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1.
Journal of Southern Medical University ; (12): 365-370, 2016.
Article in Chinese | WPRIM | ID: wpr-264039

ABSTRACT

<p><b>OBJECTIVE</b>To explore the feasibility of preparing ureteral acellular matrix (UAM) using perfusion systems.</p><p><b>METHODS</b>Using the luminal structure of the ureter, the UAM was prepared by perfusing canine ureter with SDS, TritonX-100, or both. The residual nuclei in the UAM were evaluated using HE staining, DAPI staining, DNA quantification, and agarose gel electrophoresis. The three-dimensional ultrastructure and the bioactive components were evaluated by Masson's trichrome staining, Alcian Blue staining, collagen quantification, GAG quantification, scanning electron microscopy (SEM), and toxicity detection.</p><p><b>RESULTS</b>HE staining and DAPI staining showed the absence of obvious nuclear materials in the combined group, which was further confirmed by DNA quantification and agarose gel electrophoresis. Masson's trichrome staining, Alcian Blue staining, collagen quantification and GAG quantification all verified that the ultrastructure and the bioactive components were well preserved in the combined group. SEM showed a large amount of porous structure on the surface of the UAM prepared by combined perfusion, and toxicity assay confirmed that the prepared UAM was nontoxic.</p><p><b>CONCLUSION</b>Perfusion of canine ureter with SDS and TritonX-100 is feasible to prepare UAM for ureteral reconstruction.</p>


Subject(s)
Animals , Dogs , Collagen , Metabolism , Extracellular Matrix , Microscopy, Electron, Scanning , Perfusion , Staining and Labeling , Tissue Engineering , Tissue Scaffolds , Ureter , Cell Biology
2.
Chinese Medical Journal ; (24): 928-932, 2015.
Article in English | WPRIM | ID: wpr-350376

ABSTRACT

<p><b>BACKGROUND</b>In order to improve the clinical treatment level of urinary system injury, it is necessary to build up an animal model of urinary system wound, which is not only analogous to real clinical practice, but also simple and practical.</p><p><b>METHODS</b>We have developed the third generation of firearm fragment wound generator based on the first and the second producer. The best explosive charge of the blank cartridge was selected by gradient powder loading experiments. The firearm fragment injuries were made to the bulbous urethra of 10 New Zealand male rabbits. One week preoperatively and 2, 4 and 8 weeks postoperatively, all the animals underwent urethroscopy and urethrography. At 2, 4 and 8 weeks postoperatively, two animals were randomly selected and killed, and the urethra was cut off for pathological examination.</p><p><b>RESULTS</b>The shooting distance of the third generation of firearm fragment wound generator is 2 cm. The best explosive charge of the blank cartridge is 1 g of nitrocotton. All rabbits survived the procedures and stayed alive until they were killed. Injuries were limited to bulbous urethra and distal urethra. Round damaged areas, 1-1.5 cm in length, on the ventral wall were observed. Ureteroscopy results showed that canal diameter gradually shrank by over 50% in 9 rabbits. The rate of success was 90%. Urethrography result noted that a 1-1.3 cm stricture was formed at the bulbous urethra. Histology results of injured stricture urethra showed that fibrous connective tissue hyperplasia and hyaline degeneration caused further stricture in the canal.</p><p><b>CONCLUSIONS</b>The third generation of firearm fragment wound generator imitates the bullet firing process and is more accurate and repeatable. The corresponding rabbit model of traumatic complex urethral stricture simulates the real complex clinical conditions. This animal model provides a standardized platform for clinical researches on treating traumatic injuries to the urinary system.</p>


Subject(s)
Animals , Male , Rabbits , Disease Models, Animal , Penis , General Surgery , Urethra , General Surgery , Urethral Stricture , General Surgery
3.
Chinese Medical Journal ; (24): 3087-3092, 2011.
Article in English | WPRIM | ID: wpr-319192

