The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.

Objective To detect genetic alterations in pleomorphic xanthoastrocytoma (PXA), and to investigate the mechanism of development of this neoplasm. Methods Three patients with PXA were studied. Comparative genomic hybridization (CGH) was performed to study chromosomal imbalances in PXA. Using immunohistochemical analysis, the expression of EGFR was detected in PXA. Results Using CGH analysis, genetic imbalance was detected on at least one chromosome for each case. One patient revealed multiple genetic alterations, including gains of 2p14-pter, 4p15-pter, 7p21-qter, 11q24-qter, 12 and 15q14-qter,as well as losses of 8p11.2-pter, 9p11-p23, 10p12-pter, and 13q14-qter. This patient experienced tumor recurrence and died one year later. Gain on Chromosome 7 and loss on Chromosome 8p were demonstrated in 2 of the 3 patients. Immunohistochemically, no EGFR positive reaction was found in all cases. Conclusion Detection of genetic alterations is very important in understanding the pathogenesis of PXA.

Objective To investigate the clinicopathologic features,diagnosis,differential diagnosis and treatment of sclerosing angiomatoid nodular transformation(SANT). Methods The clinical data,pathologic characteristics,immunophenotype and postoperative follow-up of SANT were analysed. Results There were no specific findings in the clinical manifestations of the 4 cases of SANT.Grossly,the cut surface of the masses was gray-white and vague nodularity was observed.Microscopically,it was characterized by the multinodular angiomatoid appearance in a fibrosclerotic stroma.The nodules were composed of slit-like,sinusoid-like vascular spaces and were interspersed with a population of spindly or ovoid cells.It was revealed by immunohistochemistry that the expression of CD34 in some vessels' endothelial cells was positive,and CD8 was negative.While in another vessels' endothelial cells,CD8 was positive and CD34 was negative.The expression of SMA,Actin,Vimentin,Collage IV and CD68 was positive in all of the 4 cases,while that of CD21,Desmin and NSE was negative.No relapse or metastasis was found during the follow-up.Conclusion SANT is a rarely encountered benign lesion of the spleen,which should be distinguished from the malignant tumor of the spleen.The diagnosis counts on the pathologic and immunohistochemical findings.It could be cured by splenectomy with a favourable prognosis.

AIM: To compare and analyze the stereopsis of intermittent exotropia children under the different backgrounds of dynamic stimuli and static stimuli.
METHODS: We collected 56 children ( male 26, female 30 with intermittent exotropia at the age from 5y to 12y and examined their stereopsis under the different backgrounds of dynamic stimuli and static stimuli using a multidimensional sense perception training software. The differences between the dynamic stereopsis and static stereopsis were compared.
RESULTS: Totally 17 cases ( 30%) had both dynamicstereopsis and static stereopsis, 39 cases ( 70%) had either dynamic or static stereopsis deficit, only 10 cases ( 26%) had dynamic stereopsis, 25 cases ( 64%) static stereopsis left and 4 cases ( 10%) were without any form of stereopsis. The positive rate of dynamic stereopsis was better than the positive rate of static stereopsis, with statistical significance (P<0. 05).
CONCLUSION: Dynamicstereopsis is better than the static stereopsis to intermittent exotropia children.

Pulmonary arterial hypertension (PAH), also named as a cancer of cardiovascular disease, is a rare disease and has complicated pathogenesis. Recently, there are more understandings of PAH pathogeneses. According to the pathogenesis and active pathways, the clinically used drugs are classified into several groups incluidng prostacyclin analogues and prostacyclin receptor agonists, endothelin receptor antagonists, phosphodiesterase-5 inhibitors, soluble guanylate cyclase inhibitors, etc. To elevate the efficacy of the drugs, numerous drug delivery systems are developed. This review mainly summarized the pathological mechanism of PAH, drugs and drug delivery approaches in the treatment of PAH.

