Immobilization of cross-linked enzymes aggregates on hierarchical covalent organic frameworks: Highly stable chemoenzymatic nanoreactor for asymmetric synthesis of optically active halohydrins.

Organometallic catalyst is extensively applied for the non-enzymatic regeneration of nicotinamide adenine dinucleotide (phosphate) cofactors, but suffering from the mutual inactivation with the enzymes in one pot. The spatially separated immobilization of organometallic catalyst and enzymes on suitable carriers not only can reduce their mutual inhabitation but also can enhance their reusability. Here in this work, we present a hierarchical porous COFs (HP-TpBpy) that incorporated with [(Cp*RhCl2]2 to generate the metalized COF, Rh-HP-TpBpy. The obtained Rh-HP-TpBpy exhibited superior performance in nicotinamide adenine dinucleotide (NADH) or nicotinamide adenine dinucleotide phosphate (NADPH) regeneration using formate as the hydride donor, significantly outperforming the natural formate dehydrogenases in cofactor preference toward NADP+. Subsequently, the Lactobacillus fermentum short-chain dehydrogenase/reductase 1 (LfSDR1) was then cross-linked into enzyme aggregates (CLEA) and immobilized on hierarchical Rh-HP-TpBpy, achieving the integrated chemoenzymatic catalyst, LfSDR1@Rh-HP-TpBpy, which can catalyze the chemoenzymatic reduction of halogenated aryl ketones and give the corresponding optically active halohydrins with high conversion and enantiomeric excess (ee) value up to 99 %. The LfSDR1@Rh-HP-TpBpy also exhibits largely enhanced stability compared with the free LfSDR1 and the CLEAs-LfSDR1, enabling its excellent reusability.

Microfluidic Synthesis of Multifunctional Micro-/Nanomaterials from Process Intensification: Structural Engineering to High Electrochemical Energy Storage.

Multifunctional micro-/nanomaterials featuring functional superiority and high value-added physicochemical nature have received immense attention in electrochemical energy storage. Microfluidic synthesis has become an emergent technology for massively producing multifunctional micro-/nanomaterials with tunable microstructure and morphology due to its rapid mass/heat transfer and precise fluid controllability. In this review, the latest progresses and achievements in microfluidic-synthesized multifunctional micro-/nanomaterials are summarized via reaction process intensification, multifunctional micro-/nanostructural engineering and electrochemical energy storage applications. The reaction process intensification mechanisms of various micro-/nanomaterials, including quantum dots (QDs), metal materials, conducting polymers, metallic oxides, polyanionic compounds, metal-organic frameworks (MOFs) and two-dimensional (2D) materials, are discussed. Especially, the multifunctional structural engineering principles of as-fabricated micro-/nanomaterials, such as vertically aligned structure, heterostructure, core-shell structure, and tunable microsphere, are introduced. Subsequently, the electrochemical energy storage application of as-prepared multifunctional micro-/nanomaterials is clarified in supercapacitors, lithium-ion batteries, sodium-ion batteries, all-vanadium redox flow batteries, and dielectric capacitors. Finally, the current problems and future forecasts are illustrated.

Modularization of Immobilized Multienzyme Cascades for Continuous-Flow Enantioselective C-H Amination.

Multienzyme cascades (MECs) have gained much attention in synthetic chemistry but remain far from being a reliable synthetic tool. Here we report a four-enzyme cascade comprising a cofactor-independent and a cofactor self-sustaining bienzymatic modules for the enantioselective benzylic C-H amination of arylalkanes, a challenging transformation from bulk chemicals to high value-added chiral amines. The two modules were subsequently optimized by enzyme co-immobilization with microenvironmental tuning, and finally integrated in a gas-liquid segmented flow system, resulting in simultaneous improvements in enzyme performance, mass transfer, system compatibility, and productivity. The flow system enabled continuous C-H amination of arylalkanes (up to 100 mM) utilizing the sole cofactor NADH (0.5 mM) in >90 % conversion, achieving a high space-time yield (STY) of 3.6 g ⋅ L-1 ⋅ h-1, which is a 90-fold increase over the highest value previously reported.

Integrating Au Catalysis and Engineered Amine Dehydrogenase for the Chemoenzymatic Synthesis of Chiral Aliphatic Amines.

