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Objective:To observe the effect of tetrahydroxy stilbene glycoside (TSG) on the expression of glycogen synthase kinase 3<italic>β </italic>(GSK3<italic>β</italic>), cyclic adenosine monophosphate-dependent protein kinase (PKA) and Serine/threonine phosphatase 2A(PP2A) in the brain of amyloid precursor protein/presenilin-1/Tau (APP/PS1/Tau) triple-transgenic mice dementia model. Method:A total of forty-five 8-month-old APP/PS1/Tau transgenic mice were randomly divided into model group, positive control group (Huperzine-A, 0.15 mg·kg<sup>-1</sup>), low, medium and high dose TSG groups (TSG, 0.033,0.1,0.3 g·kg<sup>-1</sup>), with 9 mice in each group, and another nine C5B7L/6J mice of the same age were selected as normal control group. After 60 days of intragastric administration, the general structure of hippocampal neurons was observed by hematoxylin-eosin (HE) staining, immunohistochemical (IHC) was used to detect the expression of PKA protein in the brain of mice in each group, the mRNA expression levels of GSK3<italic>β</italic>, PKA and PP2A were detected by real time quantitative reverse transcription polymerase chain reaction (Real-time PCR), and protein expression levels of GSK3<italic>β</italic> and PP2A were detected by Western blot. Result:Compared with the normal control group, the apoptosis level of neurons in the model group was significantly increased, the protein and mRNA expression levels of GSK3<italic>β</italic> and PKA were significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01), and the protein and mRNA expression levels of PP2A were significantly decreased (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with the model group, the apoptosis level of neurons in each treatment group was significantly down-regulated, the protein and mRNA expression levels of GSK3<italic>β</italic> and PKA were significantly down-regulated (<italic>P</italic><0.05, <italic>P</italic><0.01), and the protein and mRNA expression levels of PP2A were significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01). Conclusion:The mechanism of TSG in the treatment of Alzheimer's disease (AD) may be related to lowering the transcription and expression of GSK3<italic>β</italic> and PKA, increasing the transcription and expression of PP2A.
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Objective To investigate the mutations in methyl-CpG-binding protein 2 gene (MECP2 gene) from typical sporadic Rett syndrome patients,explore the correlations between their genotype and phenotype,assist in genetic counseling.Methods Genomic DNA was extracted from peripheral blood leukocytes from 2 patients and their parents using standard protocols.Polymerase chain reaction and direct sequencing were performed using specific primers from 4 exons in MECP2 gene.Results No mutations were found in exon 1,2,3.Two different heterozygous missense mutations in exon 4 within MECP2 gene were identified from 2 patients.Their nuclear acid changes were:c.C473T and c.C397T,leading to amino acid change accordingly:p.T158M and p.R133C.There were no same mutations from their parents.Phenotype of patient with c.C397T was milder than patient with c.C473T.Conclusions Most of typical Rett syndrome patients had mutations in MECP2 gene.Gene test should be performed.Their biological parents should be detected accordingly if the patient had positive found to support genetic counseling.
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Objective By sequenceing the Cj1136,Cj1138 and Cj1139 gene of Campylobacter jejuni(C. Jejuni) strains associated with Guillain-Barre Syndrome(GBS),features of Cj1136,Cj1138 and Cj1139 gene were studied.Results were compared with the C.jejuni strain NCTC11168, to find the mutations in sequence of C.jejuni which inducing GBS and their polygenetic relationship was analyzed.Methotis Three GBS-associated C.jejuni strains were isolated from stools of GBS patients from Hebei province who had been diagnosed as clinical AMAN pattern and electrophysiological tests were performed.After distilling and sequencing Cj1136,Cj1138 and Cj1139 genes,results were spliced and assembled into a complete sequence by the terminals overlapped with each other.Sequences of Cj1136,Cj1138 and Cj1139 genes were compared with NCTC11168,to find the mutations and gene feature.Results The Cj1136,Cj1138 and Cj1139 gene of the three GBS-associated C.jejuni strains were composed by 1173 base pairs,1170 base pairs,912 base pairs respectively. The alignment with the related sequence of NCTC11168 showed that there were two same mutations in the Cj1138 gene of the three C.jejuni stains.Data from phylogenetic analysis demonstrated that the three C.jejuni strains were genetically closed to NCTC11168,with the biggest phylogenetic distance between the three of them as 2.1%.Conclusion When compared with NCTC11168 the Cj1138 gene of the three GBS-associated C.jejuni strains had the same mutations which might be related to the development of GBS.Relation between the variation and GBS-pathogenesis remained to be confirmed.The mutations found in the three C.jejuni strains established the foundation for exploring the biological characteristics of GBS-associated C.jejuni strains and demonstrated that the GBS-associated C.jejuni strains of Hebei province having its regional features.
