Diagnostic performance of angiography-derived fractional flow reserve and CT-derived fractional flow reserve: A systematic review and Bayesian network meta-analysis.

OBJECTIVE: Accumulating evidence has demonstrated that fractional flow reserves (FFRs) derived from invasive coronary angiograms (CA-FFRs) and coronary computed tomography angiography-derived FFRs (CT-FFRs) are promising alternatives to wire-based FFRs. However, it remains unclear which method has better diagnostic performance. This systematic review and meta-analysis aimed to compare the diagnostic performances of the two approaches. METHODS: The Cochrane Library, PubMed, Embase, Medline (Ovid), the Chinese China National Knowledge Infrastructure Database (CNKI), VIP, and WanFang Data databases were searched for relevant studies that included comparisons between CA-FFR and CT-FFR, from their respective database inceptions until January 1, 2023. Studies where both noninvasive FFR (including CA-FFR and CT-FFR) and invasive FFR (as a reference standard) were performed for the diagnosis of ischemic coronary artery disease and were designed as prospective, paired diagnostic studies, were pulled. The diagnostic test accuracy method and Bayesian hierarchical summary receiver operating characteristic (ROC) model for network meta-analysis (NMA) of diagnostic tests (HSROC-NMADT) were both used to perform a meta-analysis on the data. RESULTS: Twenty-six studies were included in this NMA. The results from both the diagnostic test accuracy and HSROC-NMADT methods revealed that the diagnostic accuracy of CA-FFR was higher than that of CT-FFR, in terms of sensitivity (Se; 0.86 vs. 0.84), specificity (Sp; 0.90 vs. 0.78), positive predictive value (PPV; 0.83 vs. 0.70), and negative predictive value (NPV; 0.91 vs. 0.89) for the detection of myocardial ischemia. A cumulative ranking curve analysis indicated that CA-FFR had a higher diagnostic accuracy than CT-FFR in the context of this study, with a higher area under the ROC curve (AUC; 0.94 vs. 0.87). CONCLUSIONS: Although both of these two commonly used virtual FFR methods showed high levels of diagnostic accuracy, we demonstrated that CA-FFR had a better Se, Sp, PPV, NPV, and AUC than CT-FFR. However, this study provided only indirect comparisions; therefore, larger studies are warranted to directly compare the diagnostic performances of these two approaches.

Bivalirudin versus heparin in contemporary percutaneous coronary interventions for patients with acute coronary syndrome: a systematic review and meta-analysis.

BACKGROUND: Bivalirudin is associated with fewer major bleeding events than heparin in patients undergoing percutaneous coronary intervention (PCI), but confounding effects of concomitant glycoprotein IIb/IIIa inhibitors, routine femoral artery access, and less potent effects of clopidogrel limits meaningful comparisons. The present study is a systematic review and meta-analysis to compare bivalirudin to heparin in contemporary practice. METHODS: The Cochrane Library, PubMed, EMBASE, and Ovid MEDLINE databases were searched for relevant studies, including comparisons between bivalirudin and heparin in the current medical era from inception to December 23, 2021. Studies reporting incidences of major adverse cardiac events (MACE) and net adverse clinical events (NACE) in patients undergoing PCI and meeting the inclusion criteria were retained. Data extraction was performed by three independent reviewers. RESULTS: The meta-analysis included 8 studies. Compared to heparin, bivalirudin during PCI was associated with a lower NACE risk, lower all-cause death, and similar MACE risk, with a pooled risk ratio of 0.82 (95% confidence interval [CI] 0.69-0.97, p = 0.02), 0.83 (95% CI 0.74-0.94, p = 0.002), and 0.93 (95% CI 0.78-1.10, p = 0.38), respectively. Moreover, the reduction in NACE was mainly attributed to reduced bleeding (22% reduction in the risk of major bleeding, 95% CI 0.63-0.97, p = 0.03). CONCLUSIONS: These findings suggest that bivalirudin use during PCI reduced the risk of NACE and all-cause death but did not reduce the risk of MACE compared with heparin use in PCI. More studies specifically designed for anticoagulation strategies and a personalized anticoagulation regimen to comprehensively balance bleeding and ischemia risks are required.

Rapid Kinetic Interactions of Sugar and Sugar Alcohol with Sweet Taste Receptors on Live Cells Using Stopped-Flow Spectroscopy.

