BACKGROUND:Caveolin-1 is expressed in mammalian brain and involved in the normal development of the brain, which can affect the proliferation of neural stem cells in the brain. OBJECTIVE:To acquire neural stem cells from caveolin-1 knockout embryonic mice in vitro and study their biological characteristics. METHODS:The whole brain was separated from C57BL/6 mice and caveolin-1 knockout C57BL/6 mice respectively at encyesis 14-16 days. Single cellsuspension was obtained by enzyme digestion, and cultured in the conditioned medium of neural stem cells. Fol owing 7 days of primary culture, the cells were induced in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum for 7 days. RESULTS AND CONCLUSION:The major cells of the cellsuspensions from the fetal mouse brain were dead at 1 day after culture, and some single cells floated in the medium and their transmittance were better, and then they gradual y formed multicellular bal s after 3 days. A smal amount of cells were adhered at the bottom of culture plate after passage, and a great amount of cellbal s appeared after 7 days. The proliferation rate of neural stem cells from caveolin-1 knockout mice was higher than that from normal mice. The cellbal s were nestin-positive and their differentiated cells was positive for neurofilament 200, glial fibril ary acidic protein or O4, respectively. Al of the cells from normal mouse brain were positive for caveolin-1, but the cells from caveolin-1 knockout mice were negative for caveolin-1 by immunocytochemistry. Moreover, the speed of cellbal formation and the number of cellbal s in neural stem cells from caveolin-1 knockout mice were better than those from normal mice. Caveolin-1 negative neural stem cells were cultured successful y from caveolin-1 knockout mouse brain, and the results show that caveolin-1 can promote the proliferation of neural stem cells and inhibit their differentiation in vitro.

AIM: To explore the mechanism of myocardium protection after ischemia/reperfusion (I/R) injury by preconditioning with ischemia in human. METHODS: Thirty - six patients underwent valve replacement were divided into ischemic preconditioning group (IP group, 20 cases) and non -ischemic preconditioning group (control group, 16 cases) according to whether they were given single cycle reperfusion before cardioplegia or not. Serum levels of interleukin -8 and 10 were measured with ELISA. Expressions of myocardial Bel -2 and caspase -3 were analyzed. RESULTS: The inflammatory factors IL - 8 and IL - 10 increased to the highest level in serum at 6 h after declamping and recovered to normal level on 5 d after declamping. On 6 h, 1 d and 2 d after declamping, serum level of IL -8 was significantly lower in IP group than that in control group ( P < 0.05 ) , but serum level of IL -10 was higher in IP group (P < 0.05 ). Expression of myocardial Bel - 2 and caspase - 3 increased in both groups after reperfusion, and Bel - 2 was lower in the control group than that in IP group while the level of caspase - 3 was higher (P < 0.05). Expression of myocardial Bel - 2 had positive correlation with IL - 10 and negative correlation with IL - 8.CONCLUSION: Ischemic preconditioning has the effect of protection of human myocardial cells after ischemia/reperfusion injury through decreasing systemic inflammatory response following ischemia reperfusion injury.

Objective To explore the effect of Buyang Huanwu Decoction (BYHWD) on vascular endothelial growth factor (VEGF) and its receptor, Fetal liver kinase 1 (Flk1) in rats after focal cerebral ischemia. MethodsRats were randomly divided into following groups including Sham group, model group, Nimodipine group, and BYHWD group. The model of focal cerebral ischemia in rats was repro-duced by middle cerebral artery occlusion. Rats were ig administered and killed after operated, then the expression of VEGF and Flk1 and protein level of VEGF in brain tissue of every groups were measured by immunohistochemisty and enzyme linked immunosorbent assay, and the nervous function deficit scores were evaluated. ResultsThere were a few VEGF and Flk1 positive cells in normal brain tissue of rats. After focal cerebral ischemia the VEGF and Flk1 positive cells were increased. Compared to model group, Nimodipine and BYHWD significantly improved the neurological behavior performance, increased the numbers of VEGF and Flk1 positive cells, and enhanced the VEGF protein level (P

Objective To explore the clinical effect of endoscope emergency operation in the treatment of gastric variceal bleeding.Methods According to the digital table,90 patients with acute bleeding gastric varices were randomly divided into the control group and the observation group,each group 45 cases.The control group only received drug treatment,and the observation group was given endoscopic therapy.The clinical effect of the two groups were compared.Results The immediate hemostatic rate,rebleeding rate,varicose veins subsided total efficiency of the observation group were 100.00％,17.78％,86.67％,respectively,which were better than those of the control group (84.44％,44.44％,28.89％) (x2 ＝ 5.72,P ＜ 0.05).Conclusion Endoscope emergency operation combined with oral drug in treatment of gastric variceal bleeding has good effect,can achieve rapid hemostasis,prevent recurrence,and it is worthy of clinical application.

