Objective To investigate effects of microRNA-506 (miR-506) on malignant phenotypes of hepatocellular carcinoma (HCC) cells, including cellular viability, proliferation and invasion. Methods HCC cell lines HepG2 and QGY-7703 were served as model. Five experimental groups were established in this study, including cell control, pcDNA 3 blank vector control, miR-506 over-expression, pSIH1 blank vector control and miR-506 suppression groups. Real-time reverse transcription PCR assay was performed to measure miR-506 level. CCK-8, colony formation and Transwell assays were performed to detect viability, colony formation activity and invasion activity of HCC cell lines, respectively. Effects of miR-506 on these indexes were evaluated. Results In HepG2 and QGY-7703 cell lines, miR-506 level increased in the miR-506 over-expression group (P<0.01), and its level decreased in the miR-506 suppression group (P<0.05) compared with the related blank vector control groups. In the miR-506 over-expression group, cellular viability was significantly reduced (P<0.01), cell colony number decreased, and number of cell penetrating Transwell microporous membrane was also decreased (P<0.01). In the miR-506 suppression group, cellular viability significantly increased (P<0.01), and both colony number and penetrating cell number increased (P<0.05). Also, there were no effects on the above indexes in pcDNA3 and pSIH1 blank vector control groups compared with those of cell control group (P>0.05). Conclusion miR-506 plays a tumor suppressor role in HCC cells by inhibiting cell viability, colony formation and invasion.

Objective To discuss the causes of acute hematogenesis disorder after liver trans-plantation. Methods A retrospective analysis was performed to identify 6 patients with high fever,skin rash and acute haematogenesis disorder during 2005-2006 in our center. Results The 6 patients had viral infection, immunity damage, intake of marrow toxicity medicine and multiple organ dysfunc-tion syndrome after the operation. These might cause the disorder. Conclusion Infection, virus,drugs, graft versus host disease and immunity damage are the main reasons for acute hematogenesis disorder after liver transplantation.

Objective To investigate the risk factor for new-onset diabetes after transplantation (NODAT) and the relationship between NODAT and arterial stiffness. Methods Oral glucose tolerance test (OGTT) was performed on 195 patients with renal transplantation. The degree of arterial stiffness, which was determined by brachial ankle pulse wave velocity (baPWV), anklebrachial blood pressure index (ABPI) and intima-media thickness (IMT) of the carotid artery, was evaluated. Results Twenty-nine patients diagnosed as NODAT had significantly higher fasting plasma glucose before transplantation, blood pressure and incidence of hepatitis C virus (HCV) infection than in patients without NODAT. Multivariate regression analysis revealed that the risk factor of NODAT was fasting plasma glucose pre-transplantation, HCV infection and systolic blood pressure.The independent determinant of the advanced arterial stiffness on NODAT was the statement of hypertension and age. Conclusions High fasting plasma glucose prior to transplantation, HCV infection and high blood pressure are risk factors for NODAT in patients after renal transplantation.Strict control of blood pressure is the key way to prevent the NODAT and atherosclerosis.

Objective To investigate the repair and protective effect of extracorporeal membrane oxygenation (ECMO) on liver and bile duct after cardiac death in pig.Methods Eight pigs were purchased and cardiac arrest was induced by the administration of 1 g KCL intravenously,followed by 30 min cardiopulmonary resuscitation according to standard guideline.Cannulas were placed through inferior vena cava and abdominal aorta,and then connected to ECMO extracorporeal circulation pipes.ECMO was performed for 4 h.Circulation flow rate of hepatic artery and bile production were monitored and recorded.Lactate dehydrogenase (LDH),γ-glutamyl transferase (γ-GT) and direct bilirubin (DBIL) in bile were detected.Transaminase,tumor necrosis factor-α (TNF-α),interleukin-1β (IL-13),hyaluronic acid (HA),endothelin-1 (ET-1) and nitric oxide (NO) in serum were detected.Pathological change was observed by HE staining under optical microscope and cell apoptosis was detected by TUNEL.Results There was no bile production after cardiac death,which increased to 80％ of the baseline after 4h of ECMO.In addition,γ-GT,LDH and DBIL content in bile was (23.3 ± 11.8) IU/L,(15.9 ± 3.3) IU/L and (72.3 ± 21.4) mmol/ L,and IL-1,TNF-α and HA content in serum was (117.6 ± 39.0) ng/L,(120.4 ± 16.5) ng/L and (63.7 ± 4.4) ng/L,respectively,and no statistically significant differences were observed when compared with the baseline (all P ＞ 0.05).ET-1 content was (4.9 ± 1.3) ng/L and NO content was (135.3 ± 16.7)mmol/L in serum,which was statistically increased (both P ＜ 0.05).Pathological changes of liver and bile duct were significantly alleviated.Conclusion ECMO could exert protective effect on liver and bile duct after cardiac death.

