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A biphasic chiral recognition system based on chiral ligand exchange with Cu(II)- N- n-dodecyl-L-proline and hydroxypropyl-β-cyclodextrin as an additive was developed to enantioseparate aromatic β-amino acids by high-speed counter-current chromatography. The biphasic chiral recognition system was established with an n-butanol/water (1:1, v/v) solvent system by adding N- n-dodecyl-L-proline and Cu(II) ions to the organic phase and hydroxypropyl-β-cyclodextrin to the aqueous phase. Several separation parameters, such as temperature, pH value, and chiral selector concentration, were systematically investigated by enantioselective liquid-liquid extraction. Under the optimal separation conditions, 54.5 mg of ( R, S)-β-phenylalanine and 74.3 mg of ( R, S)-β-3,4-dimethoxyphenylalanine were baseline enantioseparated. More importantly, the synergistic enantiorecognition mechanism, based on the Cu(II)- N- n-dodecyl-L-proline and hydroxypropyl-β-cyclodextrin, was discussed for the first time. [ABSTRACT FROM AUTHOR]

The use of a dinuclear Cu 2 (II)-β-cyclodextrin complex (Cu 2 -β-CyD) as a chiral selector (CS) was successfully applied to chiral high speed counter-current chromatography (HSCCC) for the enantioseparation of aromatic α-hydroxycarboxylic acids. α-cyclohexylmandelic acid (CHMA) as a case was studied to optimize the separation temperature, the pH value of the aqueous phase, and the concentration of the chiral selector, which were comparatively studied between Cu 2 -β-CyD and native β-cyclodextrin (β-CyD) as CSs. The thermodynamic parameters of the formation of the complex between the enantiomers and the CSs were obtained, and the complex formation constants between the enantiomers and Cu 2 -β-CyD or β-CyD were determined using induced circular dichroism technology. The results indicate that Cu 2 -β-CyD can remarkably improve the enantioseparation compared with β-CyD, mainly due to the ion-pairing interactions between the deprotonation of carboxyl group of CHMA and the Cu(II) ion of Cu 2 -β-CyD. Five of other aromatic α-hydroxycarboxylic acids were also enantioseparated successfully by HSCCC using Cu 2 -β-CyD as a CS. [ABSTRACT FROM AUTHOR]

Highlights: [•] Six partially acylated pentasaccharide resin glycosides were isolated from Ipomoea cairica. [•] The absolute configuration of the aglycone was established as 11S by Mosher’s method. [•] The monosaccharides of cairicosides A–F were proved by GC–MS. [•] The cytotoxic activities of the pentasaccharide resin glycosides were evaluated in vitro. [Copyright &y& Elsevier]

Glycosides including triterpenoid saponins and flavonoid glycosides are the main constituents of Glycyrrhiza uralensis Fisch (licorice) and exhibit prominent pharmacological activities. However, conventional methods for the separation of glycosides always cause irreversible adsorption and unavoidable loss of sample due to their high hydrophilicities. The present paper describes a convenient method for the simultaneous separation of triterpenoid saponins and flavonoid glycosides from licorice by pH-zone-refining counter-current chromatography. Ethyl acetate/ n-butanol/water (2:3:5, v/v) with 10 mM TFA in the upper organic stationary phase and 10 mM ammonia in the lower aqueous mobile phase was used as the biphasic solvent system. Three triterpenoid saponins and two flavonoid glycosides including licorice-saponin A3 (63.3 mg), glycyrrhizic acid (342.2 mg), 3- O-[ β- d-glucuronopyranosyl-(1 → 2)- β- d-galactopyranosyl]glycyrrhetic acid (56.0 mg), liquiritin apioside (232.6 mg), and liquiritin (386.5 mg) were successfully obtained from licorice ethanol extract (2 g) in one step. This method subtly takes advantage of the common acidic properties of triterpenoid saponins and flavonoid glycosides, and obviously is much more efficient and convenient than the previous methods. It is also the first time that the separation of acidic triterpenoid saponins by using p H-zone-refining counter-current chromatography has been reported. [ABSTRACT FROM AUTHOR]

Highlights: [•] Two triterpenoids, fourteen lignans and three phenolics were obtained from Boschniakia himalaica. [•] Fourteen of these compounds were reported for the first time from B. himalaica. [•] Compound 17 was reported as a natural product for the first time. [•] The presence of lignans and their glycosides will be chemotaxonomically significant for B. himalaica. [ABSTRACT FROM AUTHOR]

Abstract: Preparative separation of seven phenylpropenoid glycerides from the bulbs of Lilium lancifolium (lily), was conducted by high-speed counter-current chromatography (HSCCC) with a two-phase solvent system composed of n-hexane–ethyl acetate–methanol–0.05% aqueous trifluoroacetic acid (TFA) (3:5:3:5, v/v/v/v). These phenylpropenoid glycerides were identified as 1-O-feruloylglycerol (1), 1-O-p-coumaroylglycerol (2), 1-O-caffeoyl-3-O-p-coumaroylglycerol (3), 1,2-O-diferuloylglycerol (4), 1,3-O-diferuloylglycerol (5), 1-O-feruloyl-3-O-p-coumaroylglycerol (6), and 1,3-O-di-p-coumaroylglycerol (7), respectively, by ESI-MS, 1H NMR and 13C NMR. Their antioxidant activities were evaluated by DPPH radical scavenging activity, ABTS radical cation scavenging activity, and ferric-reducing antioxidant power (FRAP). The trend in antioxidant capacity was similar in all the three assays, with 3>1, 4, 5, 6>2, 7. This is the first report on simultaneous separation of seven antioxidantive phenylpropenoid glycerides from L. lancifolium by HSCCC. [Copyright &y& Elsevier]

Ginsenosides Re and Rg1, the main bioactive components of Panax ginseng, have been preparatively isolated and purified using high-speed counter-current chromatography (HSCCC) combined with MCI gel column chromatography (MCI gel CC). Guided by HPLC-MS, the crude extracts from the roots of P. ginseng were pre-separated on an MCI gel column to obtain mixtures of ginsenosides Re and Rg1. Then, a 200 mg volume of enriched sample was separated by HSCCC with a two-phase solvent system composed of ethyl acetate–n-butanol–water (4: 1: 6, v/v/v), yielding 52.6 mg of Rg1 and 37.2 mg of Re with the purities of 99.3% and 98.2%, respectively. The total recovery of Rg1 and Re was 82.4% and 84.4%, respectively, in the two-step separation. This study introduces a fast and efficient method for purification of ginsenosides Re and Rg1 from P. ginseng. [ABSTRACT FROM AUTHOR]

Abstract: A levan-type exopolysaccharide (EPS) from Paenibacillus polymyxa EJS-3 was successfully acetylated, phosphorylated and benzylated, respectively, affording its derivatives of acetylated levan (AL), phosphorylated levan (PL) and benzylated levan (BL). Then, the antioxidant and antitumor activities in vitro of the natural polysaccharide and its derivatives were determined. As results, AL, BL and PL all exhibited higher reducing power, scavenging activity against superoxide radical and scavenging activity on hydroxyl radical than the natural polysaccharide, EPS-1. In addition, AL, BL and PL also exhibited higher antiproliferative activity against human gastric cancer BGC-823 cells in vitro than EPS-1. The enhanced activities of the derivatives were probably due to the introduction of acetyl, benzyl, or phosphoryl groups into EPS-1 molecules that increased electron-donating ability and affinity with the receptors on immune cells. The results suggested that the derivatives could be explored as promising antioxidant and antitumor agents. [Copyright &y& Elsevier]

