ABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of Snitrosylation of protein disulphide isomerasec in methamphetamine (METH)-induced expression of alpha synuclein (αSN) in mouse hippocampus and striatum neurons.</p><p><b>METHODS</b>Forty C57BL/6 mice were randomized equally into saline control group, METH group, L-NNA (a NOS inhibitor) group and L-NNA plus METH group. All the agents were injected intraperitoneally at an interval of 12 h, and a total of 8 injections were administered; in L-NNA plus METH group, METH was injected 30 min after LNNA in each treatment. Western Blotting was used to detect the expression of nitric oxide synthase (NOS), αSN, PDI and Snitrosylation of protein disulphide isomerase (PDI-SNO) in the hippocampus and striatum of the mice, and nitric oxide (NO) levels were determined using a NO assay kit.</p><p><b>RESULTS</b>In METH group, the levels of NOS, PDISNO, αSN and NO all increased significantly compared with those in the control group (P<0.05). Combined treatment with L-NNA and METH, compared with METH alone, resulted in significantly lowered expression of NOS, NO, PDI-SNO and αSN in the hippocampus and striatum of the mice (all P<0.05). No significant differences were found in NOS, NO, PDI-SNO or αSN expressions among METH+L-NNA, L-NNA and control groups (P>0.05).</p><p><b>CONCLUSION</b>METH induces the activation of NOS and increases NO level to cause the occurrence of PDI-SNO, leading subsequently to increased expression of αSN in mouse striatum and hippocampus. L-NNA, the inhibitor of NOS, can partly relieve nervous system toxicity induced by METH.</p>
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of methamphetamine (METH) exposure on S-nitrosylation of protein disulphide isomerase and the neurotoxicity of METH in PC12 cells.</p><p><b>METHODS</b>PC12 cells were exposed to different concentrations of METH, and the cell viability was assessed using the cell-counting kit-8. PC12 cells exposed to METH in the presence of the NOS inhibitor N-nitro-L-arginine (L-NNA) were examined for cell viability and S-nitrosylation of protein disulphide isomerase using the biotin-switch method, and the changes in cell morphology were examined with HE staining.</p><p><b>RESULTS</b>METH exposure obviously decreased the cell viability and increased S-nitrosylation of protein disulphide isomerase, and the effect of METH was obviously inhibited by L-NNA treatment.</p><p><b>CONCLUSION</b>METH can cause obvious neurotoxicity and promote S-nitrosylation of protein disulphide isomerase in PC12 cells.</p>
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the value of PowerPlex21 System (Promega) and study the genetic polymorphism of its 20 short-tandem repeat (STR) loci in southern Chinese Han population.</p><p><b>METHODS</b>We conducted genotyping experiments using PowerPlex21 System on 20 autosomal STR loci (D3S1358, D1S1656, D6S1043, D13S317, Penta E, D16S539, D18S51, D2S1338, CSF1PO, Penta D, TH01, vWA, D21S11, D7S820, D5S818, TPOX, D8S1179, D12S391, D19S433 and FGA) in 2367 unrelated Chinese Han individuals living in South China. The allele frequencies and parameters commonly used in forensic science were statistically analyzed in these individuals and compared with the reported data of other populations.</p><p><b>RESULTS</b>The PowerPlex21 System had a power of discrimination (PD) ranging from 0.7839 to 0.9852 and a power of exclusion (PE) ranging from 0.2974 to 0.8099 for the 20 loci. No significant deviation from Hardy-Weinberg expectations was found for all the loci except for D5S818. This southern Chinese Han population had significant differences in the allele frequencies from 8 ethnic groups reported in China, and showed significant differences at 8 to 20 STR foci from 5 foreign populations. The allele frequency at the locus D1S1656 in this southern Chinese Han population differed significantly from those in the 5 foreign populations and from 3 reported Han populations in Beijing, Zhejiang Province and Fujian Province of China. The neighbor-joining phylogenetictree showed clustering of all the Asian populations in one branch, while the northern Italian and Argentina populations clustered in a separate branch. This southern Chinese Han population had the nearest affinity with the Yi ethnic population in Yunnan Province of China.</p><p><b>CONCLUSION</b>The 20 STR loci are highly polymorphic in this southern Chinese Han population, suggesting the value of this set of STR loci in forensic personal identification, paternity testing and anthropological study.</p>
Subject(s)
Humans , Asian People , Genetics , China , Cluster Analysis , Ethnicity , Forensic Sciences , Gene Frequency , Genetics, Population , Genotype , Microsatellite Repeats , Polymorphism, GeneticABSTRACT
<p><b>OBJECTIVE</b>To obtain two-dimensional gel electrophoresis maps of the prefrontal cortex (PFC) proteins of normal and heroin-addicted rats for identifying the differentially expressed proteins in the addicted rats.</p><p><b>METHODS</b>Rat models of heroin addiction were established, and the proteins in the PFC underwent two-dimensional gel electrophoresis with immobiline pH gradient isoelectric focusing as the first and vertical SDS-PAGE as the second dimension. ImageMaster 2D 5.0 analysis software, matrix assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) or MALDI-TOF-TOF-MS as well as NCBInr database searching were performed to separate and identify the proteome in the rat PFC.</p><p><b>RESULTS</b>Seven protein spots to represent 5 proteins were identified by PMF and MALDI-TOF-TOF-MS. Among those proteins, glucose-regulated protein (58 kD) and 26 S proteasome subunit p40.5 existed only in heroin-addicted rats, in which ATP synthase D chain was down-regulated. Ndufa10 and Eno1 were present in both groups, but their molecular mass and pI were different.</p><p><b>CONCLUSION</b>Immobilized pH gradient two-dimensional gel electrophoresis allows good reproducibility in separation and identification of the proteome in the PDF of heroin-addicted rats, which facilitates further investigation of pathogenic mechanisms of heroin addiction and central nerve injury.</p>