1.Intravenous transplantation of GDNF gene-modified CD34~+ cells from human umbilical cord blood for cerebral ischemie-reperfusion inury in rats
Chinese Journal of Organ Transplantation 2010;31(3):162-166
Objective To observe the neurologicaI functional recovery after intravenous transplantation of human umbilical cord blood(HUCB)CD34~+ cells transfected with glial cell-derived neurotrophic factor (GDNF) and green fluorescent protein (GFP) in SD rats with middle cerebral artery occlusion (MCAO).and tO investigate the survival,migration and neural differentiation of the graft cells.Methods (1)CD34~+ cells were isolated from HUCB using centrifuge combined with immune beads and then identified by flow cytometry,and transfected by the recombinant plasmid of GDNF.GFP or GFP plasmid by liposome method.(2) Sixty aduh male SD rats with MCAO were randomly divided into three groups (n=20 in each group):GDNF-GFP-CD34~+ cells group,in which the GDNF-GFP-CD34~+ cells were transplanted intravenously at the 24th h after the establishment ofmodels of MCAO;GFP-CD34~+ cells group:in which the GFP-CD34~+ cells were transplanted intravenously at the Same time;Normal saline group,in which normal saline was injected at the same time.Fifteen SD rats served as sham-operated group.(3) Neurological functional measurements were performed using the modified neurolc'gical severity score.Quantitative histological determinations of infarct volume were performed using standard TTC staining and quantitative image analysis. The GDNF level in the cell culture or the cerebral tissue was measured by ELISA. Meanwhile, the survival and migration of GFP-labeled CD34~+ cells and the expression of astrocytie marker-GFAP and the neuron marker-neuronal nuclei (NeuN) were detected by immunohistochemical and fluorescent staining. Results (1) The GDNF level in the cell culture was significantly higher in GDNF-GFP-CD34~+ cells than in GFP-CD34~+ cells (P<0. 05). (2) No significant difference was found in the modified neurological severity score at the day 7 after transplantation among the three groups with MACO, but at day the 28 after transplantation, the neurological function in GFP-GDNF-CD34~+ group was improved significantly (5.0±1. O) as compared with the GFP-CD34~+ cells (5. 9 + 1.4) or saline groups (7. 0±1.7) (P<0. 05). The cerebral infarct volume in GFP-GDNF-CD34~+ group (142±44mm~3) was significantly decreased as compared with the GFP-CD34~+ group (196±58 mm~3) (P<0. 05) or saline group (233<50 mm~3 ) (P<0. 01 ). The GDNF level in cerebral tissue in GFP-GDNF-CD34~+ group was significantly increased as compared with the GFP-CD34~+ group (P<0. 05) at the 28th day after treatment. At the 28th day after treatment, the NeuN positive cells (6.7±2.0), and GFAP positive ceils (14.1±3.3) in GFP-GDNF-CD34~+ group were significantly increased as compared with the GFP-CD34~+ group (P<0. 05), but there were no positive cells in sham-operationgroup. Conclusion Intravenous transplantation of GDNF gene-modified CD34~+ cells from human umbilical cord blood could improve the neurological function in rats with MCAO. The increased GDNF level in cerebral tissue was one of possible mechanisms responsible for the different improvements.
2.Effects of Lorsartan, Fosinopril on myocardial fibrosis, angiotensin Ⅱ and cardiac remolding in hypertensive rats
Bixiu HE ; Guolong YU ; Xiaoqiu LIANG
Journal of Central South University(Medical Sciences) 2001;26(2):118-120
Objective: To investigate effects of lorsartan, fosinopril on myocardial fibrosis, angiotensin Ⅱ and cardiac remolding in the spontaneously hypertensive rats (SHR). Methods: 16-week-old SHRs were divided randomly into 3 groups: SHR-L (treated with lorsartan), SHR-F (treated with fosinopril) and SHR-C (untreated), each group consisting of 10 rats. After 8 weeks' and 16 weeks' therapeutic period, collagen volume fraction (CVF), perivascular circuferential area (PVCA), plasma and myocardium angiotensin Ⅱ concentrations were examined by pathological examination with computed processing and radioimmunoassay respectively. Results: (1) Compared with SHR-C after 8 weeks' and 16 weeks' therapeutic period, the systolic blood pressure (SBP) was decreased similarly in both treatment groups. Heart and left ventricular weights, heart weight and eft ventricular mass indexes were lower significantly in both treatment groups than in SHR-C. Left ventricular mass index was reduced to a lower extent in SHR-F group than in SHR-L group after 16 weeks. (2) Compared with SHR-C, CVF, PVCA after 8 weeks and 16 weeks were reduced significantly in SHR-F and SHR-L. Meanwhile, CVF after 16 weeks in SHR-F than in SHR-L. (3) Compared with SHR-C after both therapeutic periods, plasma and myocardium angiotensin Ⅱ concentrations were increased Significantly in SHR-L, but plasma angiotensin Ⅱ concentrations were not altered significantly in SHR-F. However, myocardium angiotensin Ⅱ concentrations were reduced significantly in SHR-F after 8 weeks and 16 weeks in SHR-F. Conclusion: Lorsartan, fosinopril inhibit myocardial fibrosis and reverse heart hypertrophy. Fosinopril may be more effective in these above effects than Lorsartan. The mechanism of the both drug's cardioprotective effects was related to inhibition of myocardium rennin-angiotension-aldsteron system.
