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.

ObjectiveTo observe the effect of probucol combined with atorvastatin in treatment of stenosis carotid atherosclerosis.MethodsEighty-six patients with stenosis carotid atherosclerosis were separated into two groups with 43 cases each by random digits table.The patients were treated with probucol combined with atorvastatin in observation group and atorvastatin only in control group.The changes of arteriosclerotic plaque and blood fat pre- and poet-treatment were observed.Results There was no significant difference in the levels of lumen area (LN) and intima-media thickness (IMT) pre-treatment between two groups (P＞ 0.05).Compared with pre-treatment,the levels of LN and IMT post-treatment were significantly decreased in observation group(P ＜ 0.05 ),while no significant difference in control group (P ＞0.05).There was significant difference in the levels of LN and IMT post-treatment between two groups [(0.59 ±0.19) cm2 vs.(0.72 ±0.28) cm2,(2.76 ±0.41 ) mm vs.(3.27 ±0.57) mm](P＜ 0.05).There was no significant difference in the levels of total cholesterol (TC),triacylglycerol (TG),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C) pre-treatment between two groups (P ＞0.05).Compared with pre-treatment,the levels of TC,TG,LDL-C post-treatment were decreased,HDL-C was increased,there were significant differences (P ＜ 0.05 ).Post-treatment,the levels of TC,TG,LDL-C in observation group were lower and HDL-C was higher than those in control group (P ＜ 0.05).Conclusion Probucol combined with atorvastatin in treatment of stenosis carotid atherosclerosis can improve arteriosclerotic plaque and low blood fat,which can be applied in clinic.

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.

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.

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.

ObjectiveTo analyze the systolic function of subendocardial myocardium of the left ventricle in patients with diabetes mellitus (DM) and DM associated hypertension by speckle tracking imaging (STI).Methods The left ventricular ejection fraction(LVEF) and left ventricular mass index (LVMI) of all subjects including 30 patients with diabetes mellitus(DM group),25 patients with DM associated hypertension (DH group) and 30 healthy volunteers (control group) were normal and near.The common parameters of echocardiogram were measured and the peak systolic longitudinal strain of subendocardial myocardium were analyzed from the long-axis view at the levels of mitral annulus,papillary muscle and apex by STI.All above parameters were compared among three groups.ResultsNo significant difference in all parameters was found between the control group and two patient groups,but the ratio of E/A was significantly lower in the two patient groups than that of control group ( P ＜0.01).The peak systolic longitudinal strain of some segments were significantly lower in the two patient groups than that of control group ( P ＜0.01).The peak systolic longitudinal strain of the subendocardial myocardium of all segments in the two patient groups were different decrease compared with the control group( P ＜0.01 or P ＜ 0.001).The peak systolic longitudinal strain of the subendocardial myocardium of the DH group except some segments were decrease compared with the DM group ( P ＜0.01).ConclusionsSTI may be early used to identify the left ventricular systolic function in patients with diabetes mellitus.

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.

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.

Chemokine is a small protein which plays an impor-tant role in men's physiological function.It has chemotactic ac-tivity and is often secreted by immune cells and glial cells like microglia or astrocytes.Through the effect of chemokine recep-tors on target cells,various immune cells can achieve directional migration and play an important role in the diseases related with immunity and inflammation.CCL2,also known as monocyte chemotactic protein-1 (MCP-1 ),is one member of chemokine CC subfamily (βsubfamily).It can chemokine monocytes, macrophages and T lymphocytes to affect their phagocytosis func-tion and produce antibodies to combat invading microorganisms. In recent years,it has been found that CCL2 plays a key role in the occurrence and development of the problems concerning cen-tral nervous system and immune system as well as cancer, AIDS,leukemia,diabetes and other diseases.This thesis is to give an elaboration on the latest research on CCL2 and the rele-vant diseases.

Unilateral facet dislocation of the lower cervical spine is an important subgroup of cervical spine injuries,accounting for from 12％ to 16％ of all cervical spine injuries.Unilateral facet dislocation occurs the most commonly at the C5-6 level,and is closely related to the anatomic and morphometric features of the cervical facet.This injury has been attributed to combined flexion and rotation,but it remains unclear whether flexion or rotation is the component of greater importance.Its treatment includes nonoperative and operative measures but optimal treatment has not been established.It is generally accepted that a recommendable treatment protocol begins with closed reduction and proceeds depending on whether the closed reduction succeeds and traumatic disc herniation is present or not.