Objective To investigate effect to kill primary breast cancer cells by dendritic cell (DC)with self-freeze thawing antigens of primary breast cancer cells co-cultured with cytokine induced killer cell (CIK). Methods Self-neoplasm antigen by primary cells of breast cancer in log phase growth was prepared and DC and CIK cells was cultured from peripheral blood. The CIK of co-cultured DC loaded self-neoplasm antigen was compared to PBMC, CIK cells with self-neoplasm antigen and the single CIK cells. Results Killing efficiency of PBMC, CIK, Ag-CIK and Ag-DC-CIK were (34.35±3.28) %, (45.91±2.78) %, (50.88±3.22) %, (62.10±5.94) %. There were significant difference between Ag-DC-CIK group and CIK-Ag and CIK group to primary cells of breast cancer (P <0.01). Conclusion The CIK induced from cocultured DC loaded with self-neoplasm antigen show a special killing activity to self-primary tumor cells.

Objective To analyze the superiority of implanting operation with autoskull stored at ultrahypothermia over low temperature's and frozen skull bone of oneself.Methods 42 case were observed and followed up after operation.Results Incision is healing by firct intention.there were many advantages,such as good biological activity,beautiful appearance,no repellent response,no immunoreaction and hard to infection,etc. After storing 1,3,6 and 12 months,the histological structure of frozen skull observed by electronscope was similar to those of fresh skull.Pestroyed skull bone cells were not found.All cases had no complication during the period of 3 to 12 months(mean 6.5 months) follow-up after operation.After 12 months,X ray and CT scan showed that skulls were healed.Conclusions Implanting operation with autoskull stored at ultra-hypothermia is one of the most effective technique for repairing skull defects.

ObjectiveTo study the feasibility of ex vivo gene transfer to donor heart by delivering i ntracoronarily a reporter gene (Lac-Z gene) to donor heart at the time of trans plantation.MethodsThe model of heterotopic heart transplantation in murine was established and the method of perfusing intracoronarily was performed. Twelve male BALB/c mice were divided into 2 groups randomly: gene-transfer group (group 1) and control grou p (group 2). In group 1, plasmid vector (PSV-?-gal containing Lac-Z gene)/li posome (DOSPER) was perfused intracoronarily into each donor heart at the time o f transplantation. In group 2, normal saline was perfused. Donor hearts were har vested 3 days after transplantation. Freezing sections were made for detection o f the transfer and expression of Lac-Z gene by histochemical staining (X-gal). ResultsThe expression of Lac-Z gene was detected in the donor hearts of group 1. In tw o donor hearts, the expression of Lac-Z gene was detectable in the myocardial c ells in the mid-layer of ventriculus; In one donor heart the expression was fou nd in the myocardial cells under epicardium. But no expression of Lac-Z gene wa s detected in donor hearts of group 2.ConclusionEx vivo gene transfer intracoronarily to donor heart in the form of plasmid vector /liposome complex at the time of transplant ation is feasible.

Aim To examine the role and uderlying mechanisms of Lin28 /let-7d axis in the proliferation of lung fibrobalsts and fibroblasts-into-myfibroblasts tran-sition,and provide novel strategy for the treatment of idiopathic pulmonary fibrosis (IPF).Methods We induced experimental lung fibrosis in mice by intratra-cheally injection of bleomycin (BLM).Ang Ⅱ and TGF-β1 were used to induce fibrogenesis in cultured MRC-5 cells;qRT-PCR and Western blot were applied to determine the changes of Lin28B,collagen 1 α1 and collagen 3α1 ;MTT assay,Edu satining and immun-ofluoresence were used to examine the cell viability, proliferation and fibroblasts-into-myofibroblasts transi-tion in MRC-5 cells.Results Lin28B was increased in the lung of mice with experimental lung fibrosis and in MRC-5 cells treated with AngⅡ or TGF-β1 .Moreo-ver,Lin28B enhanced collagen deposition via inhibi-ting expression of let-7d,which maybe contribute to the progression of IPF.In addition,further studies showed that Lin28B promoted proliferation and fibro-blasts-into-myofibroblasts in MRC-5 cells.Conclusion Lin28B /let-7d axis contributes to fibrogenesis via promotes fibroblasts-into-myofibroblasts transition, which may provide novel approaches for lung fibrosis treatment.

