BACKGROUND: Nitric oxide synthase(NOS) is the key factor for the synthesis of nitric oxide (NO) . Because NO combines with oxygen, hemoglobin and other substances in vivo easily and deactivates quickly, and it is not exactly determined, so determining the activity of NOS is the important link for further studying the pathogenesis of NO in cerebral ischemia/reperfusion (I/R)injury.OBJECTIVE: To study the effect of different types of NOS in the process of cerebral I/R injury.DESIGN: Randomized and controlled animal experiment.SETTING: Instituteof Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: This experiment was carried out in the Shandong Key Laboratory of Prevention and Treatment for Encephalopathy from May to December 2005. Twenty-eight adult healthy male Wistar rats, of clean grade, weighing from 220 to 260 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomly divided into sham-operation group (n =4) and cerebral ischemia group (n =24). Six time points were set in cerebral ischemia group: ischemia 1 hour reperfusion 6 hours, 12 hours, 1 day, 3 days, 7 days and 14 days, 4 rats at each time point.METHODS: Rat models of middle cerebral artery occlusion/reperfusion were established by suture-occluded method through inserting a suture into the left internal-external carotid artery. The expressions of different types of NOS at different time points after cerebral I/R were detected by immunohistochemical technique.MAIN OUTCOME MEASURES: ①Toluidine blue-stained two groups of nerve cells; ② The expression and distribution of neuronal NOS (nNOS), endothelial NOS (eNOS) and inducible NOS(iNOS) at different time points.RESULTS: ①Karyopyknosis and cell debris appeared in the nerve cells of the injured region of cerebral ischemia group,and there were no significant differences of cells among different time points. ② Six hours after reperfusion, the expressions of nNOS, eNOS and iNOS were found in the neurons of brain tissue and increased with the elongation of time of reperfusion. The regions in which different types of NOS in neurons of brain tissue were expressed were cortical area and corpora striata. nNOS and iNOS were highly expressed within 12 hours to 7 days after reperfusion in the brain, and eNOS was highly expressed within a short time period, i.e. 6 hours to 3 days after reperfusion. eNOS expression increasing and decreasing occurred earlier than nNOS and iNOS. But the expressions of three kinds of NOS all reached peak on the first day after reperfusion. The changing tendencies of the expression of three kinds of NOS in the cortical area and corpora striata were the same basically.CONCLUSION: After cerebral I/R injury, the high expression of eNOS occurs early and lasts for a short time, while that of nNOS and iNOS occurs late and lasts for a long time.

Numb,one of the important cell fate determinants,plays a key role in regulation of nerve system development,and it is closely related to the occurrence of tumors.It is proven that Numb is implicated in the processes of occurrence,invasion and metastasis of tumors through regulating the way of cell division,cell polarity,epithelial-mesenchymal transitions and several signal pathways,e.g.,Notch,Hh,p53.The expression levels of Numb in cells are related to the malignant degree and prognosis of tumors.It is vital to explore the role of Numb in tumors for treating tumors at the standpoint of Numb.

Community rehabilitation of cardiovascular disease is of great significance. Exercise therapy of cardiovascular disease in communities is necessary and feasible. At present, the main problems China faced to carry out community exercise therapy of cardiovascular disease include: cardiovascular patients with a weak sense of community exercise therapy, lack of professional rehabilitation personnel, inadequate community-based exercise therapy practice and lack of community rehabilitation funding. To solve above problems, we should strengthen the community exercise therapy practice of cardiovascular disease, and promote the cooperation between community rehabilitation and hospital rehabilitation. In all, comprehensive efforts should be made in order to solve the problems.