The subjective measurement of the dynamic perception of sweetness is a problem in food science. Herein, the rapid interactions of sugars and sugar alcohols with sweet taste receptors on living cells on a millisecond timescale were studied via stopped-flow fluorescence spectroscopy. According to the rapid-kinetic parameters, sweeteners were divided into two groups. Sweeteners in group I disrupted the hydrogen bond network structure of water, and the apparent rate constant (kobs) was in the range of 0.45-0.6 s-1. Sweeteners in group II promoted the hydrogen bond formation of water, and the kobs was mostly in the range of 0.6-0.75 s-1. For most sweeteners, the kobs of cell responses was negatively correlated with the apparent specific volume of sweeteners. The differences in the cellular responses may be attributed to the disturbance in the water structure. Experimental results showed that the kinetic parameters of sweet cell responses reflected the dynamic perception of sweetness. Rapid kinetics, solution thermodynamic analysis, and water structure analysis enriched the physicochemical study of the sweetness mechanism and can be used to objectively evaluate the dynamic perception of sweetness.

Comparison of Different Chronic Maintenance Antithrombotic Strategies in Patients with Coronary Artery Disease: A Systematic Review and Network Meta-Analysis.

Background: Optimal antithrombotic therapy during the chronic maintenance period in patients with coronary artery disease (CAD) is unknown. We compared five kinds of mainstream chronic maintenance antithrombotic strategies at least one year after the acute phase: aspirin alone, clopidogrel alone, ticagrelor alone, continued dual antiplatelet therapy (DAPT) for a period of time, and maintenance with aspirin combined with a low-dose anticoagulant such as rivaroxaban. Methods: Ten randomized, controlled trials were selected using PubMed, Ovid MEDLINE, Embase, and Cochrane library through February 2023. The primary outcome was main adverse cardiac events (MACEs), and secondary outcomes include net adverse clinical events (NACEs), cardiac death, all-cause death, ischemic stroke, stent thrombosis, total bleeding, and major bleeding. A network meta-analysis was conducted with a random-effects model. Data extraction was performed by three independent reviewers. Results: Our search identified ten eligible randomized controlled trials enrolling a total of 82,084 patients comparing different chronic maintenance antithrombotic strategies. As for the primary endpoint, there was no statistical difference in MACE outcomes between any two of the five methods. As for the secondary endpoint, there was no statistical difference in NACE, major bleeding, all-cause death, cardiac death, and stent thrombosis between any two methods. The aspirin plus low-dose rivaroxaban group had a lower incidence of ischemic stroke compared to the aspirin group (OR = 0.49, 95% CrI 0.26-0.91). And the prolonged DAPT group had a higher total bleeding rate compared to aspirin group (OR = 2.4, 95% CrI 1.1-5.9). Conclusions: In terms of MACE, NACE, all-cause death, cardiac death, stent thrombosis, and major bleeding, there were no significant differences between using aspirin alone, clopidogrel alone, and ticagrelor alone; extending DAPT duration; and using aspirin combined with low-dose rivaroxaban for chronic maintenance antithrombotic regimens. However, choosing aspirin combined with low-dose rivaroxaban can reduce the incidence of ischemic stroke, and prolonged DAPT may have a higher rate of total bleeding. However, it is important to note that this study is based on indirect comparisons, and there is currently a lack of direct evidence comparing various maintenance antiplatelet therapy regimens. Further high-quality studies are needed to address this gap and provide more conclusive evidence on the comparative effectiveness of different maintenance antiplatelet strategies.

Left Bundle Branch Area Pacing versus Right Ventricular Pacing in Patients with Atrioventricular Block: An Observational Cohort Study.

