Perfluorooctanoic acid effect and microbial mechanism to methane production in anaerobic digestion.

Perfluorooctanoic acid (PFOA) as emerging pollutants was largely produced and stable in nature environment. Its fate and effect to the wasted sludge digestion process and corresponding microbial mechanism was rarely reported. This study investigated the different dose of PFOA to the wasted sludge digestion process, where the methane yield and microbial mechanism was illustrated. The PFOA added before digestion were 0-10000 µg/L, no significant variation in daily and accumulated methane production between each group. The 9th day methane yield was significantly higher than other days (p < 0.05). The soluble protein was significantly decreased after 76 days digestion (p < 0.001). The total PFOA in sludge (R2 = 0.8817) and liquid (R2 = 0.9083) phase after digestion was exponentially correlated with PFOA dosed. The PFOA in liquid phase was occupied 54.10 ± 18.38% of the total PFOA in all reactors. The dewatering rate was keep decreasing with the increase of PFOA added (R2 = 0.7748, p < 0.001). The mcrA abundance was significantly correlated with the pH value and organic matter concentration in the reactors. Chloroflexi was the predominant phyla, Aminicenantales, Bellilinea and Candidatus_Cloacimonas were predominant genera in all reactors. Candidatus_Methanofastidiosum and Methanolinea were predominant archaea in all reactors. The function prediction by FAPROTAX and Tax4fun implied that various PFOA dosage resulted in significant function variation. The fermentation and anaerobic chemoheterotrophy function were improved with the PFOA dose. Co-occurrence network implied the potent cooperation among the organic matter degradation and methanogenic microbe in the digestion system. PFOA has little impact to the methane generation while affect the microbe function significantly, its remaining in the digested sludge should be concerned to reduce its potential environmental risks.

Comparison and optimization of different CRISPR/Cas9 donor-adapting systems for gene editing.

Gene knock-in in mammalian cells usually uses homology-directed repair (HDR) mechanism to integrate exogenous DNA template into the target genome site. However, HDR efficiency is often low, and the co-localization of exogenous DNA template and target genome site is one of the key limiting factors. To improve the efficiency of HDR mediated by CRISPR/Cas9 system, our team and previous studies fused different adaptor proteins with SpCas9 protein and expressed them. By using their characteristics of binding to specific DNA sequences, many different CRISPR/SpCas9 donor adapter gene editing systems were constructed. In this study, we used them to knock-in eGFP gene at the 3'-end of the terminal exon of GAPDH and ACTB genes in HEK293T cells to facilitate a comparison and optimization of these systems. We utilized an optimized donor DNA template design method, validated the knock-in accuracy via PCR and Sanger sequencing, and assessed the efficiency using flow cytometry. The results showed that the fusion of yGal4BD, hGal4BD, hLacI, hTHAP11 as well as N57 and other adaptor proteins with the C-terminus of SpCas9 protein had no significant effect on its activity. At the GAPDH site, the donor adapter systems of SpCas9 fused with yGal4BD, hGal4BD, hLacI and hTHAP11 significantly improved the knock-in efficiency. At the ACTB site, SpCas9 fused with yGal4BD and hGal4BD significantly improved the knock-in efficiency. Furthermore, increasing the number of BS in the donor DNA template was beneficial to enhance the knock-in efficiency mediated by SpCas9-hTHAP11 system. In conclusion, this study compares and optimizes multiple CRISPR/Cas9 donor adapter gene editing systems, providing valuable insights for future gene editing applications.

The Effects of Bone Cement Volume in Percutaneous Vertebroplasty for Thoracolumbar Junction Vertebral Compression Fractures: A Clinical Comparative Study.

We compared the outcomes of patients treated with different volumes of polymethyl methacrylate bone cement during percutaneous vertebroplasty (PVP) for thoracolumbar vertebral compression fractures. We performed a comparative, retrospective study of 316 patients who underwent PVP for a single-level thoracolumbar vertebral compression fracture. Patients were divided into two groups: group A (≤5 mL; n = 146) and group B (>5 mL; n = 170). The visual analogue scale (VAS) for pain and the Roland-Morris Disability Questionnaire (RDQ) scores were compared between the two groups at 1 week and at 1, 6, 12, and 24 months after PVP. The incidence of cement leakage into the intervertebral discs was evaluated by a postoperative lateral radiograph assessment. Patients were evaluated for new fractures 1 and 2 years after PVP or when new fractures were suspected. Among the 316 patients enrolled, 245 completed the clinical research. No difference between groups A and B in terms of the VAS, RDQ, and rate of complications at all time points after surgery was observed. The presence of intervertebral disc leakage was a relative risk (RR) for subsequent total vertebral fracture (RR, 6.42; 95% confidence interval (CI), 2.72-14.19; P < 0.0001) and adjacent vertebral fracture (RR, 8.03; 95% CI, 2.74-23.54; P = 0.0001). A high volume of bone cement may increase the rate of subsequent total and adjacent vertebral fractures. However, the occurrence of intervertebral disc leakage is the principal risk factor for these negative outcomes of PVP.