Objective To explore effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide(VIP) in the hypothalamus, spinal cord and duodenum of chronical stressed mice. Methods Mice were randomly divided into normal, stress and Simotang group( n= 10 in each group), and given a variety of unpredictable chronic mild stress. After 21 days gastric emptying and intestinal propulsion function were measured,the expression of VIP was detected by immunohistochemistry and RT-PCR. Results Compared with mice in normal group( (49.81 ± 8.56)%; (51.02 ± 5.11 )% ), chronic stress increased gastric residual rate( (61.53 ±8.71 ) %; P ＜ 0.05 ) and reduced small intestine propulsion rate ( ( 31.79 ± 2.38 ) %; P ＜ 0. 05 ). There were differences in expressions of VIP positive cells and mRNA in duodenum( (8.8 ± 1.1 )/mm2 and(0. 58 ±0.03) ),hypothalamus ( ( 12.9 ± 1.5 )/mm2 and (0.81 ± 0. 07 ) ) and spinal cord ( ( 12.1 ± 1. 2)/mm2 and (0.76 ± 0.02) )in chronic stress group compared with normal group( (6.5 ± 0. 9)/mm2 and (0.43 ± 0. 04);( 10.8 ± 1.3 )/mm2and (0.57 ± 0.03 ); (9.3 ± 1.5 )/mm2 and (0.53 ± 0. 02 ) respectively). There was not difference in gastric residual rate (52.93 ± 9.15 )%, small intestine rate(48.98 ± 4.38 )% and expressions of VIP positive cells and mRNA in duodenum ( (6.7 ± 0.9)/mm2 and (0.48 ± 0. 05 ) ), hypothalamus ( ( 10. 6 ± 1.4 )/mm2 and ( 0. 61 ± 0. 05 ) )and spinal cord ( (9. 1 ± 1.3)/mm2 and(0.55 ± 0.05 ) ) in Simotang group compared with those in normal group (P ＞ 0.05 ), but there were decreased compared with those in chronic stress group (P ＜ 0.05 ). Conclusion Simotang can regulate expressions of VIP in duodenum, hypothalamusand spinal cord in chronically stressed mice.

Objective To summarize the preservation measures of the donor's heart and lung, and the postoperative immunotherapy, as well as the clinical experience of discrimination and management for graft rejection.Methods The clinical data of 2 cases of heart-lung transplantation in our department were retrospectively analyzed. Two different protective liquids were used for donor's lung lavage of 2 cases: Perfadx solution (1000 mL containing tris 0.3 mL and ilomedin 25 μg); Euro Collins solution (1000 mL containing tris 0.3 mL and PGE1 100 μg). UW solution was used for donor's heart lavage. Surgical procedure for heart-lung transplantation was classic technique in situ. The schedule of immunosuppression was induced by Basiliximab, and combined with cyclosporine+ mycophemolate mofeil+corcal hommone after operation. recipient's blood count, organ's functions, the sizes of every cavity of heart, IVSPW and LVPW were observed during early post-operation. The recipients were subjected to chest CT scan, fiberoptic bronchoscope and tissue pathological study when necessary to find the signs of rejection promptly. When the rejection occurred in the recipient, cortical hormone's impulse therapy was given and the dose of immunosuppression was adjusted in time.Results Two patients discharged in 80 days and 141 days after operation. The patients were followed up for 54 months and 50 months respectively, and their life qualities were very well. Acute rejections occurred on the 10th and 26th day in one case, and in another case, acute rejections occurred on the 29th and 87th day after operation. All were conversed by cortical hormone's impulse therapy and adjusting the dose of immunosuppressants. When acute rejection occurred, the blood count had significant change, and IVSPW and LVPW were increases. They were returned the normal range after corresponding therapy.Conclusion Perfidx solution and Euro-Collin solution may play good protective roles for donor's lungs. UW solution may play good a protective role for donor's heart. To discriminate the clinical graft rejection and infection in time and administrate correct management will have large benefits for the patients' rehabilitation.

Objective To study the feasibility and safety of limb remote ischemic preconditioning (RIPC) in infants and explore the protective effect on myecardium ischemia reperfusion injury for infants undergoing cardiac operation under cardiopulmonary bypass. Methods 60 infants weight less than 7 kilograms with ventricular septal defect were enrolled into the study. 30 of them (RIPC group) were ischemic preconditioned two times (24 hours and 1 hour preoperatively) by three cycles of iscbemia (5 minutes for each) and reperfusion on the left upper arm using a blood pressure cuff. Serum lactate dehydrogenase (LDH), creatine kinase (CK) and its isoenzyme (CK-MB), and tro-ponin I (TnI) ; malondialdehyde (MDA) and superoxide dismutase (SOD) was preoperatively detected. The expression of heat shock pro-tein 70 (HSP 70) in cardiomyocytes was determined by western blot analysis. The surgical outcome including limb movement and sensory function was also recorded. Results No limb disability or sensory disturbance or no other surgical complications was found in all infants. LDH, CK, TnI at the beginning of operation in RIPC group was higher than those in control group. After operation, leakage of heart enzymes were attenuated in RIPC group, and the serum concentration of enzymes were lower than those in the control group. The RIPC group had low coronary sinus venous concentration of MDA but high SOD. The expression of HSP70 was upregulated in cardiomyocytes of RIPC group. Conclusion The limb RIPC can be done easily and safety in infants, and BIPC can reduce the leakage of myocardial enzymes and upregu-late the expression of HSP, which possess protective effect on myocardial IRI.

