Structural insights into the chloroplast protein import in land plants.

Chloroplast proteins are imported via the translocon at the outer chloroplast membrane (TOC)-translocon at the inner chloroplast membrane (TIC) supercomplex, driven by an ATPase motor. The Ycf2-FtsHi complex has been identified as the chloroplast import motor. However, its assembly and cooperation with the TIC complex during preprotein translocation remain unclear. Here, we present the structures of the Ycf2-FtsHi and TIC complexes from Arabidopsis and an ultracomplex formed between them from Pisum. The Ycf2-FtsHi structure reveals a heterohexameric AAA+ ATPase motor module with characteristic features. Four previously uncharacterized components of Ycf2-FtsHi were identified, which aid in complex assembly and anchoring of the motor module at a tilted angle relative to the membrane. When considering the structures of the TIC complex and the TIC-Ycf2-FtsHi ultracomplex together, it becomes evident that the tilted motor module of Ycf2-FtsHi enables its close contact with the TIC complex, thereby facilitating efficient preprotein translocation. Our study provides valuable structural insights into the chloroplast protein import process in land plants.

Continuous Strain Regulation of Palladium-Gold at the Atomic Level.

Revealing the effect of surface structure changes on the electrocatalytic performance is beneficial to the development of highly efficient catalysts. However, precise regulation of the catalyst surface at the atomic level remains challenging. Here, we present a continuous strain regulation of palladium (Pd) on gold (Au) via a mechanically controllable surface strain (MCSS) setup. It is found that the structural changes induced by the strain setup can accelerate electron transfer at the solid-liquid interface, thus achieving a significantly improved performance toward hydrogen evolution reaction (HER). In situ X-ray diffraction (XRD) experiments further confirm that the enhanced activity is attributed to the increased interplanar spacing resulting from the applied strain. Theoretical calculations reveal that the tensile strain modulates the electronic structure of the Pd active sites and facilitates the desorption of the hydrogen intermediates. This work provides an effective approach for revealing the relationships between the electrocatalyst surface structure and catalytic activity.

Superconductivity above 105 K in Nonclathrate Ternary Lanthanum Borohydride below Megabar Pressure.

Hydrides are promising candidates for achieving room-temperature superconductivity, but a formidable challenge remains in reducing the stabilization pressure below a megabar. In this study, we successfully synthesized a ternary lanthanum borohydride by introducing the nonmetallic element B into the La-H system, forming robust B-H covalent bonds that lower the pressure required to stabilize the superconducting phase. Electrical transport measurements confirm the presence of superconductivity with a critical temperature (Tc) of up to 106 K at 90 GPa, as evidenced by zero resistance and Tc shift under an external magnetic field. X-ray diffraction and transport measurements identify the superconducting compound as LaB2H8, a nonclathrate hydride, whose crystal structure remains stable at pressures as low as ∼ half megabar (59 GPa). Stabilizing superconductive stoichiometric LaB2H8 in a submegabar pressure regime marks a substantial advancement in the quest for high-Tc superconductivity in polynary hydrides, bringing us closer to the ambient pressure conditions.

Self-Organized Evolution of the Internal Transport Barrier in Ion-Temperature-Gradient Driven Gyrokinetic Turbulence.

Understanding the self-organization of the most promising internal transport barrier in fusion plasmas needs a long-time nonlinear gyrokinetic global simulation. The neighboring equilibrium update method is proposed, which solves the secularity problem in a perturbative simulation and speeds up the numerical computation by more than 10 times. It is found that the internal transport barrier emerges at the magnetic axis due to inward propagated turbulence avalanche, and its outward expansion is the catastrophe of self-organized structure induced by outward propagated avalanche.

Scaffold Hopping of Pristimerin Provides Derivatives Containing a Privileged Quinoxaline Substructure as Potent Autophagy Inducers in Breast Cancer Cells.

