Objective To investigate the neurogranin expression in the rat neocortical and hippocampal regions after kainci acid(KA) induced limbic seizures.Methods 52 SD rats were divided randomly into KA group and normal control group.The animals of KA group were injucted KA to kindling the seizures and sacrificed at the end of 6 h,12 h,18 h,24 h,and 48 h respectively after seizures induced.By using fluorescent immunostain combined with confocal microscope,neurogranin expression and distribution were examined in the cerebral cortex and different regions of hippocampal.Western Blot technique was specially used to analyze the quantitative level of protein involved in the relate areas.Results Strong expression of neurogranin was present in cytoplasm of layers Ⅱand Ⅲ cortical,CA_1,CA_3 pyramidal cells and dentate gyrus granule cells of normal control groups.Rats after 18 h KA injection began to exhibit decreased expression of neurogranin in the cortex significantly((P

BACKGROUND: Apart from anticoagulation property and suppressing platelet congregation capability, tongxinluo preparation has been proved by traditional Chinese medicine to possess certain function for protecting endothelial cells.OBJECTIVE: To observe the influence of Chinese medicinal herb "tongxinluo" compound on adhesion molecule expression in brain ischemia-reperfusion (IR) animal model.DESIGN: Randomized and controlled experiment.SETTING: Department of Neurology, Changzheng Hospital Affiliated to the Second Military Medical University of Chinese PLA.MATERIALS: This experiment was conducted at the laboratory of the Department of Neurology, Shanghai Changzheng Hospital, between October 2002 and January 2003. Totally 25 male SD rats were randomized into sham-operation group of 5 rats, model group of 10 rats and tongxinluo group of 10 rats.METHODS: Middle cerebral artery was occluded using thread-bolt method to induce focal brain IR model in rats. In sham-operation group,nylon thread was placed around the external carotid artery approximating to the branch of internal carotid artery, and the other procedure was the same as that in model group. Rats in tongxinluo group were given tongxininfusion before IR for 1 consecutive week, which was replaced by physiological saline of the same dosage in model group and sham-operation group. Brain tissues were obtained under anesthesia condition and cut into slices; conventional HE staining, immunohistochemical and in situ hybridization staining were conducted.MAIN OUTCOME MEASURES:① The number of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1)positive microvessels following IR injury.② The number of ICAM-1 and VCAM-1 mRNA positive microvessels following IR injury.RESULTS:① In sham-operation group,ICAM-1,VCAM-1 protein andICAM-1 mRNA positive microvessels could not be observed in hemispheric cortex and basal ganglion at the operative side.② In model group,the positive expression of ICAM-1, VCAM-1 protein and ICAM-1 mRNA obviously increased at the ischemic side at 6-hour reperfusion following 2-hour ischemia.③ In tongxinluo medication group,the positive protein and mRNA-expressing microvessls were found remarkably reduced in number in ischemic side hemispheric cortex and basal ganglion [(10.42 ±1.98),(12.42±2.14)/HP; (8.54±2.00), (11.12±1.56)/HP] (P ＜ 0.05), but the positive VCAM-1 protein-expressing microvessels did not change remarkably (P ＞ 0.05).CONCLUSION: Tongxinluo can suppress ICAM-1 transcription and translation following rat brain IR, thus attenuating inflammatory injury induced by brain ischemia.

