Fabricating Ultrathin Imprinting Layer for Fast Capture of Valsartan via a Metal Affinity-Oriented Surface Imprinting Method.

Rapid separation and enrichment of targets in biological matrixes are of significant interest in multiple life sciences disciplines. Molecularly imprinted polymers (MIPs) have vital applications in extraction and sample cleanup owing to their excellent specificity and selectivity. However, the low mass transfer rate, caused by the heterogeneity of imprinted cavities in polymer networks and strong driving forces, significantly limits its application in high-throughput analysis. Herein, one novel metal affinity-oriented surface imprinting method was proposed to fabricate an MIP with an ultrathin imprinting layer. MIPs were prepared by immobilized template molecules on magnetic nanoparticles (NPs) with metal ions as bridges via coordination, and then polymerization was done. Under the optimized conditions, the thickness of the imprinting layer was merely 1 nm, and the adsorption toward VAL well matched the Langmuir model. Moreover, it took just 5 min to achieve adsorption equilibrium significantly faster than other reported MIPs toward VAL. Adsorption capacity still can reach 25.3 mg/g ascribed to the high imprinting efficiency of the method (the imprinting factor was as high as 5). All evidence proved that recognition sites were all external cavities and were evenly distributed on the surface of the NPs. The obtained MIP NPs exhibited excellent selectivity and specificity toward VAL, with good dispersibility and stability. Coupled with high-performance liquid chromatography, it was successfully used as a dispersed solid phase extraction material to determine VAL in serum. Average recoveries are over 90.0% with relative standard deviations less than 2.14% at three spiked levels (n = 3). All evidence testified that the MIPs fabricated with the proposed method showed a fast trans mass rate and a large rebinding capacity. The method can potentially use high-throughput separation and enrichment of target molecules in batch samples to meet practical applications.

Bilberry Anthocyanins (Vaccinium myrtillus L.) Induced Apoptosis of B16-F10 Cells and Diminished the Effect of Dacarbazine.

Anthocyanins have been reported to have potential as dietary or pharmaceutical supplements in the application of cancer prevention and adjunctive treatment. However, there are few studies on the effect of anthocyanins on melanoma, which have only been performed in cell lines. The objective of this work was to investigate the anticancer effects and mechanisms of bilberry anthocyanin extract (BAE) on melanoma In Vitro and In Vivo. Moreover, a primary study was done to investigate how BAE influenced C57BL/6 mice bearing subcutaneous B16-F10 tumors treated with dacarbazine (DTIC). BAE-induced apoptosis in B16-F10 cells was associated with activation of the mitochondrial pathway induced by increased reactive oxygen species. More, In Vivo anticancer activity studies indicated that BAE attenuated melanoma growth, as identified by hematoxylin-eosin staining, Ki-67, and TUNEL assays. Further western blot results revealed higher phospho-Akt expression with the combination of BAE and DTIC, indicating no suppression of the PI3K/AKT signaling pathway. In summary, this study demonstrated the anti-melanoma activity of BAE and investigated its mechanism. Notably, it should be careful to use products enriching BAE for those melanoma patients treated with DTIC.

Facile Way to Prepare a Porous Molecular Imprinting Lock for Specifically Recognizing Oxytetracyclin Based on Coordination.

Molecularly imprinted polymers (MIPs) are a kind of synthetic receptor-like materials. They have drawn more and more attention in the past decades. In this work, a facile method was developed to prepare porous magnetic MIPs utilizing metal coordination. The preparation is simply done using conventional oil-in-water emulsifier-free emulsion technology by mixing poly(styrene-co-itaconic acid), oxytetracyclin (OTC), Cu(II), and Fe3O4 magnetic fluid in one pot with a reaction time of 3 h. The product shows high specificity and selectivity toward OTC, as well as an excellent saturation adsorption capacity (62.567 mg/g). Emphasizing that the imprinting factor is 29, which is the highest one among the reported MIPs to the best of our knowledge. Combined with high-performance liquid chromatography, it was used successfully to determine OTC in pork liver, one of the most complex bio-samples. Recoveries are higher than 91.0% with relative standard deviations less than 4.5% at three spiked levels (n = 3). All evidence testifies that the MIPs based on metal coordination show excellent recognition selectivity and specificity, as well as large rebinding capacity. The strategy holds promise as a reliable, extensible, and versatile way for preparing a metal ion-mediated molecular-imprinting polymer.