High-speed counter-current chromatography (HSCCC) coupled with evaporative light scattering detection (ELSD) was successfully applied for preparative separation and purification of four steroidal saponins (compounds 1–4) from crude extracts of garlic (Allium sativum L.) by one-step separation, using n-butanol-acetic acid-6 mM aqueous ammonium acetate (4:1:5, v/v/v) as a solvent system. The quantities of compounds 1–4 isolated from 300 mg of the crude sample were 23.5 mg, 14.7 mg, 18.2 mg, and 7.0 mg, respectively. Their purities were found to be over 93.0% by HPLC-ELSD analysis. Their chemical structures were confirmed by ESI-MS, 1H- and 13C- NMR. This is the first time steroidal saponins have been separated using HSCCC-ELSD. [ABSTRACT FROM PUBLISHER]

Abstract: A rare 16β-H steroidal alkaloid saponin (1), an avenacoside-type saponin (2), two steroidal saponins (4, 5), one revised-structure steroidal saponin (3) and six known compounds (6–11) were isolated from aerial parts of Solanum surattense Burm. f. Their structures were established on the basis of physical data, as well as by using spectroscopic (HRESIMS, 1D and 2D NMR), and chemical analysis methods. Compounds 1 and 11 showed cytotoxicity against A549 cell line with IC50 values of 20.3 and 15.7μM, respectively. [Copyright &y& Elsevier]

Abstract: A novel separation technique of complexation high-speed counter-current chromatography (HSCCC) using copper ion as a complexation agent was first developed to isolate 5-hydroxyisoflavone isomers from Belamcanda chinensis (L.) DC. According to the partition coefficient and separation factor, the two-phase solvent system composed of light petroleum–ethyl acetate–methanol–water (3:5:3:5, v/v) and copper nitrate (0.10mol/L in the lower phase) was selected. 9.2mg isoirigenin (1), 46.4mg irigenin (2) and 1.2mg 5,7,4′-trihydroxy-6,3′,5′-trimethoxyisoflavone (3) were simultaneously purified from 100mg crude extract by HSCCC with the purity of 95.06%, 96.98% and 93.69%, respectively. As evidenced by the results of UV–Vis spectroscopy, the stoichiometries of the copper ion with the three 5-hydroxyisoflavones were all 1:1 and their chelating power was 3 > 2 > 1. Those explained the complexation HSCCC behavior. It is the first report that includes the practical application of complexation HSCCC and explanation of its chromatographic behavior. [Copyright &y& Elsevier]

Abstract: The immunostimulatory activity of Hyriopsis cumingii polysaccharides (HCPS) was evaluated by using in vitro cell models and in vivo animal models. The results demonstrated that crude HCPS and its purified fractions (HCPS-1, HCPS-2 and HCPS-3) could promote the splenocyte proliferation, increase the activity of acid phosphatase in peritoneal macrophages and strengthen peritoneal macrophages to devour neutral red in vitro in a dose-dependent manner. Furthermore, HCPS-3 exhibited much stronger immunostimulatory activity in vitro than crude HCPS, HCPS-1 and HCPS-2, probably due to the higher sulfate content of HCPS-3. For assay in vivo, crude HCPS significantly increased the indices of spleen and thymus, the activity of lysozyme in serum and the swelling rate of earlap in delayed-type hypersensitivity in a dose-dependent manner. These results suggested that HCPS might be a potential natural immunomodulator. [ABSTRACT FROM AUTHOR]

Abstract: The antioxidant activities of exopolysaccharides (EPS) from endophytic bacterium Paenibacillus polymyxa EJS-3 were evaluated by various methods in vitro and in vivo. In antioxidant assays in vitro, both crude EPS and its purified fractions (EPS-1 and EPS-2) were found to have moderate 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, hydrogen peroxide scavenging activity, lipid peroxidation inhibition effect, and strong ferrous ion chelating activity. And the antioxidant activities in vitro of EPS decreased in the order of crude EPS>EPS-2>EPS-1. In antioxidant assays in vivo, mice were subcutaneously injected with d-galactose (d-Gal) for 6 weeks and administered EPS-1 via gavage simultaneously. As a result, administration of EPS-1 significantly increased the thymus and spleen indices of d-Gal induced aging mice. Moreover, EPS-1 administration significantly enhanced the activities of antioxidant enzymes and total antioxidant capacity and decreased the levels of malondialdehyde in both serums and livers of aging mice. These results suggested that EPS had potent antioxidant activities and could be explored as novel natural antioxidant. [Copyright &y& Elsevier]

Abstract: Two minor coumarins, including a new compound qianhucoumarin J, were isolated from Peucedanum praeruptorum by a multi-step separation procedure using high-speed counter-current chromatography (HSCCC) and preparative high performance liquid chromatography (prep-HPLC) monitored by high performance liquid chromatography coupled with mass spectrometry (HPLC/MS) analysis. The crude extract was analyzed first by HPLC/MS to detect unknown compounds. The target compounds were enriched by HSCCC, using a two-phase solvent system of petroleum ether–ethyl acetate–methanol–water (5:5:6:4, v/v) and then isolated and purified by preparative HPLC monitored by HPLC/MS. The structure of the new compound was elucidated mainly by analysis of its 1D and 2D NMR spectral data. Its absolute configuration was determined by chemical degradation followed by chiral HPLC analysis. [ABSTRACT FROM AUTHOR]

Abstract: A simple method for the simultaneous decoloration and deproteinization of crude levan extract from the endophytic bacterium Paenibacillus polymyxa EJS-3 was developed through static and dynamic adsorption tests of macroporous resins. S-8 resin demonstrated the highest decoloration and deproteinization ratios among various resins tested. Under optimized static adsorption conditions (pH 6.0, 35°C and adsorption time of 70min), the ratios of decoloration, deproteinization and polysaccharide recovery for S-8 resin were 76.8%, 78.9% and 69.0%, respectively. Under optimized dynamic adsorption condition (flow rate of 2BV/h, 160ml of 2.5mg/ml crude levan extract), higher ratios of decoloration, deproteinization and polysaccharide recovery for S-8 resin (84.6%, 91.7% and 81.3%, respectively) were observed. The method developed will provide a potential approach for large-scale production of levan from P. polymyxa EJS-3. [Copyright &y& Elsevier]

Abstract: Optimization of medium ingredients for the production of mycelial polysaccharides from Phellinus baumii Pilát (PBMP) in submerged culture, preliminary characterization and the evaluation of antioxidant activity in vivo of PBMP were carried out. An optimal medium for PBMP production was obtained through a 2(7−3) fractional factorial design and a central composite design in response surface methodology as follows (g/l): glucose 35.36, yeast extract 2.88, peptone 2.73, MgSO4 1.47, KH2PO4 1, VB1 0.0075 and diammonium oxalate monohydrate 0.3. The resulting PBMP was composed of d-glucose, d-galactose, l-fucose, d-mannose, and l-rhamnose in a molar ratio of 12.74:1.39:1.00:1.92:0.22. It was demonstrated that the administration of PBMP could increase the activities of antioxidant enzymes, decrease the levels of malondialodehyde, and enhance total antioxidant capabilities in livers and serums of d-galactose-induced mice. The results suggested that PBMP had potent antioxidant activity and could be explored as novel natural antioxidant. [Copyright &y& Elsevier]