BACKGROUND: It has been demonstrated that insulin-like growth factor-1 (IGF-1) is a kind of neurotrophic factor and protects from cerebral ischemia/reperfusion injury, the expression of IGF-1 is associated with the attack of ischemic stroke. The effects of IGF-1 and its receptor (IGF-1R) on neurobehavioral function are to be further studied.OBJECTIVE: To observe the effects of IGF-1 and IGF-1R on neurobehavioral function in rat models of cerebral ischemia/reperfusion injury.DESIGN: A randomized controlled observation.SETTING: Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University Medical College.MATERIALS: The experiments were carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases. Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University.METHODS: The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). The middle cerebral artery occlusion/reperfusion (MCAO/R) models were established by inserting a thread through left external-internal carotid arteries. The sham-operated rats were given the same treatments except inserting thread. ①Neurologic deficit test: The rats in the experimental group were assessed according to Bederson standard after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The sham-operated rats were assessed at corresponding time points; Without neurologic deficit was marked as 0 point; flexion of anterior claws as 1 point; unable to act against the pushing from the contralateral side as 2 points; circling while walking as 3 points; shaking as 4 points;unconscious mind as 5 points. ② Sample collection and treatment: The samples in the experimental group were collected after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion, and those in the sham-operated group ere collected at 24 hours postoperatively. The rats were anesthetized, brain samples were got at about 5 mm posterior to optic chiasma after brains were removed completely, then serial coronal sections (5 μm) were prepared, and 1 from 10 sections was stuck to the cover glasses treated with poly-L-lysine. ③ Morphological observation of neurons: The neurons in brain were observed by toluidine blue staining. ④ Detection of IGF-1 and IGF-1R: The expressions of IGF-1 and IGF-1R in cortex and striatum were detected with immunohistochemical technique, 4 fields were randomly selected to count the positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: ① The neurologic deficit; ② Morphological changes of neurons in brain; ③ Expressions of IGF-1 and IGF-1R in cortex and striatum.RESULTS: All the 28 rats were involved in the analysis of results. ① The neurologic deficit: The scores of neurologic deficit were (1.50±058) and (1.50±0.78) in rats after 7 and 14-day reperfusion, which were lower than that in rats after 6-hour reperfusion [(3.00±0.00), P ＜ 0.05]. ② Morphological changes of neurons in brain: The neurons in ischemic area appeared as paryopyknosis and became irregular in shape, there were obvious gaps around the cells, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ③ Expressions of IGF-1 and IGF-1R in cortex and striatum: The numbers of IGF-1 positive cells in cortex were (8.75±2.06), (11.13±1.14),(19.75±3.18), (17.38±3.11 ) and (11.23±2.28) respectively in rats after 6, 12-hours and 1, 3, 7-day reperfusion, which all were higher than that in sham-operated rats [(3.88±1.46), P ＜ 0.05], the numbers of IGF-1 positive cells in striatum were(8.25±2.21), (11.34±2.21), (18.23±2.64), (18.56±2.34) and (11.31±2.14) respectively in rats after 6, 12 hours and 1, 3, 7days reperfusion , which were also higher than that in sham-operated rats [(4.12±2.24), P ＜ 0.05]. The numbers of IGF-1R positive cells in cortex were (7.63±1.50), (10.50±2.34), (15.55±3.12), (15.37±3.01), (8.86±2.75) respectively in rats after 6, 12-hours and 1,3,7-day reperfusion, which all were higher than that in sham-operated rats [(4.13±1.81), P ＜0.05]. Those in striatum were (8.33±2.31), (10.24±2.09), (14.72±2.17), (14.24±2.77), (8.38±2.05), which were also higher than that in sham-operated rats [(3.76±2.35), P ＜ 0.05].CONCLUSION: The neurological function is damaged after cerebral ischemia/reperfusion, but it has a trend of self-recovery. The expressions of IGF-1 and IGF-1R are mainly distributed in cortex and striatum. Higher expressions of IGF-1 and IGF-1R maintain during 12 hours to 7 days after reperfusion and have a peak value at 1-3 days, which suggests that early expression of IGF-1 and IGF-1R are certain related to the recovery of neurological function.

