Objective To evaluate the biocompatibility of vessel extracellular matrix (VECM) with bladder smooth muscle cells of rabbits,and to discuss the feasibility of vessel extracellular matrix as a matrix for urinary tract reconstruction.Methods Primary cuhured bladder smooth muscle cells (RBSMCs) iso- lated from New Zealand rabbits were implanted on VECM (1?10~6 cells/ml).The effect of VECM on meta- bolic activity,attachment,proliferation of RBSMCs were monitored in vitro by inverted light microscopy and scanning electron microscopy.The extracts of VECM and emulsion were prepared as experimental group and positive controls separately.The culture medium was used as negative control,and simple culture medium without cells was used as blank control.The cell viability was monitored by MTT method after 1-,3-,5-d see- ding.The in vivo tissue response to VECM was investigated by implanting into the subcutaneous sites of the rabbits.Results VECM exhibited nontoxic and bioactive effect on RBSMCs.RBSMCs could be attached to and proliferated on VECM and remained their morphologies.The cell proliferation rates of experimental group were 95.61%、98.34%、102.91%,respectively,after 1,3,5 d;those of negative control group were 100.00% ,respectively;and those of positive control group were 35.14%、38.95%、32.66%,respectively. There was significant difference in the rate between experimental group and positive control (P＜0.01),and no significant difference in the rate between experimental group and negative control (P＞0.05).In vivo, VECM demonstrated favorable tissue compatibility without tissue necrosis and fibrosis.Conclusions VECM exhibits nontoxic and bioactive effects on primary cultured bladder smooth muscle cells.It is a suit- able material for urinary tract reconstruction.

Gymnastics ; physiology ; Humans ; Workload

Advances in mechanism and application of the heating effect in breeding of microorganism are reviewed in this paper. Heat produces mutagenesis effect and screening effect. Heating mutagenesis effect is occurred through the substitution of G-C base pair induced by heat, and heating screening effect produces higher forward mutation rate induced by other mutagens.

Objective To study the level of serum lipids and body-fat content of high-fat diet induced pbesity rats(DIO).explore the relationship between intracellular caleium and ventrieufar arrhythmia.Methods Sixty male Sprague-Dawley(SD)rats were divided into control group(15)and experiment group(45),high-fat diet was administrated for 12 weeks to established obesity model,15 rats were selected into obesity group according to their body weight gain.The standard 2-lead electrocardiograph was used to detect the incidence and scores of arrhythmia induced by barium chloride(BaCl2,0.1 mg/kg)for 1 hour on every 8 rats from different groups respectively.Body-fat content.the level of serum total cholesterol(TC),triglyceride(TG),low density lipopmtein cholesterol (LDL-C) and high density lipopmtein cholesterol (HDL-C)were measured.The epididymal(EP),retroperitoneal (RE) and mesenteric(ME)white adipose pads was measured to obtain the body fat content.Single ventricular myrocytes of rats were isolated by enzymatic dissociation.The confocal laser scanning microscope was used to record basic intraceUular calcium level([Ca2+]i).Results The body-fat content in obesity group[(7.71±0.74)%] was significantly higher than control group[(4.69±0.37)%](t＝3.650,P＜0.05).The level of serum TC,TGand LDL-C were significantly higher(t＝3.801,2.778,3.536.P＜0.05) in obesity group[(1.26±0.04),(0.58±0.10),(0.51±0.04)mmol/L]than those in control group[(0.92±0.08),(0.29+0.03),(0.31±0.04) mmol/L].The level of serum HDL-C wa8 decreased gradually from control group[(0.53±0.05)mmol/L] toobesity group[(0.52±0.02)mmol/L],but there waft no significant difference between them(t＝0.186,P＞0.05).The incidence of arrhythmia induced by BaCl2(0.1 mg/kg)in obesity group was significantly higher than controlgroup(X2＝5.333,P＜0.05),and the scores of arrhythmla was increased in obesity group(2.5±0.6)too.The fluorescence intensity standing for[Ca2+]i was increased significantly(t＝2.409,P＜0.05)from obesity group(247.96±20.03)to control group (174.25±23.13).Conclusion As the free cytosolic calcium begin to accumulate,the arrhythmia morbidity is increased in obesity rats.

Australia ; Environmental Monitoring ; Humans ; Occupational Health ; Safety Management ; methods ; organization & administration

BACKGROUNDUrethral reconstruction for both congenital and acquired etiologies remains a challenge for most urologic surgeons. Tissue engineering has been proposed as a strategy for urethral reconstruction. The purpose of this study was to determine whether a naturally derived extracellular matrix substitute developed for urethral reconstruction would be suitable for urethral repair in an animal model.

METHODSA urethral segmental defect was created in 20 male rabbits. The urethral extracellular matrix, obtained and processed from rabbit urethral tissue, was trimmed and transplanted to repair the urethral defect. Then, the regenerated segment was studied histologically by haematoxylin-eosin staining and Van Gieson staining at 10 days, 3 weeks, 6 weeks, and 24 weeks postoperation. Retrograde urethrography was used to evaluate the function of the regenerated urethras of 4 rabbits 10 and 24 weeks after the operation. The urodynamics of 4 rabbits from the experimental group and control group I were assessed and compared. In addition, 4 experimental group rabbits were examined by a urethroscope 24 weeks after the operation.

