Objective To construct dendritic cell(DC) vaccine expressing hepatitis C virus(HCV) multi-cytotoxic T lymphocyte(CTL) epitopes,which can stimulate T-cell response in vitro,providing certain information for immunization experiments in vivo for the next step.Methods DCs were infected with recombinant defective adenoviruses expressing two HCV CTL epitopes with tagged green fluorescent protein(GFP).The rate of infection was detected by the fluorescence microscope or flow cytometry;the expression of multiple CTL epitopes in DC was proved by RT-PCR and Western Blot.The cell surface markers of DC such as CD80,CD83,CD86 and HLA-DR were identified by flow cytometry.T cell proliferation effect promoted by DC was observed by cell counting kit-8(CCK-8).IL-12p70 in DC culture supernatant and IFN-γ in T cell supernatant were detected by Enzyme-Linked Immuno Sorbent Assay(ELISA).HCV specific CTL activity was measured by LDH release assay.Results Recombinant multi-CTL epitopes and GFP were successfully expressed in DC.Adenovirus can promote DC maturation,the percentages of CD80,CD83,CD86 and HLA-DR were(71.19±3.29)%,(81.21±5.07)%,(91.23±4.24)%,(97.95±5.31)% respectively.Infected DC promotes homologous T cell proliferation and the stimulation index was 6.806±0.247 when DC:T was 1:10.The secretion of IL-12 and IFN-γ was also increased to(193.83±6.25) pg/ml and(111.14±2.09) pg/ml respectively.The CTL stimulated by infected DC could specifically kill Huh-7.5 cells transfected with FL-J6/JFH transcripts.The cytotoxicity was 35.99% when the rate of effector to target was 100:1.Conclusion The multi-CTL epitopes recombinant adenovirus can effectively infect DC in vitro,promoting the T cell immune response,laying the foundation for developing anti-HCV DC vaccine.

Objective: Three dimensional X-ray imaging by Volume RAD, familiar with the operation method, a reasonable set of parametersin time, improve the technology efficiency at the same time, the control of possible ways to reduce the radiation dose. Methods: The 168 cases of patients with Volume RAD examination were divided into low, middle, high dose group, dose ratio was 5:10:15; respectively to examine patient’s chest, head, cervical spine, pelvis and knee joints, observe its effect. To track acquisition and scanning sampling, analyze of experiment and the effect. Results:The experimental results show, small lung nodules and trabecular bone dose, high dose group in the image quality is better than the dose than the low dose group, the display image quality, no significant difference between the 3 groups. Conclusion:After a time exposure capture can be parallel to the image surface of the detector series, was displayed in thecomplex structure and the overlapping parts of the details. But its operations are complex, need the exposure dose on dozens of times, how to control the exposure time and dose, which needs further research to select the best dose control scheme.

Objective:To study the influences of estrogen and progesteron on bone histomorphometry and renal expression of 1,25-dihydroxyvitamin D_3 receptor(VDR) mRNA in ovariectomized rats.Methods: Twenty-five adult female SD rats were equally randomized into 5 groups:sham-operation(Sham),ovariectomization(OVX),ovariectomization plus estrogen treatment(OVX+E),ovariectomization plus progesteron treatment(OVX+P) and ovariectomization plus both estrogen and progesteron(OVX+E+P) groups.All animals were killed to assess renal VDR mRNA expression by RT-PCR after 3.5 months feeding.Undecalcified proximal tibia was used to assess the bone histomorphometry with digitizing image analysis system.Results: OVX group,with weakened bone structure,had decreased cancellous bone and increased parameters of osteal formation and resorption compared with Sham,OVX+E and OVX+E+P groups(P

Objective:To study the effect of glucocorticoid receptor beta overexpression on the regulatory effect of glucocorticoid on proliferation and differentiation of human osteosarcoma cell and to investigate the biological significance of glucocorticoid receptor beta.Methods: Glucocorticoid receptor beta was stably transfected into human osteosarcoma cells and the stable transfectants were treated with different concentrations of dexamethasone(10~(-10)-10~(-6) mol/L).Then cell proliferation was determined by MTT method and viable cell counting;alkaline phosphatase was detected by standard hydroxybenzene colorimetry.Results and Conclusion: In stable transfectants overexpressing glucocorticoid receptor beta,glucocorticoid-induced cell proliferation repression and cell differentiation are inhibited,indicating that glucocorticoid receptor beta may be an endogenous inhibitor of glucocorticoid receptor alpha.

