Matrix metalloproteinases (MMP) are a family of proteolytic enzymes that degrade various components of the extracellular matrix (ECM).Both animal expodments and clinical sample analysis have shown that balance in expression and activation of MMP and inhibition by tissue inhibitors of matrix metallo-proteinase (TIMP)is critical for the development of stenotic change.This review article focuses on the role of MMP relevant to restenosis after vascular injury.

The aim of our study was to observe the effect of diabetes melitus and hy- pertension on cerebral hemodynamics.The flow velocity of cerebral basal arteries were inves- tigated with transcranial Dopplersonography(TCD) in the group of diabetesmellitus,hyper- tension and diabetes combined with hypertension.The results that the flow abnormality in diabetic group(n=48) was41 .7% and in hypertensive group(n=1 6 0 ) was30 % ,respective- ly,butin the group(n=35 ) of diabetes combined with hypertension,the abnormality reae- hed as high as 82 .9% and the abnormal rate of the latest group was significantly higher than former two groups(P

Objective To study the pathway of IFN-?-iNOS-NO on T lymphocytic cells in the rats with experimental autoimmune myasthenia gravis (EAMG).Methods Lewis rats were divided randomly into groups EAMG , normal control(NC) and completed Freund's adjuvant control(CFA-C) . The rats were immunized in foot pad, abdomen and back subcutaneouly with the AChR protein extracted from electric organ of Narcine timilei and CFA in the EAMG group, or only with CFA in the CFA-C group. The above-mentioned emulsions were injected again after 4 weeks . At the 7th week after first immunization, T lymphocytic cells in every rat were separated from the spleen. After cultured in vitro for 48 h, the content of IFN-? in the supernatant of cultured T lymphocytic cells was detected by enzyme-linked immunoadsordent assay (ELISA), and the level of NO was examined with Griess reagent method. Results The contents of IFN-? were (81.68?10.23 ) pg/ml in EAMG group,(29.20?5.41) pg/ml in NC group,(31.54?6.12) pg/ml in CFA-Cgroup.The levels of NO were (23.68?7.13 )?mol/L in EAMG group ,(9.05?2.11) ?mol/L in NC group,(10.21?2.67 )?mol/L in CFA-C group.The contents of IFN-? and NO in EAMG group were increased markedly than groups NC and CFA-C(allP

Objective To investigate the role of the extracellular signal-regulated kinases 1 and 2 (ERK1/2) in rotenone-mediated dopaminergic cell damage. Methods Neuronal rat adrenal pheochromecytoma(PC12) cells treated with rotenone were used as the cell model of Parkinson' s disease (PD). Cell viability of PC12 cells after exposure to rotenone was detected by MTT (methyl thiazolyl tetrazolium) method. Immanohistechemistry was used to detect the phosphorylation of ERK1/2 in cells exposed to rotenone. Western blotting was used to verify the phosphorylation of ERK1/2 and to observe the effect of PD98059, an inhibitor of the upstream mitogen activated protein kinase kinase (MEK) that phosphorylates and activates ERK1/2, on rotenone-induced ERK1/2 phosphorylation and cell viability.Results The viability (represented by A570 of PC12 cells exposed to 5 μmoL/L rotenone) declined with the increase of exposure time from 1 h to 24 h (%, 1 h :75.46±5.47, 2 h : 70.42±1.94, 4 h : 65.23± 0.96, 8 h : 59.04 ± 2.85, 24 h :29.64 ± 1.63, comparison between different time points(F=143.014, P=0.000) ; compared with control groups(100.00±2.89), q value: 17.07, 20.58, 24. 19, 28.50, 48.95 respectively, all P <0.01). After exposure to rotenone, phosphorylated ERK1/2 aggregated in the PC12 cells. Western blotting indicated that rotenone induced a biphasic phosphorylation of ERK1/2, which increased from 30 min after rotenone treatment, reached the peak at 1-2 h, decreased at 4 h, and increased again at 8 h, and disappeared after 16 h; PD98059 significantly inhibited ERK1/2 phosphorylation induced by rotenone, and attenuated cell injury examined at 1, 2 and 8 h. Conclusions Our study suggested that ERK1/2 activation plays a detrimental role in rotenone toxicity, and raised the possibility that abnormal patterns of ERK1/2 activation may contribute to dopaminergic neuronal cell death in PD.

