action.

Following the coronavirus disease-2019 outbreak caused by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), there is an ongoing need to seek drugs that target COVID-19. First off, novel drugs have a long development cycle, high investment cost, and are high risk. Second, novel drugs must be evaluated for activity, efficacy, safety, and metabolic performance, contributing to the development cycle, investment cost, and risk. We searched the Cochrane COVID-19 Study Register (including PubMed, Embase, CENTRAL, ClinicalTrials.gov, WHO ICTRP, and medRxiv), Web of Science (Science Citation Index, Emerging Citation Index), and WHO COVID-19 Coronaviral Disease Global Literature to identify completed and ongoing studies as of February 20, 2024. We evaluated the pharmacological effects, in vivo and in vitro data of the 16 candidates in the paper. The difficulty of studying these candidates in clinical trials involving COVID-19 patients, dosage of repurposed drugs, etc. is discussed in detail. Ultimately, Metformin is more suitable for prophylactic administration or mildly ill patients; the combination of Oseltamivir, Tamoxifen, and Dexamethasone is suitable for moderately and severely ill patients; and more clinical trials are needed for Azvudine, Ribavirin, Colchicine, and Cepharanthine to demonstrate efficacy.

An unprotected vulnerable relict subtropical conifer-Keteleeria evelyniana: Its forests, populations, growth and endangerment by invasive alien plant species in China.

Relict subtropical coniferous forests in China face severe fragmentation, resulting in declining populations, and some are under significant threat from invasive alien species. Despite the crucial importance of understanding forest dynamics, knowledge gaps persist, particularly regarding the impact of invasive plants on vulnerable natives like Keteleeria evelyniana. In this study, we investigated the impact of invasive plants on the regeneration of forests dominated by K. evelyniana, a subtropical relict species in southwestern China. For this purpose, we characterized forest dynamics of 160 forest plots featuring K. evelyniana as the primary dominant species and determined whether the presence of invasive plants was correlated with regeneration of K. evelyniana. We identified four distinct forest types in which K. evelyniana was dominant. We found that radial growth of K. evelyniana trees is faster in younger age-classes today than it was for older trees at the same age. The population structure of K. evelyniana in each forest type exhibited a multimodal age-class distribution. However, three forest types lacked established saplings younger than 10 years old, a situation attributed to the dense coverage of the invasive alien Ageratina adenophora. This invasive species resulted in a reduction of understory species diversity. Additionally, our analysis uncovered a significant negative correlation in phylogenetic relatedness (net relatedness index) between native and invasive alien plant species in eastern Yunnan. This suggests closely related invasive species face heightened competition, hindering successful invasion. Taken together, our findings indicate that successful establishment and habitat restoration of K. evelyniana seedling/saplings require effective measures to control invasive plants.

Extrachromosomal circular DNA orchestrates genome heterogeneity in urothelial bladder carcinoma.

Rationale: Extrachromosomal circular DNA is a hallmark of cancer, but its role in shaping the genome heterogeneity of urothelial bladder carcinoma (UBC) remains poorly understood. Here, we comprehensively analyzed the features of extrachromosomal circular DNA in 80 UBC patients. Methods: We performed whole-genome/exome sequencing (WGS/WES), Circle-Seq, single-molecule real-time (SMRT) long-read sequencing of circular DNA, and RNA sequencing (RNA-Seq) on 80 pairs of tumor and AT samples. We used our newly developed circular DNA analysis software, Circle-Map++ to detect small extrachromosomal circular DNA from Circle-Seq data. Results: We observed a high load and significant heterogeneity of extrachromosomal circular DNAs in UBC, including numerous single-locus and complex chimeric circular DNAs originating from different chromosomes. This includes highly chimeric circular DNAs carrying seven oncogenes and circles from nine chromosomes. We also found that large tumor-specific extrachromosomal circular DNAs could influence genome-wide gene expression, and are detectable in time-matched urinary sediments. Additionally, we found that the extrachromosomal circular DNA correlates with hypermutation, copy number variation, oncogene amplification, and clinical outcome. Conclusions: Overall, our study provides a comprehensive extrachromosomal circular DNA map of UBC, along with valuable data resources and bioinformatics tools for future cancer and extrachromosomal circular DNA research.