Objective To investigate the changes in cyclophilin A (CyPA) signal pathway in rat myocardial tissue after cardiopulmonary resuscitation (CPR).Methods Sixty-eight healthy male Sprague-Dawley (SD) rats were randomly divided into sham operation group (n =8),instantancous CPR after cardiac arrest (CA),immediate CPR after CA (CRP),and 15,30,60 and 120 minutes after CPR groups,respectively,with 10 rats in each group.Asphyxia was simulated by occlusion of the tracheal tube at the end of exhalation.Mechanical ventilation,compression and epinephrine injection were given for restoration of spontaneous circulation (ROSC) in order to reproduce cardiopulmonary resuscitation after cardiac arrest (CA-CPR) models in rats.Hemodynamic changes were recorded at different time points.The blood was collected from abdominal aorta,and myocardial tissue was also harvested.The serum CyPA and CD147 levels were determined by enzyme-linked immunosorbent assay (ELISA).The protein positive cells and mRNA expression of CyPA and CD147 in myocardial tissue of the rats were determined by immunohistochemical and real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR).The protein expressions of matrix metalloproteinases (MMP-2,MMP-9) and myeloperoxidase (MPO) in rat myocardial tissue were determined by Western Blot.The neutrophil infiltration in rat myocardial tissue was assessed by hematoxylin and eosin (HE) staining.Results The heart rate of rat was lowered to 0 with arterial pressure lowered immediately after CA.Arterial pressure was elevated to normal range immediately after CPR.Heart rate was restored at about 30 minutes later,and the dose of epinephrine was 50-60 μg,and ROSC time was 1-4 minutes.Compared with those of the sham group,serum CyPA and CD147 levels were gradually increased along with elongation of ROCS time within 120 minutes,and peaked at 120 minutes,CyPA was increased from (786.11 ± 3.93) μg/L to (2 001.80 ± 10.61) μg/L,and CD147 was increased from (2.94±0.02) μg/L to (5.99±0.023) μg/L (P ＜ 0.05 or P ＜ 0.01).CyPA and CD147 mRNA expressions (A value) in rat myocardial tissue were gradually increased,and peaked at 120 minutes.Relative expression of CyPA at 120 minutes was 2.42 ± 0.05 when it was 1 in the control,and relative expression of CD147 at 120 minutes was 1.88 ±0.10 (both P ＜ 0.01).The immunohistochemical results under light microscope showed that the brown positive cells were gradually increased,which indicated that the expressions of CyPA and CD147 were increased.Expressions of MMP-2,MMP-9 and MPO (gray value) in myocardial tissue were also gradually increased,peaking at 120 minutes,and MMP-2 was increased from 0.396 ± 0.021 to 0.879 ± 0.020,MMP-9 was increased from 0.372 ± 0.009 to 0.819±0.012,and MPO was increased from 0.176±0.005 to 0.829±0.018 (P ＜ 0.05 or P ＜ 0.01).No obvious neutrophil infiltration in myocardial tissue was found with HE staining.Conclusion Expressions of CyPA and CD 147 were up-regulated in serum and myocardial tissue after CPR in rats,which may be the markers of inflammatory reaction.

