A multilevel investigation to reveal the regulatory mechanism of lignin accumulation in juice sac granulation of pomelo.

Granulation of juice sacs is a physiological disorder, which affects pomelo fruit quality. Here, the transcriptome and ubiquitinome of the granulated juice sacs were analyzed in Guanxi pomelo. We found that lignin accumulation in the granulated juice sacs was regulated at transcription and protein modification levels. In transcriptome data, we found that the genes in lignin biosynthesis pathway and antioxidant enzyme system of the granulated juice sacs were significantly upregulated. However, in ubiquitinome data, we found that ubiquitinated antioxidant enzymes increased in abundance but the enzyme activities decreased after the modification, which gave rise to reactive oxygen species (ROS) contents in granulated juice sacs. This finding suggests that ubiquitination level of the antioxidant enzymes is negatively correlated with the enzyme activities. Increased H2O2 is considered to be a signaling molecule to activate the key gene expressions in lignin biosynthesis pathway, which leads to the lignification in granulated juice sacs of pomelo. This regulatory mechanism in juice sac granulation of pomelo was further confirmed through the verification experiment using tissue culture by adding H2O2 or dimethylthiourea (DMTU). Our findings suggest that scavenging H2O2 and other ROS are important for reducing lignin accumulation, alleviating juice sac granulation and improving pomelo fruit quality.

Effect of Extracellular Matrix Coating on Cancer Cell Membrane-Encapsulated Polyethyleneimine/DNA Complexes for Efficient and Targeted DNA Delivery In Vitro.

Polyethyleneimine (PEI) has a good spongy proton effect and is an excellent nonviral gene vector, but its high charge density leads to the instability and toxicity of PEI/DNA complexes. Cell membrane (CM) capsules provide a universal and natural solution for this problem. Here, CM-coated PEI/DNA capsules (CPDcs) were prepared through extrusion, and the extracellular matrix was coated on CPDcs (ECM-CPDcs) for improved targeting. The results showed that compared with PEI/DNA complexes, CPDcs had core-shell structures (PEI/DNA complexes were coated by a 6-10 nm layer), lower cytotoxicity, and obvious homologous targeting. The internalization and transfection efficiency of 293T-CM-coated PEI70k/DNA capsules (293T-CP70Dcs) were 91.8 and 74.5%, respectively, which were higher than those of PEI70k/DNA complexes. Then, the internalization and transfection efficiency of 293T-CP70Dcs were further improved by ECM coating, which were 94.7 and 78.9%, respectively. Then, the internalization and transfection efficiency of 293T-CP70Dcs were further improved by ECM coating, which were 94.7 and 78.9%, respectively. Moreover, the homologous targeting of various CPDcs was improved by ECM coating, and other CPDcs also showed similar effects as 293T-CP70Dcs after ECM coating. These findings suggest that tumor-targeted CPDcs may have considerable advantages in gene delivery.

Adsorbents prepared from epoxy-based porous materials of microcrystalline cellulose for excellent adsorption of anionic and cationic dyes.

It reported a porous material prepared from microcrystalline cellulose (MCC), to achieve rapid preparation of adsorbents. The porous material was characterized by several tools including 1H NMR, FTIR, XPS, and SEM. Two adsorbents were prepared and subjected to adsorption experiments. Dye adsorption experiments show that the adsorption driving is electrostatic interactions and the process is chemisorption. The maximum capacity of Microcrystalline cellulose-g-Poly (glycidyl methacrylate)-Tannins (MPT) reached 191.3 (Methylene blue), 123.7 mg g-1 (Rhodamine B), and Microcrystalline cellulose-g-Poly (glycidyl methacrylate)-Lysine (MPL) attained 425.8 (Methylene blue), 480.7 mg g-1 (Methyl orange). The results were followed the pseudo-second-order (PSO) and agreed with the Langmuir fit model. Adsorption-desorption cycling experiments further indicate that the adsorbent possesses outstanding reproducibility. At last, epoxidized bio-porous materials are positive in the preparation of dye adsorbents with critical adsorption properties.

