Objective To explore the effects of TMPRSS2 promoted suicide gene CD-5-FC combined with phytochemicals on the proliferation and apoptosis of prostate cancer (PCa) specific cell-line 22RV1.Methods From March 2016 to October 2016 TMPRSS2-VISA-CD/UPRT'vector was used to deliver pro-drug 5-FC (5-Fluorocytosine) in to PCa specific cell-line 22RV1.Transfection effect was verified by Western-blotting.5-FC,5-FU (5-Fluorouracil) and four phytochemicals epigallocatechin gallate (ECGC),Genistein,Daidzein,and Equol were selected in this study.MTS assay was performed to select dosages,which can kill 10％-40％ of 22RV1 cells,from their pre-set drug concentrations,respectively.According to their select concentrations,either 5-FC plasmid or 5-FU in combination with one of four phytochemicals were added in 22RV1 cells in orders,and then cultured them together.Besides,these four phytochemicals were added in 22RV1 cells individually regarding their selecting concentrations and cultured at the same time.Cell viability was detected by MTS assay on 24hrs,48hrs and 72hrs.Comparing the cell killing rates between the combination groups and each single drug groups on cell-line 22RV1.Results TMPRSS2-VISA-CD/UPRT'plasmid with 5-FC was transfected on PCa specific cell-line 22RV1 successfully.The most obvious transfection occurred on 48hrs.The selected concentration of the pro-drug 5-FC were 83.32 μmol/L and 833.20 μmol/L,5-FU were 76.87 mol/L and 768.70 μmol/L.The concentrations of four phytochemical agents were 2.00 μmol/L,10.00 μmol/L and 20.00 μmol/L.In term of the cell killing numbers,results showed that the combination treatment of 83.32 μmol/L 5-FC/ 76.87μmol/L 5-FU and 10.00 μmol/L/20.00 μmol/L four phytochemicals compared with 10.00 μmol/L/20.00 μmol/L single treatment of four phytochemicals,respectively,there was no significant difference (P ＞0.05),while the combination treatment of 833.20 μmol/L 5-FC/768.70 μ mol/L 5-FU and 2.00 μmol/L/10.00 μmol/L four phytochemicals compared with 2.00 μ mol/L/10.00 μmol/L single treatment of four phytochemicals,respectively,there was a significant difference (P ＜ 0.05).In addition,this study proved that 83.32 μmol/L 5-FC and 76.87 μmol/L 5-FU,833.20 μmol/L 5-FC and 768.70 μmol/L 5-FU reached the same cell killing effect,and there was no statistical difference (P ＞ 0.05).Thus the suicide gene transfection was successful,and its function was the same as the effect of cytotoxic drugs.Conclusions This study has proved that this plasmid with 5-FC suicide gene system can be successfully and efficiently transfected into PCa cells 22RV1.As the pro-drug of 5-FU,5-FC got similar treatment effect with 5-FU,and inhibited cell proliferation.There was no synergistic reaction of enhancing cell apoptosis after combined phytochemical drugs EGCG,Genistein,Daidzen,Equol with suicide gene therapy on 22RV1 cell-line.

Objective To investigate the effect and possible mechanism of curcumin on the proliferation and apoptosis of cell strain of bladder carcinoma.Methods In curcumin group,T24 cells were treated with curcumin of 1,10,100 ?mol/L respectively,while no curcumin treatment in control group.The growth,apoptosis,AKT1 protein activity and mRNA level,caspase 3 content of T24 cells were observed by MTT,TUNEL,RT-PCR,immunoblot and image analysis technology 12,24 and 48 h after the commencement of curcumin treatment.Results In the curcumin group,the inhibition rate,apoptosis and the content of caspase 3 of T24 cells increased significantly.The activity of AKT1 protein decreased,but the mRNA level of AKT1 showed no changes.Conclusion Curcumin can inhibit the growth of T24 cells and induce the cell apoptosis,which mechanism is related to the inhibition of AKT1 signal pathway of T24 cells.

Objective and fair clinical trials are the main methods for assessing the clinical significances of the experimental findings. The development of translational medicine highly relies on high-quality clinical trials as well as trial reports. Although the definition of"quality of clinical trials"and"quality of trial reports"differs from each other, they are closely related and can be consistent in most circumstance in the context of"scientific integrity". The quality of trial reports can be basically assessed by their internal and external properties. The quality of a randomized trial can be assessed by Jadad scale and Cochrane collaboration's tool for assessing risk of bias, and the quality of a non-randomized trial by risk of bias tool and Newcastle-Ottawa scale. However, since Jadad scale lacks appropriate appraisal of allocation concealment and is too simple in evaluating blind method, assessment of allocation concealment should be added. A more widely accepted approach for assessing the quality of random trials is the combination of Jadad scale and Schulz's approach to allocation concealment till recent years.For non-randomized cohort studies and case-control studies, Newcastle-Ottawa scale might be suitable at present time.

Objective To establish a method for the quality control of Semen Cassiae.Methods A high performance liquid chromatographic method was developed to establish the fingerprint of Semen Cassiae,and 32 samples from various batches of Semen Cassiae were analyzed.Cluster analysis and principal component analysis were applied to study HPLC fingerprint and chemical pattern recognition.Results The fingerprint of Semen Cassiae was set up.Conclusion The method could be used for the quality control and comprehensive evaluation of Semen Cassiae.

Objective To establish a method for quality control of Radix Paeoniae Alba.Methods A high performance liquid chromatographic method was developed to establish the fingerprint of Radix Paeoniae Alba and 47 samples from various batches were analyzed.Cluster analysis and principal component analysis(PCA) were applied to study on HPLC fingerprint and chemical pattern recognition method. Results The result of PCA and cluster analysis showed that the samples were divided into two types.The HPLC fingerprinting of Radix Paeoniae Alba,showing 10 characteristic peaks,was established from 47 batches of Radix Paeoniae Alba.Conclusion The method provides an academic reference for the quality control of Radix Paeoniae Alba.

