Objective To observe the effectiveness of transferring human A20 gene into endothelial cells. Methods The shuttle plasmid pCAGGSEHA20 was constructed using gene cloning and recombined technique. Endothelial cells were transfected with pCAGGSEHA20 and pMAMneo by DOTAP. The postive cell clones were selected with G418. The stable transfection and expression of A20 in the endothelial cells were determined by in situ hybridization and immunohistochemical analysis. Results The two fragments digested from pCAGGSEHA20 by EcoRⅠ represented 4 6?kb and 2 3?kb by electrophoresis, which were confirmed to be the carrier and the A20 gene fragments inserted originally. The above results indicate that the construction of pCAGGSEHA20 was successful. Abundant A20 stable expression in endothelial cells transfected with pCAGGSEHA20 was confirmed by in situ hybridization and immunohistochemical analysis.Conclusion By means of the DOTAP, hA20 gene can be transferred and stably expressed in endothelial cells.

Objective Previous studies have demonstrated that LPS can induce endothelial cell activation and the expression of E-selectin. In this study, we examined whether A20 gene could inhibit the expression of E-selectin in endothelial cells induced by LPS. Methods With the help of DOTAP, endothelial cells were transfected with pCDNA3.1 EHA20. The postive cell clones were selected with G418.The stable transfection and expression of A20 in the endothelial cells were determined by immunofluorescence analysis. The E-selectin expression was checked by immunofluorescence, Western blot and in situ hybridization. Results Abundant A20 stable expression in endothelial cells transfected with pCDNA3.1 EHA20 was confirmed by immunofluorescence analysis. E-selectin expression increased in LPS-inducible endothelial cells. A20 gene inhibited 90% LPS-inducible E-selectin expression(P

Objective To investigate the effects of zinc finger protein gene A20 on the inhibition of lipopolysaccharide (LPS) induced interleukin 8 (IL 8) expression in endothelial cells. Methods Plasmid pcDNA3.1EHA20 was transfected into human umbilical vein endothelial cells (HUVECs) by DOTAP method. The positive cell clones were screened with G418. The stable transfection and expression of A20 in HUVECs were determined by immunofluorescent analysis. IL 8 expression was detected by sandwich ELISA with double monoclonal antibodies. Results High expression of A20 gene in HUVECs transfected with pCDNA3.1EHA20 was confirmed by immunofluorescent analysis. IL 8 expression increased in LPS induced endothelial cells. A20 gene could inhibit more than 70% LPS induced IL 8 expression ( P

Objective To construct the recombinant adenovirus of mouse Osf2/Cbfal gene and to observe its ability to infect NIH3T3 fibroblasts. Methods The Osf2/Cbfal gene fragment was cloned into the shuttle plasmid pAdTrack-CMV to form the transfer vector by the method of homogenous recombination in bacteria. Then the recombinant adenovirus was transfected into NIH3T3 cells using Lipofectine DOTAP, The target gene was detected by poly-merase chain reaction (PCR). The titer and its infection rate were determined using the green fluorescent protein (GFP) expression in the shuttle plasmid. Results Restriction endonuclease and PCR analyses confirmed that the Osf2/Cbfal gene was successfully inserted into the adenovirus vector. The titer of the recombinant adenovirus was 1.6?1012 pfu/ml. The adenovirus had a strong effect on NIH3T3 cells. Conclusion The recombinant adenovirus containing Osf2/Cbfal gene was successfully constructed by the method of homogenous recombination in bacteria.

Objective To explore the experiences of the diagnosis and treatment of urothelial tumor in multiple organs.Methods Clinical data of 10 patients with urothelial tumor in multiple organs were retrospectively reviewed.Urothelial tumors were found in two or more organs at the same time by B ultrasound,IVU,R-P,CTU,MRU,cystoscopy,ureteroscopy and so on before operation.Results 6 cases were operated by radical total nephroureterectomy and partial cystectomy,3 cases were operated by radical total nephroureterectomy and cystectomy with urinary diversion,1 case was operated by partial ureterectomy and total cystectomy.8 of them were alive,1 case was operated by total urethrectomy because of tumor recurrence in the posterior urethra,one died of metastasis tumor 18 months after operation,and the other died 32 month after operation.Conclusions Combined use of various kinds of the diagnostic means (ultrasound,IVU,R-P,CTU,MRU,cystoscopy,ureteroscopy) are important for the diagnosis of urothelial tumor in multiple organs.It needs to select the operate mode according to the tumor staging and grade and the patient's condition.Reinforcement surveillance and close follow up is required after operation.