Integrating Microbial Protein Production and Harvest Systems into Pilot-Scale Recirculating Aquaculture Systems for Sustainable Resource Recovery: Linking Nitrogen Recovery to Microbial Communities.

In aquaculture, it is important to raise the nitrogen recovery efficiency (NRE) to improve sustainability. To achieve this, recovery of microbial protein (RMP), instead of nitrification/denitrification in conventional wastewater treatment, is a promising approach whose microbiological mechanisms must be characterized. Here, periodic RMP was conducted in an in situ biofloc-based aquaculture system (IBAS) and a separating assimilation reactor-based recirculating aquaculture system (SRAS). Kinetic analysis indicated that a microbial biomass level of 3 g L-1 was optimal for inorganic N removal, and excess biomass was harvested to improve the NRE. Unlike the IBAS, the SRAS eliminated the fluctuation in water quality caused by the RMP. Periodic RMP significantly increased the NRE to 44-57% by promoting the filamentous bacterium Herpetosiphon and suppressing anaerobic denitrifiers. Aerobic chemoheterotrophy was the main microbial metabolic process for energy. After RMP, nitrate reductase-encoded functional genes (napA and narG) significantly decreased, while nitrite reductase-encoded functional genes, especially nirK, significantly increased. Co-occurrence networks analysis indicated that the cooperation and competition among organic matter degraders, filamentous bacteria, nitrifiers, and denitrifiers determined the microbial protein yield. These results provide fundamental insights into the influence of the RMP on microbial communities and functions, which is important for realizing sustainable aquaculture.

Deficiency of Mitochondrial Glycerol 3-Phosphate Dehydrogenase Contributes to Hepatic Steatosis.

Mitochondrial glycerol 3-phosphate dehydrogenase (mGPDH) is an integral component of the respiratory chain, and recent studies have suggested that it plays an important role in hepatic glucose homeostasis. However, its function in hepatic lipid metabolism is unclear. Here, we identified a role for mGPDH in nonalcoholic fatty liver disease (NAFLD). Specifically, mGPDH expression and activity were lower in fatty livers from patients and mice with NAFLD (ob/ob, high-fat diet [HFD] and db/db). Liver-specific depletion of mGPDH in mice or mGPDH knockdown in cultured hepatocytes exacerbated diet-induced triglyceride accumulation and steatosis through enhanced lipogenesis. RNA-sequencing revealed that mGPDH regulated endoplasmic reticulum (ER)-related proteins and processes. mGPDH deletion exacerbated tunicamycin (ER stress inducer)-induced hepatic steatosis, whereas tauroursodeoxycholic acid (ER stress inhibitor) rescued mGPDH depletion-induced steatosis on an HFD. Moreover, ER stress induced by mGPDH depletion could be abrogated by the intracellular Ca2+ chelator 1,2-bis (2-aminophenoxy) ethane N,N,N´,N´-tetraacetic acid acetoxymethyl ester, mitochondrial permeability transition pore (mPTP) inhibitor cyclosporine A, or cyclophilin-D (Cyp-D) knockdown. mGPDH promoting Cyp-D ubiquitination was also observed. Finally, liver-specific mGPDH overexpression attenuated hepatic steatosis in ob/ob and HFD mice. Conclusion: mGPDH is a pivotal regulator of hepatic lipid metabolism. Its deficiency induces ER stress by suppressing Cyp-D ubiquitination, a key regulator of the mitochondrial Ca2+ conductance channel mPTP, and results in hepatic steatosis. mGPDH may be a potential therapeutic target for the treatment of NAFLD.

Hydrodynamic Process in the Langmuir-Blodgett Film Method.