Background and purpose: Itch protein is an established regulator of T cell immune response thresholds, belong to a class of E3 ubiquitin-transferring enzymes, widely involve in the ubiquitination of several key signaling molecules, such as ZAP70, P85, VAV, PLC-γ, PKC-θ, etc, plays a critical role in tumor induced immu-nosuppression. Itch ligase activity regulate T-cell anergy and development of regulatory T cells in the periphery by modulating key components of T-cell receptor and transforming growth factor-βsignaling. Therefore, manipulation of Itch activities may provide the opportunities to develop future therapies for immune disorders such as autoimmunity and cancer. speciifc small interfering RNA(siRNA) was utilized to silence the expression of Itch gene of T-lymphocytes and investigate the cytotoxicity activity of transfected T lymphocytes against MFC stomach neoplasms cells in vitro. Methods:T lymphocytes were isolated from the spleen of 615 mice and transfected by speciifc siRNA to silence the expression of Itch gene, The expression of Itch protein were examined by Western bolt in each group;72 hours after transfection, The secretion level of IL-2, INF-γwere measured by enzyme-linked immunosorbent assay (ELISA). At the end, the cytotoxicity activity changes against MFC stomach neoplasms cells was compared between transfected T lym-phocytes, negative control and blank control in vitro. Results:Compared with control group, the expression rate of Itch protein of transfected T-lymphocytes was decreased to 16%after transfection 48 hours;72 hours after transfection, the secretion level of IL-2 in transfection group, negative control and blank control respectively were (1 891.96±141.91)pg/mL, (1 241.69±91.67)pg/mL and (1 175.03±89.14)pg/mL (P<0.001), the secretion level of INF-γin transfection group, negative control and blank control respectively were (958.33±75.46)pg/mL, (683.33±66.67)pg/mL and (691.72±68.72) pg/mL (P<0.05). Transfected T lymphocyte also showed more efifcient killing ability against MFC stomach neoplasms cells than negative control and blank control in vitro, the highest killing rate has reached (54.18±2.96)%. Conclusion:Silencing Itch gene can signiifcantly promoted the secretion level of IL-2, INF-γof mice T lymphocyte, enhanced the cytotoxicity activity of T lymphocyte against MFC stomach neoplasms cells in vitro.

Objective:To observe the quantitative and morphological changes of astrocytes in nucleus of solitary tract(NTS) after vagus nerve stimulation(VNS).Methods:Light and electron microscopy immunohistochemisty were used to examine the relationship between astrocytes and neurons by observing the expression of glial fibrillary acidic protein(GFAP),which is the specific protein for astrocytes. Results:Immunoreactivities of GFAP were significantly enhanced after VNS in NTS of rats. Meanwhile,the hypertrophy cell bodies and dark long processes could be seen under high power field. For electron microscopy,immunopositive GFAP astrocytes connected closely with the dendrites or axons of the neurons. The number of synapse increased remarkably after VNS compared with that of the control. Conclusion:Our data indicate that not only the neurons but also the astrocytes in the NTSs play a very important role in the process of the antiepileptic mechanism during VNS.

Objective To investigate the related brain areas and nucleus involved in the inhibition of vagus nerve stimulation (VNS) on epilepsy. Methods Using the kainic acid kindling epilepsy rats model,we observed the distribution of Fos positive neurons in the brain after VNS treatment combined with immunohistochemical method. Results VNS induced a significant increase in Fos immunoreactivity in the bilateral nucleus of solitary tract,the locus coeruleus,parabrachial nucleus,periaqueductal gray of midbrain,lateral habenular nucleus,paraventricular thalamic nucleus,rhomoid thalamic nucleus,paraventricular hypothalamic nucleus.Dense Fos immunoreactive staining was also seen in the central nucleus of amygdala,bed nucleus of stria terminalis,lateral septal nucleus and prepirifiorm cortex.Pretreatment with electric stimulation on cervical vagual nerve stem, c fos expressing of hippocampus formation,cingulate gyrus and frontal,parietal,temporal lobus significantly diminished after KA injection. Conclusion This finding may suggest that VNS activates various brain structure that could be involved in the regulation of seizures.

Objective To explore the protective effects of electro-acupuncture (EA) serum onβ-amyloid protein (Aβ) induced primary rat hippcampal neurons. Methods The rat models of Alzheimer's disease were established by intracerebral injection of Aβ1-40. After treated them with EA, the serum was harvested. Primary cultured hippocampal neurons were treated with Aβ25-35 to establish neuronal damage model in vitro, and divided into normal group, model group and EA serum group. The proliferation of neurons was detected by MTT test. Neuronal apoptosis was examined by TUNEL staining, and expression of cysteine aspartic acid proteases-3 (Caspase-3) was detected by immunocytochemical staining. Results MTT test showed that the cell viability was significantly decreased after treated with Aβ. While compared with the model group, cell proliferation of EA serum group was significantly enhanced (P<0.01). TUNEL staining showed that the number of apoptotic cells in EA serum group decreased significantly compared with the model group (P<0.01). After 48 h of Aβ treatment, Caspase-3 expression levels were significantly elevated. However, compared with the model group, the number of Caspase-3 positive cells in EA serum group was significantly reduced (P<0.01). Conclusion The EA serum could promote the proliferation of hippocampal neurons, reduce the expression of Caspase-3, counteract the neurotoxicity of β-amyloid protein, and reduce neuronal apoptosis.

