Construction of human recombinant lentiviral vector of GP73and its expression in DC2.4 cells

Objective:This study aimed to prepare Golgi protein-73(GP73)-DC vaccine by constructing human lentiviral vector of GP73and transfecting this vector in vitro into a murine dendritic cell line DC 2.4.This will lay the foundation for further study on targeted therapy of hepatocellular carcinoma using GP73-DC vaccine.Methods:Human GP73 gene was cloned into lentivirus GV358vector and validated by polymerase chain reaction(PCR)and sequencing.The validated GV358-GP73vector was then cotransfected with pHelper 1.0plasmid into 293Tcells,in order to prepare lentivirus GP73vector.Finally,the protein expression of GP73 was achieved by transfecting the lentivirus GP73vector into DC 2.4cells in vitro and determined by Western blotting.Results:PCR and sequencing confirmed that the GV358-GP73lentivirus vector was constructed correctly,and the titer level reached 2×108TU/mL.After transfection in DC 2.4cells,the GP73protein level was significantly enhanced.Conclusion:The lentivirus GV538-GP73vector was constructed successfully,and could be effectively expressed in DC 2.4cells.

An Injectable Hydrogel with or without Drugs for Prevention of Epidural Scar Adhesion after Laminectomy in Rats

Epidural scarring occurs inevitably in the defect after spinal laminectomy, and thus how to prevent or reduce it becomes a challenging topic. In the present study, an injectable hydrogel and its dexamethasone （DEX）-loaded hydrogel systems were adopted to prevent epidural scarring in a postlaminectomy rat model. The hydrogel system composed of poly（D,L-lactic acid-co-glycolic acid）-poly（ethylene glycol）-poly（D,L-lactic acid-co-glycolic acid） （PLGA-PEG-PLGA） triblock copolymers was a free-flowing sol at room temperature, and spontaneously turned into a semi-solid gel at body temperature. A lumbar 3 total laminectomy was performed on Sprague Dawley （SD） rats, and the efficacy of the injectable hydrogel with or without drugs in preventing epidural scar formation was evaluated via the gross anatomical observation and histological examination at one month post-surgery. The results demonstrate that the use of hydrogel alone reduced epidural scarring significantly, whereas the efficacy of the DEX-loaded hydrogels presented an irregular dose-dependency of drug and even the inappropriate drug doses resulted in the negative results. Therefore, the present study confirms that the PLGA-PEG- PLGA hydrogel holds potential as a barrier device to decrease peridural scarring, and reveals that the sustained delivery of the steroid hormone DEX to prevent surgery-related adhesions in the laminectomy defect is complex. Moreover, our in vivo studies also remind the researchers to pay attention to the irregular dose-dependency of the hormone drugs.