Objective To evaluate the effect of celastrol postconditioning on focal cerebral ischemiareperfusion (I/R) injury in rats.Methods Sixty-four Sprague-Dawle rats (32 males,32 females),weighing 250300 g,were randomized into 4 groups using a random number table (n =16 each):sham operation group (group S) ; celastrol control group (group S + C) ; focal cerebral I/R group (group I/R) ; celastrol postconditioning group (group I/R + C).Focal cerebral I/R were produced by middle cerebral artery occlusion (MCAO).Dimethyl sulfoxide (DMSO) 0.3 ml/kg was injected intraperitoneally after shame operation in group S.Celastrol 3 mg/kg was injected intraperitoneally after shame operation in group S + C.DMSO 0.3 ml/kg was injected intraperitoneally at 5 min of reperfusion in group I/R.Celastrol 3 mg/kg was injected intraperitoneally at 5 min of reperfusion in group I/R + C.The neurologic deficit was scored at 5 min before reperfusion and 24 h of reperfusion.The infarct size was detected by TTC staining,and then the percentage of infarct size was calculated.The pathological changes in CA1 region of ischemic hippocampus were detected by HE staining.The activity of nicotinamide adenine dinucleotide phosphate oxidase (NOX),content of reactive oxygen species (ROS) and expression of NOX1 and NOX2 mRNA (by RT-PCR) in ischemic brain tissues were detected.Results Compared with S and S + C groups,the neurologic deficit scores,infarct size,percentage of infarct size,NOX activity and ROS content were significantly increased,and the expression of NOX1 mRNA and NOX2 mRNA was up-regulated in I/R and I/R + C groups (P ＜ 0.01).Compared with group I/R,the neurologic deficit scores,infarct size,percentage of infarct size,NOX activity and ROS content were significantly decreased,and the expression of NOX1 mRNA and NOX2 mRNA was down-regulated in group I/R+ C (P ＜ 0.01).There was no significant difference in the parameters mentioned above between group S and group S + C (P ＞ 0.05).The pathological changes in CAl region of ischemic hippocampus were significantly attenuated in group I/R + C (P ＜ 0.01).Conclusion Postconditioning with celastrol can auenuate focal cerebral [/R injury in rats and inhibiton of oxidative stress response in brain tissues may be involved in the mechanism.

Endothelial progenitor cells have self-replication and multi-differentiation potentials. They participate in the maintenance of vascular dynamics and physiological reconstruction. Though the research of endothelial progenitor cells has become a current hot spot, there are a series of unknown factors from the fields of the basic research to the clinical application. This article reviews the advances in research on endothelial progenitor cells,particularly the roles in ischemic cerebrovascular disease.

[Objective] To explore the counteractive mechanism of Tongmai Injection (IT) for excitotoxicity in rats with cerebral ischemia/reperfusion by observing the glutamic acid (Glu) content and N-methyl-D-aspartate (NMDA) receptor activity in rats cortex. [ Methods ] Rat models with cerebral ischemia/reperfusion were established by four-vessel occlusion. Glu content and NMDA receptor activity were examined by radioimmunoassay and the effects of TI on Glu and NMDA receptor were also observed. [ Results ] Glu content and NMDA receptor activity were both increased in the model group and TI could counteract the above changes. [Conclusion] Cerebral ischemia/reperfusion can induce excitotoxicity and TI can protect the cerebral cortex.

Objective To investigate the effects of EGB on hepatic fibrosis and expression of Activin A in rats with fibrosis.Methods From Oct.2004 to Apr.2005,the study was conducted in 30 adult male rats in the pepartment of Gastroenterology,the people's Hospital of Wuhan University.The rats were randomized into 3 groups:control group,model group and treatment group.Except the rats in the control group,others were induced to hepatic fibrosis by intraperitoneal injection of CCl_4 twice a week for 8 weeks.Those rats in the treatment group were intragastrically administered with EGB establish model of everyday.At the end of the 8th week,all rats were sacrificed.The samples of liver was staining with HE.The expression of Activin A was determined by immunohistochemistry and RT-PCR.Results The grade of fibrosis in EGB-treated groups were lower than that in the model group(P

Objective To evaluate the role of angiotensin-converting enzyme (ACE) gene polymorphism in the progression of renal diseases. Methods ACE gene polymorphism was analyzed in 77 patients with end stage renal failure (ESRF) and 150 normal control. Results The frequences of DD and DI genotype were significantly higher in ESRF patients than normal control (DD, 15.6% VS 6.0%, P

Objective A LC/MS/MS method is developed for determination of paraquat in biological tissue.Methods The samples were pretreated with solid phase extraction using Oasis WCX cartridges and separated with HPLC,paraquat could be identified by LC/MS/MS.Result Calibration curves were linear on injection of amounts ranging from 0.02～20?g/ml and the limit of detection was 10ng/ml(S/N≥3).Conclusion The described menthod was proved to be sensitive,rapid and accurate,it will be applied in identification and determination of paraquat in biological tissues.

