1.Peroxisome proliferator-activated receptor γ nuclear translocation induced by focal cerebral ischemia-reperfusion in rats
Li SUN ; Yanwei XU ; Hao LIANG ; Guomin SUN ; Yan CHENG
Chinese Journal of Neurology 2010;43(7):512-515
objective To examine nuclear transIocation of peroxisome proliferator-activated receptor γ(PPARγ)in rats following focal cerebral ischemia/reperfusion(I/R),and to explore the significance of altered PPARγ,nuclear translocation in ischemic brain injury.Methods Healthy adult male SD rats underwent 60-min cerebral artery occlusion followed by reperfusion of 4,8,or 24 h,respectively.The cytoplasmic-to-nuclear shuttling of PPARγ was characterized by Western blot,immunohistochemical and immunofluoreseence staining.The effects of PPARγ agonist rosiglitazone (Ros) and antagonist GW9662 on I/R-induced PPARγ nuclear translocation were also examined in the present study. Furthermore,TTC staining war adopted to determine the change in cerebral infarction volume. Results (1)Western blot analysis revealed an increase of PPARγ in the nucleus and a simultaneous reduction in the cytosol following ischemia and reperfusion for 4 h(tcytosol=9.03,tmuclear=27.19,P=0.00).Prolonged the reperfusion further enhanced this I/R induced PPARγ translocation in a time-dependent manner.Using immunohistochemistry and immunofluorescence,nuclear PPAR γ positive staining increased from 48.3%in the sham control to 80.3% following ischemia and reperfusion for 24 h.(2)Western blot analysis revealed that PPARγ agonist Ros further increased I/R-induced nuclear enrichment of PPARγ,whereas PPARγ antagonist GW9662inhibited I/R-stimulated change in PPARγ.(3)When compared to the L/R group using TTC staining,Ros treatment significantly decreased the infarction volume by 48.40%(15.46±4.94 versus 29.96±3.39,t=5.93.P=0.00),whereas GW9662 increased by 58.95%(47.62±4.93 versus 29.96±3.39,t=7.23,P=0.00).Conclusions Cerebral I/R injury induces PPARγ translocation from the cytosol to the nucleus.This change may represent an intrinsic neuroprotective response against brain I/R injury.
3.The protective function of peroxisome proliferator-activated receptor gamma agonist to focal cerebral ischemia-reperfusion injury in mice model
Jinlin SHANG ; Li SUN ; Hao LIANG ; Yan CHENG
Chinese Journal of Neurology 2009;42(3):190-194
Objective To investigate neuroprotective function of peroxisome proliferator-activated receptor gamma (PPARγ) agonist, rosiglitazone against reperfusion injury after focal cerebral ischemia in mice model.Methods To establish cerebral isebemia-reperfusion injury mice model, adult male mice underwent 2 hours of middle cerebral artery occlusion followed by 22 hours reperfusion (MCAO/R). One hour before MCAO/R, mice were treated with either vehicle (MCAO/R group) or rosiglitazone (6 mg/kg, rosiglitazone group). 2,3,5-triphenyhetrazolium chloride (TIC) staining was applied to determine the volume of cerebralinfarction.TheneurologicaldeficitwasscoredatZeaLonga 5-pointscale. Myeloperoxidase (MPO) activity was measured in brain tissue as an index of neutrophil accumulation. RT-PCR, immunohistochemistry and Western blot were performed to examine the mRNA and protein expression of pro-inflammatory mediators (ICAM-1, IL-1β and COX-2).Results (1) The volume of cerebral infarction in rosiglitazone group was significantly decreased from that of MCAO/R group ( 29. 1 ± 6. 6 vs 57.8 ± 9. 7 ,t = 5. 980, P < 0. 01 ), and rosiglitazone markedly improved neurological function in treated mice than MCAO/R mice(1.2 +0.4 vs 3.3 ±0.8, t =5.812, P<0.01). (2) Compared with MCAO/R group, MPO activity in the rosiglitazone-treated group was significantly lower ((0. 049 + 0. 005 ) U/g vs (0. 083 ±0. 008) U/g,t =5. 904, P <0. 01 ). (3) The mRNA and protein expression of pro-inflammatory mediators (ICAM-1, IL-1β and COX-2) in rosiglitazone group were also significantly decreased from those in MCAO/R group, as demonstrated by RT-PCR (0.313 ±0.024, 0.205 ±0.007, 0.359 ±0.060, t = 7.464, 19.656, 29.319, P <0.01, respectively) and Western blot (0.274±0.014, 0.205±0.025, 0. 146±0.015, t=79.909, 21.392, 95. 105, P<0.01, respectively). ConclusionThe present study suggests that PPARγ agonist, rosiglitazone, has neureprotective properties to cerebral ischemia-reperfusian injury and that the protection is partially mediated via anti-inflarmmatory actions.
