Objective: To construct and identify a lentiviral vector harboring RNAi sequence targeting the human high mobility group A1(HMGA1) gene.Methods: The effective sequence of siRNA targeting the HMGA1 gene confirmed in our previous study,the complementary DNA containing both sense and antisense Oligo DNA of the targeting sequence was designed,synthesized and cloned into the pGCL-GFP vector diced by the restriction enzyme of HpaⅠ and XhoⅠ,which contained the U6 promoter and green fluorescent protein(GFP).The resulting lentiviral vector containing HMGA1 shRNA was named LV-sh HMGA1 and confirmed by PCR and DNA sequencing.A total of 293T cells were cotransfected with LV-sh HMGA1,pHelper 1.0 and pHelper 2.0.All the virus stocks were produced by Lipofectamine2000-mediated transfection.The titer of the virus was tested according to the expression level of GFP.Results: PCR analysis and DNA sequencing demonstrated that the RNAi sequence targeting the human HMGA1 gene was successfully inserted into the lentiviral vector.The titer of the recombinant lentiviral vector was 5?107 TU/ml.Conclusion: The successful construction of the lentiviral vector of HMGA1 has prepared the ground for further studies on the functions of the HMGA1 gene with the RNAi technique.

Objective To further study the value of MR T2WI fat saturation sequence in acute Spinal Trauma. Methods 109 cases of acute Spinal Trauma underwent MR imaging were analysed retrospectively. The detection rate of lesions with bone marrow edema, Vertebral fracture and spinal cord injury was compared in T2WI and FS T2WI. Results FS T2WI could detect more lesions of obscure fracture and bone marrow edema than that in T2WI,and it could accurately show the extent and the feature of anatomic structure in acute injury of bone. Conclusion FS T2WI can improve the diagnostic sensition and accuracy in Spinal Trauma, and it should be used as a routine MR sequence for detecting Spinal Trauma.

Objective To investigate the clinical and angiographic characteristics of no reflow phenomenon after primary percutaneous coronary intervention (PCI) with acute myocardial infarction (AMI).Methods A total of 319 patients with AMI undergoing primary-PCI was divided into no-reflow and normal reflow groups.The incidence of no-reflow phenomenon,the clinical date,angiography findings,and surgical date were compared between two groups.Results No-reflow phenomenon occurred in forty(13.4％)of the patients after primary PCI.There was dramatic difference in combined hyperlipidemia,angina pectoris history before AMI,heart function ≥2 grades on admission,the length of the vascular lesions,vascular stenosis degree,blood clot load level,coronary artery opening time,and the expansion of the balloon between no-reflow and normal blood flow groups.Multiple logistic regression analysis identified that angina pectoris history before AMI,heart function classification on admission,high thrombus burden,the expansion of the balloon,and coronary artery opening time on angiography as independent predictors of no-reflow phenomenon.Conclusions The occurrence of no-reflow phenomenon after primary PCI was associated with high cholesterol history,no history of pre-infarction angina,heart function classification on admission,long vascular lesions,narrow degree of heavy,blood clots in the high load,coronary artery opened long time,and the expansion of the balloon more frequently.

Objective To investigate the effect of Cimetidine on the adhesion between colorectal cancer LOVO cells and endothelial ECV304 cells;and study whether Cimetidine can inhibit the expression of E-selectin in ECV304 cells. Methods Cellular uptake of rose Bengal stain was used to measure the adhesion between LOVO cells and ECV304 cells. Enzyme-linked immunosorbent assay(ELISA)and flow cytometry (FCM, indirect fluorescence staining and labeling)were used to detect the expression of E-selectin. Results ECV304 cells were exposed to different concentrations of Cimetidine. Both the adhesion between LOVO cells and ECV304 cells and the levels of E-selectin significantly decreased with increasing concentration of Cimetidine(P =0. 001 and 0. 001 respectively). Conclusion Cimetidine can inhibit the adhesion of human colorectal cancer LOVO cells on endothelial ECV304 cells by blocking E-selectin expression. Our observations indicated a potential of anti-adhesion therapy using Cimetidine in cancer treatment.

