Endostatin is a potent, endogenous inhibitor of angiogenesis, which corresponds to the C-terminal fragment of collagen ⅩⅧ. The human endostatin gene was amplified from a human fetal liver cDNA library by means of PCR and was cloned into pPIC9K vector. Soluble endostatin was superexpressed in Pichia pastoris (50.5mg/L) . The recombinant protein was purified by cation exchange chromatography with the final yield of more than 95%. Western blotting analysis showed positive immunoreactivity to the expressed product. Recombinant endostatin was able to suppress the angiogenesis on the CAMs. Also, the endostatin inhibited specifically of the migration of HMECs stimulated by bFGF, with EC50 being about 0.4 ?g/ml.

Objective To explore the effects of gap junctional(GJ)proteins in pathogenesis of cerebral schistosomiasis, through observing the expression of gap junctional proteins Cx37 mRNA in cultured cerebral arterial endothelium incubated with soluble eggs antigen(SEA). MethodsCerebral artery endothelial cells of rabbits were incubated with SEA, and the experiments were divided into control group and SEA 1 - 5 groups (SEA concentrations were 10.0％ ,5.0％ ,3.3％ ,2.5％,2.0％, respectively), reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to examine the expression of Cx37 mRNA and protein in cerebral artery endothelial cells of rabbits, respectively. Results Cx37 mRNA levels of control and SEA 1 - 5 groups were 0.239 ± 0.037, 0.260 ± 0.043, 0.218 ± 0.310, 0.647 ± 0.040, 0.419 ± 0.036, and 0.513 ± 0.038, respectively;SEA 3 - 5 groups were higher than control group of mRNA levels(all P＜ 0.05). Cx37 protein levels of control and SEA 1 - 5 group were 0.401 ± 0.045, 0.485 ± 0.048, 0.749 ± 0.052, 1.119 ± 0.063, 1.015 ± 0.057 and 0.605 ±0.047, respectively, of which SEA 2 - 5 groups were higher than control group(all P ＜ 0.05). ConclusionsExpression levels of Cx37 mRNA and protein in cultured cerebral artery endothelial cells incubated with SEA are higher than those of control cerebral artery endothelial cells, which suggests that the gap junction proteins may play an important role in pathogenesis of cerebral schistosomiasis through SEA and its secretion in infiltration of brain tissue and deposition in the cerebral arteries.

Objective To evaluate the feasibility of monitoring the gene expression of VEGF165 via the diglycylcysteine (GGC) reporter gene system by reporter probe of 99Tcm-GH. Methods DNA fragments encoding GGC binding motifs were prepared by PCR and positioned at the C end of VEGF165 gene after the linearization of pcDNA3-VEGF165 plasmid. A replication-defective adenovirus vector Ad5-VEGF165GGC motif-internal ribosomal entry site(IRES) -enhanced green fluorescent protein (EGFP) (Ad5-VIE)was constructed, with a cytomegalovirus (CMV) early promoter driving the expression of VEGF165 gene,GGC motif and EGFP, under the aid of an IRFS. A replication-defective adenovirus carrying the Ad5-EGFP was used as the control. Mensenchymal stem cells (MSC) were infected with the recombinant adenovirus at a multiplicity of infection (MOI) from 0 to 100 infectious units (0,10,25,50,100). The cellular uptake of 99Tcm-GH in infected MSC were then studied at 30, 60, 90 and 120 min. VEGF165 was detected by quantitative reverse transcriptase real-time PCR (RT-PCR), Western-blot, and immunohistochemisty. EGFP was observed by RT-PCR and fluorescence microscopy. The correlation analysis was studied between the cellular uptake of 99Tcm-GH and the expression of VEGF165. SPSS 13.0 was applied for statistical analysis. Independent samples t-test, q-test and Pearson correlation coefficient were used. Results After infected with different viral titer of Ad-VIE, the cellular uptake of 99Tcm-GH increased with the increasing virus titer(r2 =0.86, P ＜0.05), with the peak rate (7.94 ±0.75) % at MOI = 100. In time-dependent uptake study, the cellular uptake rates increased rapidly with the time extension, and the highest uptake occurred at 120 min with the peak uptake rate (7.72 ±0.22)%. The uptake rates of 99Tcm-GH in Ad5-VIE-infected cells were significantly higher than those of Ad5 -EGFP-transfected cells at all time points (t = 15.10- 54.92, all P ＜0.05). The VEGF165 and EGFP mRNA levels increased with increasing virus titer, and the VEGF165 mRNA correlated well with the EGFP mRNA(r2 = 0. 99, P ＜ 0.05). After infected with different MOI of Ad5-VIE, good relationship was found between the cellular uptake of 99Tcm-GH and the expression of VEGF165protein in MSC(r2 =0.90, P ＜0.05). Inmunohistochemisty showed VEGF165 protein expressed obviously at Ad5-VIE-infected MSC, and the EGFP was observed by fluorescence microscopy. Conclusions The cellular uptake of 99Tcm-GH in Ad5-VIE-infected MSC are well correlated with the expression of VEGF165 in vitro. The expression of therapeutic gene VEGF165 can be monitored by the GGC peptide expression.