BACKGROUND: It is indicated in the study of recent years that gingko biloba extract (EGB) is a kind of natural cleaner of free radical and it protects the body from the damage induced by free radical and improves cerebral circulatory disturbance and neuronal function. But the experimental or clinical study on the effects of EGB on high neural functional activity, like cognition, is relatively lagged.OBJECTIVE: To probe into the function of EGB on high functional activity in central neural system so as to provide the experimental evidence on clinical application of EGB in treatment of cognitive disturbance.DESIGN: Randomized controlled experiment was designed.SETTING: Department of Geriatrics of Psychiatric Hospital affiliated to Tongji Medical College of Huazhong University of Science and Technology,Department of Pathophysiology in Tongji Medical College of Huazhong University of Science and Technology and Department of Neurology in Union Hospital affiliated to Jinan Medical College of Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Basic Department of Tongji Medical College of Huazhong University of Science and Technology in June 2002. Forty Wistar rats were employed and randomized into 5groups, named as normal control of aged rats (normal group), model group,EGB 75 mg/kg group, EGB 150 mg/kg group and EGB 500 mg/kg group, 8 rats in each one.METHODS: Scopolamine was used to induce disturbance of learning and memory in aged rats to simulate the model of senile dementia animals. In normal and model groups, physiological saline of same volume was used for gastric perfusion and in every EGB group, EGB of 75, 150 and 500 mg/kg was used for gastric perfusion successively, 50-400 g/time, continuously for 5 days. On the 6th day, water maze and evading-dark-room tests were performed. During the testing, the medical perfusion stopped. The assay methods of behavioral training of learning and memory, such as experiment with water maze and evading-dark-room test, and biochemical assay were used to observe the changes in learning and memory and in acetylcholine (Ach) and protein contents in cerebral hippocampus before and after medication.MAIN OUTCOME MEASURES: ① Time required in maze test of rats in each group. ② Mistakes in maze test of rats in each group. ③ Time required and mistakes in evading-dark-room test of rats in each group. ④Contents of Ach and protein in cerebral hippocampus of rats in each group.RFSULTS: Except that 1 rat was died without definite reason in EGB 150 mg/kg group and 1 rat was escaped in either EGB 75 mg/kg or 500 mg/kg group during gastric perfusion, terminally, 37 rats entered result analysis.① The time required and mistakes in maze test in every EGB group were less remarkably than model group (P＜0.05 or P＜0.01). The time required and mistakes in maze test in model group were higher remarkably than normal group (P＜0.01). ② In learning of passive escaping in evading-darkroom test, the duration of learning for the rats in EGB 500, 150, 75 mg/kg groups was shorter remarkably than that in model group [(156.78±25.97),(172.66±13.56), (198.54±17.12), (208.34±28.56) s, P ＜ 0.05 or P＜0.01].The mistakes of electric shock in EGB 500, 150, 75 mg/kg groups were less remarkably than model group [(3.41±0.26), (6.97±0.35), (7.23±0.62),(8.38±0.92) times, P＜0.01]. The times of electric shock in EGB 500 mg/kg group was less significantly than 150 mg/kg group (P＜0.01) and that in 150 mg/kg was less remarkably than 75 mg/kg group (P＜0.05). ③ Hippocampal Ach content in modeled rats in EGB 500, 150, 75 mg/kg groups was higher than that in model group [(421.89±36.32), (387.45±32.76),(380.17±41.25), (365.28±11.42) μg/g, P＜0.05 or P＜0.01]. Hippocampal Ach content in 500 mg/kg group was higher significantly than 150 and 75 mg/kg groups (P＜0.01). In addition, compared with normal group,protein content in hippocampus in rats with disturbance of learning and memory induced by scopolamine in model group was reduced significantly [(41.75±3.82), (95.13±6.34) mg/kg, P ＜ 0.01]. After administrated with EGB,even though the protein content in hippocampus was increased in experimental rats after modeling, the difference was not significant (P＞0.05).CONCLUSION: EGB improves significantly learning and memory in experimental animal in dose-dependence and increases significantly Ach content in hippocampus.

Objective To evaluate the efficacy and safety of pramipexole in treating restless legs syndrome (RLS).Methods A search for randomized,double-blind,and placebo-controlled clinical trials of pramipexole in treating moderate to severe RLS using CNKI,PubMed,Embase and Cochrane Library database was carried out. A meta-analysis of included clinical trials was performed with RevMan 5.0 software.The 2 outcomes that the weighted mean difference(WMD) of change from baseline in International RLS Study Group rating scale(IRLS) score and the relative risk (RR) of response based on the Clinical Global Impression-Improvement (CGI-I) scale score were calculated for efficacy.Safety was assessed with RR of the adverse event (AE).Results A total of 5 clinical trials were included in this meta-analysis,of which 1776 patients were randomly assigned (945 on pramipexole,831 on placebo).The records of patients were pooled.Overall WMD were - 6.34 ( Z =12.76,P ＜ 0.01 ) for the change from baseline in IRLS score,and RR of response based on CGI-I were 1.65 (Z =10.39,P ＜0.01).The overall RR of pramipexole versus placebo were 1.14 ( Z =1.87,P =0.06 ) for AE.Conclusion To treat RLS,pramipexole is an effective and safe drug.

