Abstract:

Objective: To evaluate the effectiveness of interventional therapy (IT) and timing of retransplantation for treatment of hepatic artery stenosis (HAS) after orthotopic liver transplantation (OLT).
Methods: The clinical data of 20 patients diagnosed as hepatic artery stenosis (HAS) were retrospectively analyzed.Among them, 6 cases underwent retransplantation. HAS was confirmly by digital subtraction angiography (DSA). All patients were treated with percutaneous interventional management including percutaneous transluminal angioplasty (PTA) and stent-graft placement.
Results: After IT in the 8 patients with early HAS (within 1 month of transplantation),  liver function improved in 6 cases,one underwent retransplantation due to deterioration of liver function, and one died of acute liver failure during awaiting a proper liver for retransplantation.After IT in the 12 patients with late HAS (after 1 month of liver transplantation)，one died of severe sepsis 38 days after transplantation, five  patients underwent late retransplantation due to ischemic-type biliary strictures or recurrent attacks of cholangitis, and one of these patients died 11 days after retransplantation. The median follow-up of 17 patients was 13 months after liver transplantation. Two patients died of severe cholangitis and recurrence of hepatocellular carcinoma unrelated to HAS,and 3 patients developed recurrent hepatic arterial stenosis and were successfully treated with second interventional therapy. Eight patients (40%) developed ischemic-type biliary strictures and underwent interventional treatment. Graft function in 5 of the 8 patients improved.  The Kaplan-Meier curve of survival showed the 1，and 2 year cumulated survival rates of early HAS and late HAS were 87.5%，43.8% and 81.5%，54.3%，respectively. There was no significant difference in 1 and 2 year survival rates between early HAS and late HAS (log-rank test，P=0.976).
Conclusions: Interventional therapy is the treatment of choice for HAS after OLT when the HAS has not resulted in severe hepatic dysfunction, but the incidence of ischemic biliary tract stenosis is relatively high. Liver transplantatation is the only effective theropy when HAS induced irreversible hepatic dysfunction.

Abstract:

Objective: To stud the clinical application of improved bile duct reconstruction in liver transplantation.
Methods: We retrospectively analyzed the clinical data of 131 patients who received liver transplantation from September 2005 to September 2007. The patients were divided into two groups according to the method of bile duct reconstruction. Group A (n=76) received traditional bile duct reconstruction, group B (n=55) received improved bile duct reconstruction. All operations were done by one operation team. The 2 groups were similar in age, gender, liver transplantation indications and Child-Pugh score. We mainly analyzed the time of bile duct reconstruction and postoperative biliary complication associated with anastomsis.
Results: We spent 14±2 min to reconstruct the bile duct in the traditional method group, and (13±2) min in the new method group (P<0.05).In A group, 4 cases (5.3%) encountered the biliary complication associated with anastomsis, 2 cases suffered from biliary strictures were cured by ERCP(endoscopic retrograde cholangiopancreatography); two cases suffered from bile leaks were recovered by conservative therapy. In B group, 1 case (1.8%) suffered from biliary complication associated with anastomsis was cured by ERCP.
Conclusions: The new bile duct reconstruction is easier and has fewer complications. It may be an ideal method for bile duct reconstruction in liver transplantation. 

Abstract:

Objective: To investigate the risk factors and the clinical findings associated with fungal infection during early postoperative period after liver transplantation (LT).
Methods: The clinical data of 118 cases of liver transplantation performed during past four and a half years in Nanfang Hospital were studied retrospectively. Forty-four independent variable factors were analyzed by univariate analysis and logistic regression to screen out the risk factors for early fungal infcction after LT.
Results: Fungal infections occurred in 26 cases（22.0%）, and the mean time was 13.6 d. A total of 49 fungi strains were isolated, of which mainly was Candida albicans(57.1%).The most common infection site was the respiratory tract (75.5%).Logistic regression analysis showed that pre-operative liver failure, duration of mechanical ventilation over 48 hours, pleural effusion, observation in ICU for more than 5 days, and the use of more than 3 kinds of antibiotics and for longer than 2 weeks were risk factors. The infection rate was lower in patients with prophylactic administration of antifungal ugents than patients without prophylactic administration.（P＜0.05）.
Conclusions: Because multiple risk factors can lead to fungal infections following liver transplantation, it is important to make effective control of these factors and thus reduce the incidence of fungal infection. The use of antifungal agents for prophylaxis in high-risk LT patients is associated with a low incidence of serious fungal infection.

