This study is to investigate the effect of baicalin (BL) against oxidative injury stress of SH-SY5Y cells induced by H2O2 and the possible mechanism. SH-SY5Y cells were pre-incubated with baicalin (25, 50, and 100 micromol x L(-1)) for 12 h prior to exposure to H2O2 (150 micromol x L(-1)) for 24 h. The viability of SH-SY5Y cells was measured by MTT assay. The contents of LDH and NO were determined. The percentage of apoptotic cells was assessed by flow cytometry (FCM). The content of Caspase-3 was tested by immunofluorescence histochemical method. BL at 50 and 100 micromol x L(-1) separately increased the cell viability and up-regulated SIRT1, reduced the contents of LDH, NO, Caspase-3 and the apoptotic percentage of SH-SY5Y cells. This study results suggest that baicalin could inhibit the H2O2-induced neuronal apoptosis. The further mechanism studies show that baicalin inhibit apoptosis via reducing Caspase-3 expression and up-regulating SIRT1.

Objective To investigate the association between the genetic polymorphisms of vitamin D receptor(VDR)and bone mineral density (BMD) in elderly male patients with chronic obstructive pulmonary diseases. Methods Pulmonary function, body mass index(BMI), serum calcium, serum phosphorus and bone glaprotein were determined respectively in COPD group and control group. In the two groups the VDR gene polymorphisms were analyzed by polymerase chain reaction and restriction fragment length polymorphism (PCR RFLP) and BMD were investigated by dual energy X ray bone mineral density. Results There were differences in BMD, BMI and bone glaprotein between the two groups. The BMI, bone glaprotein and BMD of femur neck in COPD group were(22 65?3 18)kg/m 2 ,(2 50?0 57) ?g/L and(0 75?0 13)g/cm 2 respectively, which were more decreased than those in control group: BMI (24 86?3 68)kg/m 2 , bone glaprotein (2 87?0 61) ?g/L and BMD of femur neck (0 86?0 12) g/cm 2 ( P

Objective To study the neuropathological characteristics of late-onset Alzheimer' s disease (LOAD) in Chinese people, to ensure correct diagnosis of LOAD.Methods Choosing cerebral cortex of temporal layer of 8 cases of LOAD and 5 cases of age-matched normal control group by autopsy.Histopathologlc diagnosis was established in all these 13 cases.Cerebral cortex were taken from temporal layer in 13-101 hours after death and were fixed with 40 g/L paraformaldehyde, followed by paraffin-embedding and serial sectioning with 6 μm thickness.Brain tissue was analyzed neuropatholically by using immunohistochemical staining for β-amyloid (Aβ) and AT8 on these cases.Positive distribution of temporal layer was observed under light microscope.Results The results of immunohistochemical stainings of Aβ and AT8 were positive in all of LOAD.Aβ immunoreactant located in the cerebral cortex.The diffuse plaques, primitive plaques and burn-out plaques of senile plaques were displayed clearly by immunohistochcmical stainings of Aβ.AT8 immunoreactants showed neurofibrillary tangles, neuropil thread and senile plaques in nerve cell of cerebral cortex in different degree respectively.The positive rate Aβ and AT8 were both 8/8 by semiquantitative analysis in AD group.As the normal aging control group, which was 0 and 1/5 respectively.There was significant difference of the positive rate Aβ and AT8 in two groups(χ2 = 13.000,P=0.001; χ2=9.244,P=0.007).Conclusions Sensitive immunnhistochemical technique was significant to display senile plaques and neurofibrillary tangles.The findings demonstrate that immunohistochemistry staining of Aβ and AT8 can display senile plaques and neurofibrillary tangles clearly.The connection of the 2 different methods might improve diagnose accordance rate of AD.

Purpose To study the differences of shear wave elasticity and the content of collagen fiber between benign breast lesions and malignant breast lesions, and to analyze the correlation between shear wave elasticity and the content of collagen fiber. Materials and Methods 106 patients with 116 breast lesions who were referred to PLA General Hospital for ultrasound-guided biopsy or surgery underwent shear wave elasticity examination, the biopsy specimen underwent Van Gieson (VG) dye and Image-Pro Plus 5.1 software was used to quantitatively analyze the content of collagen fiber. Results Malignant lesions exhibited signiifcantly higher max elasticity, mean elasticity, and elasticity ratio between lesions and surrounding parenchyma (140.43±70.16) kPa, (63.11±33.68) kPa, 3.49±1.95 than benign lesions (54.64±48.53) kPa, (34.52±25.23) kPa, 2.25±1.48 (t=5.329, 4.382 and 4.487, P<0.01). The content of collagen fiber of malignant lesions was signiifcantly higher than that of benign lesions (t=8.437, P<0.01). There was a positive correlation between max elasticity and the content of ifber collagen (r=0.746, P<0.05). Conclusion The elasticity of breast lesions has a close correlation with the content of collagen ifber, and collagen ifber might play an important role in the development of breast carcinoma.

