ABSTRACT
Objective:To explore the potential targets and pathways of steroid alkaloids<italic> </italic>from<italic> Solanum</italic> <italic>nigrum</italic> （SASN） in the treatment of non-small cell lung cancer （NSCLC） and analyze the possible mechanism. Method:The active SASN against NSCLC were searched from literature. Then potential targets of SASN were screened through SwissTargetPrediction and PharmMapper， and those of NSCLC through GeneCards. Venny was employed to yield the common targets of the two， and Cytoscape to construct the 'medicinal-component-disease-target' network. Metascape was applied to enrich the Gene Ontology （GO） terms and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathways of the common targets， and STRING was used to generate the protein-protein interaction （PPI） network， followed by screening of key targets by Cytoscape. Finally， Western blot was used to verify the effects of the medicinal on key targets. Result:A total of 6 active SASN were screened out： solasonine， solamargine， solasodine， solanocapsine， solanidine， and <italic>N</italic>-methylsolasodine， which had 96 potential anti-NSCLC targets. These targets mainly involved the pathways in cancer， proteoglycans in cancer， and Forkhead box protein O （FoxO） pathway. PPI network analysis demonstrated 15 key anti-NSCLC targets of SASN， such as mitogen-activated protein kinase （MAPK）1， MAPK8， MAPK14， protein kinase B （Akt1）， signal transducer and activator of transcription 3 （STAT3）， and proto-oncogene tyrosine protein kinase （SRC）. Meanwhile， Western blot results showed that SASN could significantly down-regulate the expression of the key proteins Akt1， SRC， and STAT3. Conclusion:We predicted the potential targets and pathways of SASN against NSCLC and obtained 15 key targets， from which we selected three key proteins for validation. The validation results were consistent with the prediction results. This paper is expected to lay a scientific basis for the subsequent in-depth study of the mechanisms of SASN against NSCLC.
ABSTRACT
[Objective]To establish a reliable and accurate preimplantation genetic diagnosis (PGD)method using multiple dis?placement amplification (MDA), which can be applied to the diagnosis of X-linked severe combined immunodeficiency disease (X-SCID).[Methods]Haplotype analysis for the X-SCID family was performedusing five short tandem repeats (STR) markers flanking the both sides of the interleukin-2 (IL-2) receptor gamma chain (IL2RG) gene. MDA technique was used for single-cell whole genomic amplification. The products were used as template in polymerase chain reaction (PCR) of informative STR markers found by linkage analysis for haplotype analysis as well as sequencing of the IL2RG gene exon 5.The amelogenin (AMEL) locus was used to do sex diag?nosis.[Results]Linked analysis revealed 3 STR markers were informative. The method was evaluated with 10 single lymphocytes and 10 single blastomeres. MDA was successful in all single cell. The detection efficiency of gene sequencing of pathogenic IL 2RG exon5 was 100%. The PCR efficiency of 3 STR informative markers and AMEL was 96.3%(77/80)and the average allele drop-out (ADO) rate was 11.5%(7/61). A cycle of PGD was performed on the family, and seven embryos were diagnosed, two of which were normal embry?os. Twin pregnancy occurred after transplantation which were given a healthy baby boy and a healthy baby girl.[Conclusion]In this study, multiple displacement amplification combined with specific amplification/sequencing of pathogenic gene and haplotype analysis in the single cell level of X-linked severe combined immunodeficiency disease were performed. The protocol can avoid misdiagnosis caused by contamination and ADO, and improve the diagnostic efficiency of PGD.