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Objective @#The CRISPR / Cas9 technology was applied to construct PDE4D homozygous knockout mice to provide a basis for in-depth investigation of PDE4D gene function and mechanism of action．@*Methods@#A vector was constructed for PDE4D gene exon 4，5 microinjected into fertilized eggs of C57BL /6J mice，and PDE4D －/ - mice were obtained after maternal breeding and offspring mating，and the mice genotypes were determined by PCR product sequencing and genotype identification techniques．Changes in morphology and function of the major organs of the mice were detected using an ultrasound imaging system and H＆E staining，and the expression of PDE4D protein in the mice was verified by Western blot assay. @*Results @#The PDE4D －/ － mouse genotype was stably inherited， the mice were small，and there were no obvious morphological and histological changes in the major organs in vivo. The PDE4D expression was reduced or largely absent in the major tissues of PDE4D heterozygous or pure knockout mice，and the knockout effect was better．@*Conclusion @#PDE4D －/ － mice were successfully established using CRISPR / Cas9 technology，and no significant physiological abnormalities were found，which could be used for disease pathogenesis and drug research using PDE4D as the target.
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Objective To investigate the association between PDE4D gene rs918592 site polymorphisms and both vulnerable plaque and carotid intima-media thickness (CIMT) in patients with acute atherosclerotic cerebral infarction.Methods Five hundred and two patients with first onset of cerebral infarction,admitted to our hospital from February 2010 to November 2012,were chosen in our study; polymerase chain reaction-restriction enzyme fragment length polymorphism (PCR-RFLP) was adopted to analyze PDE4D gene rs918592 site polymorphisms in these patients.Ultrasound was used to measure CIMT and evaluate the carotid plaques (vulnerable plaque and stable plaque).Results The AA+AG genotype frequency in vulnenrable plaque group was obviously higher than that in stable plaque group (87.0％ vs.75.5％,P=0.006,OR=2.170,95％CI:1.238-3.802).And the A allele frequency in vulnerable plaque group was significantly higher than that in stable plaque group (61.3％ vs.53.4％,P=0.029,OR=1.385,95％CI:1.033-1.856).CIMTs ofGG genotype and AA+AG genotype were (1.17±0.20) mm and (1.19±0.18) mm,respectively (t=0.556,P=0.579).These risk factors,including smoking history (P=0.039,OR=1.753,95％CI:1.029-2.987),low-density lipoprotein cholesterol level (P=0.000,OR=2.537,95％CI:1.599-4.024),and AA+AG genotype in PDE4D gene rs918592 site (P=0.017,OR=2.037,95％CI:1.137-3.651),were finally incorporated into the models of stable plaque group and vulnerable plaque group by non-conditional Logistic regression analysis.Conclusion AA+AG genotype in PDE4D gene rs918592 site is one of potential risk factors contributing the formation of vulnerable plaque in patients with acute atherosclerotic cerebral infarction.The A allele might be a genetic susceptibility gene which leads to carotid plaque rupture.
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0.05) . Conclusions The A-allele in rs918592 may one of the risk factors in development of ICVD in the Han people in China. PDE4D gene may be not included glycometabolic mechanism to effect ICVD.