Preparing organic coatings in a very controlled manner through the spreading of organic molecules on the water surface is one of the emphases for research in Langmuir-Blodgett (LB) technology. For preparing a homogeneous film and improving the quality of the film, it is our concern to have a deeper understanding of the dynamic process involved in spreading. Here, we present an overview of the hydrodynamic process under the influence of assisting the spreading solvent, which mainly focuses on the mechanical mechanisms of related phenomena. A typical spreading experiment of two-component mixed droplets on water substrate for the purpose of preparing LB films was carried out in this research. We perform the spreading of a liquid of silicone oil and oleic acid mixture on the horizontal surface of another immiscible deep water substrate, where the volatile silicone oil is the assisting spreading solvent with low viscosity. We find that it needs to exceed a certain critical value (60% in our experiment) to achieve a uniform and centrosymmetric spreading process, which is a key factor for getting a homogeneous film. We observe that the evolution of a large droplet into liquid film and then into small droplets under the action of surface tension gradient in experiments. Gravity-viscous and surface tension-viscous dominate successively in the whole spreading process, with its spreading radius r(t) ∝ t1/4 and r(t) ∝ t3/4, respectively. However, we also obtain singular values of scaling exponents -0.033 and -0.180, which is attributed to nonuniform distribution of the Laplace pressure caused by different curvatures near the capillary wave.

High-throughput screening with nucleosome substrate identifies small-molecule inhibitors of the human histone lysine methyltransferase NSD2.

The histone lysine methyltransferase nuclear receptor-binding SET domain protein 2 (NSD2, also known as WHSC1/MMSET) is an epigenetic modifier and is thought to play a driving role in oncogenesis. Both NSD2 overexpression and point mutations that increase its catalytic activity are associated with several human cancers. Although NSD2 is an attractive therapeutic target, no potent, selective, and bioactive small molecule inhibitors of NSD2 have been reported to date, possibly due to the challenges of developing high-throughput assays for NSD2. Here, to establish a platform for the discovery and development of selective NSD2 inhibitors, we optimized and implemented multiple assays. We performed quantitative high-throughput screening with full-length WT NSD2 and a nucleosome substrate against a diverse collection of bioactive small molecules comprising 16,251 compounds. We further interrogated 174 inhibitory compounds identified in the primary screen with orthogonal and counter assays and with activity assays based on the clinically relevant NSD2 variants E1099K and T1150A. We selected five confirmed inhibitors for follow-up, which included a radiolabeled validation assay, surface plasmon resonance studies, methyltransferase profiling, and histone methylation in cells. We found that all five NSD2 inhibitors bind the catalytic SET domain and one exhibited apparent activity in cells, validating the workflow and providing a template for identifying selective NSD2 inhibitors. In summary, we have established a robust discovery pipeline for identifying potent NSD2 inhibitors from small-molecule libraries.

Pattern Formation in Drying Sessile and Pendant Droplet: Interactions of Gravity Settling, Interface Shrinkage, and Capillary Flow.

We reported the interactions of the gravitational sedimentation, interface shrinkage, and outward capillary flow in drying droplets. This coupling effect is the inference we draw from deposition patterns of both sessile and pendant droplets, which contain particles of different sizes, evaporating on a patterned substrate. The deposition difference between sessile and pendant droplets containing microparticles indicated that gravitational sedimentation has a significant influence on the deposition morphology. The phase diagram shows that the particle deposition process can be divided into two stages: in the first stage, the competition between the interface shrinkage and the gravitational sedimentation determines whether the particles can be captured by the liquid-air interface; in the second stage, the capillary flow takes the particles inside the droplet toward the edge. The deposition morphology is the result of competition and cooperation interactions of the free setting, interface shrinkage, and outward capillary flow.

Tunable Dirac cones in two-dimensional acoustic metamaterials with matryoshka structure.

Dirac cones of an acoustic system are the foundation of most topological phase transitions and topological states and have recently become a research hotspot. Although the Dirac cones, Dirac-like cones, double Dirac cones, and semi-Dirac points are all skillfully designed, it is still indispensable to realize a tunable Dirac cone in a novel acoustic structure. This paper proposes two-dimensional acoustic metamaterials with matryoshka structure to achieve tunable Dirac cones and topological spin states. Dirac points can be obtained on the dispersion curves owing to the high symmetry. The concentric circular scattering units of the matryoshka structure are arranged in honeycomb lattices. By a rotating-scatterer mechanism to break the symmetry, the Dirac cone at K (K') is split and the topological spin states appear at the band valley. The existence of a topological transition with opposite Chern numbers as the rotating angle varies is also verified, and helical edge states are obtained along the interfaces separating the topologically opposite spin states insulators. Moreover, the frequency of the Dirac cone is tuned by rotating the inner structure in a double-layer matryoshka structure.