A glassy carbon electrode modified with a monolayer of zirconium(IV) phosphonate for sensing of methyl-parathion by square wave voltammetry.

A glassy carbon electrode (GCE) was consecutively modified with amino groups and phosphate groups, and then loaded with Zr(IV) ions. Fourier transform infrared spectrophotometry, field-emission scanning electron microscopy, energy dispersive X-ray spectroscopy and cyclic voltammetry were used to characterize the morphologies and electrochemical properties. The sensor was used to detect p-nitrophenyl-substituted organophosphorus pesticides, with methyl-parathion (MP) as the model analyte. Under optimized conditions, the oxidation current of square wave voltammetry (typically measured at around -0.28 V vs. saturated calomel electrode) increases linearly in the 1.0 to 100 ng mL-1 MP concentration range, and the detection limit is 0.25 ng mL-1 (at a signal to noise ratio of 3). Average recoveries from (spiked) real water samples are 99.9-102.2%, with relative standard deviations of 0.3-2.6% (n = 3) at three levels. The reliability and accuracy of the method was validated by HPLC. Graphical abstract Zr(IV) modified GCE is prepared via three steps. The electrode shows high specificity and selectivity towards methyl-parathion. And the linear range is 1.0 - 100.0 ng mL-1 with the detection limit as low as 0.25 ng mL-1 with SWV.

Molecularly imprinted electrochemical sensor based on metal-covalent organic framework for specifically recognizing norfloxacin from unpretreated milk.

Here, a molecularly imprinted electrochemical sensor (MIECS) was designed and fabricated for specifically monitoring norfloxacin (NFX), an entirely synthetic antibiotic. In which, Cu2+ dopped covalent organic framework (COF) was used to connect NFX imprinting layer and glassy carbon electrode through covalence. Under optimized conditions, the linear range is as wide as 5 orders of magnitude, and the detection limit is 5.94 × 10-3 µM (estimated based on S/N = 3). Average recoveries are among 92.4%-99.0% with relative standard deviations ≤ 4.05% (n = 3) in (spiked) whole, low-fat, and skimmed milk, validated by independent HPLC assays. The excellent performance can be ascribed to the significant recognition and enriching ability of the imprinting layer, improved conductivity of Cu2+ dopped covalent organic framework, and high stability of covalence between layers. We hope the work will act as a model of MIECSs for rapidly and selectively detecting trace drug residue in complex real samples.

Efficient separation of aristolochic acid I from Caulis aristolochiae manshuriensis (Guan-mu-tong) with copper mediated magnetic molecularly imprinted polymer.

Screening bioactive compounds from natural products is one of the most effective ways for new drug research and development. However, obtaining a single extract component on a large scale and with high purity from a complex matrix is still an arduous and challenging task. Herein, one metal mediated magnetic molecularly imprinted polymer (mMIP) was rationally designed and prepared for specifically capturing Aristolochic acid I (AAI). The preparation was done with copper(II) as binding pivot, (3-aminopropyl) triethoxysilane as functional monomer, and Fe3O4 as core, by a one-step sol-gel method. Under the optimized conditions, the apparent maximum binding amount of copper mediated mMIP (Cu-mMIP) reaches as high as 349.72 mg g-1, the highest among the reported AAI-MIPs. Moreover, the nanoparticles exhibit excellent specificity and selectivity, good reproducibility and stability, high superparamagnetism (60.32 emu g-1), and high imprinting efficiency (an imprinting factor of 7). By simulating an industrial-scale separation, 16.56 mg AAI (purity of 95.11%) is obtained after six cycles with 100 mg nanoparticles from 20 g Caulis aristolochiae manshuriensis (Guan-mu-tong). Notably, this takes only 3 hours and consumes 50 mL of methanol. The study provides a potent tool for the green, fast, and specific extraction of high-purity ingredients from natural plants in the manufacturing industry and conventional analysis in the lab.

Molecular Charge and Antibacterial Performance Relationships of Aggregation-Induced Emission Photosensitizers.

