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Objective To observe the effect of aprotinin preconditioning on nitric oxide (NO), nitric oxide synthase (NOS) and oxyradical during spinal cord ischemia-reperfusion injury in rabbits.Methods 21 rabbits were randomly divided into the aprotinin treatment group (8 rabbits), control group (8 rabbits) and sham operative group (5 rabbits). The infrarenal segment in abdominal aorta was clamped for 60 min to construct the model of lumbosacral spinal cord ischemia in rabbits. Reperfusion was followed and kept on for 24 h until the blood flow regained normal. In the treatment group, aprotinin was given at 3×107 IU/kg as a short time intravenous injection for 10 min before ischemia, and then was drilled with micro pump by 1×107 IU/kg/h. Normal saline was used in the control group, the ischemia-reperfusion duration between aprotinin treatment group and control group remained same. The sham operative group was only exposured abdominal aorta and not clamped. The rabbits were killed before ischemia and 8 h, 24 h after ischemia-reperfusion, lumbar segment was harvested to detect content of NO, malondialdehyde (MDA), induced nitric oxide synthase (iNOS) and superoxide dismutase (SOD) of spinal cord.Results 8 h after spinal cord ischemia-reperfusion, compared with the control group, the content of NO, MDA and the activity of iNOS were less, and the activity of SOD was more in the aprotinin treatment group ( P<0.05).Conclusion Aprotinin pretreatment can reduce the content of NO, MDA and descend the activity of NOS. Moreover aprotinin pretreatment can ascend the activity of SOD and improve apoptosis of nerve cell.

【Objective】 To detect the copy numbers of brain-deriv ed neurotrophic factor (BDNF) gene in BDNF transfected PcDNA3.1(+)/BDNF/CHO cel ls with fluorescent quantitative polymerase chain reaction (FQ-PCR). 【Methods 】 BDNF DNA were amplified by GeneAmp 5700 Sequence Detection System with eq ual quantitative genomic DNA of PcDNA3.1(+)/BDNF/CHO, PcDNA3.1(+)/CHO and CHO cells as tamplates respectively. The process was repeated 30 times for every sam ples. The results were analyzed using q test. 【Results】 The copy numbers o f BDNF of PcDNA3.1(+)/BDNF/CHO cells and PcDNA 3.1(+)/CHO and CHO cells were 9 5 164±12, 31 622±10, 31 622±11 respectively. The copy numb ers of BDNF of PcDNA3.1(+)/BDNF/CHO cells were as three times as those of the P cDNA3.1(+)/CHO and CHO cells. The copy numbers of the two latters were the same . 【Conclusion】 The results clearly show that the PcDNA3.1(+)/BDNF/CHO cells h arbor two BDNF DNA copies.

Objective: To observe the effect of cilostazol on the ion channel of right ventricular cells in experimental rats, and to explore the ion channel mechanism of ciolstazol for preventing the ventricular arrhythmia in Brugada syndrome.
Methods: Our research was composed of 2 groups: ①Perfusion group, the cells were treated in 4 sub-groups by cilostazole at 1, 2, 5, 50μmol/L respectively, and there were 9, 5, 3, 7 cells were recorded at each sub-group to observe the differences of current density Ito at before and after treatment. ②Oral group, which included 4 sub-groups:Control 1 with 7 rats, Experiment 1 with 5 rats, and Control 2 with 8 rats, Experiment 2 with 6 rats respectively. The differences of current density Ito and ICa,L were studied between each Control and Experiment sub-groups.
Results: In Perfusion group,①with cilostazole 1, 2, 5, 50μmol/L treatment, the current density Ito decreased in all sub-groups, when the self-command voltage at+60mV, the Ito was signiifcantly different in each sub-group at before and after treatment, all P<0.05.②When each command voltage decreasing, the reduction rates of Ito were similar among 4 sub-groups, all P>0.05. In Oral group,①When the self-command voltage from-50mV reached the maximum of+60mV, the Ito was similar between Control 1 and Experiment 1 sub-groups, P>0.05.②When the self-command voltage at+10mV, the current density of ICa,L was slightly higher in Control 2 sub-group than that in Experiment 2 sub-group, P>0.05.
Conclusion: Direct perfusion of cilostazole in right ventricular cells may inhibit Ito in experimental rats, such effect was similar with cilostazole treatment at (1-50)μmol/L. Cilostazole might prevent the ventricular arrhythmia in Brugada syndrome in experimental rats.