Acoustic valley edge states in a graphene-like system with sub-wavelength resonator.

Recently, the study of topological phase transitions and edge states for acoustic wave systems has become a research hotspot. However, most current studies on topological edge states are based on Bragg scattering, which is not practical to apply in situations involving low-frequency sound because of the large structural dimensions. Therefore, the authors construct, in this study, a graphene-like structure based on a sub-wavelength resonant unit Helmholtz resonator and adjust the acoustic capacitance diameter of adjacent units to change the local resonance frequency, and thereby impose the degeneracy of the Dirac cone and topological spin states, which is characterized by valley Chern numbers of opposite sign. The authors also check topological valley edge states at zigzag and armchair interfaces and find that gapless topological valley edge states only appear at zigzag interfaces, whereas armchair interfaces host gap edge states. Moreover, the results show that the transmission properties of edge states in a zigzag rectangular waveguide are immune to backscattering and defects.

Drop Capturing Based on Patterned Substrate in Space.

In this work, we introduced a method for capturing aqueous drop based on a patterned substrate in space. Through the manipulation test of a colloidal drop, it could be verified that this patterned substrate had excellent control ability for aqueous drop in microgravity condition. The confinement mechanism of this substrate was clarified, which showed that drops with different volume could be pinned and attracted at a given area on the substrate. The confinement capability was related to the gravity effect, and the patterned substrate could confine aqueous drops with larger volume under microgravity than in normal gravity. With advantages of simple operation and strong capability to control large drops, this technique exhibited the wide application prospect in the fields of fluid management, biosensing, and pharmacy in microgravity condition in the future.

Plasma Asprosin Concentrations Are Increased in Individuals with Glucose Dysregulation and Correlated with Insulin Resistance and First-Phase Insulin Secretion.

BACKGROUND: Adipokines are reported to participate in many common pathologic processes of glucose dysregulation, such as insulin resistance, ß-cell dysfunction, and chronic inflammation. OBJECTIVE: To detect the concentrations of plasma asprosin in subjects with impaired glucose regulation (IGR) and newly diagnosed type 2 diabetes (nT2DM) and its relationship to parameters of glucose and lipid metabolism, insulin resistance, and pancreatic ß-cell function. METHODS: 143 eligible participants were included and were divided into three groups including normal glucose regulation (NGR, n = 52), IGR (n = 40), and nT2DM group (n = 51). The intravenous glucose tolerance test (IVGTT) and clinical and biochemical parameters were measured in all participants. RESULTS: Plasma asprosin levels were higher in IGR (82.40 ± 91.06 ng/mL, P < 0.001) and nT2DM (73.25 ± 91.69 ng/mL, P < 0.001) groups compared with those in the NGR (16.22 ± 9.27 ng/mL) group, especially in IGR subjects. Correlation analysis showed that plasma asprosin levels were positively correlated with waist circumference (Wc), fasting plasma glucose (FPG), postchallenge plasma glucose (2hPG), HbA1c, triglyceride (TG), and homeostasis model assessment for insulin resistance (HOMA-IR) and negatively correlated with homeostasis model assessment for ß-cell function (HOMA-ß), area under the curve of the first-phase (0-10 min) insulin secretion (AUC), acute insulin response (AIR), and glucose disposition index (GDI) (all P < 0.05). Multiple logistical regression analyses revealed that plasma asprosin concentrations were significantly correlated with IGR and nT2DM after controlling for age, sex, BMI, and WHR. CONCLUSIONS: Circulating asprosin might be a predictor of early diagnosis in DM and might be a potential therapeutic target for prediabetes and T2DM.

Evidence of the Role of R-Spondin 1 and Its Receptor Lgr4 in the Transmission of Mechanical Stimuli to Biological Signals for Bone Formation.