Abstract: In this study, response surface methodology was employed to optimize the medium compositions for the production of exopolysaccharides (EPS) from endophytic bacterium Paenibacillus polymyxa EJS-3. Firstly, fractional factorial design was applied to evaluate the effects of different components in the medium. It was found that sucrose, yeast extract and CaCl2 influenced significantly the production of EPS. Then, steepest ascent method and central composite design were used to optimize the concentrations of the three variables. As results, the optimal medium compositions were determined as following (g/L): sucrose 188.2, yeast extract 25.8, K2HPO4 5 and CaCl2 0.34, with a corresponding yield of 35.26g/L. In addition, both polysaccharide fractions (EPS-1 and EPS-2) from crude EPS were mainly composed of (2→6)-linked β-d-fructofuranosyl residues backbone with (2→1)-linked branches based on their structural characterization by FT-IR spectroscopy, methylation analysis and 13C NMR spectroscopy. [Copyright &y& Elsevier]

Abstract: Optimization of medium composition for the production of exopolysaccharides (EPS) from Phellinus baumii Pilát in submerged culture and the immuno-stimulating activity of EPS were carried out. Firstly, the medium components having significant effect on EPS production were screened out to be glucose, yeast extract and diammonium oxalate monohydrate by using a 2(7−3) fractional factorial design. Secondly, the concentrations of the three factors were optimized using central composite design in response surface methodology. As results, a quadratic model was found to fit for EPS production, and the optimal medium composition was determined as following (g/l): 34.12 glucose, 4 peptone, 5.01 yeast extract, 0.88 diammonium oxalate monohydrate, 0.75 MgSO4 and 1 KH2PO4 and 0.0075 thiamine (VB1). A yield of 2.363±0.04g/l for EPS was observed in verification experiment. Finally, EPS from P. baumii Pilát was found to have direct immuno-stimulating activity in vitro on splenocyte proliferative response and acid phosphatase activity in peritoneal macrophages in a dose-dependent manner. [Copyright &y& Elsevier]

Abstract: The production, characterization and antioxidant activities in vitro of exopolysaccharides (EPS) from endophytic bacterium Paenibacillus polymyxa EJS-3 were investigated. For EPS production, the preferable culture conditions were 24°C and pH 8 for 60h with sucrose and yeast extract as the carbon and nitrogen sources, respectively. Notably, sucrose concentration was the prominent factor, and the maximum yield of EPS (22.82g/L) was obtained at a sucrose concentration of 160g/L. The crude EPS was purified by chromatography of DEAE-52 and Sephadex G-100, affording EPS-1 and EPS-2 with molecular weights of 1.22×106 and 8.69×105 Da, respectively. They were composed of mannose, fructose and glucose in a molar ratio of 2.59:29.83:1 and 4.23:36.59:1, respectively. In addition, both crude and purified EPS showed strong scavenging activities on superoxide and hydroxyl radicals, and their antioxidant activities decreased in the order of crude EPS>EPS-2>EPS-1. [Copyright &y& Elsevier]

Abstract: Two novel C-22 steroidal lactone saponins, namely solanolactosides A, B (1, 2) and two new spirostanol glycosides, namely torvosides M, N (3, 4) were isolated from ethanol extract of aerial parts of Solanum torvum. Their structures were characterized as solanolide 6-O-[α-l-rhamnopyranosyl-(1→3)-O-β-d-quinovopyranoside] (1), solanolide 6-O-[β-d-xylopyranosyl-(1→3)-O-β-d-quinovopyranoside] (2), yamogenin 3-O-[β-d-glucopyranosyl-(1→6)-O-β-d-glucopyranoside] (3) and neochlorogenin 3-O-[β-d-glucopyranosyl-(1→6)-O-β-d-glucopyranoside] (4) on the basis of spectroscopic analysis. The cytotoxicities of the saponins (1–4) were evaluated in vitro against a panel of human cancer cell lines. Compounds 3 and 4 showed significant cytotoxic activity with the cell lines. [Copyright &y& Elsevier]

Abstract: Combined with preparative high-performance liquid chromatography, high-speed counter-current chromatography was employed for isolation and purification of saponins from Gypsophila paniculata L. n-Hexane–n-butanol–methanol–0.02% TFA (1:9:1:9, v/v) was employed as solvent system and 210nm was chosen as the wavelength of ultraviolet detection for the first time. The research tried to compare HSCCC with prep-HPLC, and further integrated their advantages to improve separation efficiency. Five known triterpene saponins were identified by 13C NMR and ESI-MS and their purities were all above 96%. The results demonstrated that adopted method was a feasible, economical and efficient technique for rapid preparative isolation of saponins. [Copyright &y& Elsevier]

• A new strategy for precise separation of lysine-specific demethylase 1 inhibitors • Multi-mode CCC separation guided by molecular docking and LC-MS/MS analysis • The PH-zone refining and recycling CCC separation finished by only one instrument • Compound 3 showed the potential ability of prohibiting tumor invasion and metastasis It is significant to precisely isolate potential active compounds from medicinal herbs containing multiple compounds. Herein, a new strategy for precise separation of lysine-specific demethylase 1 (LSD1) inhibitors from the rhizome of Corydalis yanhusuo (RCY) using counter-current chromatography (CCC) guided by molecular docking and liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) analysis was established. First, representative alkaloids from RCY were docked with LSD1 for screening active skeleton compounds. Simultaneously, the crude extract of RCY was preliminarily separated via pH-zone refining CCC. Subsequently, guided by LC-MS/MS analysis of the fragmentation pathways, three potential active fractions were obtained, followed by further online-storage and recycling CCC separation. Finally, three high-purity target quaternary alkaloids compound 3 (dehydrocorydaline) , 7 (coptisine) , and 8 (columbamine) were successfully isolated as a new class of potential natural LSD1 inhibitors by only one CCC instrument with multiple modes. Compound 3 , with the highest LSD1 inhibition ratio of 2.44 μ M, was tested for its ability to inhibit tumor invasion and metastasis in U2OS cells. Therefore, the CCC separation guided by virtual screening is a promising method for the targeted isolation of enzyme inhibitors from medicinal herbs. [ABSTRACT FROM AUTHOR]

Highlights • Chiral CCC was superior to HPLC for preparative separation of synephrine. • pH-zone-refining CCC allowed efficient preparative enantioseparation of synephrine. • pH-zone-refining CCC was suited for preparative resolution of β-amino alcohols. Abstract Synephrine, a chiral adrenergic agonist, exists as R - (more active) and S -enantiomers (less active) that are difficult to separate because of their alkalinity and high polarity. Herein, we show that although this enantioseparation can hardly be achieved by high-performance liquid chromatography, acceptable preparative separation can be realized using high-speed counter-current chromatography (HSCCC) and pH-zone-refining counter-current chromatography (CCC) under optimal conditions. Specifically, 23.0 mg of S -synephrine and 25.0 mg of R -synephrine were isolated from 60.0 mg of racemic synephrine by conventional HSCCC, while 70.0 mg of S -synephrine and 69.0 mg of R -synephrine were obtained from 200.0 mg of racemic synephrine by pH-zone-refining CCC. The pH-zone-refining CCC was identified as the most efficient enantioseparation technique for synephrine and can be applied for the preparative enantioseparation of other β-amino alcohols. [ABSTRACT FROM AUTHOR]