Abstract:

Objective: To investigate the expression and role of early growth response (EGR-1) in early injury of small-for-size graft in rat liver transplantation.
Methods: Male Sprague-Dawley rats were randomly divided into three groups: Sham  operation group （SO group）; small-for-size graft group(30%graft group); whole graft group(100%graft group). The models of rat small-for-size liver transplantation were set up by two-cuff technique. Changes of AST in serum, pathology of liver, expression of EGR-1 and endothelin (ET)-1 in liver of three groups within 24 hours after reperfusion were compared.
Results: AST of SO group was significantly lower than that of the other two groups（P＜0.01），while AST of 100% graft group was lower than that of 30% graft group，especially at 6h and 24h after reperfusion （P＜0.01）. Apparent dilation of the sinusoid with disintegration and detachment of sinusoidal lining cells were observed in 30%graft group at 24h after reperfusion. There was no expression of EGR-1 and ET-1 in SO group, whereas expression of these two factors in 30%graft group  were significantly higher than that in 100% graft group within 24 hours after reperfusion.
Conclusions: Small-for-size graft injury was related to early over-expression of EGR-1 associated with upregulation of ET-1.

Abstract:

Objective: To investigate the reparative effect of HNF4α embryonic hepatic stem cells on acute hepatic injury. 
Methods: Suspension of ED 14 Fischer (F) 344 rat embryonic hepatic stem cells was prepared by collagenase digestion and mechanical disaggregation and maintained by long-term culture in vitro. Healthy SD female rats were randomly divided into transplantation group (TP group, n=20) and control group (n=20). The experimental acute hepatic injury was induced by CCl4-lavage. Suspension of embryonic hepatic stem cells was transplanted into injured livers of rats in transplantation group via portal vein while saline of equal volume was used in control group. The levels of ALT and AST in the serum and histopathological injury of hepatic tissues were analyzed 6 h before, and 1w, 3w and 5w after injection. Evaluation of HNF4α protein expression in rat livers by immunohistochemistry and Western blot was performed before and after treatment.
Results: The liver function of control group was slow recovery, as evidenced by significantly increased ALT and AST in serum（P＜0.05）. By contrast, the injured hepatic tissues were markedly repaired in TP group, and liver function gradually returned to normal level. The survival rate of TP group at 5 weeks was significantly higher than that of control group (P=0.002). After liver injury, expression of HNF4α in the injured liver tissue notably increased. And then, in accordance with the gradual restoration of liver tissue, HNF4α expression was slowly decreased.
Conclusions: HNF4α plays a protective effect in the early stage of hepatic injury after liver transplantation, and it could probably act through promoting the restoration of acute hepatic injury by facilitating the differentiation, proliferation, and migration to damaged hepatic lobule of embryonic hepatic stem cell.

Abstract:

