Polyphenols alleviate metabolic disorders: the role of ubiquitin-proteasome system.

Metabolic disorders include obesity, nonalcoholic fatty liver disease, insulin resistance and type 2 diabetes. It has become a major health issue around the world. Ubiquitin-proteasome system (UPS) is essential for nearly all cellular processes, functions as a primary pathway for intracellular protein degradation. Recent researches indicated that dysfunctions in the UPS may result in the accumulation of toxic proteins, lipotoxicity, oxidative stress, inflammation, and insulin resistance, all of which contribute to the development and progression of metabolic disorders. An increasing body of evidence indicates that specific dietary polyphenols ameliorate metabolic disorders by preventing lipid synthesis and transport, excessive inflammation, hyperglycemia and insulin resistance, and oxidative stress, through regulation of the UPS. This review summarized the latest research progress of natural polyphenols improving metabolic disorders by regulating lipid accumulation, inflammation, oxidative stress, and insulin resistance through the UPS. In addition, the possible mechanisms of UPS-mediated prevention of metabolic disorders are comprehensively proposed. We aim to provide new angle to the development and utilization of polyphenols in improving metabolic disorders.

Microbial community assemblage altered by coprecipitation of artificial humic substances and ferrihydrite: Implications for carbon fixation pathway transformation.

The suppression of soil carbon mineralization has been demonstrated to be effectively facilitated by carbon­iron interactions, yet the specific mechanisms by which artificial humic substances (A-HS) coupled with ferrihydrite influence this process remain insufficiently explored. This study is to investigate how the A-HS, specifically artificial fulvic acid (A-FA) and artificial humic acid (A-HA), coupled with ferrihydrite, affect carbon mineralization under anaerobic system that simulates paddy flooding conditions. The object is to investigate trends in carbon emissions and to delineate microbial community structure and functional pathways. The findings indicate that A-HA and A-FA substantially reduce CO2 and CH4 emissions, with A-FA having a particularly pronounced effect on carbon fixation, halving CO2 concentrations. The low concentration of Fe(II) observed suggest that A-FA and A-HA impede the dissimilatory iron reduction (DIR) process. Detailed 16S rDNA sequencing and gene prediction analyses reveal changes in microbial community structures and functions, highlighting Methanobacterium as the dominant hydrogenotrophic methanogens. The reductive citric acid cycle, predominantly utilized by Clostridium carboxidivorans, was identified as the principal carbon fixation pathway. This work provides a novel insight into the microbial mechanisms of carbon sequestration and highlights the potential of A-HS in improving soil fertility and contributing to climate change mitigation through enhancing soil carbon storage.

Synergistic enhancement of cadmium immobilization and soil fertility through biochar and artificial humic acid-assisted microbial-induced calcium carbonate precipitation.

Microbially induced carbonate precipitation (MICP) is emerging as a favorable alternative to traditional soil remediation techniques for heavy metals, primarily due to its environmental friendliness. However, a significant challenge in using MICP for farmland is not only to immobilize heavy metals but also to concurrently enhance soil fertility. This study explores the innovative combination of artificial humic acid (A-HA), biochar (BC), and Sporosarcina pasteurii (S. pasteurii) to mitigate the bioavailability of cadmium (Cd) in contaminated agricultural soils through MICP. X-ray diffraction (XRD) and scanning electron microscope (SEM) analyses revealed that the integration of BC and A-HA significantly enhances Cd immobilization efficiency by co-precipitating with CaCO3. Moreover, this treatment also improved soil fertility and ecological functions, as evidenced by increases in total nitrogen (TN, 9.0-78.2 %), alkaline hydrolysis nitrogen (AN, 259.7-635.5 %), soil organic matter (SOM, 18.1-27.9 %), total organic carbon (TOC, 43.8-48.8 %), dissolved organic carbon (DOC, 36.0-88.4 %) and available potassium (AK, 176.2-193.3 %). Additionally, the relative abundance of dominant phyla such as Proteobacteria and Firmicutes significantly increased with the introduction of BC and A-HA in MICP. Consequently, the integration of BC and A-HA with MICP offers a promising solution for remediating Cd-contaminated agricultural soil and synergistically enhancing soil fertility.

Quaternized antimicrobial peptide mimics based on harmane as potent anti-MRSA agents by multi-target mechanism covering cell wall, cell membrane and intracellular targets.

