Objective To evaluate the changes of GRP78 expression in the Spinal Cord of Ischemia/Reperfusion Injuried Rat.Methods Fifty five adult Wistar rats(250～300 g)were randomly divided into 2 groups as control group(n=5)and operation group(n=50).The spinal cord SCII model was established.The expression of GRP78 was detected in spinal cord tissue through immunohistochemistry(IHC)and Western blot analysis,and the neuronal apoptosis was detected through terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)methods.Results The GRP78 expression was increased at 30 min of reperfusion,and peaked at 7 h,but began to decline at 11 h post-reperfusion,and reduced significantly at 23 h.The number of GRP78 positive neurons at 0.5,3,7,11 and 23 h groups was(19.4?1.34),(42.6?2.30),(82.4?2.07),(40.0?1.58)and(18.8?0.83),respectively and significantly higher than that of control group(P

OBJECTIVE:To establish an RP-HPLC analysis of Carthamin in mice and to study its pharmacokinetics.METHODS:The serum concentration of Carthamin was determined by RP-HPLC.The blood concentration-time curve was established and the main pharmacokinetic parameters were computed.RESULTS:The linear range of Carthamin was 0.558～55.8 ?g?L-1(r=0.999 2),with the lowest limit of detection at 0.005 ?g?L-1Carthamin in vivo assumed two-compartment model and rapid absorption.CONCLUSION:The proposed method is simple,sensitive and reproducible,and it met the standard for pharmacokinetic study.

Objective To observe the effects of electro-acupuncture on the expression and inhibition of DNA binding protein 2 (Id2) and myelin basic protein (MBP),and to explore the mechanism of remyelinization after compressive spinal cord injury (CSCI) in rats.Methods Fifty-four SD rats were randomly divided into a control group and a treatment group,and each was further subdivided into 3 time point subgroups:3,7 and 14 days.There were 9 rats in each subgroup.The CSCI models were made with a self-designed method.The acupuncture points Jiaji (EX-B2),bilateral Zusanli (ST36) and Taixi (KI3) were selected for treatment.Electro-acupuncture (continuous wave,2 Hz,1.5 V)was applied to the bilateral Zusanli (ST36) and Taixi (KI3) points.The control group received the injury but no treatment.The changes in the ultrastucture of the nerve fibers＇ white matter were de-termined by transmission electron microscopy (TEM).The alterations in the expression of MBP and Id2 were observed by double labeled immunofluorescence and Western blotting on the 3rd,7th and 14th day after the injury.Results TEM showed that the myelin sheaths in the control group had degenerated,swollen,and even broken down after CSCI.Changes to the myelin sheaths in the treatment group were milder than those in the control group.The immunofluorescence results showed the amount of Id2-immunoreactive oligodendrocytes in the control group to be (20 ±2) on the 3rd day after CSCI,becoming (16 ± 1) on the 14th day.The differences among the 3 control subgroups were not statistically significant.The amount of Id2-immunoreactive oligodendrocytes in the treatment group was (13 ± 1) on the 3rd day,reaching a minimum the 14th day.The differences among the 3 treatment groups were statistically significant.The differences compared with the control group at the same time points were also statistically significant.Western blotting showed that the expression of Id2 in the contrast and treatment groups was (1.12 ±0.12) and (0.67 ±0.01) respectively on the 3rd day after CSCI,and both decreased with time.The expression of Id2 in both groups reached their minima ((0.86 ±0.02) and (0.25 ±0.01) respectively) on the 14th day.The difference between the treatment groups and the contrast group was statistically significant at each time point.The expression of MBP in the contrast and treatment groups at day 3 was (0.44 ± 0.02) and (0.67 ± 0.04) respectively,and these increased with time.The expression of MBP in both groups peaked at the 14th day (at (0.95 ± 0.04) and (1.74 ± 0.09) respectively).These differences were again statistically significant.Conclusion Electro-acupuncture can regulate the expression of Id2 and MBP after CSCI.The down-regulation of Id2 which controls MBP negatively and the up-regulation of MBP may contribute to remyelination in the injured spinal cord.

Objective To investigate the changes of dopamine-?-hydroxylase(DBH) and activator protein 2-?(AP-2?) expression in spinal cord under the condition of stress or pain stimulation,so as to explore the mechanism for changes of noradrenergic(NA) neurons in the spinal cord of rat pain model.Methods Immunohistochemical staining,double immunofluorescent staining,Western blotting and computing-image analysis system were used to detect the changes of DBH/AP-2? expression in the spinal cord of formalin-induced rat pain model.Results A small number of DBH-positive neurons were sparsely distributed in the ventral horn of the normal spinal cord,while in the formalin-treated group,much more darkly-stained DBH-positive neurons appeared primarily in the ventral horn,intermediate zone,and the dorsal horn,which reached the highest level on day 3 after formalin-injection.The grey value and number of DBH-positive neurons on day 7 after injection began to decrease,but still higher than that in the control group.Compared with control group,the number of noradrenergic neurons in spinal cord of formalin-treated rat was increased significantly,which was also confirmed by Western blotting.Double immunofluorescent staining showed that DBH and AP-2? co-existed in the cells of the spinal cord.The changes of AP-2? expression were similarly to that of DBH in the spinal cord of rat pain model.Conclusion Our results indicated that some non-noradrenergic neurons with different chemical properties might convert into noradrenergic neurons under pain stimulation;noradrenaline may be involved in the formalin-induced pain and behavior regulation;As one of transcription factors,AP-2? may promote the DBH synthesis.

