Discovery of 5-(1-benzyl-1H-imidazol-4-yl)-1,2,4-oxadiazole derivatives as novel RIPK1 inhibitors via structure-based virtual screening.

RIPK1 plays a key role in necroptosis and is associated with various inflammatory diseases. Using structure-based virtual screening, a novel hit with 5-(1-benzyl-1H-imidazol-4-yl)-1,2,4-oxadiazole scaffold was identified as an RIPK1 inhibitor with an IC50 value of 1.3 µM. Further structure-activity relationship study was performed based on similarity research and biological evaluation. The molecular dynamics simulation of compound 2 with RIPK1 indicated that it may act as a type II kinase inhibitor. This study provides a highly efficient way to discover novel scaffold RIPK1 inhibitors for further development.

An injectable, activated neutrophil-derived exosome mimetics/extracellular matrix hybrid hydrogel with antibacterial activity and wound healing promotion effect for diabetic wound therapy.

Chronic diabetic wounds are primarily caused by infection, inflammation, and angiogenesis-related disorders. An ideal approach for treating chronic diabetic wounds is by combining anti-infection strategies, immune microenvironment regulation, and angiogenesis promotion. Vascular endothelial growth factor (VEGF) can promote the proliferation and migration of vascular endothelial cells, thereby promoting angiogenesis. However, the low stability and inability to target lesions limit its application. Polymorphonuclear neutrophil-derived exosomes (PMNExo) exhibit good delivery properties and can be used for the therapeutic delivery of VEGF. Furthermore, they retain the antibacterial ability of polymorphonuclear neutrophils (PMNs). Nonetheless, low PMNExo generation impedes its therapeutic applications. In this study, we prepared exosome mimetics (EM) from PMNs using the extrusion process; as a result, exosome yield significantly improved. To increase the residence of exosomes, an extracellular matrix (ECM) hydrogel, a thermosensitive material that can function as an in situ gel in vivo, was used as an exosome carrier. The active peptides in the ECM regulated the immune microenvironment of the wound. In summary, we loaded ECM with VEGF-encapsulated activated neutrophil exosome mimetics (aPMNEM) to develop VEGF-aPMNEM-ECM hybrid hydrogel for treating chronic wounds. The hydrogel accelerates the regeneration of chronic diabetic wounds. Our study provides a prospective therapy platform involving cytokines for treating different diseases.

Calcium sensing receptor mediated the excessive generation of ß-amyloid peptide induced by hypoxia in vivo and in vitro.

Hypoxia played an important role in the pathogenesis of AD. Hypoxia increased Aß formation, then caused Alzheimer's disease. Calcium sensing receptor (CaSR) was involved in the regulation of cell growth, differentiation, hormonal secretion and other physiological function. Increasing evidence supported CaSR might play a more prominent role in susceptibility to AD, but the role of CaSR in Aß overproduction induced by hypoxia and its mechanisms remain unclear. To investigate whether CaSR mediated the overproduction of Aß induced by hypoxia, immunoblot and immunochemistry were employed to determine the expression of CaSR and BACE1 in hippocampal neurons and tissue and Ca(2+) image system was used to measure [Ca(2+)]i in hippocampal neurons. The content of Aß was detected with ELISA kits. Our research found that hypoxia increased the expression of CaSR in hippocampal neurons and tissue and [Ca(2+)]i in hippocampal neurons. Calhex 231, a selective blocher of CaSR, inhibited the increase in [Ca(2+)]i induced by hypoxia. Hypoxia or GdCl3, an agonist of CaSR, increased the expression of BACE1 in hippocampal neurons and tissue, but Calhex 231 or Xesto C (a selective inhibitor of IP3 receptor) partly prevented hypoxia-induced BACE1 overexpression. Hypoxia or GdCl3 increased the content of Aß42 and Aß40 in hippocampal tissue, however Calhex 231 or Xesto C prevented hypoxia-induced the overproduction of Aß42 and Aß40 partly. Based on the above data, we suggested that hypoxia increased [Ca(2+)]i by elevated CaSR expression to promote BACE1 expression, thereby resulting in the overproduction of Aß42 and Aß40.

