1.Screening of LRRK2 interactants by yeast 2-hybrid analysis
Xiaoye ZHENG ; Mei YANG ; Jieqiong TAN ; Qian PAN ; Zhigao LONG ; Heping DAI ; Kun XIA ; Jiahui XIA ; Zhuohua ZHANG
Journal of Central South University(Medical Sciences) 2008;33(10):883-891
Objective To isolate and identify the potential binding partners of LRRK2,a gene linked to both dominant familial form and sporadic form of Parkinson's disease,thus to further our knowledge of its function.Methods We used a sequence containing full-length of COR domain and part of ROC and MAPKKK domain as bait.The bait amplified by polymerase chain reaction(PCR) was then cloned into a yeast expression plasmid pGBKT7.After being sequenced and analyzed,pGBKT7-bait was transformed into the yeast strain AH109.Western blot was performed to confirm the expression of pGBKT7-bait in AH109 yeast strain.Then human fetal brain cDNA library was trarnsformed into that yeast strain.which could express pGBKT7-bait fusion protein.The yeast strain which contained pGBKT7-bait and human fetal brain cDNA library was plated on quadruple dropout medium (SD/-Trp/-Leu/-His/-Ade)containing X-a-gal.We retested these positive colonies using 2 independent yeast strains AH109 contained pGBKT7-bait or pGBKT7,respectively.At last,these plasmids isolated from these true positive colonies were analyzed by bioinformatics.Results We obtained 9 true positive colonies,these colonies were sequenced, and we performed sequence Blast in GenBank.Three colonies of the 9 positive colonies were not in open reading-frames.Among other 6 colonies,there were known proteins including spermatid perinuclear RNA-binding protein(STRBP)and BCL2-associated athanogene 5 isoform b(BAG5),as well as unknown proteins including tyrosine phosphatase non-receptor type(PTPN23),1(3)mbt-like 3 isoform b(L3 MBTL3),RALY RNA binding protein-like isoform 1(RALYL),and Homo sapiens mRNA for KIAA1783 protein,partial cds(KIAA 1783).Conclusion True positive colonies of LRRK2 are successfully obtained by the yeast 2-hybrid.Our screened proteins may provide a new research clue for revealing biological functions of LRRK2,pathogenesis of Parkinson's disease,and other neurodegerations.
2.Combination of low-dose HBIg and Nucleoside analogues to prevent recurrent hepatitis B virus after liver transplantation: a retrospective analysis of 1506 cases
Zhongyang SHEN ; Zhijun ZHU ; Yonglin DENG ; Liying SUN ; Wei QU ; Wei RAO ; Xiaoye SUN ; Hong ZHENG ; Cheng PAN ; Yihe LIU
Chinese Journal of Hepatobiliary Surgery 2011;17(5):364-366
Objective To evaluate the preventive effect of combination of low-dose HBIg and Nucleoside analogues on recurrence of hepatitis B after liver transplantation. Methods Retrospectively analyzed HBV status and recurrence in patients accepting Nucleoside analogues plus low-dose HBIg as prophylaxis treatment after liver transplantation for HBV-related end-stage liver disease from December 1998 to Octomber 2009 in our center. Results In all the 1506 patients whose survival time >30 d after liver transplantation, 37 patients showed HBV recurrence, the HBV cumulative-recurrence rate of 1, 2, 3, 4, 5 and 6y was 1.3%,2. 4%,2. 7%,2. 9%,3. 7% and 4.6% respectively. The time of recurrence varied from 0. 3 to 66. 6 months (median 12. 8 months) after transplantation. Virus mutation could be tested in 9 cases of the 37 recurrence patients, including 4 YMDD cases, 2 YMDD + YIDD cases, 1 YMDD+YVDD cases, 1 YVDD case,and 1 YIDD case. Conclusions Liver transplantation is the principal therapeutic method for the patient with end-stage liver diseases related to HBV, with the effectively prophylaxis treatment to aim directly at HBV recurrence. If the patients who got HBV recurrence received targeted treatments, the situation can be controlled satisfactorily.
3.Relationship between self-management ability and self-efficacy in patients with coronary heart disease in a community of Shanghai
Chen CHEN ; Derong PENG ; Fenhong YANG ; Xiaoye PAN ; Weigang XU
Chinese Journal of General Practitioners 2020;19(11):1020-1024
Objective:To investigate the relationship between self-management ability and self-efficacy in coronary heart disease (CHD) patients, and the influential factors.Methods:From April to June 2016, a questionnaire survey was conducted among 236 patients with coronary heart disease selected by simple random sampling method in the outpatient clinic of Pengpu Community Health Service Center; and the Coronary Artery Disease Self-management Scale and self-efficacy questionnaire were used in the survey. The correlation between scores of the two scales was analyzed; the scores were compared between patients with percutaneous coronary intervention (PCI) and those without PCI, between patients with myocardial infarction (MI) and those without MI. Multiple linear regression was performed to elucidate the influencing factors of self-management ability and self-efficacy of CHD patients.Results:The mean scores of self-management ability and self-efficacy were (75.5±15.5) and (24.8±10.8), respectively. The self-management ability was significantly correlated with self-efficacy in CHD patients(r=0.398-0.683, P<0.05). The self-management score of patients with PCI was higher than those without PCI[ (80.4±14.9) vs.(72.5±15.1), t=3.38, P<0.01]. The scores of self-management[(85.0±16.4) vs.(72.9±14.2), t=5.14, P<0.01]and self-efficacy[(29.3±12.0) vs. (23.6±10.2), t=3.39, P<0.01] of patients with MI were higher than those without MI. Multiple linear stepwise regression analysis indicated that age over 65, history of MI, history of PCI, history of smoking and self-efficacy score were all factors influencing self-management behavior in patients with CHD ( R=0.735; R2=0.540; F=26.451, P<0.01). Smoking history and self-management score were the influencing factors of self-efficacy in patients with CHD ( R=0.690; R2=0.476; F=20.425, P<0.01). Conclusion:The self-efficacy of patients with CHD is closely related to their self-management behavior, so we should strengthen their self-efficacy and improve their self-management level by comprehensive measures, particularly for patients without PCI.