CCR5 serves as an essential coreceptor for human immunodeficiency virus type 1 (HIV-1) entry, and individuals with a CCR5 D32 variant appear to be healthy, making CCR5 an attractive target for control of HIV-1 infection. The CRISPR/Cas9, which functions as a naturally existing adaptive immune system in prokaryotes, has been recently harnessed as a novel nuclease system for genome editing in mammalian cells. Although CRISPR/Cas9 can be readily delivered into cell lines, due to the large size of the Cas9 protein, efficient delivery of CCR5-targeting CRISPR/Cas9 components into primary cells, including CD4 + T-cells, the primary target for HIV-1 infection in vivo, remains a challenge. In the current study, following design of a panel of top-ranked single-guided RNAs (sgRNAs) targeting the ORF of CCR5, we demonstrate that CRISPR/Cas9 can efficiently mediate the editing of the CCR5 locus in cell lines, resulting in the knockout of CCR5 expression on the cell surface. Next-generation sequencing revealed that various mutations were introduced around the predicted cleavage site of CCR5. For each of the three most effective sgRNAs that we analysed, no significant off-target effects were detected at the 15 top-scoring potential sites. More importantly, by constructing chimeric Ad5F35 adenoviruses carrying CRISPR/Cas9 components, we efficiently transduced primary CD4 + Tlymphocytes and disrupted CCR5 expression, and the positively transduced cells were conferred with HIV-1 resistance. To our knowledge, this is the first study establishing HIV-1 resistance in primary CD4 + T-cells utilizing adenovirus-delivered CRISPR/Cas9.

The "Learn to Think" (LTT) Intervention Program was developed for raising thinking abilities of primary and secondary school students. It has been implemented in more than 300 schools, and more than 200,000 students took part in the experiment over a 10-year span. Several longitudinal intervention studies showed that LTT could promote the development of students' thinking ability, learning motivation, and learning strategy as well as raise academic performance in primary schools. This article describes a study of the influence and the delayed effects of LTT on the scientific creativity of secondary school students. One hundred and seven students were selected from a secondary school, 54 of them participated in the LTT every 2 weeks and the rest had not. The intervention lasted 2 years, and the delayed effect was explored half a year after terminating the intervention. The Scientific Creativity Test for Secondary School Students was used four times from pre-test to delayed post-test. The results indicated that the LTT did promote the development of scientific creativity of secondary school students, and the effects on the scientific creativity were not necessarily immediate, but tended to be long-lasting.

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