Objective To explore the characteristics of WAS gene mutation in Wiskott-Aldrich syndrome (WAS). Methods The clinical data of one infant with WAS were retrospectively analyzed. All exons and flanking sequences in WAS gene were detected by PCR. Results A 5-month-old boy, who has a history of eczema and recurrent infection, was hospitalized for thrombocytopenia. CD8+ and CD4+T cell were increased while CD19+B cell was normal. Bone marrow cytology suggested megakaryocyte mature hindrance. WAS gene detection found C.880 A?>?G (p.Ile294Val) mutation, but no mutations were found in parents. This site was a pathogenic mutation predicted by Polyphen 2 software and SIFT software. Besides, sequence conservation analysis of different species found it was a conservative site and structural prediction analysis revealed it may affect the normal protein structure. This site of mutation has not been reported before. Conclusions Gene detection can make early diagnosis of WAS and C.880 A?>?G (p.Ile294Val) is a new mutation.

Objective:To explore the depressive symptom of nurses and its relationship with trait coping style,role perceive and personal control.Methods:425 nurses in the 2nd Hangzhou general hospital completed a battery of questionnaires consisted of part items of SCL-90,Trait Coping Style Questionnaire(TCSQ) and Role Perceive and Personal Control Scale.Results:There are significant correlation among the SCL-90 score,the coping styles,and the control feeling and role conflict.Based on depression factor ,factors of negative coping style,control feeling,role conflict and positive coping style were included in stepwise regression equation.Conclusion:Depressive symptom of nurses was related to trait coping style,role perceive and personal control.

Objective:To investigate the relationship among college student’s depression symptoms, problem solving appraisal and perceived stress.Methods:275 college students completed questionnaires, including CES-D, Chinese Perceived Stress Scale (CPSS) and Chinese Problem Solving Inventory (CPSI). Results: Among these students, 52% reported on the level of health risk stress (HRS), 55.3% had depression symptoms. There were significant correlation among depression symptoms, perceived stress and problem solving appraisal. The causal relationship between them were identified with the path analysis method, problem solving in special, seems to have both direct and indirect effect on the prediction of depression and on perceived stress. Conclusion:There is a high level of college student perceived stress and depression symptoms. Problem solving appraisal is an important factor for both of them.

AIM: The synergistic effect of basic fibroblast growth factor (bFGF) and endothelin- 1 (ET-1 ) on rat aortic vascular smooth muscle cells(VSMC) proliferation was observed. The possible mechanism of the synergism was also investigated. METHODS: BrdU incorporation and cell counting method were adopted to value the pro - proliferative effect of VSMC. Western- blotting was used to observe the variation of bFGF and FGFR - 1 isoforms expression. RESULTS: bFGF and ET- 1 could promote VSMC proliferation separately, and synergistically in combination. The synergism was dose - and time - dependent. ET- 1 increased all the three bFGF isoforms and FGFR - 1 protein level in dose - and time- dependent manner. In addition, after exhaustion of intracellular PKC, the upregulation effects of ET- 1 on bFGF and FGFR - 1 expressions in VSMC both inclined. CONCLUSION: bFGF and ET- 1 had synergistic effect on VSMC proliferation. ET- 1 may increase the responsiveness of VSMC to bFGF through modulation of bFGF isoforms together with FGFR - 1, which was PKC - dependent.

Objective To investigate the developmental profiles on surface expression and distribution of NMDA receptor clusters especially on dendritic structures in cultured hippocampal neurons of rats. Methods A vector expressing green fluorescent protein N-terminally tagged NR1a subunit(GFP-NR1a) was generated and transfected into cultured hippocamplal neurons at 5 days in vitro (DIV 5).Surface expressed GFP-NR1a containing NMDA receptor clusters was then labeled in living neurons by using anti-GFP primary antibody followed by Cy3-conjugated secondary antibody.Furthermore,distribution of the surface clusters was observed on the detailed dendritic structures visualized with co-expression of cyan fluorescent protein(CFP). Results Punctate green flourescence clusters can be seen throughout the neurons transfected with GFP-NR1a and surface staining of living neurons showed that the most of these flourescence clusters on dendritic arbors were surface expressed in mature neurons.In parallel,no significant difference was found in terms of density of the surface NMDA clusters on dendritic trees in neurons at different developmental stage.Interestingly,at DIV 7,surface NMDA receptor clusters were mostly observed on the dendritic shafts,but rarely on filopodia.In contrast,two weeks later about 50% of clusters were found located at the dendritic spines.Conclusion This study shows that surface NMDA receptor clusters are distributed in a differentiated way related to dendritic structures during neural development.Especially at very early developmental stage,no clusters was found presented on filopodia,instead they were extensively clustered with a density estimated to be not significantly different from mature neurons.Therefore,our results suggested that NMDA receptors are likely to be incorporated or recruited to postsynaptic sites in the form of pre-expressed surface cluster on dendritic shafts during glutamatergic synapse formation.

Objective To review the physiological characters and potential function of chondroitin sulphate proteoglycan,CSPG(NG2) glial cells in the central nervous system. Methods In conclusion of the NG2 glial properties and the global, novel researches on NG2 cells. Results Cells that express the NG2 proteoglycan comprise a unique population of glial cells in the central nervous system.Study on their role in the CNS becomes more and more popular.Previous researches mainly focus on the differentiation and distinction.Now the relationship between NG2 glial cells and neurons,and the synaptic plasticity is becoming a hot topic.Conclusion This review referred the research development of NG2 glial cells,including the differentiation,migration,their interaction with neurons,and the potential role in glial cell regeneration in disease systematically.

