Simultaneous quantitative profiling of N-acyl-l-homoserine lactone and 2-alkyl-4(1H)-quinolone families of quorum-sensing signaling molecules using LC-MS/MS

An LC-MS/MS method, using positive mode electrospray ionization, for the simultaneous, quantitative and targeted profiling of the N-acyl-L-homoserine lactone (AHL) and 2-alkyl 4-(1H)-quinolone (AQ) families of bacterial quorum-sensing signaling molecules (QSSMs) is presented. This LC-MS/MS technique was applied to determine the relative molar ratios of AHLs and AQs produced by Pseudomonas aeruginosa and the consequences of mutating individual or multiple QSSM synthase genes (lasI, rhlI, pqsA) on AHL and AQ profiles and concentrations. The AHL profile of P. aeruginosa was dominated by N-butanoyl-L-homoserine lactone (C4-HSL) with lesser concentrations of N-hexanoyl-L-homoserine lactone (C6-HSL) and 3-oxo-substituted longer chain AHLs including N-(3-oxodecanoyl)-L-homoserine lactone (3-oxo-C10-HSL) and N-(3-oxododecanoyl)-L-homoserine lactone (3-oxo-C12-HSL). The AQ profile of P. aeruginosa comprised the C7 and C9 long alkyl chain AQs including 2-heptyl-4-hydroxyquinoline (HHQ), 2-nonyl-4-hydroxyquinoline, the "pseudomonas quinolone signal" (2-heptyl-3-hydroxy-4-quinolone) and the N-oxides, 2-heptyl-4-hydroxyquinoline N-oxide and 2-nonyl-4-hydroxyquinoline N-oxide. Application of the method showed significant effects of growth medium type on the ratio and the nature of the QSSMs synthesized and the dramatic effect of single, double and triple mutations in the P. aeruginosa QS synthase genes. The LC-MS/MS methodology is applicable in organisms where either or both AHL and AQ QSSMs are produced and can provide comprehensive profiles and concentrations from a single sample.     

Comprehensive profiling of <Emphasis Type="Italic">N</Emphasis>-acylhomoserine lactones produced by <Emphasis Type="Italic">Yersinia pseudotuberculosis</Emphasis> using liquid chromatography coupled to hybrid quadrupole–linear ion trap mass spectrometry

A method for the comprehensive profiling of the N-acylhomoserine lactone (AHL) family of bacterial quorum-sensing molecules is presented using liquid chromatography (LC) coupled to hybrid quadrupole–linear ion trap (QqQLIT) mass spectrometry. Information-dependent acquisition (IDA), using triggered combinations of triple-quadrupole and linear ion trap modes in the same LC-MS/MS run, was used to simultaneously screen, quantify and identify multiple AHLs in a single sample. This MS method uses common AHL fragment ions attributed to the homoserine moiety and the 3-oxo-, 3-hydroxy- or unsubstituted acyl side chains, to identify unknown AHLs in cell-free culture supernatants in an unbiased manner. This LC-MS technique was applied to determine the relative molar ratios of AHLs produced by Yersinia pseudotuberculosis and the consequences of inactivating by mutation either or both of the AHL synthase genes (ypsI and ytbI) on AHL profile and concentration. The Y. pseudotuberculosis wild type but not the ypsI ytbI double mutant produced at least 24 different AHLs with acyl chains ranging from C4 to C15 with or without 3-oxo or 3-hydroxy substituents. YtbI, in contrast to YpsI, could direct the synthesis of all of the AHLs identified. The most abundant and hence most biologically relevant Y. pseudotuberculosis AHLs were found to be the 3-oxo-substituted C6, C7 and C8 AHLs and the unsubstituted C6 and C8 compounds. The LC-QqQLIT methodology is broadly applicable to quorum-sensing signal molecule analysis and can provide comprehensive AHL profiles and concentrations from a single sample and simultaneously collect confirmatory spectra for each AHL identified. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users. Catharine A. Ortori and Steve Atkinson made an equal contribution to the paper.     