Genome-wide characterization of extrachromosomal circular DNA in SLE and functional analysis reveal their association with apoptosis.

Extrachromosomal circular DNA (eccDNA) derived from linear chromosomes, are showed typical nucleosomal ladder pattern in agarose gel which as a known feature of apoptosis and demonstrated to be immunogenicity. In systemic lupus erythematosus (SLE) patients, elevated levels of cell-free DNA (cfDNA) can be found in either linear forms or circular forms, while circular ones are much less common and harder to detect. The molecular characteristics and function of circular forms in plasma SLE patients remains elusive. Herein, we characterized the hallmarks of plasma eccDNA in SLE patients, including the lower normalized number and GC content of eccDNA in SLE plasma than in the healthy, and SLE eccDNA number positively correlated with C3 and negatively with anti-dsDNA antibodies. The differential eccGenes (eccDNAs carrying the protein coding gene sequence) of SLE was significantly enriched in apoptosis-related pathways. The artificially synthesized eccDNA with sequences of the PRF1 exon region could promote transcriptional expression of PRF1, IFNA and IFIT3 and inhibit early-stage apoptosis. Plasma eccDNA can serve as a novel autoantigen in the pathogenesis of SLE.

Multi-view heterogeneous graph learning with compressed hypergraph neural networks.

Multi-view learning is an emerging field of multi-modal fusion, which involves representing a single instance using multiple heterogeneous features to improve compatibility prediction. However, existing graph-based multi-view learning approaches are implemented on homogeneous assumptions and pairwise relationships, which may not adequately capture the complex interactions among real-world instances. In this paper, we design a compressed hypergraph neural network from the perspective of multi-view heterogeneous graph learning. This approach effectively captures rich multi-view heterogeneous semantic information, incorporating a hypergraph structure that simultaneously enables the exploration of higher-order correlations between samples in multi-view scenarios. Specifically, we introduce efficient hypergraph convolutional networks based on an explainable regularizer-centered optimization framework. Additionally, a low-rank approximation is adopted as hypergraphs to reformat the initial complex multi-view heterogeneous graph. Extensive experiments compared with several advanced node classification methods and multi-view classification methods have demonstrated the feasibility and effectiveness of the proposed method.

MiR-183-5p inhibits lung squamous cell carcinoma survival through disrupting hypoxia adaptation mediated by HIF-1α/NDUFA4L2 axis.

Hypoxia is a common feature of lung squamous cell carcinoma (LUSC), and hypoxia-inducible factor-1 (HIF-1) overexpression is associated with poor clinical outcome in LUSC. NADH dehydrogenase 1 alpha subcomplex subunit 4-like 2 (NDUFA4L2) is a recently identified target of HIF-1, but its roles in LUSC remain unclear. Herein, the expression and regulatory mechanisms of NDUFA4L2 were investigated in LUSC, and the influences on LUSC cell oxidative metabolism and survival of NDUFA4L2 were determined. The potential microRNA targeting to NDUFA4L2 was identified and its roles on LUSC cell were detected. We found that NDUFA4L2 were overexpressed in LUSC tissues, and that NDUFA4L2 expression correlated with shorter overall survival. NDUFA4L2 was regulated by HIF-1α under hypoxia, and NDUFA4L2 decreased mitochondrial reactive oxygen species (mitoROS) production through inhibiting mitochondrial complex I activity in LUSC cells. NDUFA4L2 silencing effectively suppressed LUSC cell growth and enhanced apoptosis by inducing mitoROS accumulation. Additionally, NDUFA4L2 was a target for miR-183-5p, and LUSC patients with high miR-183-5p levels had better prognoses. MiR-183-5p significantly induced mitoROS production and suppressed LUSC survival through negatively regulating NDUFA4L2 in vitro and in vivo. Our results suggested that regulation of NDUFA4L2 by HIF-1α is an important mechanism promoting LUSC progression under hypoxia. NDUFA4L2 inhibition using enforced miR-183-5p expression might be an effective strategy for LUSC treatment.

