Identification of Down syndrome fetal encephalopathy related genes and signaling pathways via bioinformatics analysis / 中华实用儿科临床杂志

Objective:To identify Down syndrome (DS) fetal encephalopathy related genes and signaling pathways via bioinformatics analysis, and to explore their potential mechanisms underlying the occurrence and development of DS neuropathology.Methods:Retrospective study.In December 2021, dataset GSE59630 was downloaded from the gene expression omnibus (GEO), and differentially expressed genes (DEGs) between DS and normal fetal brain tissue were identified by R software.Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and gene set enrichment analysis (GSEA) were performed on the genes identified.The protein-protein interaction (PPI) network was constructed based on search tool for the retrieval of interacting genes online database and Cytoscape software, and key modules and hub DEGs were identified.Real-time quantitative polymerase chain reaction technique was used to verify the expression of hub genes related to neurodegeneration in brain tissue of 3 pairs of DS and normal fetuses at the gestational age of 22-33 weeks.Results:A total of 225 DEGs were screened out from DS and normal fetal brain tissue, including 18 up-regulated genes and 207 down-regulated genes.GO functional enrichment analysis showed that DEGs were mainly enriched in neurogenesis, neuronal apoptosis, transcriptional regulation, mitochondrial energy metabolism, etc.KEGG pathway enrichment analysis revealed that DEGs were associated with a variety of neurodegenerative diseases.GSEA suggested that apoptosis and inflammatory responses play a vital part in the occurrence of DS neuropathology.Ten hub genes were identified by the PPI network established, and they were mainly related to histone acetylation and transcriptional regulation.According to the tissue verification result, the variations of RAB8A, TBP and TAF6 expression conformed to the microarray data. Conclusions:The key genes and signaling pathways identified by transcriptome analysis of fetal brain tissue facilitate the comprehensive understanding of the molecular mechanism of DS neuropathology.This study provides a novel insight into the clinical diagnosis and treatment of neurodevelopmental abnormalities and mental retardation in DS.

Receptor-interacting protein 140 overexpression promotes neuro-2a neuronal differentiation by ERK1/2 signaling / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Abnormal neuronal differentiation plays an important role in central nervous system (CNS) development abnormalities such as Down syndrome (DS), a disorder that results directly from overexpression of genes in trisomic cells. Receptor-interacting protein 140 (RIP140) is significantly upregulated in DS brains, suggesting its involvement in DS CNS development abnormalities. However, the role of RIP140 in neuronal differentiation is still not clear. The current study aimed to investigate the effect of RIP140 overexpression on the differentiation of neuro-2a (N2a) neuroblastoma cells, in vitro.</p><p><b>METHODS</b>Stably RIP140-overexpressing N2a (N2a-RIP140) cells were used as a neurodevelopmental model, and were constructed by lipofection and overexpression validated by real-time polymerase chain reaction and Western blot. Retinoic acid (RA) was used to stimulate N2a differentiation. Combining the expression of Tuj1 at the mRNA and protein levels, the percentage of cells baring neurites, and the number of neurites per cell body was semi-quantified to determine the effect of RIP140 on differentiation of N2a cells. Furthermore, western blot and the ERK1/2 inhibitor U0126 were used to identify the specific signaling pathway by which RIP140 induces differentiation of N2a cells. Statistical significance of the differences between groups was determined by one-way analysis of variance followed by the Dunnett test.</p><p><b>RESULTS</b>Compared to untransfected N2a cells RIPl40 expression in N2a-RIP140 cells was remarkably upregulated at both the mRNA and protein levels. N2a-RIP140 cells had a significantly increased percentage of cells baring neurites, and numbers of neurites per cell, as compared to N2a cells, in the absence and presence of RA (P < 0.05). In addition, Tuj1, a neuronal biomarker, was strongly upregulated in N2a-RIP140 cells (P < 0.05) and phosphorylated ERK1/2 (p-ERK1/2) levels in N2a-RIP140 cells were dramatically increased, while differentiation was inhibited by the ERK1/2-specific inhibitor U0126.</p><p><b>CONCLUSIONS</b>RIP140 overexpression promotes N2a cell neuronal differentiation by activating the ERK1/2 pathway.</p>

Exploration on a New Mode of training aid on healthcare talents of Western China by first class hospital / 中华医学科研管理杂志

Objective To promote professional training aid on healthcare talents in western China,this paper aims to explore methods and to evaluate the effectiveness of a new training system for underprivileged medical professionals from hospitals of western China.Methods Questionnaires and interviews were conducted to 115 trainees and their employers took part in the Counterpart Support Project.A retrospective analysis was done to evaluate key indicators of the trainees' professional competence.Results More than 90％ of the trainees have seen great improvement on basic healthcare knowledge,clinical skills,and diagnosis and treatment practice.About 80％ the participating hospitals have enjoyed varied degrees of growth in outpatient visits,surgical volume,number of inpatient and operational efficiency.(Increase level from 0％ to 50％) Conclusions Within the frist 3 years of implementation,this new model has been proven effective to enhance participants' overall professional competence,and by tapping into the significant resource of first class hospitals,this model will help greatly promoting the development of healthcare talents in Western China.

