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Objective To study the effect of Tongbiding injection on expression of N-methyl-D-aspartate (NMDA)receptor 1 mRNA in the spinal cord of chronic constriction injury(CCI)rats.Methods Thirty male Spragru-Dawley rats were randomly divided into control group(group A),CCI group(group B)and Tongbiding group(group C),ten rat for eah group.The changes in NR1 mRNA expression in the spinal cord was detected by reverse transcription-polymerase chain reaction(RT-PCR)techniques.Results The expression of NR1 mRNA in the spinal cord was minimal in control group,CCI induced significant increase in the expression of NR1 mRNA in the spinal cord and Tongbiding significantly inhibited the increase in NR1 mRNA expression.There was significant difference between CCI and Tongbiding group(P
ABSTRACT
Objective To study the effect of Tongbiding freezing-dryied powder for injection on the function of WDR neuron of spinal cord in living rats with chronic constriction injury (CCI),and clarify its analgesia mechanism in spinal cord center,and further explore whether it has the drug dependence at the cellular level. Methods Electricity physiology extracellular recording technique was used ,in the CCI living rat's expanded spinal cord waist stage the electricity activity of identical WDR neuron cells was recorded before and after administration of 20 mg?kg-1 Tongbiding. The electric discharge number of C response was observed before and 2,4,8,10 min after administration. The spontaneous electric discharge number. was observed before and 1,2,4,6 min after administration; wind-up phenomenon was also observed.Results The electric discharge number of C response was obtained in 8 WDR neurons,three were significant differences of the mean electric discharge number of C response between 2,4,8 min after administration and before administration (P
ABSTRACT
Objective To construct the eukaryotic expression recombinant PIAS3 plasmid fused with Myc protein and express the fusion protein Myc-PIAS3.Methods The full length PIAS3 fragment of 1 851bp was amplified by PCR and ligated into pMD18-T vector. The full length PIAS3 fragment was subcloned into eukaryotic pCMV-Myc vector between SalⅠ and NotⅠ sites.The recombinant pCMV-Myc-PIAS3 plasmid was trandfected into PC3 cells.The eukaryotic expression Myc-PIAS3 protein was checked by Western blotting with Myc antibody.Results The recombinant plasmid showed right sequence by the full length sequencing.The recombinant plasmid of pCMV-Myc-PIAS3 was identified by enzyme digestion.As expected,by EcoRⅠ digestion,it showed two bands of 4 357bp and 1 318 bp. By XbaⅠdigestion,it showed two bands of 3 291 bp and 2 384 bp.The sequencing result showed a N-terminal Myc of 13 amino acids followed with PIAS3 gene sequence in right reading frame.The pCMV-Myc-PIAS3 plasmid was transfected into prostate cancer PC3 cells.A specific protein expression band at relative molecular mass 68 000 was obtained by using Myc-antibody with Western blotting method.Conclusion The recombinant plasmid of pCMV-Myc-PIAS3 is sucssesefully constructed,and Myc-PIAS3 fusion protein is sucssesefully expressed.