Objective To investigate the cytotoxic effects and mechanism of PNP-CD chimeric gene vector originated from PNP/MeP-dR system on HCC cells. Methods The fusion suicide gene PNP-CD obtained by site directed mutagenesis technique was subcloned into pcDNA3.0 to construct a eukaryotic expression vector containing a chimeric gene, pcDNA3.0/ PNP-CD. After being identified by recombinant enzyme, PCR and subsequent sequencing, it was transfected into HepG2 cells by liposome-mediation method. The G418-resistant cellular clone with stable transfection of pcDNA3.0/PNP-CD, HepG2/PNP-CD was established by selection. The expression of PNP-CD gene was also verified by RT-PCR and Western blotting. The curve of cellular growth was assayed by Trypan blue exclusion. The cellular sensitivity of HepG2/PNP-CD to its specific prodrugs and its bystander effects were also assayed by MTT method. Results The chimeric gene, PNP-CD, was inserted into pcDNA3.0 correctly, and the stable expression of pcDNA3.0/PNP-CD in HepG2 cells was confirmed.This cellular clone was highly sensitive to its corresponding prodrugs. It was indicated that its bystander effects with the synergetic treatment of its specific prodrugs were substantially higher than those caused by the same vector with the administration of only a single prodrug, MeP-dR. Conclusion The bi-functional fusion suicide gene vector, pcDNA3.0/PNP-CD, yields powerful cytotoxic effects on HCC cells in the presence of the synergetic treatment of its specific prodrugs, which would be a high-performance therapeutic vector in gene therapy for liver cancer.

Streptozotocin-induced diabetic rats were treated with resveratrol for 12 weeks.Compared with group of diabetic rats without treatment,the levels of urine albumin,serum creatinine,and blood urea nitrogen were significantly decreased and pathological changes in kidney were improved in treatment group ( P＜0.05 ).The mRNA expressions of FoxO1 and catalase were higher and FoxO1 phosphorylation level in diabetic rats treated with resveratrol was lower than that in diabetic rats without treatment (P＜0.05),suggesting that resveratrol may protect the kidneys of diabetic rats via regulating expression of FoxO1.

ObjectiveTo determine whether antibiotic prophylaxis can reduce the risk of postoperative infective complications in men undergoing transrectal prostatic biopsy (TPB) who had sterile preoperative urine.MethodsMEDLINE, EMBASE, Cochrane Collaboration Reviews, Chinese Medical Current Contents (CMCC), and National Knowledge Infrastructure (CNKI) were searched for rando-mized controlled trials that compared the effect of antibiotic prophylaxis with placebo or active controls for men undergoing TPB with preoperative sterile urine. Two reviewers independently extracted the data of patient characteristics and outcomes based on a prospectively developed protocol.ResultsA total of 12 trials (3 placebo controlled, 3 non-treatment controlled, and 6 activly controlled) involving 1 987 patients, met the inclusion criteria. Prophylactic antibiotic use in patients at low risk undergoing TPB significantly decreased bacteriuria and middle degree fever incidence, but could not decrease the incidence of bacteremia. The relative risk for post-TPB bacteriuria, middle degree fever, and bacteremia were 0.32 (95% CI 0.23 to 0.46), 0.37 (95% CI 0.17 to 0.77), and 0.96 (95% CI 0.61 to 1.50), respectively. Effective antibiotic classes included quinolone, co-quinolone and nitroimidazole, and co-trimethoprim and sulfamethoxazole. Treatment protocols of any duration were effective.ConclusionAntibiotic prophylaxis obviously decreases the incidence of bacteriuria and middle degree fever but not bacteremia in men with preoperative sterile urine undergoing TPB. A significant decrease in bacteriuria incidence can be achieved with a range of antibiotic agents, including quinolones and co-quinolone and nitroimidazole. Treatment protocols of any duration are effective with no heterogeneity.

To investigate the mechanism of inhibitory effect of a novel bFGF antagonist peptide isolated from the phage display random heptapeptide library on cell proliferation induced by basic fibroblast growth factor. The effect of P7 on cell morphology was observed under an inverted microscope. Flow cytometry was applied to analyze the effect of P7 on cell cycle progress of bFGF-stimulated cells. The effect of P7 on bFGF-induced activation of MEK and Erk1/2 in MAPK pathway was detected by Western blotting. The results showed that no significant cell morphology change was observed in the range of detected concentrations of P7. Cell cycle analysis showed that P7 decreased S-phase cell population and arrested cell cycle at the G0/G1 phase of bFGF-stimulated cells. The results of MAP kinase activation assay indicated that P7 decreased bFGF-induced MEK and Erk1/2 phosphorylation in a dose-dependent manner. P7 inhibited proliferation of bFGF-stimulated Balb/c 3T3 cells possibly via cell cycle arrest at the G0/G1 phase and down-regulation of signal molecular activation in MAPK pathway.