Objective: To investigate the protective effect of the first window (FW)of liver ischemic preconditioning(IPC), the second window(SW) of remote (leg) ischemic preconditioning(RPC) and conbined applications of liver and lges IPC to against liver ischemia/reperfusion(I/R) injury in the rat, and to investigate the mechanism of the protection.
Methods: Rats were randomly divided into five groups (n=8 each):(1) Sham group (S group), rats without IPC, (2) Rats with 5 min IPC (IPC group); (3) Rat wiht both liver and lower limbs IPC and repeated three times (RPC group); (4) IPC 24 h after RPC group; (5) IR without IP (I/R group)； except S group, the rats were subjected to 60 min sustained liver ischemia followed by 180 min reperfusion.All ischemia rats were only subjected to 70% liver ischemia. Finally，blood and liver samples were obtained to determine the activity of ALT and AST, the expressions of TNF-α and HSP70  protein,and liver wet/dry weight (W/D) and pathology.
Results: All IPC group and RPC group and IPC+RPC group had obviously lower levels of ALT, AST,  W/D, TNF-α than that of the I/R group (P＜0.01), but had obviously higher expressions of HSP70 protein than the I/R group (P＜0.01), howerer,all the indices between IPC group and RPC group and IPC+RPC group had no statistical  difference(P＞0.05).
Conclusions: The FW of the IPC, the SW of the RPC and combined applications can lessen hepatic I/R injury. There is no significant difference in the protective intensity of the 3 motheds. The protective effects possibly are due to suppression of TNF-α production, induction of protein HSP70 expression and improvement of liver microcirculation.

Abstract:

Objective:  To investigate the protective effect of edaravone on ischemia reperfusion injury (IRI) in rat liver.
Methods: Sixty rats were randomly divided into experimental group and controll groups (30 in each) after establishing animal liver IRI model with partial reperfusion injury under normal temperature.Just after initiation of reperfusion and 1h later,edaravone was administered in the experiment group,and the same volume of normal saline was administered in the control group.The lipid peroxidation (LPO) hepatic enzymes and the level of TNF-α mRNA and E-selectin mRNA in plasma were measured at 0, 2, 4 h after initiation of reperfusion.We also serially quantified hepatic expression of mRNAs for TNF-α and E-selectin with RT-PCR.
Results: In the experiment group,hepatic LPO and hepatic enzyme were significantly less than that in the saline group(P＜0.05) at 2 h after initiation of reperfusion.Hepatic expression of TNF-α and E-selectin mRNA was significantly lower in the experiment group than the saline group(P＜0.05) at 2h after initiation of reperfusion.Histologically, E-selectin expression was less evident in hepatic sections in the experiment group than controll group at 2h after initiation of reperfusion.
Conclusions: Edaravone can reduce hepatic ischemia reperfusion injury,and significangly inhibit subsequent inflammation by reducing expression of inflammatory cytokines and adhesion molecules.

Abstract:

Objective: To investigate the protective effect of curcumin in hepatic cold ischemia-reperfusion injury in rats and its possible mechanisms.
Methods: Forty Wistar rats were randomly divided into two groups: the experimental group (n=21),rats were administered curcumin by injection through tail vein  at the dose of 60mg/kg which was dissolved in one milliliter DMSO two hours before cold ischemia period,and the remaining rats served as control group.The  hepatic cold ischemia was 30 minutes. All the rats were sacrificed six hours after reperfusion. Blood and liver samples were collected for determination of serum levels of ALT, AST and LDH and the tissue homogenate levels of SOD, MDA and MPO. Pathomorphological changes of liver tissue and the situation of apoptosis were observed under optical microscope. The content of TNF-α and MIP-2 in the liver homogenate were determined by enzyme linked immunosorbent assay(ELISA).
Results: Compared with control groups, the serum levels of ALT,AST and LDH in experimental groups were significantly decreased （P＜0.01）.The tissue homogenate levels of MDA,MPO, TNF-α and MIP-2 in experimental groups were significantly decreased.（P＜0.05）; the level of SOD was obviously elevated（P＜0.05）.Pathological studies indicated that curcumin attenuated liver tissue injury and inhibited apoptosis in the rats.
Conclusions: Curcumin can attenuate the cold ischemia-reperfusion injury of liver in rats, and its possible mechanisms may be related to increasing the activity of SOD,inhibiting lipid peroxidation and apoptosis,decreasing the expression of TNF-α and MIP-2, and suppressing the activation and infiltration of neutrophils.

