AIM　To study the effects of Aβ25～35 and Apo E4 on neuronal intracellular free Ca2+(［Ca2+］i). METHODS　Hippocampal and cortical neurons suspension of newborn(0～3 days) SD rats was produced. After incubated with fura-2/AM,the neurons suspension was divided into four groups: control, Aβ25～35, Apo E4, Aβ25～35+Apo E4. Each groups ［Ca2+］i was measured using a RF-5000 dual wavelength spectrofluorometer after incubated with double distilled water, Aβ25～35, Apo E4, Aβ25～35+Apo E4 for 3 min, respectively. The neurons outocorrelation function(ACF) of the scattering light intersity was analyzed by the microscope quasi-elastic light scattering(MQLS) technique The frequency shift line width by ACF. The Γ can sympolize the cell menbrane flilidity. RESULTS　Both Aβ25～35 and Apo E4 could significantly enhance hippocampal and cortical neurons rest ［Ca2+］i, furthermore, the effect of 5 μmol*L-1 Aβ25～35 was higher than the effect of 1 μmol*L-1 Aβ25～35 (P<0.05), and they also amplified KCl-induced rise in ［Ca2+］i in hippocampal and cortical neurons(P<0.05). The interaction of Aβ25～35 and Apo E4 could also significantly enhance hippocampal and cortical neurons rest ［Ca2+］i andamplified KCl-induced rise in ［Ca2+］i in hippocampal and cortical neurons(P<0.05), but they had no synergic or additive effect.The frequency shift line widith Γ of both hippocampal and cortical neurons were decreased by both Aβ25-35 and ApoE4. CONCLUSION　Aβ25～35 and Apo E4 could enhance neuronal intracellular free Ca2+, amd decrease meirpma; ,e,brame f;iodotu. But their interaction had no synergic or additive effect. It suggested that the amplified effect of Aβ25～35 and Apo E4 on neuronal ［Ca2+］i and membane fluidity may be relative to their neurotoxity.

Through genetic recombination technique, the rat glial cell line-derived neurotrophic factor (rGDNF) cDNA was in-serted into polylinker site of retroviral vector pLXSN, to generate a recombinant plasmid pLXSN-GDNF as transfer vector. Therecombinant plasmid was verified with restriction analysis, PCR, dot blot hybridization and Southern blot hybridization. The re-sults showed that GDNF cDNA was cloned correctly into retroviral vector pLXSN, recombinant retroviral vector was construct-ed. It is concluded that the eukaryotic cell expression vector was constructed successfully for gene therapy of Parkinson's,Alzheimer's and other central nervous system diseases.

In order to investigate effect of basic fibroblast growth factor(bFGF) on the prolifiration and differentiation of theneural progenitor of embryonic hippocampus of rats in vitro, 25 ng/ml bFGF was employed into the serum-free medium of cultureof hippocampal neural cells of embryonic day 18 rats in the present study. The effect of bFGF on the viability of cells culturedwas detected by MTT colorimetric method and the effects of bFGF on proliferation and differentiation of hippocampal neural pro-genitors were analyzed qualitatively and quantitatively by means of immunochemistry, for the nestin, neurofilament, galactocere-broside and glial acidic fibrillary protein. The results showed that the OD value of experimental group was higher than that ofcontrol group by 1.5 and 1.8 times at 4 d and 8 d respectively. The quantitative analysis of each kinds of cells indicated that thenumber of neural progenitors, neurons and oligodendrocytes in experimental group were increased about 2 times as many as thatin control group, but no differences of astrocytes between the two groups at 4 d. However, the number of four kinds of cells aug-mented about 1.7 times at 8 d. The results of this study suggest that bFGF can not only promote the survival, proliferation, butalso facilitate the differentiation of neural progenitor of hippocampus to neurons and glial cells. To obtain more many purifiedneural progenitors in vitro, the embryonic day 18 is not an appropriate age. It is better to get younger embryo brain to culture and to add enough bFGF.