Objective To discuss the method, therapeutic effect and safety of CT-guided 125I radioactive seed implantation for the treatment of cervical lymph node metastasis. Methods CT-guided 125I radioactive seed implantation was performed in 32 patients with pathologically proved cervical lymph node metastasis Results The local effective control rates of the cervical lymph node metastasis at one, 3, 6 and 12 months after the treatment were 81.3%(26/32), 84.4%(27/32), 93.7%(30/32) and 87.5%(28/32) respectively. Conclusion For the treatment of cervical lymph node metastasis, CT-guided 125I radioactive seed implantation is technically simple and clinically safe with reliable curative effect; this treatment is very effective in improving local tumor control rate.

Objective To establish the three-dimensional culture model of the human skin squamous-celled carcinoma ,and isolate the highly invasive cell subsets ,and compare their biological characteristics .Methods By using the composite chitosan tissue engi-neering skin ,the three-dimensional culture model of the human skin squamous-celled carcinoma was successfully established , through digestion ,we divided and got the high-invasive squamous carcinoma cells subsets .After that ,the different invasive proper-ties of subpopulations of cells in nude mice tumor effects were compared .Results Different concentrations of cinnamaldehyde and interferon on skin squamous-celled carcinoma cell proliferation inhibition had statistical significance (P<0 .01) ,the different invasive characteristics of skin squamous cells carcinoma between subpopulations of cells proliferation inhibition also was statistically signifi-cant(P<0 .01) ,and the two different invasive properties of subpopulations of cells in the tumor characteristics also had statistical significance(P<0 .01) .Conclusion We established the three-dimensional culture model of the human skin squamous-celled carcino-ma successfully ,by using this model ,we could screen the higher invasive cells subsets .

In this paper, the biological compatibilities of FGF/Collagen compound sponges are evaluated. Intracutaneous stimulation test and shortterm muscle embedding test of FGF/Collagen compound sponges made of different materials are performed. Experimental data are analyzed and evaluated according to the standard. The two sponges get marks of 0 in acute eye stimulation test and intracutaneous stimulation test, thus they are not stimulating to tissues. In shortterm muscle embedding test, they don't lead to inflammation reactions and can be degraded and absorbed in short term. FGF/Collagen compound sponge proves good biological compatibility and has a cheerful prospect in medicine.

Objective To systematic review bladder tumor antigen(BTA) stat in the diagnosis of bladder cancer. Methods MEDLINE, EMBASE, Cochrane Library, CMCC and CNKI were searched for studies about BTA stat in the diagnosis of bladder cancer. The search strategy was made according to the Collaborative Review Group search strategy. Data were extracted by two reviewers using the designed extraction form. The software MetaDiScl. 4 was used to review management and data analysis. Results Seventy-one relevant studies were searched, of which 20 studies were included with 5929 patients involved and 51 studies were excluded. Heterogeneity (except for threshold effect) was found within these studies. A meta-analysis was performed using random effect model. Pooled accuracy indicators like sensitivity, specificity, positive likelihood ratio (LR), negative LR and diagnostic odds ratio (dOR) were 0. 64(95%CI 0. 62-0. 66), 0. 72(95%CI 0. 70-0. 73) , 2. 33(95% CI 2. 03-2. 68), 0. 48(95%CI 0.42 -0.55) and 5.12(95%CI 3. 88-6. 75), respectively. The sensitivity increased with tumor grades. The sensitivity of primary tumors was higher than the recurrent tumors. Area under curve (AUC) of SROC curve was 0. 7500 and Q* index was 0. 6934. Conclusions The performance of urine BTA stat is moderate in the diagnosis of bladder tumor. It can be an optional non-invasive test and an important adjunct test in preoperative detecting and postoperative monitoring of bladder tumor.

In order to decrease the potential side-effects of human basic fibroblast growth factor (hbFGF) caused by its broadspectrum mitogenic activity, a single residue of hbFGF, the residue serine 108, was replaced with neutral alanine residue to construct a mutant of hbFGF (mhbFGF) with reduced mitogenic activity. The mutant was overexpressed in Escherichia coli BL21(DE3) by IPTG induction. The expression level of mhbFGF was about 30% of the total cellular protein. The expressed mhbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate. Measured by MTT method, the effect of mhbFGF on Balb/c 3T3 cell proliferation was much lower than that of wild-type hbFGF. The purified recombinant mhbFGF was prepared and sufficient for the following pharmacological and safety studies.

AIM:To study the mechanism of the unique export of one of human basic fibroblast growth factor (hbFGF) forms lacking the N-terminal nuclear localization signal (NLS),we high expressed and purified this hbFGF form in E.coli strain BL21(DE3).METHODS:The cDNA fragment of the hbFGF amplified by polymerase chain reaction (PCR) was cloned into the expression vector pET3c and expressed in BL21(DE3) by IPTG induction.The expressed hbFGF was purified by ionic exchange and heparin affinity chromatography from the supernatant of bacteria lysate.The mitogenic activity was measured by MTT.RESULTS:The expression level of hbFGF in E.coli was about 20% of the total cellular protein.The appreciable mitogenic activity of the purified hbFGF was comparable to that of hbFGF standard.CONCLUSION:The BL21(DE3)/ pET3c expression system could be used to high express hbFGF lacking NLS.The purified recombinant hbFGF was prepared and sufficient for further study.