Herbivore-induced volatiles reduce the susceptibility of neighboring tomato plants to transmission of a whitefly-borne begomovirus.

Plant viruses exist in a broader ecological community that includes non-vector herbivores that can impact vector abundance, behavior, and virus transmission within shared host plants. However, little is known about the effects of non-vector herbivore infestation on virus transmission by vector insects on neighboring plants through inter-plant airborne chemicals. In this study, we investigated how volatiles emitted from tomato plants infested with the two-spotted spider mite (Tetranychus urticae) affect the infection of neighboring plants by tomato yellow leaf curl virus (TYLCV) transmitted by whitefly (Bemisia tabaci). Exposure of neighboring tomato plants to volatiles released from T. urticae-infested tomato plants reduced subsequent herbivory as well as TYLCV transmission and infection, and the jasmonic acid signaling pathway was essential for generation of the inter-plant defense signals. We also demonstrated that (E)-ß-ocimene and methyl salicylic acid were two volatiles induced by T. urticae that synergistically attenuated TYLCV transmission and infection in tomato. Thus, our findings suggest that plant-plant communication via volatiles likely represents a widespread defensive mechanism that substantially contributes to plant fitness. Understanding such phenomena may help us to predict the occurrence and epidemics of multiple herbivores and viruses in agroecosystems, and ultimately to manage pest and virus outbreaks.

Superelastic Polymer Aerogel with Superamphiphilicity.

Elastic aerogels have become a research hot spot in both academia and industry recently. The reported elastic aerogels are all made of hard materials by controlling their shapes. Herein we report an elastic aerogel made of a polymer elastomer with entropy elasticity. In the aerogel, cross-linked carboxyl nitrile rubber nanoparticles with hydrophilicity are dispersed in hydrophobic derivative of styrene-maleic anhydride alternating copolymer, forming a very special micro-nano surface structure with hydrophilic protrusions and hydrophobic depressions on the aerogel wall; therefore, the aerogel is not only superelastic but also superamphiphilic. A leak-free phase-change composite was prepared using the aerogel and paraffin, which can maintain at phase change temperature of paraffin for a longer time than the traditional one. The aerogel is also extremely suitable for desalination evaporators in solar-driven interfacial evaporation technology due to its superamphiphilicity, superelasticity, and ability to absorb sunlight. Exceptional evaporation rate of 2.78 kg·m-2·h-1 and evaporation efficiency of 170% could be reached even without using expensive light-absorbing materials. The evaporation rate exceeds that of most evaporators with expensive light-absorbing materials, and the evaporation efficiency exceeds the theoretical limit of conventional 2D solar evaporators. Both the phase-change composite and the evaporator can be easily recovered because the novel superelastic aerogel reported in this work is also recyclable.

Response Properties of Electrorheological Composite Hydrophilic Elastomers Based on Different Morphologies of Magnesium-Doped Strontium Titanate.

As smart materials, electrorheological elastomers (EREs) formed by pre-treating active electrorheological particles are attracting more and more attention. In this work, four Mg-doped strontium titanate (Mg-STO) particles with spherical, dendritic, flake-like, and pinecone-like morphologies were obtained via hydrothermal and low-temperature co-precipitation. XRD, SEM, Raman, and FT-IR were used to characterize these products. The results showed that Mg-STOs are about 1.5-2.0 µm in size, and their phase structures are dominated by cubic crystals. These Mg-STOs were dispersed in a hydrogel composite elastic medium. Then, Mg-STO/glycerol/gelatin electrorheological composite hydrophilic elastomers were obtained with or without an electric field. The electric field response properties of Mg-doped strontium titanate composite elastomers were investigated. We concluded that dendritic Mg-STO composite elastomers are high-performance EREs, and the maximum value of their energy storage was 8.70 MPa. The significant electrorheological performance of these products is helpful for their applications in vibration control, force transducers, smart structures, dampers, and other fields.

Decoding sunitinib resistance in ccRCC: Metabolic-reprogramming-induced ABAT and GABAergic system shifts.

