Objective To investigate the depressing effect of antigene peptide nucleic acid (AGPNA)on the k-ras gene expression of human pancreatic cancer Patu8988 cells, the inducing apoptosis effect on Patu8988 cells, and the biodistribution characteristics in nude mice bearing xenografts using 188Re-k-ras-AGPNA.Methods The expression level of k-ras mRNA and the expression ratio of k-ras protein in Patu8988 cells transfected with AGPNA was measured by RT-PCR and flow cytometry ,respectively. The degree of cellular apoptosis 3 to 5 d after treating Patu8988 cells with 188Re-k-ras-AGPNA or 188ReO4- was determined by flow cytometry. For biodistribution study, 58 nude mice bearing Patu8988 cell xenografts were divided into two groups: intratumoral injection of 188Re-k-ras-AGPNA (Group A) and 188ReO4- (Group B). At different time points, the mice were sacrificed and organs of interest were excised, weighted and counted by a gamma counter. The organ uptake was calculated as a % ID/g and the absorbed doses of organs were calculated. One-way analysis of variance was used. Results After transfected with 1 nmol/ml AGPNA, the k-ras mRNA gray scale ratio and the expression ratio of k-ras protein were 1.00 ± 0.39 and (15.05 ± 5.07)%, respectively. They were significantly lower than those of the control group with 1.86 ± 0.07 and (24. 38 ± 5.40) % (F = 2. 545, 5. 327, P<0. 05). At 4 and 5 d after treatment in Group A, float cells' apoptosis ratios were (26.30 ± 7.45) % and (27.90 ± 10. 38) %, respectively. Tumors were the major distribution site in Group A with uptake of (37.47 ±21.31), (35.96 ±7.80) and (15.46 ±4.93) %lD/g at 1 h, 1 d and 7 d after intra-tumor injection, respectively. The absorbed dose of tumor was 15 569 mGy/MBq. Condusions Transfection with k-ras-AGPNA on Patu8988 cells may inhibit k-ras expression at mRNA and protein expression level, and 188Re-k-ras-AGPNA can induce apoptosis of Patu8988 cells.Tumor is the major distribution site in nude mice bearing human pancreatic cancer xenografts after intratumoral injection of 188Re-k-ras-AGPNA.

Catheter Ablation ; methods ; Humans ; Hypertension ; surgery ; Renal Artery ; Sympathectomy ; methods

Objective To perform a biomechanical study on and compare the stability of internal fixation with closed Kirschner wire in different spots from the epicondylus medialis to sulcus nervi ulnaris and epicondylus lateralis of humerus in the treatment of GartlandⅢsupracondylar fractures of humerus in children in order to provide basis for avoiding cubitus varus in clinic.Methods Twenty models of su- pracondylar fractures were made from ten humerus samples,then divided into two groups(group A and group B).They were fixed with closed Kirschner'wires.Ulnar Kirschner wire varied.In group A five spots were selected averagely from the epicondylus medialis to the sulcus nervi ulnaris.Kirschner wires entered from one of the five spots into another spot of humerus.In group B Kirschner wires entered defferent spots of humerus from the central spot of sulcus nervi ulnaris.Biomechanical tests were carried out,such as axial load,axial displacement,stress intensity,axial rigidity and anti-twisty ability.Forty-six children with Gart- landⅢsupracondylar fracture of humerus were treated and followed up for more than one year.Results In beth groups,with the angle of Kirschner wires and fracture side increasing,the strain and the vertical displacement decreased,the stress intensity and the axial rigidity increased,and the anti-twists ability was enhanced.There was statistic difference(P＜0.05).The follow-up of the 46 patients showed the elbow joint function recovered well with no cubitus varus malformation.Conclusion Internal fixation with closed Kirschner wires in sulcus nervi ulnaris from the epicondylus medialis and the epicondylns lateralis of humerus for GartlandⅢsupracondylar fracture of humerus in children has the advantages of enhancing bio- mechanical stability,effective prevention of secondary replacement of outlying black of the fracture and a- voidance of cubitus varus.It can be widely used clinically.