Sirtuin1 (SIRT1), an NAD+-dependent histone deacetylase, plays a crucial role in regulating molecular signaling pathways. Recently, inhibition of SIRT1 rather than its activation shows the therapeutic potential for central nervous system disorder, however, the discovered SIRT1 inhibitors remains limited. In this work, a dual recognition-based strategy was developed to screen SIRT1 inhibitors from natural resources in situ. This approach utilized a Ni-modified metal-organic framework (Ni@Tyr@UiO-66-NH 2) along with cell lysate containing an engineered His-tagged SIRT1 protein, eliminating the need for purified proteins, pure compounds, and protein immobilization. The high-performance Ni@Tyr@UiO-66-NH 2 was synthesized by modifying the surface of UiO-66-NH 2 with Ni2+ ions to specifically capture His-tagged SIRT1 while persevering its enzyme activity. By employing dual recognition, in which Ni@Tyr@UiO-66-NH 2 recognized SIRT1 and SIRT1 recognized its ligands, the process of identifying SIRT1 inhibitors from complex matrix was vastly streamlined. The developed method allowed the efficient discovery of 16 natural SIRT1 inhibitors from Chinese herbs. Among them, 6 compounds were fully characterized, and suffruticosol A was found to have an excellent IC 50 value of 0.95 ± 0.12 μM. Overall, an innovative dual recognition-based strategy was proposed to efficiently identify SIRT1 inhibitors in this study, offering scientific clues for the development of drugs targeting CNS disorders. [Display omitted] • A new Ni-modified MOF (Ni@Tyr@UiO-66-NH 2) was synthesized. • The MOF adsorbed His-tagged SIRT1 efficiently while maintaining the enzyme activity. • A dual recognition-based strategy was established based on the new MOF and His-tag. • 16 new natural SIRT1 inhibitors were identified in situ by applying the strategy. [ABSTRACT FROM AUTHOR]

Identifying medicinally relevant compounds from natural resources generally involves the tedious work of screening plants for the desired activity before capturing the bioactive molecules from them. In this work, we created a paper-based ligand fishing platform to vastly simplify the discovery process. This paper-based method exploits the enzymatic cascade reaction between α -glucosidase (GAA), glucose oxidase (GOx), and horseradish peroxidase (HRP), to simultaneously screen the plants and capture the GAA inhibitors from them. The designed test strip could capture ligands in tandem with screening the plants, and it features a very simply operation based on direct visual assessment. Multiple acylated flavonol glycosides from the leaves of Quercus variabilis Blume were newly found to possess GAA inhibitory activities, and they may be potential leads for new antidiabetic medications. Our study demonstrates the prospect of the newly discovered GAA ligands as potential bioactive ingredients as well as the utility of the paper-based ligand fishing method. [Display omitted] • Test strip (GDTS) built on enzyme cascade reaction and dopamine-paper was newly set. • GDTS can screen bioactive plants and concurrently capture α -glucosidase inhibitors. • Four plants were screened by GDTS as targets from fourteen plants. • Twenty-two inhibitors were acquired from target plants via GDTS. • Acylated flavonol glucosides in Quercus variabilis were new GAA inhibitors. [ABSTRACT FROM AUTHOR]

The aim of the study was to investigate the usefulness of metanephrine (MN) and normetanephrine (NMN) in bilateral simultaneous adrenal vein sampling (AVS) with and without ACTH stimulation. The study was conducted in a single referral center. Prospective recruited patients with PA were treated with AVS. The effects of cortisol, MN and NMN on selectivity catheterization were assessed and determined by lateralization. We enrolled 54 patients with PA who were treated with simultaneous bilateral AVS with ACTH. The Selectivity Index (SI) calculated by MN was higher than that calculated by other indicators (p<0.001), the catheterization success rate of MN at baseline was the same as that of cortisol after ACTH stimulation, and in lateralization diagnosis, MN was not inferior to cortisol. In conclusion, among the studied indicators, MN is the best index for determining the catheterization selectivity in AVS, especially in the absence of ACTH stimulation. [ABSTRACT FROM AUTHOR]

Chrysanthemum morifolium Ramat. is a commonly used Chinese herb and food homologous plant with traditional effects such as anti-inflammatory, antifebrile, antibacterial and antiviral. Photoaging is one of the main causes of accelerated skin aging. Chrysanthemum morifolium Ramat. has reported to alleviate photodamage. In this study, we investigated the protective effect of the extract of buds of Chrysanthemum morifolium Ramat. (CE) on UVB-induced photoaging and further mechanism. The extract of buds of chrysanthemum was analyzed by HPLC-Q-TOF-MS/MS. Antioxidant activity was assessed by DPPH and ABTS assay. Cell viability examined by cell counting kit-8 assay. The ROS level was detected by fluorescent probe DCFH-DA. Protein expression evaluated by Western blotting. The skin tissue investigated by immunohistochemistry. CE significantly reversed the decrease of cell viability that induced by UVB in HaCaT and HFF-1 cells. Further analysis showed that CE alleviated photoaging by inhibiting the expression of mitogen-activated protein kinase (MAPK) and activating the NF-E2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway to promote the expression of antioxidant enzymes. Moreover, CE effectively improved the reduced skin hydration, disordered collagen and thickening epidermis caused by UVB in mice. All results demonstrated that CE had therapeutic effect on UVB-induced photoaging and provided theoretical basis for its further developing as a natural functional product with anti-photoaging effect. [Display omitted] [ABSTRACT FROM AUTHOR]

• Physalin H treatment suppresses LPS-induced inflammation and oxidative stress. • Physalin H blocks the interaction between NRF2 and KEAP1 to inhibit the ubiquitination of NRF2. • Physalin H-mediated NRF2 nuclear translocation and activation plays a protective role in acute lung injury. Physalin H (PH), a withanolide isolated from Physalis angulata L. has been reported to have anti-inflammatory effect. However, its impact on acute lung injury (ALI) remains unexplored. In this study, we observed that PH significantly alleviated inflammation in LPS-stimulated macrophages by suppressing the release of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) and down-regulating the expression of the inflammation-related genes. RNA sequencing analysis revealed a significant up-regulation of the NRF2 pathway by PH. Further investigation elucidated that PH attenuated the ubiquitination of NRF2 by impeding the interaction between NRF2 and KEAP1, thereby facilitating NRF2 nuclear translocation and up-regulating the expression of target genes. Consequently, it regulated redox system and exerted anti-inflammatory effect. Consistently, PH also significantly alleviated pathological damage and inflammation in LPS-induced ALI mice model, which could be reversed by administration of an NRF2 inhibitor. Collectively, these results suggest that PH ameliorates ALI by activating the KEAP1/NRF2 pathway. These findings provide a foundation for further development of pH as a new anti-inflammatory agent for ALI therapy. [ABSTRACT FROM AUTHOR]

