Objective To establish a new atomic fluorescence spectrometry for the determination of As,Hg and Se in human gastric and intestinal juice. Methods The KBH4 solution was used as a reducing agent to generate hydrides of As,Se and metallic vapour of Hg from samples. Results The lineal range of the method was 5.00-80.00 ng/ml for As, 0.10-4.00 ng/ml for Hg, 0.50-5.00 ng/ml for Se respectively. The RSD was less than 6.0%. For As, Hg and Se, the limits of detection were 0.20, 0.01, 0.33 ng/ml respectively, the recovery rates were 98.6%-104.4%, 92.0%-104.0%, 98.5%-100.5% respectively. Conclusion The method is simple, rapid and accurate with satisfactory results.

Objective To construct of shRNA lentiviral expression vector targeting APRIL (aproliferation-inducing ligand) gene in CFPAC-1 cell of human pancreatic cancer. Methods We used gene engineering to screen RNA interference targeting sequence of APRIL gene. The complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed, synthesized and cloned into the pGCL-GFP vector. The resulting lentiviral vector containing shAPRIL were named LV-shAPRIL. Then it was conformed by PCR and DNA sequencing identification. 293T cells were eotransfected with LV-shAPRIL,pHelper 1.0 and pHelper 2.0 to product ientivirus. The titer of virus was tested according to the expression level of GFP in the 293T cells. After recombinant lentivirus infection into CFPAC-1 cells, we used real-time RT-PCR and Western blotting to examine APRIL mRNA and protein expression at different cell culture period.Results PCR analysis and DNA sequencing conformed that shAPRIL DNA was successfully inserted into the lentiviral vector. The titer of concentrated virus were 5 × 107 TU/ml. APRIL expression in CFPAC-1 cells were inhibited significantly at both mRNA and protein level. APRIL mRNA expression were decreased 73%, 70%and 71% , respectively, after the infection of 4 days, 4 weeks and 8 weeks by LV-shAPRIL. APRIL protein expression were decreased 66%, 63% and 62%, respectively , after the infection of 4 days , 4 weeks and 8weeks by LV-shAPRIL. Conclusions ShRNA lentiviral expression vector targeting APRIL gene has been successully constructed, and it can effectively inhibit the expression of APRIL gene in CFPAC-1 ceils. This study lays a foundatin for in vivo research APRIL gene scilence in pancreatic cancer cell using the model of nude mice.

Objective To explore the relationship between self-efficacy and self-management behaviors of patients with chronic obstructive pulmonary disease (COPD).Methods A total of 350 COPD patients form 5 residents' committees in shanghai were recruited by using a convenient sampling method and were scored using Chinese Self-efficacy Scale (CSES)and the self-management behaviors scale.Spearman's correlation analysis was performed to detect the relation of self-efficacy with self-management behaviors.Results Three hundred and twenty adults were included in this investigation,and their average FEV1/FVC was (57.86 ± 7.06)％,average score of self-efficacy was 74.2 ± 9.5.In self-management behaviors,time spent on physical exercise was (16.2 ± 33.9) minutes per week,and endurance exercises accounted for (109.0±49.0) minutes per week.The score of cognitive symptom management practice was 0.9 ± 1.0 and communication with physicians was 0.7 ± 0.8.Total score of self-efficacy was positively correlated with self-management behavior in each dimension (r values were 0.522,0.407,0.330 and 0.044,respectively ; all P ＜ 0.01).Conclusions Self-efficacy and self-management behaviors of COPD patients need to be improved,and self-efficacy may be related to self-management behaviors.

A method for the determination of methylmercury in seafood has been developed using dispersive liquid-liquid microextraction followed by direct mercury analyzer. Total mercury was detected by direct mercury analyzer, and inorganic mercury was calculated by the difference. The parameters affecting the extraction efficiency, including the selection of extractant and dispersant, their volume ratio, concentration of HCl and NaCl have been optimized in this study. The results showed that CH2 Cl2 as extractant, ethanol as dispersant, Volume ration of 1:5, 1 mol/L HCl and 120 g/L NaCl were chosen. The detection limit and the dynamic liner range were 0. 10 μg/L and 0. 2-20 μg/L, respectively. The relative standard deviation was 6. 0% for eleven replicates at the spiked level of 2. 0 μg/L. The enrichment factor was 8. For total Hg determination, the detection limit and the dynamic liner range for methylmercury were 0. 10 μg/kg and 0. 2-50 μg/kg, respectively. The relative standard deviation was 2. 4%. The method was simple, fast and a little solvent needed. Some certified reference materials were analyzed to validate the accuracy of the proposed method, and the results were in good agreement with the reference value. Besides, the method was applied to the real samples with satisfactory results.

Objective To measure and calculate the dose distribution (tissue absorbed dose) of mouth floor area while using 125I to treat sublingual gland carcinoma.Methods Phantom of head and neck was used to place the 125I radioactive seeds to simulate the sublingual gland carcinoma treatment.125I radioactive seeds of 29.6 and 25.9 MBq per seed were used as two groups,with 31 seeds in each group,and prescribed dose (peripheral matched dose) was 120 Gy.Thermoluminescence dosimetry (TLD) was used to measure the absorbed dose value in the simulated target and adjacent area.Gafchromic Eriochrome Black T (EBT) film was used to draw the dose distribution curve.Results Dose absorbed in the target area,target area center and the adjacent area one centimeter away from target reached 160 Gy,390-500 Gy,and 90-170 Gy,respectively.Dose of the skin ranged from 25 to 81 Gy,dose of mandible ranged from 7.9 to 67 Gy.No radiation cold spot was found.Conclusions 125I seeds could achieve an effective therapeutic dose distribution of the target area for sublingual gland carcinoma.Dose absorbed in the adjacent tissue is under safety limit.The radiation dose at mandible is lower,reducing the possibility of radiation damage to the bone.