Identification of bacterial <Emphasis Type="Italic">N</Emphasis>-acylhomoserine lactones (AHLs) with a combination of ultra-performance liquid chromatography (UPLC), ultra-high-resolution mass spectrometry,and in-situ biosensors

N-Acylated homoserine lactones (AHLs) are produced by Gram-negative bacteria as communication signals and are frequently studied as mediators of the “quorum sensing” response of bacterial communities. Several reports have recently been published on the identification of AHLs from different species and attempts have been made to study their role in natural habitats, for example the surface of plant roots in the rhizosphere. In this article, different analytical methods, including bacterial biosensors and chromatographic techniques, are reviewed. A concept for assignment of the structures of AHLs is also presented. The retention behaviour of derivatives of AHLs containing β-keto or hydroxyl groups and/or double bonds has been evaluated in relation to the separation behaviour of AHLs with saturated and unsubstituted alkanoyl chains. Samples have also been analysed by high resolution mass spectrometry (Fourier-transform ion-cyclotron-resonance mass spectrometry, FTICR-MS), nano liquid chromatography–electrospray ionization ion trap mass spectrometry (nano-LC–MS) and by the aid of a biosensor. The results obtained from ultra performance liquid chromatography (UPLC), FTICR-MS, nano-LC–MS, and bioassays have been compared to attempt structural characterisation of AHL without chemical synthesis of analytical standards. The method was used to identify the major AHL compound produced by the rhizosphere bacterium Acidovorax sp. N35 as N-(3-hydroxydecanoyl)homoserine lactone.     

Profiling of N-acyl-homoserine lactones by liquid chromatography coupled with electrospray ionization and a hybrid quadrupole linear ion-trap and Fourier-transform ion-cyclotron-resonance mass spectrometry (LC-ESI-LTQ-FTICR-MS)

A method for the comprehensive profiling of the N-acyl-homoserine lactone (AHL) family of bacterial quorum-sensing molecules is presented using liquid chromatography (LC) coupled to a hybrid quadrupole linear ion trap (LTQ) and Fourier-transform ion-cyclotron-resonance mass spectrometer (FTICR). We demonstrate an increase in signal intensity in MS with electrospray ionization (ESI) of the protonated molecules, [M + H](+), by using acetonitrile (ACN) instead of methanol (MeOH) as the organic solvent under the conditions in which the samples were supplied to the probe by direct infusion at constant flow rates. The presence of ACN prevents the formation of methanol adducts such as [M + MeOH + H](+) and [M + MeOH + Na](+), while also lowering the signal intensity of sodiated [M + Na](+) ions. Sensitivity of these signaling molecules in terms of signal-to-noise ratio (S/N) using low-resolution LTQ-MS and high-resolution FTICR-MS were compared under reversed-phase (RP) LC separations with ESI interface. Special emphasis was paid to the choice of the separation column, its elution conditions and detection of the major AHL compounds produced by the Serratia liquefaciens strain ATCC 27592. The most promising results were obtained using a RP C16-amide column eluted with a linear mobile phase gradient ACN/H(2)O containing 0.1% formic acid. The whole set of AHL homologs in bacterial extracts was detected in the extracted-ion chromatographic (XIC) mode, and the calculations of molecular formulae were performed by including the isotopic pattern. This mode of displaying data, with a very narrow mass-to-charge ratio window (i.e. +/- 0.0010 as m/z unit) around each selected ion, has allowed the identification of all the eight known homoserine lactones, viz. C(4)-HSL, 3-oxo-C(6)-HSL, C(6)-HSL, 3-oxo-C(8)-HSL, C(8)-HSL, C(10)-HSL, C(12)-HSL and C(14)-HSL. In addition, at least four uncommon signaling mediators previously unreported, namely, 3-oxo-C(10:1)-HSL, 3-oxo-C(11:2)-HSL, 3-oxo-C(13:2)-HSL and 3-OH-C(16)-HSL, were identified and characterized; their roles in cell-to-cell communication has to be elucidated.     

Use of solid-phase extraction to enable enhanced detection of acyl homoserine lactones (AHLs) in environmental samples

A challenge for understanding the role of bacterial cell–cell signalling in the environment is the detection of those signals, which are often present in low (nmol L⁻¹) concentrations. We describe here a simple purification method, solid-phase extraction (SPE), for increasing the sensitivity of detection for one such group of signals, acyl homoserine lactones (AHLs), in environmental samples. Spiking of dried marine sponge tissue (Stylinos sp.) with AHLs resulted in detection down to 0.01 ppm for 3-oxo-hexanoyl homoserine lactone (3-oxo C₆-HSL) and 1 ppm for hexanoyl homoserine lactone (C₆-HSL). Compared with liquid extraction methods use of SPE resulted in twofold and tenfold improvements in sensitivity, respectively.     

