OBJECTIVETo investigate the influence of APP(SWE) on the expression of neuronal acetylcholine receptors (nAChRs) and its relationship with Alzheimer's disease (AD).

METHODSAPP(SWE), carried the Swedish family AD double mutants, were transfected into SH-SY5Y cells and primary cultured neurons from rat brains to build a cellular model of AD. The mRNA levels of APP and nAChRs, and the protein levels of total APP, αAPPs and nAChRs in the cultured cells were measured using real-time PCR and Western blot, respectively. The numbers of α3 nAChR were determined by receptor-[³H]epibatidine binding assay.

RESULTSIncreased expressions of Swedish 670/671 APP at mRNA and protein levels, and down-regulation of αAPPs were observed in both of the cultured neuronal cells transfected with APP(SWE). A significant increase of α7 nAChR expression at protein and mRNA levels was detected in the APP(SWE) transfected SH-SY5Y cells. On the other hand, after transfection with APP(SWE), the expressions of α3 nAChR at protein and mRNA levels in SH-SY5Y cells, and α4 nAChR at mRNA level in primary cultured neurons were inhibited. In addition, the numbers of receptor binding sites were deceased in SH-SY5Y cells overexpressing with APP(SWE).

CONCLUSIONOverexpression of APP(SWE) can decrease αAPPs and modify nAChRs by increasing expression of α7 nAChR and decreasing α3 and α4 nAChRs, which might play an important role in the pathogenesis of AD.

Alzheimer Disease ; genetics ; Amyloid Precursor Protein Secretases ; secretion ; Amyloid beta-Protein Precursor ; genetics ; metabolism ; physiology ; Animals ; Brain Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cells, Cultured ; Cerebral Cortex ; cytology ; metabolism ; Down-Regulation ; Humans ; Neuroblastoma ; metabolism ; pathology ; Neurons ; cytology ; metabolism ; Plasmids ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Nicotinic ; genetics ; metabolism ; Transfection ; alpha7 Nicotinic Acetylcholine Receptor

OBJECTIVETo evaluate the therapeutic effect of individualized defibrase therapy according to the level of plasma fibrinogen (FIB) in patients with acute cerebral infarction (ACI).

METHODSSixty patients with ACI (within 72 h after onset) were randomly divided into defibrase group (n=30) and control group (n=30). The patients in defibrase group received intravenous defibrase infusion at different first doses (15, 10, and 5 U) according to plasma FIB level (>4 g/L, 2-4 g/L, and 1.3-2 g/L) before treatment. Plasma FIB was measured every 12 h after the first dose of defibrase, and when plasma FIB was over 1.3 g/L, intravenous infusion of 5 U defibrase was given to maintain plasma FIB within the range of 0.70-1.13 g/L over a period of 7 days. The plasma prothrombin time (PT), activated partial thromboplastin time (APTT) and FIB before and after the 7-day treatment were measured, and the scores of Chinese stroke scale (CSS) after 14 days of treatment and Activity of Daily Living (ADL) after 3 months were recorded.

RESULTSAfter 7 days of treatment, plasma PT and APTT were significantly prolonged lengthened and plasma FIB was lowered in defibrase group. The scores of CSS improved in defibrase group after 14 days of treatment, showing significant difference from those of the control group. The clinical effective rate was 80% in defibrase group, significantly higher than that in the control group (50%). The scores of ADL after 3 months were similar between the 2 groups, but the percentage of independent living and mild dependency was significantly higher in defibrase group (93.3% vs 70.0%). No intracerebral and extracerebral hemorrhage occurred in defibrase group the during treatment, no did death occur after 3 months of treatment.

CONCLUSIONDefibrase therapy based on plasma FIB level can rapidly and effectively lower plasma FIB, reduce neurological impairment and improve the quality of life in patients with ACI.

Acute Disease ; Adult ; Aged ; Aged, 80 and over ; Batroxobin ; therapeutic use ; Cerebral Infarction ; drug therapy ; Female ; Fibrinogen ; metabolism ; Fibrinolytic Agents ; therapeutic use ; Humans ; Male ; Middle Aged

OBJECTIVESTo investigate the neuroprotective function of alpha 3 nicotinic acetylcholine receptor (nAChR) by inhibiting the gene expression in human neuroblastoma (SH-SY5Y) cells using small interference RNA (siRNA).

