Discovery of 4-phenyl-1H-indazole derivatives as novel small-molecule inhibitors targeting the PD-1/PD-L1 interaction.

The inhibition of the programmed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) pathway with small molecules is a promising approach for cancer immunotherapy. Herein, novel small molecules compounds bearing various scaffolds including thiophene, thiazole, tetrahydroquinoline, benzimidazole and indazole were designed, synthesized and evaluated for their inhibitory activity against the PD-1/PD-L1 interaction. Among them, compound Z13 exhibited the most potent activity with IC50 of 189.6 nM in the homogeneous time-resolved fluorescence (HTRF) binding assay. Surface plasmon resonance (SPR) assay demonstrated that Z13 bound to PD-L1 with high affinity (KD values of 231 nM and 311 nM for hPD-L1 and mPD-L1, respectively). In the HepG2/Jurkat T co-culture cell model, Z13 decreased the viability rate of HepG2 cells in a concentration-dependent manner. In addition, Z13 showed significant in vivo antitumor efficacy (TGI = 52.6 % at 40 mg/kg) without obvious toxicity in the B16-F10 melanoma model. Furthermore, flow cytometry analysis demonstrated that Z13 inhibited tumor growth in vivo by activating the tumor immune microenvironment. These findings indicate that Z13 is a promising PD-1/PD-L1 inhibitor deserving further investigation.

Salicylic acid doped silica nanoparticles as a fluorescent nanosensor for the detection of Fe3+ in aqueous solution.

A novel organic-inorganic hybrid nanosensor (SASP) was prepared by a one-step sol-gel method and characterized by scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis, N2 adsorption-desorption, fluorescence spectroscopy, etc. The nanosensor showed almost 3-fold fluorescence emission quenching upon excitation with a 293 nm wavelength in the presence of 20 µM Fe3+ ions. The presence of 18 other metal ions had no observable effect on the sensitivity and selectivity of the nanosensor. A fluorescence analysis method based on the SASP for the selective detection of Fe3+ was established under optimal conditions. The results showed that there was a linear relationship between the log luminescence value and the concentration of Fe3+ over the range of 2.0 × 10-7-9.0 × 10-5 mol L-1 with a detection limit (3σ) of 2.5 × 10-8 mol L-1. Furthermore, the proposed method was successfully applied for the determination of trace Fe3+ in fetal bovine serum without the interference of other molecules and ions. Good recovery (96.5-104.5%) and a relative standard deviation of less than 8.6% were obtained from serum samples spiked with four levels of Fe3+. Additionally, the nanosensor showed a good reversibility; the fluorescence could be switched "off" and "on" in two ways, by adjusting the pH of the solution and adding metal chelating agent EDTA.

Development of hydroxypropyl cellulose and graphene oxide modified molecularly imprinted polymers for separation and enrichment of podophyllotoxin.

A novel hydroxylpropyl cellulose (HPC) modified graphene oxide (GO)-based molecularly imprinted polymers (HPC-GO-MIP) have been developed as a solid phase extraction (SPE) material for the selective separation and extraction of podophyllotoxin. In this strategy, the cellulose with rich hydroxyl groups was introduced to form bi-functional monomers with methacrylic acid to provide more recognition sites for the improving of extraction efficiency, then GO was added as a two-dimensional substrate for MIP to improve the material morphology and surface area. The extraction performances of obtained HPC-GO-MIP material were tested, and the results prove its high efficiency and selectivity for podophyllotoxin extraction. The saturated adsorption capacity reached 23.1 µg/mg, and high enrichment efiiciency of 463.8 folds was realized under the premise of ensuring the recovery rate. The selective imprinting factor was much higher than those of kaempferol and quercetin, which were the main compounds in podophyllum fruit. Under the optimized SPE conditions, the HPC-GO-MIP based SPE-HPLC method showed the detection limit of 14.2 ng/mL for podophyllotoxin assay. When applied to podophyllum fruit samples, the material showed excellent ability of selective separation and enrichment of podophyllotoxin, and the relative standard deviations (RSD) of intra and inter batches were less than 8.1 % and 5.7 % in real samples detection. The HPC-GO-MIP SPE method broadened the application for high multiple extraction in trace analyte samples and provided a valuable solution to improve the selective separation and detection.

A quantum dot aptamer fluorescent sensor based on magnetic graphene oxide for the detection of zearalenone.

