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Objective To investigate the role of Wnt/β-catenin pathway in the glucocorticoids (GC)-mediated Alzheimer' s disease-like pathological changes in vitro.Methods Human embryonic kidney 293 (HEK293/wt) cells stably transfected with the longest human tau (tau441,HEK293/tau) and wild-type HEK293 cells were employed to study the role of GC.Cell viabilities of the two cell lines were examined by cell counting kit-8 ( CCK-8 ).Levels of phosphorylated tau ( p-T205 ) and dephosphorylated tau (Tau-1),β-catenin,phosphorylated β-catenin (p-β-catenin),glycogen synthase kinase-3β (GSK-3β),phosphorylated GSK-3β at Ser9 (ps9-GSK-3β) and Bcl-2 were determined by Western blotting.Results Treatment with 1 μmol/L GC for 48 h decreased the viability of HEK293/wt and HEK293/tau cells to 95.5％ ±3.2％ and 77.8％ ± 4.4％ (t =6.60,P ＜ 0.05 ).Moreover,GC treatment decreased the levels of ps9-GSK-3β,Tau-1,β-catenin and Bcl-2 to 47.8％ ± 10.4％,53.9％ ± 11.7％,50.9％ ±7.6％,48.4％ ±6.5％ of control groups ( t =7.01,3.86,7.09,7.30,all P ＜ 0.05 ),and increased the relative levels of pT205,p-β-catenin to 180.5％ ± 22.2％,201.3 ％ ± 27.6％ of control groups (t =5.51,5.27,both P ＜0.05) only in HEK293/tau cells.Finally,LiCI efficiently prevented the above effects of GC in HEK293/tau cells.Conclusion GC may trigger Alzheimer' s disease-like pathological changes by inhibiting the Wnt/β-catenin pathway and these pathological processes seem to specifically depend on the presence of human tau.
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In order to explore the role of acetylcholine in the pathogenesis of Parkinson's disease (PD), the changes in the concentration of acetylcholine (Ach) in the striatum, the apoptosis of substantia nigra cells, the ultrastructure and the changes of Nissl cells in rats during the morbidity of PD, and the corresponding behaviors in rats with PD were observed. Rat PD model was established by using the modified Thomas method. Eighty-one rats were randomly divided into normal control, sham operation and PD groups and their behavior features were observed at post-operative day (POD) 7, 14 and 21 as three subgroups (n=9 each). The concentration of Ach in the striatum was determined by using high-performance liquid chromatography. The apoptosis of substantia nigra cells was assayed by using TUNEL method. The ultrastructural changes in the substantia nigra were observed under the electron microscopy, and the survival of neurons in the substantia nigra area was examined by using Nissl staining. In PD group at POD 7 to 21, the damage in the substantia nigra area was gradually aggravated, the concentration of Ach, apoptosis rate and turns of rotation were gradually increased, and the number of Nissl cells was gradually reduced over the time as compared with the normal control and sham operation groups (all P<0.05). It was concluded that there exist dynamic changes in Ach concentration, ethology and apoptosis of the substantia nigra cells during the morbidity of PD, suggesting the contribution of apoptosis to the morbidity of PD, and critical role of Ach in the pathogenesis of PD.
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In order to explore the role of acetylcholine in the pathogenesis of Parkinson's disease (PD), the changes in the concentration of acetylcholine (Ach) in the striatum, the apoptosis of substantia nigra cells, the ultrastructure and the changes of Nissl cells in rats during the morbidity of PD, and the corresponding behaviors in rats with PD were observed. Rat PD model was established by using the modified Thomas method. Eighty-one rats were randomly divided into normal control, sham operation and PD groups and their behavior features were observed at post-operative day (POD) 7, 14 and 21 as three subgroups (n=9 each). The concentration of Ach in the striatum was determined by using high-performance liquid chromatography. The apoptosis of substantia nigra cells was assayed by using TUNEL method. The ultrastructural changes in the substantia nigra were observed under the electron microscopy, and the survival of neurons in the substantia nigra area was examined by using Nissl staining. In PD group at POD 7 to 21, the damage in the substantia nigra area was gradually aggravated, the concentration of Ach, apoptosis rate and turns of rotation were gradually increased, and the number of Nissl cells was gradually reduced over the time as compared with the normal control and sham operation groups (all P<0.05). It was concluded that there exist dynamic changes in Ach concentration, ethology and apoptosis of the substantia nigra cells during the morbidity of PD, suggesting the contribution of apoptosis to the morbidity of PD, and critical role of Ach in the pathogenesis of PD.