Bacterial infections remain a major cause of morbidity worldwide due to drug resistance of pathogenic bacteria. Photodynamic therapy (PDT) has emerged as a promising approach to overcome this drug resistance. However, existing photosensitizers (PSs) are broad-spectrum antibacterial agents that dysregulate the microflora balance resulting in undesirable side effects. Herein, we synthesized a series of aggregation-induced emission (AIE)-active PSs with a lipophilic cationic AIE core with varying charges, named TBTCP and its derivatives. The association of the difference in their molecular charge with the antibacterial effects was systemically investigated. Among the derivatives presented, TBTCP-SF with the electronegative sulfonate group nulled its ability to bind to and ablate Gram-positive (G+) or Gram-negative (G-) bacteria. TBTCP-QY modified by electropositive quaternary ammonium facilitated binding and augmented the photodynamic antibacterial activity for both G+ and G- bacteria. TBTCP-PEG with hydrophilic neutral ligands selectively bound and inactivated G+ bacteria. Under white-light illumination, TBTCP-PEG ablated 99.9% methicillin-resistant Staphylococcus aureus (MRSA) and promoted wound healing in MRSA-infected mice, eliminating MRSA infection both in vitro and in vivo. Our work provides unprecedented insight into the utility of AIE-active PSs for highly targeted and efficient photodynamic ablation of either G+ or G- bacteria that can be translated to next-generation antibacterial materials.

A human VEGF magnetic molecularly imprinted polymer for drug-free anti-angiogenesis and photothermal therapy of tumors.

A magnetic molecularly imprinted polymer was developed with an epitope peptide of human VEGF as a template via an epitope blotting technique. As a drug-free agent, the nanoparticles can significantly suppress the proliferation of tumor cells by integrating anti-angiogenesis and photothermotherapy. This work provides a successful example of the design of multimodal antineoplastic drugs.

Copper mediated molecularly imprinted polymers for fast recognizing tylosin.

As one kind of artificial antibody, molecularly imprinted polymers (MIPs) has been widely used to separate/enrich target molecules from samples with the complex matrix prior to instrumental analysis. In this work, one novel copper mediated magnetic MIPs was developed towards anti-bacteria macrolide antibiotic tylosin (TYL). The obtained microspheres had a lot of convexities distributed evenly on the surface. And it exhibited high hydrophilicity and superparamagnetic ability, as well as excellent selectivity and specificity. Notably, it only took 5 min to reach the absorption equilibrium for TYL, which made it highly feasible for high-throughput analysis. Furthermore, the microspheres was applied as the magnetic dispersed solid phase extraction material to enrich trace TYL residue in (spiked) human urine. At three spiking levels, mean recoveries are in the range of 76.42-93.72% with relative standard deviations of 2.75-8.24% (n = 3) with HPLC/UV-Vis. The work provided one promising reference to prepare novel solid phase extraction materials for enriching/separating trace component in complex matrix.

[Preparation of molecularly imprinted polymers based on covalent organic frameworks and their application to selective recognition of trace norfloxacin in milk].