Infectious disease caused by methicillin-resistant Staphylococcus aureus (MRSA) seriously threatens public health. The design of antimicrobial peptide mimics (AMPMs) based on natural products (NPs) is a new strategy to kill MRSA and slow the development of drug resistance recently. Here, we reported the design and synthesis of novel AMPMs based on harmane skeleton. Notably, compound 9b exhibited comparable or even better anti-MRSA activity in vitro and in vivo with minimum inhibitory concentration (MIC) of 0.5-2 µg/mL than the positive drug vancomycin. The highly active compound 9b not only showed low cytotoxicity, no obvious hemolysis and good plasma stability, but also presented low tendency of developing resistance. Anti-MRSA mechanism revealed that compound 9b could destroy cell wall structure by interacting with lipoteichoic acid and peptidoglycan, cause membrane damage by depolarization, increased permeability and destructed integrity, reduce cell metabolic activity by binding to lactate dehydrogenase (LDH), interfere cellular redox homeostasis, and bind to DNA. Overall, compound 9b killed the MRSA by multi-target mechanism, which provide a promising light for combating the growing MRSA resistance.

Lattice matching and halogen regulation for synergistically induced large Li and Na storage by halogenated MXene V3C2Cl2.

Suffering from the formation of metal-ion dendrites and low storage capacity, MXene materials exhibit unsatisfactory performance in Li and Na storage. In this study, we demonstrate that the MXene V3C2Cl2 structure can induce uniform Li and Na deposition. This is achieved through coherent heterogeneous interface reconstruction and regulated ion tiling by halogen surface termination. The high lattice matching (91% and 99%) between MXenes and Li/Na, along with positive Cl terminal regulation, guides Li/Na ions to nucleate uniformly on the V3C2Cl2 MXene matrix and grow in a planar manner. Cl termination proves effective in regulating Li/Na ions due to its moderate adsorption and diffusion coefficients. Furthermore, upon adsorption onto the Cl-terminated V3C2Cl2 monolayer, Li4 and Na4 clusters undergo dissociation, favoring uniform adsorption over cluster adsorption. V3C2Cl2 MXenes exhibit impressive Li/Na storage capacities of 434.07 mA h g-1 for Li and 217.03 mA h g-1 for Na, surpassing the Li storage capacity of Ti3C2Cl2 by three-fold and the Na storage capacity of V2C by 1.4 times. This study highlights the regulatory role of Cl surface terminals in dendrite formation and Li/Na ion deposition, with potential applications to other metal-ion storage electrodes.

Symmetry-breaking-induced ferroelectric HfSnX3 monolayers and their tunable Janus structures: promising candidates for photocatalysts and nanoelectronics.

Designing novel two-dimensional (2D) ferroelectric materials by symmetry breaking and studying their mechanisms play important roles in the discovery of new ferroelectric photocatalysts and nanoelectronics. In this study, we have systematically investigated a series of novel ferroelectric 2D HfSnX3 (X = S, Se and Te) monolayers through first-principles calculations. We found that each HfSnX3 monolayer contains a stable ferroelectric phase (FP) and a paraelectric phase (PP). The large polarization (up to 1.64 µC cm-2) in the FP can significantly bend the oxidation reduction potential of water, making HfSnX3 monolayers become excellent ferroelectric photocatalysts. Specifically, by designing a Janus structure to break the symmetry of the PP, we have excitingly obtained a stable Hf2GeSnSe6 (referred to as HGSS) monolayer with triple polarized states. HGSS not only possesses great visible light absorption properties (about 3 × 105 cm-1) as photocatalysts but also successfully solves the dead layer problem previously reported in practical applications. In addition, by constructing a heterostructure with graphene, HGSS has great application in the design of controllable ultrathin p-n junctions. Overall, our study not only predicts a series of potential ferroelectric photocatalytic materials, but also provides valuable insights for designing tunable polarized materials and nanoelectronics.

Artificial humic acid regulates the impact of fungal community on soil macroaggregates formation.

