Purpose:To investigate the effect of replication defective retroviral vectors carried sense or antisense TGF?1 fragment on the cell cycle regulation and proliferation of human bladder cancer. Methods:The replication defective retroviral vectors that integrated sense or antisense bioactive fragment of transforming growth factor?1 were constructed,and named as pRevT? and pRevT?-AS respectively. The influence of each vector on the cell proliferation,clone-formation and alteration of cell cycle of bladder cancer cell line EJ were observed in vitro.Results:The titre of pRevT? and pRevT?-AS were 0.84,0.88?10 5 CFU/ml respectively,the vectors integrated to EJ cells and expressed efficiently. Inhibition TGF?1 gene expression reduced proliferation and clone-formation rates of EJ cells. The G 0 /G 1 stage ratios in the antisense TGF?1-transfected EJ cells were increased,simultaneously,the S stage ratios were decreased. Conclusions:The antisense TGF?1 vector can reduce the expression of endogenous TGF?1 in EJ cells,induce G 1 stage arrest and inhibit proliferative growth in vitro.

Objective To discuss the clinical value of three dimensional proton MR spectroscopic imaging( 3D 1HMRSI) in the detection of transition zone(TZ) prorate cancer and evaluate the feasibility of 3D 1HMRSI for determining the aggressiveness of TZ cancer by analyzing its metabolic characteristics.Methods The 3D 1HMRSI data of sixty patients suspected TZ cancer in conventional MR examinations were retrospectively analyzed.The values of (Cho + Cre)/Cit of TZ cancer and benign prostatic hyperplasia (BPH) voxels were recorded and compared using independent sample t' test,and the area under the ROC curve was used to evaluate the diagnostic accuracy.Based on Gleason scores,TZ cancer voxels were divided into three groups,including low-risk ( Gleason score ＜ 7 ),intermediate-risk ( Gleason score =7 ) and highrisk (Gleason score ＞ 7).The values of (Cho + Cre)/Cit were compared among the three groups using Kruskal-Wallis test.The correlation of the value of (Cho + Cre)/Cit and Gleason score was analyzed using rank correlation analysis.Results Among the 60 patients,histopathology confirmed TZ cancer in 25 patients and BPH in 35 patients.The inversion of Cho and Cit peak value with increased ( Cho + Cre)/Cit was detected in 160 out of 177 TZ cancer voxels.Most spectral curves of the 517 BPH voxels were similar with that of normal peripheral zone on 1HMRSI.The mean values of ( Cho + Cre)/Cit of TZ cancer and BPH voxels were 2.17 ± 1.29 and 0.77 ± 0.20,respectively,with significant difference between them( t' =14.38,P ＜ 0.01).Using (Cho + Cre)/Cit for distinguishing TZ cancer,the area under ROC curve was 0.985 (P ＜ 0.01).With the cut-off point 1.08,the sensitivity,specificity and accuracy of TZ cancer diagnosis was 92.7％,94.2％ and 93.8％,respectively.The number of low-risk,intermediate-risk and high-risk TZ cancer voxels were 57,64 and 56 respectively,and the mean values of ( Cho + Cre)/Cit of the three groups were 1.43 ( 1.16-1.87 ),1.66 ( 1.43-2.36 ) and 2.32 ( 1.86-3.30 ) respectively,with significant difference among them ( x2 =36.282,P ＜ 0.01).The value of ( Cho + Cre )/Cit of TZ cancer increased with the increasing of Gleason score with positive correlation ( r =0.555,P ＜ 0.01).Conclusions TZ caner has a different metabolic profile from BPH.The value of ( Cho + Cre)/Cit on 3D 1HMRSI could provide useful information for early detection and differentiation TZ cancer from BPH.Besides these,( Cho +Cre)/Cit is also a valuable imaging marker to evaluate the aggressiveness of TZ cancer.

