A method was developed for the determination of 39 polybrominated diphenyl ethers(PBDEs) from mono- to hepta-brominated in coastal sediment by gas chromatography/mass spectrometry. The sample was extracted with a solution of dichloromethane: n-hexane( 1:1,V/V) ,followed by ultrasonic extraction for 60 min at 25℃. The clean-up step was optimized using silica gel and alumina. The detection limit of the method was in the range of 0.003 - 0.10μg/kg. The results indicate that the average recovery of the internal standard was from 66.2% to 118. 6%. The relative standard deviation RSD( % ) was from 0. 8 % to 18. 2%. The method for the detection of 39 PBDE congeners in coastal sediment was sensitive,high reproducible with satisfactory recoveries.

Objective To investigate the induction of specific anti-tumor immune response by transfected dendritic cells (DCs) with survivin mRNA of human pancreatic cancer, and to provide the experimental evidences for the treatment of human pancreatic cancer with DCs vaccine. Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs). After being transcripted and amplified, survivin mRNA was transfected into DCs by electroporation. The expression of survivin in DCs at different time points was detected by quantitative real-time PCR. The survival rate of DCs before and after transfection was determined by MTT method. The induction of specific cytotoxic T lymphocyte (CTL) response by survivin mRNA transfected DCs was measured by 51Cr standard cytotoxicity test. The induction of specific CTL activation by survivin mRNA transfected DCs was evaluated through testing released IFN-γ by ELISA method. Results After survivin mRNA transfection for 48h, the expression of survivin mRNA in DCs reached the highest point (46.09±6.57). After transfection, the survival rate of DCs was stabilized around 80%. The DCs transfected with survivin mRNA could effectively induce HLA-A2+ / survivin+ specific CTL immune responses. Stimulated with pancreatic cancer cell line Capan-2 cells or SCL-1 cells as control group, the IFN-γ released in 24 hours by survivin specific CTL were (28.79±5.70) U/ml and (25.12±2.13) U/ml respectively, there was no significant difference (P=0.761). Conclusion The induction of CTLs by DCs transfected with human pancreatic cancer survivin mRNA could produce specific anti-tumor immunity.

AIM：To introduce evidence-based pharmacy in respect to the definition,contents and principle in applying to decisions of clinical pharmacotherapy.METHODS：We focused the discussion on a brief summary of different study designs,factors affecting the strength of scientific evidence,and interpretation of the data.RESULTS：These application principles are helpful to guide clinical pharmacists solving clinical problems.CONCLUSION：Clinical pharmacist should use concepts in the evidence-based practice when making pharmacotherapeutic decisions in the future.

Objective To assess the pulmonary function before liver transplantation in patients with end-stage liver disease. Methods One hundred and fifty-four patients with end-stage liver disease, who were waiting for liver transplantation in our hospital, were enrolled into the study. The pulmonary ventilation function,small airway function and diffusion capacity were measured and analyzed respectively. Results Among 154 subjects,140 (90. 9%, 140/154) patients had abnormal pulmonary function, shown as pulmonary diffusing capacity reduction;followed by restrictive ventilatory function reduction (42. 8% ,66/154) and small airway function reduction (37. 7%, 58/154 ), the least common manifestation was obstructive ventilatory function reduction (28.6 % ,44/154 ). Conclusion Abnormal pulmonary function in patients with end-stage liver disease is common, and the pulmonary function tests before liver transplantation has certain referential value for pulmonary function damage evaluation and postoperatively respiratory tract management.

Objective To investigate the best method of transfecting total RNA extracted from pancreatic cancer MiaPaCa-2 cells into dendrtic cells (DCs). Methods DCs were cultured from peripheral blood mononuclear cells induced by rhGM-CSF, rhIL-4 and TNF-α. Morphology of DCs was observed. Flow cytometry was used to detect the mature DCs specific surface markers:CD40, HLA-DR, CD83, CD86. Mixed lymphocyte (MLR) was used to determine the ability of DCs to stimulate allogeneic T cell proliferation.Liposomal transfection, electroporation method and passive transfection was used to transfect MiaPaCa-2 cell total RNA into DCs, Real time RT-PCR and MTT assay was used to determine the expression of MUC1 mRNA and the survival rate of the RNA transfected DCs. Results The cells acquired showed typical DCs morphology, the positive rate of CD40, HLA DR, CD83 and CD86 were 34.3% ,50.2% ,89.2% and 73.6%,and they showed a strong ability to stimulate allogeneic T cell proliferation. 48 h after transfection with MiaPaCa-2 cells total RNA by using electroporation, the MUC1 mRNA amount (45.39 ± 9.33) in DCs was higher than that of liposomes method (3 1. 68 ± 7.25) and passive transfection method (18.53 ± 3.26) . DCs survival rate was (80.36 ± 2.43)% by using electroporation, which was relatively lower than (91.48 ±5.42) % by using passive transfection method, but higher than (67.44 ± 2.51) % by using liposomes method,and it was stabilized around 80%. Conclusions Transfecting total RNA extracted from pancreatic cancer MiaPaCa-2 cells into DCs with electroporation is efficient and safe.

Objective To investigate the efficacy of metholrexate combined with rmifepristone and laparoscopic surgery for treating ectopic pregnancy.Methods 85 patient with ectopic pregnancy were selected.41 cases treated with methotrexate and mifeprisione were considered as the control group,while 44 cases with laparoscopic surgery were considered as the observalion group on the voluntary basis.The clinical efficacy,serum β-HCG value and recovery time after 7d were compared between the two groups.Results After the treatment,the cure rate in the observation group was 100％ which was significantly higher than 90％ of the control group ( P ＜ 0.05 ),the serum β-HCG value of the observation group was lower and the restoration normal time was shorter than the control group ( P ＜0.05).Conclusions Laparoscopic surgery and conservative treatment of methotrexate combined with mifepristone are effective in the treatment of ectopic pregnancy.However,the clinical efficacy of the laparoscopic surgery is better and it has rapid recovery time,and it can completely cure the patients.