Development of small-molecule inhibitors that target PI3Kß.

Phosphatidylinositol-3 kinase (PI3K) ß, a subtype of class I PI3Ks, has an essential role in PTEN-deficient tumors and links to thrombosis, male fertility, and Fragile X syndrome. PI3Kß-specific targeting therapy could be an efficacious treatment for diseases highly dependent on PI3Kß, while mitigating the severe toxicity of pan-PI3K inhibitors. Achieving selectivity can be accomplished through three primary strategies, namely, binding to the induced lipophilic pocket, targeting the unique amino acid residue of PI3Kß, or using atropisomerism to lock conformation. In this review, we focus on advances in the development of these ß-isoform-selective PI3K inhibitors, providing potential guidance for the further development of novel clinical candidates.

Identification of 3-aryl-5-methyl-isoxazole-4-carboxamide derivatives and analogs as novel HIF-2α agonists through docking-based virtual screening and structural modification.

Hypoxia-inducible factor-2 (HIF-2) serves as the pivotal transcription factor in cellular responses to low oxygen levels, particularly concerning the regulation of erythropoietin (EPO) production. A docking-based virtual screening on crystal structures of HIF-2α inhibitors unexpectedly identified 3-phenyl-5-methyl-isoxazole-4-carboxamide derivative v19 as a hit of HIF-2α agonist. Further structural optimizations of compound v19 led to the discovery of a series of HIF-2α agonists with novel scaffolds. The most promising compounds 12g and 14d exhibited potent HIF-2α agonistic activities in vitro with EC50 values of 2.29 µM and 1.78 µM, respectively. Molecular dynamics simulations have revealed their capacity to allosterically enhance HIF-2 dimerization, which shed light on their mechanism of action. Moreover, compound 14d demonstrated a favorable pharmacokinetic (PK) profile, boasting an impressive oral bioavailability value of 68.71 %. These findings strongly suggest that compound 14d is an auspicious lead compound for the treatment of renal anemia.

Engineered nanovesicles from activated neutrophils with enriched bactericidal proteins have molecular debridement ability and promote infectious wound healing.

Background: Bacterial infections pose a considerable threat to skin wounds, particularly in the case of challenging-to-treat diabetic wounds. Systemic antibiotics often struggle to penetrate deep wound tissues and topically applied antibiotics may lead to sensitization, necessitating the development of novel approaches for effectively treating germs in deep wound tissues. Neutrophils, the predominant immune cells in the bloodstream, rapidly release an abundance of molecules via degranulation upon activation, which possess the ability to directly eliminate pathogens. This study was designed to develop novel neutrophil cell engineered nanovesicles (NVs) with high production and explore their bactericidal properties and application in promoting infectious wound healing. Methods: Neutrophils were isolated from peripheral blood and activated in vitro via phorbol myristate acetate (PMA) stimulation. Engineered NVs were prepared by sequentially extruding activated neutrophils followed by ultracentrifugation and were compared with neutrophil-derived exosomes in terms of morphology, size distribution and protein contents. The bactericidal effect of NVs in vitro was evaluated using the spread plate technique, LIVE/DEAD backlight bacteria assay and observation of bacterial morphology. The therapeutic effects of NVs in vivo were evaluated using wound contraction area measurements, histopathological examinations, assessments of inflammatory factors and immunochemical staining. Results: Activated neutrophils stimulated with PMA in vitro promptly release a substantial amount of bactericidal proteins. NVs are similar to exosomes in terms of morphology and particle size, but they exhibit a significantly higher enrichment of bactericidal proteins. In vitro, NVs demonstrated a significant bactericidal effect, presumably mediated by the enrichment of bactericidal proteins such as lysozyme. These NVs significantly accelerated wound healing, leading to a marked reduction in bacterial load, downregulation of inflammatory factors and enhanced collagen deposition in a full-thickness infectious skin defect model. Conclusions: We developed engineered NVs derived from activated neutrophils to serve as a novel debridement method targeting bacteria in deep tissues, ultimately promoting infectious wound healing.

