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Objective:To investigate the effects of cattle encephalon glycoside and ignotin(CEGI)on the expression of glial fibrillary acidic protein(GFAP)and neuronal nuclear antigen(NeuN)in the brain of APP/PS1 model mice of Alzheimer's disease.Methods:A total of 36 5-month-old APP/PS1 dual-transgenic model mice were randomly divided into 3 groups: the model group(normal saline 6.6 ml·kg -1·d -1), CEGI group(CEGI 6.6 ml·kg -1·d -1)and donepezil group(donepezil 2 mg·kg -1·d -1), with 12 in each group.Twelve C57BL/6J mice of the same age were used as the normal control group.All mice were given drugs for 6 weeks consecutively.Brain tissue was collected for immunohistochemical staining to detect the expression of amyloid β-protein(Aβ), GFAP and NeuN, which were then analyzed quantitatively. Results:The results of immunohistochemical staining indicated that levels of Aβ and GFAP were higher and levels of NeuN were lower in the model group than in the normal control group(0.147±0.068% vs.0%, 61.750±22.020 vs.26.000±4.598, 0.021±0.002 vs.0.032±0.003, P<0.05). Levels of Aβ and GFAP were lower and levels of NeuN were higher in the CEGI group and the donepezil group than in the model group(0.058±0.055 % vs.0.057±0.045 %, 38.250±5.418 vs.36.130±5.963, 0.029±0.004 vs.0.027±0.003, P<0.05). There was no significant difference in the expression of Aβ, GFAP and NeuN between the CEGI group and the donepezil group( P>0.05). Conclusions:CEGI has multi-target neuroprotective effects via down-regulating the expression of Aβ and GFAP and up-regulating the expression of NeuN.
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Objective To study the pathological features of argyrophilic grains in normal aging brain, AD, PD and progressive superior nuclear paralysis patients. Methods Brain tissue samples taken from 5 AD, 3 PD, 2 progressive superior nuclear paralysis patients with complete clinico-pathological data and 4 normal aging brain subjects were stained with HE, Luxol fast blue and Gallyas-Braak silver respectively. Aβ, tau, α-synuclein and P62 antibodies were detected by microscopy with immunohistochemical staining. The pathological features of argyrophilic grains were recorded. Results The Gallyas-Braak silver staining showed argyrophilic grain structure in 4 out of the 14 amygdaloid nucleus tissue samples (2 from AD patients, 1 from progressive superior nuclear paralysis patients and 1 from normal aging brain patients) with a positive rate of 28.6%. The immunohistochemical staining showed positive tau and P62 antibodies. Conclusion Argyrophilic grain lesion is not uncommon in aging-related neurodegenerative diseases such as normal aging brain, AD and progressive superior nuclear paralysis and can thus produce its superposition effect on the clinical symptoms of cognitive impairment in AD and progressive superior nuclear paralysis patients.
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Objective@#To search for biomarkers for human familial prion disease.@*Methods@#Two-dimensional differential gel electrophoresis (2D-DIGE) proteomic analysis has been performed in frontal lobe tissues of 3 patients suffering from human familial prion disease (PrP) and 3 age-and sex-matched patients suffering from sudden death due to heart failure without neurological disease.@*Results@#The maps revealed 14 polypeptide chains differentially modulated in the PrP samples, among those, 7 could be identified upon digestion and MALDI-TOF/MS analysis, of which 6 appeared to be up-regulated, 1 being down-regulated.@*Conclusions@#We highlight Galectin-1(Gal-1), ryanodine receptor 2 (RyR2), ubiquitin, Rab-interacting lysosomes protein-like protein 1 (RILPL-1) profillin 2 (PFN2), in the differential map. These proteins are related to neurogenesis, the clearance of misfolded proteins, stasis of calium channel, myoclonus and so on. These proteins are potential biomarkers or targets for treatment of prion disease.
