1.Detection on early gene expression of tissues around hematom in rats with cerebral hemorrhage with the technique of DNA microarray
Chinese Journal of Tissue Engineering Research 2006;10(20):173-175
BACKGROUND: Changes of physiological structure, changes of phenotype and first basic excision are all the changes of gene expression. The technique of DNA microarray is a new method to filtrate target genes fleetly and largely by using the theory of base-partnershin, which can holistically and magnificently study the expression and function of organics genes. OBJECTIVE: To study early differential expression genes of rats with cerebral hemorrhage with DNA microarray and establish academic foundation for exploring mechanism of cerebral hemorrhage.DESIGN: Randomized controlled research.SETTING: Department of Neurology, Affiliated Hospital of North Sichuan Medical College. MATERIALS: The experiment was conducted at the Affiliated Hospital of North Sichuan Medical College from October 2002 to December 2003. Twenty Wistar rats, of either gender, with body mass of 220-260 g, without special pathogen, provided by Experimental Animal Center of Chongqing University of Medical Sciences, were selected and randomly divided into control group and cerebral hemorrhage group, with 10 in each group. METHODS: Animal models with cute cerebral hemorrhage of rats were established with type Ⅶ collagenase tridimensional localization method,and 4 hours later tissues around hematom and normal cerebral tissue at the same part were detected with gene chip. Fluorescent signal was scanned with scanning apparatus and analyzed with computer. Result of genic expressive pattern was researched with reverse transcriptase-polymerase chain reaction (RT-PCR). MAIN OUTCOME MEASURES: Result of gene chip in cerebral tissue of rats and result of RT-PCR.RESULTS: Four hours after acute cerebral hemorrhage, 129 differential expression genes were screened out, in which there were 114 up-regnlation genes and 15 down-regulation genes. Those genes were mostly related to the following aspects: stress, immunological response, apoptosis, energy metabolism and signal transmitting. Genes related with inflammatory impairment were mostly obvious. The result of RT-PCR suggested that the level of genic expression was as the same as the result of Cdna chip, which indicated that genic expressive pattern based on gene chip had great reliability.CONCLUSION: Early cerebral hemorrhage has many differential expression genes, which can play an important role in hemorrhagic brain damage.
2.Research on experimental intracerebral hematoma with haemadipsa yanyuanensis injection
Wenbin WU ; Changlin HU ; Bihua WU
Journal of Clinical Neurology 1997;0(06):-
Objective To investigate the effect of purified haemadipsa yanyuanensis injection on promoting intracerebral hematoma in rats and its possible mechanism.Methods We set up the experimental intracerebral hemorrhage (ICH) model in rats by stereotaxically injecting quantitative collagenase into their left caudate nucleuses.The effects of haemadipsa yanyuannesis injection on hematoma volume,neurological function recovery,brain water content(BWC),as well as histopathological changes were observed. BWC was calculated by weighing method,local capillaries were observed by micrangium perfusion.Results The intracerebral hematoma of the rats was reduced significantly( P
3.Effects of hirudo extract liquor on the expression of HSP70 and TGF?-1 in experimental intracerebral perihematoma tissues
Wenbin WU ; Changlin HU ; Yousong YANG
Journal of Clinical Neurology 1997;0(06):-
Objective To study the effects of hirudo extract liquor(HEL) on the expressions of HSP70 and TGF?-1 in intracerebral perihematoma tissues of rats.Methods We established the experimental ICH models in Wistar rats by stereotaxical injecting quantitative collagenase(0.7 U collagenaseⅦ) into their left caudate nuclei.The rats continued to be treated with HEL(treatment group) or normal saline(control group) through intravenous injection by vena caudalis and the scores of neurologic impairment in two groups were evaluated every day.The slices of these samples at 3rd,6th and 10th day were stained by immunohistochemistry and the positive cells of HSP70 and TGF?-1 were counted by image analysis system,respectively.Results The scores of neurologic impairment in two groups were remarkably reduced with time going((P
