Objective To study the transcription level of exogenetic glial cell line-derived neurotrophic factor(GDNF)in rat bone marrow mesenchymal stem cells(BMSCs).Methods The rat BMSCs were isolated and cultured in vitro,and randomly divided into three groups:GDNF-transfected group,blank plasmid-transfected group and non-transfected group.GDNF-transfected group and blank plasmid-transfected group were infected by the recombinant adenovirus of GDNF gene and blank plasmid,respectively.Each of the groups was divided again into 4 subgroups according to the cultured time(3d,6d,9d and 12d)after infection.The transcription level of GDNF gene in BMSCs was tested by quantitative real-time PCR.Results Blank plasmid-transfected group and non-transfected group had no expression of GDNF mRNA,and the relative level of GDNF mRNA transcription was 0.00751?0.00067 in GDNF-transfected group during the following 3~12 days after transfection.Conclusion Gene transfection technoqie mediated by adenoviral vector can transfect GDNF gene into BMSCs with the expression of exogenetic GDNF mRNA.BMSCs transfected with GDNF gene will be used for further research on therapy for the disease of central nervous system.

BACKGROUND: In the central nervous system(CNS) of normal adults there are neural stem cells or neural progenitor cells, which are capable of self-renewing and multiple differentiating. In normal physiological conditions, intrinsic neural stem cells are in a resting state. What state will they be in during cerebral ischemia?OBJECTIVE: To observe the distribution, proliferation and differentiation of intrinsic neural stem cells in focal transient ischemia in rats.DESIGN: A randomized controlled exploratory trial based on the rats.SETTING: Neurobiological department, histological and embryological department of a medical college.MATERIALS: The experiment was conducted in the Neurobiological Department of Luzhou Medical College from July 2001 to July 2002. Altogether 41 healthy adult SD rats of either gender, weighting 250 g - 300 g, were selected.INTERVENTIONS: The focal ischemia model was made by blocking middle cerebral artery(MCA) and reperfusing for 0.5 hour, 3 hours, 6 hours, 12 hours, 1 day, 2 days, 3 days, 5 days and 10 days. Sham-operation group was treated by the same method, but the filament was not long enough to block MCA, and normal rats served as control group. The rats were sacrificed at given time points, and their brains were made into cerebral slices. The single-and double-labeled immunohistochemical staining was employed to detect the proliferation, distribution and differentiation of intrinsic neural stem cells.MAIN OUTCOME MEASURES: The distribution, proliferation and differentiation of intrinsic neural stem cells.RESULTS: Immunoreactivity of proliferating cell nuclear antigen(PCNA)was present in most ependymal cells in ventricular zone(VZ), and PCNA-positive cells were sparsely distributed in the parenchyma in normal and sham-operation groups. At 3 hours of reperfusion, PCNA-labeled cells were first detected in rostral subventricular zone. At 12 hours of reperfusion and onward, PCNA-positive cells appeared in some choroid plexus cells in bilateral lateral VZ. At day 3 to day 10 of reperfusion, PCNA-labeled cells significantly increased in infarct boundary in preoptic area, striatum and deep layer of frontoparietal cortex. PCNA-labeled cells were first detected in subgranular zone of dentate gyrus 3 days after reperfusion, and increased with time. A very small number of double-positive cells expressed with PCNA and glial fibrillary acidic protein(GFAP) were first detected in infract boundary in preoptic area on day 3 and onward. No double-PCNA and NF-positive cells were detected within 10 days of reperfusion.CONCLUSION: Focal cerebral ischemia activates intrinsic neural stem cells, which proliferate and differentiate, and migrate toward ischemic striatum and frontoparietal cortex. This may help clarify the mechanism of functional recovery after ischemia.

