s Objective To observe the changes in the expression of ?-amyloid precursor protein (?-APP) in selected brain areas in early stage of diffuse axonal injury (DAI) in rats. Methods DAI was induced in SD rats by a self-made rotating injury device. 16 rats in injury group were sacrificed by decapitation at 3 hours and 6 hours post-trauma, while the other 8 rats in sham injury group were put to death after 24 hours. The samples of the rat brain were stained with ?-APP for immunohistochemistry in order to detect the early stage pathological changes in subcortex white matter, hippocampus, corpus callosum and brain stem at different time points. Results The ?-APP expression was positive at 3 hours and was particularly strong at 6 hours after injury. The results of semi-quantitative analysis showed a significant differentiation of 3 hours and 6 hours in the immuno-positive expression of brain stem, corpus callosum and hippocampus. Histological results demonstrated that DAI occurred in all rats of injury group. Conclusion ?-APP is a kind of sensitive and useful immunochemical marker for early DAI diagnosis, and it indicates axonal lesion or breakage.

Objective To determine whether cyclosporin A(CsA) could exert neuroprotective effects after diffuse axonal injury (DAI) in the rat. Methods Twenty four SD rats were randomly assigned into three groups: non injur group ( n =8); control brain injury group ( n =8), in which normal saline was given; and experimental group ( n =8), the injured rats were treated with CsA. The beam balance test device and Morris water maze were used to test for balance and cognitive performance. Results Control brain injury animals displayed severe defects in balance and cognitive performande after diffuse axonal injury. Compared with control brain injury animals, rats treated with CsA displayed better motor performance in beam balance tests and improved learning ability in the Morris water maze. Conclusions It is demonstrated that CsA exhibits substantial neuroprotective activity in a rat model of DAI. These findings support that CsA is a useful therapeutic agent in the treatment of DAI.

s Objective To observe the changes in neurochemical metabolites at early stage of DAI in rats with MRS technique. Methods DAI was induced by a self-made rotating injury device in Sprague-Dawley (SD) rats (n=16). MRS was used to evaluate the changes in neurochemical metabolites before-trauma, 2 hours and 3 hours after the trauma. The data were analyzed by statistical analysis system. Results MRS demonstrated that contents of both NAA/Cr and Cho/Cr of corpus callosum were decreased obviously 3 hours posttrauma, and differences between those before trauma and those after trauma were statistically significant. The content of NAA/Cr of the brain stem was declined at 2 hours also with significant difference compared with that before trauma. There was no significant difference in values between 2 and 3 hours. The content of Cho/Cr of the brain stem was significantly decreased 3 hours after the trauma. Conclusions MRS technique has a high sensitivity in diagnosing microscopic pathology following DAI and functional defect of neuron and axon, as shown by significant decrease in NAA/Cr and Cho/Cr at early stage (i.e. 2-3 hours) after trauma.

Objective To set up the reference standard of cyclovirobuxine D.Methods Thermal analysis,HPLC/MS,HPLC with terminal wavelength,HPLC with fluorescence derivation and with ultraviolet derivation,TLC and nonaqueous titration methods were applied to determine the content of cyclovirobuxine D control.Results Thermal analysis can not be used to analyse the purity of cyclovirobuxine D ,and HPLC/MS,HPLC with terminal wavelength,HPLC with fluorescence derivation and HPLC with ultraviolet derivation can obtain the same purity.Conclusion The methods used for the assay of cyclovirobuxine D control were practical.

Objective To investigate the role of 2B subunits-containing N-methyl-D-aspartate receptors (NR2B) in the arcuate nucleus in the development of inflammatory pain (IP) in rats.Methods One hundred and eight male Sprague-Dawley rats, aged 9 weeks, weighing 200-250 g, were randomly divided into 4 groups using a random number table: sham operation group (group S, n =47);group IP (n =47);dimethyl sulfoxide control group (group DMSO, n =7);selective NR2B antagonist Ro25-6981 group (group Ro25-6981, n=7).IP was induced by injecting complete Freund's adjuvant (CFA) 0.1 ml into the plantar surface of the left hindpaw.Ro25-6981 400 pmol was injected into the arcuate nucleus at 3 days after CFA injection.Seven rats in each group were selected for measurement of the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) at 1 day before CFA injection (T1) and at 2 days after CFA injection (T2), at 30 min before administration on 3rd day (T3) , at 30 min after administration on 3rd day (T4) , and on 5th day (T5).In S and IP groups, The rats were sacrificed at T1-3 and T5 , and the arcuate nucleus of the hypothalamus was removed for determination of NR2B mRNA expression (by real-time reverse transcriptase polymerase chain reaction) and NR2B and phosphorylated NR2B (p-NR2B) expression (by Western blot).Conclusion Compared with group S, the MWT was significantly decreased, and the TWL was shortened at T2-5 in IP, DMSO and Ro25-6981 groups, and the expression of p-NR2B was up-regulated at each time point (P＜0.05) , and no significant change was found in NR2B protein and mRNA expression in group IP (P＞0.05).Compared with group IP, the MWT was significantly increased, and the TWL was prolonged at T4in group Ro25-6981 (P＜0.05) , and no significant change was found in MWT and TWL at each time point in group DMSO (P＞0.05).Conclusion The activation of NR2B in the arcuate nucleus is involved in the development of IP in rats.