Direct synthesis of aliphatic amines from alkynes is highly desirable due to its atom economy and high stereoselectivity but still challenging, especially for the long-chain members. Here, a combination of Au-catalyzed alkyne hydration and amine dehydrogenase-catalyzed (AmDH) reductive amination was constructed, enabling sequential conversion of alkynes into chiral amines in aqueous solutions, particularly for the synthesis of long-chain aliphatic amines on a large scale. The production of chiral aliphatic amines with more than 6 carbons reached 36-60 g/L. A suitable biocatalyst [PtAmDH (A113G/T134G/V294A)], obtained by data mining and active site engineering, enabled the transformation of previously inactive long-chain ketones at high concentrations. Computational analysis revealed that the broader substrate scope and tolerance with the high substrate concentrations resulted from the additive effects of mutations introduced to the three gatekeeper residues 113, 134, and 294.

Construction of Multi-Enzyme Integrated Catalysts for Deracemization of Cyclic Chiral Amines.

Multi-enzyme cascade catalysis has become an important technique for chemical reactions used in manufacturing and scientific study. In this research, we designed a four-enzyme integrated catalyst and used it to catalyse the deracemization reaction of cyclic chiral amines, where monoamine oxidase (MAO) catalyses the enantioselective oxidation of 1-methyl-1,2,3,4-tetrahydroisoquinoline (MTQ), imine reductase (IRED) catalyses the stereo selective reduction of 1-methyl-3,4-dihydroisoquinoline (MDQ), formate dehydrogenase (FDH) is used for the cyclic regeneration of cofactors, and catalase (CAT) is used for decomposition of oxidative reactions. The four enzymes were immobilized via polydopamine (PDA)-encapsulated dendritic organosilica nanoparticles (DONs) as carriers, resulting in the amphiphilic core-shell catalysts. The hydrophilic PDA shell ensures the dispersion of the catalyst in water, and the hydrophobic DON core creates a microenvironment with the spatial confinement effect of the organic substrate and the preconcentration effect to enhance the stability of the enzymes and the catalytic efficiency. The core-shell structure improves the stability and reusability of the catalyst and rationally arranges the position of different enzymes according to the reaction sequence to improve the cascade catalytic performance and cofactor recovery efficiency.

Molecular Engineering and Morphology Control of Covalent Organic Frameworks for Enhancing Activity of Metal-Enzyme Cascade Catalysis.

Metal-enzyme integrated catalysts (MEICs) that combine metal and enzyme offer great potential for sustainable chemoenzymatic cascade catalysis. However, rational design and construction of optimal microenvironments and accessible active sites for metal and enzyme in individual nanostructures are necessary but still challenging. Herein, Pd nanoparticles (NPs) and Candida antarctica lipase B (CALB) are co-immobilized into the pores and surfaces of covalent organic frameworks (COFs) with tunable functional groups, affording Pd/COF-X/CALB (X = ONa, OH, OMe) MEICs. This strategy can regulate the microenvironment around Pd NPs and CALB, and their interactions with substrates. As a result, the activity of the COF-based MEICs in catalyzing dynamic kinetic resolution of primary amines is enhanced and followed COF-OMe > COF-OH > COF-ONa. The experimental and simulation results demonstrated that functional groups of COFs modulated the conformation of CALB, the electronic states of Pd NPs, and the affinity of the integrated catalysts to the substrate, which contributed to the improvement of the catalytic activity of MEICs. Further, the MEICs are prepared using COF with hollow structure as support material, which increased accessible active sites and mass transfer efficiency, thus improving catalytic performance. This work provides a blueprint for rational design and preparation of highly active MEICs.

Zirconium-containing nanoscale coordination polymers for positron emission tomography and fluorescence-guided cargo delivery to triple-negative breast tumors.

