Objective To construct specifically expressed vascular endothelial growth factor (VEGF) 165 gene in retina. Methods Rho promoter, specifically expressed in retina, was amplified by polymerase chain reaction (PCR) from the genomic DNA of a BLAB/C rat, then it was cut with restriction enzymes and cloned into the plasmid pcDNA 3.1+-VEGF 165 to form recombinant plasmid pcDNA 3.1+-rho-VEGF 165. The correct recombinant plasmid pcDNA 3.1+-rho-VEGF 165 was identified by restriction enzymes and PCR, and was transferred by jetPEI into cultured human navel vein endothelial cells and human retinal pigment epithelial (RPE) cells. The expression of VEGF protein in human navel vein endothelial and RPE cells was detected by immunocytochemical staining and protraction of the growth curve of the cells. Results In human RPE cells, the expression of VEGF protein was more in recombinant plasmid pcDNA 3.1+-rho-VEGF 165 than that in plasmid pcDNA 3.1+-VEGF 165; in human navel vein endothelial cells, no obvious difference of the expression of VEGF protein between recombinant plasmid pcDNA 3.1+-rho-VEGF 165 and plasmid pcDNA 3.1+-VEGF 165 was found. Conclusions The construction of pcDNA 3.1+-rho-VEGF 165 carrier may provide the basic material for the study of the nosogenesis of VEGF in retinal neovascularization, and establish the foundation to set up the model of transgenic mice with VEGF specific expressing in retina.

Objective:To develop and apply a method for real-time quantifying IL-6 mRNA.Methods:By Ficoll-Hypaque density gradient centrifugation, human peripheral blood mononuclear cells(PBMC) were isolated from whole blood treated with ethylenediaminetetraacetic acid(EDTA). Total RNA extracted from the PBMC in trizol solution was reverse transcribed to cDNA, which used as templates for fluorecent real-time quantitative PCR amplification of IL-6. PCR system consists of IL-6 primer, SBY Green Ⅰ and so on.Results:The method is wide linear range, sensitive, specific, reproducible and simple.Conclusion:The method can accurately quantify IL-6 mRNA.

Objective To assess the clinical manifestation, management and outcome of abdominal aortic aneurysm (AAA) in young adult Methods Retrospective study was made on young AAA patients (

Objective To construct a new recombinant immunotoxin expression vector fused with a murine interleukin18(IL18) gene and a truncated pseudomonas exotoxin (PE38) gene, and examine the expression of IL-18-PE38 fusion protein in Escherichia coli (E. coli). Method Murine IL-18 (mIL-18) cDNA was cloned from murine liver tissue through reverse transcription-polymerase chain reaction (RT-PCR). The mIL-18 cDNA was ligased with a PE38 gene carried by PRKL expression vector through T4 DNA ligase and constructed into fusion protein expression plasmid PRKL-IL18-PE38. The recombinant vector was identified by restriction endonucleases digestion, PCR and DNA sequencing. After transformed into E.coli BL21 and induced by IPTG, the expressed product was obtained and the molecular weight and specificity were determined by SDS-PAGE and Western-blotting. Result The new recombinant immunotoxin expression vector was constructed successfully. DNA sequencing revealed that the mIL-18 and PE38 gene were consistent with NCBI Gene Bank. The IL-18-PE38 fusion protein was expressed in E.coli BL21, and Western-blotting analysis indicated that the molecular weight of the expression product is about 56 kDa, and could react with the specific antibody against mIL-18. Conclusion IL-18-PE38 recombinant immunotoxin expression vector will provide the basis for study on the targeted cytotoxic activity to Th1 cells and may have some potential value in the treatment of Th1 cell-mediated autoimmune diseases.

ObjectiveTo study change of aminoacid content in the brain of newborn rabbits with bilirubin encephalopathy. MethodsNewborn rabbits (3-5 days after birth) with bilirubin encephalopathy were induced by administration intra peritoneally with bilirubin (200mg/kg). Aminoacid content in brain were analysed by Hitachi 835 50 type aminoacid analysis instrument.ResultsThe content of glutamate and aspartate in brain tissue with bilirubin encephopathy decrease; GABA, alanine, tyrosine, valine and pheny lalanine increase. ConclusionsThe change of content of neurotransmitters in brain indicates that excitotoxicity of EAAs may contribute to the bilirubin encephalopathy. The increase of non neurotransmitters level proves that bilirubin may inhibit the biologic composition of protein in brain.