Objective:To investigate clinical effects of a good control of blood sugar during perioperative period on surgical tolerance and postoperative recovery in elderly patients with gastric cancer and diabetes.Methods:A total of 92 elderly patients with gastric cancer and diabetes undergoing surgery in our hospital from August 2018 to March 2020 were randomly divided into a control group(taking conventional perioperative care, n=46)and an observation group(taking conventional perioperative care and good glycemic control, n=46). The changes of perioperative blood glucose level, surgical conditions and postoperative recovery in the two groups were observed and analyzed.Results:Fasting blood glucose level of observation group and control group was[(7.4±1.1)mmol/L vs.(7.9±1.3)mmol/L, t=-1.991, P=0.025]at 1 d before operation, [(7.2±0.9)mmol/L vs.(8.7±1.2)mmol/L, t=-6.782, P=0.000]during operation, and[(6.9±1.5)mmol/L vs.(8.5±1.1)mmol/L, t=-5.834, P=0.000]after operation.Anyway, fasting blood glucose level was lower in the observation group than in the control group.In observation vs control group, time of incision healing[(7.5±1.6)d vs.(9.7±2.2)d, t=-5.485, P=0.000], time to get out of bed[(3.3±0.8)d vs.(4.7±1.3)d, t=-6.221, P=0.000], length of stay in hospital[(10.5±2.7)d vs.(15.1±3.5)d, t=-7.058, P=0.000]were shorter in observation group than in control group.The incidence of postoperative complications was lower in the observation group than in the control group(8.70% vs.26.09%, χ2=4.484, P=0.028). Conclusions:The application of perioperative glycemic control can effectively maintain the perioperative blood glucose at a reasonable and stable level, improve the surgical tolerance and promote the postoperative recovery in elderly patients with gastric cancer and diabetes.And it plays a very positive role in ensuring the safety of surgery

Objective To explore the regulatory effect of 17β-estrodial on vitamin D receptor (VDR) expression in pre-osteoblasts and the involvement of MAPK signal pathway in the process. Methods MC3T3-E1 subclone 14 cells were cultured in phenol-red free medium supplemented with 10% fetal bovine serum (FBS). Serum free medium was used when the cells were experimentally treated. After the cells were treated with 17β-estrodial for pre-determined time periods, expressions of vitamin D receptor (VDR) mRNA and protein were determined by SYBR green-based quantitative PCR and Western blotting analysis, respectively, and the activation of MAPK in MC3T3-E1 cells was examined by Western blotting analysis. Then the changes of VDR mRNA and protein in MC3T3-E1 cells were detected after the cells were treated with MAPKs inhibitors and 17β-estrodiaL Results VDR mRNA and protein were upregulated in MC3T3-E1 cells after treatment with 17β-estrodial for 72 h. ERK/ MAPK signal in MC3T3-E1 cells was activated within 15 min after treatment with 17β-estrodial and the activation remained for 60 min; but it did not activate JNK and p38 MAPK pathways. 17β-estrodial-induced VDR upregulation in MC3T3-E1 cells could be partly inhibited by ERK/MAPK inhibitor U0126. Conclusion 17β-estrodial can upregulate VDR expression in osteoblasts and can rapidly activate MAPK signal pathway, which is involved in the estrogen-induced upregulation of VDR.

Objective: To study the effect of glucocorticoid receptor beta overexpression on the regulatory effect of glucocorticoid on proliferation and differentiation of human osteosarcoma cell and to investigate the biological significance of glucocorticoid receptor beta. Methods: Glucocorticoid receptor beta was stably transfected into human osteosarcoma cells and the stable transfectants were treated with different concentrations of dexamethasone (10-10-10-6 mol/L). Then cell proliferation was determined by MTT method and viable cell counting; alkaline phosphatase was detected by standard hydroxybenzene colorimetry. Results and conclusion: In stable transfectants overexpres sing glucocorticoid receptor beta, glucocorticoid-induced cell proliferation repression and cell differentiation are inhibited, indicating that glucocorticoid receptor beta may be an endogenous inhibitor of glucocorticoid receptor alpha.

Application of the techniques of pedicle screw fixation in t horacolumbar spine has forcefully promoted the development of spinal surgery. Th e key to the techniques is that the pedicle screw insertion must be located into the sole exact routeway of the three-dimensional-space, that is, into the inn er vertebral body through the pedicle according to the exact arrow plane angle a nd horizontal plane angle. Recently great development has been made in the resea rch on the techniques of pedicle screw insertion in thoracolumbar spine, especia lly in the supervising means during operation. The means have been developed fro m the traditional X-ray perspective or photograph apposition to the application of computer-assisted technique. This paper reviews and discusses the current d evelopment and prospects in the research on the techniques of pedicle screw inse rtion in thoracolumbar spine.

Objective To compare the clinical results of the intramedullary nails (Gamma nail and the proximal femoral nail) in treatment of intertrochanteric fractures of femur. Methods A review study was conducted on 116 intertrochanteric fractures of femur treated with the Gamma nail and 89 treated with PFN in our hospital between 2000 and 2003. Results In the Gamma nail group, the mean time for operation was 68.3(48 to 106)minutes,the mean blood loss during operation was 261(180 to 400)mL,the mean time for walking after operation was 5.0 (2.3 to 8.9)weeks and the mean time for bone union was 8.6(7.1 to 12.6)weeks. In the PFN group, the mean time for operation was 48.0(36 to 85)minutes, the mean blood loss during operation was 192(120 to 360)mL,the mean time for walking after operation was 5.3(2.5 to 8.1)weeks and the mean time for bone union was 8.8(6.9 to 12.1)weeks. There were significant differences in the mean time for operation and the mean blood loss between the 2 groups(P