Objective:To follow up the hepatitis B recurrence post liver transplantation using the combination of lamivu dine and high-dose intravenous/intramuscular hepatitis B immunoglobulin.Methods:Fifty-two consecutive patients who underwent liver transplantation were included in this analysis.All patients were administered lamivudine combined with hepatitis B immunoglobulin intravenous(V group)or intramuscular(M group)injection prophylaxis.Dosages of hepatitis B immunoglobulin were 4 000 U during anhepatic phase,2 000 U/d during the first six days after operation,2 000 U/week at 2-4 weeks after operation and 2 000 U/month at 2-6 months after operation.The dosages of hepatitis B immunoglobulin were decided by blood concentrations at 6 months after transplantation in both groups(HBsAb titer≥200 U/L during the 6-12 months after operation;100-200 U/L one year after operation).The negative-conversion rates of serum HBsAg,serum HBsAb levels and hepatitis B recurrence rate at the 6th month,1 year after transplantation and at the end of this study were investigated.Results:The daily average negative-conversion rate of serum HBsAg was significantly faster in V group than that of M group within the first 4 postoperative days(P ＜ 0.05).The 100% of serum HBsAg negative-conversion rate was obtained in two groups after one week of operation.No recurrence was found in two groups.And the ease of death was independent on HBV recurrent during the follow-up.Conclusion:Lamivudine combined with hepatitis B immunoglobulin intravenous/intramuscular injection prophylaxis was effective protocol for the hepatitis B recurrence after the liver transplantation.

BACKGROUND: The formation mechanism of biliary cast syndrome following liver transplantation has not been thoroughly illuminated, and it is unclear that whether some proteins correlated to the formation mechanism of biliary cast or prewarning to the formation of biliary cast.OBJECTIVE: To investigate the holoprotein expression in biliary cast syndrome patients following liver transplantation. METHODS: Four patients underwent liver transplantation at Liver Transplantation Institute, General Hospital of Chinese People's Armed Police Force. Three months later, 10 g biliary cast was harvested. Four kinds of biliary cast specimens at different colors and textures were preserved at deep hypothermia, followed by protein abstraction and restriction enzyme digestion, the total protein abstraction solution of biliary cast were analyzed by high definition mass spectrometry and query on MASCOT database. All protein name of biliary cast were list, the conjunct protein was found by comparing 4 specimens. RESULTS AND CONCLUSION: There were totally 208 proteins in 4 biliary cast specimens, 82, 44, 56 and 65, respectively. By comparison, 5 proteins were found to overlay in 2 biliary cast specimens, 7 proteins in 3 specimens and 13 proteins in 4 specimens. Among the latter 13 proteins, 5 unnamed-proteins, as well as 8 named-proteins (termed alpha-fibrinogen precursor, beta-fibrinogen precursor, fibrinogen gamma chain, proapolipoprotein, Chain A of Human Cathepsin G, S100 calcium-binding protein A9, lactoferrin) were included. The proteins exists in biliary cast, the common proteins of 4 biliary cast specimens imply a correlation between the formation of biliary cast and the exudative inflammation following the damage of biliary tract epithelium; Some proteins might be considered as a marker of prewarning the presence of biliary cast syndrome, judging the inflammation severity following the damage of biliary tract epithelium and the prognosis of biliary cast syndrome.