ObjectiveTo explore the diagnostic methods and surgery pattern at the first time of spontaneous perforation of congenital choledochal cyst.MethodsEleven cases(4 male,7 female) with spontaneous perforation of congenital choledochal cyst were 6 months to 5 years old,and their average course of disease were 4 days.Gustily abdominal distension,abdominal pain,crying and fever were present in all cases.Jaundice(7 cases) and emesis(5 cases) appeared.All cases were detected with physical sign of peritonitis by physical examination.Choledochal cysts were confirmed by CT or B ultrasound in 8 cases.All cases accepted abdominal paracentesis and biliary ascites was drawn.Three different operative procedures were performed:choledochocyst excision & Roux-Y choledocho-jejunostomy(2 cases),choledochotomy with T-tube drainage(3 cases),and cholecystostomy(6 cases).Nine children receiving external drainage operation accepted a second operation to rebuild biliary tract(choledochocyst excision & Roux-Y choledochoje-junostomy) after 3 to 6 months.ResultsAll cases had got satisfactory therapeutic efficacy without any grave complication such as fistula of anastomotic stoma,infection of biliary tract or obstruction of biliary tract.During operation,perforations were located in the juncture of choledochus and cystic duct in 5 children and were not found in the other 6 children.In the second operation,the cases receiving cholecystostomy had less peritoneal adhesion,anatomic structure changes,haemorrhage[(30-50) mL vs(100-200) mL] and operation time[(2.5-3.0) h vs(3.5-5.0) h] than those receiving choledochotomy with T-tube drainage,and did not appear inadequate drainage for cystic duct obstruction.ConclusionsFor children with more organ inflammatory edema and adherence and in a bad overall condition,the first-time operation of cholecystostomy is more reasonable.

Objective To explore the effects of amisulpride on the cognitive function in first-episode schizophrenia patients.Methods 64 patients in first-episode schizophrenia were divided into two groups randomly according to the method of tossing a coin,then treated with amisulpride or risperidone respectively for 12 weeks.The efficacy and adverse effect were evaluated with positive and negative scale (PANSS) and treatment emergent side effect scale (TESS) before treatment and after treatment for 2 weeks,4 weeks,8 weeks,12 weeks.Before and after treatment for 12 weeks,cognitive function of all the patients was blindly evaluated with Wechsler Scale-revised China (WMS-RC),Wisconsin Card Sorting Test(WCST) and Trail Making test A and B.Results After 12-week treatment,statistical difference was found in amisulpride(44.7 ± 6.7) and risperidone (45.2 ± 7.4) groups (P ＜ 0.01).But no statistical difference was found between the two groups (P ＞ 0.05).The cognitive function in firstepisode schizophrenia was damaged obviously.In two groups,the scores in recognize,association,comprehend,back a few and MQ of WMS and TAT-A,TAT-B were improved significantly after treatment for 12 weeks (P ＞ 0.05).These items of WCST were improved more remarkably than baseline (amisulpride (20.63 ± 13.06),(28.75 ± 15.72),(43.17 ±22.13),(3.62 ±2.21),P＜0.05; risperidone(20.41 ±13.82),(29.31 ± 16.12),(42.78 ± 21.42),(3.67 ± 2.32),P ＜ 0.05).The improvement in the scores of WCST were statistical difference compared with control group(P ＜ 0.05).But statistical difference was no found between the two groups (P ＞ 0.05).Conclusion The study shows that the cognitive dysfunction in first-episode schizophrenia can be improved by amisulpride,and the efficacy was similar with risperidone.