BACKGROUND: There is few studies addressing the long-playing dynamic observation of cysteinyl aspartate specific protease 3 (Caspase-3) expression following cerebral ischemia/reperfusion.OBJECTIVE: To investigate the effect of transplantation of the neural stem cells (NSCs) modified with gene of glial cell line-derived neurotrophic factor (GDNF) on expression of Caspase-3 in adult Sprague Dawley rats with transient cerebral ischemia.DESING: Randomized controlled animal study.MATERIALS: Sixty Sprague Dawley rats were divided randomly into normal control group (N, n =5), ischemia/reperfusion group (IR, n=5), neural stem cell group (NSCs, n=25) and NSCs modified with gene of GDNF group (GDNF/NSCs, n =25). Several clean neonatal Sprague-Dawley rats were selected to harvest NSCs.METHODS: With the exception of normal control group, models of transient cerebral ischemia were created by modified suture method in other groups. At day 3 following reperfusion, 20 μL NSC suspension containing (4.0-5.0)×10~5 NSCs was infused into rats of the NSC group via right lateral ventricle. An equal volume of GDNF-modified NSC suspension was injected into rats of the GDNF/NSC group. 20 μL saline was infused into the rats of the ischemia/reperfusion group. Animals were anesthetized and sacrificed at week 1 following ischemia/reperfusion in the normal control and ischemia/reperfusion groups. Animals were anesthetized and sacrificed at weeks 1, 2, 3, 5, 7 following ischemia/reperfusion in the NSC and GDNF/NSC groups, 5 rats in each time point.MAIN OUTCOME MEASURES: The strept avidin-biotin immunostaining method was used to observe the distributive characteristics of Caspase-3 in the hippocampus and frontal parietal cortex.RESULTS: Immunohistochemical method (SP) showed that positive capase-3 products expressed in nucleus, cytoplasm and partial neurite. In hippocampus, number of Caspase-3-positive cells was decreased in NSC and GDNF/NSC groups. With the exception of at 1-week reperfusion, number of Caspase-3-positive cells was significantly lessened in the GDNF/NSC group compared with the NSC group at other time points (P < 0.05). In frontoparietal cortex, number of Caspase-3-positive cells was reduced in the NSC and GDNF/NSC groups over time. Except 1 and 2 weeks following ischemia/reperfusion, number of Caspase-3-positive cells was significantly lessened in the GDNF/NSC group compared with the NSC group (P < 0.05).CONCLUSION: Transplanting NSCs modified with gene of GDNF can improve remarkably neural function by deceasing Caspase-3 expression and reducing the nervous cell apoptosis. The transplantation of NSCs modified with gene of GDNF obtained better outcomes compared with NSC transplantation.

Objective To observe the effects of Sanjia Yigan Granule (S-JG) on hepatic fibrosis and explore its mechanism related to anti-lipid peroxidation in rats. Methods Forty male SD rats were randomly divided into four groups: normal group, model group, and compound Salviae Miltiorrhizae (Sm) group and S-JG group. Dimethylnitrosamine was injected intraperitoneally for 4 weeks to induce hepatic fobrosis. At the same time of modeling, Sm and S-JG were given in the corresponding groups. The rats were killed after four weeks. The histomorphylogic structure of the liver tissues was observed under optical microscope; the levels of MDA and SOD in serum were determined by radioimmunoassay. Results Compared with those in normal group, the collagen area in Masson staining and level of serum MDA were inreased obviously, and the level of serum SOD was dereased obviously in model group (P

Objective To investigate the brain glucose metabolism with 18 F?FDG microPET/CT in mouse models of intracerebral hemorrhage (ICH). Methods A total of 12 healthy adult male mice were randomly divided into sham operation group (group A, n=6) and ICH model group (group B, n=6) by simple random sampling method. The animal models were established by injecting collagenase Ⅳ into the caudate nucleus of mice. Thereafter (5.5±0.3) MBq of 18F?FDG was injected into caudal vein at 6 h, 24 h, 48 h and 3 d, 5 d, 8 d, 14 d, respectively, following anesthesia. 18 F?FDG microPET/CT scans were ac?quired 30 min after the trace injection. SUV in the perihematomal brain tissue of ICH was measured and an?alyzed. Two?sample t test was used to compare SUV between groups. Results ( 1) Some mice had mild neurologic deficit after the sham operation in group A, while all mice had a marked neurologic deficit in group B, especially at 24 h after 18 F?FDG injection. ( 2) After 6 h, FDG uptake in perihematomal brain tis?sue decreased(SUV=0.80±0.04), which significantly lower than that in the opposite side(SUV=1.10± 0?04;t=2.69, P<0.05) and decreased to the minimum at 24 h(SUV=0.50±0.05). 18F?FDG uptake in perihematomal brain tissue began to increase at 3 d(SUV=1.20±0.05) and kept increasing during the 14 d observation. Compared with the group A, glucose metabolism in group B was significantly lower at each time point(t=37.67-86.60, all P<0.05). Conclusions 18 F?FDG microPET/CT may dynamically reflect the changes of brain glucose metabolism in ICH mouse models. The FDG uptake in the center of ICH may disap?pear and the volume of hematoma with decreased uptake may shrink during the observation period.

BACKGROUND:Previous studies have demonstrated that cell transplantation has neuroprotective effect on intracerebral hemorrhage,and some researches have indicated that transplantation of bone marrow stromal cells(BMSCs)can promote neural function recovery after cerebral infarction.OBJECTIVE:To explore whether transplantation of BMSCs-modified by gtial cell line-dedved neurotrophic factor gene(GDNF)gene provides a better therapeutic effect than native BMSCs after stroke.METHODS:Totally 36 SD rats were induced intracerabral hemorrhage models by injecting autologous arterial blood,and then divided into 3 groups(n=6).each group was assigned into 2 sub-groups Rabbits in each group were stereotaxically grafted with 20 μL GDNF/BMSCs,BMSCs or saline respectively.The rats were executed at 1 and 2 weeks after operation,and immunohistochemistry was used to observe the expressions of synaptophysin(Syn)and growth associated protein-43(GAP-43)in the margin of the hemorrhagic focus.RESULTS AND CONCLUSlON:Compared with the BMSCs and control groups.both Syn-immunoreactive and GAP-43-immunoreactive products were significantly increased in the GDNF/BMSCs group(P＜0.05).Present results demonstrate that transplantation of GDNF gene-modified BMSCs provides better neuroprotection than native BMSCs delivery for stroke.