Artificial humic acid (A-HA), which is synthesized from agricultural wastes and has high similarity to a natural humic substance (HS) extracted from soil, has been proven by our group to have potential for biological carbon sequestration in black soils. However, the mechanism involves in the application of A-HA on soil aggregation processes resulting from microbial activity stimulation and modifications to microbial communities remains unclear. This study investigates the correlation between the formation and stability of soil aggregates and fungal communities with various amounts of A-HA added to the rhizosphere and non-rhizosphere soil. A-HA can increase the total organic carbon (TOC) and dissolved organic carbon (DOC) concentrations in soil, promoting macroaggregate formation and increasing the mean weight diameter (MWD). In addition, soil aggregate binding agents such as polysaccharides, protein, extracellular polymeric substances (EPS), and glomalin-related soil protein (GRSP) are significantly increased by the addition of A-HA. A-HA can drive microaggregate to assemble into macroaggregate by increasing the abundance of beneficial fungi (e.g., Trichoderma and Mortierella). The co-occurrence network supports that A-HA shifted the key species and increased interactions of fungal taxa. This study will lay a solid foundation for sustainable agricultural development of A-HA application for soil fertility restoration in the future.

The Molecular Basis of the Anti-Inflammatory Property of Astragaloside IV for the Treatment of Diabetes and Its Complications.

Astragali Radix is a significant traditional Chinese medication, and has a long history of clinical application in the treatment of diabetes mellitus (DM) and its complications. AS-IV is an active saponin isolated from it. Modern pharmacological study shows that AS-IV has anti-inflammatory, anti-oxidant and immunomodulatory activities. The popular inflammatory etiology of diabetes suggests that DM is a natural immune and low-grade inflammatory disease. Pharmacological intervention of the inflammatory response may provide promising and alternative approaches for the prevention and treatment of DM and its complications. Therefore, this article focuses on the potential of AS-IV in the treatment of DM from the perspective of an anti-inflammatory molecular basis. AS-IV plays a role by regulating a variety of anti-inflammatory pathways in multiple organs, tissues and target cells throughout the body. The blockade of the NF-κB inflammatory signaling pathway may be the central link of AS-IV's anti-inflammatory effect, resulting in a reduction in the tissue structure and function damage stimulated by inflammatory factors. In addition, AS-IV can delay the onset of DM and its complications by inhibiting inflammation-related oxidative stress, fibrosis and apoptosis signals. In conclusion, AS-IV has therapeutic prospects from the perspective of reducing the inflammation of DM and its complications. An in-depth study on the anti-inflammatory mechanism of AS-IV is of great significance for the effective use of Chinese herbal medicine and the promotion of its status and influence on the world.

Effects of different concentrations of butyrate on microbial community construction and metabolic pathways in anaerobic digestion.

Investigating the effect of butyric acid concentration on anaerobic digestion systems in complex systems is important for the efficient degradation of butyric acid and improving the efficiency of anaerobic digestion. In this study, different loadings of butyric acid with 2.8, 3.2, and 3.6 g/(L·d) were added to the anaerobic reactor. At a high organic loading rate of 3.6 g/(L·d), methane was efficiently produced with VBP (Volumetric Biogas Production) of 1.50 L/(L·d) and biogas content between 65% and 75%. VFAs concentration remained below 2000 mg/L. Metagenome sequencing revealed changes in the functional flora within different stages. Methanosarcina, Syntrophomonas, and Lentimicrobium were the main and functional microorganisms. That the relative abundance of methanogens exceeded 35% and methanogenic metabolic pathways were increased indicated the methanogenic capacity of the system significantly improved. The presence of a large number of hydrolytic acid-producing bacteria also indicated the importance of the hydrolytic acid-producing stage in the system.

Combinatorial Application of Papain and CD66B for Isolating Glioma- Associated Neutrophils.

BACKGROUND: Stromal cells in the tumor microenvironment play crucial roles in glioma development. Current methods for isolating tumor-associated stromal cells (such as neutrophils) are inefficient due to the conflict between tissue dissociation and cell surface protein protection, which hampers the research on patient-derived stromal cells. Our study aims to establish a novel method for isolating glioma-associated neutrophils (GANs). METHODS: To observe neutrophil-like polymorphonuclear cells, we performed Hematoxylin-Eosin staining on glioma tissues. For isolating single cells from glioma tissues, we evaluated the efficiency of tissue dissociation with FastPrep Grinder-mediated homogenization or proteases (trypsin or papain) digestion. To definite specific markers of GANs, fluorescence-activated cell sorting (FACS) and immunofluorescence staining were performed. FACS and Ficoll were performed for the separation of neutrophils from glioma tissue-derived single-cell or whole blood pool. To identify the isolated neutrophils, FACS and RT-PCR were carried out. RESULTS: Neutrophil-like cells were abundant in high-grade glioma tissues. Among the three tissue dissociation methods, papain digestion produced a 5.1-fold and 1.7-fold more living cells from glioma mass than physical trituration and trypsin digestion, respectively, and it preserved over 97% of neutrophil surface protein markers. CD66B could be adopted as a unique neutrophil surface protein marker for FACS sorting in glioma. Glioma-derived CD66B+ cells specifically expressed neutrophil marker genes. CONCLUSION: A combination of papain-mediated tissue dissociation and CD66B-mediated FACS sorting is an effective novel method for the isolation of GANs from glioma tissues.

