Objective To investigate the indications,techniques and effects in surgical treatment of stale acetabular comminuted fractures.Methods A retrospective review was made on 24 cases (24 hips) of obsolete acetabular fractures undergone delayed reconstruction due to the combined injuries in other organs from October 2003 to June 2010.There were 18 males and 6 females,at 22-68 years of age [mean (42.2 ± 10.5) years].Mean time from injury to operation was (4.6 ± 1.3) weeks (range,3-6 weeks).According to Letournel classification,there were 11 simple fractures including six posterior wall fractures,one anterior wall fracture,one posterior column fracture,one anterior wall fracture and two transverse fractures and 13 compound fractures including six posterior column with posterior wall fractures,two transverse with posterior wall fractures,one T-shape fracture,two posterior semitransverse fractures and two both-column fractures.Surgery was performed via ilioinguinal approach in two cases,posterior acetabular approach in 13 and the combined approaches in nine.Results All 24 cases were available to average (15.6 ± 10.5) months of follow-up (range,10-36 months).Quality of reduction based on Matta standard was anatomic in 21 cases,unsatisfactory in two and poor in one.Clinical results evaluated using modified d' Aubigne-Postel score were excellent in 21 cases,good in two and poor in one.There were one femoral head necrosis,11 heterotopic ossification and six sciatic nerve transient palsy after operation.Conclusion Delayed open reduction and internal fixation is effective in treatment of obsolete acetabular comminuted fractures.

Objective To construct the genetic recombinant adenovirus vector carrying the human growth and differentiation fac-tor-5(GDF-5) gene by using AdEasy-1 adenovirus vector system and to amplify and prepare the recombinant adenovirus in HEK 293 cells .Methods Human GDF-5 gene obtained by PCR was inserted into plasmid pMD19-T and the 1 .7 kb GDF-5 cDNA sequence was cloned into the adenoviral shuttle plasmid pShuttle-cytomegalovirus(CMV) of the AdEasy-1 system .It was identified by DNA sequencing and a digestion with Hind Ⅲ restriction enzyme .The resultant pShuttle-CMV-GDF-5 was used to generate the adenovi-ral GDF-5 vector through homologous recombination with the adenoviral backbone plasmid ,pAdEasy-1 in BJ5183 bacterial cells .It was selected by kanamycin and identified by a digestion with Hind Ⅲ restriction enzyme and amplified in XL10-Gold competent bac-teria .The DNA of recombinant adenovirus vector was finally linearized by Pac Ⅰ and the adenoviral recombinants were used to pro-duce adenoviruses in HEK293 packaging cells ,resulting in an Ad-GDF-5 identified by Western blot .The virus titer was assayed by TCID50 .Results GDF-5 cDNA sequence obtained by PCR was 1 .7 kb .Gene sequencing results indicated that the sequence was i-dentical with the one in GENBANK .Cloned sequence 1 .7 kb(GDF-5) was obtained by a digestion with Hind Ⅲ restriction enzyme after GDF-5 cDNA segment was cloned into pShuttle-CMV and AdEasy-1 .Western blot showed that two bands migrating at ap-proximately 15 and 55 kDa were clearly observed in PVDF membrane .These data confirmed that HEK293 cells expressed a large number of mature GDF-5 protein after infected with Ad-GDF-5 .Our research results demonstrated that recombinant adenovirus vector GDF-5 was successfully constructed .The virus titer was 5 .6 × 109 PFU/mL .Conclusion Recombinant adenovirus vector carrying the human GDF-5 gene is successfully constructed by using the AdEasy-1 adenovirus vector system .Moreover ,the Ad-GDF-5 with high titer is prepared .These provide the basis for further study of the biological function of GDF-5 and the gene thera-py of its related diseases .

