| In 2011, the US National Institute on Aging and the Alzheimer’s Disease Society(NIA-AA) released AD new clinical diagnostic guidelines for Alzheimer’s disease, dementia, dementia symptoms before the pre-clinical stage and asymptomatic phases are proposed the corresponding diagnostic criteria, emphasizing the importance of research that Aβ as biomarkers for diagnosis.But regardless of the cerebrospinal fluid or PET detect Aβ, is still carried out in the clinical routine difficulty.Looking for a new plasma-related biomarkers, especially the level of molecular biology markers provide the basis for early diagnosis and treatment of AD is a hot research.In recent years, Long noncoding RNA in the development of researchers revealed the importance of constantly being in the disease, more and more lnc RNAs associated with AD had found.By literature review, we found that BACE1-AS is one of the most important discoveries..As we all know, the abnormal deposition of Aβ is the key to the pathogenesis of AD.The Aβ is formed by the action of β- amyloid precursor protein(β-amyloid precursor protein, APP) in the β-secretase enzyme(β-site APP cleaving enzyme, BACE) and γ-secretase.BACE1 with it’s transcription of its antisense strand formed long noncoding RNA BACE1-AS form a duplex structure complex,to avoid the BACE1 m RNA from nuclease degradation, increase its stability, which leads to more Aβ deposition in brain tissue.However, the production of Aβ can be also used as a cytokine to promote the upregulation of BACE1-AS,enabling Aβ generated positive feedback regulation mechanism to accelerate the progress of the progress of AD.Studies have shown that in AD brain BACE1-AS transcripts of high specificity,but in the transcription of patient plasma have not been reported.In our study,we used guanidine isothiocyanate-phenol chloroform technology to extract BACE1-AS of Chinese Han AD patients’ plasma, and by sequencing analysis consistent with the reported sequence and analysis of the development of the role of the lnc RNA occurred in AD, for BACE1-AS as AD treatment target and plasma molecular markers to provide the research foundation.In the study,the subjects are collected from our hospital outpatient and inpatient treated possible clinical AD patients and medical center for sex,age and education-matched healthy human,a total of 55 cases.Participants signed informed consent, and blood samples were collected.Collected blood samples are centrifuged and the supernatant was saved, followed by acid guanidinium thiocyanate phenol chloroform method of plasma samples for RNA extraction, application software design primer Primer Premier5.0; reverse transcription of RNA extracted; the success of reverse transcriptase PCR amplification of a target gene and gel electrophoresis gene sequencing; imaging using image analysis software Chromas comparative analysis consistency sequencing sequence genebank sequences; analysis of target gene transcription differences between the two groups by the statistics.Result:Gene sequencing results by the National Center for Biotechnology Information(National Center of Biotechnology Information, NCBI) BLAST to compare, fully consistent with the confirmed target gene BACE1-AS.The sequencing results showed that the target gene was successfully extracted from the AD group in 5 cases, reaching 18.5%. Control group successfully extracted the target gene in 0 cases, accounting for 0%. There was significant difference between the two groups(P<0.05).Conclusion:The presence of AD in patients with plasma lnc RNA BACE1-AS, and lnc RNA can be stably present in the plasma. There were significant differences between the AD group and the control group, which were closely related to the AD and BACE1-AS, and the specificity was higher. Provide theoretical basis for BACE1-AS as a biomarker for the diagnosis of AD.The innovation of this research:For the first time transcription AD patient plasma long noncoding RNA BACE1-AS is detected and sequenced, which is not reported at home and abroad at present. |
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