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N -glycosylation has been revealed to be tightly associated with cancer metastasis. As a key transferase that catalyzes the formation of β1,4 N -acetylglucosamine (β1,4GlcNAc) branches on the mannose core of N -glycans, N -acetylglucosaminyltransferase IVa (GnT-IVa) has been reported to be involved in hepatocellular carcinoma (HCC) metastasis by forming N -glycans; however, the underlying mechanisms are largely unknown. In the current study, we found that GnT-IVa was upregulated in HCC tissues and positively correlated with worse outcomes in HCC patients. We found that GnT-IVa could promote tumor growth in mice; notably, this effect was attenuated after mutating the enzymatic site (D445A) of GnT-IVa, suggesting that GnT-IVa regulated HCC progression by forming β1,4GlcNAc branches. To mechanistically investigate the role of GnT-IVa in HCC, we conducted GSEA and GO functional analysis as well as in vitro experiments. The results showed that GnT-IVa could enhance HCC cell migration, invasion and adhesion ability and increase β1,4GlcNAc branch glycans on integrin β1 (ITGB1), a tumor-associated glycoprotein that is closely involved in cell motility by interacting with vimentin. Interruption of β1,4GlcNAc branch glycan modification on ITGB1 could suppress the interaction of ITGB1 with vimentin and inhibit cell motility. These results revealed that GnT-IVa could promote HCC cell motility by affecting the biological functions of ITGB1 through N -glycosylation. In summary, our results revealed that GnT-IVa is highly expressed in HCC and can form β1,4GlcNAc branches on ITGB1, which are essential for interactions with vimentin to promote HCC cell motility. These findings not only proposed a novel mechanism for GnT-IVa in HCC progression but also revealed the significance of N -glycosylation on ITGB1 during the process, which may provide a novel target for future HCC therapy. • GnT-IVa enhances cell motility and correlates with poor outcomes of HCC. • GnT-IVa increases β1,4GlcNAc branches on ITGB1 to promote cell motility of HCC. • β1,4GlcNAc branches on ITGB1 affect interactions with vimentin in HCC. [ABSTRACT FROM AUTHOR]

Quercetin (Que) has been proved to have various biological activities, including anti-oxidation, anti-inflammation and anti-virus, showing great potential in liver protection. However, its water insolubility leads to low bioavailability. Therefore, the development of a suitable drug delivery fashion is imminent. In recent years, liposomes have been widely used in the fields of drug delivery and gene transfer thanks to the cell membrane like structure, easy surface-modification and high encapsulation efficiency. Herein, we fabricated Que loaded anionic liposomes. Galactosylated chitosan (GC) was simply attached to the surfaces of liposomes through electrostatic adsorption to achieve targeted delivery by binding to asialoglycoprotein receptor (ASGPR). The results showed that Que loaded liposomes modified with GC (GC-Que-Lipo) could enrich the liver in mice through tail vein injection. Liposomes could achieve sustained drug release and GC-Que-Lipo promoted M2 polarization of macrophages. More importantly, it could maintain low content of AST, ALT, ALP and high level of GSH while reducing lipid oxidation, thereby protecting the liver from damage in acute liver injury model. In general, we expect to be able to acquire targeted and efficient delivery of quercetin through a facile approach, thus fulfill the prevention and treatment of liver diseases. • A facile approach of liposomes surface modification for drug targeted delivery system was developed. • The GC-Que-Lipo could enrich the liver of mice via tail vein injection and prevent liver from acute injury. • The encapsulation efficiency of quercetin liposomes reached up to 97.38 %. • The sustained and controlled release of drug loaded liposomes enhanced the bioavailability of quercetin. [ABSTRACT FROM AUTHOR]

Abstract: Compressed sensing (CS) is a revolutionary signal acquisition theory, enabling signal acquisition at a rate that is below the Nyquist sampling rate. However, CS signal reconstruction algorithms are computationally expensive. One of the key computation steps in CS algorithms is to iteratively compute a Cholesky decomposition. Modern application acceleration devices, such as FPGAs and GPUs, can accelerate Cholesky decomposition and CS signal reconstruction computation. This paper presents high performance parallel Cholesky decomposition algorithms for GPU and FPGA implementation. For GPUs, an optimized Cholesky decomposition algorithm is developed with high parallelism, reduced data copying, and improved memory access. For FPGAs, a dedicated pipelined hardware architecture for Cholesky decomposition is designed. Only one pipelined triangular linear equation solver is needed for solving Cholesky decomposition and Cholesky decomposition-based linear equation systems. Moreover, CS signal reconstruction algorithms are accelerated on GPUs and FPGAs for fast signal recovery based on our iterative Cholesky decomposition. Results show that the proposed Cholesky decomposition on FPGAs and GPUs are much faster than LAPACK and MAGMA for small matrices. For accelerating CS signal reconstruction algorithms, our FPGA implementation can achieve around 15× speedup and our GPU implementation can achieve about a 38× speedup compared with the CPU using LAPACK and the hybrid CPU/GPU system with MAGMA. [Copyright &y& Elsevier]