Nanoscale coordination polymer (NCP) is a class of hybrid materials formed by self-assembly of metal ions and organic ligands through coordination. The applications of NCP in biomedicine are quite extensive due to the diversity choice of metal ions and organic ligands. Here we designed Zr-P1 NCP based on Zr4+ selected as metal ion nodes and tetrakis(4-carboxyphenyl) ethylene as bridging ligands. Zr-P1 NCP was modified with functionalized pyrene derived polyethylene glycol (Py-PAA-PEG-Mal) on the surface and further conjugated with cRGD for active targeting of integrin αvß3 overexpressed in triple-negative breast cancer. Doxorubicin was loaded on Zr-P1 NCP with encapsulation efficiency up to 22 % for the treatment of triple negative breast cancer. 89Zr-P1 NCP can be used for in vivo tumor imaging due to the fluorescence properties resulting from the enhanced aggregation-induced Emission (AIE) behavior of P1 ligands and its positron emission tomography (PET) capability. Cellular evaluation indicated that the functionalized Zr-P1@PEG-RGD presented a good function for tumor cell targeting imaging and doxorubicin could be targeted to triple negative breast cancer when it was loaded onto Zr-P1@PEG-RGD, which corroborated with the in vivo results. In summary, 89Zr-P1@PEG-RGD can serve as a biocompatible nanoplatform for fluorescence and PET image-guided cargo delivery. STATEMENT OF SIGNIFICANCE: Nanoscale coordination polymer (NCP) is a class of hybrid materials formed by self-assembly of metal ions and organic ligands through coordination. The diversity of available metals and ligand structures upon NCP synthesis plays an advantage in establishing multimodal imaging platforms. Here we designed 89Zr-P1@PEG-RGD NCP based on Zr4+ selected as metal ion nodes and tetrakis(4-carboxyphenyl) ethylene as bridging ligands. 89Zr-P1@PEG-RGD nanomaterials have positron emission tomography (PET) capability due to the incorporation of zirconium-89, which can be used for in vivo tumor imaging with high sensitivity. The chemotherapeutic drug DOX was loaded on Zr-P1 NCP for the treatment of triple-negative breast cancer, and dual modality imaging can provide visual guidance for drug delivery.

Synergistic Activation of Electric Furnace Ferronickel Slag by Mechanical Grinding and Chemical Activators to Prepare Cementitious Composites.

The use of electric furnace ferronickel slag (FNS) as a supplementary cementitious material is the current focus of research. This study investigates the effect of mechanical grinding and chemical additives on the activity excition of FNS, as well as the associated synergistic mechanisms. This study shows that the addition of triethanolamine (TEA) increases the fine-grained content in FNS powder, which facilitates the depolymerization of FNS and the early hydration of aluminum tricalcium. Furthermore, the addition of Ca(OH)2 raises the alkalinity of the cementitious system, which promotes the availability of Ca2+ ions and accelerates the hydration process, resulting in the generation of additional hydration products. The enhancement of late hydration of C3S by TEA and its combination with the secondary hydration of Ca2+ at high alkalinity are the pivotal factors to improve the strength of cementitious composite. A mixture of FNS and 0.03% TEA is subjected to grinding for 90 min, using the obtained micropowder which replaces 20% of the cement, and subsequently, after being excited with 3% Ca(OH)2, the FNS micropowder reaches the quality standards of S95 slag powder. It is worth remarking that the micropowder prepared by mixing FNS with 3% Ca(OH)2 and 0.03% TEA and grinding it for 81 min also meets the S95 standard for slag powder. The larger dosage of FNS in cement is supported by the observed synergy between TEA and Ca(OH)2. This research will provide valuable insights for the expanded application of FNS in construction materials.

Resin-Immobilized Palladium Acetate and Alcohol Dehydrogenase for Chemoenzymatic Enantioselective Synthesis of Chiral Diarylmethanols.

The enantioselective synthesis of chiral diarylmethanols is highly desirable in synthetic chemistry and the pharmaceutical industry, but it remains challenging, especially in terms of green and sustainable production. Herein, a resin-immobilized palladium acetate catalyst was fabricated with high activity, stability, and reusability in Suzuki cross-coupling reaction of acyl halides with boronic acids, and the coimmobilization of alcohol dehydrogenase and glucose dehydrogenase on resin supports was also conducted for asymmetric bioreduction of diaryl ketones. Experimental results revealed that the physicochemical properties of the resins and the immobilization modes played important roles in affecting their catalytic performances. These two catalysts enabled the construction of a chemoenzymatic cascade for the enantioselective synthesis of a series of chiral diarylmethanols in high yields (83-90%) and enantioselectivities (87-98% ee). In addition, the asymmetric synthesis of the antihistaminic and anticholinergic drugs (S)-neobenodine and (S)-carbinoxamine was also achieved from the chiral diarylmethanol precursors, demonstrating the synthetic utility of the chemoenzymatic cascade.

