1.Effect and mechanism of cardiosphere-derived cells in the treatment of heart failure with preserved ejection fraction in rats
Xuefeng TIAN ; Guijia ZHU ; Sining LIU ; Ning LU ; Yue ZHANG ; Zheng WANG
Chinese Journal of Primary Medicine and Pharmacy 2017;24(16):2401-2405,后插1
Objective To test whether cardiosphere-derived cells(CDCs)were sufficient to decrease manifestations of heart failure with preserved ejection fraction(HFpEF)in hypertensive rats.Methods DS rats(Charles River,Wilmington,Massachusetts)were fed 0.3% NaCl(low-salt)diet until 7 weeks of age.At that time,the diet was switched to an 8% NaCl(high-salt)diet in rats by random assignment.DS rats fed the low-salt diet constituted the control group(n=20).At 13 to 14 weeks of age,rats with the high-salt diet were randomized to receive allogeneic rat CDCs or PBS.Echocardiography long-axis images of the left ventricular systolic and diastolic dimensions.Sirius red was used to assess fibrosis and proliferation.Results E/A ratio increased in the PBS-treated group compared with the control group and the CDCs-treated group [(1.20±0.30)vs.(1.70±0.20)or(1.80±0.16),t=0.782,0.844,all P<0.001].LAA kept increasing in the PBS-treated group[(27.20±1.10)mm2 vs.(19.80±0.76)mm2 or(17.80±0.82)mm2,t=0.892,0.774,all P<0.001].The time constant of isovolumic LV pressure fall was prolonged in placebo-treated animals compared with CDCs-treated animals and control rats[(6.2±0.3)×103 mmHg/s vs.(9.4±0.4)×103 mmHg/s,t=0.382,P=0.024;(6.2±0.3)×103 mmHg/s vs.(9.1±0.5)×103 mmHg/s,t=0.883,P=0.022].LVEDP was 2-fold higher in placebo-treated group than in CDC-treated and control animals[(17±10)mmHg vs.(8±3)mmHg,t=0.922,P=0.003;(17±10)mmHg vs.(9±4)mmHg,t=0.922,P=0.004].A dramatic improvement of survival was observed in CDC-treated rats(Kaplan-Meier survival curves)(P=0.027).Cardiac myofibroblasts increased dramatically in PBS-treated(110/field vs.46/field,P<0.001).Inflammatory cytokines expression siginficantly increased in PBS-treated group.Conclusion CDCs improves survival in a rat model of HFpEF through reducing inflammation and fibrosis.
2.Identification of a pathogenic Aeromonas veronii isolated from rock carp, Procypris rabaudi
Chengke ZHU ; Guijia LIU ; Zhengshi ZHANG ; Decheng PU ; Long ZHU ; Chaowei ZHOU ; Luo LEI ; Zonglin ZHENG
Chinese Journal of Zoonoses 2017;33(6):526-534
In June 2016,a disease among the cultured rock carp (Procypris rabaudi) in Yongchuan of Chongqing Municipality occurred.The aim of this study was to investigate biological characteristics and provide reference for Aeromonas veronii identification diagnosis and treatment.Pathogenic bacteria strain YY01 from the dying fishes were examined and isolated.Strain YY01's taxonomic status was identified by observing the morphology,studying the physiological and biochemical characters and sequencing the 16S rRNA and housekeeping gene gyrB.Its pathogenicity was checked by artificial infection experiment and virulence genes.Furthermore,effective medicine was detected by drug sensitivity.The 16S rRNA and gyrB gene sequence of the strain YY01 was more than 99% homology with that of Aeromonas veronii,suggesting that the pathogen was Aeromonas veronii,which was also identified by the results of biochemical analysis.The LD50 of strain YY01 to rcok carp was 5.06 × 104 CFU/g.Four virulence genes were detected from YY01,including aerolysin (aer),hemolysin (Hly),Outer Membrane Protein Gene A (OmpA) and adhesion (Aha) genes.Antibiotic sensitivity assays showed that among 40 antibiotics tested,22 were sensitive and 11 were resistant.In conclusion,the strain YY01 is identified as Aeromonas veronii and it is proved to have strong pathogenicity.