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<p><b>OBJECTIVE</b>To investigate histopathology and proteinopathy in the spinal cord of patients with common neurodegenerative diseases.</p><p><b>METHODS</b>Spinal cord tissues from clinically and neuropathologically confirmed neruodegnerative diseases were enrolled in this study, including 3 cases of multiple system strophy, 4 cases of amyotrophic lateral sclerosis, 5 cases of Alzheimer's disease (AD, included 2 cases of AD combined with Parkinson's disease), 2 cases of progressive supranuclear palsy, 1 case of dementia with lewy body and 1 case of corticobasal degeneration from 1955 to 2013 at Chinese People's Liberation Army General Hospital. Four normal control cases were also included. Routine HE and Gallyas-Braak staining, and immunohistochemical stainings for anti-PHF tau (AT8), anti-α-synuclein, anti-TDP-43 and anti-ubiquitin were performed.</p><p><b>RESULTS</b>Examination of the spinal cord in 3 cases with multiple system strophy revealed severe neuron loss in the intermediolateral nucleus of thoracic segment and Onuf's nucleus of the sacral segment, along with moderate neuron loss in the anterior horn of the cervical segment and mild myelin pallor in the anterior funiculus and anterolateral funiculus in the cervical and thoracic segments. Large amount of argentophilic, ubiquitin and synuclein positive oligodendroglial cytoplasmic inclusions were found widely distributed in the anterior horn and the anterior funiculus and anterolateral funiculus of the full spinal cord. Severe neuron loss and several morphological changes with gliosis in the anterior horn and severe loss of myelin in the anterior funiculus and anterolateral funiculus of the full spinal cord were observed in 4 cases of amyotrophic lateral sclerosis, 2 of which were found with Bunina bodies in neurons of the anterior horn. Three amyotrophic lateral sclerosis cases had ubiquitin-positive neuronal inclusions and TDP-43 positive neuronal and glial inclusions in the anterior horn at cervical and lumbar segments. A few argentophilic, tau positive neurofibrillary tangles (NFTs) and neuropil threads in the anterior horn at cervical and lumbar segments were found in 4 AD cases. Examination of spinal cord in 2 cases with Parkinson's disease combined with AD and 1 case with dementia with lewy body revealed severe neuron loss in the intermediolateral nucleus of thoracic segment, and a few synuclein positive lewy bodies and neuritis were also observed. There was mild neuron loss in the anterior horn at cervical and lumbar segments, along with some argentophilic, tau positive globous NFTs and many argentophilic, tau positive neutrophil threads were observed in 2 progressive supranuclear palsy cases and 1 corticobasal degeneration case.</p><p><b>CONCLUSION</b>Each common neurodegenerative diseases of the spinal cord including multiple system strophy, amyotrophic lateral sclerosis and Parkinson's disease has its own specific histopathology and proteinopathy characteristics.</p>
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Humanos , Enfermedad de Alzheimer , Patología , Esclerosis Amiotrófica Lateral , Patología , Proteínas de Unión al ADN , Metabolismo , Inmunohistoquímica , Cuerpos de Inclusión , Patología , Enfermedades Neurodegenerativas , Patología , Ovillos Neurofibrilares , Patología , Neuronas , Patología , Enfermedad de Parkinson , Patología , Médula Espinal , Patología , Ubiquitina , Metabolismo , alfa-Sinucleína , MetabolismoRESUMEN
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.
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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.
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<p><b>OBJECTIVE</b>To recognize relationship of protein related neurodegeneration abnormal aggregation in the aged brains with their cognitive and motor functions.</p><p><b>METHODS</b>Brain tissues from the consecutive autopsy cases of the aged from January 2005 to December 2006 in PLA General Hospital were carried out for immunohistochemical staining with beta amyloid, tau, α-synuclein and ubiquitin antibodies. The consortium to establish a registry for Alzheimer's disease (CERAD) was used to semi-quantitatively analyze Aβ positive core plaques density and Braak staging for tau positive neurofibrillary tangles (NFTs) and α-synuclein positive Lewy bodies. In addition, Aβ positive cerebral amyloid angiopathy (CAA), neuritic plaques and various ubiquitin positive structures were also observed. The relationship of these protein abnormal depositions in the aged brains with cognitive and motor functions were analyzed.