BACKGROUND:The interaction of neural network and motor function in post-stroke brain tissue remains unclear.
OBJECTIVE:To observe neural network impairment fol owing subacute stroke by using diffusion tensor imaging, and to investigate the relationship with neurological defects and motor dysfunction.
METHODS:A total of 19 patients after subactue stroke and 20 healthy adults were examined with diffusion tensor imaging. The fol owing parameters were compared:fractional anisotropy, apparent diffusion coefficient, asymmetry indices of fractional anisotropy and apparent diffusion coefficient. The neurological defect and motor function were evaluated with the corresponding scales. The 10-meter walking speed was measured. The correlation of diffusion tensor imaging parameters with the scale scores and 10-meter walking speed was analyzed.
RESULTS AND CONCLUSION:The stroke group exhibited reduced fractional anisotropy value asymmetry and fractional anisotropy value in bilateral posterior limbs of the internal capsule. Apparent diffusion coefficient value asymmetry and apparent diffusion coefficient value in the posterior limb of the internal capsule were lower than control unaffected side (P<0.05). Apparent diffusion coefficient value and apparent diffusion coefficient value asymmetry in posterior limb of the internal capsule showed a strong negative correlation with Fugl-Meyer assessment scores of the lower extremities (P<0.05). Diffusion tensor imaging parameters is closely linked with motor dysfunction of the lower extremities in subacute stroke patients. Local stroke lesion-caused neurological defect is the leading cause of motor dysfunction of the lower extremities.

Aim To observe the apoptosis of human leukemic cells induced by F951, a novel bcl-2 antisense oligodeoxynucleotide. Methods HL60 cells were cultured with F951 in variant doses. Apoptotic cells were detected by flow cytometry, DNA ladder, electron microscope observation and terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL). Results After HL60 cells were treated for 48 hours, mitochondria apoptotic cells can be detected at the ratio of 3.00% in the untreated group, 13.57% in the FNS control group and 30.95%, 38.08%, 52.55% with 5, 10, 20 ?mol?L-1 F951 respectively; the ratio of cells with caspase activity was 0.08% in the untreated group, 0.14% in the FNS control group and 43.68%, 60.54%, 80.37% with 5, 10, 20 ?mol?L-1 F951 respectively. Typical DNA ladder was seen from gel electrophoresis in F951 treated groups, and more apparent effect in the aspect of inducing DNA ladder had been observed with the improvement of F951 concentration. Detected through TUNEL and electron microscope, apoptotic cells in untreated group and FNS control group can only be found by chance, but very commonly seen in F951 treated groups and more frequently with the improvement of F951 concentration. Conclusion F951 can induce cell apoptosis in HL60 cells. Such effect is achieved through the inhibition of bcl-2 gene expression by F951 which initialize apoptosis passageway in consequence.

Objective:To analyze the magnetic resonance imaging (MRI) features of parasellar cavernous haemangioma and improve the diagnosis and differential diagnosis accuracy.Methods:13 patients with parasellar cavernous haemangioma were collected.All the patients were diagnosed by MRI and confirmed by pathology.Based on the pathologic findings,the MRI features were discussed.Results:9 cases presented horizontal dumbbell.The lesions located in the parasellar were larger than the sella turcica.The main body position of the lesions were centered lateral to the parasellar and encasesed the intracavernous internal carotid artery (ICICA).While,1 cases were similar in size and shape.1 case was located in the sella turcica.2 cases was centered lateral to the ICICA.pituitary were detected obscurity in 7 cases and displaced in 6 cases,6 cases appeared extremely high homogeneous intensity on T2-weighted images:as bright as cerebrospinal fluid signal.Only 5 cases underwent three-dimensional arterial spin labelling perfusion imaging (3D-ASL).The lesions revealed marked hypoperfusion.The cases of misdiagnosis were 9,including 4 cpituitary adenomas and 5 meningiomas.Conclusions:The characteristics of MR images ofparasellar cavernous haemangioma were horizontal bottle gourd form,the main body position of the lesions were centered lateral to the parasellar and encasesed the (ICICA),and their extremely high homogeneous intensity on T2-weighted images:as bright as cerebrospinal fluid signal.In cases that are equivocal,3D-ASL were useful in differentiating cavernous haemangiomas from parasellar meningiomas,which could decrease mistaken diagnosis.Graspping the imaging feature and differential diagnosis were helpful for the diagnosis of this disease.