AIM To study the effects of A? 25～35 and Apo E4 on neuronal intracellular free Ca 2+ ([Ca 2+ ] i). METHODS Hippocampal and cortical neurons suspension of newborn(0～3 days) SD rats was produced. After incubated with fura 2/AM,the neurons suspension was divided into four groups: control, A? 25～35 , Apo E4, A? 25～35 +Apo E4. Each groups [Ca 2+ ] i was measured using a RF 5000 dual wavelength spectrofluorometer after incubated with double distilled water, A? 25～35 , Apo E4, A? 25～35 +Apo E4 for 3 min, respectively. The neurons outocorrelation function(ACF) of the scattering light intersity was analyzed by the microscope quasi elastic light scattering(MQLS) technique The frequency shift line width by ACF. The ? can sympolize the cell menbrane flilidity. RESULTS Both A? 25～35 and Apo E4 could significantly enhance hippocampal and cortical neurons rest [Ca 2+ ] i, furthermore, the effect of 5 ?mol?L -1 A? 25～35 was higher than the effect of 1 ?mol?L -1 A? 25～35 ( P

ObjectiveTo study the effect of facilitation techniques combined with acupuncture on movement function and activity of daily living(ADL) of the stroke patients.Methods200 patients with stroke were divided randomly into 4 groups,group A for facilitation techniques with acupuncture,group B for facilitation techniques only,group C for acupuncture therapy only,and group D for control.The recovery grades of the Brunnstrom and Functional Independent Measure(FIM) were applied to assess movement function and ADL before and after treatment.ResultsThe number of patients whose limbs recoveried up to Brunnstrom 4 in group A、B、C after treatment was higher than that in group D as while as the scores of FIM(P<0.05).The FIM scores and the number of patients whose whose limbs recoveried grades(>4 grades) of the Brunnstrom in group B and C were no difference(P>0.05),The FIM scores and the number of patients whose recovery grades(>4 grades) of the Brunnstrom in group A were significant higher than that in group B and C (P<0. 05).ConclusionsFacilitation techniques combined with acupuncture can improve the movement function and ADL of the stroke patients.

BACKGROUND: It has been demonstrated that amyloid-beta 42 protein (Aβ42) immunization in transgenic mouse models of Alzheimer disease(AD)can induce specific Aβ42 antibody, clear Aβ from the brain, and thereby improve spatial learning and memory. It has been a promising treatment strategy for AD.OBJECTIVE: To explore the effect of Aβ42 and its subunit vaccines immunization on spatial learning and memory of APPSWE transgenic mice.DESIGN: A randomized controlled experiment with mice as subjects.SETTING: The brain research laboratory of the anatomy department in a the medical college of a univeristy.MATERIALS: The experiment was conducted in the Experimental Animal Center and the Anatomy Department of Sun Yat-sen University from April 2003 to February 2004. Thirty-two APPSWE transgenic mice of 5 months old were bought from Taconic Company, USA. The second generation of mice were successfully reproduced in the Anatomy Department. These mice were randomly assigned into four groups: control group, Aβ42 group, Aβ1-15group, and Aβ36-42 group. Each group contained 8 in each group.INTERVENTIONS: Aβ42 and its subunits combined with MF59 adjuvant were subcutaneously injected for fundamental immunity and then applied in nasal mucosa for intensified immunization. The immunization period was 8 months. Y-maze was used for behavior test before immunization and Morris water maze was used after immunization.MAIN OUTCOME MEASURES: Spatial learning and memory, mean escape latency, times of passing through the platform point, swimming distance percentage of the first quadrant, and swimming distance percentage of the 20% marginal area.RESULTS: The correct reaction times in Y-maze behavior test were 7.50 ±0. 81, 7.06 ±0.71, 7.19 ±0.91, and 7.50 ±0.86 respectively in the control, Aβ42, Aβ1-15, Aβ36-42 groups and there was no significant difference ( P ＞ 0. 05) . After immunization, the mean escape latencies in 8 units of localized navigation test were(67.3 ±2. 8) s, (23.6 ± 1.6) s, (26.4 ±2.0) s,and (36.5 ± 2.2) s. The results in three experiment groups were different from that in control group and there was no difference between the three experiment groups ( P ＞ 0. 05 ) . The mean times of passing through the platform point in the 4 groups were 0.71 ±0.29, 8.14 ± 1.37, 7.28 ± 1.34,and 3.29 ± 0. 67. Swimming distance percentage of the first quadrant in the4 groups were(24.3 ±2.9)%, (50.6±11.6)%, (49.9±9.3) %,and(35.4±7.0)% and the swimming distance percentages of 20%marginal area were (46.4 ± 7.3 ) %, ( 11.6 ± 3.9) %, ( 14.4 ± 2. 6) %, and (25.8 ± 3.3)%. The mice in three experiment groups showed increase in the times of passing through platform point, swimming distance percentage of the first quadrant, and decrease in distance percentage of 20% marginal area compared with control group. The results in three experiment groups were no significantly different( P ＜ 0. 05).CONCLUSION: Immunization with A342 and its subunits can effectively ameliorate impairment of spatial learning and memory in APPSWE transgenic mice.

