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.

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.

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 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.

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.

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 investigate the eruption of cystic lesion associated impacted permanent teeth(CAIPT)of the jaws after fe-nestrating decompression in adolescents and to analyze the related factors that affect the eruption of CAIPT.Methods:32 cases of CAIPT were examined restrospectively by panoramic radiographs.The subjects were divided into erupted group(n =22),and nonerupt-ed group(n =10).Patient age,angulation of the CAIPT,the cusp depth,eruption space and the level of dental root formation meas-ured on intial panoramic radiograph,were subjected to statistical analysis.Results:The eruption of the impacted mandibular molar was significantly related to the cusp depth,level of dental root formation and the patient's age(P <0.05).The eruption of the impacted man-dibular premolar was significantly related to the cusp depth,level of dental root formation,the angulation of the tooth and the patient's age (P <0.05),but not to dental space for the eruption(P >0.05).Conclusion:The patient's age,angulation of impacted permanent teeth,cusp depth,eruption space,level of dental root formation should be considered in the management of CAIPT.

Objective To investigate the awareness rate of TCM knowledge, the application, and demand situation of Chinese appropriate techniques in health clinics in towns and townships of Hunan Province;To put forward the countermeasures of TCM continuing education. Methods 50%of the training doctors were adopted randomly from 4 training courses in health clinics in towns and townships in Hunan Province. The student number of each course was 49, 51, 53, 53, respectively. Self-designed questionaires were used to investigate 206 training doctors. Results 168 (81.55%) doctors’ degrees were technical secondary school degree and college degree;186 (90.29%) doctors’ professional titles were physician assistant and physician;The awareness rate of TCM knowledge was 63.83%;Different study backgrounds of doctors had different effects on the rate of TCM knowledge (χ2=42.42, P=0.01). The using rates of TCM appropriate techniques were all lower than 30%. 96.60%of doctors in health clinics in towns and townships hope to learn one or more TCM appropriate techniques to help diagnose and treat disease. Conclusion The degree, professional title, mastery of TCM knowledge, and TCM appropriate techniques were not optimistic. The doctors in health clinics in towns and townships showed strong desire for TCM appropriate techniques. The health department should combine the actual situation of doctors in health clinics in towns and townships and develop targeted TCM continuing education.