Pristimerin is a natural triterpenoid that has received much attention from medicinal chemists for its multiple biological activities. However, structural modifications of pristimerin, especially those aimed at discovering antitumor agents, are relatively limited. In this study, two series of pristimerin derivatives containing phenyloxazole and quinoxaline moieties, respectively, were designed via the scaffold hopping strategy. The target compounds were synthesized and analyzed for their cytotoxic activities in vitro using the MTT assay. The most potent cytotoxic compound (21o) significantly inhibited the proliferation of MCF-7 cells with an IC50 value of 2.0 µM, 1.5-fold more potent than pristimerin (IC50 = 3.0 µM). Compared with pristimerin, compound 21o displayed the greatest improvement in selectivity (25.7-fold) against the MCF-7 and MCF-10A cell lines. Transmission electron microscopy, monodansylcadaverine and DCFH-DA staining, Western blotting, and different inhibitor assays were performed to elucidate the mechanism of action of compound 21o. Compound 21o induced autophagy-mediated cell death in MCF-7 cells by activating the ROS/JNK signaling pathway. Therefore, incorporating a quinoxaline substructure into pristimerin could be advantageous for enhancing its cytotoxic activity. Compound 21o may serve as a lead compound for developing new therapies to treat breast cancer.

CDCA7 promotes TGF-ß-induced epithelial-mesenchymal transition via transcriptionally regulating Smad4/Smad7 in ESCC.

Cell division cycle associated 7 (CDCA7) is a copy number amplification gene that contributes to the metastasis and invasion of tumors, including esophageal squamous cell carcinoma (ESCC). This present study aimed at clarifying whether high expression of CDCA7 promotes the metastasis and invasion of ESCC cell lines and exploring the underlying mechanisms implicated in epithelial-mesenchymal transition (EMT) of ESCC. The role of CDCA7 in the regulation of ESCC metastasis and invasion was evaluated using ESCC cell lines. Expression of EMT-related markers including E-cadherin, N-cadherin, Vimentin, Snail, and Slug, transforming growth factor ß (TGF-ß) signaling pathway including Smad2/3, p-Smad2/3, Smad4, and Smad7 were detected in CDCA7 knockdown and overexpressed cell lines. Dual-luciferase reporter assay and rescue assay were used to explore the underlying mechanisms that CDCA7 contributed to the metastasis and invasion of ESCC. High CDCA7 expression significantly promoted the metastasis and invasion of ESCC cell lines both in vivo and in vitro. Additionally, the expression of CDCA7 positively correlated with the expression of N-cadherin, Vimentin, Snail, Slug, TGF-ß signaling pathway and negatively correlated with the expression of E-cadherin. Furthermore, CDCA7 transcriptionally regulated the expression of Smad4 and Smad7. Knockdown of CDCA7 inhibited the TGF-ß signaling pathway and therefore inhibited EMT. Our data indicated that CDCA7 was heavily involved in EMT by regulating the expression of Smad4 and Smad7 in TGF-ß signaling pathway. CDCA7 might be a new therapeutic target in the suppression of metastasis and invasion of ESCC.

Inner filter effect-based near-infrared fluorescent probe for detection of metronidazole on a smartphone-integrated analytical platform.

Antibiotic residues pose a serious threat to ecosystems and food safety. Developing convenient, visual, and on-site detection methods is therefore in high demand and has a practical purpose. In this work, a near-infrared (NIR) fluorescent probe with an analysis platform based on a smartphone has been constructed for quantitative and on-site detection of metronidazole (MNZ). CdTe quantum dots with NIR emission at 710 nm (QD710) were prepared by using a simple hydrothermal method and showed good properties. A spectral overlap between absorption of MNZ and excitation of QD710 resulted in an effective inner filter effect (IFE) between QD710 and MNZ. Because of the IFE, the fluorescence of QD710 decreased gradually with increasing concentrations of MNZ. Based on the fluorescence response, quantitative detection and visualization of MNZ was achieved. NIR fluorescence analysis and the special IFE between probe and target can improve sensitivity and selectivity for MNZ. Additionally, these were also utilized for quantitative detection of MNZ in real food samples and the results were reliable and satisfactory. Meanwhile, a portable visual analysis platform in a smartphone was constructed for on-site analysis of MNZ, which can be used as an alternative method for detection of MNZ residues in situations with limited instrumental conditions. Therefore, this work provides a convenient, visual, and real-time analysis method for detection of MNZ and the analysis platform shows great potential for commercialization.

Family history, waist circumference and risk of ischemic stroke: A prospective cohort study among Chinese adults.