BACKGROUND: Brain injury can induce the increased expression of basic fibroblast growth factor (bFGF) in brain,whereas FGFR is a very important player in the cell proliferation and differentiation, angiogenesis, skeletogeny, etc.OBJECTIVE: To observe the effect of bFGF and its receptor on neuronal apoptosis following cerebral ischemia/reperfusion injury in rats.DESIGN: A randomized grouping design and animal experiment.SETTING: Institute of Cerebrovascular Disease, Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University. Rabbit-anti-rat bFGF and fibroblast growth factor receptor-1(FGFR-1) monoclonal antibodies were provided by Wuhan Boster Biological Technology, Co.,Ltd.METHODS: The experiment was carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases.① The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). Models of middle cerebral artery occlusion/reperfusion (MCAO/R) were established by thread occlusion via left external-internal carotid arteries, and 4 rats in the experimental group were sampled at 1-hour ischemia/6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The rats in the sham-operated group were given the same treatment without inserting thread.After anesthesia, the brain was removed completely by cutting head, then the brain tissue at about 5 mm posterior to optic chiasma was cut down, then serial coronal sections (5 μm) were prepared. ② The brain tissues were stained with ematoxylin-eosin (HE), and the forms of neurons were observed under microscope. ③ TdT-mediated dUTP-biotin nick end labeling (TUNEL) method: there were buffy granules in nucleus which was positively stained (apoptosis). Four fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400). ④ The sections were stained with rabbit-anti-rat bFGF and FGFR-1 monoclonal antibodies, 4 fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: Apoptosis and the expressions of bFGF and FGFR-1.RESULTS: All the 28 rats were involved in the analysis of results. ① In the experimental group, the neurons in the ischemic sites were obviously decreased, some neurons appeared as paryopyknosis and became irregular, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ② In the sham-operated group, there were a few apoptotic neurons in the brain tissue, and the apoptotic neurons were obviously increased after ischemia, which mainly observed in cortexes and striatums of frontal and paritetal lobes. In the experimental group, apoptotic cells in cortexes began to increase gradually at 6 hours, and there were more cells at 12hours and 3 days, which reached the peak value at 1 day, and began to decrease at 3 day, but there were still more apoptotic cells at 14 days than in the sham-operated group. The number of apoptotic neurons and the changing trend in striatums were generally the same as those in cortexes (P ＞ 0.05). ③ In the sham-operated group, there were weak bFGF expression in the neurons of brain tissue, but there were fewer lightly stained positive cells. After cerebral ischemia, the bFGF expressions were increased, mainly observed in cortexes and striatums. The bFGF expression appeared at 6 hours after cerebral ischemia/reperfusion, and the number was increased gradually and deeply stained as the time of reperfusion prolonged (Figure 3), it reached the peak value at 1-3 days, and then weakened gradually, but it was still higher than in the sham-operated group at 14 days [(5.01 ±1.71), (5.21 ± 1.62) cells/visual field; (2.03± 1.73),(2.46± 1.38) cells/visual field, P ＜ 0.05]. ④ In the sham-operated group, lightly stained FGFR-1 positive cells could be observed in brain tissue. At 6 hours after cerebral ischemia/reperfusion, the FGFR positive cells began to increased in cortexes and striatums, which were the most at 1-3 days, and gradually decreased after 3 days, and the number was still a little more than that in the sham-operated group at 14 days [(5.01± 1.41), (5.20± 1.33) cells/visual field; (2.25±1.67),(2.32± 1.61 ) cells/visual field].CONCLUSION: After cerebral ischemia/reperfusion, the expressions of endogenous bFGF and FGFR-1 may be activated in cortex and striatum, then inhibit the neuronal apoptosis, and play its neuroprotective role.

OBJECTIVE:To explore the effect of nifedipine combined with losartan on the blood pressure and renal function of hypertension complicated with coronary heart disease. METHODS:150 patients with hypertension complicated with coronary heart disease were randomly divided into observation group and control group. Control group was orally treated with Nifedipine sustained release tablet 30 mg,once a day;observation group was additionally treated with Losartan potassium capsules 50 mg,once a day. The treatment course for both groups was 1 month. Systolic blood pressure,diastolic blood pressure,renal function indicators be-fore and after treatment,and incidence of adverse reactions in 2 groups were observed. RESULTS:After treatment,systolic blood pressure and diastolic blood pressure after 3 months was lower than 1 month and lower than before treatment in same group,renal function indicators were significantly lower than before treatment,and observation group was lower than control group,the differ-ences were statistically significant(P<0.05);the incidence of adverse reactions in observation group was significantly lower than control group,the difference was statistically significant(P<0.05). CONCLUSIONS:Nifedipine combined with losartan can well control the blood pressure of hypertension complicated with coronary heart disease,protect renal functions,with good safety.

Objective:To investigate the effect of short-term intensive treatment with insulin on metabolic control,?-cell function and insulin resistance in newly diagnosed type 2 diabetic patients. Methods:Thirty-eight newly diagnosed type 2 diabetic patients were randomly divided into two groups and treated with continuous subcutaneous insulin infusion(CSII) or multiple daily insulin injection(MDI) for 1 month.Fasting blood glucose(FBG) and lipid profiles were measured,and oral glucose tolerance test(OGTT)was performed before and after the treatment.The second phase insulin and C peptide secretions were measured by calculating area under curve(AUC) in OGTT.HOMA model was used to evaluate ?-cell function((HOMA-?)) and insulin resistance. Results:Excellent control of FBG was achieved after 1 month intensive treatment.(HOMA-?) was improved significantly. Conclusion:The excellent metabolic control and ?-cell function improvement can be achieved by short-term intensive insulin therapies like CSII or MDI.