As an estrogenic mycotoxin found in a wide range of agricultural crops, the toxicity of zearalenone (ZEN) poses a serious risk to human health. Accordingly, to achieve rapid detection of zearalenone in complex samples, an aptamer fluorescence sensor based on magnetic graphene oxide was developed. Compared with traditional methods, this technique has the virtues of simple operation, low cost, and reliable performance. Magnetic graphene oxide (MGO) was synthesized as a fluorescent bursting agent, using a chemical precipitation approach by depositing Fe3O4 on the surface of graphene oxide. As a fluorescent probe, an aptamer coupling with CdTe quantum dots and zearalenone was used. Following the specific binding of zearalenone and the aptamer, the affinity interaction between the fluorescent probe and MGO was weakened, resulting in the recovery of fluorescence and making the qualitative and quantitative analysis of zearalenone available via fluorescence intensity determination. The results indicated that the method's linear range was 5-120 µg L-1 and its detection limit was 2.9 µg L-1. In addition, the recoveries varied from 76.4 to 118.8% for crop samples, with relative standard deviations (RSDs) between 0.8 and 9.5%, which suggests an effective method for the separation and detection of mycotoxins in actual environmental samples.

Fabrication of magnetic molecularly imprinted polymers based on aptamers and ß-cyclodextrin for synergistic recognition and separation of tetracycline.

Tetracycline is extensively used as an antibiotic in animal husbandry, and there arose an increase in antibiotic resistance genes in the environment, posing a threat to human health. Motivated by this, a magnetic molecularly imprinted material based on synergistic recognition (1 + 1>2) was constructed and coupled with high-performance liquid chromatography to detect ultra-trace tetracycline in complicated samples. In this case, the molecularly imprinted polymers were synthesized via a "one-pot" method and acted as recognition elements on the surface of silica-coated ferroferric oxide particles. Aptamers and ß-cyclodextrin, as functional monomers, had a synergistic effect on the recognition of tetracycline and the synergistic recognition factor was 1.7. Meanwhile, the material exhibited high selectivity to tetracycline with an imprinting factor of 7.6. In addition, compared to being modified on the surface, the stability of the aptamers was effectively improved by cross-linking in the molecularly imprinted polymer framework. Relevant experimental conditions, such as buffers, concentration of magnesium ions and adsorption time, were optimized. As a result, the method showed a limit of detection of 1.0 µg L-1 and the linearity range of 0.005-0.5 mg L-1, as well as certain reproducibility and stability. Furthermore, when applied for the analysis of animal feed samples, a significant reduction of matrix interference was observed with satisfactory recoveries (85.0-111.5%), which emphasized the good accuracy and practicability of the established method. For these advantages, the proposed method represents a versatile and powerful tool for the separation and detection of small molecule compounds in complicated real samples.

Biodegradability and Cytocompatibility of 3D-Printed Mg-Ti Interpenetrating Phase Composites.

Orthopedic hybrid implants combining both titanium (Ti) and magnesium (Mg) have gained wide attraction nowadays. However, it still remains a huge challenge in the fabrication of Mg-Ti composites because of the different temperatures of Ti melting point and pure Mg volatilization point. In this study, we successfully fabricated a new Mg-Ti composite with bi-continuous interpenetrating phase architecture by infiltrating Mg melt into Ti scaffolds, which were prepared by 3D printing and subsequent acid treatment. We attempted to understand the 7-day degradation process of the Mg-Ti composite and examine the different Mg2+ concentration composite impacts on the MC3T3-E1 cells, including toxicity, morphology, apoptosis, and osteogenic activity. CCK-8 results indicated cytotoxicity and absence of the Mg-Ti composite during 7-day degradation. Moreover, the composite significantly improved the morphology, reduced the apoptosis rate, and enhanced the osteogenic activity of MC3T3-E1 cells. The favorable impacts might be attributed to the appropriate Mg2+ concentration of the extracts. The results on varying Mg2+ concentration tests indicated that Mg2+ showed no cell adverse effect under 10-mM concentration. The 8-mM group exhibited the best cell morphology, minimum apoptosis rate, and maximum osteogenic activity. This work may open a new perspective on the development and biomedical applications for Mg-Ti composites.

Experimental and DFT investigation on N-functionalized biochars for enhanced removal of Cr(VI).