Objective To develop an MR optical dual-modality probe targeting angiogenesis of gastric cancer and to study its physical characteristics , in vitro cytotoxicity and magnetic effects of different pulse sequences on 3 T clinical MR scanner.Methods We conjugated GX1-Cy5.5, a novel gastric cancer neo-vasculature targeted peptide labeled with Cy 5.5, to the surface functionalized magnetic nanoparticles according to different molecular weights (1∶100, 1∶500),resulting in dual-modality probe DPs100 and DPs500 (named DPs).The hydrodynamic size and zeta potential of DPs and DPs 500 were analyzed by nano-ZS.The human umbilical vein endothelial cells (HUVECs) and BGC-823 cells were treated with DPs for 24 h, and methyl thiazol tetrazolium ( MTT) method was used to detect the survival rate of cells.DPs with different concentrations were scanned on different MR sequences , and then the relative signal intensity was observed.The absorbance of HUVECs and BGC823 cells treated with DPs of different concentration (0.00, 1.25, 2.50, 15.00, 50.00, 100.00 and 150.00 μg/ml) were compared with single factor analysis of variance.Relative signal intensity of different MR sequences was compared using a paired Wilcoxon signed-rank test.Results The dual-modality probe targeting angiogenesis of gastric cancer was successfully constructed.The hydrodynamic size of iron oxide nanoparticles , DPs100 and DPs500 was (35.23 ±0.07), (39.49 ±0.16) and (40.43 ±1.70) nm and the Zeta potential was (0.31 ±0.20), ( -4.15 ±0.79) and ( -10.51 ± 2.37) mV.The coupled rates of DPs 100 and DPs500 with polypeptide were 92%and 94% respectively.The absorbance of HUVECs and BGC823 cells treated with DPs of different concentrations were 0.76 ±0.04, 0.80 ±0.03, 0.79 ±0.05, 0.75 ±0.06, 0.74 ±0.05, 0.77 ±0.01,0.71 ±0.04 and 0.38 ±0.04, 0.43 ±0.04, 0.41 ±0.03, 0.43 ±0.07, 0.44 ±0.04, 0.41 ±0.07 and 0.40 ±0.04, there was no statistical significance ( F=0.94, 0.51;P>0.05).The signal intensity increased first and then decreased following the increasing concentrations of DPs on T 1WI,especially on FSPGR T1WI (Z =-3.294,P <0.05), while the signal intensity decreased on T2WI or T2*WI.There was no significant differences in signal intensity on FSE T2 WI and SSFSE T2*WI with iron concentration >10μg/ml( Z=-7.110,P>0.05).With iron concentration≤10 μg/ml,the signal intensity on SSFSE T 2*WI was significantly decreased compared to FSE T2 WI ( Z =-2.023, P <0.05 ) .Conclusions DPs may be potential dual-modal probes for characterization of tumor angiogenesis by MR and optical imaging noninvasively , without causing significant effects on the cell activity in vitro , and SSFSE T2*WI may be the most sensitive sequence for DPs evaluation on MR.

Objective To investigate abnormal protein expression of matrix metalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in human gastric adenoearcinoma, and further reveal the clinical significance. Method The MMP-9, VEGF and PCNA proteins expression was determined by immunohistochemistry staining in 45 gastric adenocarcinoma tissues, 45 adjacent specimens and 10 normal gastric mucosa tissues via tissue arrays accordingly. The relationship of these protein expression with differentiation degree, development and progression of gastric adenocarcinoma were also analyzed. Results Positive rates of MMP-9, VEGF and PCNA in gastric adenocarcinoma, adjacent specimens and gastric normal mucosa were as follows: MMP-9, 82.2%(37/45), 64.4% (29/45), 30.0% (3/10) (P=0.019); VEGF, 73.3% (33/45), 62.2% (28/45), 30. 0% (3/10) (P=0.029); PCNA,84.4% (38/45), 71.1% (32/45), 10.0% (1/10) ,there were statistically significant difference (P = 0. 001). The positive rates of MMP-9, VEGF and PCNA in well-differentiated adenocarcinoma, moderately differentiated adenocarcinoma and poorly differentiated adenocarcinoma were as follows: MMP-9,70.0%(7/10), 80. 0% (8/10), 88.0%(22/25), there were statistically significant difference (P=0.015);VEGF, 50.0%(5/10), 60.0% (6/10), 88.0% (22/25), there were statistically significant difference (P =0.000);PCNA, 60.0% (6/10) ,90.0% (9/10) ,92.0% (23/25) ,the difference is significant statistically (P = 0.004). The expression of MMP-9, VEGF and PCNA showed positive relationship with each other by rank correlation analysis (P < 0. 05). Conclusion Tissue arrays technology is effective tool to analyze the expression of cancer related proteins in gastric adenocarcinoma. The expression of MMP-9, VEGF and PCNA proteins participates in the tumorigenesis and development process of gastric adenocarcinoma, and these can be used as indexes to evaluate prognosis in clinical.

Super hydrophobic interface modified with silver nanoparticles was fabricated for the detection of pesticide residues.By using a chemical reduction method,silver nanoparticles were deposited on the substrate surfaces with different microscopic pore structures.Two kinds of composite substrates,including regular stainless steel mesh and cellulose polyester film,were used.The pre-treatment of the substrate with fluoridated reagents was used to form a super hydrophobic interface,which made the target molecules on the surface concentrate effectively.The surface with the cellulose polyester substrate was used to detect Rhodamine 6G (R 6G) effectively with surface enhanced Raman scattering (SERS) technique.The results showed that the detection hmit was 10-16 mol/L.In addition,the surfaces based on the stainless steel mesh and cellulose polyester substrate were used to detect trichlorfon pesticide with detection limits of 1 × 10-15 mol/L and 1 × 10-16 mol/L,respectively.