4.BIOMECHANICAL STUDY OF TRUNK MUSCLE STRENGTH AND INTERRELATED STUDY IN PATIENTS WITH LOW BACK PAIN CAUSED BY MILITARY TRAINING
Dehui ZHANG ; Changlin HUANG ; Jianqian WU
Medical Journal of Chinese People's Liberation Army 2001;0(11):-
To ascertain the change of trunk muscle strength and lumbar curvature and cross sectional area of M. sacrospinalis in low back pain caused by military training, the indexes of trunk muscle strength (PT/BW, TAE, F/E) were measured in patients with low back pain and healthy subjects with CYBEX 6000 isokinetic testing system. The lumbar curvature was measured in lumbar X ray films on the lateral projection in standing position, and the cross sectional area of sacrospinalis was measured by ultrasonography. All of the indexes were compared between the two groups.The results showed PT/BW of flexors was not significantly different between the patients and healthy subjects, TAE of flexors in patients was lower than that of healthy subjects ( P
5.The change in trunk muscle performance in recruits with low back pain caused by military training and its prevention
Dehui ZHANG ; Changlin HUANG ; Jianqiang WU
Medical Journal of Chinese People's Liberation Army 1981;0(04):-
To elucidate the change in trunk muscle performance in patients with low back pain caused by military training. The indices of trunk muscle strength (PT/BW, TAE, F/E, ER), the cross-sectional area of sacrospinalis, the amplitude and the duration of EMG, and the lumbar curvature were measured in recruits with low back pain after military training (n=40)and healthy subjects(n=40).All of the indices were compared between two groups. It was found that except PT/BW and ER of flexors and cross-sectional area of sacrospinalis, there were significant differences between other indices of the patients compared with healthy subjects(P
6.Research on experimental intracerebral hemorrhage with buflomedil hydrochloride injection
Wenbin WU ; Changlin HU ; Yongling ZHENG
Journal of Clinical Neurology 1988;0(02):-
Objective To observe the effect and safety of buflomedil hydrochloride injection to intra cerebral hemorrhage in rats. Methods We established the experimental intracerebral hemorrhage (ICH) model in rats by stereotactic injecting quantitative collagenase into their left caudate nucleuses.The effects of buflomedil hydrochloride injection on hematoma volume,the score of neurological function deficit signs, as well as histopathological changes were observed. Local capillaries were observed by micrangium perfusion.Results 5 days after buflomedil hydrochloride injection might alleviate Wister rats' neurological function deficit signs, 86% rats from Grade Ⅲ to Grade Ⅰ, 14% rats from Grade Ⅲ to Grade 0( P
7.The research of peripheral blood regulatory T cell in patients with idiopathic thrombocytopenic purpura
Yi ZHU ; Changlin WU ; Jiancheng XUE
International Journal of Laboratory Medicine 2008;29(6):490-491
Objective To investigate the change of peripheral blood CD4+CD25+ regulatory T cells (Tr) in patients with idiopathic thrombocytopenic purpura (ITP), and to analyze its role in the pathogenesis of ITP.Methods Anticoagulated venous blood was collected from ITP patients (ITP group,n=35) and healthy controls (healthy control group,n=35). T lymphocytes were isolated and purified with human CD3+ T cell enrichment columm. The percentage of peripheral blood CD4+CD25+ regulatory T cells was detected by immunofluorescence staining (PE-anti-CD4 monoclone antibody and FITC-CD25 monoclone antibody) and bicolor flow cytometry by CD25/CD4 gating.Results The number and constituent ratio of CD4+CD25+T cell were significantly lower in ITP group than those of healthy control group (P<0.05).Conclusion There is peripheral blood celluar immunological function disorder in ITP patients, and decrease of CD4+CD25+ T cell population may be involved in the pathogenesis of ITP.