3.Study of fractional flow reserve guided percutaneous coronary intervention on non-culprit moderate coronary stenosis
Guijia ZHU ; Yangyang LIU ; Yang CAO ; Lin CHEN ; Duirong SUN ; Guo DONG ; Runtao GAN ; Shusen YANG
Chinese Journal of Interventional Cardiology 2014;(10):626-631
Objective To study the feasibility of using fractional flow reserve (FFR) to guide whether to perform coronary revascularization of non-culprit moderate stenosis in patients with unstable angina and estimate their clinical prognosis. Methods This study enrolled unstable angina patients with multivessel disease. First successful stenting of the culprit artery, then the other non-culprit moderate coronary stenosis were randomized into PCI guided by angiography or guided by FFR measurements. Death from any cause, nonfatal myocardial infarction, unplanned hospitalization leading to urgent revascularization and clinical manifestations with angina were followed during the first year. Results 71 patients were included, among them 35 patiens were randomly assigned to angiography-guided PCI and 36 patients to FFR-guided PCI. In FFR-guided PCI group, FFR was successfully measured in all of non-culprit moderate coronary stenosis. In 23 stenosis, the FFR was greater than 0.80, and stents were not placed in these stenosis. In 13 stenosis with FFR<0.8, stent were inplant and FFR was raised≥0.95 after stenting. The percentage of patients who had a primary end-point event was higher in the angiography-guided PCI group than the FFR-guided PCI group (P<0.05). Neither the rate of mortelity from any cause nor the rate of non-fatal myocardial infarction had significant difference between the 2 groups. Related to the target vessels rates of nonfatal myocardial infarction (5.6%vs. 28.6%) and target lesion revascularization (5.6%vs. 31.4%) were statistically different (P<0.01 and P<0.05, respectively). Conclusions In patients with unstable angina, it is safe to use FFR values to guide decisions on the revascularization of angiographically moderate non-culprit stenosis. Routine measurement of FFR in addition to angiographic guidance, as compared with PCI guided by angiography alone, results in a significant reduction in major adverse events at 1 year, particularly in urgent revascularization, and clinical manifestations with angina get better.
4.Effect and mechanism of cardiosphere-derived cells in the treatment of rats with acute myocardial infarction ;after reperfusion
Xuefeng TIAN ; Guijia ZHU ; Sining LIU ; Ning LU ; Yue ZHANG ; Zheng WANG
Chinese Journal of Primary Medicine and Pharmacy 2017;24(2):165-168
Objective To investigate whether cardiosphere-derived cells (CDCs)can protect ischemia-reperfusion in acute myocardial infarction,and to explore its mechanism.Methods 7 -10 week-old female Wistar-Kyoto (WKY)rats were used for all in vivo experiment.Ischemia was induced for 45 min to allow reperfusion. Twenty minutes (or two hours)later,CDCs (or PBS control)were injected into the LV cavity with an aortic cross-clamp.After 48 hours and 2 weeks,representative echocardiography long-axis images of the left ventricular (LV) systolic and diastolic dimensions,the protein level of activated caspase-3 were observed,the apoptosis rate of myo-cardial cells and the infarct area of the heart were determined in those groups.Results Rats underwent 45 minutes of ischemia,followed by either 20 minutes or 120 minutes (delayed injection)of reperfusion prior to infusion of CDCs (cells per 100μL)or PBS control (100μL)into the LV cavity during aortic cross-clamp.Ejection fraction,as meas-ured by echocardiography,was significantly preserved in CDCs-treated animals at 48 hours with a 20 -minute,but not a 120-minute,delay of infusion(28.0% vs 38.0%,χ2 =7.340,P=0.008).CDCs-treated animals reduced percentage of infarct mass(6.2% vs 13.4%,χ2 =4.226,P=0.002;6.2% vs 13.5%,χ2 =1.853,P=0.003), infarct mass(6.2% vs 13.4%,χ2 =2.220,P=0.002;6.2% vs 13.5%,χ2 =3.119,P=0.003)treated with PBS control.CDC-treated animals reduced infarct size,relative to those of animals treated with PBS control(45.0% vs 24.0%,χ2 =4.825,P=0.008),less thinning of the LV anterior wall(1.96mm vs 1.45mm,t=0.897,P=0.028). Protein expressions of MMP-8 and CXGL7 were elevated in the infarct zone of hearts treated with CDCs(MMP-8:0.74 vs 0.56,t=0.657,P=0.014;CXGL7:0.44 vs 0.81,t=0.791,P=0.010).Conclusion CDCs is suggested to be a promising cell source to repair acute myocardial infarction through inhibiting apoptosis and reduce proinflam-matory cytokine expression.