Objective: We aim to conduct a comparison of the safety and effectiveness performance between left bundle branch area pacing (LBBAP) and right ventricular pacing (RVP) regimens for patients with atrioventricular block (AVB). Methods: This observational cohort study included patients who underwent pacemaker implantations with LBBAP or RVP for AVB indications from the 1st of January 2018 to the 18th of November 2021 at West China Hospital. The primary composite outcome included all-cause mortality, lead failure, or heart failure hospitalization (HFH). The secondary outcome included periprocedure complication, cardiac death, or recurrent unexplained syncope. A 1 : 1 propensity score-matched cohort was conducted for left ventricular (LV) function analysis. Results: A total of 903 patients met the inclusion criteria and completed clinical follow-up. After adjusting for the possible confounders, LBBAP was independently associated with a lower risk of the primary outcome (OR 0.48, 95% CI 0.28 to 0.83, p = 0.009), including a lower risk of all-cause mortality and HFH. No significant difference in the secondary outcome was detected between the groups except that LBBAP was independently associated with a lower risk of recurrent unexplained syncope. In the propensity-score matching cohort of echocardiographic analysis, the LV systolic dyssynchrony index was lower in LBBAP compared with that in RVP (5.68 ± 1.92 vs. 6.50 ± 2.28%, p = 0.012). Conclusions: Compared to conventional RVP, LBBAP is a feasible novel pacing model associated with a significant reduction in the primary composite outcome. Moreover, LBBAP significantly reduces the risk of recurrent unexplained syncope and improves LV systolic synchrony. This study is registered with ClinicalTrials.gov NCT05722379.

A portable test strip fabricated of luminescent lanthanide-functionalized metal-organic frameworks for rapid and visual detection of tetracycline antibiotics.

Tetracycline antibiotics (TCs) are commonly used antibiotics in the treatment of infections, but their overuse has a negative impact on human health and ecosystems. Thus, the development of a facile and on-site visualization method for TC detection is necessary. Here, we propose the potential of using lanthanide-functionalized metal-organic framework (MOF) composites (Ag+/Tb3+@UiO-66-(COOH)2, ATUC) as a probe for the rapid detection of tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC), and doxycycline (DOX) residues, in which UiO-66-(COOH)2 (UC) could be utilized to provide an interaction microenvironment, Tb3+ as recognition units and Ag+ as a fluorescence enhancer. Upon exposure to TCs, significant luminescence quenching of ATUC excited at 255 nm was observed due to the inner filter effect (IFE) and photo-induced electron transfer (PET), and the established strategy has a detection limit (LOD) of 11.0, 20.1, 9.1, and 22.5 nM for TC, CTC, OTC, and DOX, respectively. More importantly, given its portability and conspicuous luminescence color gradation variation, a portable test strip based on ATUC was manufactured and the results could be distinguished immediately by the naked eye and smartphone analysis, allowing for on-site rapid quantitative assay of TCs, not only in the laboratory but also in a point-of-care setting.

The effect of macromolecular crowding degree on the self-assembly of fatty acid and lipid hydrolysis.

Investigation on the physiochemical nature involved in the production of fatty acid catalyzed by the vesicles is of importance to understand the digestion of lipid. In this paper, the effects of crowding degree, which was constructed by polyethylene glycol (PEG), on the autocatalytic production of fatty acid with different chain lengths was studied. The results showed that the higher crowding degree led to the slower production rate of decanoic acid but the faster rate of oleic acid. The reason lies in that the presence of macromolecules resulted in the increased sizes of decanoic acid vesicles, but decreased sizes of oleic acid vesicles. Meanwhile, decanoic acid vesicles in more crowded medium exhibited viscous behavior, whereas oleic acid displayed elastic behavior. This research provides useful information for understanding the unusual autocatalyzed production of fatty acid in macromolecular crowding and may also draw an attention to the physiologically relevant lipid digestion.

The Management and Antithrombotic Strategies of Patients with Coronary Artery Disease and High Bleeding Risk.

PURPOSE OF REVIEW: In this review, we aim to summarize the current understanding of high bleeding risk (HBR) patients in coronary artery disease (CAD) and provide a comprehensive evaluation of the available antithrombotic strategies for both percutaneous coronary intervention (PCI) and coronary artery bypass grafting (CABG) procedures. RECENT FINDINGS: CAD is a major cause of mortality among cardiovascular diseases, resulting from insufficient blood flow in the coronary artery due to atherosclerosis. Antithrombotic therapy is a crucial component of drug therapy for CAD and multiple studies had been focusing on the optimal antithrombotic strategies of different CAD populations. However, there is no fully harmonized definition of the model of bleeding, and the optimal antithrombotic strategy for such patients at HBR is inconclusive. In this review, we summarize bleeding risk stratification models for CAD patients and discuss the de-escalation of antithrombotic strategies among HBR patients. Furthermore, we realize that for certain subgroups of CAD-HBR patients, more individualized and precise antithrombotic strategy development is needed. So, we highlight special populations, such as CAD patients combined with valvular diseases, with both high ischemia and bleeding risks, and those proceeding surgical treatment, which requires greater research attention. We note that de-escalating therapy for CAD-HBR patients is an emerging trend in managing this population, but the optimal antithrombotic strategies should be re-considered according to the patient's baseline characteristics.