3.Life Quality and Related Factors of the Chronic Elderly Patients
Xiumei XIE ; Biefei LI ; Guolong YU
Chinese Mental Health Journal 1991;0(02):-
Objective: To study the quality of life of chronic elderly patients Method: 150 chronic patients aged 60 or above were assessed with SCL-90 (symptom checklist-90), SDSS (social deficit screening scale), ADL (activity of daily living) and QOL (quality of life), their demographic data were also collected Results: Men's social function was poorer than women's, but their quality of life had no significant difference to women's The quality of life in our sample differed with different age, educational degree, family conditions, occupations and incomes Conclusion: Life quality of chronic old patients is related to their demographic characters
4.Effects of high volume hemofiltration(HVHF) on cytokines in elderly patients with septic shock and MODS
Guolong CAI ; Jin YAN ; Yihua YU ; Zhaocai ZHANG ; Jiangou CHEN
Chinese Journal of Emergency Medicine 2006;0(01):-
Objective To study the effects of HVHF on cytokines in elderly patients with septic shock and MODS. Methods 22 elderly patients with septic shock and MODS were included in this prospective and interventional study. Before HVHF and 1,3,6,9,12,18 and 24 hour after HVHF, blood sample and ultrafiltration fluid were collected for cytokines analysis(radioimmunoassay). Results All patients finished HVHF. No severe side effects were observed,and the APACHEⅡ and MODS scores were decreased during HVHF(P
5.Experimental Animal Model of Focal Cerebral Ischemic-Reperfusion Injury in Spontaneous Hypertensive Rats
Guolong YU ; Ke HU ; Yali OU ; Tianlun YANG
Chinese Journal of Hypertension 2007;0(07):-
0.05).The cerebral infarcted area in SHR were significantly greater than that in SD rats [(42.6?5.6)% vs(29.5?6.7)%,P
6.Effects of intravenous transplantation of human umbilical cord blood mononuclear cells combined compound Danshen dripping pills on the microenvironment and apoptosis in the myocardium of the rabbits with acute myocardial infarction
Chunjun YUAN ; Qi AI ; Liuxia DENG ; Guolong YU
Journal of Central South University(Medical Sciences) 2013;38(8):785-792
Objective:To explore the effects of compound Danshen dripping pills (CDDP) and CDDP combined with transplantation of human umbilical cord blood cells (HUMNCs) on the inlfammatory response, oxidative stress, myocardial cell apoptosis and cardiac function, and also to investigate the possible mechanisms of the combined therapy in the acute myocardial infarction (AMI).
Methods:Rabbit model of AMI successfully established by ligation of the letf anterior coronary artery (LAD). Forty rabbits were randomly divided into 4 groups (n=10 per group):a control group, injected with 0.5 mL of saline in 24 h atfer AMI and then gavaged with 5 mL of saline daily;a CDDP group, injected with saline 0.5 mL atfer AMI and then gavaged with CDDP (270 mg/d) daily;a transplantation group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs [labeled with green fluorescent protein (GFP)] and then gavaged with 5 mL of saline daily;a combined group, injected with 0.5 mL of saline contained 3 × 107 HUCBMCs (labeled with GFP) and then gavaged with CDDP (270 mg/d) daily. Cardiac function index such as left ventricular fractional shorting (LVFS) and ejection fraction(LVEF) were measured by echocardiography;the pathological changes were observed by HE staining and the white blood cells in the myocardium were determined by light microscopy. hTe superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in myocardium were detected by nitrotetrazolium blue chloride (NBT) and thiobarbituric acid colorimetric measurement respectively. hTe number of transplanted cells in the myocardium was examined by GFP positive cells counted with lfuorescence microscopy.
Results:1) Compared with the control group (at 1 or 4 week), LVEF and LVFS were signiifcant improved in the CDDP group, the transplantation group and the combined groups (all P<0.05), the improvement degree of cardiac function in the combined group was the most significance. There was no significant difference between the CDDP group and the transplantation group. 2) Compared with the control group (at 1 or 4 week), the number of white blood cell, myocardial cell apoptosis ratio were decreased signiifcantly in the CDDP group, the transplantation group and the combined groups (all P<0.05), this decrease in the combined group was the most signiifcance, and there was no significant difference between the CDDP group and the transplantation group. 3) Compared with control (at 4 week), the SOD activity was increased signiifcantly, and MDA content in myocardium was decreased in the CDDP group, this change in the combined group was the most signiifcance. 4) GFP-positive cells were found to be present in the peri-myocardial infarction area in the transplantation group and the combined group at 1, 4 weeks post-transplantation. hTe number of the GFP positive cells in the combined group was more than that in the transplantation group (P<0.05).
