OBJECTIVETo study the suppressive effect of LRRC4 gene on human glioma U251 cells and further investigate its biological functions.

METHODSH&E, DNA and AgNORs stainings were performed on LRRC4-transfected U251 cells, mock-transfected U251 cells and non-transfected U251 cells, respectively. Quantitative analysis including cell morphometry, DNA content, DNA ploidy, silver stained argyrophilic nucleolar organizer regions (AgNORs) were investigated by image analysis. Flow cytometry was employed to determine the difference of cell cycle distribution and MTT staining was used to elucidate the activity of the LRRC4-transfected U251 cells.

RESULTSThe morphological cell parameters such as area, perimeter and diameter, DNA content, chromosomal aneupoloidy, mean area of AgNORs particles and mean nucleus area of the LRRC4-transfected U251 cells were remarkably decreased compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). Meanwhile, significant accumulation of cells in G(0)/G(1) phase but decrease of cells in S and G(2)/M phase, was observed in transfected U251 cells compared to those of the mock-transfected and non-transfected U251 cells (P < 0.05, P < 0.01). MTT staining showed that proliferation activity of both the mock- and non-trasfected U251 cells was significantly higher than that of the U251 cells transfected with LRRC4 gene (P < 0.01).

CONCLUSIONLRRC4 gene might be involved in tumor suppression by restraining DNA synthesis and the nucleoli organizer regions-associated proteins, keeping the cell cycles in phase G(0)/G(1) and reducing proliferation activity of the glioma cells. Morphometry combined with other techniques such as flow cytometry and MTT staining can well elucidate the biological function of novel genes.

Brain Neoplasms ; genetics ; pathology ; Chromosomes, Human, Pair 7 ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor ; physiology ; Glioblastoma ; genetics ; pathology ; Humans ; Nerve Tissue Proteins ; genetics ; physiology ; Transfection ; Tumor Cells, Cultured

Objective To investigate the psychological status of patients with brain concussion. Methods A total of 186 patients with brain concussion were evaluated using Symptom Check-List 90 (SCL-90) and the scores were compared with the norms. Results The general scores (137.71±39.48), total mean scores (1.53±0.44) and positive object number (32.90±19.41) of SCL-90, as well as the factor scores ofsomatization(1.57±0.52), compulsion(1.79±0.50), anxiety disorder(1.50±0.49), hostility(1.63± 0.57), phobic anxiety(1.57±0.51), paranoid ideation(1.62±0.51) and psychoticism ( 1.49±0.43) in the patients with brain concussion were all higher than the normal ones (129.96±38.76, 1.44±0.43, 24.92± 18.41, 1.37±0.48, 1.62±0.58, 1.39±0.43, 1.48±0.56, 1.23±0.41, 1.43±0.57, 1.29±0.42, respectively)(P< 0.05). The scores of interpersonal sensitivity (1.67±0.54) were also significantly higher than that of normal (P>0.05). Conclusion Brain concussion may lead to psychological disturbance, for which early interventions should be administered.

Objective To study the correlation between the changes of serum neuron specific enolase (NSE) level and psychological statuses in patients with cerebral concussion.Methods Forty patients with cerebral concussion,admitted to our hospital from January 2012 to December 2012,were chosen in our study as experimental group,and other 40 healthy controls performed physical examination in our Physical Examination Center at the same period were chosen.The serum NSE level in the two groups was determined by enzyme-linked immunosorbent assay (ELISA); Symptom Checklist 90 (SCL-90) scale was performed in 40 patients at different times after injury,and these data were compared with those from the national norm; the correlation between the changes of serum NSE level and SCL-90 scores was analyzed.Results The serum NSE level in the experimental group 1 d after the injury was obviously higher than that in the control group (P＜0.05); 3 and 7 d after the injury,the serum NSE level in the experimental group was close to normal level,which showed no significant differece as compared with that in the control group (P＞0.05).The SCL-90 scores in the experiemtal group 1,3 and 7 d after injury was signficantly higher than that of the national norm (P＜0.05).Positive corelation was noted between serum NSE level and SCL-90 scores 1 and 3 d after injury (r=0.498,P=0.001; r=0.418,P=0.007);however,negative corelation was noted btween the two 7 d after injury (r=0.213,P=0.186).Conclusion Different corelation can be noted between serum NSE level and SCL-90 scores at different time points;combined application of serum NSE level and SCL-90 scores can promote the diagnosis and treatment of patients with brain concussion.

Aim To study the effect of three kinds of active ingredients of traditional Chinese medicine (sinomenine, rhynchophylline and isorhynchophylline) on dre-miR-723-5p expression in morphine-induced zebrafish brain. Methods Morphine was injected intraperitoneally to zebrafish, conditional position preference (CPP) was trained and then the behavioral of animals were observed; the miRNA expression profiles of morphine-additive zebrafish were determined by small RNA sequencing; qRT-PCR was used to verify the expression of dre-miR-723-5p, three target gene databases (miRanda, miRDB, andRNAhybrid) were used to predict the target genes of dre-miR-723-5p; Kobas 3.0 was used to perform Gene Ontology (GO) and KEGG pathway analysis of these target genes. Results Morphine-induced CPP model was established successfully. Compared with control group, the resident time and movement map in drug-pair box of zebrafish in model group significantly increased. After drug administration, the resident time and movement map in drug-pair box of zebrafish decreased. The verification results of qRT-PCR were consistent with the results of small RNA sequencing. Ninety-nine putative target genes of dremiR-723-5p that were common to all three target gene databases, which were mainly enriched in biological process, cell composition and molecular function, involved in the positive regulation of MAPK signaling pathway, lysosome, cytokine-cytokine receptor interaction, and apoptosis. Conclusion Morphine can increase the expression of dre-miR-723-5p in the zebrafish brain, which can be reversed by sinomenine, isorhynchophylline, and rhynchophylline treatment, and dre-miR-723-5p may participate in the mechanism underlying morphine-induced damage of brain.