Objective To investigate the change of lung morphology and the mechanism in the brain-dead state. Methods Ten Ba-Ma mini pigs were randomized into 2 groups: brain-dead group (group B, n=5) and control group (group C, n=5). Brain-dead model was established in group B by increasing intracranial pressure in a modified, slow and intermittent way, and brain-dead state maintained for 24 h by respiration and circulation support. The serum TNF-?, IL-1?, and IL-6 were determined at 3, 6 12, 18 and 24 h after the initial confirmation of brain death. Lung tissues were taken at 24 h. The alterations of lung tissues were observed by HE staining, and the expression of PKC-? detected by immunohistochemistry. PKC-? mRNA at each time point was detected by RT-PCR. The microstructure of hepatic tissues was observed under an electron microscope. Results (1) Under the light microscopy, broadened lung alveolar septum, edematous, extravasate in the alveolar congestive capillary vessel, infiltration of lymphocyte were observed. Under the election microscopy, cytoplasm edema, swollen mitochondria of the type-Ⅱ epithelial cells, partial membrane dissolution of mitochondria were observed and the microtomentum disappeared. No obvious morphological injury was observed under light microscopy and election microscopy. (2) Compared with control group, the serum IL-1?, IL-6 and TNF-? in brain-dead group began to increase at 3 h after brain death, and become more and more higher when the maintained state was prolonged. The expression of PKC-? and PKC-? mRNA in brain-dead group was increased at 24 h in the lung tissue. Conclusion Brain death may evoke lung morphological injury and increase the level of inflammatory mediators; After brain death the levels of PKC-? mRNA transcription and protein translation is increased. The activation of PKC-? and the alteration of inflammatory mediators are supposed to be one of the mechanisms of the lung injury.

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 summarize the diagnosis and therapeutic experience of insulinoma in order to improve the surgical success rate and prognosis. Methods The clinical data of 138 patients with insulinoma from 1966 to 2007 were retrospectively analyzed. Results In this group of patients, hypoglycemia of different levels and Whipple triad were detected. 64 patients expressed different psychic symptom, 12 patients' psychic symptom were still present after blood glucose normalized after operation. Fasting serum insulin values in 88 patients were measured, and the insulin release index was higher than 0. 3. Before operation, tumor was detected in 8 of 75 patients by B-ultrasound scan, and in 17 of 68 patients by CT, and in 5 of 10 patients by MRI. The intra-operative B-ultrasound (IOUS) examination was applied in 44 cases, and 43 cases were successfully detected. The operations included enucleation of insulinoma (n=88) , resection of the body and tail of pancreass (n = 44) , pancreaticoduodenectomy (n=2) , and biopsy (n=1). The blood glucose symptoms normalized postoperatively in 132 patients. The blood glucose rebound in 110 patients, but blood glucose normalized within 2 weeks. After operation, 20 patients developed pancreatic fistula, 32 patients developed acute pancreatitis. Conclusions Insulinoma could be qualitatively diagnosed according to Whipple triad and the insulin release index. Operations with IOUS were simple and effective methods to localize the tumors. The only way to cure insulinoma was operation, and IOUS guided operation could avoid main pancreatic duct and vessel injury, decrease post-operative complications.

ObjectiveTo investigate the mechanisms of neonatal transplantation tolerance,especially the role of immature immune system and chimerism in tolerance.MethodsF1 ( or GFP-F1 ) mice were bred by crossing male C57BL/6 (or GPF transgenic C57BL/6) and female BALB/c mice. Within 24 h,newborn C57BL/6 mice were inoculated with different doses of splenocytes from F1 or GFP-F1 mice,irradiated spleen cells were used as control.Six weeks later,the mice were subjected to F1 skin grafting,and mixed-lymphocyte reaction was performed to determine their tolerance.Flow analysis was used to detect chimerism.ResultsLiving F1 spleen cells could induce chimerism and neonatal transplantation tolerance,but irradiated cells not.The chimerism in long-term tolerant mice is higher than that in chronic rejected mice,with 6.48％ ±4.02％ vs 1.57％ ±0.89％,the difference is significant in statistical analysis.The degree of neonatal transplantation tolerance is determined by the dosage of donor cells,the mice induced with 3 × 107 F1 spleen cells have 80％ long-term tolerance,but the dose of0.7×107 F1 spleen cells could only prolong allografts survival.ConclusionNeonatal transplantation tolerance is dependent on chimerism,the chimerism of donor cells leads to clonal deletion of alloreactive T cells.