AIM:Activin A,a member of transforming growth factor superfamily,is the negative regulator factor in liver regeneration. In this study,the effects of extract of Ginkgo Biloba on hepatic fibrosis and the expression of Activin A in rats with cirrhosis were investigated. METHODS:The experiment was performed at in the Central Laboratory of Wuhan First Hospital from September 2005 to December 2006. ①Thirty-six male SD rats of(160?20) g were randomized into 3 groups:control group,model group and treatment group. ② Except the rats in the control group,others were intraperitoneally injected with 500 mL/L CCl4 for 8 weeks to establish models of hepatic fibrosis. Meanwhile,the extract treatment group was infused with the extract of Ginkgo Biloba(Chinese drugs preparation laboratory of Wuhan First Hospital,detected by Hubeu Wushi Medicine Industry Co.,Ltd. No. 02-391) daily for 8 weeks. ③After administration,all anesthetized rats were sacrificed. Blood samples were collected for the determination of liver function biochemical indexes. Liver tissue samples were used for histopathological examinations. The expression of Activin A was determined by immunohistochemistry and RT-PCR. RESULTS:All 36 rats were involved in the final analysis. ①The liver function in extract treatment group was significantly improved compared with that in model group. ②The grade of fibrosis in extract treatment group were remarkably lower than that in model group under light microscope. ③The positive staining of Activin A in treatment group was significantly reduced compared with model group. ④The expression of Activin A mRNA in extract treatment group was significantly reduced compared with model group. CONCLUSION:Extract of Ginkgo Biloba can effectively decrease the expression of Activin A in rats with hepatic fibrosis caused by CCl4,and lessen the degree of hepatic fibrosis.

Objective To investigate the effect of autologous bone marrow-derived endothelial progenitor cell (EPC) transplantation on neurological outcomes in cerebral ischernia in rats and its poss le mechanisms.Methods Autologous bone marrow-derived EPC was cultured in vitro and it was labeled with 5-bromodeoxyuridine (BrdU).A middle cerebral artery occlusion (MCAO) model was induced by the intraluminal suture method.The rats in a EPC group transplanted autologous EPC (106/ml/kg) via external jugular veins,those in a control group were injected with phosphate buffered saline (1 ml/kg),and those in a sham operation group (n =15)were not treated.The modified neurological severity score (mNSS) was used to observe the neurological changes of the rats.BrdU immunohistochemical staining was used to evaluate EPC proliferation and differentiation.Three-dimensional confocal image analysis was used to detect the vascular structure and density in cerebral ischemic areas.TUNEL staining was used to detect the apoptotio cells in ischernic brain tissue.Enzyme-linked immunosorbent assay was used to detect the concentration of plasma vascular endothelial growth factor VEGF).Results The mNSS in the EPC group was siginficantly lower than that in the control group (at day 8:6.43 ±0.69 vs.8.86 ±0.95,q =2.673,P=0.035; at day 14:4.55 ±0.89 vs.6.73 ± 1.06,q =5.360,P =0.035).The number of BrdU positive cells in the EPC group was significantly higher than that in the control group (42.2±5.76 vs.25.67±5.49,q=4.020,P=0.030).The capiilary diameter in the EPCgroup was significantly smaller than that in the control group (4.51 ± 0.21 μm vs.6.34 ± 0.24 μm,q =3.980,P =0.003); the density of blood vessels (212.64 ± 8.02/0.002 mm3 vs.153.60 ± 7.21/0.002 mm3; q =9.670,P =0.001 ) and the total surface area of microvessel (92 013 ± 5 132 μm2/0.002 mm3 vs. 71 366 ±4 538 μm2/0.002 mm3; q=4.180,P=0.014) were significantly higher or more than those in the control group.The number of apoptotic cells in the EPC group was significantly less than that in the control group (36.26 ± 6.91 vs.78.34 ± 7.21; t =-4.834,P =0.003).The plasma VEGF concentration in the EPC group was significantly higher than that in the control group (54.91 ± 5.71 pg/ml vs.13.81 ± 4.25 pg/ml,q =12.300,P=0.002).Conclusions Autologous EPC transplantation has a protective effect on ischemic brain tissue in rats.It may be associated with VEGF related angiogenesis and neuroprotection.It has an important application prospect in the treatment of ischemic cerebrovascular disease.