Background and purpose:It was reported that chemokine receptor CXCR4 and its ligand stromal cell-derived factor 1(SDF-1)were involved in the proliferation,differentiation,and metastasis of tumor.This study was designed to observe the expression of chemokine receptor CXCR4 in hypopharyngeal squamous cell carcinoma tissue and study the relationship between the expression of chemokine receptor CXCR4 and different clinicopathlogical characteristics,and further to explore the clinical significance.Methods:For the detection of the expression of chemokine receptor CXCR4,43 primary hypopharyngeal squamous cell carcinoma tissues,27 normal hypopharyngeal tissues,34 lymph node metastastatic lesions and 9 normal lymph node lesions were detected by immunohistochemical method using rabbit anti-human CXCR4 polyclonal antibody.Results:The positive expression rates of CXCR4 in 43 hypopharyngeal carcinoma tissues and normal tissues were 95.3% and 22.2%,respectively(P

Objective:To value the application of two bacteria colony-PCR methods in the screening of phage antibody library. Methods:Five positive monoclonal bacterium were respectively suspended in either deionized water or 0.1% Triton X-100, and then boiled to be used as template in PCR. . The DNAs products of PCR were extracted and digested by two enzymes, and then determined by electrophoresis. Results:The inserted genes were detected in all the 5 clones after PCR and enzyme digestion .Conclusion:Bacteria colony-PCR can be used in screening positive recombinant colonies. The bacteria colony-PCR method with bacteria colonies suspended in deionized water is valuable in large scale positive recombinant bacterium screening.

Aiming at molybdate and boron deficient acid purple soil from main peanut cultivated areas in Sichuan,and Mo and B requirement of peanut growth,the feasibility of Co-inoculum of peanut Bradyrhizobium and molybdate and boron was studied.The tolerance to molybdate and boron of the tested strains Spr2-9,Spr4-5 was inspected.The result indicated that the two tested strains could tolerate higher concentration of molybdate than that of boron.The compound inoculum of Bradyrhizobial strain and trace element Mo was developed.The optimum concentration of Mo was 0.4%.

Objective To discuss the frequent etiology of spontaneous sub-cortical hemorrhage and its diag- nosis.Methods The clinical materials of 79 cases of spontaneous sub-cortical hemorrhage were analyzed.Results 56% of the hemorrhage was caused by arterial-venous malformation.48% of the hemorrhage was caused by occult AVM.Conclusion AVM is the most frequent etiology of spontaneous sub-cortical hemorrhage.CTA plays a special role in its diagnosis.

Objective To investigate and analyze the clinical and imageological features of skull base erosion in nasopharyngeal carcinoma( NPC). Methods 67 NPC patients proved by pathology were retrospectively reviewed. All patients underwent both CT and MRI plain plus enhancement scan. Scan slices were performed from oral pharynx (lower border of second cervical vertebra) to suprasellar cistern by axial CT scans and axial, sagittal and coronal MRI scans. Results (1) 52 patients with skull base erosion were found with MRI, and only 35 patients with CT. The following structures and figures were difined with CT and MRI respectively: pterygoid plates 8,14; clivus 22,25 ; petrous apex 25 ,28 ; sphenoid body or sinus 19,25; sphenoid wing 9, 12; cavernous sinus 14,17. The display difference between CT and MRI had statistical significance ( x2= 9. 47 , P = 0. 02). (2) CT- defined skull base erosion was most bone destruction , however, not only bone destruction but also tumor crossing skull base structure could be defined by MRI. (3) The incidence of headache was 82.7% (43/52) , cranial nerve palsy was 67. 3% (35/52) , both headache and cranial nerve palsy was 57. 7% (30/52). Conclusion Headache and cranial nerve palsy are primary characteristics of the skull base erosion in NPC. MRI is superior to CT in defining both bone erosion and brain tissue abnormality.

To explore the causes and mechanisms of ureteric stones complicated by ureteric polyps and to improve the diagnostic and therapeutic effects. Methods:Clinical data of 58 cases of patients with ureteric stones complicated by ureteric polyps admitted from 1983 to 1999 were analyzed retrospectively. Results :41 cases underwent ureterolithotomy, as well as resection of stricture ureter and ureterourteroanastomosis. 17 cases re ceived ureterolithotomy and electronic resection or electronic ablation of polyps. 48 cases had indwelling ureteral stents. Symptoms of 58 cases disappeared completely after surgical operations. Conclusions:In cases that the stone has remained in the ureter for a long time, or in cases that the degree of hydronephrosis did not meeting the size of the stones, the possibility of ureteric polyps should be taken into account

OBJECTIVETo determine the three-dimensional stability of C1-C2 after anterior approach screw fixation through C2 vertebral body to C1 lateral mass for C1-C2 instability or dislocation.

METHODSThe three-dimensional range of motion of atlantoaxial joint were measured in 16 human cadaveric specimens under four conditions: the intact state (1st group), odontoid fracture of type II (2nd group), instrumentation with posterior C1-C2 trans-articular screw fixation (Magerl technique) (3rd group) and anterior approach screw fixation through C2 vertebral body to C1 lateral mass (4th group) respectively.

RESULTSThere was generally significant difference between 1st group and other groups and between 2nd group and other groups by statistics analysis (P < 0.001). Range of motion significantly decreased in 3rd group and 4th group in all directions. There was generally no significant difference between the two methods by statistical analysis (P > 0.05).

CONCLUSIONAnterior approach screw fixation through C2 vertebral body to C lateral mass provides satisfied stability. It provides the equivalent effect to Magerl technique. It is a kind of reliable surgery choice for the treatment of instability or dislocation of C1-C2 joint.

Adult ; Atlanto-Axial Joint ; surgery ; Biomechanical Phenomena ; Bone Screws ; Cadaver ; Cervical Vertebrae ; surgery ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Internal Fixators ; Male