Glutathione inhibited starch digestion: Structural and kinetic analysis of substrate and α-amylase.

Glutathione (GSH) is a natural antioxidant that helps fight free radicals. Whether it will affect the activity of amylase and starch digestion remains unknown. This research disclosed that GSH could interact with starch through hydrogen bonds, which accelerated the swelling of starch granules and promoted the formation of ordered double-helix crystalline, and therefore inhibited starch digestion. Moreover, pig pancreas α-amylase (PPA) which was incubated with GSH displayed a less stable conformation and decreased activity. However, in a crowded media constructed by sodium caseinate (NaCas), an antagonistic effect existed between GSH and NaCas. As the rate and extent of starch digestion have been linked with health aspects, this study suggests that GSH can be used in the formulation of diet foods. It also reminds us to consider the synergistic or antagonistic effects caused by the coexisted components in the complexed food matrix.

Effect of Ionic and Non-Ionic Surfactants on the Pasting Characteristics and Digestive Properties of Regular and Frozen Starch for Oral Delivery.

Starch is an ideal wall material for controlled release in oral delivery systems due to its non-allergic properties, availability, and cheap price. However, because of its poor mechanical behavior and high water permeability, it is necessary to modify the amphiphilic nature of starch. Surfactants are essential components to emulsify the lyophobic food ingredients. However, the interaction of starch with emulsifiers and how they affect the pasting behavior and digestion of starch are not well understood. In this paper, surfactants, such as non-ionic Tween (TW) and ionic sodium fatty acid (NaFA), with varying hydrophobic carbon chain lengths, were selected as model amphiphiles to investigate the structural, pasting, rheological properties and in vitro digestibility of regular and frozen starch samples. The results showed that, in most cases, the addition of TW reduced the viscosity of starch. However, saturated medium-chain NaFA increased the starch viscosity and rheological modulus greatly. Both surfactants inhibited starch digestion. This paper presents a comparative investigation on the effect of ionic and non-ionic surfactant on the structure and properties of corn starch, and therefore the information is useful for structural-based formulation with starch for developing colloidal delivery systems. It is also helpful for developing functional food with controllable digestion properties.

Soluble ST2 in coronary artery disease: Clinical biomarkers and treatment guidance.

The IL-33/ST2 L signaling pathway is involved in the pathophysiological processes of several diseases and mainly exerts anti-inflammatory and antifibrotic effects. Soluble suppression of tumorigenicity 2 (sST2), which serves as a competitive inhibitory molecule of this pathway, is a member of the interleukin (IL)-1 family, a decoy receptor for IL33, thought to play a role in cardiac remodeling and the inflammatory process. However, the association between sST2 and coronary artery disease (CAD), one of the most common causes of heart failure, is still being explored. We therefore reviewed the research on sST2 in the field of CAD, including reflecting the atherosclerosis burden, predicting no-reflow, predicting prognosis, responding to myocardial remodeling, and guiding management, hoping to provide cardiologists with new perspectives.

Short dual antiplatelet therapy in patients with high bleeding risk undergoing percutaneous coronary intervention: a systematic review and meta-analysis.

BACKGROUND: The efficacy and safety of an abbreviated duration of dual antiplatelet therapy (DAPT) in patients with high bleeding risk (HBR) undergoing percutaneous coronary intervention (PCI) (PCI-HBR patients) remain controversial. METHODS: The Cochrane Library, PubMed, EMBASE, and Ovid MEDLINE databases were searched. Studies that enrolled PCI-HBR patients as research subjects, compared different DAPT durations, and reported incidences of major adverse cardiac events (MACE) and net adverse clinical events (NACE) in PCI-HBR patients were obtained. The studies were stratified according to the DAPT duration (1, 3, and 6 months), and meta-analysis was subsequently performed. RESULTS: Nine studies (10 cohorts) were included in the meta-analysis. Compared with those who received DAPT for >1 month, PCI-HBR patients who received the 1-month DAPT regimen had comparable risks of NACE and MACE. Compared to those who received DAPT for >3 months, the risk of developing MACE in PCI-HBR patients who received the 3-month DAPT was not increased; however, the risk of ischemic stroke and stent thrombosis increased. Compared to those who received DAPT for >6 months, patients who received the 6-month DAPT had a reduction in the risk of major bleeding without an increase in NACE and MACE. CONCLUSIONS: Shortening the DAPT regimen to 1 or 6 months did not increase the risk of MACE, and the 6-month DAPT regimen reduced the risk of major bleeding. However, the 3-month DAPT regimen increased the risk of ischemic stroke. Thus, shortened DAPT reduced the risk of MACE and bleeding, with a small absolute increase in ischemic strokes.