Heteroatom Structural Engineering of Conjugated Porous Polymers Enhances Photocatalytic Nicotinamide Cofactor Regeneration.

Photocatalysis is an eco-friendly method to regenerate nicotinamide (NADH) cofactors, which is essential for biotransformation over oxidoreductases. Organic polymers exhibit high stability, biocompatibility and functional designability as photocatalysts, but still suffering from rapid charge recombination. Herewith the heteroatom structural engineering of donor-π-acceptor (D-π-A) conjugated porous polymers were conducted to promote charge transfer and photocatalytic NADH regeneration. The electron delocalization of polymer photocatalysts can be readily tuned by changing the electron density of the donor unit, leading to faster charge separation and better photocatalytic performance. The optimum sulfur-doped polymer exhibits the highest NADH regeneration yield of 47.4 % in 30 min and 94.1 % in 4 h, which can drive the biocatalytic C=C bond reduction of 2-cyclohexen-1-one by ene-reductase, giving the corresponding cyclohexanone yield of 96.7 % in 10 h. Moreover, the oxygen-doped polymer, from biomass derived 2,5-diformylfuran, exhibits comparable photocatalytic activity to the sulfur-doped CPP, suggesting the potential of furan as alternative donor unit to thiophene.

Activation Mechanism of Ammonium Fluoride in Facile Synthesis of Hydrated Silica Derived from Ferronickel Slag-Leaching Residue.

A novel process for the synthesis of hydrated silica derived from ferronickel slag (FNS)-leaching residue was proposed in this study. The products of the purification of hydrated silica with 99.68% grade and 95.11% recovery can be obtained through ammonium fluoride (NH4F) roasting, followed by the process of water leaching, ammonia precipitating, and acid cleaning under the optimized conditions. The effects of NH4F mass ratio, roasting temperature, and roasting time on the water-leaching efficiency were investigated in detail. The thermodynamic and X-ray diffraction analyses indicated that the amorphous silica in FNS-leaching residue was converted to water-soluble fluoride salts ((NH4)2SiF6) during the roasting process, which are also supported by the scanning electron microscopy and thermogravimetry analyses. The Si-O bonds in amorphous silica could be effectively broken through the ammonium fluoride activation during a low-temperature roasting process. This work provides a meaningful reference for further studies on the facile synthesis of hydrated silica with similar mineral compositions.

Asymmetric α-benzylation of cyclic ketones enabled by concurrent chemical aldol condensation and biocatalytic reduction.

Chemoenzymatic cascade catalysis has emerged as a revolutionary tool for streamlining traditional retrosynthetic disconnections, creating new possibilities for the asymmetric synthesis of valuable chiral compounds. Here we construct a one-pot concurrent chemoenzymatic cascade by integrating organobismuth-catalyzed aldol condensation with ene-reductase (ER)-catalyzed enantioselective reduction, enabling the formal asymmetric α-benzylation of cyclic ketones. To achieve this, we develop a pair of enantiocomplementary ERs capable of reducing α-arylidene cyclic ketones, lactams, and lactones. Our engineered mutants exhibit significantly higher activity, up to 37-fold, and broader substrate specificity compared to the parent enzyme. The key to success is due to the well-tuned hydride attack distance/angle and, more importantly, to the synergistic proton-delivery triade of Tyr28-Tyr69-Tyr169. Molecular docking and density functional theory (DFT) studies provide important insights into the bioreduction mechanisms. Furthermore, we demonstrate the synthetic utility of the best mutants in the asymmetric synthesis of several key chiral synthons.

Mechanism of adipose tissue-derived stromal cell-extracellular vesicles in treating oral submucous fibrosis by blocking the TGF-ß1/Smad3 pathway via the miR-760-3p/IGF1R axis

