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 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.

Objective To evaluate the left ventricular mechanical dyssynchrony in patients with chronic heart failure(CHF)by two dimensional speckle tracking imaging(2D-STI).Methods 37 CHF patients were enrolled in this study.According to left ventricular ejection fraction(LVEF),the patienst were classified into two subgroups:group A,LVEF≤ 35 ％;group B,35 ％＜LVEF＜50 ％.35 healthy volunteers constituted the control group(group C).Standard 2D images were acquired in the 2-,3-and 4-apical views as well as the parasternal short-axis views at the level of the mitral valve and papillary muscles.The time to peak-systolic strain and strain rate were measured for each segment in the level of the mitral valve and papillary muscles in Qlab software.Dyssychrony parameters were defined as the standard deviation and the maximal time delay of the time to peak-systolic strain and strain rate for 12 segments.Parameters were compared among the three groups respectively.The correlation between dyssynchrony parameters and LVEF were analysed in CHF patients.The cut-off value of each parameter were derived from group C based on statistical principles.Dyssychrony rate of each parameter were calculated in group A and B,and compared within each group respectively.Results Compared with group C,all the dyssychrony parameters in group A were significant higher(P＜0.05),and part of the parameters(Tsl-12SD,Tsl-12Dif,Tslr-12SD,Tslr-12Dif and Tsr-12SD)in group B were remarkable higher(P＜0.05).Dyssychrony parameters in CHF patients correlated closely with LVEF(P＜0.05).Significant differences among dyssychrony rates were noted in both group A and B(P＜0.05),and Ts1-12SD showed the highest value in both subgroups.Conclusions 2D-STI is useful in quantifying left ventricular systolic dyssychrony in CHF patients,and the longitudinal parameters are more efficient to detect mechanical dyssychrony than the radial parameters and circumferential parameters.

Objective To investigate the effects of ginsenosides Rg 3 on vasculogenic mimicry of pancreatic cancer xenograft through the establishment of pancreatic cancer cell line SW -1990 subcutaneous xenograft model .Methods After pancreatic cancer xenograft in nude mice model beening established , All the mice were randomly divided into 4 groups and treated intraperitoneally ( IP) with saline and various concentrations (5,10,20 mg/kg) of ginsenosides Rg3.To observe the effect of ginsenoside Rg3 on tumor growth.Immunohisto-chemical-PAS staining was used to detect the effects of ginsenosides Rg 3 on vasculogenic mimicry of pancreatic cancer xenograft .and mRNA and protein expression of MMP 2、MMP9 were respectively evaluated by FQ -PCR and Western blot .Results The ginsenosides Rg3 can inhibit the growth of the tumor xenografts in nude mice .The inhibitory effect is the most obvious the 20 mg/kg of ginsenosides Rg3 group.The expression of MMP-2, MMP-9 were down-regulated compare with the control group , and the difference was signifi-cant;the Immunohistochemical -PAS staining showed the number of vasculogenic mimicry (+) and CD31 (+) were less than that in the control group .Conlusion Our results demonstrate that pancreatic vascular mimicry formation can be suppressed by Ginsenoside Rg 3 though reducing the expression of MMP -2, MMP-9 in our vivo experiments ,

Objective To explore the short-term clinical outcome of percutaneous posterior lumbar interbody fusion using B-twin stand-alone expandable spinal spacer in the degenerative lumbar diseases.Methods From January 2006 to March 2009,41 consecutive patients with lumbar degenerative disc diseases were enrolled and 44 intervertebral spaces were observed in this study.There were 20 males and 21 females,with an average age of 57.51(ranging from 31 to 77 years old).Indications for treatment included degenerative lumbar disc herniation(LDH) with intervertebral instability or Ⅰ? spondylolisthesis(25 cases),LDH with intervertebral space collapse(15 cases) and lumbar discogenic pain(1 case).Fusion segments included 3 cases of L3,L4 and L5,3 cases of L3 and L4,27 cases of L4 and L5,and 8 cases of L5 and S1.After percutaneous discectomy and endplate,curettage were meticulously carried out.A B-twin expandable spinal spacer was introduced into the intervertebral space and then expanded and an allograft cancellous bone of 4 to 5 g was implanted around the B-twin in the intervertebral space.The whole operation was monitored by C-arm fluoroscopy.The clinical outcome was evaluated by JOA scale and X-rays and CT examination before the operation,and 1 week,3 and 12 months after the operation.Results After follow-up of 3 to 38 months(mean 16 months),all the patients were evaluated,and excellent was obtained in 13 cases,good in 21 cases,fair in 5 cases,and poor in 2 cases.The excellent and good rate was 82.93% according to the JOA scale.X-rays and CT showed that the subsidence of the B-twin was found in 3 cases,and 1 case was significantly subsided by more than 20%.Cage shift was seen in 1 case.Conclusion Percutaneou posterior lumbar interbody fusion with B-twin is efficient with minimal surgical trauma,blood loss,better rehabilitation and reliable short-term outcome.Strict indications and pinpoint skills for surgery are the precondition to obtain satisfactory outcome in the treatment of degenerative disc disease.

