1.Research progress on animal models of Parkinson's disease
Zhicheng ZHANG ; Yuan YUAN ; Xuan WANG ; Qingkai SONG ; Jiejie DAI
Chinese Journal of Comparative Medicine 2018;28(5):21-27
Parkinson's disease(PD)is a progressive neurodegenerative disorder, with an etiology that is now considered to be due to interaction between genetic and environmental factors. Typical PD features include loss of dopaminergic neurons in the nigrostriatal region, with typical motor traits of PD associated with dopamine deficiency. Animal models have contributed to determining PD etiology and pathogenesis,as well as testing new therapeutic schedules and novel drug research. Rodents, tree shrews, primates, and other animal models of PD have been established by different method. These models each have their own advantages and limitations, showing different clinical features and pathological mechanisms to those in humans. Therefore, the appropriate model for scientific research must be carefully considered. This article reviews the main neurotoxic and transgenic models of PD.
2.Cloning and Bioinformatics Analysis of GI4CL Gene in Glehnia littoralis
Jiejie SONG ; Hongmei LUO ; Xunzhi ZHU ; Yu ZHANG ; Ting GAO
World Science and Technology-Modernization of Traditional Chinese Medicine 2017;19(4):610-617
This study was aimed to clone and analyze the open reading frame (ORF) of 4-coumarate:coenzyme A ligase (Gl4CL) gene in Glehnia littoralis.Based on the high-throughput sequencing of G.littoralis,the full-length cDNA of Gl4CL gene was cloned by the rapid amplification of cDNA ends (RACE) method.Physical and chemical properties,secondary structure and three-dimensional structure of Gl4CL protein were predicted.Real-time PCR was used to detect the expression of Gl4CL gene in roots and leaves of G.littoralis.A total of 1951 bp full-length cDNA of Gl4CL gene was obtained,which encoded a protein of 544 amino acids with a predicted molecular weight of 59.481 kDa and the isoelectric point of 8.20.The cDNA of Gl4CL gene included 1 635 bp of ORF,153 bp of 5'untranslated regions (5'UTR) and 163 bp of 3'UTR.The result of real-time PCR showed that Gl4CL gene was both expressed in roots and leaves of G.littoralis,while the expression of gene in roots was significantly higher than that in leaves.It was concluded that the study will lay the foundation for further study of Gl4CL gene in function and gene regulation.Through in-depth study of the relationship between the expression of Gl4CL gene and lignin,as well as the plant growth phenotypes,it is expected to obtain high yield and quality lines of Glehniae Radix with strong resistance to diseases and insect pests.
3.Establishment and application of a TaqMan real-time fluorescence quantitative PCR for detection of tree shrew adenovirus(TAV)
Qingkai SONG ; Xiaofei LI ; Yurun MIAO ; Zhicheng ZHANG ; Xuan WANG ; Yuan YUAN ; Jiejie DAI ; Xiaomei SUN
Chinese Journal of Comparative Medicine 2018;28(3):72-77
Objective To establish a quick and accurate method for detection of tree shrew adenovirus(TAV) using TaqMan real-time fluorescence quantitative PCR. Methods Based on the published TAV genome sequence, a 3' conserved sequence was used to design specific probe primers. A standard curve was prepared using a recombinant plasmid containing the target gene fragment. A real-time fluorescence quantitative PCR method was established for detecting TAV based on TaqMan probe. Results The detection method was specific and was not cross-reactive with other common pathogens. The detection limit of the method was 3.7 copies/μL,showing a high sensitivity. The correlation coefficient was 0.998, and the efficiency was 95.7%. The amplification result showed a fine linear relationship,and the repeatability test effect was good. Conclusions The TAV real-time quantitative PCR detection method based on TaqMan probe has been successfully established. It has high sensitivity and reproducibility and can be applied to early detection of TAV infection.
4.Mechanism of Astragalus methylside alleviating cardiomyocyte hypertrophy
Jiejie ZHANG ; Xiqing WEI ; Bingchun SONG ; Youqian LI ; Hongsheng ZHANG ; Zhencai GAO ; Cheng SHEN ; Jinguo ZHANG
Chinese Journal of Geriatrics 2022;41(7):837-842
Objective:To investigate the protective effects and related mechanisms of Astragaloside Ⅳ(ASⅣ)alleviating Angiotensin II-induced cardiomyocyte hypertrophy.Methods:H9c2 cardiomyocytes were divided into six groups: normal control group, ASⅣ group(ASⅣ 100 μmol/L), AngⅡ group(AngⅡ 1 μmol/L), and three ASⅣ dose experiments(AngⅡ 1 μmol/L + ASⅣ 25 μmol/l group, AngⅡ 1 μmol/L+ ASⅣ 50 μmol/l group, AngⅡ1 μmol/L+ ASⅣ 100 μmol/L group), and simultaneously cultured for 24 hours.Cardiomyocyte viability was assessed by CCK8 assay, and surface area of culturedcardiomyocytes in each group was assessed by immunofluorescence assay.Atrial natriuretic peptide(ANP)mRNA expression was assessed by fluorescence real-time quantitative RT-PCR.And LC3 protein expression, an autophagy related protein, was assessed by Western blotting as well as immunofluorescence.Results:(1)AngⅡ decreased cardiomyocyte H9c2 viability in a dose-dependent manner( P<0.05). ASⅣ could inhibit the decrease of cardiomyocyte H9c2 viability in response to AngⅡ in a dose-dependent manner( P<0.05). (2)H9c2 cardiomyocytes induced by AngⅡ showed a significantly larger cell area and significantly higher ANP mRNA and ANP protein expression compared with controls.Different concentrations of ASⅣ intervention could reverse the increase of cardiomyocyte H9c2 area induced by AngⅡ and also decreased the expression of ANP protein induced by AngⅡ in a dose-dependent manner(all P<0.05). (3)Compared with the control group, the autophagy level and the expression of autophagy marker LC3II/I of H9c2 cardiomyocytes induced by AngⅡ were significantly increased(all P<0.05). ASⅣ could inhibit AngⅡ-activated autophagy, and the difference was statistically significant( P<0.05). ASⅣ inhibited the expression of LC3II/I in H9c2 cardiomyocytes stimulated by AngⅡ, and the difference was statistically significant( P<0.05). Conclusions:ASⅣ inhibits AngⅡ-induced cardiac hypertrophy by inhibiting autophagy of cardiomyocytes.