Objective To investigate the neuroprotective effect of escitalopram on focal cerebral ischemia/reperfusion in rats and its possible mechanisms.Methods Seventy-five male Sprague-Dawley rats were randomly divided into three groups:sham operation,saline control and escitalopram intervention groups (n =25 in each group).A focal cerebral ischemia reperfusion model in rats was induced by the intraluminal suture method.The modified neurological severity scale was used to evaluate neurological deficit in rats (n =5 in each group).Laser confocal technology was used to observe the microvascular diameter,density,and total area in ischemic region (n =5 in each group).Enzyme-linked immunosorbent assay was used to detect the plasma concentration of vascular endothelial growth factor (VEGF) (n =5 in each group).Immunohistochemical staining (n =5 in each group) and Western blotting (n =5 in each group) were used to detect the expression of VEGF in the ischemic brain tissue.Results At day 14 after modeling,the neurological deficit improved more significantly in the escitalopram intervention group than that in the saline control group (4.39 ±0.92 vs.6.57 ± 1.13; P =0.015).The 3D confocal vascular imaging showed that capillary diameter in the escitalopram intervention group was significantly smaller than that in the saline control group (2.93 ± 0.19 μm vs.3.56 ± 0.22 μm; P ＜0.01); the vascular density was significantly higher than that in the saline control group (232.68 ±12.54/0.002 mm3 vs.176.26 ± 10.87/0.002 mm3; P=0.000); the total microvascular area was significantly greater than that in the saline control group (89 154± 3 298 μm2/0.002 mm3 vs.75 368.14± 3 519 μm2/0.002 mm3; P=0.000).Enzyme-linked immunosorbent assay showed that the plasma VEGF concentration in the escitalopram intervention group was significantly higher than that in the saline control group (50.35 ± 5.44 pg/ml vs.13.75 ± 4.12 pg/ml; P =0.000).Immunohistochemical analysis showed that the VEGF expression in ischemic brain tissue in the escitalopram intervention group was significantly higher than that in the saline control group (P =0.000).Western blotting showed that the VEGF expression in ischemic brain tissue in the escitalopram intervention group was significantly higher than that in the saline control group (0.94 ±0.18 vs.0.62 ±0.22; P =0.006).Conclusions Escitalopram may reduce neurological deficit in cerebral ischemia/reperfusion in rats.Its mechanisms may be associated with VEGF-mediated angiogenesis.

Objective To explore the effects of granulocyte colony stimulating factor (G-CSF)on chronic cerebral ischemia in rats,and its possible mechanism.Methods Chronic cerebral ischemia (2-VO) model was prepared and bilateral external jugular veins were isolated.A total of 30 rats were divided into 2 groups at random sham group (received no intervention,n=15) and operative group (received G-CSF or PBS through external jugular vein injection,n=15).At 6 weeks after operation,the rats in operative group were divided into G--CSF group (received G-CSF 10 mg/L,1 ml · kg-1 · d-1,1 times every 24 h for,3 times) and PBS control group (received PBS 10 mg/L,1 ml ·kg 1 · d-11,1 times every 24 h for 3 times).At 8 weeks after the operation,morris water maze was carried out to evaluate the learning and memory ability of the rats.The cell proliferation,threedimensional vascular distribution,ischemic neuronal apoptosis,cell morphological changes in ischemic area and the plasma VEGF levels were detected to explore the possible mechanisms.Results In morris water maze,escape latency at the 2rd to 5th day were significantly lower in G-CSF group than the PBS group (all P＜0.05).The swimming time spent in the first quadrant in G-CSF group was significantly longer than the PBS group (P＜0.05).There was a significant difference in the number of BrdU positive cells in the ischemical area between the G-CSF group and the control group [(27.7±4.76) vs.(10.4 ± 3.7),P =0.030).Three-dimensional quantitative measurements of vascular structure showed that the capillary diameters was smaller in the G-CSF group than in the PBS group [(2.90±0.20) μm vs.(3.45±0.26) μm,P=0.020] and the number of branch points in the boundary regions of ischemia had a significant difference in the G-CSF group compared with the control group [(207.82±10.73) /0.002 mm3 vs.(162.10±9.31) /0.002mm3,P=0.005].Threedimensional cerebral vessel surface area in the ipsilateral hemisphere was increased in the G-CSF group compared with the PBS group [(86498±2896) μm2/0.002 mm3vs.(73976±3826) μm2/0.002 mm3,P=0.003].The number of apoptotic cells in G-CSF group was decreased compared with the PBS group [(32.10±6.70) vs.(56.30±11.20),F=11.89,P=0.043].The electron microscope morphological observations showed inflammatory edema in intercellular gap was significantly reduced in the G-CSF group compared with the PBS group.The level of plasma VEGF was significantly increased in the G-CSF group compared with the PBS group [(58.81±6.61) ng/L vs.(20.81±4.35)ng/L,P=0.025].Conclusions G--CSF can improve the learning and memory ability in the chronic cerebral ischemic rats,and its possible mechanism might involve the nerve protection and the vascular regeneration associated with the VEGF.There is a great prospect for G-CSF in the therapy of chronic cerebral ischemic disease.