Fabrication of zein-based hydrophilic nanoparticles for efficient gene delivery by layer-by-layer assembly.

As a natural biological macromolecule, zein has broad application prospects in drug delivery due to its unique self-assembly properties. In this work, zein/sodium alginate (Zein/SA) nanocomposites were prepared by a pH-cycle method, Then Zein/SA/PEI (ZSP) nanocomposites were prepared by efficient layer-by-layer assembly method, ZSP nanocomposite of higher transfection performance was further labeled by folic acid (FA). After characterizing the physicochemical properties of ZSP by various methods, the potential of ZSP as a gene delivery vehicle was explored in vitro. The results showed that ZSP had good dispersibility and stability, the diameter distribution was in the range of 124-203 nm, and it had a typical core-shell structure, which could effectively condensate DNA and protect it from nuclease hydrolysis. ZSP exhibited proton buffering capacity similar to PEI, lower cellular toxicity, lower protein adsorption and erythrocyte hemolysis effect than PEI. ZSP/pDNA complexes could be taken up by cells and exhibited higher transfection efficiency than PEI/DNA complexes at the same weight ratio. The transfection efficiency of the complex in HeLa and 293T cells can be improved by FA labeling, especially in HeLa cells. These results provide new perspective for the design and development of efficient zein-based gene delivery systems.

Therapeutic vaccination against leukaemia via the sustained release of co-encapsulated anti-PD-1 and a leukaemia-associated antigen.

Therapeutic leukaemia vaccines have shown modest potency. Here, we show that the co-encapsulation of a leukaemia-associated epitope peptide highly expressed in leukaemia patients and of the immune checkpoint inhibitor anti-programmed-cell-death-protein-1 (anti-PD-1) in degradable poly(lactic acid) microcapsules resulted in the sustained release of the peptide and of the antibody, which led to the recruitment of activated antigen-presenting cells to the injection site, their uptake of the peptide and the transportation of the anti-PD-1 antibody to lymph nodes, enhancing the expansion of epitope-specific T cells and the activation of cytotoxic T cells. After single subcutaneous injections of vaccine formulations with different epitope peptides, mice bearing leukaemia xenografts derived from humanized cell lines or from primary cells from patients showed better therapeutic outcomes than mice receiving repeated injections of free antigen, antibody and a commercial adjuvant. The sustained release of a tumour-associated peptide and of anti-PD-1 may represent a generalizable strategy for boosting antitumour immune responses to leukaemia.

TAT-functionalized PEI-grafting rice bran polysaccharides for safe and efficient gene delivery.

Polysaccharides are considered to be promising candidates for non-viral gene delivery because of their molecular diversity, which can be modified to fine-tune their physicochemical properties. In this work, transcriptional activator protein (TAT) functionalized PEI grafted polysaccharide polymer (PRBP) was prepared by using rice bran polysaccharide as the starting material, and characterized by various methods. The potential of TAT functionalized PRBP (PRBP-TAT) as gene vector was studied in vitro, including DNA loading capacity, DNA protection ability and biocompatibility. The cell uptake and transfection efficiency of the PRBP-TAT/pDNA polyplexes were studied. The results showed that PRBP-TAT could completely condense DNA at N/P 2. The PRBP-TAT/pDNA polyplexes could protect DNA from degrading by DNase and were efficiently internalized by cells. Biocompatibility result showed that PRBP-TAT had no significant cytotoxicity and effect on cell proliferation. At low N/P ratios of 1-3.5, PRBP-TAT showed higher transfection efficiency than PEI30k and PEI30k-grafted rice bran polysaccharide. PRBP-TAT and PEI showed the highest transfection efficiency of 42.8% and 28.1% when pDNA is 2 µg and N/P ratio is 1.5, respectively, while PRBP showed the highest transfection efficiency of 37.3% at N/P 2.5. These results indicate that PTA is a promising candidate vector for safe and efficient gene delivery.