Robust routes for the synthesis of N-acylated-l-homoserine lactone (AHL) quorum sensing molecules with high levels of enantiomeric purity

The ready availability of native quorum sensing molecules and related structural analogues is of significant biological interest in the development of methods to manipulate bacterial quorum sensing systems in a useful fashion. In this Letter we report robust routes for the synthesis of a range of N-acylated-l-homoserine lactone (AHL) quorum sensing molecules. Crucially, we have analysed the enantiopurity of the final AHLs and in all cases, excellent levels were observed.     

Detection of bacterial quorum sensing <Emphasis Type="Italic">N</Emphasis>-acyl homoserine lactones in clinical samples

Bacteria communicate among themselves using certain chemical signaling molecules. These signaling molecules generally are N-acyl homoserine lactones (AHLs) in Gram-negative bacteria and oligopeptides in Gram-positive bacteria. In addition, both Gram-positive and Gram-negative bacteria produce a family of signaling molecules known as autoinducer-2 that they employ for their communications. Bacteria coordinate their behavior by releasing and responding to the chemical signaling molecules present in proportion to their population density. This phenomenon is known as quorum sensing. The role of bacteria in the pathogenesis of several diseases, including gastrointestinal (GI) disorders, is well established. Moreover, rather recently bacterial quorum sensing has been implicated in the onset of bacterial pathogenicity. Thus, we hypothesized that the signaling molecules involved in bacterial communication may serve as potential biomarkers for the diagnosis and management of several bacteria-related diseases. For that, we previously developed a method based on genetically engineered whole-cell sensing systems for the rapid, sensitive, cost-effective and quantitative detection of AHLs in biological samples, such as saliva and stool, from both healthy and diseased individuals with GI disorders. Although various analytical methods, based on physical-chemical techniques and bacterial whole-cell biosensors, have been developed for the detection of AHLs in the supernatants of bacterial cultures, only a few of them have been applied to AHL monitoring in real samples. In this paper, we report work performed in our laboratory and review that from others that describes the detection of AHLs in biological, clinical samples, and report some of our recent experimental results.     

(Z)-N-(4-Decenoyl)homoserine lactone, a new quorum-sensing molecule, produced by endobacteria associated with Mortierella alpina A-178

N-Acylhomoserine lactones (AHLs) are the conserved quorum-sensing signal molecules in Gram-negative bacteria. (Z)-N-(4-Decenoyl)homoserine lactone (1), a new AHL, was isolated from the culture broth of the fungus Mortierella alpina A-178 harboring bacterial endosymbionts, called endobacteria. The structure and absolute configuration of 1 were elucidated by EI-MS, chemical synthesis, and chiral GC analysis. The compound induced the expression of a QS-regulated reporter gene in Agrobacterium tumefaciens NTL4, although its activity was lower than that of N-decanoylhomoserine lactone (6).     

Kinetics and equilibrium studies of adsorption of chromium(VI) ion from industrial wastewater using Chrysophyllum albidum (Sapotaceae) seed shells

A new biosorbent has been prepared by coating Chrysophyllum albidum (Sapotaceae) seed shells with chitosan and/or oxidizing agents such as sulfuric acid. This study investigated the technical feasibility of activated and modified activated C. albidum seed shells carbons for the adsorption of chromium(VI) from aqueous solution. The sorption process with respect to its equilibria and kinetics as well as the effects of pH, contact time, adsorbent mass, adsorbate concentration and particle size on adsorption was also studied. The most effective pH range was found to be between 4.5 and 5 for the sorption of the metal ion. The pseudo-first-order rate equation by Lagergren and pseudo-second-order rate equation were tested on the kinetic data, the adsorption process followed pseudo-second-order rate kinetics, also, isotherm data was analyzed for possible agreement with the Langmuir and Freundlich adsorption isotherms, the Freundlich and Langmuir models for dynamics of metal ion uptake proposed in this work fitted the experimental data reasonably well. However, equilibrium sorption data were better represented by Langmuir model than Freundlich. The adsorption capacity calculated from Langmuir isotherm was 84.31, 76.23 and 59.63 mg Cr(VI)/g at initial pH of 3.0 at 30 °C for the particle size of 1.00–1.25 mm with the use of 12.5, 16.5 and 2.1 g/L of CACASC, CCASC and ACASC adsorbent mass, respectively. This readily available adsorbent is efficient in the uptake of Cr(VI) ion in aqueous solution, thus, it could be an excellent alternative for the removal of heavy metals and organic matter from water and wastewater.     

