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1.
Chinese Journal of School Health ; (12): 1353-1355, 2019.
Article in Chinese | WPRIM | ID: wpr-816794

ABSTRACT

Objective@#To re-evaluate the reliability and validity of the Adolescent Self-rating Life Events Checklist (ASLEC), and to adapt to the further application to middle school students in rural China.@*Methods@#A stratified random cluster sampling method was applied to select 15 607 adolescents from grade 7th to grade 12th in 15 rural areas of 5 provinces(Anhui Province, Yunan Province, Guangdong Province, Hei Longjiang Province, Hubei Province), and they were recruited to complete our Questionnaires.@*Results@#The revised version (ASLEC-R) consisted of 5 dimensions (punishment, interpersonal relationship, academic pressure, loss and adaptation problem), 25 items after deleting items 5 and 17 through exploratory factor analysis and confirmatory factor analysis, which could accounted for 55.22% of the total variance. The fit indices were RMSEA=0.06,GFI=0.91,CFI=0.88,TLI=0.86,NFI=0.88,AGFI=0.88,HOELTER 0.05=261. The Cronbach’s α and Spearman-Brown splithalf reliability coefficient of the whole scale were 0.92 and 0.87, respectively,and the test-retest reliability was 0.84. ASLEC-R had better reliability than the unrevised version. The results of five-joint item analyses showed that each item improved in terms of indiscrimination, relevance, contribution, homogeneity and sensitivity. The correlation coefficients with BWAQ and EI subscale were 0.38 and -0.36 respectively.@*Conclusion@#ASLEC-R has good reliability and validity , and it is worth being applied to the Chinese rural areas.

2.
Electron. j. biotechnol ; 19(4): 75-80, July 2016. ilus
Article in English | LILACS | ID: lil-793956

ABSTRACT

Background: Using recombinant adeno-associated virus 2 (rAAV-2), we attempted to establish a HEK293T cell line that is able to site-specifically integrate and stably express glial cell line-derived neurotrophic factor (GDNF). Results:Recombinant vector with enhanced green fluorescent protein (EGFP) and GDNF (pTR-P5-EGFP-IRES-GDNF), as well as that carrying Rep genes and SV40 promoters (pSVAV2) were constructed and packed. HEK293T cells were co-infected with rAAV-2/EGFP-GDNF and rAAV-2/SVAV2 virus separately at 1 x 10(4),1 x 10(5),and 1x10(6) of multiplicity of infection (MOI). The efficiency of transduction was detected using flow cytometry. Additionally, the infected HEK293T cells were separately validated by touchdown polymerase chain reaction (PCR) and Western-blot. After 72 h of transduction, the rate of EGFP positive cell was 22%, 45% and 49% at the MOIs of 1 x 10(4),1 x 10(5) and 1 x 10(6), respectively. On the 3rd, 6th and 9th day of cell passage, there was no significant difference in the cell viability and proliferation rate between transduction and control groups. Importantly, touchdown PCR showed that there was a specific PCR amplified product band in the lane of infected cells. Furthermore, GDNF expression was detected in the infected cells after 15 and 180 d of cultivation. Conclusions: A HEK293T cell line able to site-specifically integrate and stably express GDNF was established.


Subject(s)
Dependovirus , Glial Cell Line-Derived Neurotrophic Factor , HEK293 Cells , Recombination, Genetic , Transduction, Genetic , Cell Line , Polymerase Chain Reaction , Green Fluorescent Proteins , Genetic Vectors , Microscopy, Fluorescence
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