tMCAO, transient middle cerebral artery occlusion; PF11, Pseudoginsenoside-F11; OGD/R, oxygen glucose deprivation/re-oxygenation; NCM, neuron-conditioned media; Jmjd3, Jumonji domain-containing protein 3; CM, conditioned media; LPS, lipopolysaccharide; IFN-γ, interferon-gamma. [Display omitted] • The proportion of M2 microglia/macrophages peaks on day 14 after tMCAO in rats. • PF11 promotes M2 peak advancing to day 3 post-tMCAO in rats. • PF11 increases the expression of anti-inflammatory markers in rats. • PF11 promotes microglia polarization to M2 phenotype in a Jmjd3-dependent manner. • PF11 promotes the protection of M2 microglia on OGD/R-induced neuronal injury. The polarization of microglia/macrophages after cerebral ischemia is critical for post-stroke damage/recovery. Previously, we found that pseudoginsenoside-F11 (PF11), an ocotillol-type saponin, has neuroprotective effects on permanent and transient cerebral ischemia in rats. This study aimed to investigate the effects and potential mechanisms of PF11 on microglia/macrophage polarization following transient cerebral ischemia in rats. In vivo data showed that oral administration of PF11 (12 mg/kg) significantly attenuated cognitive deficits and sensorimotor dysfunction, infarct volume and brain edema in transient middle cerebral artery occlusion (tMCAO)-treated rats, as well as reduced the loss of neurons and the over-activation of microglia in penumbra of ipsilateral striatum and cortex. Notably, the proportion of M2 microglia/macrophages in the total activated microglia/macrophages peaked on day 14 after tMCAO in rats, while PF11 promoted its peak advancing to day 3 post-tMCAO, which allowing the damaged brain to enter the repair period more quickly. Furthermore, PF11 increased the expression of anti-inflammatory markers and decreased the expression of pro-inflammatory markers in ipsilateral striatum and cortex. In addition, in vitro data showed that PF11 inhibited the induction of M1 microglia by oxygen glucose deprivation/re-oxygenation (OGD/R)-induced neurons, and promoted the polarization of microglia to M2 phenotype in a Jumonji domain-containing protein 3 (Jmjd3)-dependent manner. Moreover, PF11 promoted the protection of M2 microglia and attenuated the exacerbation of M1 microglia on OGD/R-induced neuronal damage. Taken together, these results indicate that PF11 protects ischemic neurons by promoting M2 microglia/macrophage polarization in a Jmjd3-dependent manner, ultimately facilitating the functional recovery following transient cerebral ischemia. [ABSTRACT FROM AUTHOR]