Objective To observe the effects of electroacupuncture on the expressions of autophagy related protein Beclin-1 and apoptosis related protein p53 of hippocampus in rats;To explore the mechanism of electroacupuncture on Alzheimer's disease (AD).Methods The rats were randomly divided into the normal group, the sham-operation group, the model group, and the electroacupuncture treatment group. “Baihui” and “Yongquan” points were taken for electroacupuncture treatment and the treatment course was 7 days. The rats were treated once a day for 4 courses. Changes in morphology and number of Nissl positive cells were examined by Nissl staining in hippocampal CA1 regions. Expressions of Beclin-1 and p53 protein were determined by Western blot analysis.Results Number of Nissl positive cells in CA1 region of the model group was significantly less than that of normal group (P<0.01). After electroacupuncture treatment, number of pyramidal cells and expression of Nissl body significantly increased (P<0.05). Expression of Beclin-1 decreased, while expression of p53 increased in the hippocampus of the model group, compared with that in the normal group (P<0.05). However, electroacupuncture treatment could significantly upregulate the expression of Beclin-1 protein (P<0.01), but downregulate the level of p53 (P<0.05).Conclusion Electro-acupuncture treatment could fight against Aβ-induced neuronal apoptosis, and improve the morphological changes of AD’s hippocampus.

Through the construction practice of the optic nerve disease picture database,the paper discusses the system architecture,database field,data content,picture processing,organization and implementation,and other issues about the construction of the clinical case picture database,states and analyzes the operation effect,points out deficiencies,and provides reference for the construction of relevant picture databases.

Objective To observe the effect of enhanced external counterpulsation (EECP) on diabetic retinopathy (DR). Methods 179 patients who accepted EECP combined with medication were as group A and the other 190 patients who accepted medication only were as group B. Their visual acuity, fundus fluorescein angiography (FFA) and optical hemodynamics were compared. Results There was significant improvement in group A with visual acuity, FFA and optical hemodynamics (P<0.05), and the incidence of improvement was more in group A than in group B (P<0.05). Conclusion EECP is effective on diabetic retinopathy.

Objective: To evaluate the safety and anticoagulant efficacy of domestic bivalirudin injection during emergent percutaneous coronary intervention (PCI) in patients with ST-segment elevation myocardial infarction (STEMI).
Methods: A total of 75 STEMI patients were randomly divided into 2 groups according to anticoagulant used in emergent PCI procedure. Bivalirudin group, the patients received intravenous domestic bivalirudin, n=40 and Heparin group, n=35. The activated clotting time (ACT) was tested at pre-PCI, 5 minutes after medication, immediately after PCI, 30 minutes, 1 hour and 2 hours after medication respectively. The activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT) and ifbrinogen (FIB) level were measured at before medication and 6, 24, 72 hours after medication.
Results: All patients in Bivalirudin group had ACT>225s at 5min after medication as PCI requirement, while 1 patient in Heparin group could not reach the requirement and the extra dose was added. Both groups maintained ACT>225s during PCI procedure. Bivalirudin group had the lower ACT levels than those in Heparin group at 30 min, 1-and 2-hour after the medication, P<0.05. The post-PCI levels of APTT, PT, TT and FIB were similar between 2 groups, all P>0.05. The no-cardiac event surviving rate at 30 days after PCI in Bivalirudin group and in Heparin group were similar P>0.05 and the mild bleeding at 24 hours after PCI in Bivalirudin group was lower (0 vs 11.43)%, P<0.05.
Conclusion: Compared with heparin, domestic bivalirudin may take faster effect, with shorter half-life period for anticoagulation during emergent PCI procedure in STEMI patients.