Ferritinophagy is a cellular process to release redox-active iron. Excessive activation of ferritinophagy ultimately results in ferroptosis characterized by ROS accumulation which plays important roles in the development and progression of cancer. Sinomenine, a main bioactive alkaloid from the traditional Chinese medicine Sinomenum acutum , inhibits the proliferation of cancer cells by promoting ROS production. Herein, new compounds were designed and synthesized through the stepwise optimization of sinomenine. Among them, D3-3 induced the production of lipid ROS, and significantly promoted colorectal cancer cells to release the ferrous ion in an autophagy-dependent manner. Moreover, D3-3 enhanced the interaction of FTH1-NCOA4, indicating the activation of ferritinophagy. In vivo experiments showed that D3-3 restrained tumor growth and promoted lipid peroxidation in the HCT-116 xenograft model. These findings demonstrated that D3-3 is an inducer of ferritinophagy, eventually triggering ferroptosis. Compound D3-3, as the first molecule to be definitively demonstrated to induce ferritinophagy, is worth further evaluation as a promising drug candidate in the treatment of colorectal cancer. [Display omitted] • Design and synthesis 28 sinomenine derivatives. • Compound D3-3 significantly induced ferroptosis in HCT-116 cells. • Compound D3-3 caused iron overload in an autophagy-dependent manner by enhancing the interaction of NCOA4 and FTH1. • Compound D3-3 was able to promote lipid peroxidation in colon cancer cells in vivo. [ABSTRACT FROM AUTHOR]

Covalent modification of Keap1 results in reducing ubiquitination and the accumulation of Nrf2, which subsequently initiates the transcription of cellular anti-oxidant and anti-inflammatory genes. Iso- seco -tanapartholide (IST), a sesquiterpene isolated from the traditional Chinese medicine Artemisia argyi , had been reported to possess NF-κB inhibitory activity. However, its deep anti-inflammatory effects and direct target have never been reported. Here we show that IST activated Nrf2 and increased its target gene expression. In particular, LPS-caused inflammation in vitro and in vivo was mitigated by IST-induced Nrf2 activation but aggravated by Nrf2 inhibition. Mechanically, IST targeted Keap1 proteins via alkylating its cysteine residues 151, 273, 288, and so on. Subsequently, the modifying agent IST was displaced by intermolecular sulfhydryl disulfide interchange to lead to a disulfide dimer of Keap1. The resulting conformational change of Keap1 liberated Nrf2 from sequestration and allowed it translocation to the nucleus to activate the transcriptional program. Further studies demonstrated that Keap1 dimer formation contributed to the anti-inflammatory effects of IST. Taken together, our findings reveal a new mechanism for Nrf2 activation and provide a potential lead compound to treat inflammatory diseases through targeting Keap1. [Display omitted] • IST exerts anti-inflammatory effects in vitro and in vivo. • IST activates Nrf2 and its target gene expression. • IST covalently modifies Keap1 and induces its oligomerization. • IST-induced Keap1 dimer is cross-linked by intermolecular disulfide bridges. • Keap1 dimer formation contributes to the anti-inflammatory effects of IST. [ABSTRACT FROM AUTHOR]

[Display omitted] Previously, we have reported a withanolide-type steroid, named tubocapsenolide A (TA), which shows potent anti-proliferative activity in several cancer cell lines. However, its inhibitory effect on the Janus kinase/signal transducer and activator of transcription 3 (JAK/STAT3) pathway and therapeutic potential on osteosarcoma have not been reported. In the present study, we aimed to investigate the effect and molecular mechanism of TA in osteosarcoma. The biological functions of TA in U2OS cells were investigated using colony formation, 5-ethynyl-20-deoxyuridine (EDU) staining, and cell cycle/apoptosis assays. The interaction between TA and Src homology 2 phosphatase 2 (SHP-2) was detected by enzyme activity and validated by target-identification methods such as drug affinity responsive target stability (DARTS), cellular thermal shift assay (CETSA), and biolayer interferometry (BLI). The in vivo anti-tumor efficacy of TA was analyzed in the xenograft tumor model. Western blotting analysis was performed to detect the protein expression levels. TA exhibited antitumor activity against osteosarcoma both in vitro and in vivo by regulating the JAK/STAT3 signaling pathway. Mechanically, TA interacted with SHP-2 directly and activated its phosphatase activity. Importantly, protein tyrosine phosphatase (PTP) inhibitor, SHP-2 inhibitor, and SHP-2 siRNA could reverse the inhibitory effect of TA on the JAK/STAT3 signaling pathway and restored the TA-induced cell death. TA activated the phosphatase activity of SHP-2, which resulted in the inhibition of the JAK/STAT3 pathway and contributed to the antitumor efficacy of TA. Collectively, these findings suggested that TA could serve as a novel therapeutic agent for the treatment of osteosarcoma. [ABSTRACT FROM AUTHOR]

Highlights: [•] Crude capsule polysaccharides (CP) were purified to afford CP-1, CP-2 and CP-3. [•] In vitro cell assay, CP showed strong immunostimulatory activities. [•] CP-1 had pyranose rings and α-configuration glycosidic bond residues. [•] CP-1 had one backbone chain and two branch chains in one repeating unit. [Copyright &y& Elsevier]

Background and Purpose: Liver fibrosis is one of the leading causes of morbidity and mortality worldwide but lacks any acceptable therapy. The transcription factor glioma‐associated oncogene homologue 1 (GLI1) is a potentially important therapeutic target in liver fibrosis. This study investigates the anti‐fibrotic activities and potential mechanisms of the phytochemical, physalin B. Experimental Approach Two mouse models (CCl4 challenge and bile duct ligation) were used to assess antifibrotic effects of physalin B in vivo. Mouse primary hepatic stellate cells (pHSCs) and human HSC line LX‐2 also served as in vitro liver fibrosis models. Liver fibrogenic genes, GLI1 and GLI1 downstream genes were examined using Western blot and quantitative real‐time PCR (qRT‐PCR). GLI1 acetylation and LAP2α–HDAC1 interaction were analysed by co‐immunoprecipitation. Key Results: In vivo, physalin B administration attenuated hepatic histopathological injury and collagen accumulation and decreased expression of fibrogenic genes. Physalin B dose‐dependently suppressed fibrotic marker expression in LX‐2 cells and mouse pHSCs. Mechanistic studies showed that physalin B inhibited GLI activity by non‐canonical Hedgehog signalling. Physalin B blocked formation of lamina‐associated polypeptide 2α (LAP2α)/histone deacetylase 1 (HDAC1) complexes, thus inhibiting HDAC1‐mediated GLI1 deacetylation. Physalin B up‐regulated acetylation of GLI1, down‐regulated expression of GLI1 and subsequently inhibited HSC activation. Conclusion and Implications: Physalin B exerted potent antifibrotic effects in vitro and in vivo by disrupting LAP2α/HDAC1 complexes, increasing GLI1 acetylation and inactivating GLI1. This indicates that the phytochemical physalin B may be a potential therapeutic candidate for the treatment of liver fibrosis. [ABSTRACT FROM AUTHOR]

The aim of the study was to investigate the significance and influence of adrenocorticotropic hormone (ACTH) stimulation in primary aldosteronism (PA) patients with simultaneous bilateral adrenal vein sampling (AVS). All patients diagnosed with PA underwent simultaneous bilateral AVS with ACTH. In 95 patients, the post-ACTH SI significantly increased (p<0.001), and it gradually decreased from t10–t30 after ACTH stimulation (p<0.001). The unsuccessful catheterization decreased after ACTH stimulation. Time points within 20 min after ACTH stimulation were better for sampling, and the selectivity did not increase over longer periods. According to lateralization before and after ACTH stimulation, the patients could be divided into 3 groups (U, unilateral; B, bilateral): U/U , U/B or B/U, and B/B. Compared with the U/U group, in the U/B or B/U and B/B groups, the lateralization index (LI) was lower both at baseline and after ACTH stimulation (p<0.0001), the contralateral index (CLI) was higher after ACTH stimulation (p<0.003), the serum potassium level was higher (p<0.001), and the carbon dioxide combining power (CO2CP) and base excess (BE) levels were lower. In conclusion, in simultaneous bilateral AVS, ACTH stimulation had significant effects on increasing the catheterization selectivity. Lateralization change was observed after stimulation. After ACTH stimulation, fewer patients could be diagnosed with lateralized PA. Patients with consistent lateralized PA showed a more serious phenotype. [ABSTRACT FROM AUTHOR]