Objective To investigate the clinical efficacy of preoperative full reset combined with minimally invasive treatment of extreme distal pilon fractures.Methods A retrospective analysis was made on 34 patients (35 ankles) with tibial fractures extremely close to the distal articular surface treated surgically between January 2011 and January 2015.There were 21 mnales and 13 females,aged 20-71 years (mean,36.2 years).Injury resulted from traffic accidents in 32 patients and high falls in two.Using the AO/OTA fracture classification system,type 43-B3 was noted in three patients,43-C1 in five patients,43-C2 in 18 patients and 43-C3 in eight patients.Calcaneal traction combined with manipulative reduction was used to correct fracture displacement preoperatively.All fractures were stabilized by minimally invasive percutaneous plate osteosynthesis (MIPPO) through single or combined medial,anteromedial and anterolateral approaches while minimizing damage to bone attachment and continuity of soft tissue,after soft tissue swelling subsided.For the patients with articular surface collapsing with severe comminution,a series of procedures were done under direct vision including using the talus articular surface as a mold,stable fixation with fine Kirschner (1-1.5 mm) and thin screws (2.1-2.7 mm series) and impaction bone grafting below subchondral bone.Thereafter,distal tibia anatomical short multi-directional locking plate fixation,distal nail support and early ankle joint functional exercise were done.Burwell-Charnley radiological evaluation system was used for radiological assessment,and TeenyWiss scoring system for ankle clinical symptoms and function.Postoperative complications were recorded.Results Follow-up lasted for 11-38 months (mean,16.6 months).No infection,wound disunion,or plate exposure occurred.Burwell-Charnley radiological evaluation system showed anatomic reduction in 32 patients,unsatisfactory reduction in one,and poor reduction in one.According to the Teeny-Wiss scoring system,the results were excellent in 31 patients,good in two and poor in one,with the excellentgood rate of 97％.Three patients suffered traumatic arthritis after operation and alleviated after oral administration of painkiller.Conclusion With use of full reset combined with manipulative reduction to correct fracture displacement,minimally invasive locking plate,distal row of nails,impaction bone grafting and limited fixation,the patients with extremely distal tibial pilon fractures achieve satisfactory reduction,stable fixation,and early functional exercise.

Objective To explore the optimal conditions of preparing 68Ga-DOTA-iNGR (NGR peptide containing CendR motif),to evaluate its biodistribution in normal mice and to perform microPET imaging in tumor-bearing nude mice.Methods 68Ga fresh eluent (200 μl,92.5-129.5 MBq) obtaining with 68Ge-68Ga radionuclide generator was used to label DOTA-iNGR.The optimal conditions of labeling including pH,temperature,reacting time and concentration of DOTA-iNGR were determined.Then,the in vitro and in vivo stability and octanol/water partition coefficient of 68Ga-DOTA-iNGR were further analyzed.The biodistribution in normal Kunming mice was examined at 10,20,40,60 and 120 min after injection of 68Ga-DOTA-iNGR.Nude mice bearing HT-1080 (CDl3-positive) and HT-29 (CDl3-negative) tumors were established and underwent microPET imaging at 1 h after the intravenous injection of 68Ga-DOTA-iNGR.Data were analyzed using independent-sample t test.Results The optimal conditions of labeling was mixing 2 μg DOTA-iNGR peptide with 200 μl 68Ga (92.5-129.5 MBq) at pH 4.0,temperature 90-100 ℃ for 5-10 min.Under this condition,labeling rate reached (97.5± 1.3)％.The radiochemical purity of 68Ga-DOTA-iNGR in both saline (room temperature) and mouse serum (37 C) were both above 95％ after 4 h incubation,and the radiochemical purity in urine was greater than 85％ after 1 h metabolism in vivo.The partition coefficient was-2.71±0.18.In normal mice,majority of 68Ga-DOTA-iNGR was excreted from kidneys with a rapid clearance from blood.The in vivo microPET imaging showed that 68Ga-DOTA-iNGR was remarkably accumulated in the CD13-positive HT-1080 tumor.Conclusions Labeling DOTA-iNGR with 68Ga under our condition is a simple and efficient procedure with high labeling rate and high specificity.The product 68Ga-DOTA-iNGR has high stability,ideal biodistribution,and specific binding to CD13-positive tumor,which means that it's a very promising molecular probe for noninvasively detecting CD13-positive tumor.