Objective To evaluate the efficacy of minimally invasive percutaneous plate osteosynthesis(MIPPO) for the treatment of tibial fracture. Methods From August 2005 to January 2007,a total of 26 patients with tibial fracture were treated with MIPPO in our hospital.Among the patients,the facture was located at the proximal tibia in 8,the distal tibia in 7,and the shaft in 11.Results The incision size was 4.0-8.5 cm with a mean of 6.6 cm;the intraoperative blood loss ranged from 80 to 400 ml(mean,180 ml);and the operation lasted 40-120 minutes(mean,87 minutes).The swelling subsided 1-6 days after the operation(mean,2.5 days).All the incisions healed primarily.The patients were followed up for 6 month on average(3 to 15 months).The fractures healed in 3-13 months(mean,4.2) after the operation.No complications,including delayed incision healing,incision infection,parenchymal necrosis,bone exposure,delayed union,and non-union,occurred in the series.According to the Johner-Wruhs criteria,the knee and ankle functions were excellent in 19 patients,good in 4,fair in 2,and poor in 1.Conclusions The MIPPO provides biological fixation and results in minimal parenchymal injury in patients with tibial fracture.By using the technique,the patients recover quickly with few complications and mild reaction to the trauma.The method is an optimal approach that is worth being promoted.

Objective To investigate the effects of benazepril on urinary excretion of endothelin 1(Et 1) and the level of plasmic Et 1 (PEt) in patients with diabetes mellitus (DM) and diabetic nephropathy(DN).Methods PEt and 24 hours urinary excretion of Et 1 (UEtV) in 54 patients with DM and 39 patients with DN before and after taking benazepril (7.5～12.5mg/d) for two months were measured using radioimmunoassay method.Results PEt and UEtV in patients with DM were significantly higher than in normal control group ( P

By analyzing the quality requirements for the majors in pharmacy speciality and the shortcomings existing in present practice teaching this article brought up the corresponding improving measures,which include courses and teaching modes innovation,teaching ability improvement and management reinforcement.

Objective To explore the potential applications of a chamber for in vivo tissue engineering,and to establish a novel model for in vivo tissue-engineered cartilage.Methods Auricular chondrocytes were isolated,cultured and identified from the ears cartilages of New Zealand white rab bits; rabbit auricular chondrocytes(RACs) were seeded into the scaffolds:(1) RACs were seeded into collagen gel scaffold; (2) RACs were seeded into PLGA/collagen gel scaffold in vitro,and the compos ites were placed into the chambers and implanted in the donor rabbit.As control groups,the composites were implanted directly subcutaneously in the donor rabbit without using chambers,and the contents were harvested at 8 weeks after implantation.Gross examination,histologic and immunohistochemical staining and RT-PCR test were performed to evaluate the harvested contents.Results Under the same conditions inside the chambers,the contents formed into new cartilage-like tissue by histo logical and immunohistochemical staining and RT-PCR.In contrast,in the control groups without chambers displayed vascular invasion and inflammatory reaction in the subcutaneous layer of skin,which eventually led to fibrous tissue or absorption.Conclusions Cartilage is successfully constructed in an immunocompetent animal model using a bioinert perforated chamber.This method is effective in creating a relatively favorable environment for cartilage regeneration,which may provide a valuable reference for the clinical application of tissue regeneration.

Background and purpose:The incidence of hepatitis C virus(HCV) infection is increasing worldwide.This paper was to study the clinical features of HCV-related hepatocellular carcinoma(HCV-HCC).Methods:We retrospectively analyzed the clinical data from the patients with HCC who underwent curative liver resection and randomly selected 64 patients with HCV infection and 100 patients with hepatitis B virus(HBV) infection as a control group.Post-operative complications and early prognosis were studied.Results:As compared with the patients with HBV-HCC,average age,incidence of liver cirrhosis and complications were signifi cantly higher in the patients with HCV-HCC(P