This study was set up to observe the changes in stress and immune functions in upper abdominal surgery after general anesthesia.In thirty elective upper abdominal operation patients,general anesthesia was induced with propofol,fentanyl,scoline and maintained with isoflane and atrumium. Catecholamines(CA),cortisol,interleukin-2(IL-2) and soluble interleukin-2 receptor(sIL-2R) were measured before anesthesia,after induction of anesthesia,at the end of operation,and on the first and third postoperative days.The results showed that epinephrine(E),norepinephrine(NE) concentration increased at the end of operation and on the first postoperative day(P<0.01)，Cortisol concentration decreased after anesthesia induction(P<0.05)，IL-2 concentration decreased at the end of operation(P<0.05)，and sIL-2R lever did not change.The investigation suggested that general anesthesia with propofol,fentanyl and isoflane can suppress CA and cortisol increase resulting from induction of anesthesia. There was no change in IL-2 level.However it can not suppress the increase of stress hormones and decrease of IL-2 due to operation.The stress response correlate with immune functions.

Objective To study the diagnosis and treatment of iatrogenic bile duct injury(IBDI).Methods The clinical data of 92 patients with IBDI who had been admitted to our hospital from January 1994 to December 2007 were retrospectively analyzed.Results The diagnosis of IBDI was based on the clinical findings,diagnostic abdominocentesis and imaging examination.Sixteen patients gave up treatment because of economic reasons,and 2 patients who were complicated with sepsis and multiple organ dysfunction syndrome died shortly after being transferred to our hospital.Forty-eight patients were treated by Roux-en-Y choledochojejunostomy,14 by endoscopic treatment.2 by bile duet repair+T tube drainage,3 by percutaneous catheter drainage,1 by choledochoduodenostomy,4 by percutaneous transhepatic cholangiodrainage,and 2 by ultrasound guided percutaneous puncture drainage.Sixty-two patients were followed up for 4 month to 10 years(mean,3.6 years),and satisfactory results were obtained.Conclusions Detection of IBDI during operation and prompt repair yield favorable outcome.Roux-en-Y choledochojejunostomy is the best choice for bile duct injury complicated with biliary stricture.Endoscopic and interventional treatments are important auxiliary measures in the treatment of bile duct injury.

Objective To compare and evaluate the oncological and functional outcomes between reserving thyroid partial laryngectomy and traditional partial laryngectomy for the treatment of stage T2-3glottic laryngeal carcinoma.Methods One hundred and twenty-eight patients treated from January 1999 to January 2009 were selected from all glottic laryngeal carcinoma patients with stage T2-3,who underwent surgery combined with radiation therapy.A retrospective review was randomized and matched by tumour subsite,TNM stage and age,which were confirmed by pathology and observation without thyroid perichondrium invaded or only local invasion of thyroid cartilage,there was no need to extensively resect laryngeal cartilages.Patients were divided into reserving thyroid partial laryngectomy group(study group) and traditional partial laryngectomy group (control group).Each group included 64 cases,which included T2N0 38 cases,T2N1 6 cases,T2N2 1 case,T3N0 17 cases,T3N1 2 cases.Local control rate,decannulation rate,the 3-year and 5-year accumulative survival rate were compared between two groups.Results The 3-year and 5-year accumulative survival rate were 89.8％ and 84.3％ in study group,91.5％ and 85.4％ in control group,there was no sognificant difference (x2 =1.687,P ＞ 0.05).The local control rate was 91.7 ％ (55/60) in study group and 93.2 ％(55/59) in control group,there was no significant difference (x2 =0.103,P ＞0.05).The decannulation rate was 98.3％ (59/60) in study group and 89.8％(53/59) in control group,there was significant difference (x2 =4.933,P ＜0.05).Conclusions Comparing with traditional partial laryngectomy,reserving thyroid partial laryngectomy is successful for treating properly selected stage T2-3 glottic laryngeal carcinoma.This operation has higher decannulation rate and lower complication,which is effective for reducing surgical invasion and facilitated the resumption of respiratory.

Objective To compare the distribution patterns and analyze the antimicrobial resistance of pathogens of different clinical samples .Methods All pathogens were isolated from clinical specimens(blood ,urine and wound) ,the distribution of patho-gens and drug resistance were monitored continuously from October 2012 to October 2013 in Daling hospital .Results 279 strains weredetectedfrombloodculture.Thedistributionincidenceof Escherichiacoli,Klebsiellapneumoniae,coagulasenegativeStaphy-lococci(CNS) ,Staphylococcus aureus ,Enterococcus f aecalis were 28 .67% ,12 .19% ,13 .62% ,9 .68% and 8 .60% respectively .574 strains were detected from urine culture .The distribution incidence of Escherichia coli ,Candida ,Pseudomonas aeruginosa ,K leb-siella pneumoniae ,Enterococcus f aecalis were 38 .68% ,18 .64% ,7 .84% ,7 .49% and 3 .66% respectively .292 strains were detected from wound secretion .The distribution incidence of Staphylococcus aureus ,Escherichia coli ,CNS ,Pseudomonas aeruginosa ,En-terococcus f aecalis were 17 .12% ,14 .38% ,10 .27% ,7 .53% ,5 .48% respectively .Conclusion Clinical using of antimicrobial agents should monitor drug resistance and consider various factors .