Objective To evaluate the real-time genotyping and quantitative PCR(RT-GQ-PCR)method by comparing it with direct sequence analysis and the multiplex-PCR method.Methods RT-GQ-PCR,direct sequence analysis and the multiplex-PCR method were used to detect HBV genotypes of 113 patient samples with HBV-DNA positive.ResultsThe detection rate of RT-GQ-PCR and direct sequence analysis was 100%,and the multiplex-PCR is 94.69%.The concordance between RT-GQ-PCR and the multiplex-PCR is perfect(Kappa value =0.915),and the consistency of RT-GQ-PCR and direct sequence analysis is pretty good(Kappa value = 0.742),specially at detecting single genotype.Twenty-eight samples with genotypes B and C dual infections were detected by RT-GQ-PCR,but only 19 samples by the multiplexPCR and 13 samples by direct sequence analysis.Conclusion The RT-GQ-PCR is convenient,rapid and accurate in HBV genotyping,especially more sensitive than direct sequence analysis and the multiplex-PCR for detecting dual genotypes.The method is applicable for large-scale epidemiological study.

Objective to explore the role of Beclin 1 in gastric carcinogenesis and subsequent progression. Methods MkN28 cells were trans-fected with Beclin 1-expressing plasmid,and then the proliferation and cell cycle was measured by CCk-8 and PI staining. Beclin 1 expression was examined by immunohistochemistry and in situ hybridization on tissue microarrays containing gastric cancers,adjacent non-neoplastic mucosa,and metastatic lymph node. the correlation with the tumorgenesis,clinicopathological and prognostic parameters was analyzed. Results Beclin 1 overex-pression resulted in G2 arrest of MkN28 cells and reduced the proliferation. Beclin 1 mRNA was highly expressed in gastric cancer than matched mu-cosa by ISH(P < 0.05). Beclin 1 was highly expressed in male than female patients with gastric cancer(P < 0.05). the elder patients with gastric cancer had higher Beclin 1 expression than younger ones(P < 0.05). the diffuse-type carcinomas showed less Beclin 1 expression than intestinal and mixed type ones(P < 0.05). kaplan-Meier analysis indicated that Beclin 1 expression was positively correlated to favorable prognosis of the can-cer patients(P < 0.05). Conclusion Beclin 1 expression is closely linked to pathogenesis,metastasis and differentiation of gastric cancer. Beclin 1 might be employed to indicate the favorable prognosis of gastric cancer patients and regarded as a target of gene therapy.

Objective To clarify the clinicopathological significance and the reversing effects of BTG3 expression on the aggressive phenotype in gastric cancer. Methods BTG3 expression was detected in gastric cancer tissues by on tissue microarray and immunostaining. BTG3?expressing plasmid was transfected into MKN28 and MGC803 cells,the proliferation,cell cycle,differentiation and autophagy were analyzed by CCK?8,PI staining,alkaline phosphatase activity and GFP?LC?3B transfection,respectively. Results BTG3 overexpression inhibited cell proliferation,in?duced S/G2 arrest,differentiation and autophagy in both cells(P<0.05). BTG3 expression was decreased in gastric cancer in comparison with the adjacent mucosa(P<0.05),and positively correlated with venous invasion and dedifferentiation of the cancers(P<0.05). Conclusion BTG3 ex?pression contributes to gastric carcinogenesis and subsequent progression. BTG3 overexpression can reverse the aggressive phenotypes,which could be employed as a potential target for gene therapy of gastric cancer.

