ObjectiveTo construct expressing vector of microRNA with molecular cloning methods which target CD4 and electroporating the vector to the MT4 cell to determine its effect to CD4 expression.MethodsPredict a microRNA can target CD4.Linking the pre-mir-181a PCR products to T vector,then cloning into the pEGFP-N1 vector after enzyme digestion.To test the integrity through the colony PCR and sequencing analysis.Electroporating the vector to MT4 cell,using FACS to test the CD4 expression.ResultsHsa-mir-181a is able to target CD4.The sequence of the construct was correct.The hsa-mir-181a is stable expressing in MT4 after electroporating with the vector and MT4 cell CD4 was down-regulated.ConclusionThe construct can be stable expressing hsa-mir-181a in MT4 cell and down-regulating the CD4 expressing.This method can be utilized as a novel intervention to the HIV fusion,it shows potential as a gene therapy tool in vitro.

BMP6 is a potent protein for future treatment strategies of bone regeneration as it is a very important regulator of bone homeostasis. Active BMP6 is a dimer containing multidisulfide bonds and is a highly hydrophobic protein prone to aggregation. To obtain soluble and active BMP6 in Escherichia coli, we compared the effects of four N-terminal fusion tags (TRX, GST, MBP and CBD) and N-terminal His6-tag. The expression and solubility were tested under the different conditions (expression hosts, temperatures and inductor concentrations). A series of experiments leads to the finding that the placement of MBP before the BMP6 is best in availing the soluble expression of the protein. Our study alsodemonstrates that in E. coli BL21trxB(DE3) cytoplasm, which is a thioredoxin reductase mutant strain, soluble homodimeric BMP6 can be formed. The overexpressed MBP-BMP6 fusion protein is purified by chromatography, and shown to be functionally active.
Bone Morphogenetic Protein 6 ; biosynthesis ; genetics ; Carrier Proteins ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Maltose-Binding Proteins ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; metabolism ; Solubility ; Transformation, Bacterial

OBJECTIVETo assess the value of chromosome microarray analysis (CMA) for identifying the etiology of developmental delay/intellectual disability (DD/ID).

METHODSA total of 489 children referred for DD/ID with or without other abnormalities were recruited. All patients showed a normal karyotype. DNA was extracted and hybridized with Affymetrix CytoScan 750K array by following the manufacturer's protocol. The data was analyzed with CHAS v2.0 software.

RESULTSThe children were classified as with isolated DD/ID (n=358), DD/ID with epilepsy (n=49), and DD/ID with other structural anomalies (n=82). Pathogenic copy number variants (CNVs) were identified in 126 cases (25.8%), which included 89 (24.9%, 89/358) of whose with isolated DD/ID, 13 (26.5%, 13/49) of those with DD/ID and epilepsy, and 24 (29.3%, 24/82) of whose with DD/ID and other structural anomalies [P=0.064 (24.9% vs. 26.5%), P=0.679 (24.9% vs. 29.3%), and P=0.113 (26.5% vs. 29.3%), respectively]. Among the 126 cases, 79 were identified as microdeletion/microduplication syndromes, which included 15q24 microdeletion syndrome, Xq28 microduplication syndrome, and Lowe syndrome. Forty-seven cases had de novo pathogenic CNVs. ABAT, PMM2, FTSJ1, DYNC1H1 and SETBP1 were considered as candidate genes for DD/ID.

CONCLUSIONCMA is an effective method for identifying the etiology of DD/ID and is capable of identifying microdeletion/microduplication syndromes as well as de novo pathogenic CNVs which may be missed by conventional karyotyping. Based on the results, candidate genes for DD/ID may be identified.

Child ; Child, Preschool ; Chromosomes ; genetics ; Developmental Disabilities ; genetics ; Female ; Humans ; Infant ; Intellectual Disability ; genetics ; Karyotyping ; methods ; Male

Objective:To investigate the effectiveness of case-based learning (CBL) combined with online teaching in standardized residency training of rheumatology and immunology.Methods:A total of 78 individuals who participated in standardized residency training in Department of Rheumatology and Immunology in our hospital from June 2019 to August 2020 were included and divided into observation group and control group. The individuals in the control group received traditional teaching, and those in the observation group received CBL combined with online teaching. The physicians receiving standardized residency training were evaluated by theoretical examination, clinical operation skill assessment, and instructor rating, and the degree of satisfaction with teaching, degree of satisfaction with teaching methods, and classroom learning atmosphere were also evaluated.Results:The observation group had a theoretical examination score of (94.10±2.01) and a clinical operation skill assessment score of (90.44±1.57), which were significantly higher than those of the control group ( P<0.05), and the observation group had a significantly better instructor rating (89.36±1.33) than the control group ( P<0.05). Compared with the control group, the observation group had significantly higher degree of satisfaction with teaching (3.79±0.41), degree of satisfaction with teaching methods (3.92±0.27), and evaluation of classroom learning atmosphere (3.90±0.31) ( P<0.05). Conclusion:CBL combined with online teaching can help to improve learning efficiency, stimulate the enthusiasm for learning, expand clinical thinking, promote the growth of teaching and learning, and form a virtuous cycle among trainees receiving standardized residency training, which holds promise for further exploration.