Oral submucous fibrosis (OSF) is a prevalent chronic condition, and understanding its pathogenesis is crucial for developing effective therapeutic strategies. This study explores the potential of adipose tissue-derived stromal cell-extracellular vesicles (ADSC-EVs) in mitigating OSF and investigates the underlying molecular mechanisms. OSF was induced in mice by arecoline feeding. Adipose tissue-derived stromal cells (ADSCs), fibrotic buccal mucosal fibroblasts (fBMFs) isolated from OSF mice, and ADSC-EVs were comprehensively characterized. The treatment effects of extracellular vesicles (EVs) and pcDNA3.1-IGF1R on fBMF proliferation, migration, and invasion were assessed using Cell Counting Kit-8 (CCK-8) assay, transwell assay, and flow cytometry assay. The expression levels of alpha smooth muscle actin (α-SMA), collagen I, collagen III, and insulin-like growth factor 1 receptor (IGF1R) were evaluated by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot. The interaction between miR-760-3p and IGF1R was investigated. In fBMFs and OSF mice treated with a miR-760-3p inhibitor and/or EVs, the expression patterns of miR-760-3p, IGF1R, and proteins related to the TGF-ß1/Smad3 pathway were determined. ADSC-EVs demonstrated the ability to upregulate miR-760-3p, impede cell proliferation, migration, and invasion, and reduce α-SMA, collagen I, and collagen III levels in fBMFs. The expression of miR-760-3p was diminished in ADSC-EVs treated with a miR-760-3p inhibitor. However, silencing miR-760-3p or overexpressing IGF1R partially counteracted the beneficial effects of ADSC-EVs on fBMF fibrosis. miR-760-3p directly targets IGF1R. Significantly, ADSC-EVs exert their suppressive effects on the TGF-ß1/Smad3 pathway through the miR-760-3p/IGF1R axis. In summary, ADSC-EVs, by transferring miR-760-3p and inhibiting IGF1R expression, effectively block the TGF-ß1/Smad3 pathway, thereby alleviating fibrosis in fBMFs and preventing the progression of OSF.

How environmental decentralization affects the synergy of pollution and carbon reduction: Evidence based on pig breeding in China.

Reducing pollution and carbon is essential to achieve China's goal of "carbon peaking and carbon neutrality"; however, the collaborative paths of pollution and carbon reduction remain vague and worth exploring. This paper analyses panel data from 30 provinces in China from 2002 to 2017 to determine the impact of environmental decentralization on the synergy of pollution and carbon reduction in pig farming. The result shows that environmental decentralization has a significant 'carbon reduction effect' and 'pollution reduction effect' on pig farming; it is also conducive to promoting the synergistic effect of reducing pollution and carbon emissions through supporting environmental facilities and industrial organisations. Various types of environmental decentralization have significant differences in the synergy of pollution and carbon reduction. The scale of pig breeding plays a positive regulatory role in the impact of environmental decentralization on the synergy of pollution and carbon reduction while showing regional heterogeneity. This research is crucial for advancing the green transformation of pig breeding.

Engineered baicalein-decorated zinc phosphates for synergistic alleviation of inflammatory bowel disease by repairing the mucosal barrier and relieving oxidative stress.

Orally administered baicalein-decorated zinc phosphates (ZnBM) were engineered for mucosal barrier improvement and intestinal inflammation relief. ZnBM with a size of 1.78 µm comprised 5.58 wt% baicalein and 13.17 wt% zinc. The incorporation of baicalein endowed ZnBM with excellent radical scavenging activities. ZnBM exhibited good stability with negligible zinc release in PBS solution for 2 days, and 32.82% of the zinc could reach the gut. In addition, ZnBM polarized macrophages into the anti-inflammatory M2 type and effectively scavenged intracellular reactive oxygen species (ROS) of lipopolysaccharide (LPS)-treated RAW264.7. Meanwhile, ZnBM effectively scavenged intracellular ROS of phorbol 12-myristate 13-acetate (PMA)-induced Caco-2 cells and exerted a reparative effect on the LPS-damaged Caco-2 monolayer, causing an obvious improvement of the barrier function. Reduced systemic exposure to FITC-dextran was observed to illustrate barrier restoration by ZnBM, which was achieved through upregulation of tight junction protein expression. Notably, the commonly used clinical drug 5-aminosalicylic acid is toxic to the liver and kidneys, and commercial ZnO caused the death of mice during treatment. Apparently, the therapeutic effect of ZnBM was significantly better than that of baicalein alone in chronic colitis. Overall, ZnBM exhibited outstanding therapeutic efficacy and is expected to treat colitis due to its effectiveness, biosecurity, facile preparation, and easy storage.

Identification of Fangchinoline as a novel autophagy inhibitor with an adjuvant of chemotherapy against lung cancer.