BACKGROUND: Aquaporin-4 may be one of the candidates for inducing the brain edema in ischemic stroke, however, it still has not been reported whether aquaporin-4 is involved in the formation of brain edema after intracerebral hemorrhage (ICH).OBJECTIVE: To observe the expression of aquaporin-4 of cerebral tissue in the pathologic course of hemorrhagic cerebral edema in rats, and investigate itsrelationship with the formation of brain edema following ICH.DESIGN: A randomized controlled experimental study.SETTING: Department of Neurology, People's Hospital of Guangxi Zhuang Autonomous Region; Research Room of Neurobiology, Chongqing University of Medical Sciences.MATERIALS: This experiment was carried out in the Research Room of Neurobiology, Chongqing University of Medical Sciences between April and October 2003. Totally 120 adult healthy male Wistar rats, weighing 250-300 g, were provided by the Experimental Animal Center of Chongqing University of Medical Sciences.METHODS: The experiment had four parts, each part (n=30) was divided into the control group (n=5) and operation group (n=25), and then the latter was subdivided into 6-hour, 1, 3, 5 and 7-day following ICH groups with 5rats in each group. In the operation group, ICH models were established by infusing collagenase into left caudate nucleus of rats unilaterally. The operative process in the control group was the same as that in the operation group except for infusing collagenase. Standards for successful model establishment: the paralytic forelimb flexed under abdomen after withdrawal,and the normal forelimb extended to the ground. ① The expression of aquaporin-4 was detected with immunohistochemistry. ② The expression of aquaporin-4 mRNA was assessed with in situ hybridizution. ③ The expression of aquaporin-4 mRNA was assessed with reverse transcription polymerase chain reaction (RT-PCR). ④ The pathological changes of hemorrhagic edema were observed under transmission electron microscope.MAIN OUTCOME MEASURES: ① expression of aquaporin-4 mRNAand protein; ② pathological changes of hemorrhagic edema.RESULTS: The model establishments were all successful in the operation group, and all the rats were involved in the analysis of results without deletion. ① Comparison of the expression of aquaporin-4 mRNA and protein among the groups. There was no significant change in the control group. As compared with the control group, there was a significant increase of the expressions of aquaporin-4 mRNA and protein in the edematous tissue at 6 hours following ICH, and then reached the peaks at 3 days, and the expression of aquaporin-4 was still significantly higher than that in the control group at one week following ICH (t=12.65, P ＜ 0.01). ② The corresponding sequential pathological changes in the edematous tissue of rats in the operation group: There was a gradual increase of intracellular edema within 1-3 days following ICH, and then the brain edema became aggravated at 3 days, an emergence of vasogenic edema and local edema tissue degeneration and necrosis were observed.CONCLUSION: The increased expression of aquaporin-4 was obviously enhanced following ICH, suggesting that aquaporin-4 may play an important role in the pathological course of hemorrhagic brain edema, and inhibition of aquaporin-4 expression may be an effective pathway to prevent and treat brain edema.

This paper proposed the idea of building the applied anatomical experiment and research platform for surgical postgraduates with professional degree,establishing double-tutorial system and applying applied anatomical teaching in basis course learning,clinical skill training and research capacity cultivating after analyzing the reasons of poor applied anatomical background of surgical postgraduates with professional degree.These ideas were intended to improve the cultivation quality.

Objective To investigate correlation between demyelination and caspase-12 expression alteration after compressed spinal cord injury (CSCI) so as to discuss mechanism of demyelinating lesion after CSCI.Methods Seventy-five adult SD rats were randomly divided into five groups,ie,normal group,control group,compression 1 d,3 d and 7 d groups,with 15 rats per group.Models of spinal cord compression were established with a self-made device.Ultrastructure of the demyelinated nerve fibers was observed by electronic microscope and oligodendrocyte apoptosis was detected by TUNEL staining and double labeling immunofluorescence.Immunoblotting was used to defect caspase-12 that was related to cell apoptosis.Results Demyelination of nerve fiber occurred after CSCI and was aggravated with time.Apoptosis of oligodendrocytes was found after CSCI,and showed significant difference between compression 7 d group and normal group (P ＜ 0.05).Caspase-12 was also upregulated with extension of compression time.Conclusion Caspase-12 mediating oligodendrocyte apoptosis is one of the mechanisms of nerve fiber demyelination after CSCI.