In this study, N-functionalized biochars with varied structural characteristics were designed by loading poplar leaf with different amounts of urea at 1:1 and 1:3 ratios through pyrolysis method. The addition of urea significantly increased the N content of biochar and facilitated the formation of amine (-NH-, -NH2), imine (-HCNH), benzimidazole (-C7H5N2), imidazole (-C3H3N2), and pyrimidine (-C4H3N2) groups due to substitution reaction and Maillard reaction. The effect of pH on Cr(VI) removal suggested that decrease in solution pH favored the formation of electrostatic attraction between the protonated functional groups and HCrO4-. And, experimental and density functional theory study were used to probe adsorption behaviors and adsorption mechanism which N-functionalized biochars interacted with Cr(VI). The protonation energy calculations indicated that N atoms in newly formed N-containing groups were better proton acceptors. Adsorption kinetics and isotherm experiments exhibited that N-functionalized biochars had greater removal rate and removal capacity for Cr(VI). The removal rate of Cr(VI) on N-functionalized biochar was 10.5-15.5 times that of untreated biochar. Meanwhile, N-functionalized biochar of NB3 with the largest number of adsorption sites for -C7H5N2, -NH2, -OH, -C3H3N2, and phthalic acid (-C8H5O4) exhibited the supreme adsorption capacity for Cr(VI) through H bonds and the highest adsorption energy was -5.01 kcal/mol. These mechanistic findings on the protonation and adsorption capacity are useful for better understanding the functions of N-functionalized biochars, thereby providing a guide for their use in various environmental applications.

A dispersive solid phase extraction adsorbent based on aptamer modified chitosan nanofibers for zearalenone separation in corn, wheat, and beer samples.

Highly selective separation of trace bio-toxins in food samples has long been a hot topic pursued by analytical chemists. In this paper, chitosan nanofibers prepared by freeze-drying were modified with aptamers for dispersive solid phase extraction (dSPE) of trace zearalenone. The morphology of achieved chitosan nanofibers was found to be uniform and continuous, and the length was at the micron level with about a 400 nm diameter. The immobilization capacity of the aptamer was as high as 10.1 µg on 5 mg chitosan nanofibers with good stability and repeatability, owing to the high specific surface area of nanofibers. The aptamer modified chitosan nanofibers (Apt-CNFs) showed specific selectivity to zearalenone with a selectivity coefficient of 2.65 compared to the scrambled oligonucleotide functionalized CNFs, and the selectivity factors over other analogs and reference compounds were from 1.57 to 50.0. After the optimization of extraction conditions, the Apt-CNF based dSPE was coupled with high-performance liquid chromatography for zearalenone monitoring, and a good linear range of 0.06-10.0 µg L-1 was achieved with a detection limit of 18.0 ng L-1. The spiking recoveries of 101-108%, 100-110%, and 98.3-101% were achieved for trace zearalenone in corn, wheat, and beer samples, respectively. The residual zearalenone was detected in corn and wheat with a content of 0.365 and 0.0775 µg g-1, respectively.

Key factors and microscopic mechanisms controlling adsorption of cadmium by surface oxidized and aminated biochars.

Modified biochar has great potential for adsorbing cadmium (Cd) in the aquatic environment, but the micro-immobilization mechanisms, driven by surface modifications, remain unclear. There has been no attempt to determine the key adsorption factors by integrating the numerous physiochemical indicators. In this study, surface oxidized biochar (OPBC) and surface aminated biochar (APBC) were prepared from porous biochar (PBC), and the Cd adsorption mechanisms by the modified biochars at the molecular and electronic scales were investigated. The adsorption capacity of APBC and OPBC for Cd was 23.54 and 19.04 mg g-1, respectively, which was about three times higher than that of PBC. Macroscopically, physicochemical adsorption and intraparticle diffusion dominated the Cd adsorption, and surface properties, such as functional groups, were identified as key factors controlling adsorption. Microscopically, the adsorption of Cd mainly occurred in regions rich in π electrons, lone pair electrons and electron donor groups. The interaction between carboxyl and Cd dominated the adsorption performance of OPBC, while the Cd2+-π interaction was weakened by increasing the π electron electrostatic potential of aromatic rings. The lone pair electrons of the amino groups dominated the complexation of APBC with Cd, and the π electron electrostatic potential was almost unaffected.

Determination of Pesticides by Gas Chromatography Combined with Mass Spectrometry Using Femtosecond Lasers Emitting at 267, 400, and 800 nm as the Ionization Source.