Objective To investigate the value and diagnostic efficiency of the quantitative dynamic contrast enhanced-magnetic resonance imaging (DCE-MRI) and intravoxel incoherent motion (IVIM) parameters using three dimention (3D)-histogram analysis for discriminating the Gleason score (GS) of prostate cancer. Methods A total of 53 patients pathologically confirmed as prostate cancer by systemic prostate biopsy who had routine , DCE and DWI-MRI scans were retrospectively analyzed. There were 15 cases for low-risk and 38 cases for intermediate/high-risk prostate cancer. The 3D ROI of all lesions based on T2WI was achieved by image registration to get the quantitative parameters of DCE-MRI and DWI-IVIM. The parameters of DCE-MRI contains: transfer constant (Ktrans), rate constant (Kep) and extracellular-extravascular volume fraction (Ve).The DWI-IVIM related quantitative parameters were ADC, diffusion coefficient (D), diffusion coefficient related to perfusion (D*) and perfusion fraction (f). Then the histogram analysis of these quantitative parameters was performed to get the mean, median, 25th percentile, 75th percentile, Skewness and Kurtosis. Using the Spearman rank correlation analysis to evaluate the correlation of these parameters and GS of prostate cancer. The diagnostic performance of these quantitative histogram parameters related to the GS in identifying low-risk and intermediate/high-risk of prostate cancer was carried by ROC. Results The Kep and Ktrans (mean, median, 25th, 75th) of DCE-MRI were positively correlated with GS (r value was 0.346 to 0.696, P<0.05). The ADC (mean, median, 25th, 75th), D (mean, median, 25th, 75th, Skewness, Kurtosis) and D*(25th) of DWI-IVIM were correlated with GS (r value was-0.544 to 0.428, P<0.05). The DCE-MRI quantitative parameters Kep (25th) had the highest area under curve (AUC, 0.961); The ADC (median) and D (25th) had higher AUC( 0.832, 0.888) in the quantitative parameters of DWI-IVIM, the difference between Kep(25th) and ADC (median) was statistically significant (Z value was 2.212, P value was 0.027). The difference of AUC between Kep (25th) and D (25th), D (25th) and ADC (median) was not statistically significant (Z values were 1.027 and 1.398, P values were 0.162 and 0.304, respectively).Conclusion DCE and IVIM quantitative parameters (Kep, Ktrans, ADC, D) histogram analysis results are correlated with GS, and can be used for distinguishing low-risk from intermediate/high-risk prostate cancer.

Objective To evaluate the diagnostic value of prostate imaging reporting and data system version 1 (PI-RADS V1) and version 2 (PI-RADS V2) for detection of prostate cancer (PCa) in the transition zone (TZ).Methods Seventy-seven patients with suspicious lesions in TZ on mpMRI were scored according to the PI-RADS system (V1 and V2) before MR-TRUS fusion guided biopsy prospectively.In all of the patients with suspicious tumors,respectively at least one lesion with a PI-RADS V1 assessment category of ≥3,was selected for biopsy.Independent sample t test was used to compare scores of PI-RADS V1 and V2 between PCa and benign prostatic hyperplasia (BPH).The diagnostic performance of PI-RADS V 1 and V2 for detection of PCa in the transition zone was compared by analyzing ROC basing on the results of MR-TRUS fusion guided biopsy.Results A cohort of 77 patients was performed including 31 cases of PCa (32 cores) and 46 cases of BPH (51 cores).PCa (V1:1 1.50±2.79;V2:4.28±0.99) had significantly higher scores of both PI-RADS V1 and PI-RADS V2 than BPH(V1:7.51± 1.63;V2∶2.61 ±0.67) (P＜0.05).Using a PI-RADS V1 score cut-off ≥ 11,sensitivity and specificity in group PCa and BPH were calculated,which were 68.8％(22/32) and 96.1％(49/51) with a area under curve of 0.869;using a PI-RADS V2 score cut-off ≥4,which were 75.0％ (24/32) and 90.2％ (46/51) with a area under curve of 0.888,respectively.Conclusions PI-RADS system can indicate the likelihood of PCa of suspicious lesions in TZ on Mp-MRI.PI-RADS V2 perform better than V 1 for the assessment of prostate cancer in TZ.

Objective To analyze the multimodal imaging findings of nonspecific granulomatous prostatitis (NSGP)and improve our understanding of the disease.Methods 1 427 patients underwent transrectal ultrasoundGguided prostate biopsies in our department,then 1 3 nodules in 11 patients were proven NSGP by pathology,of which 3 nodules were in inner gland and 10 were in outer gland.The characteristics of these nodules on MRI,transrectal ultrasound elastography (TRE)and contrastGenhanced ultrasonography (CEUS)were retrospectively analyzed. Results MRI:All of the nodules showed similar performances:hypointensity on T1 WI and T2 WI,low ADC and high signal on DWI (highGbGvalue),early enhancement and faded fast or slow,depressed citrate (Cit)and relatively elevated choline (Cho)peak.TRE:The cutGoff point of strain ratio (SR)was determined as 5.97 to diagnose prostate cancer and the SRs of 8 nodules were greater than 5.9 7.CEUS:1 1 nodules showed early enhancement and great intensity,but the rest of nodules showed same performance with normal tissue.Conclusion The multimodal imaging helps to improve the understanding of NSGP and plays a positive role in guiding targeted biopsy.Moreover,the operators could lessen the number of needle punctures properly according to the multimodal imaging findings, which would reduce the risk of puncture complications on patients.

