Alzheimer disease diagnosis and treatment marker and application thereof

A kind of Alzheimer's disease, drug technology, applied in the treatment, human COPG1 gene in the field of diagnosis of Alzheimer's disease

Active Publication Date: 2015-10-14
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite the discovery of such a significant genetic risk associated with AD, as a complex disease, other risk genes must exist
[0003] At present, symptomatic treatment is mainly carried out in the early stage, and the clinically diagnosed AD patients are basically in the middle and late stage. The existing treatment can only improve the symptoms, but cannot prevent or reverse the progression of the disease.

Method used

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  • Alzheimer disease diagnosis and treatment marker and application thereof
  • Alzheimer disease diagnosis and treatment marker and application thereof
  • Alzheimer disease diagnosis and treatment marker and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 Screening for Gene Markers Related to Alzheimer's Disease

[0054] 1. Collection of peripheral blood samples

[0055] AD patients came from Beijing 301 Hospital, a total of 60 cases, aged 53-84 years old, all cases were diagnosed as AD, and its diagnostic criteria refer to the third revised edition of the American Diagnostic and Statistical Manual of Mental Disorders. A total of 50 cases were selected as the control group, selected from the routine physical examination population of Beijing Hospital. All the subjects were excluded from blood lipid metabolism and other diseases, aged 60-82 years. All subjects signed the informed consent for the testing project and provided peripheral blood for genetic testing.

[0056] 2. Total RNA extraction from blood

[0057] Using Biotek Blood RNA Extraction Reagent Kit for extraction of total RNA from blood

[0058] (1) Take 250μl (or 0.25g) of whole blood into the RNase-Free filter column, centrifuge at 13000rpm for ...

Embodiment 2

[0088] Example 2 COPG1 Gene Expression Plasmid Construction

[0089] 1. The human nerve cell line SH-SY5Y was incubated at 37° C. in 5% CO with DMEM (high glucose) medium containing 10% calf serum. 2 , Cultivated in an incubator with a relative humidity of 90%. The medium was changed once every 2-3 days, and 0.25% trypsin was used for routine digestion and passage.

[0090] 2. Overexpression of COPG1 gene

[0091] 2.1 Construction of COPG1 gene expression vector

[0092] Amplification primers were designed according to the coding sequence of the COPG1 gene (as shown in SEQ ID NO.1), and the primer sequences were as follows: the forward primer was 5'-TCGACAAGAAGGATGAGGAGT-3' (SEQ ID NO.7), and the reverse primer was 5' -TTATCCCCAGATGCCAAGA-3' (SEQ ID NO. 8). The coding sequence of the full-length COPG1 gene was amplified from the cDNA library of adult fetal brain (clontech company, catalog number: 638831), and the above cDNA sequence was double-digested with restriction end...

Embodiment 3

[0101] Example 3 Antagonistic effect of COPG1 gene expression on neuronal cell death induced by Aβ

[0102] 1. Cell transfection: The neural cell line R2L1 was transfected with pcDNA3.1-COPG1 and pcDNA3.1 according to the method in Example 2.

[0103] 2. After 24 hours of transfection, the neural cell line R2L1 was cultured in a 96-well plate with a cell density of 0.5×10 4 cells / well. Divide the cells into the following groups:

[0104] Uninduced group: transfected with pcDNA3.1, without adding 20 μM Aβ42;

[0105] Negative control group (pcDNA3.1+Aβ42): pcDNA3.1 was transfected, and 20 μM Aβ42 was added to the medium at the same time;

[0106] COPG1 gene overexpression group (pcDNA3.1-COPG1+Aβ42): transfect pcDNA3.1-COPG1, and add 20 μM Aβ42 to the medium at the same time;

[0107] Three replicate holes were set up for each group. After incubation at 37° C. for 20 h, MTT was added and incubated for 4 h. After adding the dissolving solution, incubate at 37°C for another...

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Abstract

The invention discloses an application of COPG1 gene in early diagnosis and treatment of Alzheimer disease, belonging to the technical field of new application of genes. Experiments prove that the expression of COPG1 gene in the blood of Alzheimer disease patients is reduced in comparison with normal people, indicating that the expression level of COPG1 gene in the blood of a subject can be measured to judge whether the subject suffers the Alzheimer disease. In the invention, A beta neurotoxicity experiments indicate that overexpression of COPG1 gene can effectively inhibit the neurotoxicity of A beta mediation. Therefore, the COPG1 gene can be used for preparing a medicine for treating or preventing Alzheimer disease and has a good prospect in development and application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of human COPG1 gene in the diagnosis and treatment of Alzheimer's disease. Background technique [0002] Alzheimer's disease (AD) is the most common progressive neurodegenerative disease with cognitive, behavioral and functional disturbances and is highly heritable (up to 76% heritability), but From a scientific point of view, the mechanism is particularly complex. As the population ages, the risk of developing dementia after the age of 85 can reach 50%, and dementia will also become a major challenge for human medicine. In the early genetic studies of AD, three genes involved in early-onset AD, late-onset AD and autosomal dominant genetic diseases were successfully identified, and showed complex inheritance patterns. The development of AD is due to the interaction between environmental risk factors and genes, and association studies have identified rare variants that cause...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68A61K48/00A61K38/17A61K45/00A61P25/28
CPCA61K38/1709A61K45/00A61K48/005C12Q1/6883C12Q2600/158G01N33/6896G01N2333/47G01N2800/2821
Inventor 杨承刚肖枫
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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