8.Effect of tanshinoneⅡ A on expression of protein S100A1in acute myocardial ischemia rats
Maolin WU ; Changlin ZHAI ; Yamei ZHANG ; Feifei WU ; Yingzhi ZHANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2014;(5):350-353
ObjectiveTo investigate the protective effect of tanshinoneⅡA on the expression of S100A1 protein after acute myocardial ischemia injury in rats.Methods Sixty Wistar rats were randomly divided into sham operation group, acute myocardial ischemia model group and tanshinoneⅡA pretreatment group by random number table. The acute myocardial ischemia model was established by thoracotomy and penetration of a thread and occlusion around the root part of the left anterior descending coronary artery, while the sham operation group was established only by thoracotomy and penetration of a thread around the root part of that artery but without occlusion; 3 days before the operation, in the tanshinoneⅡA pretreatment group, intraperitoneal injection of tanshinoneⅡA solution(at a dose of 1.5 mg/kg) was applied, while in the sham and acute myocardial ischemia groups, intraperitoneal injection of an equal volume of saline was given. Myocardial cell apoptosis was detected by the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL), the levels of serum superoxide dismutase (SOD), malondialdehyde(MDA), creatine kinase(CK), lactate dehydrogenase(LDH) and S100A1 protein were examined and the level of expression of S100A1 protein in myocardial tissue was assayed by immunohistochemical staining and Western Blot.Results Compared with the sham operation group, the myocardial cell apoptosis rate, the contents of MDA, CK, LDH, S100A1 and the level of S100A1 expression in myocardial ischemia group and tanshinoneⅡA pretreated group were significantly increased, while SOD activity was decreased obviously; compared with the myocardial ischemia model group, the myocardial cell apoptosis rate, the contents of MDA, CK, LDH, S100A1 and the level of S100A1 protein expression were significantly reduced〔apoptosis rate:(32.1±4.2)% vs.(72.4±5.4)%, MDA(μmol/L): 9.1±2.2 vs. 17.3±5.2, CK(U/L): 83.3±12.2 vs. 107.5±12.4, LDH (μmol·s-1·L-1): 84.0±16.4 vs. 114.4±16.0, S100A1(μg/L): 37.6±6.0 vs. 78.4±8.6,P<0.05 orP<0.01〕, while the activity of SOD was increased markedly in tanshinoneⅡA pretreated group(kU/L:72.8±10.2 vs. 49.6±8.8,P<0.01). TUNEL staining showed that in the myocardial ischemia model group and tanshinoneⅡA pretreated group, the myocardial cells represented positive staining(brown-yellow in color), irregular in shape with nuclear pyknosis, cell detachment from the surrounding tissue and other characteristics. And in sham operation group,the staining of majority of cells was negative. The results of immunohistochemistry showed that S100A1 protein staining was relatively deep in the myocardial ischemia model group and tanshinoneⅡA pretreated group, and in the latter group, the color of S100A1 protein positive staining was not as deep as that in the former group. Western Blot showed that the S100A1 protein expression in myocardial ischemia model group was 2.8 folds of that of the sham operation group, while the S100A1 protein expression in tanshinoneⅡA pretreated group was significantly decreased compared with that of myocardial ischemia model group(bothP<0.05),which was 1.5 folds of that of the sham operation group.ConclusionTanshinoneⅡA may play a role in inhibiting the expression of S100A1 protein to protect against acute myocardial ischemia injury, suggesting that this agent have a potential effect for treatment of myocardial ischemia.
9.Relationship of serum cystatin C level with cytokines and carotid atherosclerosis in mintenance hemodialysis patients
Bibo WU ; Liming ZHANG ; Changlin MEI ; Qi TANG ; Yong YU
Chinese Journal of Nephrology 2011;27(11):802-806
Objective To investigate the serum cystatin C (CysC) level and explore its relationship with cytokines and atherosclerosis (AS) in maintenance hemodialysis (MHD) patients.Methods A total of 110 stable MHD patients undergoing hemodialysis for at least six months and 60 healthy control people were enrolled in the study.Serum levels of CysC and high-sensitivity Creactive protein (hsCRP) were measured by immunoturbidimetry.The serum levels of total homocysteine (tHcy),IL-1β,IL-6 and TNF-α were determined by ELISA.Prevalence of atherosclerosis was detected by carotid ultrasonography.The relationship of CysC level and cardiac geometry incidence in MHD patients was analyzed by Logistic regression model.Results The serum CysC level was significantly higher in MHD patients as compared with healthy controls [(6.19±0.95) mg/L vs (0.76±0.21) mg/L,P<0.01],and the serum levels of hsCRP,tHcy,IL-1β,IL-6,TNF-α were significantly higher in MHD patients than those in healthy control group (P<0.05 or P<0.01).The serum CysC level was higher in MHD patients with carotid artery atherosclerosis compared to patients without carotid artery atherosclerosis (P<0.05).CysC was positively correlated with hsCRP,tHcy,IL-1β,IL-6,TNF-α respectively (P<0.05 or P<O.01),and was positively correlated with carotid intimal medial thickness (IMT) and AS.Besides,a negative correlation was found between the serum CysC level and the serum albumin level (P<0.05),while CysC was positively correlated with dialysis duration,systolic pressure and iPTH (P <0.05).Conclusion Serum CysC level is significantly higher in MHD patients and is correlated with hsCRP,tHcy,IL-1β,IL-6,TNF-α as well as carotid artery atherosclerosis,which indicates that CysC is an independent risk factor of AS in MHD patients.