Objective To develop a small-diameter tissue-engineered blood vessels which possesses normal blood vessels physiological structure, good biocompatibility, and mechanical properties. And it was evaluated by mechanical and biological of national standard of medi-cal transfusion material. Methods The bio-derived material were regarded as the ground substance, and it was evaluated by mechanical and biological of national standard after composite modification. Results The axial and radial tensile stress of the blood vessel was 23. 14 N and 36. 79 N respectively, and it was greater than the standard 7. 5N. The tensile rate of the axial and radial was 95. 19% and 80. 24% respec-tively, which were higher than the standard value 20%. The suture strength of the blood vessel was 13. 71 N, which was conform to the me-chanical requirement. Mainly used blood vessels or its extracts to detect the pH of the blood vessels is in the scope of control deionized water pH (7. 5 ± 1. 5);the hemolysis rate was 1. 3972% which was less than 5%;the whole blood coagulation time was 50% longer than the con-trol level, and there was no stimulation after intradermal injection. Conclusion With bio-derived material as the ground substance and com-positely modified, this kind od blood vessels is conform to the mechanical and biological of national standard, and it has the potential of clini-cal application which could play an important role in the replacement therapy of small-diameter vascular xenografts.

ObjectiveTo review the major complications in patients after transurethal electrovaporization of the prostate (TUVP) and transurethal plasmakinetic resection of the prostate (PKRP) retrospectively and to analyze the causes and management.MethodsClinical data of 92 cases of patients after TUVP and 226 cases after PKRP were reviewed retrospectively.The patients' relevant circumstances including subjective symptoms,objective indexes and the major long-term complications were followed up about 1-,3-,and 5-year after operation.Different therapeutic methods were chosen according to different causes of the complications.ResultsThere were no significant differences (P ＞ 0.05 ) between TUVP group and PKRP group in IPSS (7.3±2.8,7.2±2.5),QOL (2.6±0.7,2.7 ±0.5),Qmax[ (25.2±3.5),(25.5 ±3.8) ml/s] and PVR [(18.7 ±5.4),(17.8 ±6.3)ml].The incidences of bladder neck restriction was about 1.1％,3.3％,and 2.3％ after 1,3,and5 years in patients after TUVP,and 0.9％,2.7％,and 1.8％ after PKRP accordingly.For urethral stricture,it was about 3.3％,2.2％,and 1.1％ after TUVP,and 3.1％,2.2％,and 0.9％ after PKRP.For residual prostatic hyperplasia,it was about 1.1％,2.2％,and 4.5％ after TUVP,and 1.3％,2.7％,and 3.7％ after PKRP accordingly.ConclusionsTUVP and PKRP are effective and safe treatment options for BPH.The major long-term complications after TUVP and PKRP are bladder neck restriction,urethral stricture and residual prostatic hyperplasia.Regular and long-term follow-up is required for patients after TUVP and PKRP.Different therapeutic methods should be chosen according to different causes of the complications after operation.

This study was conducted to examine the effectiveness of a gene transfer of human TGF beta 1 gene into smooth muscle cells and whether the TGF beta 1 can increase elastin expression of smooth muscle cells. With the help of DOTAP, smooth muscle cells were transfected with pMAMneoTGF beta 1. The positive cell clones were selected with G418. The stable transfection and expression of TGF beta 1 in the smooth muscle cells were determined by immunofluorescence analysis. The expression of elastin in the transfected and untransfected cells were determined by in situ hybridization. The adhesion force between smooth muscle cells and matrix was detected by micropipette system. The results showed abundant TGF beta 1 stable expression in smooth muscle cells. TGF beta 1 gene can increase two-three times elastin expression and increase the adhesion between smooth muscle cells and matrix. TGF beta 1 can be used in vascular tissue engineering to increase smooth muscle cells adhesion.
Cell Adhesion ; Cells, Cultured ; Elastin ; biosynthesis ; Humans ; In Situ Hybridization ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics ; physiology ; Transforming Growth Factor beta1

To investigate the biomechanical behavior of human intestines. The tensile test human intestine was performed with the electronic tension machine in this paper. The results indicate that the exponential relationship for the stress-strain of the human intestine was obtained, and the exponential coefficient a of each segment of the intestine is almost the same although the constant C is different. It also shows that the relative rate of stretch length of each segment intestines is different in longitudinal and circumferential directions. And the incremental elastic modulus of colon is less than those of small intestine. It is considered that the colon can be more easily deformed. The experimental results provide the theoretic basis for research on intestinal endoscopic microrobot.
Biomechanical Phenomena ; Colon, Transverse ; physiology ; Elasticity ; Humans ; In Vitro Techniques ; Intestine, Small ; physiology ; Stress, Mechanical ; Tensile Strength

In this study, we prepared the acellular bone matrix of the inbred-line Banna mini-pig by using tissue engineering method and evaluated its possible application in bone tissue engineering. Histological analysis, xenoantigen expression and biomechanical measurement were performed on the matrix. HE staining and scanning electron microscopy showed the cellular components were almost removed. Immunohischemical result demonstrated that the xenoantigen, alpha-gal,was also eliminated. There was no statistically significant difference between the acellular bone matrix group and control group. The acellular bone matrix can provide appropriate space structure and strength for grafts. In conclusion, our data suggest that acellular bone matrix is a new kind of ideal bone scaffold material.
Animals ; Antigens, Heterophile ; analysis ; Biomechanical Phenomena ; Bone Matrix ; immunology ; Female ; Male ; Stress, Mechanical ; Swine ; Swine, Miniature ; Tissue Engineering ; alpha-Galactosidase ; analysis