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@#ObjectiveTo observe the expression of vascular endothlial growth factor(VEGF) and throbospondin-1(TSP-1) in different grade bladder cancer.MethodsSpecimens of 70 cases of bladder transitional cell cancer including Grade Ⅰ 28 cases,Grade Ⅱ 27 cases,Grade Ⅲ 15 cases(according to WHO pathological grade) were stained with immunohistochemistry method.The expressing values of VEGF and TSP-1 were calculated by mean vascular density(MVD).All data was analyzed with SPSS 10.0,and with expression of VEGF and TSP-1 was detected with immunohistochemisrty double staining.Finally,the result of expression was observed by laser scan with focus microscope.ResultsThere was a positive correlation between the cancer grade and the expression of VEGF,and the expression of Grade Ⅰ was significant difference with that of Grade Ⅱ(P<0.01),but there was no difference between Grade Ⅱ and Grade Ⅲ(P>0.1).Expression of TSP-1 decreased with grade increasing but not showing a negative correlation,while,Grade Ⅰ was significant difference with that of Grade Ⅱ and Grade Ⅲ(P<0.01),Grade Ⅱ was not differcnt with Grade Ⅲ(P>0.1).In 28 Grade Ⅰ cases,values of MVD of VEGF and TSP-1 had no correlation(rs)=0.167,P>0.1).Double immunostaining detecting with laser scan with focus microcope indicate showed that VEGF and TSP-1 appeared in same bladder cancer specimen.ConclusionExpression of VEGF is gradually increased from Grade Ⅰ to Grade Ⅲ,and VEGF is a promoting factor of angiogenesic bladder cancer.Expression of TSP-1 is the strongest in Grade Ⅰ cases,and TSP-1 is an inhibiting angiogenesic factor,which inhibiting function only in early stage.Immunofluorescent staining can only provide evidence of together expression of VEGF and TSP-1,cannot evaluate the degree of expression.
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Objective To explore the mechanism of imperfect recovery of the renal function after release of chronic ureteral obstruction. Methods Adult New Zealand rabbits(n=36) were randomly divided into normal control group(n=3),obstructive control group(n=3) and experimental group(n=30).The latter two groups were subjected to chronic unilateral ureteral obstruction.Two weeks later,3 rabbits were sacrificed and the others underwent ureterostomy,which were sacrificed separately following 12 hours,24 hours,3 days,7 days,14 days.The renal papilla and cortex were examined by scanning electron microscopy. Results Shortly after the release of ureteral obstruction,massive loss of brush border and ballooning of epithelial cells were noted in proximal tubules.Distal tubules and collecting tubules also showed severe ballooning of epithelial cells and normal struction were not seen.Two weeks later,many tubular components became patent with normal epithelial lining.Some of them,however,were deteriorated further in spite of the release of obstruction.Thus there was a marked tubular heterogeneity in recovery.Conclusion Further deterioration of some tubules might be one of the important pathologic basis for imperfect recovery of renal function after the release of chronic ureteral obstruction.
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Objective To study the ultrastructural changes in three different zones of benign prostatic hyperplasia(BPH). Methods The ultrastructural changes in peripheral,transitional and central zones(PZ,TZ and CZ)of prostates from 10 patients with BPH were observed with transmission electron microscopy. Results Glandular epithelia layers in TZ of BPH were increased.The stroma in TZ was mainly composed of smooth muscle cells (SMC) of vigorous proliferation and fibroblasts.Mast cell and lymphocyte with cytoplasma membrane merge were observed in this zone.In PZ and CZ of BPH, SMC showing degenerative changes,such as,nuclear pyknosis,dilatation of perinuclear space and vacuolation of cytoplasma were frequently seen. Conclusions There are significant differences in the ultrastructure of three zones in BPH.
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Objective To study the effect of tamoxifen on the proliferation and apoptosis of germ cells and to evaluate its clinical application to idiopathic oligospermia. Methods 41 oligospermia patients were treated with tamoxifen.Testicular tissues obtained by Tur Cut biopsy needle,both before and after the treatment,were observed under electron microscopy,the incidence of apoptosis and PCNA expression in germ cells being detected. Results Testicular pathological changes in different degree were observed on electron microscopy.Compared with the normal controls.PCNA proliferation index (PI) was lower ( P
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Objective To explore the effects of ureteropelvic junction(UPJ) obstruction on contralateral kidney.Methods Twenty-four adult New Zealand rabbits were divided randomly into 3 groups,ie,complete obstruction group(group 1,n=8),partial obstruction group(group 2,n=8) and shamoperation group(group 3,n=8).In groups 1 and 2, the right ureter was ligated completely or partially.In group 3,the right ureter was only exposed but not ligated.The animals were killed 4 weeks after operation,and the contralateral kidneys were resected.Before killing,anterior urography showed that right hydronephrosis occurred in all the rabbits in groups 1 and 2.The contralateral renal weight was measured by electron balance.The apoptosis in the contralateral renal cortex and medulla was detected by FITC-AnnexinV/PI flow cytometric assay.The morphological changes of the samples were also studied.Results After 4 weeks,the contralateral renal weight of the rabbits in groups 1,2 and 3 was(3.96?0.20),(3.47?0.19) and(3.20?0.21)g/kgBW,respectively.The difference of contralateral renal weight between groups 1,2 and group 3 was significant(P0.05).In contrast,the apoptosis rates of contralateral renal medulla cells in groups 1,2 and 3 were(23.75?2.34)%,(11.82?1.40)% and(2.36?0.65)%,respectively.There were significant differences between these 3 groups(P