BACKGROUND AND AIMS: The associations between genetic factors and waist circumference (WC) with stroke risk have been evaluated in Western studies. However, evidence of this association has rarely been reported in the Chinese population. This study aimed to evaluate the association between WC and family history of stroke (FHS) with ischemic stroke (IS) risk among Chinese adults and to further explore the potential interaction of these associations. METHODS AND RESULTS: The China Kadoorie Biobank (CKB) study recruited 35,508 participants aged 30-79 years from the Qingdao urban area during 2004-2008. A total of 33,355 participants were included in study. Cox regression analysis was used to estimate the multivariable-adjusted hazard ratios (HR) and 95% confidence intervals (CI) for the independent and interactional associations between FHS and WC and IS risk. Participants with FHS had a 29% (HR = 1.29, 95% CI: 1.12-1.50) higher IS risk than those without FHS. Participants with excessive WC (85 cm for males and 80 cm for females) had a 78% (HR = 1.78, 95% CI: 1.51-2.10) higher IS risk than those with normal WC. The combined effect of FHS and excessive WC on IS was statistically significant (HR = 2.29, 95% CI: 1.84-2.86). The present study further found statistically significant multiplicative interactions of FHS and WC with IS risk (Pinteraction < 0.001). CONCLUSION: The present study indicated that FHS and WC were significantly associated with an increased risk of IS. The association between FHS and IS was associated with excessive WC.

AP-HTPB propellant combustion under strain conditions with laser absorption spectroscopy.

Understanding the combustion behaviors of solid propellant with different levels of strains is of practical interest. In this work, an experimental study of the effects of static and dynamic strains on the burning rate, temperature, CO, and C O 2 formation of aluminized ammonium perchlorate (AP)-hydroxyl terminated poly-butadiene (HTPB) propellant combustion was presented at initial pressures of 0.1 MPa, 0.2 MPa, and 0.5 MPa. The strains were being applied onto solid propellant by exerting static and cyclic loadings. The propellant burning rate was acquired by a 4 kHz high-speed photography system, and the combustion temperature, CO, and C O 2 column densities were measured at 10 kHz through laser absorption spectroscopy (LAS). At atmospheric pressure, it was demonstrated that the propellant burning rate increased with tensile stress and decreased with compressive stress. The measured flame temperature showed a similar correlation with strains as compared to the propellant burning rate. At elevated pressures, the increase of the propellant burning rate due to tensile stress was more evident, while the difference in combustion temperatures was less significant. For the cyclic strain condition, the variations of the measured C O 2 and CO column densities were consistent with the static strain condition.

Carbon dot and silver nanoparticle-based fluorescent probe for the determination of sulfite and bisulfite via inner-filter effect and competitive redox reactions.

A new sensitive fluorescent probe (CDs-AgNP/H2O2) for detecting sulfite and bisulfite (SO32- and HSO3-) based on the inner-filter effect (IFE) between silver nanoparticles (AgNPs) and carbon dots (CDs) was developed. Because of the spectral overlap between the absorption of AgNPs and the excitation of CDs, the fluorescence of CDs can be quenched by AgNPs owing to the IFE. H2O2 weakens the IFE and restores the fluorescence due to the oxidation of AgNPs by H2O2. However, the existence of SO32-/HSO3- can quench the fluorescence again as a result of redox reaction between SO32-/HSO3- and H2O2. The results showed a broad linear range of 20-200 µM with a low limit of detection (3.02 µM) toward SO32-/HSO3-. The combination of IFE and redox reaction led to improvement of the sensitivity and selectivity. The probe was implemented to measure SO32-/HSO3- in various agricultural products and foods with acceptable results (80.6 to 118.9% recovery).

A Dielectric Polymer/Metal Oxide Nanowire Composite for Self-Adaptive Charge Release.

Electrostatic discharge is a fatal threat to a variety of electronic products. Here we report on a polymer nanocomposite composed of a dielectric polymer embedded with aligned core-shell structured nanowires for highly efficient distributed electrostatic discharge protection. The dielectric nanocomposite is capable of self-adaptive charge release, stemming from the nonlinear interface built in the Bi/Co oxide coated ZnO nanowires that leads to a "hand-in-hand" double-Schottky barrier. The ultralow filler concentration (e.g., 0.5 vol %) endows the nanocomposite with low permittivity (close to the pristine polymer matrix) and high optical transmittance (75%) in the visible light wavelength range, which is desirable in packaging materials and display coatings for portable electronics. The effectiveness for electrostatic discharge protection was validated with an organic light-emitting diode screen protected by the nanocomposite that survived multiple rounds of electrostatic discharge.

Engineering diverse fatty acid compositions of phospholipids in Escherichia coli.

