Aim To prepare the dextran derivative nanoparticles with higher transfecting efficiency to mda-mb-435 human breast cancer cells.Methods The dextran 40 and the sodium periodate were reacted with different mole ratio to form the intermediate:oxidized dextrans having different aldehyde content.With different reactants matching,the spermine were grafted onto the oxidized dextrans mentioned above to form the dextran-spermine imine conjugates which were reduced by the excess reducing agent NaBH4 to produce the end products 9 species of dextran derivative nanoparticles(extran-spermine amine conjugates).The dextran derivative nanipartical of the highest gene transfecting efficiency was selected from the 9 species of dextran derivative nanoparticles by the experiment of gene transfection,using the pEGFP-C1 plasmid as the reporter,the mda-mb-435 human breast cancer cells as the transfected cells.Results The dextran derivative nanoparticles with the highest transfecting efficiency was the one that had the highest content of spermine grafted onto the oxidized dextran40 obtained by the reaction between sodium periodate and dextran40 with the mole ratio of 0.8.Conclusion The dextran derivative nanoparticles of the highest transfecting efficiency to the mda-mb-435 human breast cancer had been prepared.

ABSTRACT AIM To investigate the effect of ch-itosan on protein kinase C (PKC) activitives and diacyl glycerol(DAG)concentration in murine peritoneal macrophages. METHODS A new ion-pair reversed-phase high liquid chromatography and ra-dioimmunoassay were used to determinate the activity of PKC and the concentration of DAG respectively. RESULTS ( 1) Chitosan induced activation and translocation of PKC in murine peritoneal macrophages. The peak time was 25 min and the activity of PKC came back the basic level at 1. 5 h. (2)Chitosan increased the production of DAG in murine peritoneal macrophages. The peak time was 30 s and the concentration of DAG came back the basic level at 3 min. CONCLUSION The im-munopoteniating effect of chitosan may be associated with the channel of DAG /PKC.

Objective To compare the clinical outcomes of cervical vertebral reconstruction using titanium mesh and nano-hydroxyapatite and polyamide 66 (n-HA/PA66) mesh as two different kinds of artificial vertebrae. Methods From January 2008 to June 2009 had surgically treated 37 cases of cervical spondylosis with anterior corpectomy for decompression, and artificial vertebrae implanted combined platescrews fixation for cervical reconstruction and fusion. As a vertebral substitution, titanium mesh were implanted in 25 cases ( group A ), and the others (12 cases, group B) were implanted with n-HA/PA66 mesh. The height of fusion space immediately postoperatively and the implanted condition were observed. The fusion rate and the JOA score were recorded to compare the clinical outcomes. Results Two groups received 6-20 ( 15.4 ±4.2 )months follow-up. When follow-up to the end grafts were already fusion. The preoperative JOA score in group A,B [(8.40 ±0.96) scores vs. (8.33 ±1.07) scores(P ＞ 0.05)] were increased to (14.36 ±0.86)scores and (14.83 ±0.71) scores after postoperatively (P＞ 0.05),but there were statistically significant differences between preoperative and postoperative each group (P＜0.05). There were 6 cases artificial vertebrae in group A and 1 case in group B subsided asymptomaticly with 100% choiceness rate. Conclusion These two kinds of artificial vertebrae could show similar outcomes in cervical vertebral reconstruction, but the n-HA/PA66 mesh has a lower subsidence rate.

Objective To discuss the effects of Modic II changes on clinical outcomes of discectomy for lumbar disc herniation (LDH) with low back pain associated with unilateral sciatica. Methods Sixty-five cases of LDH with low back pain associated with unilateral sciatica received single segment discectomy during January 2007 to January 2009.There were 30 cases with Modic Ⅱ changes in group A, 35 cases without Modic Ⅱ changes in group B. Assessed the clinical outcomes by using MacNab analyzing system and visual analog scale (VAS). Results Two groups of the postoperative clinical symptoms had significant relief, the follow-up of 12 - 36 months, average (20.6 ± 7.5) months. MacNab efficacy evaluation by group A of optimal 10 cases (33.33%), good 17 cases (56.67%), general 3 cases (10.00%). Group B optimal 28 cases (80.00%), good 5 cases (14.29%), general 2 cases (5.71%), there was significant difference in fine rate (P<0.05). Preoperative group A VAS was ( 8.67 ± 0.30) scores, group B was (8.60± 0.32 ) scores (P>0.05). Postoperative group A VAS was (2.63 ± 1.30) scores,group B was (1.09 ±0.50) scores (P< 0.05). Conclusion Modic Ⅱ changes may be one of the reasons which cause the residual low back pain after the discectomy for LDH.

AIM: To evaluate the effects of NS-398, a cyclooxygenase-2(COX-2) inhibitor, on the proliferation and apoptosis in HepG2 cells. METHODS: The effects of NS-398 on the proliferation of HepG2 cells was evaluated by MTT. DNA fragmentation gel analysis was used to analyze the apoptotic cells; DNA ploidy and apoptotic cell percentage were examined by flow cytometry. Furthermore, the expression of COX-2 and Bcl-2 mRNA was identified by competitive RT-PCR. RESULTS: NS-398 inhibited cell proliferation and induced apoptosis in HepG2 in a concentration-dependent manner. DNA ploidy analysis showed that S phase cells were significantly decreased with NS-398 concentration increasing. The quiescent G_0/G_1 phase was accumulated with decreasing of Bcl-2 mRNA. Whereas NS-398 had no effect on the expression of COX-2 mRNA, no correlations were found between COX-2 mRNA and the HepG2 cell proliferation and apoptosis induced by NS-398 (r=0.056 and r=0.119, respectively). CONCLUSION: NS-398 significantly inhibits the proliferation and induces apoptosis in HepG2. Mechanisms may be involved in accumulation of quiescent G_0/G_1 phase and decrease in Bcl-2 mRNA expression, but independent to COX-2 mRNA expression. [

Objective: To study the changes of andrographolides in the drying process of the extract of Andrographis paniculata Nees. Methods: HPLC method was applied to analyze andrographolide and 14 deoxy 11, 12 didehydroandrographolide in the process. Results: The content of andrographolide descended rapidly in the whole drying process, while the content of 14 deoxy 11, 12 didehydroandrographolide ascended at first 12 hours, declined in content was slowly to follow. Conclusion: Baking temperature is not the only main factor to stimulate the transformation of andrographolide and 14 deoxy 11, 12 didehydroandrographolide.