Objective According to the characteristics of skin defects in ankle and foot select the posterior lateral leg pedide skin flaps supplyed by different blood to repair,providing advice and reference.Methods Between January,2001 and December,2015,163 cases of soft tissue defects at the foot and ankle were treated in our department.①The sural neurovascular flap was used in 93 cases in ankle and foot defect.②The sural nerve nutritional vessel flap pediele with the perforating branch of the peroneal artery in 36 cases in ankle and foot defect.③The sural nerve nutritional vessel flap pediele with the perforating branch of the peroneal artery including sensory reconstruction in 16 cases in heel rejion defect.④Peroneal artery perforator flap in 10 cases in anterior ankle,lateral malleolus and posterior malleolus defect.⑤The use of the sural fasciocutaneous flap alonged with peroneal artery and perforators in 8 cases in dorsal foot defect.The donor site with skin graft.Results Of the 93 sural neurovascular flaps,8 had partial loss,which were cured after dressing.All the sural nerve nutritional vessel flaps pediele with the perforating branch of the peroneal artery survived.Sensory grading standard by UK Medical Research Council was used to evaluate the recovery of sensory function on the last follow-up.The sensory function recovery of heel region flaps with reconstruction of the sensory was between S0 and S1.All peroneal artery perforator flaps survived.One of the sural fasciocutaneous flap alonged with peroneal artery and perforators occurred distal epidermis,which were cured after dressing.All patients were followed up 6-50 months (mean 20 months).All patients had recoveryed walking function,and infection wound had no recurrence after surgery.Conclusion According to the location,size,severity and the injury of peripheral vascular,select the most simple,safe,minimal damage flap for the soft tissue defects at the foot and ankle.The right choice and the exact design can improve flap survival rate,and recieve good clinical results.

A poly( GMA-EGDMA) coated SPME fiber was prepared using an in-situ polymerization by direct bonding to the surface of a polydopamine-modified stainless steel wire. Then the fiber was modified by sulfuric acid. A novel solid phase microextraction coating coupled to high performance liquid chromatography ( HPLC) method based on the as-prepared fiber was developed for the determination of four pharmaceuticals and personal care products ( PPCPs) in water samples. The influences of extraction parameters, including pH, extraction time, extraction temperature and salt addition were investigated. 3 mL water sample was extracted by the as-prepared fiber for 60 min at 30 ℃, and then desorbed with mobile phase for 30 min, respectively. Desorption solution was analyzed by HPLC-DAD ( diode array detection ) . The results indicated that the extraction yield of the fiber was good for four PPCPs. The linear correlation coefficients were>0. 997 with the linear range of 2-200 μg/L. The limits of detection (S/N=3) were 0. 5-5 μg/L with RSD (n=5) of 4. 1%-11. 9%. The recoveries of four PPCPs at spiked level of 20, 50, 100 μg/L were within the range of 70. 6%-105. 5%. The results showed that this method was easy, green, accurate and precise, and could be used to assay the four PPCPs in real water samples.

Objective:To find absorbable adhesives with suitable bonding properties for the absorbable polylactic acid root canal post. To test and compare the bond strengths of absorbable polylactic acid root canal post with three different adhesives. Methods:The absorbable polylactic acid root canal posts were used to restore the extracted teeth, using 3 different adhesives: cyanoacrylates, fibrin sealant and glass ionomer cement. The teeth were prepared into slices for micro-push-out test. The bond strength was statistically analyzed using ANOVA. The specimens were examined using microscope and the failure mode was divided into four categories:cohesive failure between absorbable polylactic acid root canal posts and adhesives, cohesive failure between dentin and adhesives, failure within the adhesives and failure within the absorbable polylactic acid root canal posts. Results:The bond strength of cyanoacrylates [(16. 83 ± 6. 97) MPa] and glass ionomer cement [(12. 10 ± 5. 09) MPa] were significantly higher than fibrin sealant [ ( 1 . 17 ± 0 . 50 ) MPa ] , P <0 . 001 . There was no significant difference between cyanoacrylates and glass ionomer cement (P =0. 156). In the group of cyanoacrylates, the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives was 25 . 0%, the cohe-sive failure between the dentin and the adhesives was 16. 7%, the failure within the adhesives was 33. 3%, and the failure within the absorbable polylactic acid root canal posts was 25 . 0%. In the group of fibrin sealant , the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives was 66 . 7%, the cohesive failure between the dentin and the adhesives was 22 . 2%, the failure within the ad-hesives was 11. 1%. In the group of glass ionomer cement, the cohesive failure between the absorbablepolylactic acid root canal posts and the adhesives was 87. 5%, the failure within the adhesives was 12. 5%. The major failure mode in fibrin sealant and glass ionomer cement was the cohesive failure between the absorbable polylactic acid root canal posts and the adhesives. No major failure modes were found in the group of cyanoacrylates. Conclusion:The bond strength of fibrin sealant is low, which cannot meet the requirement of clinical use. The bond strengths of cyanoacrylates and glass ionomer cement are suitable for clinical use. The cyanoacrylates are a kind of absorbable adhesive which has suitable bonding proper-ties for the absorbable polylactic acid root canal post.