Norfloxacin (NFX) is an antibiotic that is widely used in animal husbandry. However, the presence of NFX even in trace amounts in animal-derived food may harm human health. Therefore, it is of practical significance to establish a method for monitoring NFX residues in food. Molecularly imprinted polymers (MIPs) imitate interactions established by natural receptors to selectively retain a target molecule, like antibodies or antigens do. MIPs have been widely employed in the selective recognition of specific target molecules from complex samples. Covalent organic frameworks (COFs) are a new type of organic polymer with uniform and ordered crystal structures. COFs form crystal structures by constructing organic units for ordered assembly through reversible chemical reactions. Their porous structure, regular morphology, and easy modification make COFs promising for use as excellent adsorbent carriers. Owing to these advantages of COFs, researchers have attempted to coat one MIP layer on COFs; however, the preparation methods are time-consuming and laborious, and the conditions are harsh. Hence, this study proposes a simple and rapid method for the preparation of novel MIPs with COFs as the support (DP-COF@MIPs) for the selective recognition of NFX. First, a Schiff-base COF (DP-COF) was rapidly synthesized using 3,3'-diaminobenzidine and p-phthalaldehyde with a metal trifluorate as a catalyst at room temperature. Subsequently, a two-step sequence was adopted as the synthesis strategy using NFX as the template, methacrylic acid as a functional monomer, and ethylene glycol dimethacrylate as a crosslinking agent. The entire synthesis was completed within 5 h under mild conditions. The material was then characterized by multiple analytical methods, including field-emission scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, and the Brunauer-Emmett-Teller (BET) method (to determine the specific surface area). The experimental results confirmed the successful preparation of DP-COF@MIPs. The DP-COF@MIPs presented a rough and porous surface, with a pore size of approximately 17.79 nm (mesoporous structure). The properties of the material were evaluated by adsorption and regeneration experiments. The kinetic adsorption experiment showed that the DP-COF@MIPs reached adsorption equilibrium in 90 min. Two straight lines were fitted using the pseudo-second-order kinetics model based on the experimental results for thermodynamic adsorption, indicating that the adsorption process was primarily dominated by chemical adsorption. The maximum apparent adsorption capacity was 41.57 mg/g. In the selective and competitive binding test, five drugs, namely ciprofloxacin, dimetridazole, oxytetracycline, sulfadiazine, and chloramphenicol, were selected as the interferents. The experimental results showed that the DP-COF@MIPs possessed good stereoselectivity and competitive recognition ability. The regeneration of DP-COF@MIPs was evaluated by multiple cycles of adsorption-desorption experiments. The loss in the adsorption capacity of the particles was only approximately 4.7% after seven adsorption-desorption cycles. These results from the regeneration experiments show that the DP-COF@MIPs had high stability and reusability in the selective adsorption and separation of NFX. In addition, the employed method could accurately identify trace NFX in milk samples. The average recoveries were in the range of 88.8%-92.9% at three spiked levels (0.03, 0.10, and 0.30 mg/L) with relative standard deviations (RSDs) in the range of 0.6-1.7% (n=3). Notably, the method could successfully determine NFX at contents as low as 0.0020 mg/L in the milk sample with an average recovery of 77.6% and RSD of 6.4% (n=3). This concentration is one-fiftieth of the maximum residue level stipulated by the European Union (EU), and even lower than the limit of detection (LOD) of conventional high performance liquid chromatography (HPLC) methods. The above results confirm that DP-COF@MIPs can be used to determine trace NFX in actual complex samples using HPLC equipment, even when coupled to conventional UV-Vis detectors. This study offers a facile and general method for the preparation of MIPs based on COFs with selective recognition ability.

Designing and preparing metal mediated magnetic imprinted polymer for recognition of tetracycline.

Recently, metal mediated molecularly imprinted polymers (MMIPs) raise extensive attention due to their special adsorption/desorption mechanism. And the metal ion plays a key role both for MMIPs preparation and molecular recognition. But it is still a big question to select one suitable metal ion. To overcome above problem, one computational approach was proposed by calculating interaction energy (ΔE) of ternary complexes (monomer-M2+-template) with GAMESS software at the PBE0 level. Finally Cu(II) was screened from four doubly charged metal ions with different radii. Then Cu mediated MIP was prepared via surface imprinting technique with Fe3O4 as the core in a water system. Subsequently, the adsorption behavior of the MMIP immobilized Cu(II) was investigated in detail. The results indicated the microspheres own excellent specificity and selectivity towards tetracycline (TC), as well as long-term stability and good reproducibility. The apparent maximum binding capacity and the imprinting factor were as high as 163.5 mg g- 1 and 19, respectively. Coupled with HPLC-PDA, the microspheres was successfully used as a dispersed solid phase extraction material to extract trace TC in chicken liver. At three spiking levels, mean recoveries ranged from 89.3% to 97.1% with relative standard deviations less than 4.0% (n = 3). Limits of detection and quantitation were 0.0073 mg kg- 1 and 0.024 mg kg- 1, respectively, which can meet the strict requirement of the regulations stipulated by EU. Moreover, preliminary studies showed the radius of the metal ion and ΔE of the complexes all affect the adsorption capacity, specificity and mass transfer rate of the MMIP.

Synergistically Enhancing Tumor Chemotherapy Using an Aggregation-Induced Emission Photosensitizer on Covalently Conjugated Molecularly Imprinted Polymer Nanoparticles.

