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Additional file 3: Table S2. Raw reads of the top ten differentially expressed miRNAs from deep-sequencing. The raw data of the top ten differentially expressed miRNAs between MCS and MCD group from deep-sequencing are listed in Table S2. FDR: false discovery rate, FC: fold change.

Additional file 1: Figure S1. Body weight, liver weight and circulating lipid content in MCD and HFD-induced NAFLD mice. Mice were fed a MCD diet for 4 weeks or HFD for 16 weeks to induce NAFLD. (A) Body weight of NAFLD mice, n = 6 per group. (B) Representative images of liver and the liver weight of NAFLD mice, n = 6 per group. Scale bar:5 mm. (C-E) Relative plasma total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) content, n = 6 per group. Data are expressed as the mean ± SEM. Statistics: Student t-test, *P < 0.05, **P < 0.01 vs. the MCS group; &&P < 0.01 vs. ND group. Figure S2. Changes of microvascular endothelial permeability in lung, liver and spleen induced by hepatic sEVs. NAFLD or control hepatic sEVs were isolated by ultracentrifugation, identified, and administered to naive NLRP3+/+ and NLRP3-/- mice via caudal vein injection. (A-C) In vivo optical imaging system-obtained fluorescence imaging of lung, liver and spleen, n = 6 per group. The pellet derived from the ultracentrifugation of DiR alone was used as a vehicle control. (D-F) Representative images and the summarized data of Evans blue concentrations in lung, liver and spleen, n = 5-6 per group. Data are expressed as the mean ±SEM. Statistics: One-way ANOVA, *P < 0.05 vs. NLRP3+/+ mice injected with MCS hepatic sEVs; &&P < 0.01 vs. NLRP3+/+ mice injected with ND hepatic sEVs; ## P < 0.01 vs. NLRP3+/+ mice injected with MCD hepatic sEVs; $$ P < 0.01 vs. NLRP3+/+ mice injected with HFD hepatic sEVs. Figure S3. Cell viability, LDH leakage and lipid accumulation in steatotic hepatocytes. Human hepatocyte cell line HepG2, HuH7 and L02 were treated with 100 μmol/L palmitic acid (PA) or vehicle for 18 h. (A) Cell viability were measured using cell counting kit (CCK)-8, n = 6 per group. (B) Lactic dehydrogenase (LDH) leakage was estimated by measuring LDH activities in the cell culture supernatants, n = 6 per group. (C) Intracellular triglyceride (TG) contents were measured by chromometry, n = 4 per group. (D) Hepatocytes were stained with Oil Red O and the intracellular lipid accumulation was measured by chromometry, n = 4 per group. Data are expressed as the mean ±SEM. Statistics: Student t-test, Ns, no significance, **P < 0.01 vs. vehicle group. Figure S4. Cytokine contents in the cardiac tissue of NAFLD mice.NLRP3+/+ and NLRP3-/- Mice were fed a MCD diet for 4 weeks or HFD for 16 weeks to induce NAFLD. The contents of cytokines including IL-1β and HMGB1 in the cardiac tissues were measured by ELISA kits. The results were normalized to protein concentration. (A-B) Cardiac IL-1β and HMGB1 content in NLRP3+/+ and NLRP3-/- mice fed a MCS or MCD diet, n = 5-6 per group. (C-D) Cardiac IL-1β and HMGB1 content in NLRP3+/+ and NLRP3-/- Mice fed a ND or HFD, n = 5-6 per group. Data are expressed as the mean ±SEM. Statistics: One-way ANOVA, **P < 0.01 vs. NLRP3+/+ mice fed with a HFD diet. Figure S5. Cytokine contents in the cardiac tissue of hepatic. sEV-treated mice NAFLD or control hepatic sEVs were isolated by ultracentrifugation, identified, and administered to naive NLRP3+/+ and NLRP3-/- mice via caudal vein injection. The contents of cytokines including