Objective To investigate the accumulation and maturation status of pulmonary conventional dendritic cells (cDCs) in the early phase of acute lung injury (ALI),and to explore the way of the inflammatory responses and lung injury modulated by cDCs in vivo.MethodsMale C57BL/6 mice were randomly ( random number) divided into the normal control group,6 h-ALI group and 24 h-ALI group.Murine model of ALI was made by intra-tracheal administration of lipopolysaccharide (LPS) and lung specimens were taken 6 h or 24 h later.The accumulation and maturation status of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate the lung inflammation.Transcription factors T-bet/GATA-3 mRNA ratio was determined to estimate the balance between Th1/Th2 responses.IFN-γand IL-4 were quantified to evaluate Thl-specific and Th2-specific cytokine production respectively.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Comparison between groups was performed using one -way ANOVA.ResultsCompared with normal control group,LPS challenge resulted in higher level of IL-6 and TNF-α,increased LWW/BW ratio and significant histopathological changes (P ＜0.01 ).The accumulation and maturation of pulmonary cDCs in 6 h-ALI group were significantly increased after LPS challenge (P ＜0.01 ),while the accumulation and maturation of pulmonary cDCs in 24 h-ALI group were significantly lower than that in 6 h-ALI group ( P ＜0.01 ).Compared with normal control group,the expression of T-bet mRNA in 24 h-ALI group was markedly enhanced ( P ＜ 0.01 ) and the production of IFN-γ was increased as well ( P ＜ 0.01 ).ConclusionsThe accumulation and maturation of pulmonary cDCs peaked within 24 h after LPS challenge,pulmonary cDCs may initiate and amplify acute lung inflammation of ALI by enhancing the Th1 immune response and ensuing cytokine production.

Objective To assess the diagnostic capability of spiral CT for giant pure seminoma in intraabdominal undescended testis.Methods Spiral CT of 6 cases with pure seminoma of inabdominal testes as proved by surgery and pathology were analyzed.All patients were male,and the age ranged from 31 to 45 years old with the mean of 35.2 years old.Results All tumors were located along the path of testicular descent on CT images.The arterial-supply of tumors all came from the testicular artery ipsilaterally.The draining vein could be seen between the mass and inferior vena cava or left renal vein in 5 cases.Isolaterally spermatic cord was absent in the inguinal region.Isolateral kidney was shifted upward.CT scans typically demonstrated a unilateral,mixed solid and cystic mass,with areas of solid located at the lateral aspect and areas of necrosis at the medial aspect.Contrast-enhanced CT scan showed mild enhancement of solid areas and band-like septal enhancement in areas of necrosis.There is no evidence of calcification or fat within the mass.Conclusion Spiral CT proves to be a very useful preoperative imaging modality for the giant intraabdominal seminoma.

Objective: To observe the effects of electroacupuncture on withdrawal anxiety in heroin-dependent patients. Methods: Sixty heroin-withdrawal subjects with negative urine morphine were digitally randomized into two groups: acupuncture group in which 30 cases were treated by electroacupuncture and control group in which 30 cases were given no treatment. The patients were assessed by Hamilton anxiety scale before treatment and one week, two weeks and three weeks after treatment. Results: After treatment, the anxiety symptoms of heroin-dependent patients were improved more significantly in the acupuncture group than in the control group (P<0.01). Conclusion: Electroacupuncture has good effects on anxiety symptoms and emotions of heroin-dependent patients.