Advanced degradation of refractory organic compounds in electroplating wastewater by an in-situ electro-catalytic biological coupling reactor: Removal performance, microbial community and possible mechanism.

A high-efficiency treatment system for advanced degradation of refractory organic compounds such as saccharin sodium (SS) and polyethylene glycol 6000 (PEG 6000) in electroplating wastewater was proposed, which coupled ion exchange, electrocatalysis, and microbial interactions through ion exchange particle electrode (IEPE) in a reactor, named in-situ electro-catalytic biological coupling reactor (i-SECBCR). A small-scale experimental test system was established and a feasibility investigation was conducted under the condition of 1.248 L/h continuous flow. The results revealed that (1) the i-SECBCR showed higher average removal rates of SS, PEG 6000, COD and NH4+-N, i.e. 88.48 %, 41.26 %, 66.81 % and 51.61 %,which meant an increase by 5.04 %, 12.05 %, 0.46 %, and 34.50 %, respectively, compared with BAF; (2) the optimal current intensity (CI) of i-SECBCR for simultaneous removal of SS, PEG 6000, COD and NH4+-N was 0.40 mA cm-2; (3) Rhodobacter, Defluviimonas, unclassified_f__Microscillaceae, Pseudoxanthomonas, Novosphingobium, and unclassified_f__Xanthobacteraccae accounted for the main bacterial community in i-SECBCR; (4) the possible degradation mechanism was attributed mainly to the synergistic effect of ion exchange, electrocatalytic oxidation and biology. Therefore, the i-SECBCR was suitable to simultaneously advanced remove SS, PEG 6000, COD and NH4+-N in electroplating wastewater.

Performance, community structure, metabolic pathway, and mechanism in a three-dimensional electrocatalytic biofilter (3DEBF) for the degradation of multiple concentrations of clofibric acid (CA).

A three-dimensional electrocatalytic biofilter (3DEBF) was constructed to remove clofibric acid (CA). This study compared the effectiveness of 3DEBF and biological aerated filter (BAF) in the removal of refractory CA, examined the effects of influent CA concentrations (0.1, 0.3, 0.5, 0.7, and 1.0 mg/L) on microbial community, and proposed a possible 3DEBF degradation mechanism. Results indicated that the average removal efficiency of 3DEBF reached a peak (76.09%) at 0.7 mg/L, which was 14.43% higher than that of BAF. Based on the microbial community analysis, the significant enrichment of Rhodobacter, Mycobacterium, and Sphingopyxis in 3DEBF was associated with the effect of the CA concentration and the electric field. The degradation pathway indicated that xenobiotics biodegradation and metabolism, membrane transport and replication and repair related genes were upregulated in 3DEBAF. Moreover, CA degradation is based on a combination of adsorption, electrochemical oxidation, and biodegradation.

Prospective study and validation of early warning marker discovery based on integrating multi-omics analysis in severe burn patients with sepsis.