Objective To investigate the role of demyelination and the alteration of chondrotin sulfate proteoglycan (CSPG,NG2) expression after compression injury of the spinal cord (CSCI).Methods Seventy-five adult Sprague-Dawley rats were randomly divided into a normal group,a sham-operation group,a CSCI 1 day group,a CSCI 3 day group,and a CSCI 7 day group.There were 15 rats in each group.The injuries in the CSCI groups were inflicted using a technique devised in our laboratory.Basso-Beattie-Bresnahan (BBB) neurological function assessment was used to assess the rats' motor function,osmic acid staining and transmission electronic microscopy (TEM)were used to observe any pathological changes of myelinated nerve fibers in the white matter at 1,3 and 7 days after CSCI.The amount of myelinated nerve fibers in the posterior funiculus of the spinal cord and the ratio of myelin sheath thickness to axon diameter (the G-ratio) were calculated.Any alteration in NG2 expression was observed by Western blotting.Results The average neurological function assessment scores in the CSCI groups were (1.23 ±0.45),(0.65 ± 0.35) and (0.00 ± 0.00) respectively.Compared with the normal group (21.00 ± 0.00) and the sham operation group (21.00 ± 0.00),the differences were all statistically significant.The rats' motor function deteriorated gradually with time after the CSCI.Osmic acid staining showed that the white matter was intact in the normal and sham groups.After being compressed the myelinated nerve fibers became swollen,degenerated and broke down.The amount of myelinated nerve fibers in the normal group,the sham operation group and the three CSCI groups was (2771 ± 108),(2675 ± 199),(2403 ± 161),(1708 ± 70) and (8 10 ± 95) respectively.The amount of myelinated nerve fibers decreased in the CSCI groups and reached a minimum on the 7th day.The difference was statistically significant.The TEM quantity analysis showed that the G-ratios in the normal,sham operation,and CSCI 1 day,3 day and7 day groups were (18.10±0.4),(17.70±1.0),(6.69 ±0.8),(5.73 ±0.4) and (4.95 ±0.5) respectively.Compared with the normal and sham operation groups,the G-ratios in the 3 CSCI groups were lower and reached their minimum on the 7th day after injury.The difference was statistically significant.TEM observation showed that the axons and myelin sheaths were intact in the normal and sham groups.After CSCI the axons became swollen and cell organelles in the axoplasm degenerated and decreased.The layers of myelin sheath shrank,folded and even wrinkled,which had an onion-like appearance.The oligodendrocytes exhibited chromatin condensation.Macrophages showed infiltration.Western blotting showed that the expression of NG2 in the CSCI groups reached a maximum on the 1st day after injury and then decreased with time.The expression of NG2 in the CSCI groups was higher than in the normal and sham groups,and the difference was statistically significant.Conclusion Demyelination occurs after CSCI-the amount of myelinated nerve fibers decreases and neurological deficits increase with time.The expression of NG2 was associated with changes in the myelin sheaths after CSCI and contributed to recovery of the myelin sheath through proliferation and differentiation to oligodendrocytes and perhaps other kinds of cells.

ObjectiveTo evaluate the therapeutic efficacy and prognostic factors of intensity-modulated radiotherapy combined with chemotherapy for early-stage nasopharyngeal carcinoma patients in northwest China. MethodsFrom January 2006 to December 2009,58 patients with early-stage nasopharyngeal carcinoma were treated with IMRT in Xijing hospital,the clinical data were analyzed retrospectively.Survival rates was calculated by the Kaplan-Meier method and the differences was compared by the Logrank test.Univariate analysis method was use to identify all significant factors.ResultsThe follow-up rate was 100％.The follow-up time of 46 patients was more than 3 years.The 1-,2 and 3-year survival were 98％,94％ and 91％,respectively.The 3-year overall survival (OS),local recurrence-free survival (LRFS),distant metastasis-free surv ival (DMFS) for T1N0-1,T2N0 and T2N1 stage were 100％,100％,100％ and 74 ％,81％,87 ％,respectively ( x2 =5.74,P =0.01 ; x2 =4.95,P =0.03 ; x2 =4.24,P=0.04).The 3-year OS,LRFS,DMFS for IMRT combined with chemotherapy and IMRT alone were 100％,100％,100％ and 85％,85％,88％ respectively ( x2 =4.02,P =0.04; x2 =4.12,P =0.03 ; x2 =4.84,P =0.02).In T2N1 stage,IMRT combined with chemotherapy and IMRT alone were 100％,100％,100％ and 79％,79％,80％ respectively (x2 =5.28,P =0.03 ;x2 =4.84,P =0.04;x2 =4.72,P =0.04).In univariate analysis,N stage,clinical stage,IMRT combined with chemotherapy were significantly associated with the survival ( x2 =5.39,P =0.02 ; x2 =5.74,P =0.01 ; x2 =4.02,P =0.04).Conclusions In all early-stage nasopharyngeal carcinoma,T2N1 stage is a sub-group of high risk of distant metastasis.Combination of IMRT and chemotherapy may improve the LRFS,DMFS and OS in those patients.