Objective: To establish an in vitro model of brain blood barrier (BBB) using cultured mouse brain microvascular endothelial cells (BMVEC). Methods: Mouse BMVEC were seeded on micro pore membrane of gelatin coated cell culture insert and cultured to confluence. The establishment of BBB was preliminary judged by a 4 h water leaking test. The tight junctions between BMVEC were demonstrated by scanning and transmission electron microscope. The transendothelial electrical resistance(TEER) over BMVEC was measured. The permeability of Horseradish peroxidase (HRP) through the BBB was analyzed and the effect of RMP 7 on permeability of the BBB was investigated. Results: The 4 h water leaking test became positive when BMVEC were cultured to confluence. By scanning and transmission electron microscope, the tight junctions were demonstrated on confluent BMVEC. The TEER over BMVEC monolayer increased 3.2 and 7.68 times and the permeability rates for HRP were 13.4% and 6.7% respectively, as compared with sub confluent BMVEC and human umbilical vein endothelial cell monolayer(HUVEC). The HRP permeability rate in the model of BBB increased 2.7 times after treatment with RMP 7. Conclusion: The established in vitro model of BBB has basic characteristics of BBB in vivo , and is suitable for central nervous system (CNS) drug research over BBB.

Objective: To study the 3D positional changes occurring to anchor molars of 37 patients treated with MBT appliance. Methods: Thirty seven patients who needed maximum upper molar anchorage were divided into two treatment groups at random, group Ⅰ was comprised of 19 patients whose anterior teeth were retracted en masse, and group Ⅱ included 18 patients whose anterior teeth were retracted by two step. All the patients were treated with MBT appliance and headgear. Dental casts recorded before and after treatment were analyzed by YM 2115 three dimensional digitizer and related software. The changes of anchor molar tip,torque and rotation were compared between the two groups. Results: During the treatment, average anterior movement of the upper first molars was 4.53 mm, average extrusion of the upper first molars was 1.53 mm; the crown of the upper first molar tipped forward (6?4)?,torque buccally (3?6)?,and mesiolingually rotated 3.15?; In en mass group, the crown of the upper first molar tipped more mesially than the two step group, and the difference was statistically significant. Conclusion:Almost all the upper first molars showed the tendency to rotate and tip mesially and torque bucally; there was no statistically significant difference between the two treatment groups when upper molar positional changes were concerned except for the mesial tip of the crown.

Objective: To study the chemical constituents of the stem bark of Albizia julibrissin Durazz. Methods: Chemical constituents were isolated by the repeated chromatography methods and their structures were identified by spectral analysis. Results: Five compounds were obtained as follows: 3 O [? D xylopyranosyl (1→2) ? D fucopyranosyl (1→6) ? D glucopyranosyl] 21 O (6S) 2 trans 2 hydroxy methyl 6 methyl 6 O ? D quinovopyranosyl 2,7 octadienoyl acacic acid 28 O ? D glucopyran osyl (1→3) [? L arabinofuranosyl (1→4)] ? L rhamnopyranosyl (1→2) ] ? D glucopyranosyl ester (1). acacigenin B (2), julibrotriterpenoidal lactone A (3), machaerinic acid lactone (4), acacic acid methylester (5). Conclusion: Compound 1 was isolated directly from this plant for the first time.

0.1%EDTA≈5% HP ? CD≈1%lecithin. Conclusion: The three methods have good correlation.

Objective: To investigate the expression of PDCD5 in tissues of normal cervix, CIN Ⅰ-Ⅲ,cervical cancer and explore the relationship between PDCD5 and cervical cancer.Methods: After we defined the most fitful condition, tissues from 18 cases of normal cervix, 19 of CIN Ⅰ, 18 of CIN Ⅱ, 20 of CIN Ⅲ and 18 of cervical cancer were defined by indirect immunohistochemical technique. Positive expression rates and intensity of PDCD5 protein were investigated by observing under microscope and analyzing with computer imaging technique. The results were analyzed with one way anova. Results: The results of immunohistochemical staining showed that the percentage of strong positive cells in normal cervical tissue and CIN Ⅰ were significant higher than those of CIN Ⅱ, CIN Ⅲ and cervical cancer. On the whole of the condition of immunohistochemical staining, the expressions of PDCD5 were downregulated along the progression of cervical atypical epithelia, but that in CIN Ⅰ was upregulated.The ODs of normal cervix,CIN Ⅰ-Ⅲ,cervical cancer were 0.322,0.366,0.287,0.252,and 0.206 respectively.The intensity of each group showed obvious differences.Conclusion: We found that the expression of PDCD5 was upregulated in CIN Ⅰ and downregulated in CIN Ⅱ, CIN Ⅲ and cervical cancer. It suggests that PDCD5 is an important apoptosis regulating factor in the occurrence of cervical cancer.