Efficacy and safety of Tonifying Qi and activating blood Chinese herbal prescriptions for myocardial infarction: Study protocol for a multi-centered RCT.

INTRODUCTION: Acute myocardial infarction (AMI) is a common cause of death worldwide and heart failure (HF) is the main complication. Although the increase in percutaneous coronary intervention and drug treatment can reduce in-hospital mortality after AMI, the incidence of HF after AMI and the resulting risk of death are still rising, which causes difficulties in the rehabilitation of AMI patients after reperfusion. METHODS: In this prospective, multicenter, randomized, double-blind, double-dummy, placebo-controlled trial, we will assigned 673 eligible patients with AMI after reperfusion into 4 groups: receiving Nao-Xin-Tong capsule (NXT), Bu-Yang-Huan-Wu (BYHW) granule (BYHW), Yang-Yin-Tong-Nao granule (YYTN), or placebo. The course of treatment will be 3 months. The primary outcome is HF incidence within 180 days. Nao-Xin-Tong capsule, BYHW granule, and Yang-Yin-Tong-Nao granule are different traditional Chinese medicines used for tonifying Qi and activating blood (TQAB). RESULTS: Three months of TQAB combined with Western medicine may reduce the incidence of HF after reperfusion of AMI and improve patients' quality of life. DISCUSSION: This study will provide an important basis for the application of traditional Chinese medicine in patients with AMI after reperfusion and provide an evidence-based basis for the prevention and treatment strategy of HF after AMI.

Study of extravisual resting-state networks in pituitary adenoma patients with vision restoration.

BACKGROUND: Pituitary adenoma (PA) may compress the optic apparatus, resulting in impaired vision. Some patients can experience improved vision rapidly after surgery. During the early period after surgery, however, the change in neurofunction in the extravisual cortex and higher cognitive cortex has yet to be explored. OBJECTIVE: Our study focused on the changes in the extravisual resting-state networks in patients with PA after vision restoration. METHODS: We recruited 14 patients with PA who experienced visual improvement after surgery. The functional connectivity (FC) of 6 seeds [auditory cortex (A1), Broca's area, posterior cingulate cortex (PCC) for the default mode network (DMN), right caudal anterior cingulate cortex for the salience network (SN) and left dorsolateral prefrontal cortex for the executive control network (ECN)] were evaluated. A paired t test was conducted to identify the differences between two groups of patients. RESULTS: Compared with their preoperative counterparts, patients with PA with improved vision exhibited decreased FC with the right A1 in the left insula lobule, right middle temporal gyrus and left postcentral gyrus and increased FC in the right paracentral lobule; decreased FC with the Broca in the left middle temporal gyrus and increased FC in the left insula lobule and right thalamus; decreased FC with the DMN in the right declive and right precuneus; increased FC in right Brodmann area 17, the left cuneus and the right posterior cingulate; decreased FC with the ECN in the right posterior cingulate, right angular and right precuneus; decreased FC with the SN in the right middle temporal gyrus, right hippocampus, and right precuneus; and increased FC in the right fusiform gyrus, the left lingual gyrus and right Brodmann area 19. CONCLUSIONS: Vision restoration may cause a response of cross-modal plasticity and multisensory systems related to A1 and the Broca. The DMN and SN may be involved in top-down control of the subareas within the visual cortex. The precuneus may be involved in the DMN, ECN and SN simultaneously.

Electronic and optical properties of a novel two-dimensional semiconductor material TlPt2S3: a first-principles study.