OBJECTIVE: In recent years, available evidence from basic and clinical research on Alzheimer disease(AD) suggests that oxidation stress is involved in the occurrence and development of AD, and that antioxidant treatment can improve the intelligence of patients with AD and delay age-dependant cognitive dysfunction. Although results of basic and clinical research on the therapeutic effects of antioxidants on AD are inconsistent, a large number of available data suggest that these studies are of significance. Basic pharmacological studies on natural antioxidant TA99 series indicate that they are promising novel drugs for AD. Thereby, this study made a review of their experimental basis in the treatment of AD and existing problems.DATA SOURCES: Related articles published between January 1991 and December 2004 were searched by the computer in Medline database with such key words as Alzheimer disease, antioxidant, Ginkgo biloba extract, TA9901,acetylcholine, and senescence-accelerated mouse in different combinations and with the language limited to English. Meanwhile, related articles were alsosearched in CDMA \Wanfang database with the same key words in Chinese.STUDY SELECTION: Literature involving intervention group and control group were screened in the first trial, and then non-randomized trials were excluded and the rest were searched for the full text.DATA EXTRACTION: Of the 24 basic and clinical randomized and non-randomized trials on antioxidants in the treatment of AD collected, 17 accorded with the inclusion criteria and the other 7 were excluded.DATA SYNTHESIS: Intervention in the 17 trials emphasized the pathogenesis of AD from amyloid β proterin(Aβ) synthesis, gathering to senile plaque formation, and the enhancement of Aβ gathering and neuronal apoptosis by peroxidative injuries of free radicals. Both in vitro and in vivo studies were conducted: the effect of Aβ on neurons of different regions was observed with cell culture; transmission electromicroscope and sulfrin T (Th-T) fluorescence assay, Fuliye-transform infrared(FT-IR) spectrum apparatus, electron magnetic resonance(EPR), and round spectrum were used to detect the inhibitory effect of TA99 series on Aβ gathering and fibroplasia in vitro, as well as the influence on Aβ gathering in vivo. Senescence accelerated mouse (SAM) -P/8 was adopted to establish AD model and behavioral studies such as Morris water maze were used to investigate their effect on learning and memory. Meanwhile, the clearance of intracerebral amyloid granular deposition due to TA99 was also observed with hexamic argent staining. The effects of TA series on Aβ target and possible mechanism were fully revealed, and basic pre-clinical data collection was almost completed.CONCLUSION: TA9901 plant extractions have been proved to inhibit Aβ gathering and fibrosis, and improve learning and memory of SAM-P/8 rats. Moreover, TA9902 prepared by TA9901 combined with EGb761, another synergic herb, has an obvious anti-neurotoxic effect by inhibiting Aβ gathering, fibrosis and secondary structural changes. Further pharmacological research is needed and will have a promising prospect.