Sunitinib, a primary treatment for clear cell renal cell carcinoma (ccRCC), frequently encounters the challenge of resistance development. Metabolic reprogramming, a characteristic change in ccRCC, is likely linked to this resistance. Our research revealed a notable decrease in the expression of the key metabolic gene ABAT in ccRCC, which contributed to diminished sensitivity to sunitinib. Downregulation of ABAT led to an increase in the intracellular level of gamma-aminobutyric acid (GABA), triggering abnormal activation of the G-protein-coupled receptor GABA-B. This activation resulted in increased transactivation of the tyrosine kinase receptors SYK and LYN, thereby reducing the antitumor and antiangiogenic properties of sunitinib. However, the application of SYK and LYN inhibitors successfully inhibited this effect. The transactivation of SYK and LYN caused resistance to the antiangiogenic effects of sunitinib through the upregulation of PGF protein levels. Furthermore, the combined application of an LYN inhibitor with sunitinib has been shown to enhance therapeutic efficacy.

Metabolic heterogeneity in tumor microenvironment - A novel landmark for immunotherapy.

The surrounding non-cancer cells and tumor cells that make up the tumor microenvironment (TME) have various metabolic rhythms. TME metabolic heterogeneity is influenced by the intricate network of metabolic control within and between cells. DNA, protein, transport, and microbial levels are important regulators of TME metabolic homeostasis. The effectiveness of immunotherapy is also closely correlated with alterations in TME metabolism. The response of a tumor patient to immunotherapy is influenced by a variety of variables, including intracellular metabolic reprogramming, metabolic interaction between cells, ecological changes within and between tumors, and general dietary preferences. Although immunotherapy and targeted therapy have made great strides, their use in the accurate identification and treatment of tumors still has several limitations. The function of TME metabolic heterogeneity in tumor immunotherapy is summarized in this article. It focuses on how metabolic heterogeneity develops and is regulated as a tumor progresses, the precise molecular mechanisms and potential clinical significance of imbalances in intracellular metabolic homeostasis and intercellular metabolic coupling and interaction, as well as the benefits and drawbacks of targeted metabolism used in conjunction with immunotherapy. This offers insightful knowledge and important implications for individualized tumor patient diagnosis and treatment plans in the future.

Prediction of antimicrobial resistance in Klebsiella pneumoniae using genomic and metagenomic next-generation sequencing data.

OBJECTIVES: Klebsiella pneumoniae is a significant pathogen with increasing resistance and high mortality rates. Conventional antibiotic susceptibility testing methods are time-consuming. Next-generation sequencing has shown promise for predicting antimicrobial resistance (AMR). This study aims to develop prediction models using whole-genome sequencing data and assess their feasibility with metagenomic next-generation sequencing data from clinical samples. METHODS: On the basis of 4170 K. pneumoniae genomes, the main genetic characteristics associated with AMR were identified using a LASSO regression model. Consequently, the prediction model was established, validated and optimized using clinical isolate read simulation sequences. To evaluate the efficacy of the model, clinical specimens were collected. RESULTS: Four predictive models for amikacin, ciprofloxacin, levofloxacin, and piperacillin/tazobactam, initially had positive predictive values (PPVs) of 92%, 98%, 99%, 94%, respectively, when they were originally constructed. When applied to clinical specimens, their PPVs were 96%, 96%, 95%, and 100%, respectively. Meanwhile, there were negative predictive values (NPVs) of 100% for ciprofloxacin and levofloxacin, and 'not applicable' (NA) for amikacin and piperacillin/tazobactam. Our method achieved antibacterial phenotype classification accuracy rates of 95.92% for amikacin, 96.15% for ciprofloxacin, 95.31% for levofloxacin and 100% for piperacillin/tazobactam. The sequence-based prediction antibiotic susceptibility testing (AST) reported results in an average time of 19.5 h, compared with the 67.9 h needed for culture-based AST, resulting in a significant reduction of 48.4 h. CONCLUSIONS: These preliminary results demonstrated that the performance of prediction model for a clinically significant antimicrobial-species pair was comparable to that of phenotypic methods, thereby encouraging the expansion of sequence-based susceptibility prediction and its clinical validation and application.