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Animais , Masculino , Ratos , Acetilcolina , Farmacologia , Corpo Estriado , Metabolismo , Patologia , Modelos Animais de Doenças , Doença de Parkinson , Metabolismo , Patologia , Ratos WistarRESUMO
ObjectiveTo investigate the initial symptoms, onset time and predisposing factors of acute cerebral haemorrhage in order to more effectively prevent cerebral haemorrhage. Methods 728 cases with acute cerebral haemorrhage were collected in recent 8 years. The initial symptoms,onset time and predisposing factors of all the patients were investigated. ResultsAcute cerebral haemorrhage in 582 caces (79.9%) occurred at time between 6am and 18pm and 146 cases (20%) at night. The main predisposing causes of acute cerebral haemorrhage were the playing of chess, cards and mahjong (16.1%), quarrels (12.8%), overwork (11.7%), alcoholism (7.2%) and so on. The most common initial symptom was hemianesthesia, accounting for 16.2%, others were vertigo (13.0%), headache ( 11.4%), amaurosis fugax ( 9.3%), speech ambiguity ( 8.6%) and transitory visual disturbance (7.9%).ConclusionsThe preventive measures of cerebral haemorrhage should be focused on daytime, especially in the morning. Sixteen locations where the cerebral haemorrhage is prone to happen should be paid more attention to by the patients with hypertension and the medical staff. Thirteen symptoms,such as hemianesthesia and vertigo,are the aurae of cerebral haemorrhage.
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The expression of the cytokines IL-2, IL-10, TNF-α and their roles in mice with herpes simplex viral encephalitis (HSE) were studied. By using semiquantitative reverse transcription polymerase chain reaction (RT-PCR), the expressions of IL-2, IL-10 and TNF-α mRNA in control group, HSE group and acyclovir (ACV)-treated group were detected and the pathological changes of brain were observed. It was found that after HSV1 infection, the cerebral lesions of haemorrhage and necrosis in mice were observed under the microscopy, and the levels of IL-2, IL-10 and TNF-α were increased remarkably. After treatment with ACV after HSV1 infection, the cerebral lesions in mice were improved, the level of IL-2 maintained stable, IL-10 was increased consistently, and TNF-α was decreased significantly as compared with those in HSE group. In acute HSE, many cytokines are upregulated, including IL-2, IL-10 and TNF-α to eliminate virus and TH1 type response is dominant. In convalescence, there is a shift in the cytokine expression profile from TH1 profile to TH2 profile and the shift can inhibit the overexpression of immune response in animals. ACV has remarkable effects in the treatment of HSE.
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The expression of the cytokines IL-2, IL-10, TNF-alpha and their roles in mice with herpes simplex viral encephalitis (HSE) were studied. By using semiquantitative reverse transcription polymerase chain reaction (RT-PCR), the expressions of IL-2, IL-10 and TNF-alpha mRNA in control group, HSE group and acyclovir (ACV)-treated group were detected and the pathological changes of brain were observed. It was found that after HSV1 infection, the cerebral lesions of haemorrhage and necrosis in mice were observed under the microscopy, and the levels of IL-2, IL-10 and TNF-alpha were increased remarkably. After treatment with ACV after HSV1 infection, the cerebral lesions in mice were improved, the level of IL-2 maintained stable, IL-10 was increased consistently, and TNF-alpha was decreased significantly as compared with those in HSE group. In acute HSE, many cytokines are upregulated, including IL-2, IL-10 and TNF-alpha to eliminate virus and TH1 type response is dominant. In convalescence, there is a shift in the cytokine expression profile from TH1 profile to TH2 profile and the shift can inhibit the overexpression of immune response in animals. ACV has remarkable effects in the treatment of HSE.
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BACKGROUND:It has been known that there exists a close relationship between leptin and obesity.Then,how are serum leptin and insulin related to obesity-related hypertension(ORH)? OBJECTIVE:To study the changes of serum leptin and insulin in elderly men with ORH,and investigate the relationship between leptin resistance and ORH in elderly men. DESIGN: Retrospective controlled study based on diagnosis. SETTINGS:The Department of Neurology and the Department of Nuclear Medicine,Affiliated Union Hospital of Tongji Medical College, Huazhong University of Science and Technology. PARTICIPANTS: Altogether 62 elderly male inpatients or outpatients aged from 60 to 82 years in the Affiliated Union Hospital of Tongji Medical College from October 2001 to June 2002 were selected.The diagnosis of hypertension was based on the diagnostic criteria established by the WHO in 1999:systolic blood pressure(SBP) ≥ 140 mm Hg(1 mm Hg=0.133 kPa) and(or) diastolic blood pressure(DBP) ≥ 90 mm Hg. INTERVENTIONS:The content of serum leptin and insulin in 41 hypertensive and 21 normotensive elderly men was determined by the method of radioimmunoassay. MAIN OUTCOME MEASURES:Blood pressure,body mass index (BMI),serum leptin and insulin level. RESULTS:The content of serum leptin and insulin in the hypertensive obese men were increased significantly by 1.8 μ g/L and 2.7 mIU/L,respectively,as compared with that in the normotensives (t=2.212,2.395,P< 0.01). The level of serum leptin and insulin in the ORH patients was increased by 2.7 μ g/L and 4.7 mIU/L,respectively,as compared with that of the normotensives with significant differences (t=3.348,5.113,all P< 0.001). The level of serum leptin in the hypertensive patients older than 70 years was significantly increased by 1.7 μ g/L compared with that of those aged 60 to 70 years(t=2.767,P< 0.05). CONCLUSION:Resistance to leptin and insulin exists in obesity-related hypertension of elderly men and leptin is closely associated with obesity-related hypertension.