Deuterium-labelled N-acyl-L-homoserine lactones (AHLs)--inter-kingdom signalling molecules--synthesis, structural studies, and interactions with model lipid membranes

N-Acyl-l-homoserine lactones (AHLs) are synthesized by Gram-negative bacteria. These quorum-sensing molecules play an important role in the context of bacterial infection and biofilm formation. They also allow communication between microorganisms and eukaryotic cells (inter-kingdom signalling). However, very little is known about the entire mechanism of those interactions. Precise structural studies are required to analyse the different AHL isomers as only one form is biologically most active. Theoretical studies combined with experimental infrared and Raman spectroscopic data are therefore undertaken to characterise the obtained compounds. To mimic interactions between AHL and cell membranes, we studied the insertion of AHL in supported lipid bilayers, using vibrational sum-frequency-generation spectroscopy. Deuterium-labelled AHLs were thus synthesized. Starting from readily available deuterated fatty acids, a two-step procedure towards deuterated N-acyl-l-homoserine lactones with varying chain lengths is described. This included the acylation of Meldrum’s acid followed by amidation. Additionally, the detailed analytical evaluation of the products is presented herein.     

Alkaloid production by callous tissue cultures of Cereus peruvianus (Cactaceae)

The morphologically undifferentiated cells of nonregenerant callous tissue of Cereus peruvianus cultured in the original medium and in medium supplemented with tyrosine were used as an alkaloid source. Comparison of alkaloid production by C. peruvianus plants and by callous tissues indicated that alkaloid levels were almost twice as high in callous tissues as in shoots of C. peruvianus plants. The ratio of alkaloid concentration between mature plant and morphologically und ifferentiated cells of callous tissue was 1∶1.7. A relationship between culture medium containing tyrosine and alkaloid production was also observed in the callous tissues of C. peruvianus. Since increased alkaloid production may be induced by additional factors such as tyrosine, increasing levels of tyrosine or other conditions of the culture medium may be considered factors for inducing higher alkaloid production by C. peruvianus callous tissues.     

HPLC-MS法检测N-酰基-高丝氨酸内酯类信号分子

建立了HPLC-MS检测群体感应信号分子N-酰基-高丝氨酸内酯类(N-acyl-homoserine lactones,AHLs)化合物的方法。利用该方法对细菌发酵液中的AHLs进行分析,结果表明,样品只需简单的处理,在没有标样的条件下,可以对样品中任意的AHLs分子进行定性分析。该方法简单、高效,检测的灵敏度明显大于经典的TLC-B iosensor检测AHLs的方法,为从复杂混合物中鉴定AHLs提供了有效手段。     

Analysis of the explosive 2,4,6-trinitrophenylmethylnitramine (tetryl) in bush bean plants

Previous attempts to delineate the metabolism of the explosive 2,4,6-trinitrophenylmethylnitramine (tetryl) in plants have been unsuccessful. Development of an appropriate analytical methodology has been thwarted due to the extreme thermal and base lability of tetryl as well as its propensity to undergo photodecomposition. This study presents a methodology based on solvent extraction of plant tissue followed by fractionation of the organic extract on silica gel with subsequent determination of tetryl by HPLC. This methodology allowed 82.70 ± 5.54% recovery of tetryl from fortified bush bean leaves. The developed methodology was applied to study tetryl uptake and metabolism in bush bean plants exposed to tetryl-amended hydroponic cultures.     