Apoptosis of dopaminergic neurons in the nigrostriatal projection plays a crucial role in the pathogenesis of Parkinson's disease (PD). Although the detailed mechanisms responsible for dopaminergic neuron loss are still under investigation, oxidative stress is identified as a major contributor for neuronal apoptosis. In the current study, we studied the effects of MPP(+), a substrate that mimics oxidative stress, on neuron-like PC12 cells and the underlying mechanisms. PC12 cells were cultured and treated by 100 μmol/L MPP(+) for 4, 8, 16, 24 and 48 h, respectively. For drug pretreatment, the PC12 cells were incubated with N-acetyl-l-cysteine (NAC, 5 mmol/L), an antioxidant, SP600125 (20 μmol/L) or PD98059 (100 μmol/L), two pharmacological inhibitors of JNK and ERK1/2, for 1 h before addition of MPP(+). Cell apoptosis was measured by flow cytometry. The mRNA expression of Cu(2+)/Zn(2+)-SOD, GSH-Px, Bcl-2 and Bax was detected by RT-PCR. The protein expression of p-ERK1/2 and p-JNK was determined by Western blotting. Our results showed that MPP(+) exposure could induce substantial PC12 cell apoptosis. The pretreatment of SP600125 or PD98059 could effectively reduce the apoptosis rate by reducing the ratio of Bax/Bcl-2 mRNA levels. MPP(+) exposure also induced high level of reactive oxygen species (ROS), marked by dramatic increase of Cu(2+)/Zn(2+)-SOD and GSH-Px mRNA levels. The elevated ROS was strongly associated with the activation of JNK and ERK1/2 signal pathways after MPP(+) exposure, since the pretreatment of NAC significantly reduced the upregulation of p-JNK and p-ERK1/2. Finally, the pretreatment of SP600125, but not PD98059, alleviated the increase of Cu(2+)/Zn(2+)-SOD and GSH-Px mRNAs induced by MPP(+), suggesting that the activation of the JNK signal pathway, but not the ERK1/2 signal pathway, could, in some degree, antagonize the generation of ROS induced by oxidative stress. In conclusion, our results suggest that JNK and ERK1/2 signal pathways, which are activated via ROS, play a crucial role in neuronal apoptosis induced by oxidative stress.

The regulation of astroglia on synaptic plasticity in the CA1 region of rat hippocampus was examined. Rats were divided into three groups: the newly born (< 24 h), the juvenile (28-30 days) and the adult groups (90 - 100 days), with each group having 20 animals. The CA1 region of rat hippocampus was immunohistochemically and electron-microscopically examined, respectively, for the growth of astroglia and the ultrastructure of synapses. The high performance liquid chromatography was employed to determine the cholesterol content of rat hippocampus. In the newly-born rats, a large number of neurons were noted in the hippocampal CA1 region of the newly-born rats, and few astroglia and no synaptic structure were observed. In the juvenile group, a few astroglias and some immature synapses were found, which were less than those in adult rats (P < 0.01). The cholesterol content was 2.92 +/- 0.03 mg/g, 11.20 +/- 3.41 mg/g and 12.91 +/- 1.25 mg/g for newly born, the juvenile and the adult groups, respectively, with the differences among them being statistically significant (P < 0.01). Our study suggests that the astrocytes may play an important role in the synaptic formation and functional maturity of hippocampal neurons, which may be related to the secretion of cholesterol from astrocytes.
Age Factors ; Animals, Newborn ; Astrocytes/cytology ; Astrocytes/metabolism ; Astrocytes/*physiology ; CA1 Region, Hippocampal/*physiology ; CA1 Region, Hippocampal/*ultrastructure ; Cell Communication/physiology ; Cholesterol/metabolism ; Neuronal Plasticity/*physiology ; Random Allocation ; Rats, Wistar ; Synapses/*physiology ; Synapses/ultrastructure