Objective To clarify the role of FLT3 signaling-dependent pulmonary conventional dendritic cells (cDCs) in the pathogenesis of lipopolysaccharide (LPS)-induced acute lung injury (ALI),and as well as the modulation effects of cDCs in vivo on the inflammatory responses to acute lung injury.Methods Thirty C57BL/6 male mice were divided into normal control group,LPS group,FLT3L pretreatment group,lestaurtinib,(a high efficient and specific blocker in FLT3 signal pathway) pretreatment group and vehicle (DMSD) control group.FLT3L and lestaurtinib were administrated subcutaneously for 5 days.Murine model of ALI was subsequently established by intra-tracheal application of LPS and lung specimens were harvested 6 h or 24 h later.The accumulation and maturation of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate lung inflammation.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Lung myeloperoxidase (MPO) activity was measured to evaluate neutrophil infiltration.Transcription factors Tbet/GATA-3 mRNA ratio was determined to estimate balance of Th1/Th2 response.IFN-γ and IL-4 were quantified to evaluate Th1-specific and Th2-specific cytokine production respectively.Results The accumulation and maturation of pulmonary cDCs peaked at 6h after LPS challenge.FLT3L pretreatment significantly stimulated the accumulation and maturation of pulmonary cDCs (P ＜ 0.05),leading to markedly deterioration of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/GATA-3 mRNA ratio were elevated (P ＜ 0.05).Furthermore,the production of IL-6,TNF-α and IFN-γwas markedly increased by FLT3L pretreatment (P ＜ 0.05).In contrast,lestaurtinib pretreatment markedly inhibited the accumulation and maturation of pulmonary cDCs (P ＜ 0.05),leading to significant improvement of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/ GATA-3 mRNA ratio were decreased (P ＜ 0.05).Furthermore lestaurtinib efficiently suppressed the production of IL-6,TNF-α and IFN-γ (P ＜ 0.05).Conclusion This study thus demonstrated that FLT3 signaling-dependent pulmonary cDCs could control the initiation of acute lung inflammation response to LPS-induced ALI through the regulation of neutrophil infiltration and balance of Thl/Th2 response.

Objective To observe the efficacy of proparacaine hydrochloride in preoperative venipuncture. Methods Two hundred and furty patients hospitalized for preoperative venipuncture, between June 2015 to December 2015 in Jiangmen Central Hospital, were equally randomized into the intervention group and control group: the former was treated with proparacaine hydrochloride and the control group used traditional method. Visual analogue scale (VAS) was adopted to assess the effects of the anesthesia effect. At the same time the one-time success rate of puncturing and the adverse reactions were observed and compared between the two groups. Results Patients of the intervention group felt significantly less painful than that the control one (P<0.05). The successful rate of the intervention group was significantly higher than that of the control group (P<0.05). Conclusion Proparacaine hydrochloride is safe and effective for preoperative which reduces pain.

Objective To explore the effect of hepatitis B virus (HBV) X protein (HBx) on autotaxin (ATX) expression and its significance. Methods The recombinant eukaryotic expression vector of HBx ,pcD-NA3.1(+)-HBx,and the recombinant luciferase reporter gene vector of ATX promoter,pGL3-ATX,were con-structed and used to co-transfect HepG2 cells to examine the effect of HBx on the activity of ATX promoter. The sta-ble cell expressing HBx,HepG2.HBx,was constructed,and Western blot(WB)was used to detect the effect of HBx on ATX expression. Results The luciferase activity of pcDNA3.1(+)-HBx and pGL3-ATX group was 1.47 times as that of the empty vector cDNA3.1(+)and pGL3-ATX group(P<0.000). WB detection showed that the expression of ATX protein was increased in HepG2.HBx cells,and 1.75 times as that of HepG2 cells(P<0.05). Conclusion HBx can activate ATX promoter and up-regulate ATX expression ,thus suggests that HBV infection might enhance ATX/LPA signaling.