Background: Early detection, timely diagnosis and rapid response are essential for case management and precautions of burn-associated sepsis. However, studies on indicators for early warning and intervention have rarely been conducted. This study was performed to better understand the pathophysiological changes and targets for prevention of severe burn injuries. Methods: We conducted a multi-center, prospective multi-omics study, including genomics, microRNAomics, proteomics and single-cell transcriptomics, in 60 patients with severe burn injuries. A mouse model of severe burn injuries was also constructed to verify the early warning ability and therapeutic effects of potential markers. Results: Through genomic analysis, we identified seven important susceptibility genes (DNAH11, LAMA2, ABCA2, ZFAND4, CEP290, MUC20 and ENTPD1) in patients with severe burn injuries complicated with sepsis. Through plasma miRNAomics studies, we identified four miRNAs (hsa-miR-16-5p, hsa-miR-185-5p, hsa-miR-451a and hsa-miR-423-5p) that may serve as early warning markers of burn-associated sepsis. A proteomic study indicated the changes in abundance of major proteins at different time points after severe burn injury and revealed the candidate early warning markers S100A8 and SERPINA10. In addition, the proteomic analysis indicated that neutrophils play an important role in the pathogenesis of severe burn injuries, as also supported by findings from single-cell transcriptome sequencing of neutrophils. Through further studies on severely burned mice, we determined that S100A8 is also a potential early therapeutic target for severe burn injuries, beyond being an early warning indicator. Conclusions: Our multi-omics study identified seven susceptibility genes, four miRNAs and two proteins as early warning markers for severe burn-associated sepsis. In severe burn-associated sepsis, the protein S100A8 has both warning and therapeutic effects.

The anode is more beneficial to the advanced treatment of wastewater containing antibiotics by three-dimensional electro-biofilm reactor: Degradation, mechanism and optimization.

The three-dimensional electrode biological aerated filter (3DE-BAF) has the potential to overcome inherent limitations of conventional electrochemical and biofilm methods. Electrochemical means could enhance the performance and sustainability of biofilm technologies and stimulate the spread of new applications in (waste) water treatment. This paper describes the construction and performance of 3DE-BAF in the treatment of simulated wastewater represented by tetracycline (TC). This is followed by a discussion of electrode performance, the electron transport mechanism and the electrode's effect on the biological community of 3D-EBAF. Given the gap between experimental studies and practical applications, the enlarged anode 3DE-BAF named 3DEAE-BAF reactor was applied with good results to duck farm wastewater. This study could provide guidance as to developing new methods to construct a highly stable 3DE-BAF. The paper concludes that improved 3DE-BAF technology is promising for advanced treatment of livestock wastewater containing antibiotics.

Efficient degradation of naproxen in a three dimensional biofilm electrode magnetism reactor (3DBEMR): Removal performance and microbial community.

A three-dimensional biofilm electrode magnetism reactor (3DBEMR) was constructed to removal naproxen (NPX). This study evaluated 3DBEMR performance in removal of refractory NPX, while also discussing the effect of the electro-magnetic superposition on microbial community by high throughput sequencing. Results indicated that 3DBEMR's average removal rate for NPX stood at 88.36%, representing an increase by 75.24%, 65.03% and 12.36%, respectively, compared to 3DBR (Three-Dimensional Biofilm Reactor), 3DBMR (Three-Dimensional Biofilm Magnetism Reactor) and 3DBER (Three-Dimensional Biofilm Electrode Reactor). This was attributed to the influence of electro-magnetic adsorption, electro-oxidaton/catalysis, and electro-magnetic biodegradation. Another major contributing factor to NPX removal was the presence in 3DBEMR of high-abundance genera such as Rhodobacter, Porphyrobacter, Methyloversatilis, Sphingopyxis,Bosea, Singulisphaera, Sphingomonas. Therefore, the 3DBEMR was successfully demonstrated to be a flexible and effective technique in NPX degradation, which would help to better understand the effect of superposition of electric and magnetic fields on microbial community.

Nitrogen removal performance and rapid start-up of anammox process in an electrolytic sequencing batch reactor (ESBR).

In this study, the electrolytic sequencing batch reactor (ESBR) with different current densities was constructed to investigate the nitrogen removal performance and rapid start-up of anaerobic ammonia oxidation (anammox) process. The changes of total nitrogen removal rate (TNRR), specific anammox activity (SAA) and nitrogen concentration under different current densities were analyzed, and then the effect of the optimal current density on the start-up of anammox in ESBR was explored. The results showed that ammonium nitrogen removal efficiency (92.7%), nitrite nitrogen removal efficiency (15.5%) and total nitrogen removal efficiency (28.1%) were obtained with the TNRR and SAA were 0.0118 g N L-1 d-1 and 0.0050 g N (g Vss d)-1, respectively under the optimal conditions (i.e., current density = 0.10 mA cm-2, temperature = 36 °C and pH = 7.6). In addition, the stoichiometric ratio indicated that anammox was initiated successfully for 91 days in ESBR with the current density of 0.10 mA cm-2, which was shortened by 10 days compared with the conventional SBR without current density. These results suggest that an array of rapid start-up processes of anammox can be developed through applying current density to stimulate the activity of anammox bacteria (AnAOB).

