Application of Metabonomics in Substance Abuse Toxicology Research.

Abstrct: Metabonomics is a relative discipline that develops after genomics and proteomics, and it is an important component of systems biology. It uses high-throughput and high-sensitivity instruments to perform qualitative and quantitative analysis of all metabolic components in specific biological samples under limited conditions and combines with multivariate statistics to analyze and process the data to obtain information about physiological, pathological or toxicological changes in organisms. In recent years, because of the complicated mechanism of substance abuse and the continuous emergence of new psychoactive substances, metabonomics is increasingly used in substance abuse research. Therefore, this article reviews the application of metabonomics of substance abuse in the toxic mechanism, the mechanism of addiction and the discovery of biomarkers.

Analysis of silent information regulator 1 (SIRT1) gene polymorphisms in antituberculosis- drug-induced hepatotoxicity in a prospective cohort study.

OBJECTIVE: Antituberculosisdrug-induced hepatotoxicity (ATDH) is a common and sometimes serious side effect related to tuberculosis (TB) treatment. A number of risk factors and host genetics contribute to the development of ATDH. However, genetic factors of ATDH remain to be identified. Silent Information Regulator 1 (SIRT1), an essential metabolism gene, was proved to be involved in ATDH in mice. The aim of this investigation was to study the association between ATDH and tag-single nucleotide polymorphisms (tag-SNPs) of the SIRT1 gene in a prospective cohort study in patients with TB. METHODS: 280 newly diagnosed TB patients were recruited in this study before starting first line anti-TB treatment and were followed up for 3 months after initiating anti-TB therapy. The tag-SNPs were selected by using Haploview 4.2 based on the HapMap database of Han Chinese Beijing. Genotyping was performed by polymerase chain reaction (PCR) and the Sequenom MassARRAY iPLEX platform. RESULTS: 24 (9.8%) of the 245 patients included in the final analysis developed hepatotoxicity during the following up period. No significant differences in the allele, genotype, or haplotype frequency distributions of the tag- SNPs (rs7069102, rs2273773, rs4746720) of the SIRT1 gene were identified between the ATDH and non-ATDH groups (all p > 0.05). CONCLUSIONS: The SIRT1 gene may not contribute to the risk for developing hepatotoxicity during anti-TB treatment in the Han Chinese population.

Molecular cloning and prokaryotic expression of vp5 gene of grass carp reovirus strain GCRV096.

VP5 is an outer capsid protein of grass carp reovirus (GCRV). It is predicted to involve in helping GCRV enter the host cells. In this study, the full-length vp5 gene (accession number in GenBank: JN206664.1) was cloned from GCRV strain GCRV096, which was isolated from diseased grass carp (Ctenopharyngodon idella) in southern China by RT-PCR technique using the primers designed from the known vp5 gene sequences of other strains of GCRV published in GenBank. The ORF sequence of vp5 is composed of 1,947 nucleotides encoding a 648-residues protein with a calculated molecular mass of 68.6 kDa and an estimated isoelectric point of 6.1. Sequence analysis results showed that VP5 might serve as a penetration protein and play an important role in GCRV penetration into the host cells. A full length of vp5 gene was subcloned into the prokaryotic expression vector pET-28a (+) and the recombinant plasmid (pET/GCRV-VP5) was then transduced into Escherichia coli BL21 (DE3) cells to express VP5 in vitro. SDS-PAGE and western blotting analysis indicated that the protein expressed successfully. Results also showed that the fusion protein expressed in the form of inclusion body, and it expressed in the highest level when induced with 0.2-mM IPTG at 28 °C for 4 h. These results are important for the future study on the molecular structure, function, and immunogenicity of GCRV capsid protein.

Analysis of the genetic diversity of the lymphocystis virus and its evolutionary relationship with its hosts.

Lymphocystis disease virus (LCDV) is the causative agent of lymphocystis disease. In this study, the mcp gene of LCDV and the cyt b gene of the host fish were selected as molecular markers, and the phylogenetic relationships between LCDV and its host were analyzed. The 25 LCDV isolates examined in this study were attributed to seven LCDV genotypes: genotype I (LCDV-1), genotype II (LCDV-cn, etc.), genotype III (LCDV-rf), genotype IV (LCDV-rc and LCDV-sb), genotype V (LCDV-cb), genotype VI (LCDV-tl), and genotype VII (LCDV-sa). Genotype VII is a new genotype. LCDV1 was found to have differentiated first, followed by LCDV-rf; then LCDV-tl; LCDV-cb; and then LCDV-sa; and by LCDV-rc and LCDV-sb; and finally by LCDV-cn, LCDV-C, and LCDV-jf. From the host evolutionary perspective, Rachycentron canadum was found to have differentiated first, followed by Trichogaster leeri, Chanda baculis, and Sebastes schlegeli, Lateolabrax sp., Sparus aurata, Platichthys flesus, and Paralichthys olivaceus. Comparison of the phylogenies of the host fish species and LCDVs revealed no significant evidence of cospeciation between LCDVs and their host fish. In-depth studies of the genetic variation in LCDVs can enhance our understanding of the mechanism of LCDV infection, which may provide important insights into the prevention and treatment of lymphocystis disease.

Phylogenetic analysis of newly isolated grass carp reovirus.

Grass carp reovirus (GCRV) is a causative agent of haemorrhagic disease in grass carp that drastically affects grass carp aquaculture. Here we report a novel GCRV isolate isolated from sick grass carp that induces obvious cytopathic effect in CIK cells and name it as GCRV096. A large number of GCRV 096 viral particles were found in the infected CIK cells by electron microscope. The shape, size and the arrangement of this virus were similar to those of grass carp reovirus. With the primers designed according to GCRV 873 genome sequences, specific bands were amplified from sick grass carp and the infected CIK cells. The homology rates among vp4, vp6 and vp7 gene in GCRV 096 and those of some GCRV isolates were over 89%. In this study, the sequences of vp4, vp6 and vp7 were used to analyse sequence variation, phylogenetic relationships and genotypes in twenty five GCRV isolates. The results indicated these twenty five GCRV isolates should be attributed to four genotypes. And there were no obvious characteristics in the geographical distribution of GCRV genotype. The study should provide the exact foundation for developing more effective prevention strategies of grass carp haemorrhagic disease.

[Molecular phylogenetic relationships of Drosophila immigrans species group based on H2a-H2b spacer and Cyt b sequences].

In order to investigate molecular phylogenetic relationships of Drosophila immigrans species group, nucleotide sequences of H2a-H2b spacer regions and partial nucleotide sequences of Cytochrome b gene for five specieses subgroup in Drosophila immigrans species group and D. hydei which was selected as outgroup were acquired. Phylogenetic trees were reconstructed using maximum parsimony, maximum likelihood, neighbor joining and Bayesian methods based on H2a-H2b and Cyt b DNA sequences. The results indicated that Drosophila curviceps species subgroup is located at the base of molecular trees, diverged first in this group; the second clade is Drosophila quadrilineata species subgroup; the third is Drosophila hypocausta species subgroup; the fourth is Drosophila immigrans species subgroup; Drosophila nasuta species subgroup is the last diverged clade. Moreover, the phylogentic relationships of seven specieses in Drosophila nasuta subgroup was investigated.