Autophagy is a fundamental recycling pathway that enhances cellular resilience, promoting survival. However, this survival mechanism can impede anti-cancer treatment strategies designed to induce cell death. In this study, we identified a novel autophagy inhibitor, Fangchinoline (Fan) isolated from the traditional Chinese medicine Stephania tetrandra. We speculated that when Fan blocks autophagy, cancer cells lose substantial self-preservation abilities during treatment. Firstly, we examined in detail the mechanism through which Fan inhibits autophagy. Specifically, Fan induced a significant increase in autophagosomes, as indicated by GFP-LC3 labeling, confirmed by the up-regulation of LC3-II. The autophagy receptor protein p62 was also up-regulated, suggesting a potential inhibition of autophagy flux. We further ruled out the possibility of fusion barriers between lysosomes and autophagosomes, as confirmed by their co-localization in double fluorescence staining. However, the lysosomal acid environment might be compromised, as suggested by the diminished fluorescence of acidity-sensitive dyes in the lysosomes and the corresponding decrease in mature forms of lysosomal cathepsin. To test the anti-cancer potential of Fan, we combined it with Cisplatin (Cis) or Paclitaxel (PTX) for lung cancer cell treatment. This combined treatment demonstrated a synergistically enhanced killing effect. These promising anti-tumor results were also replicated in a xenografted tumor model. The significance of this research lies in the identification of Fan as a potent autophagy inhibitor and its potential to enhance the efficacy of existing anti-cancer drugs. By unraveling the mechanisms of Fan's action on autophagy and demonstrating its synergistic effect in combination therapies, our study provides valuable insights for developing novel strategies to overcome autophagy-mediated resistance in cancer treatment.

Identification of Xanthine Oxidase Inhibitors from Celery Seeds Using Affinity Ultrafiltration-Liquid Chromatography-Mass Spectrometry.

Celery seeds have been used as an effective dietary supplement to manage hyperuricemia and diminish gout recurrence. Xanthine oxidase (XOD), the critical enzyme responsible for uric acid production, represents the most promising target for anti-hyperuricemia in clinical practice. In this study, we aimed to establish a method based on affinity ultrafiltration-liquid chromatography-mass spectrometry (UF-LC-MS) to directly and rapidly identify the bioactive compounds contributing to the XOD-inhibitory effects of celery seed crude extracts. Chemical profiling of celery seed extracts was performed using UPLC-TOF/MS. The structure was elucidated by matching the multistage fragment ion data to the database and publications of high-resolution natural product mass spectrometry. Thirty-two compounds, including fourteen flavonoids and six phenylpeptides, were identified from celery seed extracts. UF-LC-MS showed that luteolin-7-O-apinosyl glucoside, luteolin-7-O-glucoside, luteolin-7-O-malonyl apinoside, luteolin-7-O-6'-malonyl glucoside, luteolin, apigenin, and chrysoeriol were potential binding compounds of XOD. A further enzyme activity assay demonstrated that celery seed extract (IC50 = 1.98 mg/mL), luteolin-7-O-apinosyl glucoside (IC50 = 3140.51 µmol/L), luteolin-7-O-glucoside (IC50 = 975.83 µmol/L), luteolin-7-O-6'-malonyl glucoside (IC50 = 2018.37 µmol/L), luteolin (IC50 = 69.23 µmol/L), apigenin (IC50 = 92.56 µmol/L), and chrysoeriol (IC50 = 40.52 µmol/L) could dose-dependently inhibit XOD activities. This study highlighted UF-LC-MS as a useful platform for screening novel XOD inhibitors and revealed the chemical basis of celery seed as an anti-gout dietary supplement.

Zinc-Rutin Particles Ameliorate DSS-Induced Acute and Chronic Colitis via Anti-inflammatory and Antioxidant Protection of the Intestinal Epithelial Barrier.