[Display omitted] • GGT-activated fluorescent probe was developed for diagnosing drug-induced liver injury (DILI). • The probe affords advantages of light-up NIR fluorescence, and large Stokes shift. • The probe enables real-time and in vivo monitoring GTT, and evaluating the remediation of DILI treatment. Drug-induced liver injury (DILI), induced by overdose or chronic administration of drugs, has become the leading cause of acute liver failure. Therefore, an accurate diagnostic method for DILI is critical to improve treatment efficiency. The production of γ-glutamyltranspeptidase (GGT) is closely related to the progression of drug-induced hepatotoxicity. KL-Glu exhibits a prominent GGT-activated NIR fluorescence (734 nm) with a large Stokes shift (137 nm) and good sensitivity/selectivity, making it favorable for real-time detection of endogenous GGT activity. Using this probe, we evaluated the GGT up-regulation under the acetaminophen-induced liver injury model. Moreover, KL-Glu was successfully used to assess liver injury induced by the natural active ingredient triptolide and the effective amelioration upon treatment with N -acetyl cysteine (NAC) or Glutathione (GSH) in cells and in vivo by fluorescent trapping the fluctuation of GGT for the first time. Therefore, the fluorescent probe KL-Glu can be used as a potential tool to explore the function of GGT in the progression of DILI and for the early diagnosis and prognostic evaluation of DILI. [ABSTRACT FROM AUTHOR]

In order to exploit an abundant and cheap carbon source for poly(3-hydroxybutyrate) (PHB) production, the rice husk was pretreated with dilute sulfuric acid and steam flash-explosion to enhance the enzymatic saccharification. The yield of reducing sugars of pretreated rice husk hydrolysate came up to 266.5 mg/g rice husk, much higher than that of untreated rice husk hydrolysate (72.67 mg/g rice husk). This result indicated that the pretreatment can significantly enhance the yield of reducing sugars. Then the hydrolysate was used as a sole carbon source for PHB production by using Cupriavidus necator. Response surface method was used to optimize the fermentation conditions. Under optimum fermentation conditions (carbon source, 31.81 gL−1; nitrogen source, 1.8 gL−1; pH-value, 7.0; temperature, 27.1 °C), the PHB yield was 5.0 gL−1, which was in good agreement with the predicted value. • It is the first time to pretreat rice hull with dilute sulfuric acid and SFE. • Dilute sulfuric acid and SFE is an effective pretreatment method for rice hull. • A new and abundant carbon source for PHB production has been found. • Usage of rice hull hydrolysate for PHB production can protect environment. [ABSTRACT FROM AUTHOR]

In terms of ligand fishing, the amount and the relative activity recovery of enzymes immobilized on magnetic particles and nanoparticles are not preeminent. Therefore, the metal-organic framework (MOF) UiO-66-NH 2 was synthesized to immobilize the porcine pancreatic lipase (PPL) via precipitation-cross-linking, and the resulting novel biological matrices named PPL@MOF manifested high PPL loading capacity (98.31 mg/g) and relative activity recovery (104.4%). Moreover, the novel enzyme-MOF composite was applied to screen lipase inhibitors from Prunella vulgaris L. to enrich and improve the techniques of ligand fishing. As a result, 13 lipase ligands were obtained, and 12 compounds were determined by HPLC-Q-TOF-MS/MS. All of these ligands were further confirmed to be potential inhibitors through the verification of the activity assay and molecular docking. The proposed approach based on PPL@MOF was superior in terms of abundant protein loading capacity, high enzyme catalytic activity and easy preparation process. Taken together, our newly developed method provided a new platform for efficient discovering bioactive molecules from natural herbs. Image 1 • Pancreatic lipase was successful immobilized onto the metal-organic framework via the precipitation-cross-linking method. • PPL@MOF manifested high PPL loading capacity (98.31 mg) and relative activity recovery (104.4%). • 13 compounds were harvested from Prunella vulgaris efficiently and conveniently by the method of ligand fishing based PPL@MOF. • MOF-enzyme composite was first used for obtaining bioactive compounds directly from natural herbs. [ABSTRACT FROM AUTHOR]

Withangulatin A (WA) has been reported to exhibit potent antitumor activity. However, its possible mechanism and direct proteomic targets remain unknown. Herein we report the subcellular localization of WA by designing and synthesizing its fluorescent analogues with coumarin moieties. Furthermore, sarco/endoplasmic reticulum calcium-ATPase (SERCA)2 was identified as the potential target of WA for its antitumor activity by chemical proteomics. [ABSTRACT FROM AUTHOR]

Abstract γ-Glutamyltranspeptidase (GGT) is a cell surface-bound enzyme that is closely implicated in various physiological disorders such as tumor. Thus, an efficient method for monitoring GGT is extremely important for biological studies and disease diagnosis. Herein, a near-infrared fluorescent probe (TMN-Glu) has been developed for turn-on trapping of GGT activity in vitro and in vivo based on conjugating a dicyanoisophorone derivative fluorophore with a GGT activatable γ-glutamyl amide moiety. Advantages of the probe include near-infrared emission (658 nm), with large Stokes shift (213 nm), high specificity, high sensitivity (LOD = 0.024 U/L), low cytotoxicity and high imaging resolution, allowing for the real-time imaging endogenous GGT in living cells. Probe TMN-Glu was highly feasible to report on the GGT levels in different types of cells. Notably, we also demonstrated its applicability in real-time visualizing endogenous GGT in tumor-bearing nude mice with low background interference. These results indicated that the probe held great promise for real-time sensing and tracking GGT in complex biological systems, which would be useful for basic researches and clinical diagnosis of GGT-related diseases. Graphical abstract fx1 Highlights • An enzyme-activatable NIR fluorescent probe for γ-glutamyltranspeptidase (GGT) with large Stokes shift (213 nm). • The probe exhibited prominent features including high sensitivity, good biocompatibility, and low background interference. • The probe was capable of real-time tracking GGT activity in cells and tumor-bearing mice. [ABSTRACT FROM AUTHOR]

Ovarian cysts are one of the most common gynecologic affections for females. The most effective therapy is surgery, but not for all conditions. An 18‐year‐old woman was referred to our hospital because of menstruation disorder and abdominal distension. Ultrasound and computer tomography of the abdomen revealed a giant ovarian cyst. Magnetic resonance imaging revealed profound pituitary enlargement. Laboratory studies showed severe hypothyroidism, mild anemia, hyperlipidemia, hyperprolactinemia and an elevated level of cancer antigen‐125. Regression of the giant ovarian cyst and pituitary enlargement was observed after a 5‐month levothyroxine replacement therapy. Thus, for patients with ovarian cysts, hypothyroidism should be taken into account. Making correct diagnosis would avoid unnecessary surgery. [ABSTRACT FROM AUTHOR]