Primitive genotypic characteristics in umbilical cord neutrophils identified by single-cell transcriptome profiling and functional prediction.

The function and heterogeneity of neutrophils in neonatal umbilical cord blood (UCB) have not been characterized. In this study, we analyzed the neutrophils in UCB and healthy adults using single-cell RNA sequencing analysis for the first time. We found that neutrophils divided into six subpopulations (G2, G3, G4, G5a, G5b, and G5c) with different marker genes and different functions under homeostasis. Compared with healthy adults, neutrophils of UCB were more naïve and have more obvious degranulation and activation functions. Moreover, we found significant differences in the amount and function of G5b cells between healthy adults and UCB. The amount of G5b group in UCB was lower, but it has more degranulation, secretion and activation functions. In addition, we noted a new subset of G5c labeled by CD52, which almost did not exist in UCB. Besides, its differential genes were enriched in terms such as protein synthesis and mRNA transcription. Furthermore, uncharacteristic transcription factors ZNF-276, ZNF-319 and ZNF-354A were identified in our study. In summary, we first examined the heterogeneity and functional diversity of neutrophils in UCB, and these data provided new insights into the mechanism of neutrophil-mediated diseases of neonates and the wider use of neutrophils in UCB.

A functionalized collagen-I scaffold delivers microRNA 21-loaded exosomes for spinal cord injury repair.

MicroRNA (miRNA)-based therapies have shown great potential in the repair of spinal cord injury (SCI). MicroRNA 21 (miR21) has been proven to have an essential protective effect on SCI. However, there are some challenges for miRNAs application due to their easy degradation and ineffective cell penetration. As natural vesicles, exosomes were considered ideal carriers for miRNAs delivery for their advantages of low immunogenicity, inherent stability and tissue/cell penetration. However, poor targeting and the low capacity of specific miRNAs impede their practical applications. This study aims to develop a type of genetically engineered miR21-loaded exosomes that can be entrapped in collagen-I (Col-I) scaffold to repair SCI. The collagen-binding domain (CBD)-fused lysosome-associated membrane glycoprotein 2b (Lamp2b) protein (CBD-LP) and miR21 were overexpressed in host HEK293T (293T) cells that were used to produce engineered miR21-loaded exosomes. The CBD peptide fused in Lamp2b on the exosome surface can stably tether exosomes to Col-I scaffold, facilitate the retention of miR21-loaded exosomes in lesion sites, promote the sustained release of miR21 to cells. Finally, a functionalized Col-I scaffold biomaterial enriched with miR21-loaded exosomes was developed and it could benefit the repair of SCI. STATEMENT OF SIGNIFICANCE: MiRNA-based therapeutics have promising potential in spinal cord injury (SCI) repair. However, easy degradation and ineffective cell penetration impede miRNAs application. Exosomes are natural vehicles for miRNAs delivery but face the challenge of diffusion in vivo. Here, the collagen-binding domain (CBD)-fused Lamp2b and miR21 were overexpressed in HEK293T cells to produce miR21-loaded and CBD-modified exosomes (CBD-LP-miR21-EXOs). The CBD modified on the exosome surface can stably tether exosomes to collagen-I scaffold to form functionalized CBD-LP-miR21-EXO-Col scaffold that can facilitate the retention of miR21-loaded exosomes, promote the sustained release of miR21 to cells and finally benefit SCI repair. Furthermore, this type of functionalized collagen-I materials can be widely applied for other tissue injury repairs by enriching the CBD-LP-EXOs loaded with appropriate miRNAs.