Objective To investigate the optimal time of extracorporeal membrane oxygenation (ECMO) on repairing Maastricht Ⅱ pig liver with warm ischemia injury for 30 min.Method Thirty-six miniature pigs were randomized to ECMO 2-h group,ECMO 4-h group and ECMO 6-h group,12 pigs per group,6 donors and 6 recipients.Cardiac arrest was induced by administration of 1 g KCl intravenously,followed by cardiopulmonary resuscitation (CPR) for 30 min according to the standard guideline.Cannulas was placed through inferior vena cava and abdominal aorta,then connected to ECMO extracorporeal circulation pipes.Transaminase,circulation flow rate of portal vein and hepatic artery and arterial blood gas were monitored and recorded.The hepatic tissues were cut into sections for pathological observation by HE stain under a light microscope.Apoptosis was detected by TUNEL and apoptotic index was calculated.The liver was stored in cold UW for 2 h after the ECMO circulation,then orthotopic liver transplantation without veno-venous bypass was performed.The levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured in the peripheral blood for 5 days after the operation.Result With the increase of the running time of ECMO,transaminase and lactate levels were decreased continuously.Circulation flow rate of portal vein and hepatic artery in ECMO 6-h group was significantly lower than that in the other two groups (P＜0.05).Pathological change in ECMO 4-h group was milder than the rest two groups.Apoptosis index in ECMO 2-h,4-h and 6-h groups was (40.20 ± 7.22)％,(18.60 ± 4.04)％ and (29.25 ± 5.98) ％,respectively.The 5-day suvival rate in ECMO 2-h,4-h and 6-h groups was 83％,100％ and 83％,respectively.The transaminase level in ECMO 4-h group at 5th day after the operation was lower than in ECMO 2-h group and 6-h group (P＜0.05).Conclusion The optimal time of ECMO on repairing Maastricht Ⅱ liver was 4 h.The effect of restoration is not ideal when circulation time is not enough.Liver function and liver cell viability decline beyond 4 h.

Objective To study the repair mechanism of extracorporeal membrane oxygenation on liver after cardiac death.Methods Twelve pigs were equally randomized to ECMO group and control group.Cardiac arrest was induced by administration of 1 g KCL intravenously,followed by 30 min cardiopulmonary resuscitation according to the standard guideline Cannulas were placed through inferior vena cava and abdominal aorta,then connected to ECMO extracorporeal circulation pipes in ECMO group for 4 h.The livers were stored in cold UW for 4 h in control group.ATP,superoxide dismutase (SOD),glutathionein (GSH),malondialdehyde (MDA),heat shock protein 70 (HSP70) and intercellular cell adhesion molecule-1 (ICAM-1) were detected in liver tissue.Pathological change was observed by optical microscope and electron microscopy.Results Tissue ATP decreased to less than 40％ of baseline after 30 min of warm ischemia,then restored to 70％ after 2 h of ECMO and returned to baseline after 4 h,while ATP of control group continued a further decline As compared with control group,SOD,GSH and HSP70 increased significantly in ECMO group (P＜0.05),while MDA and ICAM-1 decreased significantly (P＜0.05).Pathological changes of liver tissue observed by optical microscope and electron microscopy in ECMO group were significantly were significantly alleviated as compared with those in control group.Conclusion ECMO can supply oxygen and nutrients to liver after warm ischemia and increase energy reserve.By upregulating GSH,SOD and HSP-70 and other protective proteins,ECMO alleviates oxidative stress and liver damage ECMO also improves microcirculation and reduces neutrophil infiltration by protecting sinusoidal endothelial cells.

Objective To establish a stable and reliable model of brain death in swine, and to provide a more stable model for investigating pathomorphology in brain tissue and for studying transplantation immunology during brain death. Methods Base on the classic methodology of increasing epidural intracranial pressure, Codman intracranial pressure moni-toring probes were implanted in landrace pigs invasively. According to the relationship between ICP and MAP, brain death models were established by increasing intracranial pressure slowly and intermittently, with real-time monitoring of the intra-cranial pressure. Results Among twelve experimental landrace pigs, one died from anesthetic accident, while the rest elev-en were successfully established as brain death models. With effective respiratory and circulatory support, those brain death models can be maintained for (31.3 ± 4.7) h. Brain death model establishement is a stable and reliable process demonstrated by transcranial Doppler, EEG, ECG, mean arterial blood pressure and other monitoring methods. After brain death is con-firmed, animal models can be maintained for a long time. Conclusion Our methodology of inducing brain death model un-der ICP monitoring is stable and easy to be standardized. It can also provide a more stable model for studying brain tissue pathomorphology and transplantation immunology.