Objective To design the small interference RNA (siRNA) specific to human CCL20 gene by RNA interfering technique, construct its recombinant lentiviral expression vectors, and identify these vectors by DNA sequencing. Methods According to Tuschl’s principle, the siRNA was designed and converted into cDNA of shRNA (small hairpin RNA) of siRNA for CCL20 gene. The cDNA was synthesized and inserted into plasmid pHSER-dsRNA-GFP-SIN which was linearized by restriction endonucleases SpeⅠ and SalⅠ. The recombinant plasmid was transformed into competent E. coli. DH5? cells. The positive recombinant colony was selected by ampicillin medium agar and identified by DNA sequencing. Results Two recombinant lentiviral plasmids of siRNA specific to CCL20 gene were constructed successfully. Their DNA sequence analysis completely coincided with their designed sequences. Conclusion Lentiviral vector-based siRNA expression plasmids against CCL20 gene have been successfully constructed and identified. They will be further used for interfering CCL20 mRNA transcription and lay the foundation for CCL20 gene modified human keratinocyte stem cells.

Objective To explore the association between GNβ3 gene and depression,and to investigate the interaction of gene-environment(negative life events and childhood trauma) and potential possible pathogenesis of depression.Methods The sample of peripheral blood was collected from Chinese Northern patients(n=500) and controls(n=500).Snapshot technique was used to detect the genotype frequency and allele frequency of GNβ3 rs5443 polymorphism in cases and controls.The genotype and allele frequencies were analyzed by Chi-square test,the interactions of gene-environment were analyzed by Logistic regression.Results GNβ3 rs5443 genotype and allele frequencies were observed between patients and controls (x2 =20.249,P＜0.01;x2 =4.803,P＜0.05).There were genotype CC 102 and 158,genotype CT 280 and 217,genotype TT 118 and 125;allele C 484 and 533,allele T 516 and 467 between patients and controls,respectively.Logistic regression analysis showed that the interaction between rs5443 T+ and negative life events was associated with depression (P＜0.05,OR=1.957).In addition,individual carrying rs5443T+ genotypes and negative life events could increase risk of depression.Conclusion GNβ3 rs5443 is a possible susceptibility gene of depression.The interaction between rs5443 and negative life events is associated with depression.

Objective To investigate the single nucleotide polymorphisms(SNPs) of 5-HT1BR(rs6298),5-HT2AR(rs6311,rs6313) and 5-HT2BR(rs765458) gene,and their gene-gene interactions on suicidal behavior in major depressive disorder.Methods The blood samples were taken from 281 depression patients with impulsive suicide attempt and 281 age-matched healthy controls from a hospital in Harbin city,Heilongjiang province.The DNA isolated from blood samples and was genotyped using TapMan SNP genotyping probe.χ2 test was used to compare differences in the distribution of gene alleles between cases and controls.Haplotype and linkage disequilibrium(LD) analysis was performed using Haploview 4.0 software.GMDR was used to analyze the gene-gene interaction.Results The rs6313 and rs6311 of the 5-HT2AR gene were in strong linkage disequilibrium (D=0.756,r2=0.375).There was a significant gene-gene interaction of 5-HT1BR (rs6298),5-HT2AR (rs6311,rs6313) and 5-HT2BR(rs765458) on suicidal behavior(P<0.05).In this model,the test accuracy was 0.6182 and CV value was 10/10.Conclusion A haplotype containing rs6311 and rs6313 of 5-HT2AR gene is associated with suicidal behavior.The interaction of 5-HT1BR gene (rs6298),5-HT2AR gene (rs6311,rs6313) and 5-HT2BR gene (rs765458) are associated with depression suicidal behavior.