ABSTRACT
Objective: To evaluate the advantages of microskin graft using acellular porcine skin for treatment of extensive deep burns by comparing with that using allogeneic skin. Methods: A retrospective analysis was conducted on 70 severe burn patients who were treated in the Second Affiliated Hospital of Kunming Medical University during Jan. 1999 to Jan. 2011. The patients were divided into the acellular porcine skin group and allogeneic skin group, each containing 35 patients. The survival rates of microskin grafts were determined at 4 weeks post-operation. Besides, the rejection of acellular porcine skin and allogeneic skin, changes of body temperature, white blood cell (WBC) count, lymphocyte and serum protein were observed at pre- and post-operation. Results: (1) The survival rate was (71. 5 ± 6. 6)% in acellular porcine skin group and (70. 6 ± 7. 5)% in allogeneic skin group, with no significant difference found between the two groups (P>0. 05). (2) Acellular porcine skin group. At 3 days post-operation the acellular porcine skin was still attached to the wound, most of the skin was not discolorated, and small part of the skin became cinnamomeous. The acellular porcine skin was gradually separated from the auto-microskin at 3-4 weeks post operation, and there was small amount of exudates under the acellular porcine skin, which could be drained through a small cut. In the pressed area, there was still a small amount of exudates, but the acellular porcine skin was not dissolved and the microskin grafts survived and became confluent. (3) Allogeneic skin group. The allogeneic epidermal was rejected and was off from the wound at 3-14 days post transplantation, and at 10-30 days after transplantation the allogenic dermis became dry. During 25-60 days after transplantation, the allogenic dermis was completely stripped off, the microskin grafts became confluent, and the wound was healed. (4) The body temperature of the two groups was significantly descended after operation (P0. 05). Conclusion: Microskin graft using acellular porcine skin, instead of allogeneic skin, for extensive burn patients can inhibit systematic inflammatory response, improve the nutrition condition, and reduce the using of allogeneic skin. Acellular porcine skin might be a suitable alternative for allogeneic skin.
ABSTRACT
OBJECTIVE: To construct recombination eukaryote expression plasmid for human parathyroid hormone (PTH) gene, assay PTH expression and biological activity after transfection in vitro and evaluate gene therapy effect on hypoparathyroidism (HPT). METHODS: (1) PTH gene was amplified from human embryonic parathyroid gland tissue, and plasmid pcDNA3.1-PTH-GFP (pcDPG) was constructed by TOPO recombination technique. Digestion, PCR and sequencing were used to identify the positive vectors. (2) pcDPG was transformed into 293 cells by Lipofectamine 2000(TM), fluorescent inverted microscope was used to observe GFP expression, and PTH gene expression was assayed by RT-PCR technique. (3) PTH protein in supernatant was purified and evaluated biological activity. (4) HPT rabbit models were developed and plasmid pcDPG was injected in skeletal muscles, respectively. Serum calcium, phosphonium and PTH were assayed and pathological changes observed. RESULTS: (1) The findings in digestion and PCR were accorded to anticipation and sequences in report were identified to reference at 99.30%. (2) 24 h after transfection GFP expression could be detected and enhanced with time prolonged arriving to 38.91% and 62.45% at 48 h. PTH gene expression could be detected by RT-PCR. (3) Purified PTH protein made the signs of HPT disappear. (4) Serum calcium and PTH levels were lower than those of pre-operation (P < 0.05) and serum phosphonium enhanced to normal standard 48 h after treatment at the plasmid pcDPG doses of 300 microg/kg and 500 microg/kg. CONCLUSION: Recombination plasmid pcDPG was transformed effectively in vitro and the transfected cells produced PTH protein with biological activity. Besides, the satisfactory therapeutic effect of HPT rabbits was attained by pcDPG plasmid, which provided a foundation for further study of HPT gene therapy.