Objective To explore histopathological features in the nigrostriatal tissues of Parkinson’s disease (PD)and Parkisonism plus syndrome. Methods The substantia nigra and the striatum of 5 PD cases, 3 progressive supralnuclear palsy (PSP) cases and 3 multiple system atrophy (MSA) cases, and 5 normal aging control cases were examined by routine neuropathological methods and Gallyas-Braak staining and tau, ubiquitin and ?-synuclein immunohistochemistry. Pigmented neurons in the substantia nigra of PD, PSP, MSA cases and normal aging control cases were counted. The neuronal and glial cytoplasmic inclusions in the nigrostriatal tissues were observed. Components of the abnormal proteins were identified. Results Nerve cells in the substantia nigra of PD,PSP and MSA groups showed severe loss in number,especially the ventrolateral zone and the ventromedial zone. Compared with those in the normal aging control group,numbers of nerve cells in the ventrolateral zone of PD, PSP and MSA groups decreased to 37.5%, 24.2%, 33.8% in the right side, and 48.0%, 25.8%, 33.9% in the left side respectively. There were ?-synuclein and ubiquitin-positive Lewy bodies in the substantia nigra of PD. A lot of tau-positive, argyrophilic globous neurofibrinary tangles, tuft-shaped astrocytes and coiled bodies in the substantia nigra and the striatum of PSP were observed.Severe loss of neurons and gliosis in the caudate nucleus and putamen of MSA were found. In addition, ?-synuclein and ubiquitin-positive glial cytoplasmic inclusions were found in the substantia nigra and striatal region of MSA. Conclusions Lewy bodies in PD and glial cytoplasmic inclusions in MSA are related to abnormal depositions of ?-synuclein and ubiquitin.Neuronal and glial cytoplasmic inclusions in PSP are related to abnormal aggregation of tau.

Objective To investigate the effect of anisodamine on calbindin-D28K(CaBP) expression in the ethanol-induced brain damage in rat cerebellum.Methods2 months aged male Sprague-Dawley rats were injected intraperitoeally with ethanol,normal saline,saline+anisodamine and ethanol+anisodamine respectively for 8 d.They were evaluated with Morris water maze.The counts,average area and density of CaBP positive neurons in cerebellum were measured with immunohistochemical technique and image analytical system.Results The latency of Morris water maze was significantly longer in the ethanol group than in the others(P<0.05),while the distance was significantly longer in the ethanol group than in the saline group and saline+anisodamine group(P<0.05).There is not significant difference between ethanol group and ethanol+anisodamine group(P>0.05),but is seemed some longer.The counts,average area and density of CaBP positive Purkinje cell were all significantly less in ethanol group than in the others(P<0.05).There Pwas not significant difference among ethanol+anisodamine group,saline group and saline+anisodamine group(P>0.05) in the counts,but the average areas and density in ethanol+anisodamine group were less than those in saline group and saline+anisodomine group(P<0.05).Conclusion The ethanol can reduce the CaBP expression in the Purkinje cells of the rats cerebellum.Anisodamine can protect the rats cerebellum from it.

BACKGROUND: Synaptic density, a key index of structure and function of brain tissues, is related to cognitive function. Synaptic loss occurs during human brain aging and in Alzheimer disease (AD), inducing the changes of synaptic density.OBJECTIVE: To observe quantitative synaptic alterations in human brain and changes of synaptic density in different parts during normal aging so as to compare them with those of AD patients.DESIGN: Sampling survey.SETTING: Senile Neurological Department of General Hospital of Chinese PLA.PARTICIPANTS: Pathological data were selected from General Hospital of Chinese PLA from June 1996 to December 2002. Inclusion criteria: had no major nervous system diseases and neuropathological changes. Brain tissues of 28 corpses in normal aging group, 23 males and 5 females aged 23-100 years with an average of (65±22.8) years, were obtained at autopsy.All corpses were divided into three groups according to their age, namely,adult group (23-55 years old, n=9), senile group (64-72 years old, n=7),and ＞75 group (76-100 years old, n=12). Cerebral hippocampal samples of other six corpses diagnosed with AD were selected from clinic. The corpses included 5 men and 1 woman aged 76-94 years with an average of (83±7.7) years.METHODS: Response intensity of synaptophysin immunochemistry remained stable after 4-8 hours of death, so brains were obtained at autopsy after 8-72 hours of death and fixed with 4% formalin for at least 6 weeks.In normal aging group, tissues were taken from left superior frontal gyrus,striatal area of left occipital lobe, left putamen (striatum section, including head of caudate nucleus), and left hippocampus (from lateral geniculate body section to medial occipitotemporal gyrus). In AD cases, tissues were taken from left hippocampus of 4 corpses and right hippocampus of other 2. All sections were stained with hematoxylin eosin (HE), toluidine blue and synaptophysin immunostaining (rabbit anti-human synaptophysin polyclonal antibody from Beijing Zhongshan Biotechnology Co., Ltd.). Morphology and distribution of positive objects in synapse immunologic reaction were observed under the light microscope. Relation between absorbance in each region and age was determined with Pearson's coefficient. Differences among groups were analyzed with nonparametric test, and the differences in hippocampal CA3 area between ＞ 75 group and AD group were analyzed with the same test.MAIN OUTCOME MEASURES:① Absorbency of synaptophysin at various sites of normal aging group and correlation with age; ② absorbance value in CA3 area between AD patients and advanced aged normal subjects (＞75 years) was compared.RESULTS:All the 34 cerebral samples entered the final analysis.①Synaptophysin-positive granules of various size were scattered through neocortex, putamen and hippocampus, neuronal somata, neuroglia, vessels and white matter. Density was particularly strong over layers Ⅱ and Ⅲ in frontal lobe, and layer ⅣV in occipital lobe. ② Synaptophysin density was negatively correlated with age, which was -0.688 in frontal lobe, -0.592 in occipital lobe, -0.458 in putamen and -0.619 in hippocampal CA2 area,respectively (P = 0.000, 0.001, 0.014, and 0.000). ③ Significant difference in synaptic density in CA3 area was found between AD patients (0.031 3±0.003 0)and normal subjects over the age of 75 (0.040 7±0.005 3) (Z=-2.997, P=0.001)in nonparametric test.CONCLUSION:① Synaptic density was found to decrease in frontal lobe, occipital lobe, CA3 area of hippocampus and putamen with age; the changes had significant correlation with age.② Synaptic density of AD patients was lower than that of normal subjects, and their cognitive hypofunction was related to synaptic loss. ③ All tissues were obtained after 8-72 hours of death and fixed over 6 weeks, which to the greatest extent reduced the effects of tissue autolysis and formalin fixation on the results.

