[0062] Example 1
[0063] 1.1 Research object
[0064] We selected 80 Chinese Han nationality patients with a clear diagnosis of coronary heart disease. They all took nitroglycerin as medicine for angina pectoris. According to the effect of sublingual nitroglycerin administration, these patients can be divided into an effective group that is effective within ten minutes, and an ineffective group that is not effective within ten minutes. There are 59 patients in the effective group, including 47 males and 12 females. There were 21 patients in the ineffective group, including 13 males and 8 females. In these patients, from the perspective of gender relations, the effect of drug administration is not significantly related to the gender of the patient (p=0.107).
[0065] 1.2 Experimental methods and results
[0066] 1.2.1 DNA extraction
[0067] DNA was extracted from human blood by conventional phenol-chloroform method, and the concentration was adjusted to 20ng/ul, and then used for conventional PCR amplification.
[0068] 1.2.2 Design of PCR and sequencing primers
[0069] According to the ALDH2 gene sequence in GenBank, the following primers were designed and synthesized. The specific primers are shown in Table 1 below.
[0070] Primer name
[0071] 1.2.3 PCR amplification of ALDH2 gene
[0072] Using the extracted DNA as a template, Taq enzyme was used to perform PCR amplification on the GeneAmp 9700 PCR machine with the Touchdown program. The reaction conditions are: pre-denaturation at 94°C for 2 minutes, denaturation at 94°C for 30 seconds, annealing at 63°C for 40 seconds, and extension at 72°C for 40 seconds, a total of 10 cycles, with each cycle annealing temperature decreasing by 0.5°C; subsequent denaturation at 94°C for 30 seconds, Annealed at 58°C for 40 seconds, and extended at 72°C for 40 seconds, a total of 30 cycles; finally, it was extended at 72°C for 7 minutes. The PCR products were verified by agarose gel electrophoresis.
[0073] As a result, an amplified product of 358 bp was obtained.
[0074] 1.2.4 SNP discovery and detection
[0075] After the PCR product is purified by Resin resin, use ABI-PRISM TM 377 DNA sequencer (applied biosystems (ABI)) was used for fluorescent labeling end-termination of two-way sequencing, using Polyphred software (University of Washington http://droog.mbt.washington.edu/Polyphred.html) Interpretation and SNP confirmation.
[0076] As a result, the following SNP was found: 1951 G→A.
[0077]1.2.5 SNP genotyping and analysis
[0078] These 80 patients were genotyped for the 1951G/A site in the acetaldehyde dehydrogenase 2 gene using the above PCR and sequencing results.
[0079] The results showed that: in the effective group, the number of homozygous individuals of acetaldehyde dehydrogenase 2 genotype 1 (ALDH2*1) (that is, G/G at position 1951) was 40; while in the ineffective group, the number of acetaldehyde dehydrogenase The number of homozygous individuals with 2 genotype 1 (ALDH2*1) is 7. The number of homozygotes for acetaldehyde dehydrogenase 2 gene type 1 (ALDH2*1) was significantly different in the two different groups (x2=7.59, p=0.006). The results show that the effect of sublingual nitroglycerin administration in the treatment of acute angina pectoris is significantly greater for individuals who are homozygous for acetaldehyde dehydrogenase 2 gene type 1 (ALDH2*1) than for acetaldehyde dehydrogenase 2 gene type 2 (ALDH2*1). 2) Individuals who are homozygous (that is, A/A at position 1951), or heterozygous for acetaldehyde dehydrogenase 2 genotype 1 and type 2 (that is, G/A at position 1951) are more effective, and both have significant Sexual difference.