Objective To investigate the effects of RNA interference (RNAi) targeting angiotensinconverting enzyme (ACE) on blood glucose,insulin resistance,as well as oxidative stress in type 2 diabetic rats.Methods Type 2 diabetic rats were divided into diabetes control group (caudal intravenation with control adenovirus named Ad5),gene treatment group (caudal intravenation with recombinant adenoviral vectors named Ad5-ACE-shRNA,expressing ACE gene-specific shRNA),and enalapril group (intragastric administration with enalapril every day).At the same time,the normal blood glucose control group was set up.All rats were injected two times during the experiment period.Blood glucose was measured before and after the intervention.At the third day of the experiment,expressions of ACE mRNA and protein in pancreas were evaluated by RT-PCR and Western blot,and serum concentrations of ACE and Ang Ⅱ were measured by ELISA.By the end of the experiment,insulin sensitivity index was calculated and expressions of glucose transporter 4 (GLUT4) protein of epididymal adipose tissue and NAD (P) H (p22phox) protein of pancreas were measured.Results Blood glucose levels in the gene treatment group [(17.8 ±1.1) mmol/L] and the enalapril group [(17.9 ± 1.2) mmol/L] were lower than that in the diabetes control group [(24.9 ± 1.3) mmol/L] when the experiment was finished.ACE mRNA and protein expressions in pancreas of the gene treatment group were significantly decreased compared with the diabetes control group (P ＜ 0.05).Serum concentrations of ACE and Ang Ⅱ in the gene treatment group were (16.37 ± 3.01) ng/ml and (18.24 ± 3.69)pg/ml,significantly lower than those of the diabetes control group [(46.67 ± 3.92) ng/ml and (44.93 ± 4.12) pg/ml respectively,both P＜0.05].Insulin sensitivity indexes of the gene treatment group and the enalapril group were (-5.14 ± 0.41) and (-5.17 ± 0.38),being all significantly higher than that of the diabetes control group (-6.18 ±0.46,both P＜0.05).Expressions of GLUT4 protein in epididymal adipose tissue were higher and expressions of p22phox protein in pancreas were lower in the gene treatment group and the enalapril group than those of the diabetes control group (both P＜0.05).Conclusions RNAi targeting ACE gene may delay the progress of hyperglycaemia and improve the situation of insulin resistance and oxidative stress.The RNAi technology may be used as a new strategy of gene therapy for diabetes mellitus.

Objective:To investigate the influence of social support to nursing students occupation maturity, provide a new perspective and reasonable suggestions for the cultivation of nursing students occupation planning. Methods:A total of 759 nursing students of different education levels were selected. The Social Support Rating Scale and the Career Maturity Inventory Attitude Scale were used to investigate the influence of social support to nursing students occupation maturity. Results:Different levels of nursing students in subjective support and utiliza-tion of support,the total score difference was statistically significant(P <0. 05),college students social support score lower than the college students. Overall maturity of undergraduate and college nursing students occupation de-gree higher than secondary school students,students occupation maturity in the condition assessment,personal ad-justment,value conception,occupation cognition,self-cognition of these dimensions also exist a significant differ-ence(P<0. 05). Social support and occupation maturity of each factor is correlated in most dimensions. Conclu-sion:The social support of nursing students′ occupation maturity degree has important influence,requirements of professional teachers should integrate education occupation maturity while imparting professional knowledge,lay a solid foundation for nursing students′occupation career planning.

Objective To investigate the potential role of MCP-1/CCL2 in experimental acute necrotizing pancreatitis (ANP) and complications. Methods 60 SD male rats were randomly divided into 3 groups: sham operation group ( n = 20 ), ANP group ( n = 20 ) and MCP-1 group ( n = 20 ). ANP model was induced by retrograde infusion of 3.5% sodium taurocholate, MCP-1 group received subcutaneous injection of MCP-1 antibody 0 h and 6 h after ANP induction. The serum levels of amylase, MCP-1, D-lactic acid,histological changes and the expression of MCP-1 mRNA of lung, small intestine and pancreas, the expression of MCP-1 protein in pancreas, MPO levels of small intestine MPO were determined. Results The serum levels of amylase, MCP-1, D-lactic acid in MCP-1 group at 12 h were (4666 ±412)U/L, (39.53 ±8.25)pg/ml and (6.3 ±2.2)mg/L, which were significantly lower than those in ANP group [ (9611 ±363)U/L, (63.42 ±9.32) pg/ml, (9.3 ± 2. 1 ) mg/L, P< 0.05 ) ]; the expression of MCP-1 mRNA in pancreas, small intestine and lung were 0.431 ± 0.009, 0. 211 ± 0.018 and 0.442 ± 0.017, which were significantly lower than those in ANP group [ (0.624 ±0. 010, 0. 523 ±0. 019 and 0. 569 ±0. 024, P <0.05) ]; the expression of MCP-1 protein in pancreas was 2.0 ± 0. 1, which was significantly lower than that in ANP group (4. 0 ± 0. 2, P <0.05). Lung and small intestine MPO were (11.1 ±3.0)U/g and ( 19.2 ±2.0)U/g, which were significantly lower than those in ANP group[(39.2±3.1)U/g and(13.1±2.1)U/g, P<0.05]. Conclusions Early blockade of MCP-1 not only attenuates the severity of ANP, but also decreases the degree of acute lung injury and intestine barrier dysfunction.