Abstract:

Objective: To explore the possible mechanism of protective effects of curcumin against the ischemia reperfusion injury (IRI)  in liver autotransplantation  in rats.
Methods: A total of 54 male SD rats were randomly divided into three groups:curcumin preconditioning (CU) group,  0.5% sodium carboxymethycellulose preconditioning (CM) group, and the sham operation (SO) group.In CU and CM groups liver autotransplantation model were constructed.Then, 6 rats of each group were executed at 2, 6, 24 h after operation or liver reperfusion, respectively. Plasma samples were collected for ALT and AST test and liver tissues were harvested to detect the content of the MPO.
Results: The serum level of ALT and AST in CU group, and CM group was markedly elevated compared to SO group, but in CU group was  significantly lower than CM group at 2, 6, 24 h after reperfusion. Compared with SO group, the content of MPO was decreased significantly in CM group and CU group, and was significantly reduced  in CU group compared to CM group.
Conclusions: Curcumin preconditioning can attenuate liver IRI and the mechanism may be associated the decreasa of neutrophil infiltration.

Abstract:

Objective: To construct recombinant plasmid CD147-shRNA and detect its inhibitory effect on CD147 expression of hepatoma cell line SMMC- 7721 and HepG2.
Methods: Three characteristic target sequences were designed according to the sequence of CD147 in Genebank,and the synthesized polynucleotide was connected to pBS/U6.The recombinant plasmid was identified by restriction enzyme digestion and sequencing and transfected into hepatoma cells SMMC-7721 and HepG2. The expression of CD147 was detected by Western blot and immunofluorescence.
Results: CD147-shRNAs recombinant plasmid was completely and correctly constructed comfirmed by restriction enzyme digestion and sequencing. All the plasmids had gene silencing effect on CD147 expression of HepG2, but the inhibit effect of pBS/U6/CD147- shRNA3 was most significant.
Conclusions: We succesfully constructed pBS/U6/CD147-shRNA and screened out pBS/U6/CD147-shRNA3 which has the best gene silence effect.

Abstract:

Objective: To study the protective effect of NF-κB decoy oligodeoxynucleotides (ODNs) on endotoxin-induced liver injury in the rat and its mechanism.
Methods: Sixty Sprague-Dawley rats were randomly divided into control group,endotoxin (LPS) group and NF-κB decoy ODNs group. Liver and serum samples were collected 24 h after operation. The NF-κB binding activity was detected with electrophoretic mobility shift assay (EMSA), histopathologic change of liver was examined by light microscope, and cell apoptosis was detected by TUNEL. Liver enzyme (AST), TNF-α and IL-6 in serum were also detected.
Results: Compared to control group, in LPS group, NF-κB was dramatically increased induced apoptosis of large numbers of hepatocytes, and severe liver damage, the expression of AST, TNF-α and IL-6 were also significantly increased  (P＜0.01). However,compared with control group in NF-κB decoy ODNs group, the activation of  TNF-κB depressed, the hepatic histopathologic changes and apoptosis of hepatocytes were also relieved, and expression of  TNF-α and AST levels decreased (P<0.01). On the other hand, the expression of IL-6 was not significantly decreased by NF-κB decoy compared with LPS group (P＞0.05). 
Conclusions: NF-κB decoy strategy can efficiently inhibit the binding activity of NF-κB, and thus suppress the production of downstream cytokines, which may play a crucial role in protecting the liver from endotoxin-induced injury.

Abstract:

Objective: To observe the level changes of serum interleukin-18(IL-18) and tumor necrosis factor-α(TNF-α) in alcohol liver disease of rats.
Methods : The dose of 56% alcohol ［5～9 g/(kg·d)］  was administeredvia gastrolavage once daily for 12 weeks in ALD model rats. The control rats were grven the same volume of saline. The rats were killed at the end of 4，8, 12W. The pathological changes of liver were observed under light microscope after HE staining,and the levels of IL-18 and TNF-α in serum was determined with ELISA.
Results: The tissues of model rats showed various changes of chronic alcohol liver disease at the end of 4，8,12W,such as: fatty degeneration, inflammatory changes and fibrosis. The levels of ALT and AST in models were obviously higher than those of the controls(P＜0.05).The levels of IL-18 in models were obviously lower than the controls, and TNF-α was  higher,and the changes varied according to different degrees of liver damage.
Conclusions: The level changes of serum IL-18 and TNF-α in alcohol liver disease in rats are related to the degree of severity of the hepatitis, and indicate that  the development of the hepatitis relating to the changes of IL-18 and TNF-α.