Retrieving entrapped guidewire using rotablation technique: case series and literature review.

Background: The present article summarizes three cases of retrieving entrapped guidewire using rotablation technique. Case summary: All three cases pertained to guidewire being fractured and entrapped during percutaneous coronary interventions (PCIs) for right coronary artery (RCA) chronic total occlusion in a local hospital. In Case 1, two fractured filaments connecting to the entrapped guidewire could be clearly seen. One filament was in the right brachial artery and the other one was in the ascending aorta. In Case 2, the operator applied buddy wire technique during PCI but forgot to withdraw the second wire before stenting. As a result, the second SION wire was entrapped by the stent and was untangled and fractured when being retrieved. In Case 3, a SION guidewire was entrapped during PCI for RCA. The initial attempt to retrieve the guidewire ended in failure. Although the entrapped guidewire was still intact, it was difficult to advance the floppy rotawire into the RCA as it was still totally occluded. In those scenarios, complete retrieval of the entrapped guidewire using catheter-based technique was hardly possible. Rotablation was used to cut the entrapped guidewire, retrieve the broken end, and leave the remnants in coronary artery jailed by the stent. Clinical and angiographic follow-up observation demonstrated that the patients were free of target vessel failure. Discussion: Entrapment of guidewire is a rare complication of PCI. The management of entrapped guidewire is usually challenging. Rotablation technique is a useful and efficient strategy when conventional methods failed to retrieve the entrapped guidewire.

An artificial intelligence-based risk prediction model of myocardial infarction.

BACKGROUND: Myocardial infarction can lead to malignant arrhythmia, heart failure, and sudden death. Clinical studies have shown that early identification of and timely intervention for acute MI can significantly reduce mortality. The traditional MI risk assessment models are subjective, and the data that go into them are difficult to obtain. Generally, the assessment is only conducted among high-risk patient groups. OBJECTIVE: To construct an artificial intelligence-based risk prediction model of myocardial infarction (MI) for continuous and active monitoring of inpatients, especially those in noncardiovascular departments, and early warning of MI. METHODS: The imbalanced data contain 59 features, which were constructed into a specific dataset through proportional division, upsampling, downsampling, easy ensemble, and w-easy ensemble. Then, the dataset was traversed using supervised machine learning, with recursive feature elimination as the top-layer algorithm and random forest, gradient boosting decision tree (GBDT), logistic regression, and support vector machine as the bottom-layer algorithms, to select the best model out of many through a variety of evaluation indices. RESULTS: GBDT was the best bottom-layer algorithm, and downsampling was the best dataset construction method. In the validation set, the F1 score and accuracy of the 24-feature downsampling GBDT model were both 0.84. In the test set, the F1 score and accuracy of the 24-feature downsampling GBDT model were both 0.83, and the area under the curve was 0.91. CONCLUSION: Compared with traditional models, artificial intelligence-based machine learning models have better accuracy and real-time performance and can reduce the occurrence of in-hospital MI from a data-driven perspective, thereby increasing the cure rate of patients and improving their prognosis.

Improving Thermostability of Chimeric Enzymes Generated by Domain Shuffling Between Two Different Original Glucoamylases.

In order to improve enzymatic properties of glucoamylases, six recombinant genes GA1-GA6 were created by domain shuffling of glucoamylase genes GAA1 from Aspergillus niger Ld418AI and GATE from Talaromyces emersonii Ld418 TE using overlap extension PCR and were expressed in Saccharomyces cerevisiae W303-1B; only activities of GA1 and GA2 in the fermentation broth were higher than those of GAA1 but less than those of GATE. Further research results of GA1 and GA2 indicated that chimeric glucoamylases GA1 and GA2 revealed increased thermostability compared with GAA1 and GATE, although with a slight change in the activity and optimal temperature. However, GA1 had almost the same catalytic efficiency as GATE, whereas the catalytic efficiency of GA2 was slightly less than that of GATE, but still higher than that of GAA1. The structural analysis showed that the change of enzymatic properties could be caused by the increased and extended α-helix and ß-sheet, which change the secondary and tertiary structures of chimeric glucoamylases. These results demonstrated that domain shuffling was feasible to generate a chimeric enzyme with novel properties.