Countercurrent chromatography (CCC) has gradually become a widely used method for preparative separation of bioactive natural molecules. These molecules generally contain distinct scaffolds and characteristics that cannot be readily isolated from plants. While one-dimensional CCC is typically used for the initial purification with insufficiently resolved peaks after locating bioactive components, two-dimensional (2D) or multi-dimensional CCC strategies are employed to improve the resolution of peaks. However, these methods usually present certain disadvantages, such as complicated procedures and increased time consumption, experimental costs, and equipment requirements. Here, a bioactivity-guided cut CCC strategy was established to isolate lysine-specific demethylase 1 (LSD1) inhibitors from Scutellaria baicalensis Georgi. Gradient-elution CCC coupled with real-time detection of LSD1 inhibition by the collected fractions was developed. Next, an online-storage recycling CCC mode was designed to enable the active fractions to be stored in coils, and these active fractions were further separated to obtain pure compounds by using sequential recycling elution. In this strategy, active fractions are first identified, and then pure LSD1 inhibitors are isolated in the 2D CCC mode through continuous separation on a single instrument. By using our bioactivity-guided cut CCC strategy, we successfully isolated six natural LSD1 inhibitors from S. baicalensis Georgi, five of which were identified for the first time as natural LSD1 inhibitors. [ABSTRACT FROM AUTHOR]

A new method based on ligand fishing combined with high-performance liquid chromatography quadrupole-time-of-flight mass spectrometer and molecular docking was established to screen α -glucosidase inhibitors from a traditional Chinese medicine Morus alba root bark. α -Glucosidase was immobilized on magnetic nanoparticles, used as a solid support to incubate with crude extract. After ligand fishing, the eluates were analyzed by high-performance liquid chromatography quadrupole-time-of-flight mass spectrometer, obtaining eleven ligands ( 1 – 4 , 6 – 12 ) eventually. In order to discriminate the non-specific binders and discover powerful enzyme inhibitors, molecular docking was further performed and three of the eleven ligands were optimized to be excellent α -glucosidase inhibitors by the confirmation of isolation and bioassay of individual compounds. These three ligands, sanggenons G ( 6 ), O ( 7 ) and sanggenol G ( 12 ) exhibited striking inhibitory activities with extremely low IC 50 values. The results suggest that established method will be applied to a wide range of target protein to screen potential bioactive constituents from herbal medicines. [ABSTRACT FROM AUTHOR]

Cu(II) ion was selected as an additive to improve the enantioseparation efficiency of three dihydroflavone enantiomers in high-speed counter-current chromatography (HSCCC), using hydroxypropyl-β-cyclodextrin (HP-β-CyD) as the chiral selector. The influences of important parameters, including the metal ion, the concentrations of HP-β-CyD and the Cu(II) ion, and the sample size were investigated. Under optimal conditions, three dihydroflavone enantiomers, including (±)-hesperetin, (±)-naringenin, and (±)-farrerol, were successfully enantioseparated. The chiral recognition mechanism was investigated. The enantioseparation was attributed to the different thermodynamic stabilities of the binary complexes of HP-β-CyD and (±)-hesperetin, and Cu(II) ion could enhance this difference by forming ternary complexes with the binary complexes. This Cu(II) ion-improved complexation HSCCC system exhibited improved performance for chiral separation, and therefore it has great application potential in the preparative enantioseparation of other compounds with similar skeletons. [ABSTRACT FROM AUTHOR]

[Display omitted] • IST induces p62 to form covalent non-disulfide crosslinked oligomers. • Cysteine residues are necessary for IST-mediated p62 oligomerization. • p62 oligomerization promotes recruitment of KEAP1 for autophagic degradation. • IST-mediated p62/NRF2 activation plays a protective role in rheumatoid arthritis. • IST displays a reduced anti-inflammatory response in p62-knockdown cells. SQSTM1/p62 sequesters intracellular aberrant proteins and mediates their selective autophagic degradation. p62 oligomerization posttranslational modification enhances its sequestration function and positively regulates the KEAP1-NRF2 redox pathway. However, the regulation of p62 covalent oligomerization has yet been poorly characterized. Here, we identified a natural small-molecule sesquiterpene, Iso- seco -tanapartholide (IST) modified p62 cysteine residues, which induced p62 to form crosslinked oligomers between TBS and TBS or TBS and PB1 domains in a covalently non-disulfide-linked manner. Using LC-MS/MS analysis and complementary approaches, we revealed that Cys residues of p62 were necessary for IST-induced covalent oligomer. This oligomerization promoted p62 recruitment of KEAP1 for degradation by autophagosomes and released NRF2 to the nucleus to activate the expression of downstream genes with anti-oxidant and anti-inflammatory capacities. Accordingly, IST-mediated p62/NRF2 activation conferred protection from oxidative and inflammatory destruction of rheumatoid arthritis in vitro and in vivo. In contrast, p62-knockdown cells displayed a reduced anti-oxidant response and increased pro-inflammatory cytokine secretion in response to TNF-α stimulation. Hence, our findings uncover an unrecognized role of IST in the regulation of p62 oligomerization and provide a new strategy for the treatment of rheumatoid arthritis. [ABSTRACT FROM AUTHOR]

An off-on fluorescent probe (CuCIN), coumarin-naphthol linked by an imine group, for detecting Cu 2+ in DMSO-HEPES buffer (1:99, v/v) solution was successfully developed. Mechanism studies suggested that CuCIN firstly formed a complex with Cu 2+ in a 1:2 metal-to-ligand ratio, and then 60-fold fluorescence enhancement was observed when Cu 2+ promoted the imine hydrolysis to release the fluorescence compound 7-diethylaminocoumarin-3-aldehyde. This probe displayed desired properties such as high selectivity toward other metal ions, low detection limit (12.7 nM) and low cytotoxicity. Bio-imaging study revealed that the new probe CuCIN could be applied for imaging living cells with good cell permeability. We hope to apply this probe in the biomedical research field for the imaging of disease-relevant Cu 2+ . [ABSTRACT FROM AUTHOR]

In traditional Chinese folk medicine, Artemisia argyi H.Lév. & Vaniot (A. argyi) has been used for thousands of years, and it is clinically used to treat bronchitis and asthma. However, the mechanism of action of A. argyi on respiratory tract inflammation is not clear. Accumulating evidence that phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3 (PFKFB3) is actively expressed in inflammation. Here, we found that iso- seco -tanapartholide (IST), a sesquiterpene isolated from A. argyi , exhibited potent anti-inflammatory activity and significant inhibition of PFKFB3 expression. Therefore, we evaluated the effect of IST on airway inflammation and revealed its possible mechanisms. This study aimed to investigate the protective effect and possible mechanism of IST in lipopolysaccharide (LPS)-induced acute lung injury in mice. In vitro , RAW264.7 cells and BMDMs were stimulated with LPS, and the level of NO and inflammatory factors TNF-α, IL-1β, and IL-6 were detected by Griess reagent and ELISA, respectively. The effect of IST on the levels of PFKFB3 and its downstream proteins (p-STAT3, p-p65) in cells was assayed by western blotting. Lactate and glycolytic phenotypes were detected by lactate kit and Seahorse assay. In vivo , a mouse model of acute lung injury was induced by LPS, and the levels of inflammatory factors were measured by ELISA. Expression of PFKFB3 and its downstream proteins (p-STAT3, p-p65) in mouse alveolar macrophages by western blotting analysis. Lung permeability assessment by Evans Blue dye assay. H&E staining and Immunocytochemistry were used to observe the protection of IST against lung injury. IST significantly reduced LPS-induced expression of PFKFB3 and its downstream proteins (p-STAT3, p-p65). The inhibition of PFKFB3 has an impact on the glycolytic phenotype, such as a reduction in the rate of extracellular acidification (ECAR) and elevated lactate levels, and an increase in the rate of cellular oxygen consumption (OCR). Furthermore, IST inhibited LPS-induced NO release and increased the expression of pro-inflammatory factors TNF-α, IL-1β, and IL-6. In vivo , IST reduced pulmonary edema in LPS-induced acute lung injury, improved lung function, and reduced levels of inflammatory factors and lactate secretion. These results suggest that IST improves the characteristics of ALI by inhibiting the expression of the PFKFB3-mediated glycolytic pathway and may be a potential anti-inflammatory agent for inflammation-related lung diseases. [Display omitted] • The sesquiterpene IST isolated from Artemisia argyi exhibited significant anti-inflammatory activity.- • IST significantly inhibited the expression of PFKFB3 and its downstream proteins. • IST may reduce the occurrence of aerobic glycolysis by inhibiting the PFKFB3-mediated glycolytic pathway. • IST can alleviate LPS-induced acute lung injury and improve lung function in mice. [ABSTRACT FROM AUTHOR]