Two-dimensional (2D) materials have attracted widespread attention due to their unique physical and chemical properties. Here, by using density functional theory calculations, we suggest a novel 2D TlPt2S3 material whose layered bulk counterpart was synthesized in 1973. Theoretical calculation results indicate that the exfoliating energy of monolayer and bilayer TlPt2S3 is 34.96 meV Å-2 and 36.03 meV Å-2. We systematically studied the electronic and optical properties of monolayer and bilayer TlPt2S3, and revealed that they are indirect band gap semiconductors with band gaps of 2.26 eV and 2.10 eV, respectively. Monolayer and bilayer TlPt2S3 exhibit superior carrier mobility (901.63 cm2 V-1 s-1 and 13635.04 cm2 V-1 s-1 for electron mobility of the monolayer and bilayer, respectively) and photocatalytic performance (as high as 1 × 105 light absorption coefficient in the visible light region). Interestingly, we find that monolayer TlPt2S3 has significant hydrogen evolution performance, while in the bilayer, the electron band distribution shows complete oxygen evolution ability, which indicates that the proposed monolayer and bilayer TlPt2S3 are potential novel 2D materials suitable for photocatalytic water splitting driven by visible light.

Mulberry (Morus alba L.) leaf polysaccharide ameliorates insulin resistance- and adipose deposition-associated gut microbiota and lipid metabolites in high-fat diet-induced obese mice.

Dietary supplements are currently being used to ameliorate metabolic alterations via maintaining gut microflora balance. Mulberry leaf is known as medicine homologous food for its glucose- and lipid-modulating properties. However, the effects of mulberry leaf polysaccharide (MP) on metabolic dysbiosis and gut microbiota are still poorly understood. After extraction and characterization, the beneficial effects of water-soluble MP were evaluated in high-fat diet-induced obese mice. MP treatment could reduce adipose tissue, improve insulin resistance, and alleviate the pathological lesions in colon. Investigation of the underlying mechanism showed that MP modulated gut microbial community by 16S rRNA analysis and reversed the elevation of lipid indexes by plasma lipidomics analysis. Correlation analysis indicated that the abundance of seven key bacterial species and six lipids were closely associated with the metabolic traits, respectively. Overall, MP could ameliorate metabolic disorders, and modify the gut microbiota and lipids. This would greatly facilitate the utilization of MP as a functional food.

16S rRNA genes- and metagenome-based confirmation of syntrophic butyrate-oxidizing methanogenesis enriched in high butyrate loading.

The understanding and enrichment of consortia formed by syntrophic butyrate-oxidizing bacteria and methanogens in the complex environment are crucial for effectively degrading butyrate and preventing acid inhibition. In this study, the better butyrate-tolerated and highly efficient microbial consortia were domesticated and enriched through adding butyric acid ranging from 0.2 to 4.4 g/(L·d). The volumetric biogas production continuously increased to 1.65 L/(L·d). Microbial community diversity showed that a dramatic shift of bacterial structure occurred at BAL of 1.6 g/(L·d) and the structure presented better stability at high BAL. The syntrophic consortia and the main metabolic pathways were revealed through combination of the 16S rDNA and metagenome sequencing analyses. Syntrophomonas was the major butyrate-oxidizing bacterium and oxidized butyrate mainly through ß-oxidaiton. Synergistaceae and Mesotoga acted as the main acetate-oxidizing bacteria. IHT and methanogenesis pathways were strongly enhanced by DMER64 and Methanosarcina as the main H2 carrier and dominant methanogen, respectively.

Three-Dimensional Semantic Segmentation of Pituitary Adenomas Based on the Deep Learning Framework-nnU-Net: A Clinical Perspective.

This study developed and evaluated nnU-Net models for three-dimensional semantic segmentation of pituitary adenomas (PAs) from contrast-enhanced T1 (T1ce) images, with aims to train a deep learning-based model cost-effectively and apply it to clinical practice. METHODS: This study was conducted in two phases. In phase one, two models were trained with nnUNet using distinct PA datasets. Model 1 was trained with 208 PAs in total, and model 2 was trained with 109 primary nonfunctional pituitary adenomas (NFPA). In phase two, the performances of the two models were investigated according to the Dice similarity coefficient (DSC) in the leave-out test dataset. RESULTS: Both models performed well (DSC > 0.8) for PAs with volumes > 1000 mm3, but unsatisfactorily (DSC < 0.5) for PAs < 1000 mm3. CONCLUSIONS: Both nnU-Net models showed good segmentation performance for PAs > 1000 mm3 (75% of the dataset) and limited performance for PAs < 1000 mm3 (25% of the dataset). Model 2 trained with fewer samples was more cost-effective. We propose to combine the use of model-based segmentation for PA > 1000 mm3 and manual segmentation for PA < 1000 mm3 in clinical practice at the current stage.