Objective To investigate the therapeutic effect of Chinese herbal medicine for seminal delayed liquefaction(SDL).Methods Two hundred and ninety SDL patients were randomized into two groups.Group A(N=230)received oral use of yin-nourishing and blood-activating herbal medicine(mainly composed of Radix Ophiopogonis,Radix Scrophulariae,Fructus Ligustri Lucidi,Herba Ecliptae,Radix Salviae Miltiorrhizae,Flos Carthami,Rhizoma Sparganii,Eupolyphaga seu Steleophaga),and group B(N=60)received intramuscular injection of chymotrypsin and oral use of vitamin C and zinc gluconate.The therapeutic effect in the two groups were evaluated after treatment,and the changes of main parameters of seminal liquid as well as the seminal plasma contents of prostate specific antigen(PSA),acid phosphatase(AP),citric acid(CA)and zinc were observed before and after treatment.Results In group A,156 patients were cured,57 effective,17 ineffective and the total effective rate was 92.6%;in group B,21 patients were cured,17 effective,22 ineffective and the total effective rate was 63.3%.The Therapeutic effect was better in group A than that in group B(P

AIM: To get large amounts of pure antigens to raise specific antibodies and to perform quantifications.METHODS: CYP2B6 (cytochrome P) cDNA fragments was ligated into BamHI restricted PGEX-3b to generate recombinants PGEX/2B6. We identified recombinants PGEX/2B6 by EcoRI digestion. The expression of fusion proteins were induced by adding isopropyl-thiogalactoside(IPTG). Several clones showed high-level expression of fusion proteins. Insoluble proteins was isolated from the bacteria and the fusion proteins was recovered and purified from a preparative (2mm) SDS-PAGE. The polyarcrylamide gel containing the fusion proteins glutathione S-transferase(GST-2B6) were used to immunize BALB/C mice from which polyclonal ascites fluid was prepared. The purified fusion proteins GST-1A1(GST fusion protein of CYP1A1 cDNA246～386aa expressed in this library ,purified by preparative SDS-PAGE), GST-2B6 were used to test the specificity of 2B6pAb. RESULTS:Fusion proteins constructed between GST and CYP2B6 was expressed in Escherichia coli DH5?. Mouse antibodies are raised against the fusion proteins GST-2B6. 2B6pAb was fond to be specific antibody.CONCLUSION:Recombinant PGEX/2B6 were constructed and purified fusion proteins GST-2B6, and specific 2B6pAb were obtained.

AIM: The synergistic effect of basic fibroblast growth factor (bFGF) and endothelin-1 (ET-1) on rat aortic vascular smooth muscle cells(VSMC) proliferation was observed. The possible mechanism of the synergism was also investigated. METHODS: BrdU incorporation and cell counting method were adopted to value the pro-proliferative effect of VSMC. Western-blotting was used to observe the variation of bFGF and FGFR-1 isoforms expression. RESULTS: bFGF and ET-1 could promote VSMC proliferation separately, and synergistically in combination. The synergism was dose- and time-dependent. ET-1 increased all the three bFGF isoforms and FGFR-1 protein level in dose- and time-dependent manner. In addition, after exhaustion of intracellular PKC, the upregulation effects of ET-1 on bFGF and FGFR-1 expressions in VSMC both inclined. CONCLUSION: bFGF and ET-1 had synergistic effect on VSMC proliferation. ET-1 may increase the responsiveness of VSMC to bFGF through modulation of bFGF isoforms together with FGFR-1, which was PKC-dependent.

Objective To investigate whether the partially C-terminal deletion of NR2A subunit alters the surface expression and channel function of NMDA receptors in both HEK293 cells and cultured hippocampal neurons of rats. Methods Four plasmids for NR2A mutants with N-terminally GFP-tagged and C-terminal deletion NR2A?C1(?897L-1017S),NR2A?C2(?1024D-1142P),NR2A?C3(?1149D-1347G),and NR2A?C4(?1354S-1464V) were generated,and transfected into HEK293 cells and hippocampal neurons in culture.Surface staining was performed using anti-GFP antibody and Cy3 conjugated secondary antibody.Glutamate evoked currents were also detected using whole-cell recording. Results Positive surface staining was found for all the HEK293 cell co-transfected NR1-1a/NR2A?C1,NR1-1a/NR2A?C2,NR1-1a/NR2A?C3 or NR1-1a/NR2A?C4,and quantitative analysis showed no significant decrease in surface expression level when compared to that from NR1-1a/NR2A transfection.Glutamate-evoked currents were recorded in HEK293 cells co-transfected with NR1-1a/NR2A?C2 or NR1-1a/NR2A?C4.Surface expression level of NMDA receptor clusters on dendrites was significantly decreased in the neurons transfected with NR2A?C1,NR2A?C2 or NR2A?C3 than in those transfected with NR2A.Conclusion C-terminal deletion of NR2A subunit differentially effects surface expression of NMDA receptors in HEK293 cells and in hippocampal neurons in culture.