Reasons for encounter and diagnosis of pediatric patients at general practice clinic affiliated to a tertiary hospital / 中华全科医师杂志

This study reviewed the reasons for encounter (RFE),diagnoses and the diagnostic categories of patients aged 0-14 y in general practice clinic affiliated to a tertiary hospital from June 2011 to May 2012.The results showed that the most frequent RFE was cough,at a rate of 29.6 per 100 encounters;the second one was fever (21.4％) and the third was nasal problems (9.5％).Medical examinations,skin rashes and follow-up encounters occurred at equal rates (5.2％).The most frequent diagnosis was acute upper respiratory tract infection (41％).Medical examination (6.3％),other viral diseases (5.1％),pneumonia (3.6％) and no detected disease (3.6％) were also common diagnoses.The diagnostic categories included all systems,and the problems of the respiratory system (56.4％) were most common.The results suggest that pediatric care comprises a very important part of primary care.

Cloning,identification,and cellular localization of a down-regulated gene fragment related with Down Syndrome / 北京大学学报(医学版)

Objective: To clone a novel gene and explore its expression patterns in tissues and cells,so as to find its role in the process of encephalopathy in DS.Methods: On the base of our previous microarray's result together with the tissue type,we chose EST AI480014 to carry out RACE,then analyzed its expression profiles in liver,spleen,kidney,heart,brain by multi-tissues Northern blot,after that semi-quantitive RT-PCR was used to reexamine the expression profiles.Furthermore,we used ISH to find whether aim gene expressed in neuroglial cells cultured in vitro.Finally we performed semi-quantitive RT-PCR to explore whether it expressed differently between DS and normal.Results: We gained a 682 bp new cDNA fragment(DQ275636)which expressed in all the tissues examined and had no alternative splices in them.It expressed highly in brain especially in frontal lobe and hippocampus.According to the ISH result we convinced that it expressed in neuroglial cells.Using bioinformatics we mapped DQ275636 to chromosome 5q14.Conclusion: We have obtained a new gene fragment based on the(above) results.According to its expression character and tissue type,it can be suggested that this gene has a probable role in the process of encephalopathy in DS.

Cell-cycle negative regulatory gene ANA is over-expressed in the brain tissues of patients with Down syndrome / 北京大学学报(医学版)

Objective: To investigate the expression level of genes located in chromosome 21 in the brain tissues of Down syndrome(DS). Methods: An optimized semi-quantitative RT-PCR method was used to evaluate the expression levels of seven genes encoded in chromosome 21 in fetal cortex brain and cerebellum of DS and the control at the end of 20 weeks of gestation. B2M was used as internal reference to normalize cell loss. Results: The expression levels of 6 genes in cortex and cerebellum, including DYRK1A, SYNJ1, PCP4, C21orf5, C21orf2 and C21orf106, were comparable between DS and the control. ANA, a cell-cycle negative regulatory gene, was over-expressed dramatically in the cortex but not in the cerebellum of DS. Conclusion: Over-expression of ANA may contribute to the reduction of neuronal density in DS brain.

Preliminary exploration of the influence factors of degenerate oligonucleotide primered PCR of genome DNA / 北京大学学报(医学版)

Objective: To explore the influence factors of the degenerate oligonucleotide primered PCR(DOP-PCR). Methods: Genome DNA template from the mouse single oocyte or liver tissue were used to perform DOP-PCR. DOP-PCR was carried out with templates of different origin, different gradient dilution, with or without low melting point gel purified to wipe off the small fragment that might interfere with the following analysis, and then PCR of gene FTCD and CBS were carried out to evaluate the influence of these factors on the amplification efficiency and specificity. Results: Compared with genome DNA template from mouse liver, the template from single oocyte had the same efficiency and specificity but a minor yield and different gradient dilution of DNA template had no effect on the efficiency and specificity. Furthermore, there was a higher specificity in the low melting point gel-purified DOP-PCR product than in untreated ones. Conclusion: We have got a satisfactory result and increased specificity from DOP-PCR product purified with the low melting point gel. Single oocyte of mice could be used for further investigation of special genes detection by DOP-PCR and of an optimization in the yield of the products.