Ting Cheng

Luminescent/Magnetic Hybrid Nanoparticles with Folate-Conjugated Peptide Composites for Tumor-Targeted Drug Delivery

We developed a novel chitosan-based luminescent/magnetic hybrid nanoparticles with folate-conjugated tetrapeptide composites (CLMNPs-tetrapeptide-FA) by conjugation in situ. First, chitosan, CdTe quantum dots (QDs), and superparamagnetic iron oxide were directly gelled into ternary hybrid nanogels. Subsequently, tetrapeptides (GFFG and LGPV) and folate were conjugated orderly into the hybrid nanoparticles. The morphology, composition, and properties of the as-prepared copolymers have also been characterized and determined using TEM, EDX, XRD, FTIR spectra, DLS, fluorescence spectroscopy, VSM, and fluorescence microscopy imaging studies. The size range of the end product CLMNPs-tetrapeptide-FA copolymers was from 150 to 190 nm under simulated physiological environment. In vivo, the experimental results of magnetic accumulation showed that the copolymers could be trapped in the tumor tissue under magnetic guidance. Under the present experimental conditions, the loading efficiencies of CPT were approximately 8.6 wt % for CLMNPs-GFFG-FA and 1.1 wt % for CLMNPs-LGPV-FA, respectively. The CPT cumulative release under dialysis condition mainly occurred for the first 28 h, and could reach 55% at pH 5.3 and 46% at pH 7.4 from CPT-loaded CLMNPs-GFFG-FA, and 69% at pH 5.3 and 57% at pH 7.4 from CPT-loaded CLMNPs-LGPV-FA within 28 h, respectively. The hemolysis percentages (<2%) and coagulation properties of blank and CPT-loaded copolymers were within the scope of safe values. Compared to free CPT, the CPT-loaded CLMNPs-tetrapeptide-FA copolymers showed specific targeting to A549 cells in vitro. More than 75% viability in L02 cells were seen in CLMNPs-GFFG-FA and CLMNPs-LGPV-FA copolymer concentration of 500 μg/mL, respectively. It was found that the two kinds of copolymers were transported into the A549 cells by a folate-receptor-mediated endocytosis mechanism. These results indicate that the multifunctional CLMNPs-tetrapeptide-FA copolymers possess a moderate CPT loading efficiency, low cytotoxicity, and favorable biocompatibility, and are promising candidates for tumor-targeted drug delivery.

Metabolic signatures of esophageal cancer: NMR-based metabolomics and UHPLC-based focused metabolomics of blood serum

Focused metabolic profiling is a powerful tool for the determination of biomarkers. Here, a more global proton nuclear magnetic resonance ((1)H NMR)-based metabolomic approach coupled with a relative simple ultra high performance liquid chromatography (UHPLC)-based focused metabolomic approach was developed and compared to characterize the systemic metabolic disturbances underlying esophageal cancer (EC) and identify possible early biomarkers for clinical prognosis. Serum metabolic profiling of patients with EC (n=25) and healthy controls (n=25) was performed by using both (1)H NMR and UHPLC, and metabolite identification was achieved by multivariate statistical analysis. Using orthogonal projection to least squares discriminant analysis (OPLS-DA), we could distinguish EC patients from healthy controls. The predictive power of the model derived from the UHPLC-based focused metabolomics performed better in both sensitivity and specificity than the results from the NMR-based metabolomics, suggesting that the focused metabolomic technique may be of advantage in the future for the determination of biomarkers. Moreover, focused metabolic profiling is highly simple, accurate and specific, and should prove equally valuable in metabolomic research applications. A total of nineteen significantly altered metabolites were identified as the potential disease associated biomarkers. Significant changes in lipid metabolism, amino acid metabolism, glycolysis, ketogenesis, tricarboxylic acid (TCA) cycle and energy metabolism were observed in EC patients compared with the healthy controls. These results demonstrated that metabolic profiling of serum could be useful as a screening tool for early EC diagnosis and prognosis, and might enhance our understanding of the mechanisms involved in the tumor progression.

Toxicity and biodistribution of aqueous synthesized ZnS and ZnO quantum dots in mice

Abstract In the present study, ZnS and ZnO quantum dots (QDs) were synthesized via an all-aqueous process with polyethylene glycol (PEG) chains on their surface, and their toxicity as well as biodistribution were evaluated. No haemolysis occurred at a high concentration of 1600 µg/mL in vitro haemolytic assay, which demonstrated that the QDs-PEG displayed good blood compatibility. Following intravenous administration at 2, 6, and 20 mg/kg of the QDs-PEG in mice, the biodistribution, excretion and biocompatibility were characterized at 1 h, 24 h and 7 days, respectively. Quantitative analysis results indicated that the biodistribution trend of ZnS QDs-PEG was similar to that of ZnO QDs-PEG. The QDs-PEG were mainly trapped in the lung and liver, and almost removed from blood within 1 h. QDs-PEG were primarily excreted in faeces at the 2 and 6 mg/kg doses. Coefficients, haematology, blood biochemistry and histopathology results indicated that the QDs-PEG were safe and biocompatible.

Rapid resolution liquid chromatography (RRLC) analysis of amino acids using pre-column derivatization.

