Objective： To investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus. Methods：...Objective： To investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus. Methods： Ad-sCD40LIg- IRES2-CTLA4Ig, replication-defective adenovirus co-expressing sCD40LIg and CTLA4Ig, was constructed and identified. The co-expression of sCD40LIg and CTLA4Ig was evaluated with confocal laser scanning microscope and Western blotting. Skin transplantations of C57BL/6 to BALB/c mice were performed. PBS, Ad-Shuttle-CMV and Ad-sCD40LIg-IRES2-CTLA4Ig were administered. Skin graft survival was monitored and the mRNA expression of both genes was evaluated in the skin allografts. Results： Ad-sCD40LIg-IRESE-CTLA4Ig was constructed successfully and identified. The co-expression of sCD40LIg and CTLA4Ig was identified with confocal laser scanning microscopy and Western blotting. Compared to the skin graft mean survival time （MST） of non-treated group （（5.75±0.71） d） or Ad-Shuttle-CMV-treated group （（5.50±0.53） d）, the skin graft MST was dramatically prolonged in the Ad-sCD40LIg-IRESE-CTLA4Ig-treated group （（ 16.38± 1.19） d, P〈0.001）. The mRNA expression of both genes was detected. Conclusion： Ad-sCD40LIg-IRES2-CTLA4Ig, a replication-defective adenovirus carrying genes encoding sCD40LIg and CTLA4Ig, was constructed. Simultaneous blockade of CD40/CD40L and B7/CD28 costimulatory pathway mediated by replication-defective adenovirus significantly prolonged skin allografi survival in mice.展开更多
Skin grafting has been used as one of the most reliable tests to determine the genetic stability of laboratory animal such as mice and rats inbred line, but no identification of swine inbred lines by skin grafting has...Skin grafting has been used as one of the most reliable tests to determine the genetic stability of laboratory animal such as mice and rats inbred line, but no identification of swine inbred lines by skin grafting has been reported. At present, Wuzhishan miniature pig （WZSP） inbred line has acquired the F24 individuals in China. In order to verify whether WZSP inbred line had D^en cultivated successfully, allogeneic skin grafts and related research were performed on F20 individuals of WZSP inbreeding population, compared with a control group of autologous transplantation. We observed the transplant recipients＇ wounds, detected peripheral blood-related indicators interleukin-2, 4 and 10, CD4~ and CD8~ lymphocytes, and conducted hematoxylin-eosin （HE） and Masson＇s staining of skin to judge whether the immune rejection reactions occurred within 28 days after transplantation. Chr. 7 genomic heterozygosity of 48 WZSP individuals from F20 to F22 was analyzed by high-density single nucleotide polymorphism （SNP） chips （60 000 SNPs）. The result showed that there were no significant differences in graft skin, the plasma interleukin-2, 4, 10, CD4~ and CD8~, HE and Masson＇s staining results between the allograft and autograft groups, and no immune rejection occurred on the allograft group. We found that 11 genes in Chr. 7 of major histocompatibility complex （MHC） I and MHC II were homozygous which confirmed that immune antibody of the allograft and autograft groups were highly identical and also provided a theoretical basis to no immune rejection occurred on the allograft in the inbred WZSP. The result proved that the WZSP inbred line had been cultivated successfully for the first time in the world. The test methods also provide a scientific basis for the identification of swine and mammal inbred lines.展开更多
Full thickness skin grafts and composite grafts are a workhorse in reconstructive surgery of nose and ear defects whether they are originated from trauma, cancer surgery or burns. The reliability of these grafts has b...Full thickness skin grafts and composite grafts are a workhorse in reconstructive surgery of nose and ear defects whether they are originated from trauma, cancer surgery or burns. The reliability of these grafts has been proved with an established clinical use and morbidity to the donor site is minimal if harvesting and donor site selection if appropriate. Use of double or multiple grafts to reconstruct a complex defect of the nose has not been described and our report is meaningful for the fact that it describes a further use of the surgical concept of grafting.展开更多
Skin grafts may be used for coverage of facial defects in situations in which alternative methods of reconstruction,such as local flaps,are not an option.They may also be beneficial for patients who wish to avoid or w...Skin grafts may be used for coverage of facial defects in situations in which alternative methods of reconstruction,such as local flaps,are not an option.They may also be beneficial for patients who wish to avoid or who are not good candidates for more complex reconstruction.Full-thickness skin grafts often have a better color and texture match to adjacent skin when compared to split-thickness grafts;however,split-thickness grafts have lower metabolic demand and increased survival rate.Composite grafts may be very useful in the repair of defects with unique contour and support requirements,such as the nasal ala and eyelid.With all grafts,thoughtful planning and sound surgical technique are critical in achieving the best possible functional and aesthetic result.展开更多