Bacterial fatty acids (FAs) are an essential component of the cellular membrane and are an important source of renewable chemicals as they can be converted to fatty alcohols, esters, ketones, and alkanes, and used as biofuels, detergents, lubricants, and commodity chemicals. Most prior FA bioconversions have been performed on the carboxylic acid group. Modification of the FA hydrocarbon chain could substantially expand the structural and functional diversity of FA-derived products. Additionally, the effects of such modified FAs on the growth and metabolic state of their producing cells are not well understood. Here we engineer novel Escherichia coli phospholipid biosynthetic pathways, creating strains with distinct FA profiles enriched in ω7-unsaturated FAs (ω7-UFAs, 75%), Δ5-unsaturated FAs (Δ5-UFAs, 60%), cyclopropane FAs (CFAs, 55%), internally-branched FAs (IBFAs, 40%), and Δ5,ω7-double unsaturated FAs (DUFAs, 46%). Although bearing drastically different FA profiles in phospholipids, UFA, CFA, and IBFA enriched strains display wild-type-like phenotypic profiling and growth. Transcriptomic analysis reveals DUFA production drives increased differential expression and the induction of the fur iron starvation transcriptional cascade, but higher TCA cycle activation compared to the UFA producing strain. This likely reflects a slight cost imparted for DUFA production, which resulted in lower maximum growth in some, but not all, environmental conditions. The IBFA-enriched strain was further engineered to produce free IBFAs, releasing 96 mg/L free IBFAs from 154 mg/L of the total cellular IBFA pool. This work has resulted in significantly altered FA profiles of membrane lipids in E. coli, greatly increasing our understanding of the effects of FA structure diversity on the transcriptome, growth, and ability to react to stress.

Insight into the Experimental Error in the Mapping of Electrical Properties with Electrostatic Force Microscopy.

Electrostatic force microscopy (EFM) is an emergent, powerful technique for nanoscale detection of electrical properties such as permittivity and charge distribution. However, the surface irregularity of samples has been unfortunately overlooked in most EFM studies. Herein, we use a polymer nanocomposite dielectric (PND) as the showcase and demonstrate that the morphological discontinuity at the matrix/particle interface can lead to major discrepancies or even incorrect results in the EFM study. First, the influence of the morphology, permittivity, and charge density of the interface is quantitively analyzed with a numerical method, proving that linking EFM results directly to sample properties is impracticable in the research based on classical interface configuration. Then, two methods are proposed to address the issue. The first method is numerical inversion, which takes heterogeneous materials and irregular surfaces into consideration. In this method, the influence of several experimental uncertainties, such as the radius of the nanoparticle and the permittivity of the matrix, is estimated. It is shown that the uncertainties related to geometry have a great impact on inversion and should be determined preferentially. In the second method, two standard configurations of the interface are recommended and compared for the interface study to bypass the morphological issue. This work provides quantitative results regarding the long-overlooked error in the EFM detection of the microregion with heterogeneous composition and surface irregularities and offers methods to tackle this issue.

Thiazolidinedione-Based Structure Modification of Celastrol Provides Thiazolidinedione-Conjugated Derivatives as Potent Agents against Non-Small-Cell Lung Cancer Cells through a Mitochondria-Mediated Apoptotic Pathway.

In order to improve the potential of celastrol against non-small-cell lung cancer cells, the privileged structure, thiazolidinedione, was introduced into its C-20 carboxylic group with acetylpiperazine as a linker, and the thiazolidinedione-conjugated compounds 10a-10t were prepared. The target compounds were evaluated for their cytotoxic activities against the A549 cell line, and the results showed that most of the compounds 10a-10t displayed improved potency over celastrol, and compound 10b exhibited significant activity against the A549 cell line, with an IC50 value of 0.08 µM, which was 13.8-fold more potent than celastrol (IC50 = 1.10 µM). The mechanistic studies suggested that 10b could induce A549 cell apoptosis, as evidenced by Hoechst 33342 staining and annexin V-FITC/propidium iodide dual staining assays. Western blot analysis suggested that compound 10b could upregulate Bax expression, downregulate Bcl-2 expression, and activate the mitochondria-mediated apoptotic pathway. Furthermore, compound 10b could effectively inhibit tumor growth when tested in an A549 cell xenograft mouse model. Collectively, compound 10b is worthy of further investigation to support the discovery of effective agents against non-small-cell lung cancer.