A standard sample mixture containing 51 pesticides was separated by gas chromatography (GC), and the constituents were identified by mass spectrometry (MS) using femtosecond lasers emitting at 267, 400, and 800 nm as the ionization source. A two-dimensional display of the GC/MS was successfully used for the determination of these compounds. A molecular ion was observed for 38 of the compounds at 267 nm and for 30 of the compounds at 800 nm, in contrast to 27 among 50 compounds when electron ionization was used. These results suggest that the ultraviolet laser is superior to the near-infrared laser for molecular weight determinations and for a more reliable analysis of these compounds. In order to study the conditions for optimal ionization, the experimental data were examined using the spectral properties (i.e., the excitation and ionization energies and absorption spectra for the neutral and ionized species) obtained by quantum chemical calculations. A few molecules remained unexplained by the currently reported rules, requiring additional rules for developing a full understanding of the femtosecond ionization process. The pesticides in the homogenized matrix obtained from kabosu ( citrus sphaerocarpa) were measured using lasers emitting at 267 and 800 nm. The pesticides were clearly separated and measured on the two-dimensional display, especially for the data measured at 267 nm, suggesting that this technique would have potential for use in the practical trace analysis of the pesticides in the environment.

Characterization and 2D structural model of corn straw and poplar leaf biochars.

The integrated experimental methods were used to analyze the physicochemical properties and structural characteristics and to build the 2D structural model of two kinds of biochars. Corn straw and poplar leaf biochars were gained by pyrolysing the raw materials slowly in a furnace at 300, 500, and 700 °C under oxygen-deficient conditions. Scanning electron microscope was applied to observe the surface morphology of the biochars. High temperatures destroyed the pore structures of the biochars, forming a particle mixture of varying sizes. The ash content, yield, pH, and surface area were also observed to describe the biochars' properties. The yield decreases as the pyrolysis temperature increases. The biochars are neutral to alkaline. The biggest surface area is 251.11 m2/g for 700 °C corn straw biochar. Elemental analysis, infrared microspectroscopy, solid-state C-13 NMR spectroscopy, and pyrolysis gas chromatography-mass spectrometry (Py-GC-MS) were also used to study the structural characteristics and build the 2D structural models of biochars. The C content in the corn straw and poplar leaf biochars increases with the increase of the pyrolysis temperature. A higher pyrolysis temperature makes the aryl carbon increase, and C=O, OH, and aliphatic hydrocarbon content decrease in the IR spectra. Solid-state C-13 NMR spectra show that a higher pyrolysis temperature makes the alkyl carbon and alkoxy carbon decrease and the aryl carbon increase. The results of IR microspectra and solid-state C-13 NMR spectra reveal that some noticeable differences exist in these two kinds of biochars and in the same type of biochar but under different pyrolysis temperatures. The conceptual elemental compositions of 500 °C corn straw and poplar leaf biochars are C61H33NO13 and C59H41N3O12, respectively. Significant differences exist in the SEM images, physicochemical properties, and structural characteristics of corn straw and poplar leaf biochars.

Adsorption Mechanisms of Dodecylbenzene Sulfonic Acid by Corn Straw and Poplar Leaf Biochars.

Biochar is an eco-friendly, renewable, and cost-effective material that can be used as an adsorbent for the remediation of contaminated environments. In this paper, two types of biochar were prepared through corn straw and poplar leaf pyrolysis at 300 °C and 700 °C (C300, C700, P300, P700). Brunaer-Emmett-Teller N2 surface area, scanning electron microscope, elemental analysis, and infrared spectra were used to characterize their structures. These biochars were then used as adsorbents for the adsorption of dodecylbenzene sulfonic acid (DBSA). The microscopic adsorption mechanisms were studied by using infrared spectra, 13C-nuclear magnetic resonance spectra, and electron spin resonance spectra. The surface area and pore volume of C700 (375.89 m²/g and 0.2302 cm³/g) were the highest among all samples. Elemental analysis results showed that corn straw biochars had a higher aromaticity and carbon to nitrogen (C/N) ratio than the poplar leaf biochars. High temperature caused the increase of carbon content and the decrease of oxygen content, which also gave the biochars a higher adsorption rate. Pseudo-second order kinetic provided a better fit with the experimental data. Adsorption isotherm experiments showed that the adsorption isotherm of C300 fit the linear model. For other biochars, the adsorption isotherms fitted Langmuir model. Biochars with high temperatures exhibited enhanced adsorption capacity compared with ones at low temperatures. The qmax values of biochars to DBSA followed the order of P700 > C700 > P300. The adsorption mechanisms were complex, including partition, anion exchange, the formation of H bonds, covalent bonds, and charge transfer. The adsorption by covalent bonding might be the key mechanism determining the adsorption capacity of P700.

Bio-inspired energy-harvesting mechanisms and patterns of dynamic soaring.