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BACKGROUND: Microglial cells are prominently involved in certain neurologic diseases such as Parkinson disease and Alzeheimer disease. In vitro primary culture is commonly used in studies on the functions of microglia.However, these classical culture methods have some defects including complex procedures and low out-put.OBJECTIVE: To establish a simplified high-output primary culture of microglia.DESIGN: An explorative experiment with microglial cells as the single sample.SETTING: Department of Neurology, Affiliated Union Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The study was finished at the Central Laboratory of Union Hospital from April to October 2004. Microglial cells were obtained from 10 newborn(one day) male Kunming mice that were selected.METHODS: The author' s culture method was based on McCarthy method, we developed a new culture method and made some improvements,including the increased cell density for primary culture and nutritional deprivation. The microlglial cells were isolated with low-concentration trypsin-EDTA(ethylene diamine tetraacetic acid) digestion and immunochemically labeled with MAC-1 antibody, so as to measure the output and purity of microglia.MAIN OUTCOME MEASURES: ① Morphologic features of microglial cells, observed with inverted microscope; ② Purity and activity of microglia cultured with these two methods, were measured immunohistochemically.RESULTS: For microglia cultured with McCarthy method, the culture cycle was 20 days and the output was 2 × l05 cells per flask with a purity of 95% -97%. The new method shortened the culture cycle to 15 days and the output reached 1 × 106 cells per flask with a purity of 96-98%. Cell purity and activity had no significant difference between these two culture methods.CONCLUSION: The new method has a similar purity and activity with classical method; however, it may simplify procedures, shorten cycle, and increase output, and therefore can be a useful method for studies on microglia function and for nerve repair.
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AIM To study the effect of exogenous bFGF on the cell apoptosis and the expression of HSP 70 protein and p53 gene. METHODS The effect of exogenous bFGF on HSP 70 protein and gene expression was examined vial insitu hybridization and immunohistochemistry, at 0～72 h reperfusion after the middle cerebral artery (MCA) was occluded for 2 h in rats. Simutaneously, the distribution of apoptosis was observed. RESULTS The expression of HSP 70 protein elevated and the expression of p53 gene and cell apoptosis decreased in bFGF treated rats as compared with ischemia rats. (P<0.05, P<0.01). CONCLUSION Exogenous bFGF can supress the apoptosis and regulate its relative gene.
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The protective effect and mechanism of diazepam on ischemia neurons during cerebral ischemia and reperfusion were studied. Sixty-three Wistar rats were divided randomly into nine groups: control group (n=7), ischemia (is) groups including subgroups of is3h, is3-h/rep1-h, is3-h/rep2-h, is3-h/rep3-h(n=7 in each group), diazepam treated groups (10 mg/kg, i.p.), including subgroups of is3-h, is3-h/rep1-h, is3-h/rep2-h, is3-h/rep3-h (n=7 in each group) with Zea longa's animal model of middle cerebral artery occlusion. The comparison between the ischemia group and diazepam-treated group showed that diazepam could obviously decrease the production of glutamate, asparate, MDA and increase the synthesis and release of GABA, SOD and GSH-PX. It was concluded that diazepam exerted its protective effects on neurons through complex mechanisms of regulating the synthesis and release of excitotary/inhibitory amino acids and free radicals.
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The protective effect and mechanism of diazepam on ischemia neurons during cerebral ischemia and reperfusion were studied. Sixty-three Wistar rats were divided randomly into nine groups: control group (n=7), ischemia (is) groups including subgroups of is3h, is3-h/rep1-h, is3-h/rep2-h, is3-h/rep3-h(n=7 in each group), diazepam treated groups (10 mg/kg, i.p.), including subgroups of is3-h, is3-h/rep1-h, is3-h/rep2-h, is3-h/rep3-h (n=7 in each group) with Zea longa's animal model of middle cerebral artery occlusion. The comparison between the ischemia group and diazepam-treated group showed that diazepam could obviously decrease the production of glutamate, asparate, MDA and increase the synthesis and release of GABA, SOD and GSH-PX. It was concluded that diazepam exerted its protective effects on neurons through complex mechanisms of regulating the synthesis and release of excitotary/inhibitory amino acids and free radicals.
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AIM To study the effect of exogenous bFGF on the cell apoptosis and the expression of HSP 70 protein and p53 gene. METHODS The effect of exogenous bFGF on HSP 70 protein and gene expression was examined vial insitu hybridization and immunohistochemistry, at 0-72 h reperfusion after the middle cerebral artery (MCA) was occluded for 2 h in rats. Simutaneously, the distribution of apoptosis was observed. RESULTS The expression of HSP 70 protein elevated and the expression of p53 gene and cell apoptosis decreased in bFGF treated rats as compared with ischemia rats. (P