Objective To establish a method for quality control of Rhizoma Chuanxiong.Methods A high performance liquid chromatographic method was developed to establish the fingerprint of Rhizoma Chuanxiong,and 23 samples from various batches were analyzed.Cluster analysis and principal component analysis were applied to studying HPLC fingerprint and chemical pattern recognition.Results The fingerprint of Rhizoma Chuanxiong was set up.Conclusion The method provides an academic reference for controlling the quality of Rhizoma Chuanxiong.

This study is to investigate the protein and mRNA expressions of pro-inflammatory and anti-inflammatory cytokines in U937 foam cells and effects of Ginkgo biloba extract (GbE) on the cytokines.U937 cells were cultured with different concentrations of GbE (0.1,1,and 10 μg·L-1),and stimulated by 100 mg·L-1 oxidized low density lipoprotein (ox-LDL) for 24 h.The expressions of interleukin-1β (IL-1β),tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in culture solution were detected by enzyme-linked immunosorbant assay (ELISA) and reverse transcriptase polymerase chain reaction (RT-PCR).The results showed that incubated with 100 mg·L-1 ox-LDL for 24 h,the U937 cells became foam cells,the protein or mRNA expressions of IL-1β,TNF-α,IL-10,and its receptor IL-10R in U937 foam cells were higher markedly than those in normal U937 cells.When the cells were pretreated with GbE (0.1,1,and 10 μg·L-1),the increases of IL-1β and TNF-α in U937 foam cells were remarkably inhibited,but IL-10 expression increased greatly.Especially when cells were pretreated with 10 μg·L-1 GbE,the protein and mRNA expressions of IL-1β and TNF-α were markedly lower than those in U937 foam cells.The protein expression of IL-10 and mRNA expressions of IL-10 and its receptor IL-10R were markedly higher than those in U937 foam cells.GbE inhibited production of pro-inflammatory cytokines IL-1β and TNF-α,but up-regulated the production of anti-inflammatory cytokine IL-10 and its receptor IL-10R in U937 foam cells,which might be related with its anti-atherosclerotic actions.

Objective To investigate the effect of TGF-β1 on epithelial mesenchymal transition in Hela cells of human cervi-cal cancer.Methods Hela cells of Human cervical cancer cultured in vitro were divided into the experimental group and control group.In the control group,Hela cells were cultured in serum-free medium without TGF-β1.In the experimental group,Hela cells were treated with different concentrations of TGF-β1 (0.01,0.10,1.00,10.00 ng/mL).The morphological changes of Hela cells of human cervical cancer were stimulated by TGF-β1 at different time points observed under an inverted microscope,while the expres-sions of mRNA and protein of E-cadherin and Vimentin in Hela cells were detected by semi-quantitative RT-PCR assay and cellular immunohistochemistry respectively.Results Comparing with the control group,Hela cells stimulated by TGF-β1 for 48 h began to have morphological changes.Mesenchymal morphology changes were observed obviously after 72 h.RT-PCR analysis showed that the expression of epithelial marker E-cadherin mRNA was down regulated,while the expression of mesenchymal marker Vimentin mRNA was increased and showed a concentration dependence after the stimulation of TGF-β1,Comparing with the control group, the difference was statistically significant (P < 0.05 ).After stimulation of TGF-β1 in Hela cells,cellular immunohistochemistry showed that the concentration of TGF-β1 increased,the expression of E-cadherin protein gradually decreased,and the expression of Vimentin protein gradually increased at the same time.Comparing with the control group,the difference was statistically significant (P <0.05).Conclusion TGF-β1 may induce epithelial mesenchymal transformation in Hela cells of human cervical cancer.

Objective To analyze the epidemiological characteristics for new detecting cases of leprosy in 2012, in Yunnan Province, and provide clue and foundation for prevention and treatment in leprosy control. Methods The date of diagnosed leprosy patients in 2012 in Yunnan Province were collected and analyzed by disease reporting information system. Results 230 new cases were founded in 2012, and the discovery rate was 0.50/100 000. 4.35% of new cases were children, 64.35% of new cases were MB and 19.57% had grade 2 disability. 17 recurrent cases were founded in 2012,and 6 of them had received MDT. By the end of 2012, there were still 1084 present case in Yunnan Province,the prevalence rate was 0.24/10 000,and 485 of them need MDT. Conclusion The prevalence of leprosy was decreased,and the prevalence varies a lot in different regions. Honghe and Wenshan are still the focus regions. Leprosy is still a serious public health and social problem in Yunnan Province. In order to reduce the burden of leprosy and eliminate the leprosy danger, long-term financing investment and prevention are still needed.

Objective To establish HPLC fingerprint for Fructus Aurantii Immaturus.Methods The HPLC method was used with Diamonsil-C18 column (250 mm?4.6 mm,5 ?m),and a mixture liquid of acetonitrile-0.01% NaH2PO4 as mobile phase in a gradient elution.The HPLC fingerprint for 36 batches of Fructus Aurantii Immaturus was studied on their similarity,cluster,and principal components analyses.The common HPLC fingerprint of Fructus Aurantii Immaturus was established,which was studied with principal components analyses.Results Under the selected spectrum condition,the 36 batches of Fructus Aurantii Immaturus were classified into two groups based on the result of similarity,cluster,and principal components analyses.Conclusion This method is reasonable and reliable to the quality control of Fructus Aurantii Immaturus.