METHODSThe siRNA coding oligonucleotide sequences targeting alpha 3 nAChR were designed and synthesized. The annealed product was cloned into pSilencer 3.1-H1 neo vector. The recombinant alpha 3 nAChR pSilencer 3.1-H1 neo vector was transfected into the SH-SY5Y cells. The stable clones were screened by G418 medium, and the levels of alpha 3 nAChR mRNA and protein were monitored by using real-time PCR and Western blotting, respectively. After the SH-SY5Y cells with siRNA treatment were exposed to 1 micromol/L Abeta(1-42), MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide], SOD, GSH-px and the lipid peroxidation were measured by spectrophotometry.

RESULTSCompared with the controls, the expression levels of mRNA and protein in the stable SH-SY5Y clone cells transfected with the recombinant alpha 3 nAChR pSilencer 3.1-H1 neo vector were decreased with inhibitory efficiency of 98% and 66%, respectively, the MTT reduction decreased; the product of lipid peroxidation was increased and the activities of SOD and GSH-px were decreased. Biologically, the gene expression inhibition of alpha 3 nAChR enhanced the toxicity induced by Abeta in SH-SY5Y cells.

CONCLUSIONSThe expression inhibition of alpha 3 nAChR as a result of recombinant alpha 3 nAChR siRNA can induce oxidative stress and improve the toxicity of Abeta on SH-SY5Y cells, indicating that alpha 3 nAChR may play a significant neuroprotective role in the pathogenesis of Alzheimer disease.

Amyloid beta-Peptides ; pharmacology ; Cell Line, Tumor ; Cell Membrane ; drug effects ; Gene Expression Regulation ; drug effects ; Humans ; Neuroblastoma ; pathology ; Oxidation-Reduction ; drug effects ; Peptide Fragments ; pharmacology ; RNA Interference ; immunology ; RNA, Small Interfering ; pharmacology ; Receptors, Nicotinic ; drug effects ; genetics ; metabolism ; Superoxide Dismutase ; antagonists & inhibitors ; genetics ; metabolism

OBJECTIVETo study the effects of beta-amyloid peptide (Abeta) on cell membrane lipids and cholinergic receptors of human neuroblastoma cells.

METHODSHuman SH-SY5Y neuroblastoma cells were treated with different concentrations of Abeta(1-42) with and without pretreatment of vitamin E. MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide] reduction, lipid peroxidation, protein oxidation and phospholipids were measured by spectrophotometry. Levels of cholesterol and unbiquinone were determined by high-performance liquid chromatography (HPLC). The numbers of cholinergic receptor binding sites were determined by receptor binding assay and the protein levels of nicotinic receptor alpha3 and alpha7 subunits were studied by Western blotting.

RESULTSSH-SY5Y cells showed decreased reduction rates of MMT and phospholipids, and increased lipid peroxidation and protein oxidation after exposure to Abeta (0.1 micromol/L) as compared to the control. The number of cholinergic receptor binding sites, the protein level of nicotinic receptor alpha3 and alpha7 subunits and the content of ubiquinone decreased in cells treated with high dose of Abeta (1 micromol/L). Although the level of cholesterol was not changed in any way, vitamin E partially prevented the neurotoxic effects of Abeta.

CONCLUSIONbeta-amyloid peptide reduces the level of cell membrane lipids and cholinergic receptors in human SH-SY5Y neuroblastoma cells, likely through the induction of an enhanced oxidative stress.

Amyloid beta-Peptides ; administration & dosage ; metabolism ; toxicity ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Cholesterol ; metabolism ; Dose-Response Relationship, Drug ; Humans ; Lipid Peroxidation ; drug effects ; Malondialdehyde ; metabolism ; Membrane Lipids ; metabolism ; Neuroblastoma ; metabolism ; pathology ; Oxidative Stress ; drug effects ; Peptide Fragments ; administration & dosage ; metabolism ; toxicity ; Phospholipids ; metabolism ; Receptors, Nicotinic ; metabolism ; Ubiquinone ; metabolism ; Vitamin E ; metabolism ; pharmacology

AIMTo investigate the relation between Glu-R and the protective effect of hypothermia on oxygen and glucose deprivation (OGD) injury in hippocampal slices of rat.