Macrophages are a type of functionally plastic cells that can create a pro-/anti-inflammatory microenvironment for organs by producing different kinds of cytokines, chemokines, and growth factors to regulate immunity and inflammatory responses. In addition, they can also be induced to adopt different phenotypes in response to extracellular and intracellular signals, a process defined as M1/M2 polarization. Growing evidence indicates that glycobiology is closely associated with this polarization process. In this research, we review studies of the roles of glycosylation, glucose metabolism, and key lectins in the regulation of macrophages function and polarization to provide a new perspective for immunotherapies for multiple diseases. Schematic representation of glycosylation, glucose metabolism and lectins associated with macrophages polarization. The polarization phenotypes of macrophages are dramatically influenced by alterations of glycosylation, glucose metabolism and lectins. On account that nearly all functional membrane proteins on the surface of macrophages are glycosylated, various glycosylation modifications such as O -GlcNAylation, N -glycosylation and sialylation can significantly affect their active state and functional characteristics. O -GlcNAylation and N -glycosylation exhibit a relatively complicated effects on macrophages polarization mainly through regulating NF-κB pathway. In details, they can activate NF-κB pathways in some disease models so as to induce macrophages into the pro-inflammatory M1 phenotype while in others, they may possibly inhibit NF-κB pathways to induce macrophages into anti-inflammatory M2 phenotype. By comparison, the influence of sialylation on macrophages polarization seems more simple that it can enhance macrophages' tolerance to LPS and primarily induce them into M2 phenotype. Glucose metabolism can provide energy and substrates for glycosylation process, which also contributes to the transition between M1 and M2 type macrophages. The strengthen of glycolysis as well as PPP can induce macrophages into M1 phenotype while enhancement of HBP as well as TCA cycle can induce them into M2 macrophages. Lectins, which can specifically recognize glycans, also contribute to macrophages polarization. Siglec-E，Dectin-1 and Mincle can induce macrophages into M1 phenotype while Siglec-9，Siglec-15 and CD206 can induce macrophages into M2 phenotype. [Display omitted] • This review focuses on the effects of glycobiology for macrophage polarization. • This review explores the consequences of glycosylation, glucose metabolism and lectins in macrophage polarization. • This review proposes new potential immunotherapies targeting for macrophages in the future. [ABSTRACT FROM AUTHOR]

• Ischemic neurons induced neutrophils to polarize into N1 phenotypes. • Ischemic neurons induced macrophages to polarize into M1 phenotypes. • PF11 inhibited the induction of N1/M1 by conditioned media from ischemic neurons. • PF11 promoted the polarization of N2 neutrophils and M2 macrophages. • PF11 alleviated the aggravation of N1/M1 and promoted the protection of N2/M2 on ischemic neurons. Pseudoginsenoside-F11 (PF11), an ocotillol-type saponin, has neuroprotective effects on permanent and transient cerebral ischemia in rats by alleviating autophagic/lysosomal defects and repressing calcium overload, respectively. Ischemic stroke triggers peripheral innate immune cells, mainly neutrophils and macrophages, to infiltrate the damaged brain. The polarization of neutrophils and macrophages after cerebral ischemia is essential for post-stroke damage/recovery. However, it remains elusive whether PF11 ameliorates ischemic neuron injury by regulating the polarization of neutrophils and macrophages. The present study demonstrated for the first time that conditioned media from ischemic neurons induced neutrophils and macrophages to polarize into N1 and M1 phenotypes, respectively. Furthermore, PF11 (30, 100 μM) inhibited the induction of N1 neutrophils by conditioned media from oxygen glucose deprivation/re-oxygenation (OGD/R)-induced ischemic neurons and promoted the polarization of neutrophils to N2 phenotypes. In addition, PF11 (100 μM) attenuated the exacerbation of N1 neutrophils and facilitated the protection of N2 neutrophils on OGD/R-induced neuronal damage. Similarly, PF11 (100 μM) inhibited the induction of M1 macrophages by conditioned media from ischemic neurons and facilitated the polarization of macrophages to M2 phenotypes. What's more, PF11 (100 μM) attenuated the aggravation of M1 macrophages and promoted the protection of M2 macrophages on OGD/R-induced primary neuron injury. In summary, the present study indicates that PF11 ameliorates ischemic neuron damage by regulating neutrophils and macrophages polarization, suggesting that neutrophils and macrophages may be promising targets for the treatment of cerebral ischemia. [ABSTRACT FROM AUTHOR]

• N2 neutrophils ameliorate OGD/R-induced neuronal injury via BDNF/TrkB signaling. • The injury induced by tMCAO exhibit spontaneous recovery over time in rats. • The number of neutrophils in brain is positively correlated with BBB permeability. • The number of neutrophils in brain is positively correlated with CINCs levels. • N2 neutrophils is positively correlated with spontaneous recovery after tMCAO. Neutrophils have been traditionally considered as the major mediators of harmful inflammatory responses in ischemic stroke, whereas accumulating evidence indicates that neutrophils can be polarized into an N2 phenotype. Similar to M2 microglia, N2 neutrophils contribute to resolution of inflammation and may participate in neuroprotection. However, it remains unclear whether N2 neutrophils protect ischemic neurons and whether they are associated with long-term outcomes after transient cerebral ischemia in rats. The present study proved that N2 neutrophils protected against oxygen glucose deprivation/re-oxygenation (OGD/R)-induced primary cortical neuron injury via brain-derived neurotrophic factor/tropomyosin-related kinase B (BDNF/TrkB) signaling. In addition, in vivo studies revealed that transient middle cerebral artery occlusion (tMCAO)-induced injury exhibited spontaneous recovery over time in rats. Moreover, neutrophils could infiltrate the ipsilateral brain parenchyma from the periphery after transient cerebral ischemia. Pearson's correlation analysis indicated that the proportion of N2 neutrophils in ipsilateral brain parenchyma was negatively correlated with the number of degenerating neurons, modified Neurological Severity Score (mNSS), brain water content and infarct volume, and positively correlated with the number of surviving neurons and grip strength. In summary, the present study shows that N2 neutrophils likely participate in spontaneous recovery after transient cerebral ischemia by inhibiting ischemic neuron damage in rats, which indicates that N2 neutrophils may represent promising therapeutic target for promoting recovery after ischemic stroke. [ABSTRACT FROM AUTHOR]