IL-1β and HMGB1 in the cardiac tissues were measured by ELISA kits. The results were normalized to protein concentration. (A-B) Cardiac IL-1β and HMGB1 content in NLRP3+/+ and NLRP3-/- mice injected with MCS-sEVs or MCD-sEVs, n = 5-6 per group. (C-D) Cardiac IL-1β and HMGB1 content in NLRP3+/+ and NLRP3-/- mice injected with ND-sEVs or HFD-sEVs, n = 5-6 per group. Data are expressed as the mean ±SEM. Statistics: One-way ANOVA, **P < 0.01 vs. NLRP3+/+ mice injected with MCS-sEVs; ##P < 0.01 vs. NLRP3+/+ mice injected with MCD-sEVs; &&P < 0.01 vs. NLRP3+/+ mice injected with ND-sEVs; $$P < 0.01 vs. NLRP3+/+ mice injected with HFD-sEVs. Figure S6. Novel-miR-7 antagomir does not alter the changes of body weight, circulating lipid content and aminotransferase activities of NAFLD mice. Mice were fed with a MCD diet for 2 weeks or a HFD for 6 weeks, followed by administration of 1 μmol/kg novel miR-7 antagomir or negative control (NC) lipoprotein cholesterol (LDL-C) content, n = 6 per group. (E-F) Plasma levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), n = 6 per group. Data are expressed as the mean ± SEM. Statistics: One-way ANOVA, **P < 0.01 vs. the MCS plus NC antagomir group; &P < 0.05, &&P < 0.01 vs. the ND plus NC antagomir group; Ns: no significance, the MCD plus novel-miR-7 antagomir group were compared with the MCD plus NC antagomir group, and the HFD plus novel-miR-7 antagomir group were compared with the HFD plus NC antagomir group. Figure S7 Novel-miR-7 antagomir reduces the cardiac cytokine contents and ameliorates coronary microvascular hyperpermeability of NAFLD mice. Mice were fed with a MCD diet for 2 weeks or a HFD for 6 weeks, followed by administration of 1 μmol/kg novel miR-7 antagomir or negative control (NC) antagomir via the caudal vein once a week. (A-D) Cardiac IL-1β and HMGB1 contents were measured by ELISA kits, n = 5-6 per group. (E-F) Representative images and the summarized data of Evans blue concentrations in heart, n = 6 per group. Data are expressed as the mean ± SEM. Statistics: One-way ANOVA, **P < 0.01 vs. the MCS plus NC antagomir group; &P < 0.05, &&P < 0.01 vs. the ND plus NC antagomir group; #P < 0.05, ##P < 0.01 vs. the MCD plus NC antagomir group; $$P Figure S8. NAFLD hepatic sEVs promote pulmonary microvascular hyperpermeability via novel-miR-7. Mouse lung microvascular endothelial cells (MLVECs) were transfected with 100 nmol/L novel-miR-7 inhibitor or negative control (NC) inhibitor and incubated with or without 120 μg/mL NAFLD hepatic sEVs for 24 h. Mice were fed with a MCD diet for 2 weeks or a HFD for 6 weeks, followed by administration of 1 μmol/kg novel miR-7 antagomir or negative control (NC) antagomir via the caudal vein once a week. (A-B) Representative flow cytometry images of caspase-1 FLICA staining and the summarized data of the positive cells, n = 4 per group. The fold changes were obtained by calculating the ratio of the positive cells of the treated groups to the NC inhibitor group. (C) Relative permeability of the endothelial monolayer to FITC-dextran, n = 4 per group. (D-E) Representative images and the summarized data of Evans blue concentrations in lung, n = 6 per group. Data are expressed as the mean ± SEM. Statistics: One-way ANOVA, *P < 0.05, **P < 0.01 vs. the NC inhibitor group or the MCS plus NC antagomir group; &&P < 0.01 vs. the NC inhibitor group or the ND plus NC antagomir group; #P < 0.05, ##P < 0.01 vs. the NC inhibitor plus MCD-sEVs group or the MCD plus NC antagomir