AIM: To analyze the effect of pigment epithelium derived factor (PEDF) on nitrogen monoxide (NO) and expression of cysteine-containing, aspartate-specific proteases-3 (caspase-3) in retinal tissues of model rats with optic nerve crush injury.METHODS: A total of 60 SD rats were randomly divided into the blank control group, model group and PEDF group, with 20 rats in each group.Except the blank control group, the optic nerve crush injury rat models were established in the other groups, and left eyeballs were taken as samples.After successfully modeling, the model group were treated with intravitreal injection of 5μL of balanced salt solution while PEDF group were treated with intravitreal injection of 5μL of PEDF (0.2μg/μL).Two weeks later, the retinal tissues were collected, and changes of shape were observed under microscope after HE staining.The changes of NO level were measured by colorimetry assay, the expression of caspase-3 mRNA and caspase-3 protein was detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot.RESULTS: HE staining showed that retinal tissues of the blank control group arranged neatly and clearly.Retinal ganglion cells (RGCs) arranged in a monolayer, and cells were oval, uniform in size and distribution, the cell nuclei were clear, closely arranged, with clear boundaries.The retinal tissues of the model group were sparse in shape, RGCs showed vacuolar changes, the overall number of cells was reduced, and cell nuclei of residual RGCs showed pyknosis and uneven staining.RGCs in PEDF group were with slightly edema and arranged closely, and the degree of injury was significantly milder than that in the model group.Levels of Caspase-3 mRNA and protein and NO levels in the three groups showed the model group > PEDF group > blank control group (all P < 0.05).CONCLUSION: The application of PEDF can down regulate the expression of Caspase-3 and NO in rates with optic nerve injury and reduce RGCs injury.

Thorough excavation and artful utilization of various kinds concrete and invisible learning resources contribute to the cultivation of excellent postgraduates.In postgraduate education of anesthesiology Xuzhou Medical College utilizes time,network,technique platform,research outcome,self-potentiality and clinical patients resources,which produces an active effect and has important instructional significance.

Objective To study the characteristics of event-related potentials P300 in patients with anxiety disorder(AD). Methods P300 tests were carried out in 30 patients with AD and 30 healthy adult controls. ResultsPatients with AD had significantly delayed P3 latency ([326?16] ms vs [339?19]ms, P

Objective To investigate the regulatory effects of tuberculin on growth and apoptosis of liver cancer and lung cancer cell lines. Methods HePG2(liver cancer) and A549(lung cancer) cell lines were treated with TB supernatant(TB-SN) with different concentrations. Cell viability was detected by using LIVE/DEAD Viability/Cytotoxicity cell kits including specific fluorescence primer,and cell apoptosis was detected by Vybrant apoptosis assay. Results After treatment with 5% TB-SN for 5 d,cell apoptosis was significantly increased in HePG2 and A549 cell lines.Cell growth of HePG2 and A549 cell lines was inhibited after treatment with TB-SN. Conclusion Tuberculin can induce cell apoptosis and inhibit cell growth of liver cancer and lung cancer cell lines.

Objective To investigate the expression of macrophage migration inhibitory factor(MIF) in renal tissue of children with Henoch-Schonlein purpura nephritis(HSPN),and its correlation with clinical indexes and pathological changes,and to explore its effect on the pathogenesis of HSPN.Methods According to the clinical manifestation,60 children with HPSN were divided into only purpura group,mixed group and HSPN group.MIF concentration of Henoch-Schonlein purpura(HSP) groups and healthy control group were detected with enzyme linked immunosorbent assay(ELISA).MIF protein expression and the marker of human macrophage(CD68) in renal tissues of HSPN and normal control group were detected with immunohistochemistry method.The total urine protein for 24 hours and urinary N-acetyl-beta-D-glucosaminidase (NAG) level were detected with laboratory routine method.Results MIF concentration in mixed group and HSPN group were significantly higher than that in only purpura group and healthy control group(Pa