The rat islets were isolated by using collagenase digestive method, purified by Fi-coll density gradient centrifugation, and rat hepatocytes isolated simultaneously.Twenty-four cas-es of diabetic models, developed in mice with streptozotocin, were divided into 3 groups randomly.In experimental group, donor rat hepatocytes were infused first, followed by transplantation of cultured islets from same donor via portal vein on the 6th day.In Control group 1 no intervention was performed, and in control group 2, simple infusion of islets was done.The results showed group.Significant difference was noted between the two groups(P＜0.05).The decreased blood glucose level were maintained about a week in experimental group.This result reveals that hepa-tocytes can induce immunotolerance and prolong the survival of xenografts to some extent.

Objective To study the characteristics of granulomatosis with polyangiitis (GPA) accompanied by pneumothorax.Methods We described a case of GPA accompanied by hydropneumothorax who was successfully treated.Relevant literature was also reviewed.Results A total of 25 cases were identified,consisting of 18 males and 7 females [the average age was (44±16)(16-70) years old].The time from disease onset to pneumothorax was 26±51 (0.83-216) weeks.Pneumothorax,hydropneumothorax,pyopneumothorax and hemopneumothorax occurred in 11,5,8 and 1 respectively.Nodules or excavated nodules on chest radiography or CT were seen in 22 cases.Erythrocyte sedimentation rate (ESR) and C reactive protein (CRP) were elevated in all cases.Sixteen cases received glucocorticoid and immunosuppressive agents treatment.Sixteen cases received drainage and 7 received open operation.Pseudomonas aeruginosa was the most commonmicrobiology findings.Granulomatosis with active vasculitis,bronchopleural fistula,pleural bleb with intensefibrosis,rupture of subpleural nodule were seen on lung biopsy or autopsy.Nine cases died of infections,respiratory failure,sepsis and respiratory arrest.Conclusion Pneumothorax in GPA can be caused by multiple factors such as rupture of subpleural nodule and with high mortality.Patients always died of infections and respiratory failure.Regular treatment of the underlying disease,apply sensitive antibiotics for infection and reasonable surgical intervention should be considered.

Objective To explore the feasibility and secure cold storage time of human arteries during sequentially cold-cryopreservation by observing the cellular metabolic activity and structure after cold storage and cryopreservation. Methods Human iliac and splenic arteries were stored for 72 h, 1 week, 2 weeks, 3 weeks and 4 weeks in UW solution at 4 ℃. After the cold storage procedure, half of the vascular allografts were examined by NBT dye method, electron and light microscope. The other vascular allografts continued to be stored by - 80 ℃ cryopreservation procedure for 4 weeks, and then the vascular allografts were examined by NBT dye method, electron and light microscope. Results There was no statistically significant difference in NBT dyeing time between the groups stored in UW solution within 2 weeks and fresh group at 4 ℃ (P ＞ 0. 05). After - 80 ℃ cryopreservation, there was also no statistically significant difference in NBT dyeing time between the groups stored by UW solution within 1 week and fresh group at 4 ℃ (P＞0. 05). Along with the extension of cold storage time, the destruction of ultrastructure was aggravated. When vascular allograft was stored over 2 weeks at 4 ℃, the destruction was more obvious. As the cold storage time prolonged, the ultrastructural destruction of vascular allografts was aggravated, especially those stored over 1 week. Conclusion The optimal time limit for arteries stored at 4 ℃ in UW solution was 2 weeks. Cryopreservation at - 80 ℃ kept the arteries satisfactory metabolic activity and organizational structure. The arteries stored within 1 week at 4 ℃ in UW solution, which restored at - 80 ℃ , could maintain satisfactory metabolic activity and organizational structure.