In patients suffering from inflammatory bowel diseases (IBDs), the immune system is disrupted and the intestinal barrier function is compromised. Here, six zinc-flavonoid particles were produced by one-step reaction via changing flavonoids (myricetin, quercetin, and rutin) and solvent (water and ethanol), and then their cytocompatibility and ability to scavenge H2O2, free radicals, and LPS-induced ROS were compared. Zinc-rutin particles (W-ZnRT) composed of rutin (78.92 wt %), Na12[ZnPO4]12·12H2O (6.76 wt %), and crystal water were screened out because W-ZnRT exhibited 80.8 ± 15% cell viability against RAW264.7, could rapidly scavenge 78.1 ± 1% of H2O2 and 71.6 ± 2% of DPPH within 30 min, and reduced LPS-increased intracellular ROS to normal levels. In addition, the therapeutic effects of rutin and W-ZnRT were also compared in dextran sulfate sodium (DSS)-induced acute and chronic colitis in mice. W-ZnRT was superior to rutin alone in chronic colitis (n = 9), although they were equally effective in acute colitis (n = 7). Compared to rutin, 11 oral doses of W-ZnRT (40 mg kg-1) significantly improved intestinal permeability (p = 0.0299) and colon length (p = 0.0025), reduced intestinal proinflammatory factors (IL-6, IL-1ß, and TNF-α), and upregulated tight junction proteins to maintain intestinal barrier function. Taken together, these results identified W-ZnRT as an efficient and safe therapeutic strategy for IBD.

Transfer of patient's peripheral blood mononuclear cells (PBMCs) disrupts blood-brain barrier and induces anti-NMDAR encephalitis: a study of novel humanized PBMC mouse model.

BACKGROUND: Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe autoimmune neuropsychiatric disease. Brain access of anti-NMDAR autoantibody through the blood-brain barrier (BBB) is essential for pathogenesis. Most previous animal models limit the investigation of etiologies of BBB damage in patients. METHODS: In this study, we established a novel humanized mouse model of anti-NMDAR encephalitis by intraperitoneal injection of patients' peripheral blood mononuclear cells (PBMCs) into BALB/c Rag2-/-Il2rg-/-SirpαNODFlk2-/- mice. RESULTS: We found that engraftment of patients' PBMCs not only produced potent anti-GluN1 autoantibodies, but also disrupted BBB integrity to allow brain access of autoantibodies, resulting in a hyperactive locomotor phenotype, anxiety- and depressive-like behaviors, cognitive deficits, as well as functional changes in corresponding brain regions. Transcriptome analysis suggested an exaggerated immune response and impaired neurotransmission in the mouse model and highlighted Il-1ß as a hub gene implicated in pathological changes. We further demonstrated that Il-1ß was produced by endothelial cells and disrupted BBB by repressing tight junction proteins. Treatment with Anakinra, an Il-1 receptor antagonist, ameliorated BBB damage and neuropsychiatric behaviors. CONCLUSIONS: Our study provided a novel and clinically more relevant humanized mouse model of anti-NMDAR encephalitis and revealed an intrinsic pathogenic property of the patient's lymphocytes.

Identification of Fangjihuangqi Decoction as a late-stage autophagy inhibitor with an adjuvant anti-tumor effect against non-small cell lung cancer.

BACKGROUND: Clinically, although chemotherapy is one of the most commonly used methods of treating tumors, chemotherapeutic drugs can induce autophagic flux and increase tumor cell resistance, leading to drug tolerance. Therefore, theoretically, inhibiting autophagy may improve the efficacy of chemotherapy. The discovery of autophagy regulators and their potential application as adjuvant anti-cancer drugs is of substantial importance. In this study, we clarified that Fangjihuangqi Decoction (FJHQ, traditional Chinese medicine) is an autophagy inhibitor, which can synergistically enhance the effect of cisplatin and paclitaxel on non-small cell lung cancer (NSCLC) cells. METHODS: We observed the changes of autophagy level in NSCLC cells under the effect of FJHQ, and verified the level of the autophagy marker protein and cathepsin. Apoptosis was detected after the combination of FJHQ with cisplatin or paclitaxel, and NAC (ROS scavenger) was further used to verify the activation of ROS-MAPK pathway by FJHQ. RESULTS: We observed that FJHQ induced autophagosomes in NSCLC cells and increased the levels of P62 and LC3-II protein expression in a concentration- and time-gradient-dependent manner, indicating that autophagic flux was inhibited. Co-localization experiments further showed that while FJHQ did not inhibit autophagosome and lysosome fusion, it affected the maturation of cathepsin and thus inhibited the autophagic pathway. Finally, we found that the combination of FJHQ with cisplatin or paclitaxel increased the apoptosis rate of NSCLC cells, due to increased ROS accumulation and further activation of the ROS-MAPK pathway. This synergistic effect could be reversed by NAC. CONCLUSION: Collectively, these results demonstrate that FJHQ is a novel late-stage autophagy inhibitor that can amplify the anti-tumor effect of cisplatin and paclitaxel against NSCLC cells.