10.Heart protection of L-carnitine in chronic renal failure rats
Liming ZHANG ; Qi TANG ; Changlin MEI ; Yizhou LU ; Bibo WU
Chinese Journal of Nephrology 2012;28(5):377-382
Objective To investigate the effect of L-carnitine on pathological changes of myocardium and the underlying mechanism in chronic renal failure rats (CRF). Methods A total of 55 male SD rats were randomly divided into sham group (n=10),model group (n=15),low dose (300 mg/kg),medium dose (600 mg/kg) and high dose (900 mg/kg) L-carnitine group(n=10,each).5/6 subtotal nephrectomy was performed in these rats without sham group.One week after the operation,normal saline or corresponding dose L-carnitine were intragastrically administrated to sham and model group or L-carnitine groups for 17 weeks.Transthoracic echocardiography,mean arterial pressure (MAP),heart rate (HR) and heart weight/body weight were assessed.Moreover,24h urine protein,renal function,SOD,MDA,IL-6,ATP,ADP were measured at the end of the study.Additionally,pathological changes in myocardium were detected by light microscope and transmission electron microscope. Results (1) ATP (μmol/g·wt)in L-carnitine groups (2.35±0.24,3.59±0.28,3.78±0.25) was significantly higher than that in model group (1.61±0.12) (all P<0.01).(2) Thickness of posterior wall of left ventricle (mm) in high dose L-carnitine group was thinner than that in model group (3.74±0.23 vs 4.18±0.48,P<0.05). (3) The ratios of heart weight to body weight in both medium dose and high dose L-carnitine groups (3.92±0.27,3.65±0.2) were significantly lower compared to model group (3.99±0.27) (all P<0.01). (4) Under light microscopy,disarrangement and hypertrophy of cardiac myocytes,increased myocardial fibrosis were observed in model group, while these changes and the pathological scores were significantly improved in both medium dose and high dose L-carnitine groups (7.14±1.07,6.13±0.99),as compared with model group (9.88±1.13) (all P<0.01).Under electron microscopy,typical changes in cardiac hypertrophy were observed,including dissolution of myocardial fibers,increasing and swelling of mitochondria,membrane rupture as well as matrix increase in model group,while these changes were ameliorated by L-carnitine in a dose-dependent manner. (5) Seventeen weeks after the treatment,both IL-6 and MDA were decreased in all L-carnitine-treated groups than those in model group [IL-6 (ng/L):261.86±13.18,240.12±18.7,233.34±36.88 vs 596.64±81.41; MDA (nmol/L):15.23±2.01,12.41±0.6.10.97±1.9 vs 21.84±2.71).Whereas,SOD (U/ml) were increased in L-carnitine-treated groups (51.2±6.11,58.51±5.52,60.63±6.94) than that in model group(32.01 ±5.69 )(all P<0.05).(6) No significant differences of systolic,diastolic blood pressure or MAP were found among groups. Conclusion L-carnitine can improve energy metabolism,micro-inflammation and oxidative stress in myocardium of CRF rats,which may be associated with the amelioration of cardiac hypertrophy and fibrosis.