Immunomagnetic capture and traceless release of native tumor-derived exosomes from human plasma for exploring interaction with recipient cells by aptamer-functionalized nanoflowers.

BACKGROUND: Tumor-derived exosomes (TEXs) play an important role in the development process of cancer, which can transport a large number of carcinogenic molecules to normal cells, and subsequently promote tumor metastasis. However, TEXs that were utilized in most of previous researches were obtained from the cell medium of tumor cell lines, which cannot reflect the physiological state of primary cells in vivo. Isolation of native TEXs from human plasma with intact function is contributed to exploring the interaction between TEXs and recipient cells for understanding their true biological functions. RESULTS: We developed a strategy that involves both capture and release processes to obtain native TEXs from plasma of cancer patients. An MoS2-based immunomagnetic probe (Fe3O4@MoS2-Au-Aptamer, named as FMAA) with the advantages of high surface area, magnetic response and abundant affinity sites was designed and synthesized to capture TEXs through recognizing high-expression tumor-associated antigens of EpCAM. With the assistance of complementary sequences of EpCAM, TEXs were released with non-destruction and no residual labels. According to NTA analysis, 107-108 TEXs were recovered from per mL plasma of breast cancer patients. The interaction between native TEXs and normal epithelial cells confirms TEXs could induce significant activation of autophagy of recipient cells with co-culture for 12 h. Proteomics analysis demonstrated a total of 637 proteins inside epithelial cells had dynamic expression with the stimulation of TEXs and 5 proteins in the pathway of autophagy had elevated expression level. SIGNIFICANCE: This work not only obtains native TEXs from human plasma with non-destruction and no residual labels, but also explores the interaction between TEXs and recipient cells for understanding their true biological functions, which will accelerate the application of TEXs in the field of biomarkers and therapeutic drugs.

Pollen-like silica nanoparticles as a nanocarrier for tumor targeted and pH-responsive drug delivery.

Aptamer modified hollow silica nanoparticles with pollen structure (plSP@aptamer) were synthesized and used as a nanocarrier for tumor targeted and pH-responsive drug delivery. The 292 ± 14 nm interior void in diameter together with 11.8 nm surface pore size of plSP@aptamer nanoparticles contributed to a high drug loading efficiency of 0.509 g g-1. Furthermore, the drug delivery system was pH-responsive, and the releasing efficiency was up to 87.5% at pH of 5. The special spikes of this plSP@aptamer nanoparticles acted as "entry claws" to enhanced the interaction between cell and drug nanocarriers and then increased the internalization rate of drug vehicles. The cell uptake assay suggested that most of doxorubicin (DOX)@plSP@aptamer nanoparticles can escape form lysosome and located in nuclei of MCF-7 cells. The targeted performance testing showed that almost no DOX@plSP@aptamer were internalized by normal cells, indicating a high specificity of our drug vehicles. The cytotoxicity of nanoparticles was also investigated, the plSP@aptamer particles had excellent biocompatibility and the cell viability was nearly 100%. After loaded with DOX, DOX@plSP@aptamer showed great potential in targeted therapy of tumors, and only 4.2% MCF-7 cells were viable.

A novel double-component MOAC honeycomb composite with pollen grains as a template for phosphoproteomics research.

The enrichment and separation of phosphopeptides from mixed biological samples is a technologically very significance, but highly challenging work. Current designed materials are mainly based on the broad and effective adsorptive character of metal oxide affinity chromatography (MOAC). Though significant progress has been made in the enrichment of phosphopeptides with MOAC material, there are chances for further development. In this study, a novel pollen-based MOAC honeycomb material was firstly explored in which the suitable hydrophilic channels preferentially enrich much more endogenous phosphopeptides than nonphosphopeptides or proteins while doping binary metal oxides at the atomic level and the ultra-high specific surface area have further allowed it to possess more effective active sites. Based on these unique features, the pollen-based material exhibited high selectivity for ß-casein (mass ratio of ß-casein/BSA, 1:1500), ultra-low detection limit (0.1fmol), desirable reusability. Moreover, the bionics MOAC composites were investigated in the enrichment of phosphopeptides from nonfat milk, human serum (male and female at the same age) and mice liver, results of which indicate the great potential of the composite for the phosphoproteome analysis of complex biological samples through the cheap and environmentally friendly process.