group; $P < 0.05, $$P < 0.01 vs. the NC inhibitor plus HFD-sEVs group or the HFD plus NC antagomir group. Figure S9. Yield and size changes of hepatic sEVs between the two models. NAFLD or control hepatic sEVs were isolated by differential ultracentrifugation. (A) Protein concentration of hepatic sEVs was measured by BCA kit, n = 7–8 per group. (B) Representative images of sEV size distribution analysis by Flow Nanoanlyzer, n = 8 per group. (C) Concentration of hepatic sEVs was measured by Flow Nanoanlyzer, n = 8 per group. (D) Average size of hepatic sEVs was measured by Flow Nanoanlyzer, n = 8 per group. Data are expressed as the mean ±SEM. Statistics: One-way ANOVA, Ns: no significance, the MCD group were compared with the MCS group, and the HFD group were compared with the ND group. Figure S10. Genetic inhibition of NLRP3 suppresses NLRP3 inflammasome-associated microvascular hyperpermeability MVECs were transfected with 100 nmol/L NLRP3 siRNA (siNLRP3) or negative control siRNA (siNC) and incubated with or without 120 μg/mL NAFLD hepatic sEVs for 24 h. (A) Transfection efficiency of 5-FAM labeled siNLRP3. Scale bar, μm. (B) Representative western blot bands and the summarized data determined by densitometric analysis, n = 4 per group. (C-D) Representative flow cytometry images of caspase-1 FLICA staining and the summarized data of the positive cells, n = 4 per group. The fold changes were obtained by calculating the ratio of the positive cells of the treated groups to the siNC group. (E) Relative permeability of the endothelial monolayer to FITC-dextran, n = 4 per group. Data are expressed as the mean ± SEM. Statistics: One-way ANOVA, **P < 0.01 vs. siNC group; # P < 0.05, ## P < 0.01 vs. siNC plus MCD-sEVs group; &&P < 0.01 vs. siNC group; $$ P < 0.01 vs. siNC plus HFD-sEVs group. Figure S11. Effect of novel-miR-705 and novel-miR-1135 inhibitor on NLRP3 inflammasome activation. MVECs were transfected with 100nmol/L novel-miR-705 inhibitor or novel-miR-1135 inhibitor or negative control (NC) inhibitor and incubated with or without 120 μg/mL NAFLD hepatic sEVs for 24 h. (A-B) Representative flow cytometry images of caspase-1 FLICA staining and the summarized data of the positive cells, n = 4 per group. The fold changes were obtained by calculating the ratio of the positive cells of the treated groups to the NC inhibitor group. Data are expressed as the mean ± SEM. Statistics: One-way ANOVA, **P < 0.01 vs. NC inhibitor group; # P < 0.05 vs. NC inhibitor plus MCD-sEVs group; &&P < 0.01 vs. NC inhibitor group; Ns: no significance vs. NC inhibitor plus MCD-sEVs or HFD-sEVs groups. Figure S12. KEGG pathway enrichment analysis of the strong candidate genes. The common genes from cytokine activity and cytokine receptor binding signaling pathway with correlation scores ≥4 were considered as strong candidate genes of microvascular hyperpermeability. KEGG pathway enrichment analysis was performed to reveal the relevant signaling pathway of these strong candidate genes. Figure S13. Three-Dimensional cell model from the Z-stack obtained by stimulated emission depletion (STED) microscopy. MVECs were pretreated with or without 20 μmol/L dynasore for 40 min, followed by incubation of 120 μg/mL DiI-labeled hepatic sEVs for 8 h. Representative fluorescent confocal images of DiI-labeled hepatic sEVs (red), wheat germ agglutinin (WGA, green) with DAPI (blue). The pellet derived from the ultracentrifugation of DiI alone was used as a vehicle control. Scale bar, 10 μm.