Study of the influence of carboxymethylcellulose on the absorption of selenium (and selected metals) in a target plant

The aim of the present paper is the study of the influence of a polysaccharide (carboxymethylcellulose, CMC) on uptake by a target plant Lactuca sativa (LS) of selenium and some metals. LS was grown on a well characterized soil: such as, treated with 1.5 mg kg⁻¹ Se(IV) only and with two levels of CMC (3 and 30 mg kg⁻¹). Similar experiments were carried out by using Se(VI) instead of Se(IV). Uptake was evaluated through the quantification of total content of Se in dried leaves and roots by a suitable technique (graphite furnace atomic absorption spectrophotometry, instrumental neutron activation analysis and differential pulse cathodic stripping voltammetry). Results evidenced as the uptake of selenium was dependent on the form of selenium added to the soil: Se(VI) is accumulated much more then Se(IV) according to its lower toxicity and higher mobility. The simultaneous presence of CMC led to a lower selenium uptake in leaves, whereas no clear influence was evidenced in roots. Furthermore, the presence of CMC influenced also the mobility process (soil→plant) of several other metals: a lower content of them was detected in plants when CMC was present in the soil.     

Pyrolytic behaviour of different biomasses (angiosperms) (maize plants,straws, and wood) in low temperature pyrolysis

The thermal degradation of agricultural products and by-products (two kinds of maize plants, wheat, and barley straw) has been investigated by means of thermogravimetric/mass spectrometric analysis at heating rates from 1 to 10 °C/min. Large differences were found in the pyrolytic behaviour of the untreated samples, mainly caused by the high content of inorganics (ash content of about 4–6 wt%). These differences could be reduced by washing the samples with cold water. A kinetic model based on the formal kinetic parameters for the pyrolysis of the main components (hemicelluloses, lignin, and cellulose) and their degradable amounts was applied. To reduce the complexity of the model, only largely ash reduced samples were used. The formal kinetic parameters for the main components of barley straw and Gavott were individually determined. Although, different monomeric lignin degradation products were found for the angiosperms of grassy biomass in comparison to woody biomass, the formal kinetic parameters for lignin degradation are similar. The transferability of the formal kinetic parameters was successfully tested by applying them to a different straw type (wheat) and to a different maize cultivar (Doge) using the results of the biochemical analysis for the main components (hemicelluloses, lignin, and cellulose).     

Carbon dioxide adsorption by physisorption and chemisorption interactions in piperazine-grafted Ni(2)(dobdc) (dobdc = 1,4-dioxido-2,5-benzenedicarboxylate)

The metal-organic framework Ni(2)(dobdc) (CPO-27-Ni, where dobdc = 1,4-dioxido-2,5-benzenedicarboxylate) has been post-synthetically modified with piperazine (pip) - a known 'accelerator' to improve the kinetics of CO(2) uptake in alkanolamine solvents for chemical absorption - and the impact of the modification on the CO(2) uptake and selectivity over N(2) has been probed. While the modified framework, Ni(2)(dobdc)(pip)(0.5) (pip-CPO-27-Ni), exhibits a lower uptake of CO(2) compared with the non-grafted material, the selectivity for CO(2) over N(2) at 25 °C and at pressures pertinent to post-combustion flue gas capture (0.1-0.15 bar) is enhanced. Mechanistically, the interaction between the CO(2) molecules and the free amine sites in pip-CPO-27-Ni occurs via physisorption and chemisorption interactions, in which CO(2) binds to the framework with an isosteric heat of adsorption (-Q(st)) of 40.5 kJ mol(-1) at very low coverage (P = 0.033 mbar), followed by binding at a higher heat of adsorption (-Q(st) = 46.2 kJ mol(-1) at P = 3.55 mbar). Pure water adsorption isotherms revealed a two-step mechanism for uptake in CPO-27-Ni, consistent with adsorption into the first and second hydration spheres of Ni(2+) followed by subsequent uptake via physisorption into the pores. Additional steric hindrance in pip-CPO-27-Ni results in a single step only. The working capacity over multiple cycles was also investigated using a temperature swing adsorption process which revealed reversible CO(2) adsorption and desorption of 10 wt% over 10 cycles.     