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Objective To investigate the selective killing effect of adenoviral mediated herpes simplex virus-thymidine kinase/ganciclovir (HSV-tk/GCV) suicide gene system controlled by human telomerase reverse transcriptase (hTERT) promoter on bladder cancer cells in vitro. Methods Bladder cancer cell line 253J and human fibroblast cell line MRC-5 were transfected by the recombinant adenovirus of different multiplicities of infection (MOI),and the infection rate was measured by observing the expression of enhanced green fluorescent protein (EGFP) under the fluorescent microscopy.Ad-hTERT-HSV/tk and Ad-CMV-HSV/tk were transduced into 253J and MRC-5,followed by GCV treatment. The relative survival rate of cells in presence of prodrug GCV was measured with MTT method. Results Recombinant adenovirus Ad-hTERT-EGFP could selectively infect 253J cells,with the infection rate associated with the increasing MOI of recombinant adenovirus (P
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The changes of the morphology and percentage of the macroglia surrounding focus of brain stabbing injury (BSI) were observed by immunohistochemistry,immunoinfluorescence stain and flow cytometer(FCM) and the effect of magroglia during glial scar forming were elucidated.The results showed that a large number of GFAP-immunoreactive positive cells were accumulated around the focus of BSI.These cells were hyperplastic,hypertrophic,and emerged swollen cytoplasmic processes.The most marked changes were observed at 1-2 week after BSI.The results of FCM showed that the percentage of GFAP positive cells increased gradually and reached to a peak during 1～2 week after BSI.The peak ratio of GFAP positive was about 46%.However,the changes of morphology and number of GC positive cells were not detected after BSI.The authors believed that astrocyte is the main macroglia during glial scar formatting .The oligodendrocytes is not an active cell during this course.
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Objective To study the occurence, development and regulation of reactive gliosis with astrocyte (Ast) in vitro. Methods Ast was isolated and cultured in vitro and its model of reactive gliosis was established by scratching the cultured astrocytes. The reactivity and rules of Ast to injury was studied by morphological changes, RT-PCR, immunocytochemistry, in situ hybridization and imaging analysis. Results After scratching, the astrocytes showed typical features of reactive gliosis, with the hypertrophic cell body, thickened and lengtheded processes, and enhanced glial fibrillary acidic protein (GFAP) staining. In situ hybridization and RT-PCR analysis confirmed that the expression of GFAP mRNA was markedly increased. These changes occurred 1 d after scratching and reached the peak 5 to 7 d after injuring. Conclusion A model of reactive astrogliosis was successfully established in vitro which showed an active reaction to injury. The characteristics of reactive gliosis parallel that seen in vivo.
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Objective To investigate the therapeutic effects of traditional Chinese medicinal herbs(TCMH) on benign prostate hyperplasia(BPH) and their influence on proliferation and apoptosis of prostate cells in experimental mice. Methods A total of 120 male Kunming mice(weight,30 to 40 g)were randomly divided into 5 groups:①normal control group;②hyperplastic group;③normal saline(NS)(negative control)group;④TCMH group;⑤finasteride (positive control) group.Each group included 24 mice.The mice in normal control group were killed and their prostates were weighed.The BPH model was induced by implanting the urogenital sinus in the mice in other 4 groups.In the 61st day,the prostate weight of the mice in hyperplastic group was examined after they were killed;and the mice in the remaining groups were fed with NS,TCMH and finasteride,respectively.In the 91st day,the mice in these 3 groups were killed and their prostates were weighed.The proliferation and apoptosis of the prostate cells of mice in the 3 groups were determined by flow cytometry(FCM). Results The prostates in hyperplastic group [(149.30?8.46)mg] were heavier than those in TCMH group [(85.60?17.97)mg] ( P 0.05). Conclusions The TCMH has a significant therapeutic effects on experimental mice with BPH.The mechanisms may be as follows:TCMH can regulate the relevant genes of the prostate cells so that they can promote the apoptosis of the prostate cells and as a result,the volume and weight of the prostate of the mice are reduced.
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Objective To investigate the biocompatilility of human acellular amniotic membrane(HAM) with vascular smooth muscle cells(VSMCs) and to explore the possibility to construct tissue engineering bladder with HAM as the scaffold and VSMCs as the seed cells. Methods After physical and 1% trypsogen preparation,the HAM was mixed with VSMCs taken from rats for culture in vitro.Histological obserbation was done under inverted microscope and scanning electron microscope respectively.20 rats were divided into two groups.Hemicystectomies were performed in 20 rats,and 10 of them were repaired with HAM grafts with VSMCs on the half bladder,the other 10 were repaired with HAM grafts without VSMCs as the control group.The rats underwent postoperative assessment of bladder volume at the 2nd,4th and 8th weeks,and the grafts were observed by light microscope at the 2nd,4th and 8th weeks after surgery. Results The physical and 1% trypsogen treated HAM was pure with hollows and undamaged collagen fibers.The VSMCs could grow,adhere to and differentiate on the surface of HAM and into the hollows.At the 2nd,4th and 8th weeks after surgery, the bladder volumes of the experimental group were not different significantly compared with those of the control group.Epithelialization and smooth muscle cells regeneration occurred with the infiltration of inflammatory cells in the 2nd week after grafting,and the HAM were absorbed.In the 4th and 8th weeks,it was difficult to delineate the junction between the host bladder and grafts by histology.Conclusion HAM can be used as the scaffold to construct tissue engineering bladder as it has good biocompatibility with VSMCs without disturbing the cell form and the graft can be absorbed quickly.