OBJECTIVE:To optimize the formulation of azithromycin sustained-release vaginal suppository(ASVS).METHODS:The ASVS was prepared using S-40 as suppository base,HPMC as sustained-release material and glycerine as humectant.The formulation of ASVS was optimized by orthogonal design with accumulated release percentage and hardness as indexes and the amount of 8% HPMC and glycerol as factors;meanwhile,a verification test and the fitting of in vitro drug release model were performed.RESULTS:The optimized formulation ASVS was as follows:azithromycin 6 g,8% HPMC 23.52 g,glycerol 29.40 g,ethanol absolute 3 g,ethylparaben 0.59 g,and S-40 294 g.Three batches of suppositories prepared under the optimized formulation reached a mean content of 99.5%,with hardness up to the standard,showing good reproducibility and homogenicity in drug release in vitro.The accumulative release rates of all samples were greater than 98% at 180 min,and the release dynamics in vitro were in line with Higuchi equations.CONCLUSION:The optimized ASVS is feasible in formulation,stable and reproducible in preparation technology and up to the standard for sustained-release preparation.

ObjectiveTo investigate the distribution of high-intensity zone (HIZ) of lumbar intervertebral disc in patients with low back and/or leg pain,and analyze its related factors.MethodsSix hundred and twenty-eight patients with low back and/or leg pain were examined by MRI scan from June 2009 to August 2010.According to the diagnostic criteria of HIZ,the features of distribution of HIZ on age,segment and degree of intervertebral disc degeneration were analyzed retrospectively.ResultsAmong 3140 intervertebral discs of the 628 patients,172 cases (27.39％,172/628) and 206 discs (6.56％,206/3140)were involved with HIZ.There was no significant difference between men and women [26.38％(86/326)vs.28.48％(86/302)] (P=0.556).HIZ occurred more often [40.22％(72/179)] in those patients between 40 and 49 years of age.The incidence of HIZ at the segments from L1-2,L2-3,L3-4,L4-5,L5-S1 was 0.80％(5/628),2.07％(13/628),2.07％(13/628),14.01％(88/628) and 13.85％(87/628) respectively.In cases with and without HIZ,the incidence of intervertebral disc degeneration up to grade V was 49.03％(101/206) and 23.76％(697/2934) respectively (P ＜ 0.01 ).HIZ was correlated with age,degree of intervertebral disc degeneration and disc segment (r =-0.040,P=0.025 ;r =0.217,P＜ 0.01 ;r =0.179,P＜ 0.01 ).Conclusions HIZ is correlated with age,degree of intervertebral disc degeneration and disc segment.Intervertebral disc degeneration plays the most important role in the occurence of HIZ.HIZ_mainly occur in L4-5 and L5-S1 segment and in those between 40 and 49 years of age.

Objective To evaluate the effect of various reduction degrees of large posterior malleolus fracture on contact condition of the tibiotalar joint using biomechanical and finite element methods.Methods Six fresh adult cadaveric calf-foot were tested with different loadings in neutral position,10 degree of dorsiflexion,and 15 degree of plantar flexion.Models included the intact ankle and posterior malleolus fracture (not fixed,fixed with 1 mm step-off,fixed with 2 mm step-off,fixed with 1 mm gap,or fixed with 2 mm gap).Tibiotalar joint contact area,contact pressure,and peak pressure were measured using the Tek-Scan pressure sensor.In addition,a three-dimensional finite element model of the ankle joint was established and tested under the simulated biomechanical conditions.Results Mter a 500 N axial loading in the intact ankle,contact area of the tibiotalar joint was (4.94 ± 0.67) cm2 and loading strength of the tibiotalar joint was (83.4 ± 2.7) ％ of total loading strength.Contact area of the tibiotalar joint in the fracture model fixed with 1 mm gap demonstrated no significant difference in all loading conditions compared with the intact model.In 15 degree of plantar flexion,contact area of the tibiotalar joint differed between the fracture model fixed with 1 mm step-off and the intact model (P ＜ 0.05).In multiple loading conditions,contact area of the tibiotalar joint differed between the fracture models fixed with 2 mm step-off or 2 mm gap and the intact model (P ＜ 0.05).Results of finite element analysis were conformed to the related biomechanical study.Conclusions Anatomic reduction should be pursued for large posterior malleolus fracture.If difficulty exists,fracture displacement should be within 1 mm step-off and 2 mm gap so as to restore the inter-joint congruity.

Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor(GR)binding domain.Methods The fragments of GR binding domain was amplified by RT-PCR,and then was cloned into pMD18-T.After being verified by sequencing,it was subcloned into the bait expression vector pGBKT7.Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot.Toxicity and self-activation of the bait protein were detected.Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully.The bait vector was transformed into AH109 yeast cells successfully,without toxicity or self-activation.The expression of the bait protein was confirmed by Western blot.Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.