Background: Coronavirus disease 2019 is a type of acute infectious pneumonia and frequently confused with influenza since the initial symptoms. When the virus colonized the patient's mouth, it will cause changes of the oral microenvironment. However, few studies on the alterations of metabolism of the oral microenvironment affected by SARS-CoV-2 infection have been reported. In this study, we explored metabolic alterations of oral microenvironment after SARS-CoV-2 infection. Methods: Untargeted metabolomics (UPLC-MS) was used to investigate the metabolic changes between oral secretion samples of 25 COVID-19 and 30 control participants. To obtain the specific metabolic changes of COVID-19, we selected 25 influenza patients to exclude the metabolic changes caused by the stress response of the immune system to the virus. Multivariate analysis (PCA and PLS-DA plots) and univariate analysis (students' t-test) were used to compare the differences between COVID-19 patients and the controls. Online hiplot tool was used to perform heatmap analysis. Metabolic pathway analysis was conducted by using the MetaboAnalyst 5.0 web application. Results: PLS-DA plots showed significant separation of COVID-19 patients and the controls. A total of 45 differential metabolites between COVID-19 and control group were identified. Among them, 35 metabolites were defined as SARS-CoV-2 specific differential metabolites. Especially, the levels of cis-5,8,11,14,17-eicosapentaenoic acid and hexanoic acid changed dramatically based on the FC values. Pathway enrichment found the most significant pathways were tyrosine-related metabolism. Further, we found 10 differential metabolites caused by the virus indicating the body's metabolism changes after viral stimulation. Moreover, adenine and adenosine were defined as influenza virus-specific differential metabolites. Conclusions: This study revealed that 35 metabolites and tyrosine-related metabolism pathways were significantly changed after SARS-CoV-2 infection. The metabolic alterations of oral microenvironment in COVID-19 provided new insights into its molecular mechanisms for research and prognostic treatment. [ABSTRACT FROM AUTHOR]

The North Qaidam Orogenic Belt (NQOB) is characterized by widespread Palaeozoic magmatism and high to ultrahigh-pressure metamorphism that provides important information regarding the continental crust growth and tectonic evolution in the North Qaidam. Our work focuses on the geology, geochronology, whole-rock geochemistry, and Sr–Nd isotopy of granitoids in the Tanjianshan area in the northern NQOB. The results indicate that the 486–469 Ma adakitic and TTG-like granitoids originated from the partial melting of Middle–Late Mesoproterozoic lower crustal materials with the addition of juvenile mafic lower crust at high pressures under an active continental margin setting. The 396–392 Ma I-type granitoids originated from the partial melting of juvenile crustal materials with the addition of ancient crustal component in a post-collisional setting. Geochronological studies show that there are significant regional differences in magmatism between the northern and rest parts of the NQOB. In the northern part, extensive and prolonged magmatism was widespread during oceanic subduction (~526–460 Ma) and post-collisional extension (~420–350 Ma) but magmatism in the stage of orogenic collision was absent. The Early Palaeozoic adakitic magmatism suggest a significant continental crustal reworking in a subduction zone during 486–469 Ma along the NQOB. The initial oceanic subduction may have occurred prior to ca. 526 Ma. An intense and multiple-stage extension developed at ~420–350 Ma in the northern part of the NQOB following the orogenic collapse. Lithospheric delamination and subsequent mafic magma underplating may be the main geodynamic mechanism resulting in the uprooting of the continental crust and ore mineralization in response. [ABSTRACT FROM AUTHOR]