Objective To construct and identify lentiviral vector pGC-FU-CXCR4 gene. Methods CXCR4 gene amplification was used by real-time polymerase chain reaction. The target gene fragments with the digested plasmids were exchange. Then the lentiviral vector pGC-FU-CXCR4 was constructed successfully. Use the constructed lentiviral vector to infect the competent escherichia coli cells. Polymerase chain reaction analysis was used to identify the cultural clones and DNA sequencing and comparative analysis were used to positive fragments. The successfully constructed plasmids had the same sequence with the target gene. Results Polymerase chain reaction tests showed that am-plified target genes were inserted in pGC-FU vectors. The electrophoresis results,digestion showed that the reconstructed plasmid was consist-ent with the theoretical fragment and the sequence result of the positive fragments were exactly the same with the target gene. Conclusion Lentiviral vectors of CXCR4 gene over-expression were successfully constructed.

Objective To investigate the change of genomic DNA of liver and spleen tissue for different age of the elderly,and provide the experimental data for aging-related research. Methods 35 livers and 33 spleens of autopsied samples preserved in refrigerator at-80 ℃ were divided into 3 groups according to age:age 65y to 79y,age 80y to 89y,age≥90y. The content of DNA in liver and spleen was determined by ultraviolet absorbent method. Results Compaired with age 80y to 89y (0. 310 ± 0. 286)mg/mL,the content of DNA in liver was significant higher at age 65y to 79y (1.464 ±0.488)mg/mL and age ≥90y(1.147 ±0.333)mg/mL(P<0.05);Compared with age 80y to 89y(0. 938 ± 0. 589)mg/mL,the content of DNA in spleen was significant higher at age 65y to 79y(1. 723 ± 0. 726)mg/mL and age≥90y(1. 688 ± 0. 963)mg/mL(P<0. 05). The content of DNA was significant lower in liver (0. 856 ± 0. 658)mg/mL than that in spleen (1. 414 ± 0. 852)mg/mL. Conclusion The content of DNA in human liver and spleen tissue may be decrease along with aging. The content of DNA in the group at age≥90y may be increase. There were some differences between different viscera tissue in content of DNA.

Objective To investigate the effect of Shuyuningxin decoction on the expression of Bcl-2 and Bax in hippocampus of chronicstress depression rats. Methods 60 SD rats were randomly divided into control group, model group, fluoxetine group, high dose ofShuyuningxin decoction group, middle dose of Shuyuningxin decoction group and low dose of Shuyuningxin decoction group. All thegroups were given the chronic unpredicted mild stress for 21 d to induce depression models except the control group. Then they were administratedtheir drugs for 21 d. The level of Bcl-2 and Bax in hippocampus were investigated with immunohistochemistry technique. ResultsCompared with the control group, the level of Bax increased and Bcl-2 decreased in the model group (P<0.05). Compared with modelgroup, the level of Bax decreased and Bcl-2 increased in the middle dose of Shuyuningxin decoction group, high dose of Shuyuningxin decoctiongroup and the fluoxetine group (P<0.05). Conclusion The anti-depression effect of Shuyuningxin decoction may be associated withthe expression of Bax and Bcl-2.