The effects of low-intensity pulsed ultrasound and nanomagnet applications on the expressions of MMP-13 and MAPKs in rabbit knee osteoarthritis.

An animal model of Osteoarthritis (OA) was established to observe the influences of low-intensity pulsed ultrasound (LIPUS) and nano magnet application (NMA) on Collagenase 3 (MMP-13) expression and the activation status of mitogen activated protein kinases (MAPKs) in rabbit. 24 experimental rabbits from New Zealand were randomly divided into four groups: LIPUS, NMA, LIPUS + NMA group, and control group. The experimental rabbit OA model was established in the right knee joint of rabbits received ACLT operation. Rabbits in LIPUS group received LIPUS treatment and rabbits in NMA group were given NMA treatment. In LIPUS + NMA group, both treatments were applied on experimental rabbits everyday. However, the rabbits in control group only underwent ACLT operation. Four weeks later all rabbits were killed and changes of histopathology in rabbit articular cartilage were assessed and evaluated using Mankin method (Modified Mankin Scale). The protein expressions of MMP-13 and MAPKs were estimated using Western Blot. The results showed that both LIPUS and NMA treatments could significantly decrease the Mankin scores and suppress the expression level of MMP-13. However, there were some inverse results of MAPKs expression in these two applications and imply their treatment mechanisms of OA were different from each other.

High throughput identification of components from traditional Chinese medicine herbs by utilizing graphene or graphene oxide as MALDI-TOF-MS matrix.

In this work, graphene or graphene oxide was utilized, for the first time, to identify small molecular components from traditional Chinese medicine (TCM) herbs, by acting as matrix of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Due to the large surface area of graphene or graphene oxide, the analytes were trapped tightly to the matrix, which avoids the contamination of the ion source and vacuum system. Besides, their excellent electronic, thermal and mechanical properties make them desired matrices for MALDI-TOF-MS. Stable analysis was achieved with no background inference even at the concentration of 100 nM. Moreover, the limit of detection (LOD) could be greatly lowered by utilizing graphene or graphene oxide as a pre-enrichment adsorbent. In summary, the promoted MALDI-TOF-MS methodology was demonstrated to be simple, sensitive, fast, cost effective and, most importantly, high throughput.

Recent developments and contributions from Chinese scientists in multidimensional separations for proteomics and traditional Chinese medicines.

The most basic task in proteomics remains the detection and identification of proteins from a biological sample, and the most traditional way to achieve this goal consists in protein separations performed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Yet the 2-D PAGE-mass spectrometry (MS) approach has its drawbacks with regard to automation, sensitivity, and throughput. Consequently, considerable effort has been devoted to the development of non-gel-based proteome separation technologies in an effort to alleviate the shortcomings of 2-D PAGE. In addition, traditional Chinese medicines (TCMs), due to their long period of clinical testing and reliable therapeutic efficacy, are attracting increased global attention. However, hundreds or even thousands of components are usually present in TCMs, which results in great difficulties of separation. As a mainstream separation tool, multidimensional liquid separation systems have shown powerful separation ability, high peak capacity, and excellent detectability in the analysis of complex samples including biological samples and TCMs, etc. Therefore, this review emphasizes the most recent advances in multidimensional liquid chromatography and capillary electrophoresis-based separation techniques, and the corresponding applications in proteomics and TCMs. In view of the significant contributions from Chinese scientists, this review focuses mainly on the work of Chinese scientists in the above fields.

Recent developments in sample preparation techniques for chromatography analysis of traditional Chinese medicines.

Traditional Chinese medicines (TCMs) have a long history dating back thousands of years. Recently, there has been increasing interest worldwide in the use of TCMs for the prevention and treatment of various illnesses. In China, a large number of analytical tools, especially chromatographic techniques have been used to analyze the constituents of TCMs in order to control their quality and discover new bioactive compounds. In this paper, recent developments in sample preparation techniques for the extraction, clean-up, and concentration of analytes from TCMs are compared. These techniques include headspace solid-phase microextraction (HS-SPME), headspace liquid-phase microextraction (HS-LPME), microwave-assisted extraction (MAE), supercritical-fluid extraction (SFE), pressurized-liquid extraction (PLE), and microwave distillation (MD).