Cu(II) complexed amino acid ionic liquid, Cu(II)-[1-butyl-3-methylimidazolium][L-Pro] (Cu(II)-[BMIm][L-Pro]), was successfully adopted as chiral ligand to improve the enantioseparation efficiency in high speed counter-current chromatography (HSCCC). For the enantioseparation of intractable naringenin (NRG) racemic mixtures, Cu(II)-[BMIm][L-Pro] coupled with hydroxypropyl-β-cyclodextrin (HP-β-CD) was successfully applied as dual chiral selectors in HSCCC. The influence of important parameters, including the concentration of the chiral selectors, the pH value, and the temperature were investigated. Under optimal conditions, 4.5 mg of (+)-NRG and 4.1 mg of (−)-NRG were successfully separated from 10 mg NRG racemic mixtures with the purity of 98%. The chiral recognition mechanism of dual chiral selectors was illuminated by the UV–vis and NMR spectra, suggesting that the enantioseparation was upon the difference of the thermodynamic stability of the quaternary complexes of Cu(II), [BMIm][L-Pro], HP-β-CD, and NRG. The results illustrated that the developed HSCCC system, based on the synergistic mechanism of Cu(II)-[BMIm][L-Pro] and HP-β-CD, exhibited better performance on enantioseparation and had great application potential in preparative chiral separation of natural products. [ABSTRACT FROM AUTHOR]

Hainanolide (HN) is a norditerpenoid metabolite extract from Cephalotaxus fortunei Hook. f. C. fortunei Hook. f. is renowned for the active alkaloids, such as harringtonine (HT) and homoharringtonin (HTT), which have been clinically used to treat chronic myeloid leukemia. Nowadays, diterpenoids, another important metabolite, attracted the attention of chemists. Among them, Hainanolide (HN), a cephalotane-type diterpenoid, has been proven to possess potent antitumor activities. However, the underlying therapeutic mechanisms of HN in anti-tumor have not been investigated yet. Our present study demonstrated that HN inhibited HCT-116 and HCT-15 cell proliferation in a dose- and time-dependent manner. Further studies demonstrated that HN can induce G2/M phase arrest and alter the Cdc25C/Cdc2/CyclinB1 proteins. Western blot indicated that HN promoted apoptosis by up-regulating Bax and down-regulated Bcl-2. And the caspase-3 and caspase-9 activities of HCT-116 and HCT-15 cells were increased. Transcriptome analysis is used to reveal the possible mechanism. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses suggested the genes were mainly enriched in the MAPK signaling pathway. Certainly, HN activates MAPK signaling pathway. In vivo, HN prevented the AOM/DSS-induced tumorigenesis of colon cancer in C57BL/6 mice. Our study indicated that HN inhibits the progression of colon cancer cells by blocking the cell cycle, inducing apoptosis, and activating the MAPK pathway. This study provides a theoretical and experimental scientific basis for future investigations of the antitumor effects of HN against colon cancer. • HN inhibited the proliferation of colon cancer cells in vivo and in vitro. • HN arrested the cell cycle in the G2/M phase by altering the cell cycle-regulated proteins. • HN induced the apoptosis of colon cancer cells by increasing the activities of caspase-3 and caspase-9. • The activation of MAPK signaling pathways was involved in the cytotoxic effect of HN on colon cancer cells. [ABSTRACT FROM AUTHOR]

Oxidative stress plays a critical role in the pathogenesis of diabetic vascular complications. Trans-δ-viniferin (TVN), a polyphenolic compound, has recently attracted much attention as an antioxidant exhibiting a hypoglycemic potential. In the present study, we aimed at investigating the protective effect of TVN against high glucose-induced oxidative stress in human umbilical vein endothelial cells (HUVECs) and the potential mechanism involved. We found that TVN attenuated reactive oxygen species (ROS) production, increased catalase (CAT) activity and decreased malondialdehyde (MDA) levels to ameliorate cell survival induced by 35 mM glucose. Meanwhile, it inhibited high glucose-induced apoptosis by maintaining Ca2+and preserving mitochondrial membrane potential (MMP) levels. The immunoblot analysis indicated that TVN efficiently regulated the cleavage of caspase family, p53, Bax and Bcl-2, all mediated by SIRT1. Furthermore, the increased level of SIRT1 induced by TVN was inhibited by nicotinamide and siRNA-medicated SIRT1 silencing (si-SIRT1), thereby confirming the significant role of SIRT1 in these events. In conclusion, our results indicated that TVN efficiently reduced oxidative stress and maintained mitochondrial function related with activating SIRT1 in high glucose-treated HUVECs. It suggested that TVN is pharmacologically promising for treating diabetic cardiovascular complications. [ABSTRACT FROM AUTHOR]

The excessive expression of γ-glutamyltranspeptidase (GGT) is known to be closely associated with the progression, invasion and metastasis of malignant tumors, enabling GGT to be a cancer-related biomarker for cancer diagnosis. The development of the multifunctional fluorescent probe that combines GGT measurement and photodynamic therapy (PDT) into a single platform possesses exciting prospect for concurrent tumor diagnosis and treatment. In this study, we designed a new near-infrared (NIR) fluorescent probe (CyI-Glu) with therapeutic function which could realize the integration of fluorescence endogenous GGT visualization for tumor diagnosis and NIR imaging-guided tumor photodynamic therapy. CyI-Glu manifested a significant turn-on NIR fluorescence (718 nm) in response to GGT, which made it favorable for monitoring GGT activity with high selectivity and sensitivity (0.014 U/L) in vitro and in vivo. Furthermore, this GGT-responsive probe possessed improved ROS generation capacity for tumor cell apoptosis and tumor growth inhibition under NIR light irradiation. Collectively, the multifunctional fluorescent probe CyI-Glu with complementary advantages demonstrates significant potential for the diagnosis and treatment of tumors in complex organisms. [Display omitted] • Multifunctional fluorescent probe was developed for detecting GGT and photodynamic therapy of tumor. • The probe exhibited high selectivity/sensitivity for GGT analysis with distinct NIR fluorescence. • The probe could effectively inhibit tumor growth through generation of singlet oxygen upon NIR photoirradiation. [ABSTRACT FROM AUTHOR]