4.BPD surgery on type 2 diabetes mellitus in GK rats
Liang CHENG ; Xuejun SUN ; Shiyun ZHANG ; Qilong WANG
Journal of Endocrine Surgery 2012;6(6):371-374
Objective To investigate the therapeutic effects and mechanism of biliopancreatic diversion (BPD) surgery on type 2 diabetes mellitus(T2DM) in GK rats.Methods 16 GK rats were randomly divided into 2 groups:BPD surgery group included 10 rats undergoing BPD surgery,sham-BPD group included 6 rats undergoing a sham operation.Fasting plasma glucose,insulin,glucagon-like peptide-1 (GLP-1) and glucose-dependent insulinotropic polypeptide(GIP)were detected one week before BPD surgery and the 1st week,4th week,10th week,26th week after BPD surgery.Oral glucose tolerance test(OGTT) and insulin tolerance test(ITT) were done in the 10th week after BPD surgery.Results There was no statistical difference in fasting plasma glucose,insulin,plasma GLP-1 or GIP between the 2 groups before surgery.Plasma glucose had significant reduction in BPD group compared to that in the sham group(P <0.05) and insulin level had no significant difference between the 2 groups.Rats in BPD group had significant improvement in glucose tolerance and insulin sensitivity compared to those in the sham group.Serum level of GLP-1 was significantly elevated in BPD group compared to that before surgery (P =0.0337 at the 1st week after surgery; P =0.0002 at the 4th week after surgery,P < 0.0001 at the 10th week after surgery,P <0.0001 at the 26th week after surgery) and that in sham-BPD group(P =0.0354 at the 1st week after surgery,P =0.0032 at the 4th week after surgery,P =0.0001 at the 10th week after surgery,P <0.0001 at the 26th week).Serum level of GIP was significantly lowered in BPD group compared to that before surgery(P =0.0189 at the 1st week after surgery; P =0.0007 at the 4th week after surgery,P =0.0003 at the 10th week after surgery,P <0.0001 at the 26th week after surgery) and that in sham-BPD group(P =0.0089 at the 1st week after surgery,P =0.0002 at the 4th week after surgery,P =0.0006 at the 10th week after surgery,P <0.0001 at the 26th week after surgery).The difference had statistical significance (P <0.05).Conclusion BPD surgery can significantly reduce fasting plasma glucose,improve glucose tolerance and insulin sensitivity.The change of serum levels of GLP-1 and GIP may play the major role in BPD treatment of diabetes mellitus.
