Crowleymann0683
Importantly, the secondary structure of the protein was retained after purification. Further, this recombinant PDX1 fusion protein was applied to human cells and showed the ability to enter the cells as well as translocate to the nucleus. This recombinant tool can be used as a safe tool and can potentially replace its genetic and viral forms in the reprogramming process to induce a β-cell-specific transcriptional profile in an integration-free manner. Additionally, it can also be used to elucidate its role in cellular processes and for structural and biochemical studies.The gene encoding the phage major capsid protein 10A was cloned into the prokaryotic expression vector pET24a, and a 6XHis-tag was fused to the 3'-end of the 10A gene to verify complete expression. The recombinant plasmid was transformed into Escherichia coli (E. coli) BL21 (DE3) cells, and 10A expression was induced by IPTG. SDS-PAGE and Western blot were used to confirm the target protein expression. The T7Select10-3b vector was added to the cultured bacteria expressing 10A at a multiplicity of infection (MOI) ranging from 0.01 to 0.1, and complete lysis of the bacteria was monitored by absorbance changes in the medium. The recombinant phage (reP) was harvested by PEG/NaCl sedimentation and resuspended in PBS. ELISA was performed to verify the presence of the 6XHis-tag on the surface of reP. The 10A-fusion expression vectors (pET10A-flag, pET10A-egfp, and pET10A-pct) were constructed, and fusion proteins were expressed and detected by the same method. The corresponding rePs (reP-Flag, reP-EGFP, and reP-PCT) were prepared by T7Select10-3b infection. After the expression of the peptides/proteins on the reP surfaces was confirmed, reP-Flag and reP-PCT were used to immunize mice to prepare anti-Flag and anti-PCT antibodies. The results showed that rePs prepared using the 10A-fusion vector and T7Select10-3b can be used as antigens to immunize mice and prepare antibodies. This method may be able to meet the rapid antigen preparation requirements for antibody production. Notably, the recombinant phage (reP) described in this study was obtained by the sedimentation method from T7Select10-3b-infected E. coli BL21 (DE3) cells carrying the major capsid protein 10A expression vector or 10A-fusion protein vector.In situ-forming injectable hydrogels are smart biomaterials that can be implanted into living bodies with minimal invasion. Due to pioneer work of Prof. Sung Wan Kim in this field, injectable hydrogels have shown great potentials in many different biomedical applications. Biodegradable and injectable hydrogels can be administered at room temperature as viscous polymer sols. They will degrade after accomplishing their tasks. Before injecting into living bodies, active substances can be loaded into viscous polymer sols with a high loading efficiency by simple mixing. After injecting into living bodies, active substances-loaded hydrogels can be formed and active substances can be released in a controlled manner upon diffusion or polymer degradation. Due to their outstanding properties and unique features, injectable hydrogels are very promising in many biomedical applications including drug/protein/gene delivery, tissue engineering, and regenerative medicine. In this review, we briefly introduce recent development of several important types of in situ-forming injectable hydrogels reported by our group during the last three years. Important properties and potential applications (such as cancer therapy, insulin release and wound healing) of these injectable hydrogels are reviewed. Challenges and perspectives in this research field are also discussed.Thrombosis and inflammation after implantation remain unsolved problems associated with various medical devices with blood-contacting applications. In this study, we develop a multifunctional biomaterial with enhanced hemocompatibility and anti-inflammatory effects by combining the anticoagulant activity of heparin with the vasodilatory and anti-inflammatory properties of nitric oxide (NO). selleck kinase inhibitor The co-immobilization of these two key molecules with distinct therapeutic effects is achieved by simultaneous conjugation of heparin (HT) and copper nanoparticles (Cu NPs), an NO-generating catalyst, via a simple tyrosinase (Tyr)-mediated reaction. The resulting immobilized surface showed long-term, stable and adjustable NO release for 14 days. Importantly, the makeup of the material endows the surface with the ability to promote endothelialization and to inhibit coagulation, platelet activation and smooth muscle cell proliferation. In addition, the HT/Cu NP co-immobilized surface enhanced macrophage polarization towards the M2 phenotype in vitro, which can reduce the inflammatory response and improve the adaptation of implants in vivo. This study demonstrated a simple but efficient method of developing a multifunctional surface for blood-contacting devices.A series of novel myricetin derivatives containing benzimidazole skeleton were constructed. The structure of compound 4g was further corroborated via X-ray single crystal diffractometer. The antimicrobial bioassays showed that all compounds exhibited potent inhibitory activities against Xanthomonas axonopodis pv. Citri (Xac), Ralstonia solanacearum (Rs) and Xanthomonas oryzae pv. Oryzae (Xoo) in vitro. Significantly, compound 4q showed the best inhibitory activities against Xoo, with the EC50 value of 8.2 μg/mL, which was better than thiodiazole copper (83.1 μg/mL) and bismerthiazol (60.1 μg/mL). In vivo experimental studies showed that compound 4q can treat rice bacterial leaf blight at 200 μg/mL, and the corresponding curative and protection efficiencies were 45.2 and 48.6%, respectively. Meanwhile, the antimicrobial mechanism of the compounds 4l and 4q were investigated through scanning electron microscopy (SEM). Studies showed that compounds 4l or 4q can cause deformation or rupture of Rs or Xoo cell membrane. These results indicated that novel benzimidazole-containing myricetin derivatives can be used as a potential antibacterial reagent.One of the important problems in oncology is finding the genes that perturb the cell functionality and cause cancer. These genes, namely cancer driver genes (CDGs), when mutated, lead to the activation of the abnormal proteins. This abnormality is passed on to other genes by protein-protein interactions, which can cause cells to uncontrollably multiply and become cancerous. So, many methods have been introduced to predict this group of genes. Most of these methods are computational-based, which identify the CDGs based on mutations and genomic data. In this study, we proposed KatzDriver, as a network-based approach, in order to detect CDGs. This method is able to calculate the relative impact of each gene in the spread of abnormality in the gene regulatory network. In this approach, we firstly create the studied networks using gene expression and regulatory interaction data. Then by combining the topological and biological data, the weights of edges (regulatory interactions) and nodes (genes) are calculated. Afterward, based on the KATZ approach, the receiving and broadcasting powers of each gene were calculated to find the relative impact of each gene.