Objective To study the methods for diagnosis and treatment of insulinoma.Methods The clinical data of 120 patients with insulinoma who had been admitted to our hospital in the last 40 years were retrospectively reviewed.Results Fasting blood glucose values were less than 2.75 mmol/L in all the patients.Fasting serum insulin values in 75 patients were higher than 25 ?U/mL,and the average was (65 ?6.0)?U/mL.Before operation,tumor was detected in 2 of 60 patients by ultrasound scan,and in 10 of 50 by CT. Among 18 patients who had intra-operative B-ultrasound examination, 16 positive cases were verified by intraoperative exploration; and one case the tumor was not palpable but was found by intraoperative B-ultrasound examination.The operations included enucleation of insulinoma(70 patients),insulinoma resection and distal resection of the pancreas(44),distal resection of the pancreas(4),and biopsy(2).The low blood glucose symptoms disappeared after the first operation in 111 of the 112 patients who had benign tumor.One case with benign tumor was cured by a second operation.Twenty patients developed pancreatic fistula after tumor enacleation, of them,14 healed uneventfully after drainage,5 were cured by operation,and 1 died of peritoneal infection.Conclusions Preoperative localization of insulinomas is difficult. Intraoperative exploration and ultrasound scan are the chief methods for the localization of insulinoma.Enucleation of insulinoma should be selected for benign tumor. Resections of the pancreatic body and tail is required for large,deep or multiple tumors.

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 evaluate the protection effects of glycine on the brain dead donor liver. Methods 42 male Wistar rats were randomized into 3 groups of liver transplantation: brain dead donor (BDD) group (group B), glycine pretreatment group with BDD (group G), strychnine pretreatment group with BDD (group S). For groups B, G and S, the brain death model was established in the donor rats and then liver transplantation was performed utilizing microsurgical techniques. After establishment of brain death state, and during liver cold rinse of donors or liver reperfusion of the recipients, rats in group B were treated with glycine at a dose of 0.6 mmol, 25 ?mol and 25 ?mol in group G, and rats in group S were given the same dose of glycine and strychnine ( 1 000 ∶1), and rats in group B were not treated. Before the cold rinse, at 2 h and 6 h after the portal vein (PV) reperfusion, blood samples were taken from IHVC to determine the levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor alpha (TNF ?) and hyaluronic acid (HA). At 6 h after PV reperfusion, graft samples were fixed for morphological observation and the apoptosis of hepatocytes was detected by using TUNEL method. Results At the time points before liver cold rinse or at 2 h and 6 h after PV reperfusion, serum levels of ALT, AST, TNF ?, HA and apoptosis index (AI) in groups B and S were significantly higher than those in group G ( P 0.05 ). Electron microscopy showed that Kupffer cells were activated and hepatic cells injured more obviously in groups B and S than in group G. Conclusion Glycine may alleviate the injury of the graft from the brain dead rats.

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 investigate the effects of RNA interference targeting EphA7 gene on the growth of SMMC-7721 cell xenograft in nude mice.Methods Recombinant plasmid of EphA7 gene-targeting siRNA was transfected into hepatic cancer SMMC-7721 cells by LipofectamineTM2000,comparing with the empty vector transfected group,untransfected group and control group.The nude mice tumor model was established by subcutaneous injection of hepatic cancer cells in the left upper limb of the mice.Control group was injected with PBS as blank.Real-time PCR,immunohistochemistry and Western blot were employed to detect the mRNA and protein expressions of EphA7 in tumor tissues.The tumor formation time,tumor mass and weight of tumor were also considered in the analysis.Results About 9 ～ 12 days after the injection of tumor cells,the xenograft tumor formation can be observed around the injection site except the control group.35 days after tumor formation,there were obvious decreases in the tumor growth rate,tumor mass,as well as tumor weight in transfected group,comparing with empty vector transfected group and untransfected group (P ＜0.05).Transfection of RNA interference can inhibit the growth of xenograft tumor by 55％.Immunohistochemistry tests showed that there were less cells with positive staining of EPHA7 protein in transfected group,and the staining was lighter as pale yellow,in contrast with the untransfected group and the empty vector transfected group.Real-time PCR and Western blot revealed that the expression of EphA7 mRNA and EPHA7 protein of transfected group were significantly lower than those of untransfected group and empty vector trausfected group with statistically significance (P ＜ 0.05).Conclusion Silencing EphA7 gene with RNA interference can effectively inhibit the growth of SMMC-7721 cell in nude mice,which is expected to become a new target for gene therapy of hepatic cancer.