Modeling and Verification of an Acquisition Strategy for Wheel Loader's Working Trajectories and Resistance.

To overcome the difficulty of collecting the working resistance and working trajectory of a wheel loader, this paper constructs a statics model of the bucket working resistance and a kinematics model of the working trajectory in the shoveling process and analyzes the key parameters of measuring the working resistance and working trajectory. Based on this, a working resistance and working trajectory acquisition strategy is proposed. To verify the effectiveness of the acquisition strategy, the in-service operation data of fine sand and loose soil shoveled by the wheel loader are collected and analyzed. Then, the test-fitted working resistance and working trajectory are obtained, and the working trajectory is input into the RecurDyn-EDEM co-simulation model to obtain the simulation-fitted working resistance. Considering the complex working conditions of the wheel loader, it is difficult to obtain accurate working resistance, and the actual working resistance is also a relative value. Therefore, a strong correlation between the two curves indicates that the acquisition strategy of the wheel loader's working trajectory and working resistance proposed in this paper is feasible.

Multi-Object Detection Method in Construction Machinery Swarm Operations Based on the Improved YOLOv4 Model.

To handle the problem of low detection accuracy and missed detection caused by dense detection objects, overlapping, and occlusions in the scenario of complex construction machinery swarm operations, this paper proposes a multi-object detection method based on the improved YOLOv4 model. Firstly, the K-means algorithm is used to initialize the anchor boxes to improve the learning efficiency of the depth features of construction machinery objects. Then, the pooling operation is replaced with dilated convolution to solve the problem that the pooling layer reduces the resolution of feature maps and causes a high missed detection rate. Finally, focus loss is introduced to optimize the loss function of YOLOv4 to improve the imbalance of positive and negative samples during the model training process. To verify the effectiveness of the above optimizations, the proposed method is verified on the Pytorch platform with a self-build dataset. The experimental results show that the mean average precision(mAP) of the improved YOLOv4 model for multi-object detection of construction machinery can reach 97.03%, which is 2.16% higher than that of the original YOLOv4 detection network. Meanwhile, the detection speed is 31.11 fps, and it is reduced by only 0.59 fps, still meeting the real-time requirements. The research lays a foundation for environment perception of construction machinery swarm operations and promotes the unmanned and intelligent development of construction machinery swarm operations.

Construction of Highly Active Zn3In2S6 (110)/g-C3N4 System by Low Temperature Solvothermal for Efficient Degradation of Tetracycline under Visible Light.

Herein, Zn3In2S6 photocatalyst with (110) exposed facet was prepared by low temperature solvothermal method. On this basis, a highly efficient binary Zn3In2S6/g-C3N4 was obtained by low temperature solvothermal method and applied to the degradation of tetracycline (TC). The samples of the preparation were characterized by X-ray diffraction, scanning electron microscope, transmission electron microscope, UV-vis diffuse reflection spectroscopy, and photoluminescence spectroscopy. Furthermore, the degradation performance of photocatalysts on TC was investigated under different experimental conditions. Finally, the mechanism of Zn3In2S6/g-C3N4 composite material degrading TC is discussed. The results show that Zn3In2S6 and Zn3In2S6/g-C3N4 photocatalysts with excellent performance could be successfully prepared at lower temperature. The Zn3In2S6/g-C3N4 heterojunction photocatalyst could significantly improve the photocatalytic activity compared with g-C3N4. After 150 min of illumination, the efficiency of 80%Zn3In2S6/g-C3N4 to degrade TC was 1.35 times that of g-C3N4. The improvement of photocatalytic activity was due to the formation of Zn3In2S6/g-C3N4 heterojunction, which promoted the transfer of photogenerated electron-holes. The cycle experiment test confirmed that Zn3In2S6/g-C3N4 composite material had excellent stability. The free radical capture experiment showed that ·O2- was the primary active material. This study provides a new strategy for the preparation of photocatalysts with excellent performance at low temperature.

Artificially constructing mixed bacteria system for bioaugmentation of nitrogen removal from saline wastewater at low temperature.