基金Project (No. 30371416) supported by the National Natural ScienceFoundation of China
文摘Objective： To investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus. Methods： Ad-sCD40LIg- IRES2-CTLA4Ig, replication-defective adenovirus co-expressing sCD40LIg and CTLA4Ig, was constructed and identified. The co-expression of sCD40LIg and CTLA4Ig was evaluated with confocal laser scanning microscope and Western blotting. Skin transplantations of C57BL/6 to BALB/c mice were performed. PBS, Ad-Shuttle-CMV and Ad-sCD40LIg-IRES2-CTLA4Ig were administered. Skin graft survival was monitored and the mRNA expression of both genes was evaluated in the skin allografts. Results： Ad-sCD40LIg-IRESE-CTLA4Ig was constructed successfully and identified. The co-expression of sCD40LIg and CTLA4Ig was identified with confocal laser scanning microscopy and Western blotting. Compared to the skin graft mean survival time （MST） of non-treated group （（5.75±0.71） d） or Ad-Shuttle-CMV-treated group （（5.50±0.53） d）, the skin graft MST was dramatically prolonged in the Ad-sCD40LIg-IRESE-CTLA4Ig-treated group （（ 16.38± 1.19） d, P〈0.001）. The mRNA expression of both genes was detected. Conclusion： Ad-sCD40LIg-IRES2-CTLA4Ig, a replication-defective adenovirus carrying genes encoding sCD40LIg and CTLA4Ig, was constructed. Simultaneous blockade of CD40/CD40L and B7/CD28 costimulatory pathway mediated by replication-defective adenovirus significantly prolonged skin allografi survival in mice.
基金supported by the National High Technology Research and Development Program of China (2012AA020603)the National Transgenic Major Project, China (2008ZX08012-002-05)the National Key Technoligy R&D Program of China (2012BA13904)
文摘Skin grafting has been used as one of the most reliable tests to determine the genetic stability of laboratory animal such as mice and rats inbred line, but no identification of swine inbred lines by skin grafting has been reported. At present, Wuzhishan miniature pig （WZSP） inbred line has acquired the F24 individuals in China. In order to verify whether WZSP inbred line had D^en cultivated successfully, allogeneic skin grafts and related research were performed on F20 individuals of WZSP inbreeding population, compared with a control group of autologous transplantation. We observed the transplant recipients＇ wounds, detected peripheral blood-related indicators interleukin-2, 4 and 10, CD4~ and CD8~ lymphocytes, and conducted hematoxylin-eosin （HE） and Masson＇s staining of skin to judge whether the immune rejection reactions occurred within 28 days after transplantation. Chr. 7 genomic heterozygosity of 48 WZSP individuals from F20 to F22 was analyzed by high-density single nucleotide polymorphism （SNP） chips （60 000 SNPs）. The result showed that there were no significant differences in graft skin, the plasma interleukin-2, 4, 10, CD4~ and CD8~, HE and Masson＇s staining results between the allograft and autograft groups, and no immune rejection occurred on the allograft group. We found that 11 genes in Chr. 7 of major histocompatibility complex （MHC） I and MHC II were homozygous which confirmed that immune antibody of the allograft and autograft groups were highly identical and also provided a theoretical basis to no immune rejection occurred on the allograft in the inbred WZSP. The result proved that the WZSP inbred line had been cultivated successfully for the first time in the world. The test methods also provide a scientific basis for the identification of swine and mammal inbred lines.
文摘Full thickness skin grafts and composite grafts are a workhorse in reconstructive surgery of nose and ear defects whether they are originated from trauma, cancer surgery or burns. The reliability of these grafts has been proved with an established clinical use and morbidity to the donor site is minimal if harvesting and donor site selection if appropriate. Use of double or multiple grafts to reconstruct a complex defect of the nose has not been described and our report is meaningful for the fact that it describes a further use of the surgical concept of grafting.
文摘Skin grafts may be used for coverage of facial defects in situations in which alternative methods of reconstruction,such as local flaps,are not an option.They may also be beneficial for patients who wish to avoid or who are not good candidates for more complex reconstruction.Full-thickness skin grafts often have a better color and texture match to adjacent skin when compared to split-thickness grafts;however,split-thickness grafts have lower metabolic demand and increased survival rate.Composite grafts may be very useful in the repair of defects with unique contour and support requirements,such as the nasal ala and eyelid.With all grafts,thoughtful planning and sound surgical technique are critical in achieving the best possible functional and aesthetic result.
