[Construction of recombinant adenovirus expressing shRNA targeting human leukocyte-derived arginine aminopeptidase gene].

Aim: To construct the recombinant adenovirus expressing small hairpin RNA (shRNA) targeting human leukocyte-derived arginine aminopeptidase (LRAP) gene and silence the expression of LRAP in human retinal microvascular endothelial cells (HRMECs).

Methods: Three pairs of oligonucleotides coding for shRNAs targeting human LRAP gene were designed and synthesized, and were cloned into the shuttle vector pRNAT-H1.1/Adeno after annealing.

Biological safety of human skin-derived stem cells after long-term in vitro culture.

The aim of this study was to investigate the biological safety of human skin-derived mesenchymal stem cells (HSMSCs) cultured in vitro by detecting changes in karyotype, major histocompatibility complex expression and tumorigenicity. Before the 21st passage of the in vitro culture, cell surface markers were analysed by flow cytometry; major histocompatibility complex expression was detected by RT-PCR and flow cytometry. The tumorigenicity of HSMSCs was tested using SCID mice and observing changes in the injection site and pathological sections.