Conclusion:The intravenous transplantation of HUMNCs combined with the CDDP in the treatment of rabbits with AMI could increase the survival rate of transplanted cells and inhibit the myocardial cell apoptosis, therefore improve the heart function. hTe possible mechanism of the combined treatment may be involved in the inhibition of the inlfammatory response and oxidative stress in the myocardium following AMI.
7.Influences of psycho-social factors in the treatment of the elderly patients with hypertension
Xiumei XIE ; Guolong YU ; Jin HE ; Biefei LI
Chinese Journal of Geriatrics 2003;0(07):-
Objective To study the influences of psycho-social factors in the treatment of the aged patients with hypertension. Methods Aged in-patients with hypertension were divided into two groups according to their improvement: 148 reached the standard and 62 did not reach the standard. Their psycho-social factors were assessed by self-rating depression scale(SDS), self-rating anxiety scale(SAS), life events scale (LES) and social support scale. Results (1)The percentages of depression (7.4% vs 32.2%, P
8.Effect of endothelial progenitor cells on the proliferation of co-cultured vascular smooth muscle cells
Li FANG ; Meifang CHEN ; Guolong YU ; Zhilin XIAO ; Xiaobin CHEN ; Xiumei XIE
Journal of Central South University(Medical Sciences) 2010;35(1):52-62
Objective To explore the effect of endothelial progenitor cells (EPCs) on the proliferation of co-cultured rat vascular smooth muscle cells(VSMCs). Methods Mononuclear cells were isolated from fresh cord blood by 6% hydroxyethyl starch (HES) and density gradient centrifugation. Isolated mononuclear cells were cultured in EGM-2 medium supplemented with 20% fetal bovine serum (FBS), VEGF,bFGF and other growth factors. Biological features of EPCs were observed at different time points, and EPCs were identified by morphology, fluorescence double-staining and flow cytometry. Indirect immunofluorescence was performed to analyze the expression of α-SM-actin, calponin of VSMCs special antigen. Co-culture system of EPCs and VSMCs was established by transwell permeable support. FBS (20%) was used to stimulate the proliferation of VSMCs. In a VSMCs/EPCs co-culture system, the DNA synthesis ability, total protein level and cell cycle of VSMCs were determined by BrdU marking method,protein quantitation and flow cytometry after co-culture for 6, 12, 24,48 and 72 h. Results After co-culture for 12, 24, 48, and 72 h, the DNA synthesis ability and total protein level of VSMCs significantly decreased compared with the control group(P<0.05). Flow cytometry showed that the percentage of S phase of VSMCs in VSMCs/EPCs co-cultured group significantly decreased and the percentage of G_1 phase increased markedly compared with the control group(P<0.05). The maximal inhibitory effect was observed at 48 h. Conclusion Early EPCs could inhibit the proliferation of VSMCs.
9.Role of antibody chin in analysis of inflammatory cytokine expression in severe sepsis
Jing YAN ; Zhaocai ZHANG ; Guolong CAI ; Yihua YU ; Caibao HU ; Liang WU ;
Chinese Journal of Emergency Medicine 2006;0(09):-
Objective To investigate the expression of inflammatory cytokines in patients with severe sepsis by using an antibody chip.Methods Twelve patients with severe sepsis and other 10 patients whose age and gender were matehed were enrolled in this study.Proteins from patients were labeled with biotin.The biotin-labeled proteins reaeted with antibody chips,on which there were antibodies of 40 major inflammatory cytokines.The target proteins were conjugated with streptomycin antibody labeled by horseradish peroxidase(HRP),and signals were imaged by laser scanner.Results In comparison with control group,the serum levels of inflammatory eytokines ineluding pro-and anti-inflammatory cytokines,ehemokines and certain eytokines receptors were notably increased,while expression of anti-inflammatory interleukin(IL)-2,-4,-13,-15 was remarkably decreased in sepsis group.Conclusions Excess inflammatory response and imbalance of pro-and anti-inflammatory eytokines were presented in the eourse of severe sepsis.
10.Assessment of fluid treatment in elderly patients with severe sepsis guided by monitoring the variation of stroke volume
Yihua YU ; Jing YAN ; Haiwen DAI ; Shijin GONG ; Guolong CAI ; Zhaocai ZHANG ; Jin CHEN
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
0.05).Conclusions Fluid therapy guided by functional hemodynamic index SVV in elderly patients with severe sepsis under MV is safe and more effective than that guided by CVP.SVV is valuable in prodicting the response to fluid treatment in elderly patients with severe sepsis.