Due to the polygenic and heterogeneous nature of the tumorigenesis process, traditional chemotherapy is far from desirable. Fabricating multifunctional nanoplatforms integrating photodynamic effect can synergistically enhance chemotherapy because they can make the cancer cells much sensitive to chemotherapeutics. However, how to assemble different units in nanoplatforms and minimize side effects caused by chemodrugs and photosensitizers (PSs) still needs to be explored. Herein, a nanoplatform CPP/PS-MIP@DOX is developed using a simultaneously covalently conjugated new aggregation-induced emission (AIE) PS and a cell-penetrating peptide (CPP) on the surface of silica-based molecularly imprinted polymer (MIP) nanoparticles, prepared with doxorubicin (DOX) as the template in the water system via a sol-gel technique. CPP/PS-MIP@DOX has good biocompatibility, high DOX-loading ability, promoted cellular uptake, and sustained and pH-sensitive drug release capability. Furthermore, it can efficiently penetrate into tumor tissue, accurately home to, and accumulate at the tumor site. As a result, a better efficacy with lower cytotoxicity is achieved with a smaller dosage of DOX by utilizing either the photodynamic effect or unique characteristics of the MIP. It is the first nanoplatform fabricated by chemically conjugating AIE PSs directly on the surface of the scaffold via the surface-decorated strategy and successfully applied in cancer therapy. This work provides an effective strategy by constructing AIE PS-based cancer nanomedicines with MIPs as scaffolds.

Biomonitoring human urinary levels of 26 metal elements in multi-race coexistence region of Xinjiang, China.

The human biological monitoring of metals (metalloids) is of importance, which concentrations could indicate a wide range of health related information. Reference values (RVs) of metals (metalloids) in specific matix of populations are critically required when performing a statistical evaluation accurately. Recent studies show that RVs of metals (metalloids) are influenced by many multiple factors, including lifestyle, diet, dissimilar environment, location, as well as race. However, it is virtually absent across many nations/regions, especially multi-race coexistence regions. The aim of the work was to establish RVs of metals (metalloids) in urine of adult in Xinjiang, China. Totally 178 urine samples from healthy volunteers were collected and analyzed by inductively coupled plasma mass spectrometer or optical emission spectrometer (ICP-MS/OES). RVs of 26 metals (metalloids) in urine for adult of Xinjiang, China were then established. Effects of race, gender and age on RVs were investigated using partial least squares discriminant analysis and student T-test. The result demonstrated that above factors showed mild influence for the establishment of RVs. The RVs derived from total 178 urine adult samples (Han and Uygur half of each) can be applied for different race, gender and age periods. Moreover, it was observed that three metals (metalloids), As, Sc and Ba significantly varied between some subgroups. The established RVs will be valuable for future health or environmental evaluation, and can serve as a theoretical reference for RVs establishment in multi-race coexistence region.

Discovering significantly different metabolites between Han and Uygur two racial groups using urinary metabolomics in Xinjiang, China.

The main object of the study was to discover the associated significantly different metabolites between Han and Uygur, two main racial groups in Xinjiang, China with urinary metabolomics. Urine samples from 96 Han and 96 Uygur were analyzed using gas chromatography coupled to mass spectrometry (GCMS). Multivariate analysis was used to investigate the effect of race, age and gender on the urinary metabolomic profiles. Totally eight metabolites are identified contributed to the discrimination between Han and Uygur, including phenylacetylglutamine, myoinositol, d-galactose, ribonolactone, octadecanoic acid, galactitol, threonic acid and succinic acid. The metabolic pathways of them are mainly involved in carbohydrate, TCA cycle, fatty acid and mammalian gut microbial-related metabolism. Importantly, three metabolites, being used as biomarkers in clinic, are also differentially expressed in urine samples of two races. It suggests that the race effect should be critically considered prior to make diagnostic result in multi-race coexisted areas to decrease the false positive rate caused by above biomarkers. Moreover, the results show that the age-period and the gender also affect the urinary metabolomics profiles, but with different levels compared to race. We hope that the work can provide some help for developing novel diagnostic tests, understanding the mechanism of disease, designing clinical trials and refining precision medicine in multi-race coexisted areas.