Diagnostic accuracy of CT-derived and angiogram-derived fractional flow reserve.

AIMS: Although accumulating evidence demonstrated that virtual fractional flow reserve (FFR) based on coronary computed tomography angiography (CCTA) (CT-FFR) or invasive coronary angiogram (ICA) (CA-FFR) are promising alternatives to wire based FFR, which method has better diagnostic accuracy was still unclear. In our study, we aim to directly compare the diagnostic performance of CT-FFR and CA-FFR. METHODS: During the period of September 2019 to December 2020, patients with at least one 30%-90% coronary artery stenosis were enrolled and received invasive FFR. Then, virtual FFR values were calculated based on both CCTA and ICA, and then compared with the invasive FFR value. RESULTS: Invasive FFR measurements were successfully performed in 114 vessels of 96 patients. Both CT-FFR and CA-FFR showed good correlation with wire-based FFR, with r values of 0.84 and 0.71 respectively. In paired t-test, the deviation of CT-FFR and CA-FFR was not significantly different (t = -1.9083, p = 0.05889). In Bland-Altman analysis, the coefficients of variation were 8.4% and 13.2% for CT-FFR and CA-FFR respectively. In ROC curve analysis, the per-vessel diagnostic accuracy of CT-FFR and CA-FFR was 94.7% and 92.1% respectively. The area under the curve of CT-FFR was slightly higher than that of CA-FFR (0.986 and 0.916 respectively, the difference between areas = 0.070, 95% CI 0.003-0.137, p = 0.0227). CONCLUSION: Both CT-FFR and CA-FFR had good diagnostic concordance with wire-based FFR. In ROC Curve analysis, CT-FFR demonstrated slightly higher diagnostic accuracy than CA-FFR. CLINICAL TRIAL REGISTRATION: URL: https://www.chictr.org.cn/showproj.aspx?proj=44719. Unique Identifier: ChiCTR1900026971.

l-Arginine inhibits the activity of α-amylase: Rapid kinetics, interaction and functional implications.

In this work, the rapid unfolding kinetics of pancreas α-amylase (PPA) induced by l-arginine and the interaction mechanism were investigated. The unfolding followed a first-level reaction kinetics equation, without intermediates. l-arginine interacted with PPA though diffusion-controlled process rather than complexion. The interaction between l-arginine and PPA resulted in a pronounced decrease in ß-sheet and a significant increase in random coil, and thereby the enzyme activity decreased. However, the unfolding of PPA could be compensated and the second structure change could be recovered to some extent by the macromolecular crowded medium of Pluronics. Further insight into the mechanism disclosed that the broken H-bond network of water may contribute to PPA unfolding. This work provides a new perspective on the interaction of l-arginine with digestive enzyme. The unfolding mechanism of enzymes by may help to understand the effects of other structurally similar drugs, which is of concern in food-drug interactions.

[An efficient marker-free genome editing method for Aspergillus niger].

Aspergillus niger is an important industrial strain which has been widely used for production of enzymes and organic acids. Genome modification of A. niger is required to further improve its potential for industrial production. CRISPR/Cas9 is a widely used genome editing technique for A. niger, but its application in industrial strains modification is hampered by the need for integration of a selection marker into the genome or low gene editing efficiency. Here we report a highly efficient marker-free genome editing method for A. niger based on CRISPR/Cas9 technique. Firstly, we constructed a co-expression plasmid of sgRNA and Cas9 with a replication initiation region fragment AMA1 (autonomously maintained in Aspergillus) by using 5S rRNA promoter which improved sgRNA expression. Meanwhile, a strain deficient in non-homologous end-joining (NHEJ) was developed by knocking out the kusA gene. Finally, we took advantage of the instability of plasmid containing AMA1 fragment to cure the co-expression plasmid containing sgRNA and Cas9 through passaging on non-selective plate. With this method, the efficiency of gene editing reached 100% when using maker-free donor DNA with a short homologous arm of 20 bp. This method may facilitate investigation of gene functions and construction of cell factories for A. niger.