Objective To study the therapeutic effect of the complex mixture of luteolin and rutin ( MLR) on 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridin ( MPTP) induced Parkinson’ s disease ( PD) mouse model. Methods Seventy-two C57BL/6 mice were divided into six groups randomly ( n=12 in each group): the normal control , model control , madopar (50 mg·kg-1) group, MLR at low (140 mg·kg-1), middle (280 mg·kg-1) and high (560 mg·kg-1) dose groups. PD mouse models were established by intraperitoneal injection of MPTP ( 30 mg · kg-1 ) . Pole test and traction performance were recorded to access the body coordinate capability and strength. The tyrosine hydroxylase (TH), dopamine transport protein ( DAT) , and glial fibrillary acidic protein ( GFAP ) positive cells were detected by immunohistochemical method. Dopamine ( DA ) , dihydroxyphenylacetic acid ( DOPAC ) , homovanilic acid ( HVA ) , 5-hydroxytryptamine ( 5-HT ) and 5-hydroxyindoleacetic acid (5-HIAA) in striatum were quantified by HPLC-ECD. Results MLR significantly ameliorated mouse motor coordination ability (P<0. 01 or P<0. 05). MLR at 280 and 560 mg·kg-1 could increase TH-positive neurons by 69. 00%and 77. 95% compared with the normal control group (P<0. 01) and DAT-positive neurons by 68. 53% and 70. 40% compared with the normal control group(P<0. 05), and decrease GFAP-postive astrocyte reactivity. The treatment with MLR at three doses attenuated the monoamine neurotransmitter disorder. Conclusion MLR markedly improves MPTP caused movement coordinate ability injury in mice and exerts therapeutic action on PD by regulating neurotransmitters in brain, inhibiting the inflammatory reaction and alleviating the neuron injury.

BACKGROUND:Pedicle screw is the major instrumentation of surgery in thoracic spine. However, there have been few reports about pedicle morphology relevant to screw insertion tracts, and few reports comparing the normal adolescents and adolescent idiopathic scoliosis patients. OBJECTIVE:To compare the morphologic characteristics of the thoracic pedicle with regard to safe thoracic pedicle screw placement in normal adolescents and adolescent idiopathic scoliosis patients. METHODS: Thoracic pedicles of thirty-five normal adolescents and thirty-five adolescent idiopathic scoliosis patients were measured with three-dimensional reconstruction CT images. Measured parameters include (1) critical distance: the shortest distance from an entry point to the ventral cortex of the lamina. (2) Safe distance: the distance from the entry point to the tangent of the spinal canal at the medial wal of the pedicle. (3) Pedicle screw length. (4) Pedicle width. (5) Pedicle transverse angle. The dangerous area was defined as the distance between the critical distance and the safe distance. RESULTS AND CONCLUSION: The mean critical distance was (9.2±1.0) mm for the normal adolescents, and (9.4±1.2) mm for the adolescent idiopathic scoliosis patients. Safe distances were significantly less in normal adolescents (14.7±0.8) mm than that of the adolescent idiopathic scoliosis group (15.4±1.4) mm (P < 0.001). The dangerous area was (5.4±0.7) mm for the normal adolescents, which was significantly less than that of the adolescent idiopathic scoliosis patients (6.0±1.0) mm (P < 0.001). Pedicle screw length was (36.6±4.1) mm for the normal adolescents and (37.1±5.3) mm for the adolescent idiopathic scoliosis patients. Pedicle width was (5.8±1.2) mm for the normal adolescents and (5.7±1.7) mm for the adolescent idiopathic scoliosis patients. No significant difference in critical distance, pedicle screw length and pedicle width was found between the two groups (P=0.382, 0.135, 0.293). Pedicle transverse angle decreased gradualy from T1 to T12 in both groups. These results verify that pedicle morphology of many parameters is different between normal adolescents and adolescent idiopathic scoliosis patients, especialy in the apical area of the thoracic curve.

OBJECTIVE:To establish a method for the determination of related substances in bisacodyl raw material and enteric-coated tablet. METHODS:HPLC was performed on the column of Hibar C18 with mobile phase of acetonitrile-20 mmol/L ammonium acetate (acetic acid adjust pH to 5.0)(55∶45,V/V),detection wavelength was 265 nm,flow rate was 1.0 ml/min, column temperature was 30℃,and the injection volume was 20 μl. RESULTS:The linear range of bisacodyl was 0.25-5.0 mg/ml (r=0.999 9);the limits of detection and quantification were 19-25 ng and 61-68 ng for bisacodyl and impurity A,B,C,D and E;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 99.50%-101.00%(RSD=0.5%,n=9). CONCLUSIONS:The method is specific, sensitive and reproducible, and can be used for the determination of related substance in bisacodyl raw material and enteric-coated tablet.