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<p><b>OBJECTIVE</b>To investigate the regulatory effects of nerve growth factor (NGF) on basal and capsaicin-induced release of neuropeptide substance P (SP) in primary cultured embryonic rat dorsal root ganglion (DRG) neurons.</p><p><b>METHODS</b>DRGs were dissected from 15-day-old embryonic Wistar rats. DRG neurons were dissociated and cultured, and then exposed to different concentrations of NGF (10 ng/mL, 30 ng/mL, or 100 ng/mL) for 72 h. The neurons cultured in media without NGF served as control. RT-PCR were used for detecting the mRNAs of SP and vanilloid receptor 1 (VR1) in the DRG neurons. The SP basal and capsaicin (100 nmol/L)-induced release in the culture were measured by radioimmunoassay (RIA).</p><p><b>RESULTS</b>SP mRNA and VR1 mRNA expression increased in primary cultured DRG neurons in a dose-dependent manner of NGF. Both basal release and capsaicin-evoked release of SP increased in NGF-treated DRG neurons compared with in control group. The capsaicin-evoked release of SP also increased in a dose-dependent manner of NGF.</p><p><b>CONCLUSION</b>NGF may promote both basal release and capsaicin-evoked release of SP. NGF might increase the sensitivity of nociceptors by increasing the SP mRNA or VR1 mRNA.</p>
Subject(s)
Animals , Rats , Analgesics, Non-Narcotic , Pharmacology , Capsaicin , Pharmacology , Cells, Cultured , Dose-Response Relationship, Drug , Embryo, Mammalian , Ganglia, Spinal , Cell Biology , Gene Expression Regulation , Nerve Growth Factor , Pharmacology , Neurons , RNA, Messenger , Metabolism , Radioimmunoassay , Methods , Rats, Wistar , Substance P , Genetics , MetabolismABSTRACT
<p><b>AIM</b>SD rats were utilized for the purpose of the exploration of effects of status epilepticus (SE) on their emotional behavior, spatial learning and memory, and explorating its molecular mechanism.</p><p><b>METHODS</b>Forty maturity male SD rats, weighing (200 +/- 20) g were divided randomly and equally into SE group (SG) and normal control group (NG). The SG rats were induced by Pentylenetetrazole (PTZ) and the control animals received a saline (0.9%) solution. The change of emotional behavior in two groups were tested in elevated plus maze. Furthermore, Morris water maze was applied to evaluate the effects by SE on spatial learning and memory in rats. At the same time, N-methyl-D-aspartate (NMDA) receptor NR1 subunit mRNA in the hippocampus was determined by reverse transcription polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>In elevated plus test, SE rats increased the times of visits as well as the time spent on the open arms of the elevated plus maze (P < 0.01). In Morris water maze, the mean escape latency for the SE rats looking for hidden platform in the place navigation test prolonged (P < 0.01). The efficiency of their search strategy was poor (P < 0.05). The swimming time in platform region and the percentage of their swimming time decreased (P < 0.01). The number of times they crossed the platform area decreased (P < 0.01). Meanwhile the expression of NR1 subunit mRNA in hippocampus was lower (P < 0.01).</p><p><b>CONCLUSION</b>The experimental results showed that SE could result in the change of emotional behavior and damage of spatial learning and memory in rats. NR1 might be involved in the patho- and physiological process in causing these behavioral changes.</p>