Additional file 2: Table S1. Oligonucleotides. The synthesized oligonucleotides are listed in Table S1.

A systematic investigation has been carried out on the palladium-catalyzed thiocarbonylation and related reactions of unsaturated substrates such as propargylic and allylic alcohols, 1,2- and 1,3-dienes, and conjugated enynes with thiols and carbon monoxide. It has been first demonstrated that the reaction of allylic alcohols with thiols and CO in the presence of catalytic quantities of Pd(OAc) 2 (3 mol%), triphenylphosphine (12 mol%), and p-TsOH (5 mol%) leads to a novel reaction, namely thiocarbonylation, to afford beta,gamma-usaturated thiol esters in good to excellent yields. Other catalyst systems such as Pd 2(dba)3·CHCl3/PPh3/p-TsOH, Pd(PPh 3)4/p-TsOH, and Pd(OAC)2/dppb/ p-TsOH are also effective for this transformation. The thiocarbonylation reaction is believed to proceed via a eta3-allylpalladium intermediate. The reaction occurs in a highly regioselective manner, at the least hindered allylic terminal carbon of the substrate, to give the products. The new carbonylation procedure was readily applied to a variety of allylic alcohols, and to both aromatic and aliphatic thiols. Palladium (0) as well as palladium (II) complexes with added phosphine ligands catalyze the thiocarbonylation of propargylic compounds. Depending on the reaction conditions employed, the reaction can afford mono- and di-thioesters or beta-thio-substituted gamma-lactones as the principal products in good yields, respectively. Thus, in the presence of 3 mol% of Pd(OAC)2, 5 mol% of p-TsOH, and 12 mol% of PPh3, propargylic alcohols react with thiols in THF under 400 psi of carbon monoxide at 100°C to give monothioesters in excellent yields. beta-Thio-alpha,beta-unsaturated-gamma-lactones can be prepared in high yields by the thiolactonization of propargylic alcohols using Pd(PPh3)4 as the catalyst and DME as the solvent. Propargylic mesylates undergo dithiocarbonylation when they are treated with catalytic amounts of Pd(PPh3)4. This reaction exclusively gives dithioesters under milder conditions (90°C, 400 psi). A series of mono- and di-substituted allenes undergo direct thiocarbonylation with thiols and carbon monoxide to form the corresponding unsaturated thioesters in 73--94% isolated yields. This reaction requires catalytic quantities of Pd(OAC)2 (3 mol%) and triphenylphosphine (12 mol%) in THF under an atmosphere of CO (400 psi) at 100°C for 48 h. The reaction is believed to proceed via pi-allylpalladium complex. The reaction exhibits high regioselectivity, in which the thiophenyl group adds to the less substituted double bond of allenes. (Abstract shortened by UMI.)

Additional file 1. Multilingual abstracts in the five official working languages of the United Nations.

Additional file 1. Multilingual abstracts in the five official working languages of the United Nations.

Photocatalysis enables the cascade reactions of indoles and CBr4 in MeOH through a C(sp(2) )H functionalization/methanolysis sequence. The title reaction provides an efficient access to indole 2- and 3-carboxylates in a single operation (no preinstallation of protecting as well as directing groups was required) with good yields under mild reaction conditions.

The reaction of 1,3-conjugated enynes bearing a terminal triple bond, e.g., H2C:CMeC≡CH, with thiols, e.g., PhSH, and CO in the presence of catalytic amt. of Pd(OAc)2 and 1,3-bis(diphenylphosphino)propane in THF at 110° gave 2-(phenylthiocarbonyl) 1,3-dienes, e.g., MeC(:CH2)C(:CH2)COSPh, in moderate to good yields. The thiocarbonylation takes place with high chemo- and regioselectivity, with the attack by the phenylthiocarbonyl group occurring exclusively at C-2 of the 1,3-conjugated enyne.