Objective To investigate the effect of NR2B antisense oligonucleotide on naloxone-induced withdrawal responses in morphine-dependent rats. Methods Famale SD rats weighing 230-270 g were anesthetized with intraperitoneal pentobarhital 60 mg/kg. Intrathecal (IT)catheter was placed at L3,4 interspace.Thirty-two rats in which FT catheter was successfully placed were randomly divided into 4 groups ( n = 8 each) : group C control; group MD morphine dependence; group AO NR2B antisense oligonucleotide (aNR2B) and group SO NR2B sense oligonucleotide (sNR2B) . In group MD, AO, SO chronic morphine dependence was induced by increasing doses of subcutaneous morphine for 6 days. The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg twice every other day and reached 50 mg/kg on the 6th day. In group AO and SO IT aNR2B or sNR2B 15 nmol was administered simultaneously with subcutaneous morphine. Morphine withdrawal responses was induced by IT naloxone 4 mg/kg and scored based on the responses (0 = normal; higher scores signify severer responses) . The weight loss was calculated.The expression of NR1, NR2A and NR2B mRNA in hippocampus was determined by RT-PCR. Results The morphine withdrawal syndrome and weight loss were significantly incresed in group MD, AO and SO, while NR2B mRNA expression in hippocampus was up-regulated in group MD and SO compared with group C. The morphine withdrawal syndrome and weight loss were significantly decreased, NR2A mRNA expression in hippocampus was up-regulated and NR2B mRNA expression was down-regulated in group AO compared with group MD. There was no significant difference in NR1 mRNA expression between the 4 groups . Conclusion NR2B antisense oligonucleotide can suppress morphine withdrawal responses through the regulation of NMDA receptor level and construction in hippocampus.

Objective To investigate the effects of surgical trauma on synaptic structure in hippocampal CA3 area in aged rats.Methods Fifty-six healthy SD rats aged 18 months were randomly divided into 3 groups: control group (group C, n = 8) , anesthesia group (group A, n = 24) , and operation group (group O, n - 24) . Anesthesia was performed with intraperitoneal ketamine 40 mg/kg but no operation was carried out in group A. Anesthesia was also performed with intraperitoneal ketamine 40 mg/kg, and splenectomy was performed after loss of righting reflex in group O. Eight animals from group A and O selected on 1, 3, and 7 d after anesthesia or operation respectively underwent Morris water maze test for assessment of the cognitive function. The animals were . then decapitated. Hippocampal CA3 area was isolated for examination with electron microscope and the synaptic structure in the polymorphic layer of hippocampal CA3 area was measured. Results Compared to group C and A, the times of passing through the original platform and number of synapses were significantly reduced, the width of synaptic cleft was significantly increased, the thickness of the postsynaptic density was significantly decreased, the length of the active zones was significantly shortened, and the curvature of the synaptic interface and percentage of perforated synapses were significantly decreased at T_(1,2), ( P < 0.05 or 0.01) , but no significant change was found in the indices mentioned above at T_3 in group O(P > 0.05). Compared to group C, the latency and swimming distance were significantly prolonged at T_1 in group A and at T_(1,2) in group O ( P < 0.01) . Compared to group A, the latency at T_(1,2) and the swimming distance at T_2 were significantly prolonged in group O ( P < 0.05) . Conclusion Surgical trauma can induce early postoperative cognitive impairment through changing synaptic structure in hippocampal CA3 area in aged rats.

Objective To evaluate the diagnosis and treatment of vipoma based on our experience on 4 cases.Method Clinical manifestations,laboratory examination,imaging features,surgical findings,and pathology of 4 patients with vipoma admitted in our hospital from 1991 were discussed.Results Watery diarrhea and hypokalemia were the main clinical manifestations.Hepatic metastasis OCCurred in two patients.Tumor located in the head of the pancreas in one case.Two tumors were shown in the pancreatic body and one tumor was in the pancreatic tail.Resection of tumor and hepatic metastatic lesions with repeat resection of metastases Was performed in 1 patient.Resection of the pancreatic body and tail was done in one patient.Pancreatoduodenectomy Was performed in one patient.Laparotomy only was done in one patient because of invasion of the superior mesenteric vein and duodenum.Conclusion Typical symptoms play an important role in the diagnosis of vipoma.Hepatic metastasis is common.Surgery is the most effective means for treatment.

