Rapid construction of phosphatase and tensin homolog-deleted on chromosome ten gene recombinant adenovirus using the AdEasy system

Recent studies have shown that phosphatase and tensin homolog-deleted on chromosome ten （PTEN） gene plays an important role in ischemic brain damage and synaptic plasticity. The AdEasy system, which has been widely used, greatly simplifies preparation of recombinant adenovirus. Therefore, recombinant defective adenovirus vector carrying human PTEN tumor suppressor gene （Ad-PTEN） was constructed using the AdEasy-1 system and was transfected into HEK293 cells for packaging and amplification. Infection efficiency and expression intensity were observed in primary cultured rat hippocampal neurons infected with Ad-PTEN in vitro. Results revealed a cytopathic effect in green fluorescent protein expression, which increased with prolonged time. After three cycles of amplification, the adenovirus titer was increased to an adequate titer for infecting hippocampal neurons. The entire process typically requires 4-5 weeks for completion. Results suggested that recombinant defective adenovirus vector carrying the PTEN gene was successfully and rapidly constructed using the AdEasy system.

A Probe into the Implication of Education with Zhang Junmai’s Philosophy of Mind

In modern times,under the dual crisis of nationality and culture,Zhang Junmai returned to the internal philosophy of“Inner Sage,”whereby he started the global exploration of“Outer King”and constructed his own categorized Neo-Confucianism thoughts based on the contents of Yangming’s philosophy of mind as well as western philosophies.Therefore,by probing into the foundation of Zhang Junmai’s philosophy of mind,this paper explores the educational significance of the view of life based on honesty,the methodology of giving equal importance to learning and doing,and the world’s point of view contained in it,which may serve as a certain reference for the current education development.

Genetic investigation of haemoglobinopathies in a large cohort of asymptomatic individuals reveals a higher carrier rate for b-thalassaemia in Sichuan Province (Southwestern China)

The incidence of haemoglobinopathy is high in China,especially south of the Yangtze River.However,the exact status of haemoglobinopathy in Sichuan is unknown.To carry out a detailed research of haemoglobinopathy in individuals living in Sichuan,13,298 subjects without clinical symptoms who were living in Sichuan Province,with an age distribution of 5e73 years,were included in this study.Between March 2014 and July 2017,these subjects received examinations at the Medical Lab of Chengdu Women’s&Children’s Central Hospital.Mean corpuscular volume(MCV)<82 fL or mean corpuscular haemoglobin(MCH)<27 pg was used to indicate haemoglobinopathy carriers.Abnormal haemoglobin was screened by electrophoresis,and genes were sequenced to identify genotypes.Genotype diagnosis of alpha-and beta-thalassaemia was carried out by using PCR and shunt hybridization.There were 638 suspected haemoglobinopathy carriers(4.80%,638/13,298).DNA sequencing identified 6 subjects with abnormal haemoglobin genotypes and 15 subjects with Hb E.The frequency of heterozygosity for thalassaemia was 4.12%(1.48%for α-thalassaemia and 2.61%for β-thalassaemia)in Sichuan Province.The mutation spectrum of α-thalassaemia consisted of the five most common mutations:–^(SEA),-α^(3.7),-α^(4.2),α^(CS),and α^(QS).Seven types of β-thalassaemia mutation were found in this study:CD41-42(-TTCT)was the most frequent(28.47%),followed by 17(A>T),28(A>G),and IVS-Ⅱ-654(C>T).The main abnormal haemoglobin genotype(HbE)and thalassaemia genotype(–^(SEA),CD41-42(-TTCT))were consistent with those in other regions of China,but the carrier rate of β-thalassaemia in Sichuan was higher than that of α-thalassaemia.