Objective To observe the effect of hydroxysafflor yellow A (HSYA) on the protection of blood brain barrier of cerebral ischemia mice, and explore the mechaniam. Methods Seventy-two C57/BL mice were divided into 6 groups: the sham operation group, the cerebral ischemia mice group, the TLR4 blocking group, the TLR4 blocking+cerebral ischemia mice group, the HSYA intervention+cerebral ischemia mice group, HSYA intervention+TLR4 blocking+cerebral ischemia mice group. Cerebral ischemia mice group were subjected to the middle cerebral artery occlusion (MCAO) model, TLR4 blocking was used, while TLR4 blocking was injected TLR4 antibody via right common carotid artery, and HSYA intervention group was injected 2 mg/kg HSYA by tail vein 0.5 h before cerebral ischemia. RT-PCR and western blot were applied to detect the mRNA and protein expression change of Wnt3a and β-catenin in each group. Results Compared with the cerebral ischemia mice group,the expression of TLR4 mRNA(1.63 ± 0.05,1.53 ± 0.04,1.84 ± 0.03 vs. 1.97 ± 0.05) significantly decreased (P<0.05 or P<0.01), the Wnt3a mRNA (0.56 ± 0.01, 0.58 ± 0.01, 0.50 ± 0.04 vs.0.42 ± 0.03),β-catenin mRNA(0.61 ± 0.03,0.74 ± 0.02,0.58 ± 0.04 vs.0.50 ± 0.03),Claudin-5 mRNA (0.54 ± 0.02, 0.58 ± 0.01, 0.47 ± 0.01 vs. 0.46 ± 0.02) mRNA significantly increased(P<0.05 or P<0.01), the protein expression of TLR4 (1.73 ± 0.05, 1.57 ± 0.03, 1.79 ± 0.08 vs. 1.89 ± 0.02) significantly decreased (P<0.05 or P<0.01), the protein expression of Wnt3a (0.67 ± 0.03, 0.74 ± 0.03, 0.57 ± 0.01 vs. 0.46 ± 0.01), Occludin(0.66 ± 0.02,0.73 ± 0.02,0.67 ± 0.01 vs.0.53 ± 0.01),Claudin-5(0.71 ± 0.01,0.73 ± 0.01,0.66 ± 0.01 vs. 0.64 ± 0.03) significantly increased (P<0.05 or P<0.01) in the TLR4 blocking+cerebral ischemia mice group, the HSYA intervention+cerebral ischemia mice group, HSYA intervention+TLR4 blocking+cerebral ischemia mice group. Conclusions TLR4 plays a critical regulatory role on the activation of Wnt3a and β-catenin in cerebral ischemic mice model. HSYA could intervene on the tight junction of cerebral ischemic brain through the intervention of Wnt3a and β-catenin, thus exerting the protection for cerebral ischemic brain.

Objective To discuss and evaluate the dosimetric characteristics of different plans implementing stereotactic radiotherapy(SRT)for intracranial tumors using Fixed and Iris collimators of CyberKnife VSI.Methods Twenty patients with intracranial tumors were selected and divided into group A with a small target volume(≤30 cm3)and group B with a large target volume(≥30 cm3). There were 10 patients in each group,and the prescribed dose to the target was 21 Gy in 3 fractions. For each patient, two treatment plans were designed using Fixed and Iris collimators. By analyzing the dosimetric parameters such as conformity index(CI),homogeneity index(HI), gradient index(GI), gradient score index(GSI), and organs at risk (OAR),the quality and efficiency of the plans were evaluated in order to discuss the beam characteristics for two sets of collimators. The difference was analyzed with the paired t-test. Results The mean time of Iris plan for delivering was significantly less than that of Fixed plan(group A:P=0.001;group B:P=0.000). In group B,the peripheral dose(20% and 10% of the prescribed dose)volumes of Fixed plan were significantly less than those of Iris plan(P=0.001 and 0.009). For OAR,D minof the visual pathway and D meanor D minof the eyeball in group B were significantly different between Fixed and Iris plans(all P<0.05), while in group A, only D minof the optic chiasm was significantly different between the two plans(P=0.043). For the other parameters of targets,there were no significant differences between Fixed and Iris plans in both groups(all P>0.05). Conclusions Apart from less treatment time in the Iris plan, there are no significant dosimetric differences between the two collimator plans of CyberKnife VSI in treating small intracranial tumor. For the large and complex tumor,although Iris plan meets the requirement for OAR dose constraints,its low-dose volumes are larger than those of Fixed plan. Further studies of the dosimetric characteristics in CyberKnife should be done.