METHODS(1) We had established OGD injury model in rat hippocampal slices. The changes of orthodromic population spike(OPS) during OGD and after administration of hypothermia (32 degrees C, 25 degrees C) were observed. (2) We had established Glu excitatory toxicity injury model in rat hippocampal slices. The changes of OPS after exposure to Glu and the effect of hypothermia (32 degrees C, 25 degrees C) against the Glu excitatory toxicity injury were observed. The non-NMDA receptor-mediated excitatory postsynaptic potentials (EPSP) in the CA1 area were recorded via adding the GABA-R specific agonists bicuculline (BMI) and NMDAR agonists D-(-)-2-Amino-5-phosphonopentanoic Acid (AP5) in normal artificial cerebrospinal fluid (nACSF), the NMDA receptor-mediated EPSP were recorded via adding the BMI and non-NMDA-R agonists 6,7-Dinitroquinoxaline-2, 3(1H,4H)-dione(CNQX) in nACSF. The variety of the changes of OPS during OGD14min in nACSF groups and added BMI compounded AP5 or BMI compounded CNQX ACSF groups were observed after administration of 25 degrees C hypothermia 28 min. (3) The changes of ultrastructure of CA1 area after OGD 1 h and the effect of hypothermia (25 degrees C) on it were observed.

RESULTS(1) OPS reduced and abolished quickly during OGD14min, and the recovery amplitude of OPS was very low after reoxygenation/glucose 1 h. While the time of OPS abolishing significantly elongated and the recovery of OPS was higher in hypothermia (32 degrees, 25 degrees C) groups. The effect in groups 25 degrees C was more significant than those in groups 32 degrees C. (2) In control groups, Glu (2 mmol/L, 14 min) decreased the amplitude of OPS, after the end of Glu exposure the recovery amplitude of OPS was very low. After administration of hypothermia (32 degrees C, 25 degrees C), the recovery amplitude and rate of OPS were significantly higher than those in the control groups, while the antagonism on Glu excitatory toxicity injury in H 25 degrees C was more significant than those in H 32 degrees C. The changes of OPS during OGD 14 min were no distinct difference in nACSF groups and added BMI (50 micromol/L) compounded AP5(20 micromol/L) or BMI (50 micromol/L) compounded CNQX (100 micromol/L) ACSF groups. The protection of hypothermia (25 degrees C) could not be cancelled by added AP5 compounded BMI or BMI compounded CNQX in nACSF. (3) After OGD (14 min) 1 h, the nuclear membrane of pyramidal cells in CA1 area was irregular, nucleus were homogenized, the organelle in the cytoplasm was degenerate, even more to necrosis or loss, mitochondrion swelled, ridge was vacuoles. In H 25 degrees C the nuclear membrane was regular, mitochondrion swelled only lightly. Small chromatin gathered to edge.

CONCLUSIONHypothermia shows the protective effects of against OGD injury in hippocampal slices. The mechanism is related to the antagonism of Glu excitor toxicity and maintenance the ATP level in cells, and the antagonism perhaps is mediated by NMDA-R and non-NMDA-R.