Computational Screening of High Activity and Selectivity TM/g-C3N4 Single-Atom Catalysts for Electrocatalytic Reduction of Nitrates to Ammonia.

Electrocatalytic reduction of nitrates (NO3RR) selectively generating ammonia (NH3) opens up a new idea for treating nitrates in wastewater, which not only reduces nitrates but also obtains the valuable product ammonia. By first-principles calculations, we explore the activity and selectivity for NO3RR to NH3 of TM/g-C3N4 single-atom catalysts. Six TM/g-C3N4 catalysts (TM = Ti, Os, Ru, Cr, Mn, and Pt) are selected by a four-step screening method. Ru/g-C3N4 is the most promising of these six TM/g-C3N4 catalysts because of its lowest energy barrier and extraordinary selectivity. The origin of the NO3RR activity of Ru/g-C3N4 is explained from the viewpoint of NO3- adsorption. In addition, the hydrogen evolution reaction has also been implied to be uncompetitive for the poor adsorption on H atoms. This work provides a screening mechanism for finding new catalysts for NO3RR to NH3, promotes the development of NO3RR, and provides a stimulating impetus for further experimental exploration.

Sanye Tablet Ameliorates Insulin Resistance and Dysregulated Lipid Metabolism in High-Fat Diet-Induced Obese Mice.

Sanye Tablet (SYT) is a patent prescription widely used in treating T2D and pre-diabetes, especially T2D comorbid with hypertriglyceridemia, for many years in China. However, the underlying mechanism that accounts for the anti-diabetic potential of SYT by regulating lipid-related intermediates remains to be elucidated. This study aimed to investigate the mechanism of SYT on lipid metabolism and insulin sensitivity in high-fat diet (HFD)-induced obese mice by means of combining lipidomics and proteomics. The obese mice models were developed via HFD feeding for 20 consecutive weeks. Mice in the treatment group were given metformin and SYT respectively, and the effects of SYT on body weight, blood glucose, insulin sensitivity, fat accumulation in the organs, and pathological changes in the liver were monitored. Lipid metabolism was examined by lipidomics. Further determination of signaling pathways was detected by proteomics. The biological contributions of the compounds detected in SYT's chemical fingerprint were predicted by network pharmacology. SYT treatment reduced body weight, inhibited viscera and hepatic steatosis lipid accumulation, and prevented insulin resistance. Furthermore, it was found that circulatory inflammatory cytokines were reduced by SYT treatment. In addition, lipidomics analysis indicated that SYT targets lipid intermediates, including diacylglycerol (DAG) and Ceramide (Cer). Mechanistically, SYT positively affected these lipid intermediates by suppressing liver lipogenesis via downregulation of SREBP1/ACC and the JAK/STAT signaling pathway. Our results predicted that astragalin and rosmarinic acid might regulate the JAK-STAT pathway by targeting PIM2 and STAT1, respectively, while paeoniflorin and rosmarinic acid were likely to regulate inflammatory responses by targeting TNFα, IL-6, and IL-4 during T2D. Overall, our study provides supportive evidence for the mechanism of SYT's therapeutic effect on dysregulated lipid metabolism in diabesity.

The synergistic effect of rumen cellulolytic bacteria and activated carbon on thermophilic digestion of cornstalk.

To explore the bioaugmentation of rumen cellulolytic bacteria (RCB) and activated carbon (AC) on thermophilic digestion of cornstalk, biochemical methane potential tests were carried out. Adding RCB or AC can improve methane production, while simultaneous existence of AC (10 g/L) and RCB (5%) obtained the best performance. The maximum cellulose degradation rate, methane production rate and methane yield were 66.92%, 32.2 L/(kgVS·d), and 144.9 L/kgVS, which increased by 30.23%, 51.17%, and 20.35% compared with control group. The cellulolytic and fermentative bacteria (Hydrogenispora), syntrophic acetate-oxidizing bacteria (norank_o_MBA03), and hydrogenotrophic Methanothermobacter were crucial for thermophilic digestion of cornstalk. The enhancement of AC was due to the enrichment of Hydrogenispora and Methanothermobacter, while RCB can increase the abundance of cellulolytic bacteria (Halocella and norank_o_M55-D21) and mixotrophic Methanosarcina. The synergetic effect of AC and RCB owing to the enriched cellulolytic bacteria, the enhanced syntrophic acetate oxidation and the concentrated carbon metabolic flow to methane.