Abstract:

Objective: To investigate the natural killer cell stimulatory receptor NKG2D and its soluble ligand MICA(sMICA) expression in peripheral blood in hepatocellular carcinoma (HCC).
Methods: Blood samples of 40 patients and 20 healthy persons were collected to detect NKG2D by flow cytometry analysis.Serum levels of sMICA were determined by ELISA.The association of sMICA levels wth clinical features were analyzed.
Results: The expression of NKG2D in HCC (71.97±4.96)% was significantly lower than that in healthy group(91.45±3.16)%. The serum levels of sMICA was found to be significantly elevated in HCC compared with heathy subjects (median, 191.3pg/mL vs. 74.9pg/mL，P＜0.05).The levels of sMICA correlated significantly with presence of tumor thrombi in portal vein and advanced clinical tumor-node-metastasis (P＜0.05）. Spearman rank correlation displayed that the expression of NKG2D was negatively correlated with the levels of sMICA（r=-0.591，P＜0.05）.
Conclusions:  The expression of NKG2D in HCC is significantly decreased. The levels of sMICA correlates significantly with progression of HCC. NKG2D-sMICA may play a critical role in tumor immunological escape in HCC,and may contribute to HCC progression.

Abstract:

Objective: To explore the  significance of matrilin-2 expression  in hepatic oval cells during the liver regeneration of rats.
Methods: The liver regeneration model of the rat was constructed by using partial hepatectomy(PH group) or  partial hepatectomy after 2- acetylaminofluorene gastrogavage(2AAF/PH group). The matrilin-2 expression of the 2 models was observed and compared with the normal rats(control group). We used immunohistochemistry and RT-PCR to dectect the expression of matrilin-2 in the models.
Results:  In control group and  partial hepatectomy group, immunohistochemistry showed that there was extremely limited amount of matrilin-2  which was  located around the vessels of bile ducts and within the hepatic sinusoids.However, matrilin-2 was observed within the hepatic sinusoids around the portal vein in the 2-AAF/PH group.  In addition, RT-PCR showed a high level  expression of matrilin-2 mRNA in hepatic oval cells of 2-AAF/PH model whereas matrilin-2 mRNA was not found in the PH group and control group.
Conclusions: Our results show that matrilin-2 is an important extracellular inatrix protein produced during liver regeneration,and is also closely relevant to the proliferation and differentiation  of hepatic oval cells.

Abstract:

Objective: To investigate the weight loss mechanism of sleeve gastrectomy.
Methods: SD rats were fed with high-fat diet to induce obesity, and then the obesity rats were randomly divided into experimental group (underwent SG) and control group (underwent sham surgery).Eight weeks after operation, the body weight, food intake and peripheral blood concentration of Ghrelin, GLP-1, PYY3-36 of rats were checked.
Results: Compared with controll group, in experimental SG model, food intake was decreased (P＜0.05); the postoperative weight continued to decline for three weeks and then slowly recovered, and body weight was still lower than the preoperative 8 weeks later,while Ghrelin decreased and GLP-1, PYY3-36 increased (P＜0.05).With time, Ghrelin gradually increased and GLP-1, PYY3-36  gradually declined. In the control group,compared with peroperation, the postoperative weight markedly increased except one week after operation, and  8 weeks later, weight was significantly higher than preoperation; but food intake, Ghrelin, GLP-1 and PYY3-36 had no significant change (P＞ 0.05).
Conclusions: The weight loss of sleeve gastrectomy in DIO rats’s decline is lasting and obvious. The postoperative effect of decreased Ghrelin and increased GLP-1, PYY3-36 is the main mechanism for weight loss.