The bone can adjust its mass and architecture to mechanical stimuli via a series of molecular cascades, which have been not yet fully elucidated. Emerging evidence indicated that R-spondins (Rspos), a family of secreted agonists of the Wnt/ß-catenin signaling pathway, had important roles in osteoblastic differentiation and bone formation. However, the role of Rspo proteins in mechanical loading-influenced bone metabolism has never been investigated. In this study, we found that Rspo1 was a mechanosensitive protein for bone formation. Continuous cyclic mechanical stretch (CMS) upregulated the expression of Rspo1 in mouse bone marrow mesenchymal stem cells (BMSCs), while the expression of Rspo1 in BMSCs in vivo was downregulated in the bones of a mechanical unloading mouse model (tail suspension (TS)). On the other hand, Rspo1 could promote osteogenesis of BMSCs under CMS through activating the Wnt/ß-catenin signaling pathway and could rescue the bone loss induced by mechanical unloading in the TS mice. Specifically, our results suggested that Rspo1 and its receptor of leucine-rich repeat containing G-protein-coupled receptor 4 (Lgr4) should be a novel molecular signal in the transmission of mechanical stimuli to biological signal in the bone, and this signal should be in the upstream of Wnt/ß-catenin signaling for bone formation. Rspo1/Lgr4 could be a new potential target for the prevention and treatment of disuse osteoporosis in the future.

The role of Arabidopsis Actin-Related Protein 3 in amyloplast sedimentation and polar auxin transport in root gravitropism.

Gravitropism is vital for shaping directional plant growth in response to the forces of gravity. Signals perceived in the gravity-sensing cells can be converted into biochemical signals and transmitted. Sedimentation of amyloplasts in the columella cells triggers asymmetric auxin redistribution in root tips, leading to downward root growth. The actin cytoskeleton is thought to play an important role in root gravitropism, although the molecular mechanism has not been resolved. DISTORTED1 (DIS1) encodes the ARP3 subunit of the Arabidopsis Actin-Related Protein 2/3 (ARP2/3) complex, and the ARP3/DIS1 mutant dis1-1 showed delayed root curvature after gravity stimulation. Microrheological analysis revealed that the high apparent viscosity within dis1-1 central columella cells is closely associated with abnormal movement trajectories of amyloplasts. Analysis using a sensitive auxin input reporter DII-VENUS showed that asymmetric auxin redistribution was reduced in the root tips of dis1-1, and the actin-disrupting drug Latrunculin B increased the asymmetric auxin redistribution. An uptake assay using the membrane-selective dye FM4-64 indicated that endocytosis was decelerated in dis1-1 root epidermal cells. Treatment and wash-out with Brefeldin A, which inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus, showed that cycling of the auxin-transporter PIN-FORMED (PIN) proteins to the plasma membrane was also suppressed in dis1-1 roots. The results reveal that ARP3/DIS1 acts in root gravitropism by affecting amyloplast sedimentation and PIN-mediated polar auxin transport through regulation of PIN protein trafficking.

Protein kinase profiling assays: a technology review.

Protein kinases have become one of the most intensively pursued classes of drug targets for many diseases such as cancers and inflammatory diseases. Kinase profiling work seeks to understand general selectivity trends of lead compounds across the kinome, which help with target selection, compound prioritization, and potential implications in toxicity. Under the current drug discovery process, screening of compounds against comprehensive panels of kinases and their mutants has become the standard approach. Many screening assays and technologies which are compatible for high-throughput screening (HTS) against kinases have been extensively pursued and developed.

[Preparation of elastic porous cell scaffold fabricated with combined polydimethylsiloxane (PDMS) and hydroxyapatite (HA)].

Polydimethylsiloxane (PDMS) and hydroxyapatite (HA) were combined in our laboratory to fabricate an elastic porous cell scaffold with pore-forming agent, and then the scaffold was used as culture media for rat bone marrow derived mesenchymal stem cells (rBMSCs). Different porous materials (square and circular in shape) were prepared by different pore-forming agents (NaCl or paraffin spheres) with adjustable porosity (62%-76%). The HA crystals grew on the wall of hole when the material was exposed to SBF solutions, showing its biocompatibility and ability to support the cells to attach on the materials.

Evaporative self-assembly in colloidal droplets: Emergence of ordered structures from complex fluids.

Colloidal droplet evaporation is an intriguing and intricate phenomenon that has captured the interest of scientists across diverse disciplines, including physical chemistry, fluid dynamics, and soft matter science, over the past two decades. Despite being a non-equilibrium system with inherent challenges posed by coffee ring formation and Marangoni effects, which hinder the precise control of deposition patterns, evaporative self-assembly presents a convenient and cost-effective approach for generating arrays of well-ordered structures and functional patterns with wide-ranging applications in inkjet printing, photonic crystals, and biochemical assays. In the realm of printed electronics and photonics, effectively mitigating coffee rings while achieving uniformity and orderliness has emerged as a critical factor in realising the next generation of large-area, low-cost, flexible devices that are exceptionally sensitive and high-performance. This review highlights the evaporative self-assembly process in colloidal droplets with a focus on the intricate mechanical environment, self-assembly at diverse interfaces, and potential applications of these assembling ordered structures.