Objective To evaluate the value of integrated use of bedside lung ultrasound and echo-cardiography in PICU.Methods Two cases with respiratory failure and shock were monitored using inte-grated bedside lung ultrasound and echocardiography.Breathing and circulation support solutions were adopt-ed according to Bedside Lung Ultrasound in Emergency(BLUE)and Fluid Administration Limited by Lung Sonography(FALLS).Results Case 1 with acute lymphoblastic leukemia complicated with severe phneu-monia was measured as “B-profile”throughout both lung fields which interpreted as pulmonary edema ac-cording to the BLUE protocol.He was diagnosed as acute respiratory distress syndrome later and supported by non-invasive ventilation according to the lung ultrasonography.After 4 days,the numbers of B line were sig-nificantly reduced with PaO2/FiO2 >300 mmHg(1 mmHg=0.133 kPa)and the improvement in chest X ra-diography was found,ventilation was weaned accordingly.Case 2 was diagnosed as septic shock and acute re-spiratory distress syndrome with volume resuscitation and mechanical ventilation.According to FALLS proto-col,we ruled out obstructive and cardiogenic shock,and assessed the variation in inferior vena cava diameter and aortic systolic velocity-time integral as a guide to fluid therapy.At the 8th hour in PICU,case 2 recovered from shock.Conclusion Integrated use of bedside lung ultrasound and echocardiography is clinically signifi-cant for the rescue of critically ill patient with respiratory failure and shock.

Objective TostudythevalueofquantitativeparametersofDCE-MRIandthreedimensionalarterialspinlabeling(3D-ASL) inpreoperativegliomagrading.Methods 70patientsdiagnosedpathologicallywithinitialgliomawereassessedretrospectively,including 32caseswithlow-gradeglioma(LGG)and38caseswithhigh-gradeglioma(HGG).Allpatientsunderwentconventional,enhanced, DCEand3D-ASL MRIat3.0Tbeforesurgery.TheparametricvaluesofDCEsuchasvolumetransferconstant(Ktrans),extravascular extracellularspacevolumefraction(Ve),therateconstant(Kep),fractionalplasmavolume(Vp),cerebralbloodflow (CBF)andcerebral bloodvolume(CBV)wereobtainedbycorrespondingpost-processingsoftware.ThecerebralbloodflowofASL (ASL-CBF)wasalso obtained.Ttestoftwoindependentsampleswasusedtoanalyzewhetherthemaximumandaveragevaluesofeachparameterwere statisticallydifferentbetweenLGGand HGG.Thediagnosticaccuracyofdifferenttechniqueforgliomagradingwasdeterminedby ROCcurveanalysis.Results ThemaximumvaluesofDCE-Ktrans,Ve,rCBVandmaximumvalueofASL-rCBFwerestatisticallydifferent betweentheHGGandLGG (P＜0.05).AlltheparametricaveragevalueswerestatisticallydifferentbetweentheHGGandLGG (P＜0.05).ThemaximumandaveragevaluesofKtranshadarelativelyhighestdiagnosticefficiencyinallparameters,withtheAUCwere0.986 and0.971,theoptimumthresholdwere0.264and0.068,thesensitivitywere93.3%and94.1%,andthespecificitywere100%and 100%,respectively.ThemaximumvaluesofVe,rCBV,ASL-rCBFandtheaveragevaluesofallparametershadarelativelyhigher diagnosticefficiency.Conclusion ThemaximumvaluesofKtrans,VeandrCBFofDCE,themaximumvalueofASL-rCBFandtheaverage valueofeachparameterwereusefultodistinguishbetweenLGGand HGG.ThemaximumandaveragevaluesofKtransarethebest parametersforidentifyingHGGandLGG.

Objective To investigate the methods of screening specific aptamers for (EpCAM)Gpositive prostate cancer (PCa)cells by cellGSELEX technique.Methods A random DNA library was designed to screen EpCAMGspecific DNA aptamers from human prostate cancer cells expressing EpCAM molecule by cellGSELEX technique.After 12 rounds of in vitro screening,DNA products were cloned and sequenced.Flow cytometry and cellular immunofluorescence were used to detect the specific binding ability of aptamers to target cells.Results Two aptamers of Ep1 and Ep2 were selected.Both of them could specifically bind to EpCAMGpositive cancer cells LNCap,PCG3 ,DU1 45 , and HEK293T cells transfected with target molecule.The binding rates of Ep1 were 61.0%,74.3%,5 9.1% and 60.3%.The binding rates of Ep2 were 65.1%,77.8%,54.2% and 58.3%.Neither of them could bind to HEK293T cells transfected with empty vector with the binding rate of 5.4% in Ep1 and 3.3% in Ep2,respectively.Flow cytometry analysis and confocal images indicated that the EpCAM aptamers could specifically recognize human PCa cells expressing EpCAM,but could not bind to EpCAMGnegative cells.Conclusion EpCAM aptamers derived from cellGSELEX technology can recognize and bind to EpCAMGpositive PCa cells specifically,which may provide new ideas for the specific diagnosis and targeted therapy of prostate cancer,and lay an experimental basis for the other specific diagnosis and treatment schemes of malignant tumors.