RESULTSAt 10 days after operation, epithelial cells had migrated from each side, and small vessels were observed in the extracellular matrix. The matrix and adjacent areas of the host tissue were infiltrated with inflammatory cells. The epithelium covered the extracellular matrix fully at 3 weeks postoperation. Well-formed smooth-muscle cells were first confirmed after 6 weeks, at which point the inflammatory cells had disappeared. At 24 weeks postoperation, the regenerated tissue was equivalent to the normal urethra. Urethrography and urodynamic evaluations showed that there was no difference between normal tissue and regenerated tissue.

CONCLUSIONSUrethral extracellular matrix appears to be a useful material for urethral repair in rabbits. The matrix can be processed easily and has good characteristics for tissue handling and urethral function.

Animals ; Extracellular Matrix ; metabolism ; Rabbits ; Tissue Engineering ; methods ; Urethra ; pathology ; surgery

OBJECTIVETo investigate the curative effect and histocompatibility of reconstruction of traumatic urethral defect of rabbit using urethral extracellular matrix (ECM).

METHODSUrethral ECM was obtained by excision of the urethra in 20 donor rabbits. In experimental group, 20 rabbits were resected a 1.0 cm-1.5 cm segment of the urethra and artificially made a model of traumatic urethral defect, then reconstructed by the urethral extracellular matrix of the same length. The rabbit immunity response was assessed by lymphocyte transformation test and serum TNF-alpha level. The reconstructed urethral segments were stained with hematoxylin-eosin and Van Gieson stain and observed by histological examination postoperatively. The urethrography, urethroscopy and urodynamic examinations were performed.

RESULTSThere was no significant difference in stimulative index of lymphocyte transformation between ECM group and control group. The serum TNF-alpha levels of ECM group slightly rose, but the increase was not significant as compared with control group. On postoperative day 10, epithelial cell had migrated from each side and small vessels were found in the extracellular matrix. In the 3rd week, several layers of urothelium covered the whole surface of the matrix tube. In the 6th week, the disorganized arrangements of smooth muscle fibers were firstly observed by Van Gieson staining. In the 24th week, the smooth muscle cells increased and the matrix tube appeared fairly similar to normal urethral wall components. The urethroscopy and urodynamic evaluation revealed that the surface of reconstructed urethra was smooth and emiction was unobstructed.

CONCLUSIONThe urethral extracellular matrix might be an ideal and safe biomaterial for the reconstruction of urethral traumatic defect.

Animals ; Extracellular Matrix ; immunology ; physiology ; Female ; Immunohistochemistry ; Lymphocyte Activation ; Rabbits ; Reconstructive Surgical Procedures ; methods ; Tumor Necrosis Factor-alpha ; blood ; Urethra ; immunology ; injuries ; surgery

Adult ; Blood Donors ; China ; epidemiology ; Female ; Genotype ; HIV Infections ; complications ; epidemiology ; virology ; Hepacivirus ; genetics ; Hepatitis C ; complications ; epidemiology ; virology ; Humans ; Male ; Middle Aged

Objective To investigate the long-term follow-up results of patients treated with radical nephrectomy for small renal carcinoma.Methods Between January 1992 and June 2004,a total of 56 pa- tients(41 men and 15 women;mean age,54 years;age range,19-71 years)underwent radical nephrectomy for small renal carcinoma.The clinical data and long-term follow-up results of the 56 cases were analyzed ret- rospectively.All the patients were followed by questionnaire;and the 5-and 10-year survival rates were calcu- lated by Kaplan-Meier method.Results None of the patients was found to have metastasis preoperatively. Postoperative pathology showed renal clear cell carcinoma in 44 cases,granular cell carcinoma in 7,and mixed cell carcinoma in 5.Among these cases,ipsilateral adrenal metastasis was found in 1 case,local lymph- aden metastasis in 2,and perirenal fat metastasis in 4.Postoperatively,49 of the 56 patients(87.5%)were followed for 11-155 months with a mean of 71 months.The 5-and 10-year survival rates were 81.7% and 66.9%,respectively.Five patients died of metastasis from renal carcinoma.Conclusions Radical ne- phrectomy for small renal carcinoma has favorable long-term effects,therefore it is an optimal surgical proce- dure for small renal carcinoma.

Autophagosomes derived from tumor cells have been proved to induce potent T cell response both in mouse and human. In human in vitro study, dendritic cells(DC)loaded with cytomegalovirus(CMV)pp65 antigen-containing DRibble were capable to efficiently re-stimulate pp65-specific T-cell recall responses from freshly isolated or frozen humanperipheral blood mononuclear cell(PBMC). This study developed more robust assays using in vitro expanded antigen-specific T cells that contained a much higher percentage of antigen-specific T cells. DC cross-presentation efficiency of OX40 and CD80 modified pp65-DRibble was detected by intracellular IFN-γ staining. Compared with Ctrl/pp65 DRibble, the percentage of IFN-γ+ in total CD8+ T cells andCD4+ T cells was improvedwith OX40/pp65 DRibbleand CD80/pp65 DRibble stimulation. In addition, vaccine induced IL-12indendritic cells, whichpolarizes Th cells toward the IFN-γ high Th1 phenotype, evaluated by ELISA inco-culture supernatantwas dramatically higher in OX40/pp65 DRibble and CD80/pp65 DRibblegroups than in Ctrl/pp65 DRibble group. These results have implications for the immuneactivity of OX40 and CD80 modified DRibble and choice for prospective clinical use ofDRibble-based cancer immunotherapy.