Plasma Asprosin Concentrations are Associated with Progression of Diabetic Kidney Disease.

Purpose: To explore the expression of asprosin in subjects with pre-DKD and DKD and to analyze its relationship with kidney injury, inflammation, and glucose and lipid metabolism. Methods: Based on urine albumin:creatinine ratio (UACr), participants were divided into DM, pre-DKD, and DKD groups. Relevant human physiological and biochemical parameters were detected in the three groups. Results: We found relatively higher levels of asprosin in both pre-DKD and DKD groups than the DM group. Moreover, data from the Nephroseq database support increased gene expression of asprosin in kidney tissue from DKD patients. Further correlation analysis revealed that the plasma asprosin level was positively correlated with age, waist circumference, waist:hip ratio, systolic blood pressure, creatinine, UACr, triglycerides, HDL-c, fasting insulin, HOMA-IR, and the inflammatory marker G3P and negatively associated with eGFR. Multiple logistical regression analysis showed that asprosin concentration was significantly associated with pre-DKD and DKD after adjusting for sex, age, BMI, WHR, and HOMA-IR, while this correlation was lost after controlling for G3P. Conclusion: Plasma asprosin is associated with kidney injury in diabetic conditions, and this association might be connected through inflammatory response. Further studies are needed to assess the role and mechanism of asprosin in DKD.

Increasing fish production in recirculating aquaculture system by integrating a biofloc-worm reactor for protein recovery.

Aquaculture, producing half of global fish production, offers a high-quality protein source for humans. Improving nitrogen use efficiency (NUE) through microbial protein recovery is crucial for increasing fish production and reducing environmental footprint. However, the poor palatability and high moisture content of microbial protein make its utilization challenging. Here, a biofloc-worm reactor was integrated into a recirculating aquaculture system (BW_RAS) for the first time to convert microbial protein into Tubificidae (Oligochaeta) biomass, which was used as direct feed for culturing fish. Batch experiments indicated that an aeration rate of 0.132 m3 L -1 h -1 and a worm density of 0.3 g cm-2 on the carrier were optimal for microbial biomass growth and worm predation, respectively. Compared to the biofloc reactor-based recirculating aquaculture system (B_RAS), the BW_RAS improved water quality, NUE, and fish production by 17.1 % during a 120-day aquaculture period. The abundance of heterotrophic aerobic denitrifier Deinococcus in BW_RAS was one order of magnitude higher than in B_RAS, while heterotrophic bacteria Mycobacterium was more abundant in B_RAS. Denitrifiers cooperated with organic matter degraders and nitrogen assimilation bacteria for protein recovery and gaseous nitrogen loss while competing with predatory bacteria. Function prediction and qPCR indicated greater aerobic respiration, nitrate assimilation, nitrification (AOB-amoA), and denitrification (napA, nirK, nirS, nosZI), but lower fermentation in BWR compared to BR. This study demonstrated that BW_RAS increased microbial protein production and aerobic nitrogen cycling through ongoing worm predation, further enhancing fish production to a commercially viable level.

CD9 Counteracts Liver Steatosis and Mediates GCGR Agonist Hepatic Effects.

Glucagon receptor (GCGR) agonism offers potentially greater effects on the mitigation of hepatic steatosis. However, its underlying mechanism is not fully understood. Here, it screened tetraspanin CD9 might medicate hepatic effects of GCGR agonist. CD9 is decreased in the fatty livers of patients and upregulated upon GCGR activation. Deficiency of CD9 in the liver exacerbated diet-induced hepatic steatosis via complement factor D (CFD) regulated fatty acid metabolism. Specifically, CD9 modulated hepatic fatty acid synthesis and oxidation genes through regulating CFD expression via the ubiquitination-proteasomal degradation of FLI1. In addition, CD9 influenced body weight by modulating lipogenesis and thermogenesis of adipose tissue through CFD. Moreover, CD9 reinforcement in the liver alleviated hepatic steatosis, and blockage of CD9 abolished the remission of hepatic steatosis induced by cotadutide treatment. Thus, CD9 medicates the hepatic beneficial effects of GCGR signaling, and may server as a promising therapeutic target for hepatic steatosis.