Albatrosses can make use of the dynamic soaring technique extracting energy from the wind field to achieve large-scale movement without a flap, which stimulates interest in effortless flight with small unmanned aerial vehicles (UAVs). However, mechanisms of energy harvesting in terms of the energy transfer from the wind to the flyer (albatross or UAV) are still indeterminate and controversial when using different reference frames in previous studies. In this paper, the classical four-phase Rayleigh cycle, includes sequentially upwind climb, downwind turn, downwind dive and upwind turn, is introduced in analyses of energy gain with the albatross's equation of motions and the simulated trajectory in dynamic soaring. Analytical and numerical results indicate that the energy gain in the air-relative frame mostly originates from large wind gradients at lower part of the climb and dive, while the energy gain in the inertial frame comes from the lift vector inclined to the wind speed direction during the climb, dive and downwind turn at higher altitude. These two energy-gain mechanisms are not equivalent in terms of energy sources and reference frames but have to be simultaneously satisfied in terms of the energy-neutral dynamic soaring cycle. For each reference frame, energy-loss phases are necessary to connect energy-gain ones. Based on these four essential phases in dynamic soaring and the albatrosses' flight trajectory, different dynamic soaring patterns are schematically depicted and corresponding optimal trajectories are computed. The optimal dynamic soaring trajectories are classified into two closed patterns including 'O' shape and '8' shape, and four travelling patterns including 'Ω' shape, 'α' shape, 'C' shape and 'S' shape. The correlation among these patterns are analysed and discussed. The completeness of the classification for different patterns is confirmed by listing and summarising dynamic soaring trajectories shown in studies over the past decades.

Determination of Hexachlorocyclohexane by Gas Chromatography Combined with Femtosecond Laser Ionization Mass Spectrometry.

Structural isomers and enantiomers of hexachlorocyclohexane (HCH) were separated using a chiral column by gas chromatography and quantitatively determined by multiphoton ionization mass spectrometry using an ultraviolet femtosecond laser (200 and 267 nm) as the ionization source. The order of elution of the enantiomers (i.e., (+)-α-HCH and (-)-α-HCH) was predicted from stabilization energies calculated for the complexes using permethylated γ-cyclodextrin as the stationary phase of the column, and the results were compared with the experimental data. The molecular ions observed for HCH were weak, even though they can be ionized through a process of resonance enhanced two-photon ionization at 200 nm. This unfavorable result can be attributed to the dissociation of the molecular ion, as predicted from quantum chemical calculations. Graphical Abstract ᅟ.

Distribution and content of lipid droplets and mitochondria in pig parthenogenetically activated embryos after delipation.

The present study examines the effect of delipation on developmental competence and the distribution pattern of lipid droplets (LDs) and mitochondria in parthenogenetically activated (PA) pig embryos. Mature oocytes were delipated by centrifugation after partial digestion of the zonae pellucidae, subjected to parthenogenetic activation after total removal of zonae pellucidae by pronase, and then cultured in vitro up to the blastocyst stage. The contents and distributions of LDs and mitochondria in the oocytes and/or embryos were observed by staining with Oil Red O and MitoTracker Red CMXRos, respectively. The LD and mitochondrial contents were significantly reduced by the delipation process, and only smaller LDs remained in the delipated oocytes and/or embryos. Their content remained constant from the metaphase II oocyte to the blastocyst stage, but they became gradually smaller as the oocytes and/or embryos developed. The distribution pattern of the LDs in the delipated embryos changed over time and in a manner different to that seen in the controls. In the early developmental stages (1- to 4-cell stages), they were distributed peripherally and formed a ring around the nucleus. However, by the blastocyst stage, a homogeneous distribution of LDs was observed in both the inner cell mass and trophectoderm. The distribution pattern of mitochondria also changed with the development of the delipated PA embryos and again, in ways different to those seen in the controls. In the early 1- to 4-cell stages, a peripheral distribution of mitochondrial foci was observed in each blastomere. However, in blastocysts, the mitochondria were homogeneously distributed throughout the inner cell mass and trophectoderm. Although the cleavage rate at the 2- and 4-cell stages of the PA embryos was not affected by delipation (95.83 ± 2.25% vs. 97.44 ± 0.67%; 79.17 ± 4.47% vs. 84.62 ± 1.19%), it was reduced significantly in the blastocyst compared with the controls (21.67 ± 3.78% vs. 49.36 ± 1.77%). The distribution pattern of the LDs in oocytes and/or embryos at different developmental stages, and that of the mitochondria in metaphase II oocytes, was affected by delipation. The developmental competence of porcine PA embryos would appear to be affected by delipation.