The widespread Early Cretaceous plutons intruding along the southern Great Xing'an Range (SGXR) provide evidence for tectonic evolution of the region. Petrological, geochemical, zircon U–Pb geochronology, and zircon Hf isotopic studies are conducted on intrusions from Bianjiadayuan and Hongling areas. These suites classify as A2-type granites and monzodiorites, respectively. The 138–133 Ma A2-type granites originated from partial melting of continental crustal materials at high temperatures and shallow depths with significant addition of juvenile mafic lower crust sourced from a metasomatized mantle. The 136–134 Ma monzodiorites originated from the partial melting of an enriched mantle that was modified by melts of a previously subducted slab coupled with crustal contamination. The Early Cretaceous magmatism in the SGXR occurred in two periods: ∼145–136 Ma (peak at ∼139 Ma; εHf (t) = 5 to 10) and ∼136–130 Ma (peak at ∼131 Ma; εHf (t) = –10 to 15). The Early Cretaceous granite–monzodiorite suite in the SGXR suggests a bimodal magmatism in an extensional setting. The ∼145–130 Ma magmatism may have been triggered by asthenospheric upwelling induced by the Mongol–Okhotsk oceanic slab breakoff and large-scale lithospheric delamination resulting from post-orogenic extension. The variation of subduction direction of the Paleo-Pacific Ocean likely triggered a change in stress regime at ca. 136 Ma and likely promoted the lithospheric delamination beneath the SGXR resulting in intense magmatism originating from various sources. As such, the Paleo-Pacific Oceanic subduction likely played an important role in the Early Cretaceous magmatism in the SGXR. [ABSTRACT FROM AUTHOR]

The glycosylation levels of proteins in cancer cells are closely related to cancer invasion and migration. CD44 is a transmembrane glycoprotein that is significantly overexpressed in a variety of tumor cells and has been proven to promote the migration and motility of cancer cells, but the effect of its N-glycosylation modification on CD44 protein function in tumors is less studied. Here, we investigated the effect of six N-glycan chains (N25/57/100/110/120/255) on CD44s localization, function and stability in hepatocarcinoma cells. When the six sites were mutated, we found that CD44s lost its membrane localization in Huh7 and MHCC-97H cells. On this basis, we identified three glycosylation sites on CD44s (N57, N100 and N110) that played key roles in intracellular localization. When N57, N100 and N110 were mutated together, CD44 localized to the cytoplasm, while another three-site mutant (N25/N120/N255) was still anchored to the membrane. In addition, the ability of CD44-N57Q/N100Q/N110Q to promote the metastasis and invasion of Huh7 and 97H cells was weakened compared with that of CD44-N25Q/N120Q/N255Q. Furthermore, CD44-N57Q/N100Q/N110Q accumulated abnormally in the ER, and a high level of the ER stress (ERS) marker BiP was detected at the same time compared with wild-type CD44. When the lysosome inhibitor CQ was added, the content of mutant protein that triggered ERS significantly increased, which indicated that the degradation mode of CD44-N57Q/N100Q/N110Q after ERS was mainly through the lysosomal pathway (ERLAD). The results revealed that the N-glycosylation sites N57, N100 and N110 mutated on CD44s affected its function and degraded it by lysosomes after triggering ERS. These findings provide data for new studies on ER-related degradation, further promote the study of the glycan chain function of CD44 and furnish new ideas for the treatment of liver cancer metastasis. • N57, N100 and N110 are crucial N-glycosylation sites for CD44 function of promoting metastasis in HCC. • The lack of glycan chains on N57, N100 and N110 leads to translocation from membrane to cytoplasm of CD44 in HCC cells. • CD44-N57Q/N100Q/N110Q is degraded through ERLAD after triggering ER stress. [ABSTRACT FROM AUTHOR]