Objective To study the effects of dendritic cells (DC) modified with transforming growth factor ?1 (TGF-?1) gene on experimental autoimmune myasthenia gravis (EAMG) in Lewis rats. Methods 30 female Lewis rats were divided randomly into 6 groups: normal group, EAMG group, DCs treatment group, pcDNA_3-TGF-?1-DCs treatment group, pcDNA_3-DCs treatment group and normal saline group. The rats were immunized with the acetylcholine receptor (AChR) protein extracted from electric organ of Narcine timilei and completed Freund’s adjuvant (CFA) in the experiment groups except normal group. 2?106 pcDNA_3-TGF-?1-DCs/rat were injected subcutaneously into the backs of the rats that had been immunized 5 days earlier with AChR+CFA. The rats in DCs treatment group, pcDNA_3-DCs treatment group and normal saline group were injected in parallel with untreated DCs, pcDNA_3-DCs and normal saline respectively. Then the clinical manifestations were observed everyday. And 7 weeks after the first immunization, repetitive nerve stimulation, detection of acetylcholine receptor antibody (AChRab) and ultrastructural study of neuromuscular junction (NMJ) were performed. Results (1) The mild symptoms were observed on 1 or 2 rats in the experiment groups except normal group after a week, which lasted for 2 to 5 days. After about 5 weeks, the rats in EAMG group, DCs treatment group, pcDNA_3-DCs treatment group and normal saline group presented some symptoms at different degree like myasthenia gravis, and only one of the rats in pcDNA_3-TGF-?1-DCs treatment group presented mildly decreased activity. (2) The significant decrement of repetitive nerve stimulation were found in EAMG group, DCs treatment group, pcDNA_3-DCs treatment group and normal saline group(16.75?6.13, 17.75?7.81, 18.25?8.22 and 16.50?7.14, respectively), but there was no attenuation in pcDNA_3-TGF-?1-DCs treatment group and normal group(3.20?3.70 and 5.60?2.70, respectively). The percentage of decrement in pcDNA_3-TGF-?1-DCs treatment group was lower than that in EAMG group(5.60?2.70 and 16.75?6.13, respectively,P0.05). (4) The combined AChRs in NMJ of the rats in pcDNA_3-TGF-?1-DCs group were higher than that in EAMG group, and the structure changes of the synapse were relieved.Conclusion It suggests that DCs, transfected with pcDNA_3-TGF-?1, when injected subcutaneously into Lewis rats with incipient EAMG, could inhibit the production of AChR-Ab, relieve the pathologic changes in NMJ and ameliorate the development of EAMG.

Objective To explore the effects of 14-3-3 γ gene on dopaminergic neurons of PD rat model.Methods 6 hydroxydopamine (6-OHDA) was injected into the corpus striatum of 60 SD rats to establish PD model.A week later,Ad/14-3-3 γ was injected into the corpus striatum of 16 rats,while PBS and Ad-LacZ were injected into the corpus striatum of 16 and 28 rats,respectively,as control.X-gal dyeing was utilized to examine the LacZ reporter gene expression in the corpus striatum at 3 day,2 week and 6 week.Real-time PCR was utilized to test the expression level of 14-3-3 γmRNA at 2 weeks after Ad/14-3-3 γ injection.Immunohistochemistry technique was used to detect TH positive neurons and fibers in the corpus striatum and substantial nigra.Western blotting was performed to check the expression of 14-3-3 γ in the corpus striatum at 2 weeks and caspase-3 at 6 veeks after Ad/14-3-3 γ injection.High performance liquid chromatography (HPLC) method was used to examine the contents of DA and DOPAC in the corpus striatum.The rats underwent rotational ethological examination at 1,2 and 6 weeks after the second injection.Results The expression of β-gal,which showed the successful LacZ gene transfection,was found in the corpus striatum of LacZ groups.The 14-3-3 γ mRNA and protein expression in the corpus striatum were significantly higher in the Ad/14-3-3 γ group than in the other two groups.The TH-positive cell ratio of substania nigra to contralateral area in the lesion side was 0.44±0.17 and the optical density (OD) of TH-positive fibers was 0.62±0.14 in the Ad/14-3-3 γ group,both higher than those in PBS group (0.16±0.13 and 0.36±0.15) and LacZ group (0.15±0.09 and 0.30±0.11) (all P＜0.01).The contents of DA and DOPAC in the lesion sides of corpus striatum were increased in the Ad/14-3-3 γ group [(248± 116)ng/g and (752±177) ng/g] than in PBS group [(106±35) ng/g and (724±159) ng/g] and LacZ group [(136±49) ng/g and (765±163) ng/g] (P＜0.01).The DA and DOPAC ratios of the lesion side of corpus striatum to the contralateral side were 0.37±0.14 and 0.38±0.17 in the Ad/14-3-3 γgroup,higher than those in PBS group (0.15±0.08 and 0.13±0.10,respectively) and LacZ group (0.19±0.11 and 0.16±0.09 (all P＜0.01).The expression level of caspase-3 was decreased in Ad/14-3-3 γ group than in PBS and LacZ groups at 6 weeks after the second injection.The turns/min induced by apomorphine in Ad/14-3-3 γ group were 9.4 ± 2.5 at 1 week after the Ad/14-3-3 γinjection,and reduced to 4.6±2.2 at 6 weeks later.While in PBS and LacZ group,the turns were 14.5±4.9 and 13.8±3.5 at 1 week after the second injection,6 weeks later rised to 18.7±5.2 and 20.6± 6.7 respectively,significantly higher than those in Ad/14-3-3 γ group (all P＜0.01).Conclusions 14-3-3γ gene transfer has a protective effect on the dopaminergic neurons and it may be a promising candidate gene for curing PD.