Objective To evaluate the relationship between combined muhigene detection and response to chemotherapy and prognosis in epithelial ovarian carcinomas(EOCS).Methods A total of 80 ovarian tissue samples taken from the surgical specimens of patients with EOCS of our hospital in the last two decades who had received chemotherapy "after surgery were paraffin-embedded.The samples were divided into 2 groups,good prognosis group(patients who survived more than 2 years,n=46)and poor prognosis group(patients who survived less than 2 years,n=34).The expression levels of ToPo-Ⅱ,Ki-67,MGMT, PCNA,p27,p53,pl6,P-gp,LRP,GST-?,bcl-2,C-myc,Fas,bax,MSH2,MRP and BCRP were investigated by the combination of tissue arrays and immunohistoehemical streptavidin-biotin peroxidase(SP) method in all samples.Data were analysed with SPSS 12.0 for windows.Results There were statistically significant differences in the positive expression levels of P-gp,BCRP,MGMT,MSH2,p27 and p16(62%, 50% and 50% in poor prognosis group vs 33%,28% and 28% respectively,P

Objective To isolate and identify the adult neural stem cells from the parenchyma of spinal cord in adult mouse.Methods The parenchymal spinal cord from adult mouse was dissected and dissociated by mechanical trituration.The tissue suspension was cultured in serum-free DMEM/F12 medium supplemented with EGF and B27.The cell colonies generated from a single cell were screened by limited dilution and incubated with BrdU.The cell colonies were transferred into medium with serum to induce differentiation.The cells were identified with antibodies to Nestin,BrdU,MAP2 and GFAP by immunofluorescence staining.Results The cells were cultured for seven days to generate proliferative neurospheres.The majority of cells in these neurospheres expressed Nestin and were differentiated into MAP2-positive cells and GFAP-positive cells in medium containing with fetal bovine serum.Conclusion A significant number of neural stem cells are present in the parenchymal adult mouse spinal cord and can proliferate and also give rise to neurons and glia in vitro.

Objective To compare MR,CT,and radiography in the detection of arthropathies in patients with hemophilia.Methods Forty-one symptomic joint images in the 14 patients with hemophilia, aged from 11 to 24 years,were used in this study.Each joint had the examinations of radiography,CT and MR within one day.The severity of each joint was staged using conventional radiographic classification. Severe HA patients with stage 5 were excluded from the study.Imaging findings of soft tissue swelling, osteoporosis,epiphyseal overgrowth,joint erosion,cyst,joint space narrowing,bone marrow,joint effusion, hemorrhage,synovial hypertrophy,widened intercondylar notch as well as anterior and posterior crueiate ligaments(only for knee joint)were used for the all imaging comparison.Results The 41 symptomatic joints in 14 patients with hemophilia were classified by radiographic criteria into stage 0(n=5),stage 1(n=7),stage 2(n=6),stage 3(n=8)and stage 4(n=15).Soft tissue swelling or joint effusion was observed in 33 joints by radiographs,in 34 joints by both CT and MR.Joint erosions were demonstrated in 34 joints by MR,in 33 joints by CT and 20 joints by radiographs.Joint cysts were shown in 21 joints by MR,in 18 joints by CT and 9 joints by radiographs.Significant differences in detection of erosion and cyst were found between radiography with either CT(P0.05).MR showed improvement for detecting nlore loci of both erosion and cyst than CT and radiography,and also CT showed the improvement than radiography.Bone marrow edema 14 joints, hemon'hage in 34 joints and synovial hypertrophy in 27 joints were revealed on MR images.Conclusion MRI is superior to CT and conventional radiography in detecting the abnormal changes and should be considered as the first choice among the imaging modafities in evaluating hemophilic arthropathies.

Neuroendocrine tumors (NETs) is a rare and heterogeneous group of tumors with widely varying morphologies and behaviors. Due to their rarity and heterogeneity, progress in improving its treatment has been slow. Pancreatic neuroendocrine tumors (pNETs) is a subset of NETs, previously known as islet cell tumors, occupies 3% of the primary pancreatic tumors with the annual incidence rate of (1-2)/100 000. In recent years, it is very necessary to improve the diagnosis and treatment of pNETs.

