Objective To construct lentivirus vectors carrying alphastatin gene, test its secretion expression in human umbilical vein endothelia cells (HUVECs) and observe its effects on growth, migration and tube formation of HUVECs. Methods We constructed recombinant lentivirus vectors of NT4-alphastatin fusion gene containing neurotrophin-4 signal peptide, pro-region sequences and alphastatin, then transfected the recombinant lentivirus vectors into HUVECs to obtain secretory protein alphastatin and test its anti-angiogenic activities in vitro. Results Our data showed that recombinant self-inactivating lentivirus vectors of NT4-alphastatin were successfully constructed, and stable NT4-alphastatin transduced HUVECs were capable of sustainably secreting alphastatin which significantly suppressed HUVECs migration and differentiation but not VEGF-induced proliferation. Conclusion This report represents the first time on the use of lentivirus-based vectors to deliver alphastatin, the endogenous angiogenesis inhibitor, and reveals the potential utility of anti-angiogenic gene therapy with lentivirus vectors for treating cancer.

Objective To investigate the protective effects of morphine preconditioning on the lungs against ischemia-reperfusion (I/R) injury and the possible mechanisms.Methods Twenty-four Japanese long-ear white rabbits weighting 2.5-3.0 kg were used in this study. The animals were anesthetized with intramuscular atropine 0.5 mg and intravenous 3 % pentobarbital 30 mg ?kg-1, tracheostomized and mechanically ventilated ( VT 10 ml?kg-1, RR 30 bpm, FiO2 100% , PEEP 1 cm H2O) . Right carotid artery was cannulated for BP monitoring and blood sampling. An elastic band was placed around the hilum of left lung via thoracotomy to perform lung ischemia. Body temperature was maintained at 36-38℃( rectal) . The animals were randomly allocated to one of 3 groups (n = 8 each) : (1) Sham group; (2) I/R group was subjected to 2 h in situ left hilar occlusion followed by 2 h reperfusion and (3) morphine preconditioning group received morphine 4 mg?kg-1 via pulmonary artery 30 min before I/R. Mean pulmonary artery pressure (MPAP) and peak inspiratory pressure (PIP) were monitored and recorded. Arterial blood samples were taken before occlusion of lung hilum (baseline) and at 5, 30, 60, 90 and 120 min of reperfusion for blood gas analysis, and determination of plasma endothelin-1 concentration. The animals were killed at the end of 120 min reperfusion. The lungs were removed for determination of lung water content (W/ D ratio), percentage of neutrophils in the broncho-alveolar lavage fluid ( BALF) and microscopic examination.Results MPAP and PIP were significantly lower while PaO2 was significantly higher at 30, 60, 90 and 120 min of reperfusion in morphine preconditioning group than in I/R group. Plasma endothelin-1 concentration was significantly lower at 60 and 120 min of reperfusion in morphine preconditioning group than in I/R group ( P

Objective: Effects of surfactant replacement were evaluated on graft pulmonary function in rats. Method: Forty adult Wistar rats were anesthetized with an intraperitoneal injection of pentobarbital sodium(30 mg?kg~(-1)), and intubated through tracheolomy. All rats were ventilated with a ventilator. Surfactant of 50 mg was injected into lungs through tracheal. Cold Ringer's solution(0℃-4℃)was infused into the lungs through pulmonary artery, and then they were kept in cold solution. Result: The static lung compliance (SLC)in the both surfactant-replaced S(no airway pressurc)and SP(0.98kPa airway pressure)groups did not markedly decrease after cold storage. However,SLC in the no surfactant-replaced NP (no airway pressure)and P (0.98kPa airway pressure) groups both significantly decreased after cold storage. SLC was higher after lh in the P group than NP group(P

0.05). After the injection, PaO 2 in group A, B and C increased significantly, whereas PaO 2 in group D did not change and was kept at low level below 13.3kPa (P