BACKGROUND:The mechanism and effect of glycogen synthase kinase 3β(GSK-3β) in the differentiation of cardiac stem cel s into cardiomyocytes are stil unclear, although GSK-3βis closely related to the life activities of cel s.
OBJECTIVE:To investigate the changes of GSK-3βexpression in the treatment of myocardial infarction in rats undergoing cardiac stem cel transplantation.
METHODS:The isolation and culture of cardiac stem cel s were performed in 10 neonatal rats. Lentivirus overexpressing GSK-3βor LacZ (control) was constructed and transferred into cardiac stem cel s. Animal model of myocardial infarction was made in 30 Sprague-Dawley rats. Six weeks after model preparation, rat models were assigned into GSK-3β, LacZ or PBS group. GSK-3βor LacZ overexpressing cardiac stem cel solution or PBS in equal volume was injected into the rat myocardium, respectively. Four weeks after transplantation, the cardiac function and myocardial col agen production in rats were detected and compared.
RESULTS AND CONCLUSION:Compared with the other two groups, the left ventricular ejection fraction was significantly higher, and the left ventricular end diastolic diameter was significantly lower in the GSK-3βgroup (P<0.05). Hydroxyproline content, type I col agen mRNA, and type III col agen mRNA expression were significantly lower in the GSK-3βgroup than the other two groups (P<0.05). Findings from Masson staining showed that the content of blue-stained col agen was significantly lower in the GSK-3βgroup than the LacZ group. Moreover, lowest myocardial infarction size was found in the GSK-3βgroup (P<0.05). Al these experimental findings show that GSK-3 overexpression plays a positive role in promoting the therapeutic effect of cardiac stem cel transplantation.

Objective To investigate the effect of Yiqi Huoxue Huatan Decoction(YHHD) on gene expression of vascular endothelial growth factor(VEGF), intercellular adhesion molecule 1(ICAM-1) in the colonic mucosa of ulcerative colitis rats. Methods Sixty rats were divided into six groups, namely normal control group, model control group, low-, middle- and high-dose Y HHD group(gastric gavage of YHHD in the dosage of 13, 26, 52 g·kg-1·d-1 respectively) , and Mesalazine(0.5 g·kg-1·d-1 ) group. T he rat model of ulcerative colitis was established by enema with 2, 4, 6-trinitrobenzene sulfonic acid(TNBS)mixture(2.5%TNBS and 50% ethanol). The changes in rat body weight were monitored. After treatment, pathological features of the colonic mucosa were observed under light microscope, and VEGF and ICAM-1 gene expression levels in the colonic mucosa were measured by reverse t ranscripti on polymerase chai n reaction(RT-PCR). Results Body weight was lighter, and relative gene expression levels of VEGF and ICAM-1 in the model group were significantly higher than those in the normal control group, and pathological features of the colonic mucosa was obvious, the differences being significant between the two groups(P<0.01). Compared with the model group, body weight was increased , pathological features of the colonic mucosa were relieved, and relative gene expression levels of VEGF and ICAM-1 were significantly decreased in YHHD groups(P<0.01), and high-dose YHHD showed the strongest effect. Conclusion YHHD can down-regulate the gene expression of VEGF and ICAM-1, which may be one of its therapeutic mechanisms for ulcerative colitis.

Objective To explore the clinical features of lissencephaly and the detection ofLISI gene.MethodsThe characteristic of clinical features, laboratory examination and gene detection in one case of lissencephaly was retrospectively analyzed. Meanwhile, the related literatures were reviewed.ResultsA 5-month-old female child diagnosed with epilepsy 20 days ago was hospitalized for convulsive seizure more than 30 times in 3 days. The manifestations were eyes staring, and turning upward, cyanosis of lips and face, froth at the mouth, extremities rigidity and loss of consciousness, and the symptoms can spontaneously remitted in 2-3 minutes. Laboratory examination showed that peripheral blood white cell count was 13.67×109/L, hemoglobin 108 g/L, red blood cell count 3.90×1012/L, lymphocyte 10.26×109/L; maocardial enzyme and hepatic and renal function were normal; blood ammonia was 23 μmol/L and lactic acid 2.11 mmol/L. Long-range video EEG showed highly arrhythmia, and frequent partial epilepsy, and sometimes secondary generalized epilepsy. Head MRI showed lissencephaly. The child was treated with oral administration of Keppra 27 mg/(kg·day), Topiramate 6.5 mg/(kg·day), currently no seizure. The detection ofLIS1 gene found that heterozygous mutation of c.232delG, which lead to protein shift mutation (p.E78NfsX25). No mutation was found in her parents.ConclusionsChild with lissencephaly may combine with epilepsy which may cause by mutation inLIS1 gene. And there was no information about point mutation of c.232delG inLIS1 gene being reported at home and abroad so far.