5.Clinical significance of BP1 gene expression in human thyroid cancer
Qilong WANG ; Xuejun SUN ; Lingu WANG ; Shiyun ZHANG ; Liang CHENG
Journal of Endocrine Surgery 2013;7(3):218-220
Objective To detect the expression of BP1 gene in thyroid cancer and its relationship with clinicopathological features of thyroid cancer.Methods BP1 gene expression in 60 cases of thyroid cancer tissues and 20 cases of normal thyroid tissues were detected by in situ hybridization and immunohistochemistry.Results The positive expression rate of BP1 mRNA was 78.3 % (47/60) in the 60 cases of thyroid cancer tissues while it was 20% (4/20) in the 20 cases of normal thyroid tissues detected by in situ hybridization.The difference had statistical significance (P < 0.05).Of the 3 pathological types of thyroid cancer,the positive expression rate of papillary carcinoma was 81.6% (40/49),follicular carcinoma 85.7% (6/7),and medullary carcinoma 25.0% (1/4).The expression of BP1 mRNA had statistical difference between medullary carcinoma and other pathological types like papillary carcinoma and follicular carcinoma (P < 0.05).The positive expression rate of BP1 protein was 93.3% (56/60)in the 60 cases of thyroid cancer tissues while it was 10.0% (2/20) in the 20 cases of normal thyroid tissues detected by immunohistochemistry.The difference had statistical significance(P <0.05).Conclusion BP1 gene expression is up-regulated in human thyroid cancer and it is related to tumor stage and pathological type but not related to patients' age,sex or lymph node metastasis.
6.The inhibitory effect of PNS on brain 3-NT formation in vitro
Chen XIAO ; Li SUN ; Shanshan CAO ; Hao LIANG ; Yan CHENG
Tianjin Medical Journal 2017;45(3):229-233
Objective To study the inhibitory effect of panax notoginseng saponins (PNS) on 3-nitrotyrosine (3-NT) formation in brain induced by heme/NO2 -/H2O2 or ONOO - pathways in vitro. Methods According to the two major pathways of 3-NT formation in vivo, the models of protein nitration induced by heme/NaNO2/H2O2 or ONOO-system were established, respectively, in vitro. Bovine serum albumin (BSA)/rat plasma protein or rat brain homogenate protein were utilized as reactive substrates in both systems. Samples were divided into blank-control group, 3-NT group and PNS group (including low-, medium-and high-concentration subgroups). In 3-NT group, samples were exposed to heme/NaNO2/H2O2 or ONOO-system, respectively, at 37℃for 30 min, whereas in PNS group, samples were pre-incubated with PNS (at final concentrations of 50 mg/L, 100 mg/L, and 200 mg/L) at 37℃for 5 min before the nitrating system exposure. The 3-NT level in each group was detected by Western blot assy. Results Compared with the blank-control group, both heme/NaNO2/H2O2 and ONOO-system can induce significant 3-NT generation in BSA/rat plasma protein or rat brain homogenate protein (P<0.05). Compared with model group, PNS pre-treatment markedly inhibited 3-NT expression in BSA/rat plasma protein in a dose-dependent manner (P<0.05), the inhibitory effect of low intervention on the level of 3-NT in rat brain homogenate protein was not significant (P>0.05). Medium- and high-concentrations of PNS pre-treatment markedly inhibited 3-NT accumulation, with maximum effect at the concentration of 200 mg/L (P<0.05). Conclusion Medium- and high-concentrations of PNS can inhibit 3-NT formation in brain tissue mediated by either heme/NO2-/H2O2 or ONOO-pathways, implying that potential neuroprotective action against 3-NT involves pathological conditions, like trauma, stroke, and neurodegenerative diseases.