ABSTRACT

<p><b>BACKGROUND</b>Efficient cell adhesion and proliferation is a central issue in cell-based tissue engineering, which offers great promise for repair of urethral defects or strictures. This study evaluated the adhesion and growth of rabbit uroepithelium on a surface-modified three-dimensional poly-L-lactic acid (PLLA) scaffold.</p><p><b>METHODS</b>Urethral mucosa were harvested from male New Zealand rabbits and the urothelium were dissociated and then cultured. Immunocytochemistry on cultured uroepithelium for pancytokeratin and uroplakin II and TE-7 confirmed pure populations. After in vitro proliferation, cells were seeded onto a surface-modified urethral scaffold with non-knitted filaments. The morphology and viability of the cells were examined by immunohistochemical and fluorescence staining. Inverted and scanning microscopes were used to document cell growth and adhesion.</p><p><b>RESULTS</b>Three to five days after primary culture, the uroepithelial cells gradually became confluent, assuming a cobblestone pattern. The filaments of the urethral scaffold had excellent biocompatibility and allowed growth of the uroepithelium, without affecting viability. The uroepithelial cells adhered to and grew well on the scaffold. After 3 - 7 days, the cells grew vigorously and meshes of the scaffold were full of uroepitheliums.</p><p><b>CONCLUSIONS</b>The surface-modified urethral scaffold with non-knitted filaments allows the growth of uroepithelium and can serve as a carrier for the tissue engineering of urethra.</p>


Subject(s)
Animals , Male , Rabbits , Absorbable Implants , Cells, Cultured , Epithelial Cells , Physiology , Lactic Acid , Polyesters , Polymers , Tissue Engineering , Methods , Urethra , Cell Biology
4.
Chinese Journal of Gastrointestinal Surgery ; (12): 598-602, 2009.
Article in Chinese | WPRIM | ID: wpr-259358

ABSTRACT

<p><b>OBJECTIVE</b>To identify associated proteins involved in the molecular response of ischemic preconditioning (IPC) against intestinal ischemia/reperfusion (II/R) in the intestinal mucosa of rats.</p><p><b>METHODS</b>Sixteen SD rats were randomly divided into II/R and IPC groups. II/R injury in rats was produced by clamping superior mesenteric artery for 60 min followed by 60 min reperfusion. IPC was elicited by 20 min ischemia and 5 min reperfusion before index ischemia. The intestinal mucosa was scratched immediately after 60 min of reperfusion and total proteins were separated by immobilized pH gradient (IPG) based on two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed using Image Master 2D Elite 5.0 image analysis software and identified by MALDI-TOF-MS. The biological information of these proteins was searched in the database of these peptide mass finger-printing (PMF). Western blotting and RT-PCR were used to validate the differentially expressed proteins.</p><p><b>RESULTS</b>Image analysis revealed that averages of 1404+/-20 and 1338+/-20 were detected in II/R and IPC groups. A total of 10 spots yielded good spectra, and 8 spots matched with known proteins after database searching. These proteins were mainly involved in anti-oxidation, inhibiting apoptosis and energy metabolism. Western blot confirmed up-regulation of aldehyde dehydrogenase and RT-PCR confirmed up-regulation of aldose reductase in IPC group.</p><p><b>CONCLUSION</b>The clues provided by comparative proteome strategy will shed light on molecular mechanisms of IPC against II/R injury.</p>


Subject(s)
Animals , Male , Rats , Intestinal Diseases , Metabolism , Pathology , Intestinal Mucosa , Metabolism , Pathology , Ischemic Preconditioning , Proteomics , Rats, Sprague-Dawley , Reperfusion Injury , Metabolism , Pathology
5.
Chinese Journal of Hypertension ; (12)2007.
Article in Chinese | WPRIM | ID: wpr-685926

ABSTRACT

Objective To explore the underlying relationship between hyperglycemic factors in type 2 diabe- tes.Methods Fifty seven type 2 diabetes with obesity (DM-OB)and 64 without obesity(DM-NOB)were recruited. Age,body mass index(BMI),hemoglobin A1c (HbA1c),homeostasis model assessment-2 insulin resistance (HO- MA-IR),high-sensitivity C-reactive protein (hsCRP),fasting plasma glucose,postprandial plasma glucose (PPG), postprandial glucose excursion(PPGE),lipid profile,blood pressure were determined.Results DM-OB subjects had significantly higher HOMA-IR,BMI,DBP,TC,hsCRP,HbAlc,LDL-C when compared with DM-NOB sub- jects.Pearson correlation analysis,in DM-OB subjects,BMI,FBG,FPG,HOMA-IR,hs-CRP were all the posi- tive relative factors(P all

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