Seeking for potential pathogenic genes of major depressive disorder in the Gene Expression Omnibus database.

INTRODUCTION: Major depressive disorder (MDD) is one of the most common mental disorders worldwide. The aim of this study was to identify potential pathological genes in MDD. METHODS: We searched and downloaded gene expression data from the Gene Expression Omnibus database to identify differentially expressed genes (DEGs) in MDD. Then, Kyoto Encyclopedia of Genes and Genomes pathway, Gene Ontology analysis, and protein-protein interaction (PPI) network were applied to investigate the biological function of identified DEGs. The quantitative real-time polymerase chain reaction and a published dataset were used to validate the result of bioinformatics analysis. RESULTS: A total of 514 DEGs were identified in MDD. In the PPI network, some hub genes with high degrees were identified, such as EEF2, RPL26L1, RPLP0, PRPF8, LSM3, DHX9, RSRC1, and AP2B1. The result of in vitro validation of RPL26L1, RSRC1, TOMM20L, RPLPO, PRPF8, AP2B1, STIP1, and C5orf45 was consistent with the bioinformatics analysis. Electronic validation of C5orf45, STIP1, PRPF8, AP2B1, and SLC35E1 was consistent with the bioinformatics analysis. DISCUSSION: The deregulated genes could be used as potential pathological factors of MDD. In addition, EEF2, RPL26L1, RPLP0, PRPF8, LSM3, DHX9, RSRC1, and AP2B1 might be therapeutic targets for MDD.

Transcatheter closure of perimembranous ventricular septal defect using a modified double-disk occluder.

This multicenter study assessed the efficacy and safety of transcatheter closure of perimembranous ventricular septal defect (VSD) using a modified double-disk occluder. In 5 different centers in China, 412 patients with VSD, including 202 men and 210 women, underwent attempted transcatheter closure. The age range was 3 to 65 years (mean 16.4+/-9.1). The diameter of defect was 3 to 15 mm by transthoracic echocardiography and 3 to 18 mm by left ventriculography. The ratio of pulmonary to systemic flow varied from 1.6 to 2.3 (1.9+/-0.4). The device diameter was 4 to 20 mm (7.09+/-3.60). The ventricular septal rim below the aortic valve was 0 to 5 mm. The immediate success rate was 96.6%; 6 cases had third degree atrioventricular block and recovered within 3 weeks. None needed a permanent pacemaker. Dislodgement of the device occurred in 3 patients but the device was recaptured and redeployed in 2 cases. During the follow-up period of 2 years, there was no evidence of residual shunt and device-related complications. In conclusion, transcatheter closure of VSD is safe and effective in most selected patients; the mid-term prognosis of patients with transcatheter closure is good.

[Cloning expression and characterization of a multifunctional anticoagulated peptide gene in E. Coli].

OBJECTIVE: To investigate the feasibility about the cloning, expression and characterization of multifunctional anticoagulated peptide. METHODS: We designed a construct using glutathione S-transferase (GST) as a protein vector, fused via a cleavable linker to an antithrombotic peptide of 31 amino acids. The peptide was designed to include three inhibitory regions: (1) the Lys-Gly-Asp (KGD) amino acid sequence to prevent fibrinogen binding to platelets; (2) a part of fibrinopeptide A, an inhibitor of thrombin; and (3) the tail of hirudin, a potent direct antithrombin. The amino acid sequence of the 31 amino acid peptide was reverse translated, and the gene was chemically synthesized and cloned into an expression vector pGEX-5X-3 as a 3'fusion to the GST gene. Gene expression was induced in E. coli DH5 alpha cells and the fusion protein was purified using affinity chromatography. RESULTS: The purified fusion protein significantly lengthened the activated partial thromboplastin time (74.7 s tested in 80 micromol/L) and thrombin time (102.3 s tested in 80 micromol/L) and inhibited the amidolytic activity of thrombin 11% activity compared with the control tested in 100 micromol/L. The ADP-induced platelet aggregation was markedly inhibited by the purified fusion protein. The study has also shown that GST exhibits relevant activity of antiplatelet weaker than the purified fusion protein. 20.7% activity compared with the control tested in 100 micromol/L. CONCLUSION: Our results confirm that it is feasible to design a hybrid multifunctional protein that targets various components of the haemostatic process.

[Expression and purification of recombinant glycoprotein (GP) IIb/IIIa receptor antagonists].

To investigate the effect of GST-KGDX (glutathione S-transferase-Lys-Gly-Asp-X) fusion protein, GP IIb/IIIa receptor antagonist, on platelet function in vitro. The KGDX (Lys-Gly-Asp-X) gene was assembled from 2 synthetic oligonucleotides, 36 bp in length, using BamH I and Xho I restriction enzyme sites at the end of the gene for cloning into the expression vector pGEX4T-1. Expression of fusion protein was directed by the tac promoter. The Escherichia coli DH5a contained the plasmid pGEX-4T-1-KGDX was expressed by 37 degrees C heat induction. The fusion protein of KGDX with glutathione S-transferase (GST-KGDX) was purified in one step from the bacterial lysate by glutathione-agarose beads for affinity chromatography. GST-KGDX was found to be soluble and abundant, the yield of 35 mg/L of cultures was obtained. The GST-KGDX was expressed in E. coli to a level of 48.02% of total cellular protein. GST-KGDX inhibited ADP-induced human platelet aggregation stronger than GST (P < 0.05 or < 0.01). In flow cytometry assay for fibrinogen binding, both GST and GST-KGDX inhibited platelet aggregation by binding with high affinity to GPIIb/IIIa. Mean fluorescence intensity of GST-KGDX fusion protein was significantly higher than that of GST. It is concluded that the GST-KGDX fusion protein can be produced by E. coli and used as an antiplatelet agent.