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Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient

A technology of methamphetamine and factor, applied in the field of protein detection

Active Publication Date: 2012-10-03
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the relationship between complement factor H (Complement factor H, CFH) and METH

Method used

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  • Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient
  • Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient
  • Application of complement factor H (CFH) to genetic expression products of methamphetamine (METH) addiction patient

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1 Preparation of human serum samples and isolation of CFH

[0074] 1. Subject population

[0075] METH-addicted patients were recruited from Shenzhen Compulsory Isolation Drug Rehabilitation Center with the assistance of China Institute of Drug Dependence, Peking University; normal controls (healthy volunteers) were recruited from Peking University Third Hospital. After all subjects signed the informed consent, they were evaluated and diagnosed by addiction-related professionals. The selection of patients with METH addiction is based on the "Measures for the Identification of Drug Addiction" of the People's Republic of China (Decree No. 115 of the Ministry of Public Security), "Guiding Principles for the Diagnosis and Treatment of Amphetamine Dependence" and relevant international standards, the tenth edition of the International Classification of Diseases (International Classification of Diseases, ICD-10) and amphetamines in the fourth edition of the Diagnos...

Embodiment 2

[0105] Example 2 Conditioned place preference (CPP) experiment in rats and the acquisition of rat serum and related brain tissue

[0106] In order to confirm the results of serum proteomics in patients with METH addiction, we prepared a rat model of METH addiction, and after the successful verification of the model by CPP experiment, we obtained the model's serum and addiction-related brain tissue for verification of differentially expressed proteins.

[0107] Male SD rats, 30, were then divided into a control group (group C) and a model group (group M), 15 in each group. "Handle" the rat every day (catch the rat with the glove used for drug administration and stroke its abdomen) to adapt to the subsequent intraperitoneal drug administration, and start the formal experiment after 3 days of adaptation. The formal experiment is divided into three parts: pre-conditioning test, conditioning and post-conditioning test (see Figure 17 A). The main experimental methods are as fol...

Embodiment 3

[0116] Example 3 Western blot analysis

[0117] In order to verify the results obtained by 2-DE and mass spectrometry, and to confirm the differential expression of CFH in the serum of addicted people and rats and normal control serum, the following Western blot analysis experiments were performed:

[0118] ① Western blot analysis of patient serum

[0119] Serum samples were diluted 30-fold with Tris-HCl (0.01M, pH 7.4) buffer and loaded. SDS-PAGE used 7.5% polyacrylamide gel as the separating gel. 5% skimmed milk was used for blocking, the primary antibody was CFH monoclonal antibody (Santa Cruz, U.S.A), and the dilution ratio was 1:500. The secondary antibody was HRP-labeled IgG (Proteintech Group, U.S.A), with a dilution ratio of 1:2500. To ensure consistent protein loading, an internal reference was added. GAPDH was used as an internal reference, the primary antibody was GAPDH polyclonal antibody (Millipore, U.S.A), and the dilution ratio was 1:1000; the secondary ant...

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Abstract

The invention relates to a detection method for methamphetamine (METH) addiction and application of a complement factor H (CFH) to genetic expression products of an METH addiction related group. On the basis of a great number of experiments, the stable expression of the CFH, alpha-1-acid glycoprotein 2, transthyretin, apolipoprotein L1 and haptoglobin in the blood serum of an METH addiction patient is up-regulated, and the CFH, the alpha-1-acid glycoprotein 2, the transthyretin, the apolipoprotein L1 and the haptoglobin can serve as the genetic expression products which are related to METH addiction. The content of CFH proteins in human serum is quantitatively detected by using an immunoblotting method or an enzyme linked immunosorbent assay kit, and expression up-regulation of the content of the CFH proteins is applicable to a method for identifying the METH addiction.

Description

technical field [0001] The invention relates to a protein detection method, in particular to a detection method for methamphetamine addiction and the application of complement factor H (Complement factor, CFH) as a gene expression product of methamphetamine (METH) addiction-related populations. Background technique [0002] Drug abuse and drug dependence are not only common public health problems in the world, but also serious social problems. Drug dependence is a mental, and sometimes physical, state resulting from the interaction of a drug with the body, manifesting a compulsive need for continuous or periodic use of the drug, behaviors and other responses in order to experience it Psychological effects, or to avoid discomfort caused by drug withdrawal, including both mental and physical dependence. Methamphetamine (Methamphetamine, METH), commonly known as methamphetamine, belongs to amphetamine-type stimulants (amphetamine-type stimulants, ATS), its structure is similar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N27/447G01N1/28
Inventor 赵欣史琬璐蒲小平刘志民
Owner PEKING UNIV
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