Electrocatalysis degradation of tetracycline in a three-dimensional aeration electrocatalysis reactor (3D-AER) with a flotation-tailings particle electrode (FPE): Physicochemical properties, influencing factors and the degradation mechanism.

Novel particle electrodes, i.e. flotation tailings particle electrode (FPE), were prepared using flotation tailings, garden soil, and soluble starch with a mass ratio of 16:3:1, and then used in tetracycline wastewater treatment. The physicochemical properties of FPE were systematically characterized using SEM, XRD, FT-IR and XRF. Tetracycline adsorption and its adsorption mechanism onto FPE was explored for the first time. Parameters affecting FPE's degradation efficiency and energy consumption such as current density, electrolysis time, initial concentration, initial pH and aeration rate were examined. The electrocatalytic degradation of tetracycline shows that the degradation of tetracycline meets the pseudo-first-order kinetics. Moreover, the numbers of •OH produced on the surfaces of the cathode, anode and particle electrode were compared. Results showed that the adsorption-saturated FPE can be regenerated by electrochemical action to induce further absorption and form in-situ electrocatalysis. In order to find out the transformation products in water and degradation pathways of Tetracycline, UHPLC method was used to obtain the degradation pathways for Tetracycline. So, this work could provide a fabrication of high-efficiency and low-cost electrocatalytic for removal of pharmaceuticals pollutants from waste water as well as deeper insight into electrocatalytic mechanism, transformation products, and degradation pathways of Tetracycline in water.

The Extracellular Matrix Enriched With Exosomes for the Treatment on Pulmonary Fibrosis in Mice.

Pulmonary fibrosis (PF) is a severe respiratory disease caused by lung microenvironment changes. TGF-ß/Smad3 signaling pathway plays a critical role in the fibrotic process. MicroRNA-29 (miR-29) has proved to alleviate the occurrence of PF by downregulating TGF-ß/Smad3 signaling pathway. The miRNA application encounters obstacles due to its low stability in body and no targeting to lesions. Exosomes can be used for therapeutic delivery of miRNA due to their favorable delivery properties. However, low efficiency of separation and production impedes the therapeutic application of exosomes. In this study, we developed a liquid natural extracellular matrix (ECM) enriched with miR-29-loaded exosomes for PF treatment. The collagen-binding domain (CBD)-fused Lamp2b (CBD-Lamp2b) and miR-29 were overexpressed in human foreskin fibroblast (HFF) host cells for the entrapment of miR-29-loaded exosomes in ECM of the cells. The repeated freeze-thaw method was performed to prepare the liquid ECM enriched with exosomes without destroying the exosomal membrane. In summary, this study developed a novel functional ECM biomaterial for therapy of PF, and also provided a promising gene therapy platform for different diseases by treatment with liquid ECM that is, enriched with exosomes loaded with different functional miRNAs.

Combined effects of electrical current and nonsteroidal antiinflammatory drugs (NSAIDs) on microbial community in a three-dimensional electrode biological aerated filter (3DE-BAF).

Three-dimensional electrode biological aerated filter (3DE-BAF) with particulate bioelectrode from lithium slag was used to simultaneously remove diclofenac and clofibric acid from the synthetic domestic sewage, and the combined effects of electrical current and nonsteroidal antiinflammatory drugs (NSAIDs) on microbial community was analyzed. The results indicated that (1) the average diclofenac and clofibric acid removal efficiency in the 3DE-BAF firstly increased, attained the peak of 79.40 ± 6.74% and 69.50 ± 6.26% at 0.35 A, and then decreased to 71.82 ± 4.90% and 55.92 ± 5.17% at 0.40 A, respectively; (2) the concentration of the diclofenac and clofibric acid in 3DE-BAF gradually decreased with the increase of reactor height; (3) the current intensity and space position affected the microbial structure at the different level; (4) at the optimum current intensity, Thiothrix, Flavobacteriaceae, Halothiobacillaceae, Hydrogenophaga, and Comamonadaceae accounted for the main bacterial community for removal diclofenac and clofibric acid in the 3DE-BAF.