Abstract:

Objective: To explore the clinical efficacy of hepatic segmentectomy under segmental staining and intraoperative chemoembolization for primary liver cancer(PLC).
Methods: Twenty cases of liver cancer underwent hepatic segmentectomy under segmental staining and intraoperative chemoembolization (observed group), the results were compared with 22 cases of PLC after treated by routine hepatectomy(control group).AFP,CT and MRI were regularly used after hepatectomy to evaluate the outcome.
Results: In observed group, the operative blood loss was(295±105)mL，blood transfusion was(280±85)mL,  liver function levels were in  the normal range accounted for 15%(3/20) one week  postoperatively, the incidence of postoperative complications was 40% (8/20),the postoperative 3-year survival rate was 60%,and the postoperative local recurrence rate was 35%; while in the control group, these parameters were (490±140)mL, (370±105)mL,  40.9%(9/22), 45.5％（10/22）,  40.91% and 68.18% respectively. In observed group, the operative blood loss, blood transfusion, cases with liver function levels in the normal range, the incidence of postoperative complications, postoperative 3-year survival rate, and postoperative local recurrence rate were significantly lower than those in the control group(P<0.05). The postoperative 3-year survival rate in observed group was significantly higher than that in the control group (P<0.05)，but the incidence of postoperative complications in the two groups was not statistically different (P>0.05).
Conclusions: The hepatic segmentectomy under segmental staining and intraoperative chemoembolization for PLC may reduce postoperative complications，lower postoperative relapse rate and improve survival rate.

Abstract:

Objective: To study the relationship between plasmic nitric oxide (NO), endothelin-1 (ET-1) levels and occurrence of hepatopulmonary syndrome (HPS) in liver cirrhosis with portal hypertension (LCPH) patients.
Methods: Fifty-two LCPH patients were divided into group Ⅰ（with HPS） and  groupⅡ (without HPS). Plasmic NO and  ET-1 levels were detected by nitric acid reductase and radioimmunoassay (RIA) respectively.
Results: Occurrence of HPS in all patients was 11.5%（6/52）, and was 30% (3/10) in Child-Pugh class C patients. In group Ⅰ, 50% (3/6) patients were in Child-Pugh class C. The comparison showed that plasmic NO levels were significantly higher in group Ⅰ than that in group Ⅱ［(110.40±28.02)μmol/L vs. (79.56±22.12)μmol/L, P＜0.01］, but there was no significant difference in ET-1 levels（P＞0.05）.
Conclusions: Elevated plasmic NO levels is an important characteristic of HPS, and, when combined with Child-Pugh classification, it can be significant for diagnosis of HPS in LCPH patients.

Abstract:

Objective: To evaluate the effects of ischemic preconditioning(IP) on liver function, complications and hospital stays after hepatectomy under hepatic vascular exclusion by a meta-analysis.
Methods: Randomized controlled trials (RCTs) were identified from PUBMED, EMBASE, the Cochrane Library, VIP, CNKI and Wanfang Data according to the inclusion and exclusion criteria. Literature screening, data extraction and quality assessment were made and the meta-analysis was processed by RevMan 4.2.2.
Results: Eight RCTs involving a total of 511 patients were included. The methodological quality was evaluated and all the trials were in graded B. The meta-analysis revealed that the postoperative ALT peak level (weighted mean difference=-176.37; 95%CI:-320.67~-30.06; P=0.02)and postoperative complications incidence (odd ratio=0.64; 95%CI: 0.41~0.98; P=0.04)were lower in IP group compared with control group, but there were no significant differences in blood loss, operating time, hepatic vascular exclusion time, postoperative AST and total bilirubin peak level, and hospital stays in both groups.
Conclusions: IP reduces the postoperative ALT peak level and complications incidence after hepatectomy under hepatic vascular exclusion, but there is no sufficient evidence to support that the IP can protect the liver from ischemia/reperfusion injury.