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Objective:To investigate the effect of the new nanomaterials,polyamidoamine dendrimers PAMAM-D,mediated foreign gene human decay accelerating factor(hDAF) for xenotransplantation transfected pig sperm cells.Methods:PAMAM-D/hDAF cDNA compounds were made.The compounds were divided into 0.2 μg,0.4μg,0.6μg,0.8 μg and 1.0μg groups (each group adding corresponding dose of PAMAM-D in accordance with the N/P ratio 10:1,20:1,40:1),then were digested by restriction enzymes.The compounds were incubated with washed pig sperm cells.Then the transfection efficiency was detected by in situ hybridization in the different groups.Results:The PAMAM-D molecule can prevent electrophoretic migration of DNA in the compound.After digested the compounds by restriction enzymes,DNA can not be degraded.The transfection efficiency was different in different groups.Among the total,the efficiency was higher in both groups of 0.4 μg and 0.6μg than that of others.The top was the group of 0.4 μg linear plasmid plus PAMAM-D when the N/P ratio was 20:1(47.5%±O.2%,167% vs control group).Conclusion:PAMAM-D can improve the efficiency of exogenous gene transfeeted pig sperm cells,which can reinforce the stable binding of exogenous DNA to sperm cells.
ABSTRACT
Objective To construct transgenic mice tissue-specifically expressing human decay accelerating factor (DAF) on the vascular endothelium for xenotransplantation. Methods Transgenic mice expressing hDAF were generated by microinjection of a hDAF-minigene under the control of the human intercellular adhesion molecule-2 (ICAM-2) promoter. PCR analysis and Southern blot analysis of genomic DNA were used to examine the presence of the transgene in the genome of the offspring, and the expression was detected by RT-PCR and flow cytometry respectively. Immunohistochemical assay was performed to detect the distribution of hDAF on the tissues from these transgenic mice. An ex vivo perfusion model was used to compare hearts from these hDAF mice with wild-type hearts.Results After microinjection of gene, 21 of 133 mice born were shown to be transgenic. Human DAF gene was expressed on the surface of leucocytes in 8 of the 21 hDAF-integrated mice. Expression levels of 8 founders ranged from 70 % to 95 % of that on human leucocytes. Human DAF were strongly expressed on the vascular endothelium of heart, kidney and liver, with little or no positive staining observed on non-endothelial cells. These endothelial-specific hDAF hearts displayed prolonged survival compared to wild-type hearts during perfusion with 20 % human plasma and maintained approximately 20 % maximum work. Conclusions The introduced hDAF gene has been specifically expressed on the vascular endothelium of transgenic mice. The hDAF tissue-specifically expressed on vascular endothelium could enhance xenograft survival.
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Objective To investigate the causes,diagnosis and management of Wunderlich syndrome. Methods A total of 19 cases (15 men and 4 women) of Wunderlich syndrome were reported.Their average age was 47 years (range,11-75 years).Imaging examinations (B-ultrasound,IVU,CT,arteriography,MRI) together with laboratory testing and history supported the etiologic diagnosis of tumorous disease (n=10),non-tumorous disease (n=7) and unknown causes (n=2).Surgical management was performed in 9 cases of kidney tumor and 1 of ureter stone with obstructive hydronephrosis.Conservative treatment was adopted in the rest 9 cases.In 2 of the conservatively treated cases,the bleeding artery was embolized because of persistent bleeding. Results In the 9 tumor cases undergoing surgery,pathology confirmed renal cell carcinoma in 7,malignant fibrous histocytoma in 1 and transitional cell carcinoma of the renal pelvis in 1.These 9 cases all recovered after operation and experienced follow-up for 6 months to 3 years.Of them,5 cases died of tumor recurrence and metastasis;3 survived disease-free for 9 months to 2 years;1 had liver metastasis 1 year after operation and survived with tumor for 11 months.One case of ureter stone got significant improvement.The 9 cases undergoing conservative treatment had satisfactory results. Conclusions The main cause of Wunderlich syndrome is renal tumor.The conservative approach appears to be the most acceptable option for the patients with Wunderlich syndrome who have no malignant tumorous cause.