ObjectiveTo investigate the effect of therapeutic hypercapnia on hepatic ischemia-reperfusion (I/R) injury in rat liver transplantation.MethodsMale specific pathogen-free adult Wistar rats aged 6 weeks weighing 220-280 g were used in this study.Sixteen rats in which liver transplantation was successfully performed were randomly divided into 2 groups ( n =8 each): liver transplantation group (group LT) and therapeutic hypercapnia group (group TH).In group TH,PaCO2 was maintained at 80-100 mm Hg by inhalation of CO2 for 1 h at the begining of reperfusion.MAP,PaO2 and PaCO2 was recorded during reperfusion.Blood samples were obtained at 2 h of reperfusion for determination of serum ALT,AST,TNF-α,IL-1 and IL-6 levels,and then the rats were sacrificed and transplanted liver was immediately removed for determination of NF-κB activity and apoptosis and microscopic examination.The apoptotic index was calculated.ResultsMAP,PaO2 and PaCO2 were higher,and serum ALT,AST,TNF-α,IL-1 and IL-6 levels,NF-κB activity and apoptotic index lower in group TH than in group LT ( P ＜ 0.05).The histopathologic damage was ameliorated in group TH as compared with group LT.Conclusion Therapeutic hypercapnia can attenuate hepatic I/R injury in rat liver transplantation by inhibiting inflammatory response and apoptosis.

Objective To summarize the experience in diagnosis and treatment for solid-pseudopapillary tumors of the pancreas (SPT). Methods A retrospective clinical analysis about clinical, imaging and pathologic data was made on 16 cases of SPT admitted from January 2005 to December 2009. Results Five had SPT in the head of the pancreas, 5 in the body of the pancreas, 6 in the tail of the pancreas. The first symptom was intermittent epigastric pain ( n = 7), abdominal aponia mass ( n = 3), Pancreatic tumor found by chance (n =4), weight loss (n =2). Solid and Solid-cystic masses of low echo were found in US. Masses of low density in pancreas were found on CT scan, while irregular enhancement appeared in the circumference of all tumors in enhanced CT scan sequences. Tumor markers in patients' erum were all negative.9 patients underwent distal pancreatectomy and spleen resection, including 1 patient also underwent left hemicolectomy. Local excision of tumor was performed in 4 cases. Pancreatic local excision and pancreaticojejunostomy were performed in 3 cases. 14 cases were followed up with an period of from 3 to 48 months. No evidence of relapses and metastasis in these cases was found. Conclusion SPT primarily affects young women, and it may be located in any part of pancreas. Surgical resection is recommended as the treatment of choice. The prognosis is good.

Objective To investigate the role of oridonin in preventing skin graft rejection.Methods BALB/c mice were transplanted with skin grafts from C57BL/6 mice.Grafted mice were treated daily with oridonin,CsA and PBS,respectively.The survival of grafts was inspected daily and evaluated by histological analysis.On day 7 after transplantation,the percentage of CD4+ CD25+ Foxp3+ cells (Treg) in the spleen was determined by flow cytometry.The effect of oridonin on MLR and apoptosis was examined in vitro.Naive BALB/c mice were intraperitonealy injected with oridonin (15 mg/kg/day).At different time points,the number of T cells and macrophages in peripheral blood mononuclear cells (PBMCs) as well as the spleen was examined.Results The survival of skin grafts in the oridonin group (15.8 ± 1.5 days) was significantly longer than that in the control group (12.3 ± 1.2 days) and the CsA group (13.3 ± 1.1 days).Oridonin reduced inflammatory cell infiltration in grafts.The expression of Tregs was higher in the oridonin group (17.6 ± 3.6％) than in the control group (14.8 ± 2.3％).In vitro oridonin inhibited MLR and induced apoptosis in a dose-dependent manner.The number of T cells in PBMCs was rapidly decreased following oridonin treatment.With the depletion of T cells in PBMCs,high frequency of granulocytes was observed.On day 8,the number of T cells in the spleen was decreased,which was accompanied by increased phagocyte number.Conclusion Oridonin could suppress allograft rejection and prolong survival of skin grafts.The mechanism may be attributed to upregulation of Tregs and clearance of T cells.

Objective To investigate the effects of N-acetylcysteine (NAC) pretreatment on the liver function and mRNA and protein expressions of nuclear factor-KB (NF-KB) in brain-dead BA-Ma mini pigs. Methods The brain-dead model was established by increasing intracranial pressure by a modi-fied, slow and intermittent way. A total of 15 BA-Ma mini pigs were randomly and equally divided into three groups (five in each group), ie, control group (Group C) : treated only with opening and closing abdomen after anesthesia; group without NAC treatment (group B): brain-dead models without use of NAC; NAC treatment group (Group N): 1 and 12 hours after establishment of brain-dead models, 200 mg/kg NAC was added into 100 ml normal saline and intravenously transfused. Levels of ALT and AST in serum as well as TNF-α, IL-1β and IL-6 were determined at 3,6,12, 18,24 hours after brain death. The changes of liver tissues were observed by HE staining under a light microscope, the uhrastruc-rural changes of liver tissues observed under electron microscope, the expression of NF-KB detected by immnohistochemistry and change of NF-KB mRNA by RT-PCR. Results (1) Compared with Group C, serum ALT and AST began to increase at 12 hours after brain death, but IL-1β, IL-6 and TNF-α be-gan to increase three hours after brain death in Groups B and N. mRNA and protein expressions of NF-KB in Groups B and N began to increase six hours after brain death, when Group B increased more sharply than Group N, with statistical difference (P<0.05). (2) At 12 hours after brain death, injury of liver cells in Group B was severer than that in Group N. Conclusion NAC can inhibit the mRNA and pro-tein expressions of NF-KB, decrease the release of inflammatory factors and hence protect the hepatic structure and function during brain death.