Objective To investigate the effects of L-carnitine on myocardial enzymes in infants with congenital heart disease undergoing open cardiac operation with cardio pulmonary bypass (CPB).Methods From Mar.2011 to Nov.2012,there were 60 infants with ventricular septal defects,whom were divided randomly into the test (n =30) and control (n =30) groups.L-carnitine was put in the cold crystal cardiac arresting liquid in the test group (6 g/L),other experimental conditions were the same between two groups.Before CPB,at 1 h,24 h,7 d after the pump-off,venous blood was drawn to test the level of serum creatine kinase (CK),CK-MB isozyme,cardiac Troponin I (cTnI).Blood samples were taken at different time points for the analysis of tlymphocyte and its subtype by flow cytometry.Observing the dopamine and other vasoactive drug 8 hours and 24 hours after the pump-off,using data obtained from the t-test done statistically.Results The levels of CK and CK-MB and cTnI had no difference between two groups before operation.From the end of CPB,the levels of CK and CK-MB and cTnI were significantly lower in the test group than that in the control group (P ＜0.05).The automatic jump higher rate (95％ vs 70％,P ＜ 0.05) and the amount of dopamine were significantly less than the control group (P ＜ 0.05).Conclusions L-carnitine improved obviously microcirculation of ischemic cardiac reperfusion injury undergoing open cardiac operation under CPB and heart function.It had a good protective effect on myocardium in infants.

AIM:To observe the expression of hypoxia-inducible factor 1 (HIF-1) and neuroglobin (NGB) in piglet cortex during deep hypothermic circulatory arrest.METHODS:Wuzhishan piglets were randomly assigned to car-diopulmonary bypass group ( CPB group) , 40 min of circulatory arrest ( CA) at 18 ℃ without cerebral perfusion ( DHCA group) or with selective antegrade cerebral perfusion ( SACP group) .After 180 min of reperfusion, cortical tissue was har-vested for determining HIF-1αand NGB expression by HE staining, Western blot and real-time PCR.RESULTS:Severer cerebral injury was observed in DHCA group than that in SACP group.After 180 min of reperfusion, HIF-1αprotein and mRNA levels were significantly higher in DHCA group than those in CPB group (P<0.05).Accordingly, SACP animal had higher levels of HIF-1αprotein and mRNA than those in DHCA group (P<0.05).Simultaneously, higher NGB pro-tein and mRNA levels were found in DHCA group than those in CPB group after 180 min of reperfusion ( P<0.05) .The SACP animal had higher levels of NGB protein and mRNA than those in DHCA group (P<0.05).CONCLUSION:Up-regulation of HIF-1 and NGB are involved in the mechanism against cerebral injury resulting from DHCA in the cortex and possibly a part of cerebral protective effect of SACP.

Objective To investigate the changes and significance of serum anti-CCP (ACCP)antibody,matrix metalloprotein-ase-3 (MMP-3 ),interleukin-17 (IL-17 ),interleukin-18 (IL-18 )in the patients with rheumatoid arthritis (RA).Methods The ELISA method was adopted to detect the in the peripheral blood serum ACCP antibody,MMP-3,IL-17 and IL-18 in 80 patients with RA,32 patients with osteoarthritis (OA)and 32 cases of healthy controls and the detection results were performed the statis-tical analysis.Results The ACCP antibody,MMP-3,IL-17 and IL-18 levels in the RA group were significantly higher than those in the OA group and the healthy control groups;the ACCP antibody,MMP-3,IL-17 and IL-18 levels in the low,middle and high RA activity groups were higher than those in the stable group,the ACCP antibody,MMP-3,IL-17,IL-18,disease activity score (DAS28),C-reactive protein (CRP)and erythrocyte sedimentation rate(ESR)in the RA group were increased;the MMP-3,IL-17, IL-18 and DAS28 in the ACCP antibody positive group were higher than those in the A CCP antibody negative group;the positive correlation existed among the ACCP antibody,MMP-3,IL-17 and IL-18 in the RA group (P <0.05);the ACCP antibody,MMP-3, IL-17 and IL-18 were positively correlated with the monitoring indicators of CRP and DAS28 in the low,middle and high RA activi-ty groups.Conclusion MMP-3,IL-17 and IL-18 participate in the occurrence and development process of RA;The detection of ser-um ACCP antibody,MMP-3,IL-17 and IL-18 has a certain value in the judgment of disease activity,and prevention and treatment in the patients with RA.