5.Up-regulation of p21 activated kinase 4 expression in the retina of diabetes mice and its effects on the behavior and mitochondrial function in retinal vascular endothelial cells
Mingfei JIAO ; Hui LI ; Jingjing CAO ; Zhenyu KOU ; Guijia WU ; Aihua LIU ; Lijie DONG
Chinese Journal of Ocular Fundus Diseases 2023;39(5):401-407
Objective:To observe the effects of p21 activated kinase 4 (PAK4) on the mitochondrial function and biological behavior in retinal vascular endothelial cells.Methods:The experimental study was divided into two parts: in vivo animal experiment and in vitro cell experiment. In vivo animal experiments: 12 healthy C57BL/6J male mice were randomly divided into normal control group and diabetes group, with 6 mice in each group. Diabetes mice were induced by streptozotocin to establish diabetes model. Eight weeks after modeling, quantitative real-time polymerase chain reaction and Western blots were performed to detect the expression of PAK4 in diabetic retinas. In vitro cell experiments: the human retinal microvascular endothelial cells (hRMEC) were divided into three groups: conventional cultured cells group (N group), empty vector transfected (Vector group); pcDNA-PAK4 eukaryotic expression plasmid transfected group (PAK4 group). WB and qPCR were used to detect transfection efficiency, while scratching assay, cell scratch test was used to detect cell migration in hRMEC of each group. In vitro white blood cell adhesion experiment combined with 4 ', 6-diamino-2-phenylindole staining was used to detect the number of white blood cells adhering to hRMEC in each group. The Seahorse XFe96 cell energy metabolism analyzer measures intracellular mitochondrial basal respiration, adenosine triphosphate (ATP) production, maximum respiration, and reserve respiration capacity. The t-test was used for comparison between the two groups. Single factor analysis of variance was used for comparison among the three groups. Results:In vivo animal experiments: compared with normal control group, the relative expression levels of PAK4 mRNA and protein in retina of diabetic mice were significantly increased, with statistical significance ( t=25.372, 22.419, 25.372; P<0.05). In vitro cell experiment: compared with the N group and Vector group, the PAK4 protein, mRNA relative expression and cell mobility in the hRMEC of PAK4 group were significantly increased, with statistical significance ( F=36.821, 38.692, 29.421; P<0.05). Flow cytometry showed that the adhesion number of leukocytes on hRMEC in PAK4 group was significantly increased, and the difference was statistically significant ( F=39.649, P<0.01). Mitochondrial pressure measurement results showed that the capacity of mitochondrial basic respiration, ATP production, maximum respiration and reserve respiration in hRMEC in PAK4 group was significantly decreased, with statistical significance ( F=27.472, 22.315, 31.147, 27.472; P<0.05). Conclusion:Over-expression of PAK4 impairs mitochondrial function and significantly promotes leukocyte adhesion and migration in retinal vascular endothelial cells.