Objective To study the efficacy and the safe dosage of high intensity focused ultrasound (HIFU) ablation in pancreatic cancer.Methods From November 2010 to May 2013,21 patients with advanced pancreatic cancer were treated by HIFU at the First Affiliated Hospital of Zhengzhou University.These patients who were randomly divided into two groups (10 and 11 patients respectively),were given a low-power (100～249 W) treatment and a high power (250 ～350 W) treatment.These two groups of 21 patients received a total of 25 times of HIFU treatment (3 patients received twice of low-power treatment,while 1 patient received twice of high-power treatment).The two groups were compared by analyzing the treatment parameters (average power,total treatment time,treatment total energy,treatment volume,etc.) and volume of tumour response as shown on postoperative imaging (CT or MRI) examinations.Also,the complications,degree of pain relief and survival were compared.The energy efficiency factor (EEF) and the ablation ratio were calculated.A preliminary study was conducted on the relationship of the ultrasound dose and the ablation effect of HIFU treatment for pancreatic cancer.Results (1) The EEF of the high-power group (≥250 W) and the low-power group (＜ 250 W) were (10.39 ± 5.71) J/mm3 and (21.62 ± 9.81) J/mm3,the former group was significantly lower than the latter group (P ＜0.05) ; the ablation ratio of the high-power group was higher than the low-power group,(91.52 ± 4.18)％ versus (51.59 ± 7.66)％ respectively,the difference was statistically significant (P ＜ 0.001).(2) The efficiency factor and the ablation volume for the HIFU treatment showed a linear trend,and both were negatively correlated (Pearson correlation coefficient r =-0.485,P ＜ 0.05).(3) There was no serious complication after the HIFU treatment.In the low-power group,six of ten patients were alleviated of his pain (60％) ; the CA19-9 decreased in four of ten patients after HIFU treatment (40％).In the high-power group,nine of eleven patients were significantly relieved of pain after treatment (82％),the CA19-9 decreased in five of nine patients after HIFU treatment (56％).(4) On Kaplan-Meier survival analysis,HIFU treatment of patients with pancreatic cancer,the median survival was 8 months and 9 months in the low-power group and high power group,respectively (Log-rank test x2 =0.05,P =0.944).Conclusion During HIFU treatment of patients with pancreatic cancer,if the ultrasound power was between 250 W and 350 W,there was a higher proportion of tumor ablation,but with no serious complications.Thus,this dose was safe.

Objective To explore the relationship between the hepatic injury and the severity of the patients with non-gallstone severe acute pancreatitis,so as to provide some evidences in the treatment on the hepatic injury,to lower the systemic and local applications of SAP,as well as lower the morality of SAP.Methods Patients suffering from non-gallstone SAP were retrospectively analyzed.We statisticated the occurrence of multiple organ dysfunction syndrome.We adopted the occurrence of MODS of the patients as the indicatrix of prognosis and analyzed the results.Univariate analysis and multivariate logistic regression analysis were used to analyze the ralationship between the markers of heapatic injury and the prognosis of the patients suffered of non-gallstone SAP.Results The univariate logistic regression analysis of the relationship between hepatic dysfunction and the occur of MODS of non-gallstone SAP shows that there were correlation between the occur of MODS and AST,ALT,ALB,respectively.The multivariate logistic regression analysis shows that there were correlation between the occur of MODS and AST,ALT,ALB,respectively.Conclusion The study showed that the heighten of the serum consentration of AST,ALT,TB and the degrade of the serum consentration of Alb were the independent influencing factors of patients with non-gallstone SAP,respectively,and suggested the unfavourable prognosis of the patients.