Objective To study the effect of DNA damage induced by H2 O2 on the micronucleus frequency in lymphocytes. Methods Resting lymphocytes were treated with different levels of H2 O2 (10,50,100,1 000 μmol/L).1 000 μmol/L H2 O2 was added into mitogen-stimulated lymphocyte cultures at different time intervals.Then micronucleus rate was examined by the conven-tional culture method.Results There was no significant change of the micronucleus frequency in the experimental groups.Conclu-sion H2 O2 could induce lymphocyte DNA damage rapidly,but exerts no effect on the formation of micronuclei,which may be relat-ed to the type of DNA damage and rapid DNA repair.

Previous experiments has shown that Ginsenoside Rb1 (GRb1), which is one of the most important active ingredients in ginseng (Panax ginseng C.A. Meyer), reduced infarct and neurologic deficit followed by the transient cerebral ischemia in rats. The mechanism of this neuroprotective function is unclear. In this study, we tested whether the neuroprotective effect of GRb1 is achieved through preventing the neuronal apoptosis and modulating expression of neuronal apoptosis inhibitory protein (NAIP). Focal cerebral ischemia was induced by the middle cerebral artery occlusion (MCAO) in Wistar rats. GRb1 (40 mg/kg, i.p.) was administered immediately after the onset of reperfusion. The rats with neurological deficits were randomly divided into 2 groups: the ischemia and the GRb1 group. Each group was again divided into subgroups according to the various reperfusion time (3 h, 12 h, 1, 2, 3, 5, 10 days, n=4 per time point). Apoptotic cells were analyzed using TUNEL. Immunohistochemical method was used to assess expression of NAIP. This results showed that the number of apoptotic cells elevated at 3 h of reperfusion, and peaked at 24 h, then declined, but the number of apoptotic cells at 10 d after ischemia was significantly more than those of control groups (P＜0.01). Compared with ischemia group, the apoptotic cells decreased at all subgroups of GRb1; however, the significant differences were only found from 12 h to 3 d of reperfusion. In normal and sham groups, NAIP weak immunostaining was diffusely present in the neurons of parenchyma. The number of NAIP-positive cells started to increase in ischemic regions at 3 h after ischemia, peaked at 12 h and declined up to 5 d of reperfusion. At 5 d after ischemia, the number of NAIP-positive cells was less than that of control group (P＜0.05). A few astrocytes strongly expressed NAIP in the ischemic area. In the GRb1 group, the number of NAIP-positive cells from 12 h to 10 d after ischemia was evidently higher than in the ischemia group. Thus, these results suggest that GRb1 has potential ability to prevent apoptosis, the mechanism of which is related to induce expression of NAIP.

OBJECTIVE:To investigate the effects of urapidil on the hemodynamics of the right heart in patients with rheumatic heart disease and secondary pulmonary hypertension METHODS:34 patients with rheumatic heart disease and varying degrees of secondary pulmonary hypertension were included in the study In all patients,the hemodynamic parameters and pressure of systemic circulation were determined before and 20,40,60 min after urapidil intraverous injection in a dose of 0 4mg/kg RESULTS:Pulmonary artery pressure,pulmonary vascular resistance,pulmonary capillary wedge pressure and systemic artery pressure were decreased after urapidil injection The reduction of resistance in the pulmonary vascular bed was greater than that in the systemic circulation There were no significant alterations in heart rate and cardic output No serious side-effects were observed in the study CONCLUSION:Urapidil may be a promising drug in the treatment of patients with pulmonary hypertension due to rheumatic heart disease

OBJECTIVE:To evaluate the analgesic efficacy and safety of tramadol im.on pain after thoracic surgery.MET_ HODS:34cases after thoracic surgery were given tramadol im.in a dose of1.5mg/kg.The analgesic effect and adverse effect were observed.RESULTS:The significant effective rate of pain relief was41.2%and effective rate was38.2%with a total effective rate of79.4%.Some adverse effects including temporary nausea,vomiting,perspiration,dysuria were observed in a part of the patients.No respiratory inhibition was found.CONCLUSION:Tramadol(1.5mg/kg)im.is safe and effective in treatment of the pain after thoracic surgery.