To alleviate the inhibition effects of multi-stresses, a multi-bacterial bioaugmentation based on stimulating cell-to-cell interactions was applied to improve the stress potential of salt-tolerant aerobic granular sludge (AGS). Results showed that the consortium formed by a combination of salt-tolerant ammonia-nitrogen utilizing bacteria, salt-tolerant nitrite-nitrogen utilizing bacteria and salt-tolerant nitrate-nitrogen utilizing bacteria with a whole biomass ratio of 1:2:1 achieved maximum nitrogen consumption rate (µNH4+-N, µNO2--N and µNO3--N of 1.03, 0.57 and 11.62 mgN/L·h, respectively) at 35 gNaCl/L salinity and 15 °C. The flocculent consortium was aggregated by Aspergillus tubingensis mycelium pellet, which was made into a compound bacterial agent (CBA), and the comprehensive nitrogen consumption capability of CBA was further improved to 2.47-4.36-fold of single functional bacteria. 5% CBA (m/m) was introduced into the seafood processing wastewater in batches, in winter (12-16 °C), the removal efficiencies of NH4+-N and total nitrogen increased from 66.89% to 52.77% of native AGS system to 79.02% and 69.97% of nascent bioaugmentation system, respectively. The analysis of key enzyme activities demonstrated that the ammonia monooxygenase and nitrate reductase activities of the bioaugmentation system were increased to 2.73-folds and 1.94-folds those of the native system. Moreover, due to an increase of 6.18 mg/gVSS and 0.11 in the secreted exopolysaccharide and tightly-bound/total extracellular polymeric substances, respectively, bioaugmentation boosted the cell bioflocculation ability by 13.53%, which enhanced the robustness. This work provided a detailed and feasible technical proposal for enhancing the biological treatment performance of saline wastewater in cold regions.

Adaptive regulation of activated sludge's core functional flora based on granular internal spatial microenvironment.

A great deal of efforts has been put into studying the influence of the external macroenvironment for activated sludge to survive on microbial community succession, while granular internal spatial microenvironment should be given equal attention, because it is more directly involved in the information exchange and material transfer among microorganisms. This study systematically investigated the effects of granular microenvironment on spatial colonization and composition of sludge's core functional flora, and the corresponding difference of biological treatment performance. High content of extracellular-proteins (67.53 mg/gVSS) or extracellular-polysaccharide (65.02 mg/gVSS) stimulated the microbial flocculation and aggregation of 0.5-1.5 mm granules (GS) or 1.5-3.0 mm granules (GM), respectively, which was resulted from excellent cell hydrophobicity (59.26%) or viscosity (3.47 mPa s), therefore, constituted relatively dense porous frame. More hollow space existed in 3.0-5.0 mm granules (GL), which formed loose skeleton with 0.213 mL/g of total pore volume and 17.21 nm of average pore size. Combining scanning electron microscope images and fluorescent in-situ hybridization based microbiological analysis, aerobic nitrifiers were observed to wrap or surround anaerobic bacteria, or facultative/anaerobic bacteria were self-encapsulated, which created granule's unique microenvironment with alternating aerobic and anaerobic zones. GS has the most rich organic matter degrading bacteria and anaerobic heterotrophic denitrifiers, while GM and GL presented the greatest relative abundance of facultative and aerobic denitrifiers, respectively. The activity of dehydrogenase and nitrogen invertase of GM showed be 1.32-3.09 times higher than those of GS and GL, contributing to its higher carbon and nitrogen removal. These findings highlight the importance of granular microenvironment to adaptive regulation of activated sludge's core functional flora and corresponding pollutant removal performance.

Role of initial bacterial community in the aerobic sludge granulation and performance.

The structure of bacterial community was greatly varied from different seed sludge sources, which affected the sludge characteristics. To explore the role of different functional bacteria in AGS granulation and pollutant degradation, three different resources of seed sludge obtained from pharmaceutical wastewater (R1), livestock (R2), and municipal sludge (R3) were employed in this study. Results showed that the initial bacterial community had important significance for AGS formation and pollutants removal. Seed sludge taken from R3 granulated faster than those from R1 and R2. A large number of mature granules were formed after 20 days of operation in R3. In addition, the final mixed liquor suspended solids (MLSS) reached 6853 mg L-1, with 48 mL g-1 sludge volume index (SVI) in R3, indicating that it had better settling performance and granulation. In the stable stage of R3, the removal rates of COD, NH4+-N, and TN reached 99.2%, 98.5%, and 97.6%, respectively. The α-diversity analysis showed that the bacterial community of seed sludge greatly determined the microbial composition of AGS. Firmicutes, Gracilibacteria, and Spirochaetes were abundant in R3, which maintained the structures and functions of aerobic granules. This study might provide approaches and insights for AGS culture from different sludge sources.