A rapid resolution liquid chromatography (RRLC) method was developed for the simultaneous determination of 23 amino acids in rat serum after pre-column derivatization with 2,4-dinitrofluorobenzene (DNFB). The amino acid derivatives were separated on an Agilent Zorbax Eclipse Plus C(18) (4.6 mm×50 mm, 1.8 μm) column at 45°C. Ultraviolet (UV) detection was set at 360 nm. Good separation of 23 amino acids was achieved within 10 min with a ternary gradient elution of mobile phase at a flow rate of 1.5 mLmin(-1). Calibration curves were linear over the range from 1 to 500 μmolL(-1) with coefficients 0.9962 or better for each amino acid. The lower limits of quantification (LLOQ) of all 23 amino acids were 1μmolL(-1) with signal-to-noise (S/N) ratio ≥4. Intra- and Inter-day precisions, expressed as relative standard deviation (RSD) percentages, were ranged from 0.32% to 3.09% and 0.67% to 5.82%, respectively. Finally, it was successfully applied to the determination of amino acids in rat serum with recoveries ranged from 90.8% to 106.0% and RSD percentages ranged from 1.78% to 4.68%, respectively. The results showed that the proposed method provided a shorter elution time, better resolution and sharper peak shapes for all amino acids. Compared with the conventional high performance liquid chromatography (HPLC) methods, even some ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), the established RRLC method was superior performance.

Hepatotoxicity induced by ZnO quantum dots in mice

ZnO quantum dots (QDs) with unique optical properties are potential useful tools for biological labeling and biosensing. With the increasing use of ZnO QDs, the toxicity evaluation of ZnO QDs is urgent. In this study, the hepatotoxicity including serum aminotransferases (ALT and AST), antioxidant enzymes (CAT, GSH-Px and SOD), lipid peroxidation and ultrastructure were evaluated after consecutive intravenous injection of ZnO QDs and ZnO QDs–PEG for 7 days in mice. Both ZnO QDs and ZnO QDs–PEG did not affect the coefficient of liver and the levels of serum aminotransferases. The antioxidant enzymes and lipid peroxidation had significant change after injecting 5 mg kg−1 ZnO QDs in 24 h, but all of these parameters returned to control levels in 28 days. ZnO QDs–PEG had a less harmful effect on antioxidant enzymes and malondialdehyde than ZnO QDs at the same dose. According to the results of hepatocyte ultrastructure, both ZnO QDs and ZnO QDs–PEG were located in the mitochondrion and induced nuclear malformation in 24 h. The ultrastructure of hepatocyte was as normal as of the control group in 28 days and ZnO QDs were mainly trapped in the mitochondrion while ZnO QDs–PEG mainly accumulated in the lysosomes. These findings would be helpful for wide use of quantum dots based bioimaging and biomedical applications in the future.

High-efficiency extraction of nucleosides based on the combination of self-assembly ionic liquid layer and boronic acid-functionalized attapulgite

Boronate affinity materials have been widely used for selective capture of cis-diol-containing molecules, but most do not have sufficient extraction efficiency. We have prepared boronic acid-functionalized attapulgite, and then it was coated with imidazolium-based ionic liquid, 1-dodecyl-3-methylimidazolium bromide (C12mimBr), via physical self-assembly process. The extraction efficiency of the material increased dramatically after coated with C12mimBr, and its enrichment ratios for nucleosides increased by 9- to 282-fold. Besides, the C12mimBr-coated adsorbent did not lose selectivity and was able to capture cis-diols in the presence of a 100-fold excess of interferences. C12mimBr-coated material was applied to selective enrichment of nucleosides from human urine. The limits of detection and the limits of quantification were in the range of 0.06-0.46ngmL(-1) and 0.20-1.53ngmL(-1), respectively. Reproducibility of the method was determined with relative standard deviations≤9.9%. The recoveries of the target nucleosides from spiked human urine were in the range of 87.8-109.6%. In our preception, self-assembly ionic liquid layer can serve as a promising alternative to improve the extraction efficiency of boronate affinity materials.

Highly selective capture of nucleosides with boronic acid functionalized polymer brushes prepared by atom transfer radical polymerization.

The nucleoside or modified nucleoside level in biological fluids reflects the pathological or physiological state of the body. Boronate affinity absorbents are widely used to selectively extract nucleosides from complex samples. In this work, a novel functionalized absorbent was synthesized by attaching 4-mercaptophenylboronic acid to gold nanoparticles on modified attapulgite. The surface of the attapulgite was modified by poly(acryloyloxyethyltrimethyl ammonium chloride) by atom transfer radical polymerization, creating many polymer brushes on the surface. The resultant material exhibited superior binding capacity (30.83 mg/g) for adenosine and was able to capture cis-diol nucleosides from 1000-fold interferences. Finally, to demonstrate its potential for biomolecule extraction, this boronate affinity material was used to preconcentrate nucleosides from human urine and plasma.

Silica – Boronate affinity material for quick enrichment of intracellular nucleosides

Boronic acid modified materials have been widely used to adsorb nucleosides, but their adsorption capacities require further improvement. Most cis-diol containing biomolecules are in very low abundance along with interfering components in real samples, and need to be enriched specially. In this study, we synthesize a kind of silica absorbent modified with boronic acid derivative, using amorphous silica as raw material and obtaining high adsorption capacity for adenosine. In addition, the adsorption equilibrium can be completed within 10s and 1min for the desorption. Finally, the material was successfully applied to enrich nucleosides from cells and the spiked recoveries were found between 82.21% and 118.9%. The results showed that the prepared adsorbent has potential to effectively enrich cis-diol substances from cell samples.