Objective To investigate the inhibitory effects of Argatroban on the instant bloodmediated inflammatory reaction(IBMIR)after islet transplantation.Methods Rat islets were isolated and purified rat islets,and were divided into blank control group,control group and experimental group.In the control group,the blood and the islets were directly mixed,and in the experimental group the Argatroban was added to the mixture based on the control group.while the blank control group was added with blood alone without the islets.Each group was reacted at 37℃for 60min,and then the content was filtered through trap valve of 70 μm.The residual thrombus and tissues were filtered by the trap valve in both the experimental and control groups,detected by the thinprep cytologic test(TCT),and the filtrate received blood routine test,and the function of islet was determined using insulin releasing test.Results The number of blood platelets,white blood cells,mononuclear cells,and lymphocytes percentage in the experimental group were significantly higher than in the control group after 60 min(P＜0.05).Under the environment of the high and low concentrations of glucose,the insulin release in experimental group was significantly increased as compared with the control group,and the insulin release index of former was 2.25±0.18,significantly higher than that of the latter 3.36±0.18(P＜0.05).The residual thrombus and tissues had few islet cells in the control group,the structure was damaged seriously,the capsule was not intact,and there were a large mumber of micro-thrombosis around the islets formed by red blood cells.But there were a large number of islet cells in the experimental group.the structure was intact in a mass,and no obvious micro-thrombosis around the islets was found.Conclusion Argatroban can effectively inhibit IBMIR in vitro,and alleviate the damage to the islet cells.

Objective To discuss the efficacy of GP combined with cervus blood granule in treating advanced non -small cell lung cancer (NSCLC ) after chemotherapy.Methods Eighty patients with advanced NSCLC were randomly divided into two groups by digital table method.In the combined group,forty patients were trea-ted by GP combined with cervus blood granule.In the control group,forty patients were treated by GP alone.All the patients were tested routine blood in the seventh and tenth day.The value of platelet was recorded,the incidence of thrombocytopenia between two groups was compared.Results At the seventh day of chemotherapy,in the control group,thrombocytopenia Ⅰ found in 5 cases,2 cases of thrombocytopeniaⅡ,1 case of thrombocytopeniaⅢ,thrombo-cytopenia Ⅳ in 1 case.In the combined group,thrombocytopenia Ⅰ occurred in 3 cases,1 case of thrombocytopeniaⅡ,no grade 3 or 4 thrombocytopenia,there was no statistically significant difference between the two groups(Z=-1.259,P=0.208).At the tenth day of chemotherapy,in the control group,thrombocytopenia Ⅰ found in 6 cases, 3 cases of thrombocytopenia Ⅱ,1 case of thrombocytopenia Ⅲ,thrombocytopenia Ⅳ in 1 case.In the combined group,4 cases occurred thrombocytopenia Ⅰ,thrombocytopenia Ⅱ in 1 case,no grade 3 or 4 thrombocytopenia.The difference between the two groups was statistically significant(Z=-1.966,P=0.049).Conclusion Cervus blood granule combined with GP can effectively reduce the classification of thrombocytopenia caused by chemotherapy.To some extent,it can also prevent the happening of the thrombocytopenia.

Objective Pancreas perfusion is an essential step in human islet isolation.To develop the new methods for introductal canulation,collagenase infusion and to observe their effects on islets isolation.Methods A total of 17 pancreases were digested from March 2005 to April 2010.The pancreases were distended by three methods:the standard method (n =3),the one-cannula method (n =11) and three-cannula method (n =3).In the standard group,the pancreases were completely cut into half at the mid-body.Two catheters were inserted into the main duct:one directed toward the tail and the other to the head.In the one-cannula method group,a long tube was inserted into the duct at the head,advancing to the tail In the three-cannula method group,pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct:one at the head (the first catheter) and two at the mid-body,one toward the tail (the second catheter) and the other toward the head (the third catheter).The pancreases were digested by improved Ricordi technique.Ficoll continuous density/grads centrifuge method was performed to purify the islets.DTZ staining was adopted to identify islets and count islet equivalent (IEQ). AO/EB fluorescence examination was used to count active islet percentage.Static glucose stimulating test (SGS) in vitro was designed to estimate islet function and calculate SI.Results The distension volume of the threecannula method group was 1.24 rnl/g pancreas,and higher than the other groups (for the standard group:0.71 ml/g pancreas; for one-cannula method group:0.96 ml/g pancreas,P＜0.05).The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g,which was significantly more than that in the standard group (1914 IEQ/g pancreas,P＜0.05).The purity and viability of the islets were 74 ％/79.3 ％,75.6 ％/79.4 ％ and 78.3 ％/84.0 ％ respectively in the three groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).ConclusionThe three-cannula method improved collagenase infusion and the islet yields.