Objective:To study the role of interleukin 9 in the airway inflammation from patients with COPD. Methods: Induced sputum was obtained from 30 COPD patients with stable disease(group A) ,31 asthmatics patients with stable disease(group B) and 15 healthy individuals(group C). IL 9,IL 5 and IL 8 in sputum supernatants were measured by sandwich enzyme linked immunosorbent assay(ELISA) and IL 9 positive expression and quantitative analysis were conducted by Streptavidin peroxidase method and image analysis technology. Results:The levels of IL 9 in group A and B were all significantly higher than those in groups C. IL 9 positive expression mainly located in the cytoplasm of macrophages. The positive rates of IL 9 in group A and B were all significantly higher than that of group C(? 2=20.821, 19.908, P

0.20), but decreased the number of nNOS positive cells in central canal (79% of H saline, P

Objective: To obtain the antibody against N terminal of 1A6/DRIM, and thereafter get the profile of 1A6/DRIM expression in different cell lines. Methods: The N terminal of 1A6/DRIM (aa 577 714) was cloned into pGEX 4T 3. Multiple antigenic peptides (MAPs)(aa638 661) was synthesized as the antigen with Fmoc/PyBOP method. Rabbits were immunized by injecting the MAPs and the immunized sera were analyzed with ELISA and Western Blot. The Western Blot and immunofluorescence were performed to analyze the expressing profiles of the 1A6/DRIM in different tumor cell lines. Results: The antibody specifically recognized the full length of 1A6/DRIM as a 310 kDa band, which was also recognized by C terminal monoclonal antibody shown by Western Blot. 1A6/DRIM is expressed in multiple tumor cell lines and mainly located in the nuclei. Conclusion: Preparation of the antibody with MAPs is a useful technique when the fusion proteins can not be induced in E coli . The antibody we got via MAPs has supplied a good tool for further studies on the functions of the novel gene 1A6/DRIM.

Objective: To analyze the expression of mecp2 gene at mRNA and protein;Cerebral cortex level in the cerebral cortex of the normal Wistar rat throughout development. Methods: We chose the 15th day (E15), 17th day (E17), 19th day (E19) of embryo period, the day of birth (P0), the 7th day (P7), the 14th day (P14), the 28th day (P28) of postnatal period, and adulthood as analyzing time points. The expression of mecp2 gene at mRNA level was analyzed by real time PCR and Northern blot. The expression of MeCP2 protein was analyzed by Western blot. Results: There was one type of mecp2 mRNA transcript (approximately 10 kb) expressed in the cerebral cortex of the normal Wistar rat. The expression level of mecp2 mRNA varied subtly during the development. There was one type of MeCP2 protein (75 000) expressed in the cerebral cortex of the normal Wistar rat. The expression level of MeCP2 protein remained the lowest on E15, from E19 to adulthood the expression levels of MeCP2 protein increased dramatically compared with that on E15. From P7 to adulthood, the differences of expression between two time points were subtle. Conclusion: The expression level of MeCP2 protein increases as the neurons in the cerebral cortex of normal Wistar rat grow mature. This indicates that MeCP2 protein is very important to neuron's maturation, and probably has relationship with maintaining maturation state of neurons.

Objective: To study the protection of IL 1ra in cultured developing neuron injury following Mg 2+ free induced epileptiform discharges. Methods: Rat embryo cortical neurons cultured for 6 d and 17 d were directly exposed to Mg 2+ free media, or pretreated with IL 1 receptor antagonist or NMDA receptor antagonists before being exposed to Mg 2+ free media, and then returned to regular media.MTT assay was used to study mitochondrial function injury, laser scanning confocal microscope to measure [Ca 2+ ]i, and real time RT PCR to detect gene mRNA expression. Results:(1) MTT conversion rates were higher in neurons pre and co treated with 10 mg/L IL 1ra than those of neurons with only Mg 2+ free treatment in neurons cultured for 17 d, but not in neurons cultured for 6 d.(2) [Ca 2+ ]i was lower in neurons pre and co treated with 10 mg/L IL 1ra than those of neurons with only Mg 2+ free treatment, either in neurons cultured for 6 d or in neurons cultured for 17 d, and the effects of IL 1ra on [Ca 2+ ]i change were different between neurons cultured for 6 d and neurons cultured for 17 d.(3) Pre and co treated with 10 mg/L IL 1ra NR1 mRNA expression increase induced by Mg 2+ free treatment was decreased, either in neurons cultured for 6 d or neurons cultured for 17 d, and this effect showed no difference between neurons cultured for 6 d and 17 d; Pre and co treated with 10 mg/L IL 1ra NR2A mRNA expression increase induced by Mg 2+ free treatment in neurons cultured for 17 d was decreased, and NR2A mRNA expression showed no difference between IL 1ra group and age matched control group, but have no effect on neurons cultured for 6 d; Pre and co treated with 10 mg/L IL 1ra have NR2B mRNA expression increase induced by Mg 2+ free treatment was not affected, either in neurons cultured for 6 d or neurons cultured for 17 d. Conclusion:Neuroprotection of IL 1Ra in seizure induced injury is age dependent. The mech anism of the neuroprotection of IL 1Ra includes down regulation of [Ca 2+ ]i and others.