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

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

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.

AIM:To sieve the optimal purification of the total flavonoid from Mulberry Leaves by exploring 8 types of macroporous resin's static and dynamic adsoption-desorption.METHODS:The factors affecting separation,such as flow rate concentration,pH and kind of eluting agent were considerated.RESULTS:Experiment results showed that AB-8 resin possesses high absorption capacity.The best craft was:the adsorption and desorption-power was 2 BV/h,the content of the total flavonoid in liquid was 0.613 mg per 1 mL,pH 4,as the desorption solvent,the concentration of ethanol was 70%,regenerate the resin after using three times.CONCLUSION:Flavones content used the method can be up 2-fold.

Circulating tumor cells (CTCs) detections have unique advantages in the real-time personalized medicine of patients with tumors.The rarity and heterogeneity of CTCs in peripheral blood pose great technical challenge for CTCs researches.Most of technologies for detecting CTCs have been developed to enrich CTCs by virtue of physical properties or specific biological features of the cells at first,then genotype or phenotype analysis are performed to count or characterize CTCs.Advances in molecular analysis of single cells enable CTCs detections to provide more accurate and comprehensive information.Biological processes during CTCs invasion and metastasis such as epithelial-mesenchymal transition and clustering should to be considered seriously to promote the clinical applications of CTCs detection.

Object To study the feasibility of flocculation procedure on Bushenhuoxue Decoction, using chitosan instead of alcohol. Methods Using chitosan clarifier to the flocculation procedure of Bushenhuoxue Decoction; selecting the optimum content of chitosan and the density of decoction according to the total extract, and the content of icariin, comparing with the flocculation procedure by alcohol sedimentation. Results Chitosan clarifier could make the decoction clarify as alcohol does. On the retaining of active compounds, the former was better than the later. Conclusion Chitosan can be used to the flocculation procedure of Bushenhuoxue Decoction instead of alcohol.

Objective To probe the precision of the magnetic resonance spectroscopy (MRS)in assessing the fat content in the vertebral bone marrow and the changes at different vertebrae.Methods A total of 46 female volunteers were recruited in this study. After routine MRI examination,single voxel (SV)was chosen for the 1 H-MRS acquisition at L1 -L4 vertebral body.Thirty subjects were scanned twice to evaluate the precision of MRS.The fat fraction (FF%),unsaturation index (UI)and coefficients of variation (CV) were calculated by amplitude and area under the peak respectively.The changes of fat content in marrow at L1 -L4 vertebrae was further evaluated.Results The CV related to peak amplitude was 1.7% in quantification of vertebral FF%,while the CV related to peak area was 6.3%.In quantification of vertebral UI,the CV related to amplitude and area was 2.4% and 7.8% respectively.The FF% was ascended from L1 to L4,while UI of bone marrow declined.A significant correlation was observed between FF% and UI (r=-0.714,P <0.01).The correlation of FF% among L1 -L4 were also significantly observed (r =0.612 -0.923,P <0.01). Conclusion MRS at 3.0 T provides reliable measurement for marrow fat content with a good precision associated with amplitude. Meanwhile,there is a preferential increase in saturated lipids compared to unsaturated lipids from L1 to L4.There is a high correlation of FF%among L1-L4.MRS is a valuable tool for providing complementary information on osteoporosis research.

To find a component with the strongest anti-endotoxin effect from the four components (F021, F022, F023, and F024) of chloroform extract (F02) of Radix Isatidis, the protective effects of the five components on endotoxin-challenged mice and their effects on the production of TNFalpha and IL-6 by macrophages from the endotoxin-challenged mice were compared. It was found that the five components all had protective effects on endotoxin-challenged mice in terms of mortality. The mortalities of the mice challenged by 2.42 mg/kg endotoxin were 30 %, 50 %, 20 %, 50 % and 60 % after treatment with F02, F021, F022, F023, and F024. The 5 components all had inhibitory effects on the production of TNFalpha and IL-6 by macrophages from the mice. At a concentration of LPS of 50 ng/mL, the inhibitory rates of F02 , F021 1, F022, F023, and F024 for the production of TNFalpha and IL-6 were 76.54 %, 30.86 %, 78.60 %, 36.63 %, 2.06 % and 75.85 %, 18.45 %, 77.68 %, 24.41 %, 3.64% and at the dose of 100 ng/mL, the inhibitory rates were 49.65 Y, 16.86 %, 66.97 %, 13.39 %, 7.16 % and 59.78 %, 1.28 %, 61.86 %, 2.08 %, 1.44 %, respectively. It is concluded that the component F022 of Radix Isatidis has the strongest anti-endotoxin effect.

Objective This study investigates the relationship of serum irfflammatory cytokine levels with intracranial pressure (ICP) in patients with severe traumatic brain injury (TBI) after surgery.Methods A total of 32 cases with severe TBI and placement of ICP monitor were prospectively enrolled.Serum was collected before surgery and every 12 h after surgery.Cytokines levels of interleukin (IL)-1β,IL-8,and tumor necrosis factor (TNF)-α were analyzed and compared with outcome of patients.Hourly values of ICP were recorded.The degree of ICP above treatment threshold (20 mm Hg) were calculated every 12 h as pressure times time dose(PTD-ICP20),which was compared with serum cytokine levels before (Pre) and after (Post) the 12-hour time period using linear regression method.Results Serum IL-1β (P＜0.05),IL-8 (P＜0.01) and TNF-a (P＜0.01) levels elevated dramatically after severe TBI and were significantly associated with outcome of patients.Mean PTD-ICP20 was (42.9 ± 60.2)mm Hg/h and was correlated with increased Pre-IL-8 (r=0.554,P＜0.001),Pre-TNF-α (r=0.597,P＜0.001),Post-IL-8 (r=0.629,P＜0.001) and Post-TNF-α (r=0.538,P＜0.001) levels.Conclusion Serum IL-8 and TNF-α demonstrated the most promising candidate bio.rnarkers of impending ICP elevation in this study.These findings indicate a feasible way of monitoring patients with severe TBI.

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.