7.Tongxinluo enhancing proliferation of peripheral blood-derived human endothelial progenitor cells
Xiao-Wei LIANG ; Cheng-Bo SUN ; Hua WANG ; Chun LIANG ; Zong-Gui WU ;
Academic Journal of Second Military Medical University 1985;0(06):-
Objective:To study the effects of Tongxinluo,a traditional Chinese medicine,on the proliferation of peripheral blood-derived endothelial progenitor cells(EPCs)in vitro.Methods:The Tongxinluo solution was prepared through ultrasonication according to the pervious literature.The mononuclear cells(MNCs)were isolated from the peripheral blood with Ficoll by density gradient centrifugation.MNCs were suspended in Medium 199 containing 20% fetal blood serum(FBS)and vascular endothelial growth factor(VEGF).After cultured for 7 d,the attached cells were characterized by Di-LDL uptaking and FITC-lectin binding by laser confocal microscope,and further identified through detection of CD34 and CD133 expression by flow cytometry.Then the cultured EPCs were incubated with Tongxinluo at a series of concentrations(0,50,100,200,500, 750,1000?g/ml)for different durations(0,6,12,24 and 36 h).The cell morphology was observed and cell proliferation was determined by MTT assay.Results:Incubation with different concentrations of Tongxinluo increased the proliferative ability of EPCs.Tongxinluo at 500?g/ml had the most prominent effect on proliferation and the effect increased as time went by and reached peak at 36 h(growth rate 54.18%,P
9.Risk factors of postoperative death in patients with intraventricular hemorrhage
Yanfei ZHANG ; Zhongwei ZHUANG ; Weifeng ZHANG ; Cheng YANG ; Liang WEI ; Zhiyang SUN
Chinese Journal of Emergency Medicine 2012;21(5):527-530
Objective To study the risky factors related to postoperative mortality after surgical intervention for intra-ventricular hemorrhage (IVH).Methods The clinical data of 142 IVH patients aged from 19 to 86 years with 77 male and 65 female were collected from 2007 to 2010 for retrospective analysis with SPSS version 13.0 package.Multi-factorial Logistic regression analysis and mono-factorial x2 analysis were used to find out the risk factors related to post-operative death. Results Of 142 patients,48 (33.8%) died.Mono-factorial analysis demonstrated that the risk factors of death after surgical intervention to IVH were elderly age ( > 65 years),history of hypertension,low GGS score,intra-ventricular diffuse bleeding,preoperative obstructive hydrocephalus and coagulation abnormality,postoperative central high pyrexia and central diabetes insipidus,postoperative hemorrhage of alimentary tract,pulmonary infection,and drainage after operation ≥7 days ( P < 0.05 or P < 0.01 ).Multi-factorial Logistic regression analysis,on the other hand,revealed the independent factors associated with postoperative IVH mortality were postoperative central high fever,preoperative obstructive hydrocephalus (P < 0.01 ); drainage after operation ≥7 days,postoperative pulmonary infection,elderly age,low GCS score,intra-ventricular diffuse bleeding and history of hypertension ( P < 0.05 ). Conclusions General condition of patients with IVH and CT findings are likely related to mortality,and active prevention against various complications is the crucial approach to lowering postoperative mortality and improving prognosis.
10.Comparison of temperature curve and ablation zone between 915 MHz and 2450 MHz cooled-shaft microwave antenna:results in ex vivo porcine livers
Yuanyuan SUN ; Ping LIANG ; Lei DONG ; Zhigang CHENG ; Qiujiu SHAO ; Yang WANG ; Yongyan GAO
Chinese Journal of Ultrasonography 2011;20(11):992-995
Objective To compare temperature curve and ablation zone between 915 MHz and 2450 MHz cooled shaft microwave antenna in ex vivo porcine livers.Methods The 915 MHz and 2450 MHz microwave ablation and thermal monitor system were used in this study.A total of 56 ablation zones and 280 temperature data were obtained in ex vivo porcine livers.The output powers were 50,60,70,and 80W and the setting time was 600s.The temperature curve of every temperature spot,the short- and long-axis diameters of the coagulation zones were recorded and measured.Results At all four power output settings,the peak temperatures of every temperature spot had a tendency to increase accordingly as the output power was increased,and except for 5 mm away from the antenna,the peak temperatures for the 915 MHz cooledshaft antenna were significantly higher than those for the 2450 MHz cooled-shaft antenna (P <0.05).Meanwhile,the short- and long-axis diameters for the 915 MHz cooled-shaft antenna were significantly larger than those for the 2450 MHz cooled-shaft antenna ( P <0.05).Conclusions The 915 MHz cooledshaft antenna can yield a significantly larger ablation zone and achieve higher temperature in ablation zone than 2450 MHz cooled-shaft antenna in ex vivo porcine livers.