Enhancement on the removal of Rhodamine B (RhB) by means of the Enlarged Anode Electric Biological (EAEB) reactor.

The Enlarged Anode Electric Biological (EAEB) Reactor was proposed, as an alternative to the common BAF. The goal of this research was to develop a new process for simultaneously removing, NH4+-N and Rhodamine B (RhB) from dyeing wastewater. The performance of EAEB was evaluated based on COD, NH4+-N and RhB removal efficiency in the effluent. The study found that the removal rate of RhB, which is a characteristic of the inoculation and start period, reaches 80% in EAEB and 30% in common BAF. A current intensity of 0.5 A, HRT of 3.5 h, and electrode area of 0.13 m2 were identified as operating parameters that could guarantee excellent RhB removal efficiency. It is worth noting that the removal of RhB in the two reactors was mainly concentrated in the 80 cm-140 cm area (measuring upwards at the top of the support layer). The removal rate of EAEB in this area was 97.7%, and the common BAF was 84.3%. Besides, in each segment of EAEB, the removal effect of RhB was better than in common BAF. This study elucidated the synergistic effects of electricity and biofilm on contaminant removal and identified important roles of improvements to the anode electro-biodegradation process. As compared to conventional technologies, the proposed process provides a highly efficient new alternative to dyeing wastewater treatment technology.

Single cell derived spheres of umbilical cord mesenchymal stem cells enhance cell stemness properties, survival ability and therapeutic potential on liver failure.

Umbilical cord mesenchymal stem cells (UCMSCs) have shown great potentials in regenerative medicine for their extensive sources, multilineage differentiation potential, low immunogenicity and self-renewal ability. However, the clinical application of UCMSCs still confronts many challenges including the requirement of large quantity of cells, low survival ability in vivo and the loss of main original characteristics due to two-dimensional (2D) culture. The traditional three-dimensional (3D)-spheroid culture can mimic in vivo conditions, but still has limitations in clinical application due to large size of spheroid against direct injection and inner cell death. Based on self-renewal tenet, we produced single cell derived sphere (SCDS) of UCMSCs through combining single cell pattern on chip with 3D culture. Compared with the 2D and traditional 3D culture, SCDS culture has many advantages to meet clinical requirements, including small size, higher abilities of survival and migration, and stronger hypoxia resistance and stemness maintenance. Furthermore, SCDS culture promotes angiogenesis in UCMSCs-xenografts and displays greater therapeutic potential on acute liver failure (ALF) in vivo. Our results suggest that SCDS culture may serve as a simple and effective strategy for UCMSCs optimization to meet clinical demand.

Removal of diclofenac by three-dimensional electro-Fenton-persulfate (3D electro-Fenton-PS).

Diclofenac (DIC) is a new type of contaminant that has been widely detected in the water environment, posing threats to the ecological environment and human health. However, the conventional wastewater treatment process has a very limited ability to reduce DIC. In this research, persulfate is added to electro-Fenton with the three-dimensional particle electrode (TDE) process whose particle electrodes were formed from manganese slag with loaded active substance (Cu: Fe = 1:1) to construct a three-dimensional electro-Fenton-persulfate (3D electro-Fenton-PS) process to investigate the removal rate of DIC under the optimum working conditions. The effects of different persulfate addition, activator addition and different activators on the removal rate of DIC were researched, respectively. The removal rate of DIC reached 96.3% when the persulfate and the Fe0 addition were 1.50 mM and 3.00 mM, respectively. The results showed that and OH existed simultaneously in the reaction system, and the removal of DIC was the result of the two free radicals. Moreover, degradation pathways and mechanism of DIC were also discussed. The study may provide a new theoretical basis and technical support for the treatment of DIC in municipal wastewater.