Objective To construct small hairpin RNA(shRNA) lentiviral vectors targeting human a proliferation-inducing ligand(APRIL) gene and detect the titer of virus and suitable multiplicity of infection (MOI) after 293T cells were infected by the lentival vectors. Methods Three RNA interference targeting sequences of APRIL gene were screened including shAPRIL1210, shAPRIL1754 and shAPRIL1604. Both sense and antisense Oligo DNA of the targeting sequences were synthesized and cloned into the pGCL-GFP vector, respectively. The resulting lentiviral vectors containing shAPRIL were named LV-shAPRIL1210, LV-shAPRIL1754, LV-shAPRILI604. Then they were confirmed by PCR and DNA sequencing. 293T cells were co-transfected with LV-shAPRIL, pHelper 1.0 and pHelper 2. 0 to product lentivirus, respectively. The titer of virus and suitable MOI were tested according to the expression level of GFP in the 293T cells. Results PCR analysis and DNA sequencing confirmed that three shAPRIL DNA were successfully inserted into the lentiviral vectors. The titers of concentrated virus were 5 × 107, 6 × 107 and 4 × 107(transduction units )TU/ml, respectively, and the suitable MOI was 5. Conclusions Three shRNA lentiviral vectors targeting human APRIL gene have been successfully constructed, which lays a foundation for future studying APRIL's gene silencing in related target cells.

Objective To investigate the inhibitive effect of shRNA (short hairpin RNA) targeting APRIL gene on the pancreatic cancer cells in vitro and in vivo, in order to explore the feasibility of gene therapy for pancreatic cancer. Methods The LV-shAPRIL targeting APRIL gene had been constructed before, and was used to infect the CFPAC-1 cells. Cell proliferation and apoptosis were examined by MTT and flow cytometry. Then CFPAC-1 cells were used to construct the model of transplantation tumor into the nude mice, the tumor growth was assessed after LV-shAPRIL treatment. Results 96 hours after the LV-shAPRIL infection into CFPAC-1 cells, the cell proliferation was significantly inhibited when compared with control group and lentivirus infection group (P＜0.05 ). Flow cytometry showed the apoptosis ratio of the CFPAC-1 cells was (17.35±0.96)% in LV-shAPRIL group, which was higher than that in control group and lentivirus infection group (P＜0.05 ). After LV-shAPRIL injection into the model of nude mice, the tumor growth was slower than that in the two control groups. The tumor's volume of the LV-shAPRIL group was(821.8±123.3) mm3 and the mass was (2.16±0.18)g at 27 day, and were obviously depressed, when compared with two control groups (P±0.05). Conclusions LV-shAPRIL targeting APRIL gene can inhibit the growth of the CFPAC-1 cells in vitro and vivo. This may provide a new gene therapy approach for pancreatic cancer.

Objective To investigate the relationship between autophagy and metastasis of nasopharyngeal cancer ( NPC) cell lines 5?8F and 6?10B after X?ray irradiation and the related mechanism. Methods Two substrains, 5?8F and 6?10B, of the NPC cell line SUNE1, with high and low metastatic potentials, respectively, were used in our study. After 4 Gy X?ray irradiation, 5?8F cells were treated with rapamycin ( 20 μmol/L) to induce autophagy and 6?10B cells were treated with LY294002( 10μmol/L) to inhibit autophagy. The autophagy and metastatic activity of NPC cells were determined using qRT?PCR, Western blot, Transwell assay, laser confocal microscopy, and transmission electron microscopy. Results 5?8F cells showed a lower level of autophagy than 6?10B cells after X?ray irradiation. Rapamycin increased the autophagy and inhibited the metastasis of 5?8F cells after irradiation, while LY294002 inhibited the autophagy and increased the metastasis of 6?10B cells. Conclusions NPC 5?8F cells, which have a high metastatic potential, have a lower level of autophagy than 6?10B cells, which have a low metastatic potential. Autophagic inhibition could increase the metastatic activity of NPC cells, while autophagic activation could reduce their metastatic activity. Mechanistic analysis indicates that the PI3K/AKT/mTOR pathway is involved in this process.

Objective To investigate the self-efficacy levels and its influencing factors of community-based patients with chronic obstructive pulmonary diseases (COPD).Methods From October 2008 to March 2009,320 community COPD patients were recruited from a Shanghai community.They undertook questionnaires,scale survey and pulmonary function testing so as to investigate the influencing factors of self-efficacy.Results The total scale of self-efficacy was 74.24 ± 9.50 and the level of selfefficacy in 286 cases( 89.4％ )was intermediate.The knowledge of COPD,social supports,forced expiratory volume at 1 second (FEV1)/forced vital capacity (FVC) and self-management level were entered into regression equation and could explain 57.1％ of the total variance of independent variables.Conclusions The knowledge of COPD,social supports,FEV1/FVC and self-management level are the major influencing factors of self-efficacy in the COPD patients.We should improve the knowledge of disease and strengthen the psychological care and social supports so as to improve their quality of life.

OBJECTIVE To investigate the relation of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) expression with cervical lymph metastases in papillary thyroid carcinoma. METHODS The expression of COX-2 and VEGF in 79 specimens of papillary thyroid carcinoma were evaluated with SP immunohistochemical methods. In all the 79 cases, there were 46 cases with cervical lymph node metastases and 33 cases without cervical lymph node metastasis. RESULTS The positive expression rates of COX-2 and VEGF in the cases with cervical metastases were 81.6 % and 86.8 % respectively, and in the cases without cervical lymph metastases were 54.5 % and 66.7 % respectively. There was a significant difference in the positive expression rates of COX-2 and VEGF between two groups (P