Animals ; Cell Hypoxia ; Glucose ; metabolism ; Glutamic Acid ; toxicity ; Hippocampus ; metabolism ; Hypothermia ; Membrane Potentials ; N-Methylaspartate ; metabolism ; Neurons ; metabolism ; Organ Culture Techniques ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the association between interleukin-10 (IL-10) gene promoter microsatellite polymorphisms and the susceptibility to hepatitis B virus infection in Han,Yi and Yao ethnicities in GuiZhou province.Methods 500 volunteers were selected from Guizhou province.Ailelic frequency of IL-10.G and IL-10.R loci was identified by short tandom repeat polymerase chain reaction.The relativity between allelic frequency and HBV infection was analyzed.Results Genotype data from H-W analysis on all the IL-10 polymorphisms indicated that it was a random distribution.Very high HBV infection rates were found in the native ethnic minorities of Guizhou province.The overall HBV infection rate among the total population was 67.00％,with the HBV infection rates of Yi nationality in Weining,Yi nationality in Qianxi,Yao nationality in Libo and Han nationality in Libo as 51.85％,42.86％,79.52％ and 84.30％,respe~vely.The polymorphisms distribution of IL- 10.G and IL- 10.R were statistically different among the ethnic groups (P＜ 0.05 ).The polymorphisms distribution of IL-10.R had no significant difference between HBV infection group and non-infection group,as well as among HBV natural removal group and non-infected group in all the ethnic groups.The frequency of IL-10.G 459 bp (19CA) was significantly higher in non-infection group than in the infected group (P＜ 0.05 ).The frequency of IL-10.G 471 bp (25CA) was significantly higher in the non-infection group than in the HBV natural removal group(P＜0.05).The polymorphisms distribution of IL-10.G did not show significant difference between the HBV infection group and the HBV natural removal group in all the ethnic groups.We did not find any differences in allelic and genotypic frequencies of IL-10.G between infection group and non-infection group in Yi nationality in Weining,and Yao nationality in Libo (P＞0.05),as well as HBV natural removal group and non-infected group (P＞0.05).Conclusion The polymorphisms distribution of IL-10.R and IL-10.G did not show significant difference in Yi,Yao and Han ethnics population living in Guizhou province.IL-10.G seemed to influence the susceptibility of HBV infection in Han,Yao and Yi ethnics population of Guizhou province.

Objective To carry on a survey on blood routine examination of coal-burning endemic fluorosis population in Bijie City,Guizhou Province in order to study their health status and problems.Methods Blood routine examination was performed in the residents in coal-fired pollution endemic fluorosis-endemic area, including the residents of the Changchun Village of Changcun Town(intervention group)whose stoves had been improved and of Shiba Village Yachi Town not improved in Bijie City,Guizhou Province.The indicators were including leukocyte(WBC),red blood cell(RBC),hemoglobin(Hb),hematocrit(HCT),tlle average hematocrit red blood cell volume(MCV),mean corpuscular hemoglobin(MCH),mean corpuscular hemoglobin concentration (MCHC),red blood cell distribution width-CV(RDW-CV),platelets(PLT).Results RBC,Hb,HCT,MCHC, PLT were(4.95±1.18)×1012/L,(138.46±15.90)g/L,(50.19±11.48)%,(284.90±48.73)g/L,(334.92± 119.34)×109/L for the male in the intervened group,and they were(4.02±0.47)x 1012/L,(131.00±15.90)g/L, (40.90±7.60)%,(323.14±41.95)g/L,(280.79±100.34)× 109/L in non-intervention group,respectively. Inter-group comparison,the difference was statistically significant (U = 7.72,3.50,7.12,6.28,3.66,P ＜ 0.01). RBC, HCT,MCV,MCH,MCHC,RDW-CV,PLT were respectively(4.75±1.20)×1012/L,(46.91±11.20)%,(99.30± 6.88)fl,(28.10±8.66)pg,(275.61±54.49)g/L,(16.95±1.63)%,(351.23±150.37)×109/L for the female in the intervened group,and were (3.85±0.65)×1012/L,(38.80±6.60)%,(100.80±7.00)fl,(33.10±5.40)pg, (327.14±44.52 ) g/L,(16.60±1.58) %,(279.40±98.07)×109/L in the group un-intervened. Inter-group comparison found that there was a significant difference(U = 8.92,10.72,2.04,6.61,9.82,2.06,5.39,P ＜ 0.001 or 0.05) and the abnormal rate of RBC and Hb in non-intervention group[ 32.62% (92/282),16.67%(47/282)] was higher than that in the intervention group[9.73%(29/298) ,6.71%(20/298),x2 = 45.992,14.054,P ＜ 0.01 ) ]. Conclusion Experiment group has better results of blood routine test compared to non-intervention group,especially of anemia.

OBJECTIVETo observe the effect of acupuncture combined with speech therapy for dysarthria after stroke or cerebral trauma.