</p><p><b>RESULTS</b>In brain tissues of 16 consecutive autopsy cases of the aged from 78 to 95 years, there were 13 cases with Aβ positive core plaques, their density was 2 cases with sparse, 2 cases with moderate and 9 cases with frequent, respectively, according to CREAD.Eight cases with Aβ positive CAA were found, including 6 cases of mild CAA and 2 cases of severe CAA. There were 12 cases with tau positive NFTs, including 6 cases with Braak stageI-II, 4 cases with stage III-IV and 2 cases with stage V-VI. There were 5 cases with frequent Aβ core plaques, meanwhile existing numerous tau/ubiquitin positive neuritic plaques and Braak stage IV-VI of tau positive NFTs, all of them presented cognitive dysfunction. Among 4 other cases with frequent Aβ core plaques, only one case coexisted α-synuclein positive Lewy bodies showed moderate cognitive impairment, remaining 3 cases did not present cognitive dysfunction. There were 4 cases with α-synuclein positive Lewy bodies in the brainstem, and all of these cases presented parkinsonian motor dysfunction. 13 cases with ubiquitin positive structures were found.</p><p><b>CONCLUSIONS</b>Beta amyloid protein positive deposit in the aged brain is an important marker of normal brain aging and cognitive impairment; frequent Aβ core plaques in the neocortex plus Braak IV and above tau positive NFTs are closely related to cognitive dysfunction of Alzheimer's disease; α-synuclein positive Lewy bodies in the brainstem is one of the important pathological markers of parkinsonian motor disorders; ubiquitin deposition involves the development of some characteristic structures of several neurodegenerative diseases.</p>
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Anciano , Humanos , Enfermedad de Alzheimer , Metabolismo , Patología , Péptidos beta-Amiloides , Autopsia , Encéfalo , Patología , Química Encefálica , Angiopatía Amiloide Cerebral , Ovillos Neurofibrilares , Química , Patología , Placa Amiloide , Ubiquitina , alfa-Sinucleína , Proteínas tauRESUMEN
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.
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<p><b>OBJECTIVE</b>To assess the association of vitamin D receptor (VDR) gene Fok I and Bsm I polymorphisms with dyslipidemia in elderly male patients with type 2 diabetes of Han nationality.</p><p><b>METHODS</b>A total of 328 elderly male residents of Han nationality in Beijing, including 237 type 2 diabetic patients and 91 healthy control subjects, were enrolled in this study. The diabetic patients were divided into non-dyslipidemia group (DO group, n=134) and dyslipidemia group (DH group, n=103). All the participants were genotyped for Fok I and Bsm I polymorphisms in VDR gene using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing technology, and the results were compared with their clinical characteristics.</p><p><b>RESULTS</b>For Fok I, the frequency of F allele was significantly higher in the diabetic patients than in the control group (Χ(2)=3.873, P=0.049, OR=1.439, 95% CI: 1.001-2.071). In the dominant model, the frequency of FF genotype was significantly higher in the diabetic group (Χ(2)=5.057, P=0.025, OR=1.756, 95% CI: 1.072-2.875) as well as in DH group (Χ(2)=6.168, P=0.013, OR=2.06, 95% CI: 1.161-3.663) than in the control group. There was no significant differences in the genotype frequency or allele distribution in other paired groups (P>0.05). Compared with Ff + ff genotype, FF genotype was associated with a significantly decreased average diastolic blood pressure (P=0.039) but significantly increased postprandial blood glucose (P=0.035), triglycerides (P=0.049) and uric acid (P=0.031). No significant difference was detected in genotype frequency or allele distribution of Bsm I polymorphisms between the groups (P>0.05); serum creatinine levels were significantly higher in bb genotype than in BB + Bb genotype group (P=0.011).</p><p><b>CONCLUSION</b>VDR gene Fok I polymorphisms may be a risk factor for dyslipidemia in elderly male patients with type 2 diabetes among Chinese Han population, where Bsm I polymorphisms are not associated with diabetic dyslipdiemia.</p>
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Anciano , Humanos , Masculino , Alelos , Glucemia , Presión Sanguínea , Estudios de Casos y Controles , Diabetes Mellitus Tipo 2 , Genética , Dislipidemias , Genética , Etnicidad , Genotipo , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Receptores de Calcitriol , Genética , Factores de Riesgo , Triglicéridos , SangreRESUMEN
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.
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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.
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@#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.