According to DNA sequences of the invA gene of Salmonella spp.,the phoA gene of Escherichia coli and the nuc gene of Staphylococcus aureus,three pairs of oligonucleotide primers were designed and synthesized to amplify the special DNA sequences by multiplex PCR. Moreover,the reaction conditions of multiplex PCR were optimized. The results showed the multiplex PCR using the three pairs of primers produced specific amplicons of expected sizes,284bp for Salmonella spp.,622bp for Escherichia coli,484bp for Staphylococcus aureus. The optimized reaction conditions followed as the concentration of primer 40nmol/L for Salmonella spp.,40nmol/L for Escherichia coli,80nmol/L for Staphylococcus aureus,2.4mmol/L Mg 2+ ,200?mol/L dNTP,1.5U Taq DNA polymerase,anneal temperature from 55.0℃ to 57.4℃. Under the condition,the detection limits for DNA template were 10.2pg,10.2pg and 102.0pg for Salmonella spp.,Escherichia coli and Staphylococcus aureus,respectively. The whole process could be completed within 4h. The multiplex PCR assay was a specific,sensitive,rapid and reliable method for detecting Salmonella spp.,Escherichia coli and Staphylococcus aureus,which establish important foundation for simultaneous detection for these three bacteria in food.

Objective To evaluate the roles of magnetic resonance imaging and proton magnetic resonance spectroscopy(1H-MRS) in the diagnosis of meningiomas. Methods 98 patients with meningiomas underwent conventional pre-contrast MR and contrast MR. Among them, 28 cases had two dimensional single voxel or multi voxel 1 H-MRS simultaneously both in the lesion's region and the contralateral side. Results On precontrast MR images of 98 cases, T1 WI showed 58.1% (61/105) isointensities, 31.4% (33/105) faintly low intensities and 10. 5% (11/105) mixed intensities; T2WI showed 40. 0% (42/105) isointensities, 41.0%(43/105) hyperintensities, 10.5% (11/105) faintly low intensities and 8.5% (9/105) mixed intensities. After administration of Gd-DTPA, the solid part of the tumors exhibited various enhancement in all the 98 cases.28 cases of MRS exhibited specific different spectral peaks, including increased of choline-containing compounds(Cho), absent or decreased of acetylaspartate(NAA), and the unchanged of creatine(Cr). The value of NAA, Cr, Cho, NAA/Cr, Cho/Cr, NAA/Cho in the tumor center of meningioma were 0. 09 ± 0.06,0.31 ± 0. 22, 0.46 ± 0. 16, 0.33 ± 0. 42, 1.50 ± 0. 68, 0. 15 ± 0.08, compared with the contralateral normal region, Cr has no significant difference (P ＞ 0. 05), NAA, Cho, NAA/Cr, Cho/Cr, NAA/Cho had significantly differences(P ＜ 0.05). Conclusion Conventional pre-contrast MR and contrast MR is the most important dignostic means for meningiomas, 1H-MRS combined with MRI can improve the diagnostic accuracy of meningiomas.

ObjectiveTo ascertain the safety and function of the transplantation of neonatal pig islets (NPIs) for diabetic patients.MethodsNPIs were injected into the hepatic artery of 22 patients.After the transplantation,the patients were treated with a multiple drug immunosuppressive regimens.The first 14 patients were treated with cyclosporine (CsA),mycophenolate mofetil (MMF) and prednisolon,and porcine C-peptide was not monitored,the following 2 patients were given cyklosporin and MMF only,while the next 6 patients were given a quadruple drug regimen consisting of OKT3,takrolimus,sirolimus and prednisolon.The blood glucose levels,exogenous insulin requirement,HbA1c,porcine endogenous retrovirus (PERV) and liver function were assessed before and after NPI transplantation.The serum porcine C peptide were monitored in last 8 patients.ResultsThe first 14 patients required less insulin and the HbAlc dropped after the transplantation.In the 2 subsequent patients,the metabolic parameters remained unchanged and monitor of porcine C-peptide was negative.Insulin requirements were reduced in all 6 patients,and HbAlc was normalized 3 months after the transplantation.Significant levels of porcine C-peptide were detected in the patient serum.Two of the patients were given a second injection of NPIs,and one of them became insulin independent for 7 d.No serious adverse events were noted after the transplantation.There was no evidence of PERV transmission.Six out of the 22 patients were followed up for 4-6 years after the NPIs injection,immunosuppressive treatment was stopped 1 year after the transplantation.The patients started to take insulin at the time of follow up.Four patients restricted the intake of sugar,while the other 2 did not.One patient had ketoacidosis twice and slight diabetic retinopathy,and another patient had ketoacidosis induced by acute gastroenteritis.The remaining 4 patients did not have any complications.Assays for PERV were again negative.ConclusionXenogenic islets can survive and function in the human body.No serious adverse events are noted.