Biosorption of nickel(II) and copper(II) ions from aqueous solution by Streptomyces coelicolor A3(2)

The biosorption of nickel(II) and copper(II) ions from aqueous solution by dried Streptomyces coelicolor A3(2) was studied as a function of concentration, pH and temperature. The optimum pH range for nickel and copper uptake was 8.0 and 5.0, respectively. At the optimal conditions, metal ion uptake was increased as the initial metal ion concentration increased up to 250 mg l(-1). At 250 mg l(-1) copper(II) ion uptake was 21.8% whereas nickel(II) ion uptake was found to be as high as 7.3% compared to those reported earlier in the literature. Metal ion uptake experiments were carried out at different temperatures where the best ion uptake was found to be at 25 degrees C. The characteristics of the adsorption process were investigated using Scatchard analysis at 25 degrees C. Scatchard analysis of the equilibrium binding data for metal ions on S. coelicolor A3(2) gave rise to a linear plot, indicating that the Langmuir model could be applied. However, for nickel(II) ion, divergence from the Scatchard plot was evident, consistent with the participation of secondary equilibrium effects in the adsorption process. Adsorption behaviour of nickel(II) and copper(II) ions on the S. coelicolor A3(2) can be expressed by both the Langmuir and Freundlich isotherms. The adsorption data with respect to both metals provide an excellent fit to the Freundlich isotherm. However, when the Langmuir isotherm model was applied to these data, a good fit was obtained for the copper adsorption only and not for nickel(II) ion.     

Biosorption of Pb(II) and Cd(II) ions by Agave sisalana (sisal fiber)

The present work proposes the use of Agave sisalana (sisal fiber) as an natural adsorbent for ions Pb(II) and Cd(II) biosorption from natural waters. The flame atomic absorption spectrometry was used for quantitative determination and study of the ions Pb(II) and Cd(II) adsorption on the solid phase. The Fourier transform infrared spectroscopy (FT IR) was used to investigate the sisal structure and the specific BET surface area was analyzed. The biosorption potential of sisal as biosorbent for the removal of the ions Pb(II) and Cd(II) from aqueous solution was investigate considering the followings parameters: pH, biomass amount and contact time. Langmuir and Freundlich isotherms were used to evaluate adsorption behavior of the ions on this solid phase. The results showed that sisal has a surface area to adsorption of 0.0233 m² g^{− 1}, and the OH and CO functional groups are the main involved in the biosorption. The best interpretation for the experimental data was given by Freundlich isotherm that proposes a monolayer sorption with a heterogeneous energetic distribution of active sites, accompanied by interactions between sorbed molecules. The maximum monolayer biosorption capacity was found to be 1.85 mg g^{− 1} for Cd (II) and 1.34 mg g^{− 1} for Pb (II) at pH 7 and 296 K. This phase solid can be used for biosorption of cadmium and lead in polluted natural waters.     

Fast microwave‐assisted extraction of rotenone for its quantification in seeds of yam bean (Pachyrhizus sp.)

The aim of this study was to find if fast microwave‐assisted extraction could be an alternative to the conventional Soxhlet extraction for the quantification of rotenone in yam bean seeds by SPE and HPLC‐UV. For this purpose, an experimental design was used to determine the optimal conditions of the microwave extraction. Then the values of the quantification on three accessions from two different species of yam bean seeds were compared using the two different kinds of extraction. A microwave extraction of 11 min at 55°C using methanol/dichloromethane (50:50) allowed rotenone extraction either equivalently or more efficiently than the 8‐h‐Soxhlet extraction method and was less sensitive to moisture content. The selectivity, precision, trueness, accuracy, and limit of quantification of the method with microwave extraction were also demonstrated.     

Effect of strontium substitution on the activity of La2?xSrxNiO4 (x = 0.0–1.2) in NO decomposition

The aim of this work is to study the effect of Sr substitution on the redox properties and catalytic activity of La_2−x Sr _x NiO₄ (x = 0.0–1.2) for NO decomposition. Results suggest that the x = 0.6 sample shows the highest activity. The characterization (TPD, TPR, etc.) of samples indicates that the x = 0.6 sample possesses suitable abilities in both oxidation and reduction, which facilitates the proceeding of oxygen desorption and NO adsorption. At temperature below 700°C, the oxygen desorption is difficult, and is the rate-determining step of NO decomposition. With the increase of reaction temperature (T > 700°C), the oxygen desorption is favorable and, the active adsorption of NO on the active site (NO + V _o + Ni²⁺ → NO⁻-Ni³⁺) turns out to be the rate-determining step. The existence of oxygen vacancy is the prerequisite condition for NO decomposition, but its quantity does not relate much to the activity. Supported by the National Hi-Tech Research and Development Program of China (863 Program)(Grant No. 2004CB 719502) and the National Natural Science Foundation of China (Grant No. 20177022)