Objective To evaluate left ventricular diastolic and systolic function of the dogs during different status of acute myocardial ischemia with velocity vector imaging (VVI). Methods Six healthy mongrel dogs were enrolled. Home-made artery constrictor was placed in the proximal coronary artery after opening the chest of dogs to produced different grades of coronary artery stenosis. Mean of peak diastolic velocity (Em), peak strain rate of diastole (Esr), mean of peak systolic velocity (Sm), peak strain rate of systole (Ssr), ejection fraction (EF) and left ventricular pressure were measured. Results There was good linear correlation between Em and Esr by VVI and left ventricular end diastolic pressure (LVEDP) (r=-0.834, P＜0.001 and r=-0.703, P＜0.001, respectively). Em and LVEDP had better correlation than that between Esr and LVEDP (P=0.032). Sm, Ssr or EF had significant correlation with +dp/dt_(max) (r=0.883, P＜0.001, r=0.772, P＜0.001 and r=0.647, P＜0.001, respectively). Significant difference was observed for correlation coefficient between Em and Esr and EF (P＜0.001). Conclusion Em and Sm are sensitive echocardiography indexes to evaluate left ventricular diastolic and systolic function, and are better than EF, Esr and Ssr.

Objective To evaluate the effect of remote limb ischemic postconditioning on the level of adiponectin during myocardial ischemia-reperfusion (I/R) in rats.Methods Fifty-seven male Sprague-Dawley rats,aged 8 weeks,weighing 250-350 g,were randomly divided into 3 groups (n =19 each):sham operation group (group S); group I/R; remote limb ischemic postconditioning group (group RLIP).Myocardial ischemia was induced by occlusion of anterior descending branch of left coronary artery for 30 min followed by 180 min reperfusion in urethane-anesthetized rats.In group S,the anterior descending branch was only exposed but not ligated.The animals underwent 10 min ischemia of bilateral hind limbs starting from 20 min of occlusion of the anterior descending branch,followed by reperfusion in RLIP group.The changes in S-T segment were recorded at 30,60,120 and 180 min of reperfusion.At 180 min of reperfusion,the blood samples were taken from the jugular vein for determination of serum levels of creatine kinase MB (CK-MB) and cardiac troponin Ⅰ (cTnⅠ).The animals were then sacrificed and hearts were removed for measurement of myocardial infarct size (IS) and adiponectin contents in myocardial tissues (by ELISA).Results Compared with group S,S-T segment was significantly elevated at each time point,myocardial IS,and serum cTnⅠ and CK-MB levels were increased,and adiponectin contents were decreased in I/R and RLIP groups.Compared with group I/R,elevation of S-T segment was significantly decreased at each time point,myocardial IS,and serum cTnⅠ and CK-MB levels were decreased,and adiponectin contents were increased in RLIP group.Conclusion The mechanism by which remote limb ischemic postconditioning reduces myocardial I/R injury is related to increased level of adiponectin in the myocardium of rats.

Objective To summarize the therapeutic results and experience of the interventional managements for pseudoaneurysms occurred after surgery. Methods Five pseudoaneurysms with different location that occurred after surgery in five patients were treated with different interventional managements. One patient suffered from a ruptured pseudoaneurysm of left common iliac artery, which was treated by obstructing the diseased artery with balloon via the abdominal aorta followed by the replacement of vascular prostheses. One patient had a pseudoaneurysm of right subclavian artery and endovascular covered stent was employed to isolate it. The remaining three patients were affected by pseudoaneurysm of terminal arteries and intraarterial embolization with gelfoam and/or steel coils was camed out. Results Complete closing of the pseudoaneurysm was obtained in all five patients and no therapy-related complications occurred. Conclusion Pseudoaneurysms occurred after surgery can be effectively treated with different interventional managements, it is worth popularizing this technique in clinical practice.