METHODSSixty-one cases were randomly divided into two groups. The observation group (30 cases) was treated with speech therapy and acupuncture at Lianquan (CV 23), Jinjin (EX-HN 12), Yuye (EX-HN 13), Fengchi (GB 20), Yifeng (TE 17) and Wangu (GB 12) as major acupoints, while the control group (31 cases) was treated with speech therapy only. The changes of speech and acoustics indices were evaluated after 9 weeks treatment.

RESULTSThe word articulation and correct rate of text of patients in two groups were both obviously improved after treatment (both P < 0.01). The total effective rate of 96.7% (29/30) in observation group was superior to that of 67.7% (21/31) in control group (P < 0.01). The maximum phonation time (MPT) of patients tested by aeromechanics analyzer were obviously prolonged in observation group (12 cases) and control group (11 cases) (both P < 0.01), and the improvement in observation group was more obvious (P < 0.01).

CONCLUSIONAcupuncture combined with speech therapy can improve the effect on language and acoustics level for dysarthria.

Acupuncture Therapy ; Adult ; Aged ; Dysarthria ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Speech Acoustics ; Speech Therapy ; Young Adult

OBJECTIVETo analyze the epidemiologic characteristics of severe acute respiratory syndrome (SARS) and to evaluate the effectiveness on its major control measures in Tianjn.

METHODSAdopting two case reports 1 and 2, designed by the Tianjin Centers for Disease Control and Prevention to develop a unified case-tracing table including the map of the distribution of close contacts to SARS patients. With those methods, investigation on patients and their close contacts at hospital wards, families, communities and institutions of the patients were carried out.

RESULTSFrom April 13 through May 8, 2003, there were 175 SARS cases including imported ones, were identified with an incidence rate of 1.9 cases per 100,000. Among them, 14 died with a fatality of 8.0%. The whole process of epidemic in Tianjin was less than one month with the following features: (1) 93.7% of the total SARS cases in Tianjin were directly or indirectly transmitted by a super-spreader. (2) 68.6% of the total SARS patients were concentrated in 3 hospitals A, B and C which was menifastated in 'clustering'. Through study on the rest of the SARS patients, results showed that 16.8% of them were transmitted through family close contact and 2.3% due to contact to colleagues. However, 12.6% of the patients were not able to show evidence that they had any contact to a diagnosed SARS patient. At the early stage of the epidemic, a number of medical practitioners were infected, taking up 38.2% of the total SARS cases. Among the total number of 1 975 medical workers who participated in the SARS medical cares and treatments, 3.4% of them got infected. During the outbreak, all index cases and chains of transmission seemed to be clear, with only 3 patients not able to be traced for the source of infection, taking up 2% of the total SARS patients in Tianjin. Among the 10 index cases, only the super spreader and another one index case transmitted the virus to their contacts but the rest of index cases did not cause any secondary infection.

CONCLUSIONThough SARS is clinically severe and can be spreaded quickly, the epidemic can be under control within a short period of time if chains of SARS transmission are broken down and effective measures as isolation and quarantine against patients as well as underscoring awareness among the publics in a scientific way, being carried out.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Contact Tracing ; Cross Infection ; transmission ; Disease Outbreaks ; Family Health ; Female ; Humans ; Incidence ; Infectious Disease Transmission, Patient-to-Professional ; Male ; Middle Aged ; Severe Acute Respiratory Syndrome ; epidemiology ; mortality ; transmission ; Surveys and Questionnaires

OBJECTIVETo establish homozygous transgenic mouse strain expressing human tau isoform with P301L mutation.

METHODSFive transgenic mice expressing human tau isoform with P301L mutation were obtained by microinjection into male nuclei. Homozygote and hemizygote were identified by PCR and real-time fluorescent quantitative PCR.

RESULTSNinety five homozygous transgenic mice were selected, and the results indicated that homozygous transgenic mice were superior to hemizygote in simulating the changes of biological characteristics.

CONCLUSIONExogenous gene tau is able to stably transmit to next generation and the combination of SYBR Green real-time fluorescent quantitative PCR with the traditional mating is a fast, reliable and economical way to screen homozygous and hemizygous transgenic mice.

Animals ; Female ; Homozygote ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microinjections ; Mutation ; tau Proteins ; genetics