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BACKGROUND: Vitamin D receptor (VDR) gene shows restriction fragment length polymorphism with incision enzyme Bsm Ⅰ ,Apa Ⅰ ,Taq Ⅰ ,which is related to bone mineral density (BMD).However, it is unclear that the relationship between VDR gene (Bsm Ⅰ ) polymorphisms and BMD,osteoporosis.OBJECTIVE: To analyze the distribution regularity of vitamin D receptor gene polymorphism related to BMD in postmenopausal women of Han,Uygur, Kazak and Mongoloid nationality in China. DESIGN: controlled observation.SETTING: Institute of Gerontology,General Hospital of Chinese PLA.PARTICIPANTS: Totally 179 women of Han,who were taking physical examination in General Hospital of Chinese PLA from January 2002 to December 2003, at the average age of (59±3) years,were selected. A total number of 122 women of Uygur with average age of 56.49 years; 63 women of Kazak with average age of (55±3) years; and 112 women of Mongoloid with average age of (57±3) years,who were all taking physical examination in department of geriatrics, Urumqi General Hospital of Lanzhou Military Area Command of Chinese PLA from January 2001 to December 2003.All of them were informed consent.METHODS: VDR genotypes(Bsm Ⅰ ) were defined with polymerase chain reaction-restriction fragment length polymorphism,so as to analyze distribution of Vitamin-D receptor gene (Bsm Ⅰ )polymorphisms of postmenopausal women in Han,Uygur, Kazak,Mongoloid nationality,and compared with the data of USA,Australia,France,Japan,Korea. Enumeration data were compared with Chi-square test.MAIN OUTCOME MEASURES: VDR (Bsm Ⅰ ) polymorphisms in healthy postmenopausal women from Han, Uygur, Kazak, Mongoloid populations in China, which were compared with the data of USA, Australia,France, Japan, Korea.RESULTS: For women of Han, Uygur, Kazak and Mongoloid nationality,the BB genotypes accounted for 0, 4.1%, 6.35% and 4.46%, the bb genotypes accounted for 90.5%, 69.67%, 38.1% and 50% respectively. There was a significant difference between women of Han, Uygur, Kazak, and Mongoloid nationality(P < 0.01). There was insignificant difference in comparation of distribtuion of VDR genotype between Kazak nationality and the west races, but it was significantly different to that in Japan,Korea races.CONCLUSION:VDR genotype polymorphisms is characterized by obvious racial diversify in postmenopausal women of Han,Uygur, Kazak,Mongoloid populations in China;Distribution of VDR gene frequency of Kazak population is similar with the west race ,but is different to Japanese and Korea's race.
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BACKGROUND: It is found reported that polymorphism of Fok 1 restriction endonuclease cut site on exon 2 of 5' end start codon of 5' end start codon (SC), which affected the structure of VDR amino acids,and was relative related to bone mineral density(BMD).OBJECTIVE: To analyze the association between Vitamin D receptor gene (Fok 1) polymorphisms and osteoporosis in the elderly men.DESIGN: case-controlled trialstudy.SETTING: Institute of Gerontology, Chinese PLA General Hospital and Department of Endocrinology,Second Artillery General Hospital of Chinese PLA.PARTICIPANTS: A total of 26 elderly men with osteoporosis at out-patients clinic of Chinese PLA General Hospital and Department of Endocrinology,Second Artillery General Hospital of Chinese PLA from January 2002 to June 2002 were selected involved as osteoporosis case group,with and the average age of was (70±5) years, and BMD in osteoporosis group was 2.0-2.5 SD lower than 2.0-2.5 SD of the peak of BMD. Totally 66 healthy men with average age of (70±5)years were selected as control group during at the same time. All the subjects signed the informed consent,who were Beijing inhabitants of Han nationality, and there was no blood relationship among them.METHODS:VDR-Fok1 genotypes in both groups were detected with by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP),and distributiondistribution of VDR-Fok 1 genotypes were analyzedanalyzed.MAIN OUTCOME MEASURES: distribution Distribution of VDR-Fok1genotypes in both each groups.RESULTS: Totally 66 healthy elderly men and 26 elderly men with osteoporosis entered analysis of results. The frequencies of FF, Ff and ff genotype were found to be 42%, 42% and 15% in control group, and 15%,50%,35% in osteoporosis group, respectively,and there was significantly different between two groups(x2=12.078,P < 0.01).Frequency of allele were significantly different between control group and osteoporosis group (64%,36% vs 40%,60%, x2=8.232,P < 0.01).CONCLUSION: There is a significant difference in the frequency distrinution of VDR gene start codon polymorphism between healthy elderly men and those with osteoporosis.
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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.
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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
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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.