• PF11 enhances the OGD-induced promotion of myelin debris phagocytosis by microglia. • PF11 decreases the release of OGD-induced microglial pro-inflammatory factor. • PF11 promotes microglial phagocytosis of myelin debris through CR3. • PF11 attenuates cerebral ischemic injury through CR3. After ischemic stroke, the degenerated myelin caused by ischemic injury cannot be rapidly cleared away by microglia and interferes with the recovery process. Complement receptor 3 (CR3, CD11b/CD18), belonging to β 2 integrin family primarily expressed in phagocytes, is involved in the microglial phagocytosis of myelin debris. We previously found that pseudoginsenoside-F11 (PF11), an ocotillol-type saponin, exerts neuroprotective effects against ischemic stroke and neuroinflammation. In the present study, we investigated the promotion of PF11 on oxygen-glucose deprivation (OGD)-induced microglial phagocytosis of myelin debris, the neuroprotection of PF11 on permanent middle cerebral artery occlusion (pMCAO)-induced ischemic stroke, and the possible role of CR3. The results indicated that PF11 (50 μM) accelerated the OGD-induced promotion of myelin debris phagocytosis by microglia in the early stage of OGD (2 h, 4 h, 8 h), which was significantly inhibited by anti-CD11b mAb or down-regulated by CD11b-specific siRNA. Meanwhile, PF11 strengthened the OGD-activated RhoA/ROCK signaling associated with the internalization during myelin debris phagocytosis through CR3. Consistently, the anti-CD11b mAb could markedly attenuated the nrueoprotective effects of PF11 (12 mg/kg, i.v.) on infarction and brain edema, neurological functions and loss of neurons of pMCAO rats. These findings suggest that PF11 accelerates the phagocytosis of myelin debris by microglia mainly through CR3, which may likely contribute to its neuroprotection against ischemic stroke. [ABSTRACT FROM AUTHOR]

Pseudoginsenoside-F11 (PF11), an ocotillol-type saponin, has been reported to have anti-inflammatory properties, but the effects of PF11 on acute lung inflammation were unknown. The present study aimed to investigate the protective effects and potential mechanisms of PF11 on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in male BALB/c mice. After being treated with PF11 (3, 10, and 30 mg/kg, intravenous) once a day for 3 consecutive days, the mice were challenged by intratracheal instillation of LPS, and then their lung tissues and bronchoalveolar lavage fluid (BALF) were collected for further analysis. The results showed that PF11 attenuated LPS-induced ALI, with alleviated histopathological damage, decreased lung wet/dry weight ratio, and reduced protein concentration and inflammatory cells number in BALF. Moreover, PF11 reversed the LPS-induced increases of mRNA expression and protein levels of interleukin-6, tumor necrosis factor-α, and interleukin-1β. Meanwhile, PF11 decreased LPS-induced myeloperoxidase activity and neutrophil infiltration in lung tissue by reducing the expression of macrophage inflammatory protein-2 and intercellular adhesion molecule-1, as well as enhanced neutrophil clearance by accelerating neutrophils apoptosis and their phagocytosis by alveolar macrophages. In conclusion, these results indicated that PF11 significantly attenuated LPS-induced ALI through suppressing neutrophil infiltration and accelerating neutrophil clearance, suggesting its potential in the treatment of ALI. [ABSTRACT FROM AUTHOR]

[Display omitted] • Endothelial progenitor cells (EPCs) were an emerging stem cell therapy candidate. • The extracellular vesicles (EVs) released from EPCs have pro-angiogenesis power. • TFEB engineering can improve the anti-ischemia activity of EPC-EVs. • Thermal-/pH-responsive hydrogel is suitable for critical limb ischemia therapy. Critical limb ischemia (CLI) is the terminal stage of peripheral arterial disease (PAD), always leading to a high risk of amputations and deaths. While the majority of strategies only focus on recanalization of patients, these methods are unsatisfactory due to limited therapeutic effect and surgical damage. Herein, a multifunctional engineered-extracellular vesicles (EVs)/hydrogel system is designed for facilitating angiogenesis and protecting the ischemic muscle tissue via a non-surgical delivery strategy. The EVs released from endothelial progenitor cells (EPCs), an emerging stem cell therapy candidate, were used to play the pro-angiogenesis role. Transcription factor EB (TFEB), a master regulator of autophagy, was utilized to genetically modify the EVs for building TFEB-engineered EVs (TFEB-EVs) to reduce the impairment of muscle. Moreover, a thermal-/pH-responsive hydrogel (FPD), prepared by F127, polyethylenimine (PEI), and oxidized dextran, served as a sensitive controlled-release carrier and improved the stability of TFEB-EVs in vivo , simultaneously. Our results exhibited that TFEB-EVs/FPD significantly strengthened neovascularization, alleviated muscle damage, and protected the limb function of CLI mice. Overall, TFEB-EVs/FPD offered a convenient, safe, and non-surgical approach for rescuing CLI. [ABSTRACT FROM AUTHOR]