Objective To investigate the neuroprotective effects and mechanism of phycocyanin in cerebral ischemia reperfusion injury in rats.Methods The model of middle cerebral artery occlusion reperfusion(MCAO/R) was established using the intraluminal filament occlusion with healthy adult male Wistar rats treated by phycocyanin.The apoptosis and the expression of bFGF and FGFR-1,IGF-1 and IGF-1R,iNOS and SOD were respectively determined by TUNEL and immunohistochemical staining to evaluate the effects of phycocyanin on above indexes.Results ① The rats showed neurobehavioral function disorders and the number of nerve cells reduced while apoptotic cells increased in ischemic area after ischemic reperfusion.In phycocyanin group,the number of apoptotic cells reduced siginificantly during reperfusion 12h~3d and the neurobehavioral function was better than that those in control group during reperfusion 7~14d.② In control group,the expressions of bFGF and FGFR-1 increased successfully from reperfusion 6h and reached a maximum at 1d,then subsided gradually in cortex and striatum.In phycocyanin group,the numbers of bFGF and FGFR-1 positive cells were higher than those in control group at the same time-points,which were significantly at reperfusion 1d and 12h~3d respectively.③ The expressions of IGF-1 and IGF-1R increased in cortex and striatum following cerebral ischemic and reperfusion.In phycocyanin group,the numbers of IGF-1 and IGF-1R positive cells in each time-point were higher than those in control group,which was significantly during reperfusion 6h~1d.④ In cotex and striatum,the iNOS and SOD expressed strongly and keep high level during 6h~7d with the maximum at reperfusion 1d.In phycocyanin group,iNOS expressed significantly higher during 12h~1d whlie SOD lower during 6h~1d than those in control group at the same time-point.Conclusion Phycocyanin might play a intrinsic antioxide effect by up-regulating SOD and down-regulating iNOS to inhibit neuronal apoptosis,and enhance the neuronal repairation by means of inducing the expressions of bFGF and IGF-1 following cerebral ischemic reperfusion in rats.

Objective To investigate the effects of lower limb training at different intensities on the recovery of walking function after stroke.Methods Thirty-six stroke patients were randomly divided into a 40-minutes of training group,an 80-minutes of training group and a 120-minutes of training group.Because of three missing followup cases,the final numbers of cases were 12,11 and 10 cases respectively.All of the patients received lower limb training based on regular rehabilitation therapy,but at the three different intensities:40 minutes,80 minutes or 120 minutes a day,five days per week,for 4 weeks.Walking function was assessed using the Holden functional ambulation categories (FACs) and the Fugl-Meyer assessment (FMA).These were carried out before treatment,and after two and four weeks of treatment.Results Before treatment,the average FACs of the three groups were (1.83 ±0.94),(1.73 ± 1.01) and (1.80 ± 1.03) respectively.Their average FMA scores were (19.17 ±5.52),(23.00±4.71) and (19.40 ±7.90).After two weeks of treatment,the average FAC in the 120-minutes training group was (3.30 ± 0.48),significantly higher than in the 40-minutes training group.After four weeks the average FACs in the 40-minutes,80-minutes and 120-minutes training groups were (2.67 ± 0.65),(3.18 ± 0.60) and (3.80 ±0.42) respectively.The differences between the 120-minutes group and the 80-minutes group as well as between the 80-minutes group and the 40-minutes training group were statistically significant.The average FMA scores of the three groups after four weeks were (25.08±4.46),(28.64±3.56) and (25.90±5.19) respectively.All the differences were significant compared with pre-treatment.There were no significant differences in FMA scores among the three groups after two weeks or four weeks of treatment.Before treatment,the proportion of patients able to walk independently in the 40-minutes,80-minutes and 120-minutes training groups were 16.7％,18.2％ and 20.0％.After two weeks the proportions had risen to 33.3％,36.4％ and 100％ respectively,so the results in the 120-minutes training group were significantly better.After four weeks of treatment the proportions of the three groups were 58.3％,90.9％ and 100％.All these improvements were significant compared with pre-treatment,but when compared with the 80-minutes training group,neither of the others showed a significant difference.Conclusions Intensive training can accelerate the recovery of walking function of patients after stroke and promote their ability to participate in daily activities.

Objective To investigate the change of transmembrane potential during Noradrenaline's early preconditioning phase and probe the mechanism of action.Methods Primary culture myocardial cell of neonate rat were divided into three groups at random.GroupⅠ is control group,Group Ⅱis treatment group by hypoxia,Group Ⅲ is treatment group by NA.Group Ⅰ didn't give any drug to culture for 40minutes;Group Ⅱ treated for 40minutes by hypoxia,Group Ⅲ exposed to Noradrenaline(0.2?g/ml)for 10 minutes then treated with hypoxia for 40 minutes.The fluorescence intensity of rhodamine-123 marked mitochondria were detected by flow cytometry.Results The fluorescence intensity of three Group were test by analysis of variance(F=461.3,P