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spersion of experimental results.In this study, the optimum human aFGF gene encoding haFGF135 was cloned in pET3c and transferred to Escherichia coli BL21(DE3) plysS. To enhance the yield of fermentation and the expression level of the target protein, the fermentation parameters, including temperature, pH, dissolved oxygen, glucose concentration, ammonium chloride concentration, induction time, and inducer (IPTG) concentration, were optimized. The optimized fermentation parameters were used in large-scale fermentation (30 L). Ion-exchange and heparin-affinity column chromatography techniques were used for separation and purification of rhaFGF135 protein. HPLC, isoelectric focusing electrophoresis, and mass spectrometry were used to detect the purity, isoelectric point, and molecular weight and peptide map of rhaFGF135 protein, respectively. Mitogenic activity of rhaFGF135 protein was detected in NIH-3T3 cells and a full-thickness injury wound diabetic rat model. The production and expression level of rhaFGF135 in the 30-L scale fermentation reached 80.4 ± 2.7 g/L culture and 37.8% ± 1.8%, respectively. The RP-HPLC and SDS-PAGE purity of the final rhaFGF135 product almost reached 100%, and the final pure protein yield was 158.6 ± 6.8 mg/L culture. Finally, the cell and animal experiments showed that rhaFGF135 retained a potent mitogenic activity. The large-scale process of rhaFGF135 production reported herein is relatively stable and time-saving, and thus, it can be used as an efficient and economic strategy for the synthesis of rhaFGF135 at the industrial level.The use of human cells for the construction of 3D organ models in vitro based on cell self-assembly and engineering design has recently increased in popularity in the field of biological science. Although the organoids are able to simulate the structures and functions of organs in vitro, the 3D models have difficulty in forming a complex vascular network that can recreate the interaction between tissue and vascular systems. Therefore, organoids are unable to survive, due to the lack of oxygen and nutrients, as well as the accumulation of metabolic waste. Organoids-on-a-chip provides a more controllable and favorable design platform for co-culture of different cells and tissue types in organoid systems, overcoming some of the limitations present in organoid culture. However, the majority of them has vascular networks that are not adequately elaborate to simulate signal communications between bionic microenvironment (e.g., fluid shear force) and multiple organs. Here, we will review the technological progress of the vascularization in organoids and organoids-on-a-chip and the development of intravital 3D and 4D bioprinting as a new way for vascularization, which can aid in further study on tissue or organ development, disease research and regenerative medicine.The drumstick tree is a fast-growing multipurpose tree with a large biomass and high nutritional value. However, it has rarely been exploited as a protein source. This study investigated solid-state fermentation induced by Aspergillus niger, Candida utilis and Bacillus subtilis to obtain high-quality protein feed from drumstick leaf flour. The results showed that fermentation induced significant changes in the nutritional composition of drumstick leaf flour. The concentrations of crude protein, small peptides and amino acids increased significantly after fermentation. The protein profile was also affected by the fermentation process. Macromolecular proteins in drumstick leaf flour were degraded, whereas other high molecular weight proteins were increased. However, the concentrations of crude fat, fiber, total sugar and reducing sugar were decreased, as were the anti-nutritional factors tannins, phytic acid and glucosinolates. After 24 h fermentation, the concentrations of total phenolics and flavonoids were increased. The antioxidant capacity was also significantly enhanced.Co-digestion of fats, oils, and grease (FOG) with food waste (FW) can improve the energy recovery in anaerobic membrane bioreactors (AnMBRs). Here, we investigated the effect of co-digestion of FW and FOG in AnMBRs at fat mass loading of 0.5, 0.75, and 1.0 kg m-3 day-1 with a constant organic loading rate of 5.0 gCOD L-1 day-1 in both a single-phase (SP) and two-phase (TP) configuration. A separate mono-digestion of FW at an identical organic loading rate was used as the benchmark. During co-digestion, higher daily biogas production, ranging from 4.0 to 12.0%, was observed in the two-phase methane phase (TP-MP) reactor compared to the SP reactor, but the difference was statistically insignificant (p > 0.05) due to the high variability in daily biogas production. However, the co-digestion of FW with FOG at 1.0 kg m-3 day-1 fat loading rate significantly (p less then 0.05) improved daily biogas production in both the SP (11.0%) and TP (13.0%) reactors compared to the mono-digestion of FW. Microbial community analyses using cDNA-based MinION sequencing of weekly biomass samples from the AnMBRs revealed the prevalence of Lactobacillus (92.2-95.7% relative activity) and Anaerolineaceae (13.3-57.5% relative activity), which are known as fermenters and fatty acid degraders. Syntrophic fatty acid oxidizers were mostly present in the SP and TP-MP reactors, possibly because of the low pH and short solid retention time (SRT) in the acid phase digesters. A greater abundance of the mcrA gene copies (and methanogens) was observed in the SP and MP reactors compared to the acid-phase (AP) reactors. This study demonstrates that FW and FOG can be effectively co-digested in AnMBRs and is expected to inform full-scale decisions on the optimum fat loading rate.Drug delivery systems have good biocompatibiliy and low side effects for cancer treatment, but overcoming high efficiency of drug-loading and the drug-targeting controlled release still remains challenging. Selleckchem MELK-8a In this work, supramolecular vesicles, with pH-triggering effect, have been successfully constructed for drug delivery, which are fabricated by the complexation between a cationic pillar[5]arene (DAWP5) and a sodium dodecyl sulfonate (SDS) in aqueous solution. Drug-loading and releasing results demonstrated that anticancer drug doxorubicin (DOX) could be loaded efficiently by such cationic vesicles in neutral condition, and the drug release could be controlled in the simulated weak acid environment of tumor cells. Moreover, the vesicles had low cytotoxicity to normal human cell (L02), while the DOX-loaded vesicles could significantly enhance the cytotoxicity of free DOX for normal cell L02 and four tested tumor cells (Hela, HepG2, MGC-803 and T24). Especially for HepG2, after 24 h incubation time, IC50 of DOX-loaded vesicles was only 0.