Objective Toexploretheimagingfeaturesofextramedullarydisease(EMD)in multiplemyeloma(MM).Methods Theclinicalandimagingdataof17patientswithpathologicallydiagnosedMMcombinedwithEMDwereanalyzedretrospectively.Results EMDhadcertainpredilectionsites,Centralnervoussysteminvasion (6):meningealinvasion (3:1 multiple,2focal),spinalcanal invasion (1focal),thelefttemporalpoleinvasion(1focal),theleftsideforeheadinvasion(1focal);Headandneckinvasion(3:allfocal);Thoraxinvasion(8):pleuralinvasion(6:5 multiple,1focal),intrapulmonaryinvasion(1focal),anteriormediastinalinvasion(1focal);Subcutaneoussofttissueinvasion(5:allmultiple);Muscleinvasion(2focal);Lymphnodeinvasion (1 multiple).BothCTand MRI showedsofttissuenodulesormasses.ThevaluesofCTwereabout30~70HU,especiallyin30~45HU,whileMRIpresentedequal orslightlylowsignalonT1WI,equalorslightlyhighsignalonT2WI,andhighsignalinthesequenceofDWIcombinedwithmoderate toobviousenhancement,Generally,theboundaryofEMDwereclearandtheshapeoftheselesionswereregular,However,theinvasion tomuscleinsomelesionsshowedthepatternofinvasivegrowth.Conclusion EMDofmultiplemyelomamayhappenanywhere,and thepleural,meningesandsubcutaneoussofttissuesarethemostcommonlocation.CTandMRIcanshowtheEMDverywell.Thelocation, size,shapeandrelationshipwithsurroundingtissuesoftheselesionshavecertainreferencevaluesforthediagnosisanddifferential diagnosisofEMD.

Objective:To study the analgesic effect of α-cobratoxin (α-CbTX) on mice and its effect on protein kinase A (PKA) activity of spinal dorsal root ganglion (DRG) in mice.Methods:Healthy male ICR mice( n=102) were randomly divided into low-, medium-, and high-dose α-CbTX groups (1 mg/kg, 3 mg/kg, 9 mg/kg respectively, gavage, n=21), solvent control group (equivalent volume of 0.9% normal saline, gavage, n=21), morphine positive control group (3 mg/kg, intraperitoneal injection, n=6)or aspirin positive control group(300 mg/kg, gavage, n=12). The analgesic effect of α-CbTX was evaluated by hot plate test, acetic acid twisting test and formalin foot licking test. Formalin plantar injection was used to induce pain and then the L4-L6 DRG was taken 30 minutes later. The expression of PKA C-α in L4-L6 DRG of mice were detected by Western blot.SPSS 16.0 software was used for statistical analysis. Repeated measurement ANOVA was used to evaluate the hot plate experimental data, and one-way ANOVA was used for other experimental data. LSD- t test was used for further pairwise comparison. Results:In the hot plate test, the interaction between group and time of mice paw licking latency was significant ( F=8.902, P<0.05). At 0.5 h after administration, the paw licking latencies of α-CbTX medium-dose group ((11.83±1.47)s)and α-CbTX high-dose group (( 14.33±12.1)s) were both longer than that of solvent control group((8.17±0.75) s) ( t=4.461, 7.053, both P<0.05). The efficacy of α-CbTX medium dose group lasted until 1.5 h after administration (all P<0.05), and that of α-CbTX high dose group lasted until 2 h after administration(all P<0.05). In the acetic acid writhing test, the writhing times in the low-, medium- and high-dose α-CbTX group((34.50±3.62) times, (26.17±2.40) times, (13.83±3.76) times)) were significantly lower than that in solvent control group ((42.50±4.59) times) ( t=3.938, 8.040, 14.112, all P<0.05). In the period of the formalin test phase Ⅱ, the total licking time of α-CbTX low-, medium- and high-dose groups ((71.17±6.46) s), (54.67±6.41) s, (40.50±3.89)s) were significantly shorter than that of the solvent control group ((98.67±11.50) s)( t=6.950, 11.120, 14.700, all P<0.05). In the Western blot experiment, compared with solvent control group (0.22±0.01), the levels of PKA C-α in the DRG of mice in low-, medium- and high-dose α-CbTX groups ((0.31±0.02), (0.41±0.03), (0.44±0.02)) were up-regulated ( t=3.140, 6.471, 7.492, all P<0.05). Conclusion:α-CbTX has obvious analgesic effect, and its analgesic mechanism may be related to the activation of PKA.