OBJECTIVETo generate a mouse model of chronic hepatitis B (CHB) infection by performing in vivo transduction of hepatitis B virus (HBV) covalently closed circular (ccc)DNA.

METHODSNude mice were injected with HBV cccDNA at doses of 1.5, 1.0 or 0.5 mug/ml. A control group was generated by giving equal injection volumes of physiological saline. The serum levels of hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) on post-injection days 1 and 3, weeks 1-6, 8 and 10 were assayed by reflection immunoassay. At post-injection week 10, all animals were sacrificed and liver tissues were collected. Copies of HBV DNA in serum and liver tissue were detected by real-time PCR. HBV antigens in liver tissue were detected of by immunohistochemistry. Pathological analysis of liver tissue carried out with hematoxylin-eosin staining. Linear correlation of data was determined by statistical analysis.

RESULTSHBsAg and HBeAg were detected in sera from all three groups of cccDNA-injected mice staring at post-injection day 1 and lasting through week 10. The levels of HBsAg over the 10-week period showed two patterns of increase-decrease;the lowest level was detected at week 4 and the highest level was detected at week 8. In contrast, the levels of HBeAg over the 10-week period showed three patterns of increase-decrease; the lower levels were detected at weeks 2 and 4 and the higher levels at weeks 3 and 6. HBV DNA copies in liver tissues showed a cccDNA dose-dependent descending trend over the 10-week study period (1.5 mug/ml:1.14E+07 ± 6.51E+06 copies/g, 1.0 mug/ml:9.81E+06 ± 9.32E+06 copies/g, and 0.5 mug/ml:3.72E+06 ± 2.35E+06 copies/g; Pearson's r =0.979). HBV DNA copies in sera showed the pattern of 1.0 mug/ml cccDNA more than 1.5 mug/ml cccDNA more than 0.5 mug/ml cccDNA, and in general were higher than those detected in the liver tissues. Liver tissues from all cccDNA-injected mice showed positive immunohistochemistry staining for both HBsAg and HBeAg. HE staining showed that the liver tissues of all cccDNA-injected mice had severe fatty and vacuolar degeneration and less obvious structure of liver lobules (compared to the liver tissues from control mice).

CONCLUSIONThe CHB mouse model successfully established in this study by in vivo transduction of HBV cccDNA may represent a useful tool to study the pathogenic mechanisms and potential antiviral treatments of human CHB.

Animals ; DNA, Circular ; administration & dosage ; DNA, Viral ; administration & dosage ; Disease Models, Animal ; Hepatitis B Surface Antigens ; blood ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; physiology ; Hepatitis B, Chronic ; virology ; Male ; Mice ; Mice, Nude ; Transduction, Genetic ; Virus Replication

Objective To compare the effect and safety of stenting versus directional atherectomy (DA) in the treatment of TASCⅡ A and B superficial femoral artery lesions.Methods 100 patients with TASC Ⅱ A and B lesions were divided into percutaneous transluminal stenting(PTS) group (n =50) and DA group (n =50).Patients were compared in terms of technical success rate,treatment success rate,first operation cost,postoperative ankle brachial index (ABI),limb salvage rate,survival,and patency.Results The technical success rate in both PTS and DA group was 100％.The treatment success rate was 98％ vs.86％,P＞0.05.Postoperative ABI:0.82 ±0.19 vs.0.80 ±0.27,P＞0.05.First operation cost:(34 820 ± 1 051) yuan vs.(45 635 ± 1 358) yuan,P ＜0.001;All patients were followed-up for up to 2-year,the cumulative patency rate was 81.6％ vs.72.9％ (P＞0.05).Limb salvage rate was 97.9％ vs.93.8 ％,P ＞ 0.05.Conclusion There were no significant differences in the effect and safety of PTS versus DA in the treatment of TASCⅡ A and B superficial femoral artery lesions.