Strategies to rescue steatotic livers before transplantation in clinical and experimental studies

The shortage of donor livers has led to an increased use of organs from expanded criteria donors. Included are livers with steatosis, a metabolic abnormality that increases the likelihood of graft complications posttransplantation. After a brief introduction on the etiology, pathophysiology, categories and experimental models of hepatic steatosis, we herein review the methods to rescue steatotic donor livers before transplantation applied in clinical and experimental studies. The methods span the spectrum of encouraging donor weight loss, employing drug therapy, heat shock preconditioning, ischemia preconditioning and selective anesthesia on donors, and the treatment on isolated grafts during preservation. These methods work at different stages of transplantation process, although share similar molecular mechanisms including lipid metabolism stimulation through enzymes or nuclear receptor e.g. , peroxisomal proliferator-activated receptor, or anti-inflammation through suppressing cytokines e.g. , tumor necrosis factor-α, or antioxidant therapies to alleviate oxidative stress. This similarity of molecular mechanisms implies possible future attempts to reinforce each approach by repeating the same treatment approach at several stages of procurement and preservation, as well as utilizing these alternative approaches in tandem.

Warm HTK donor pretreatment reduces liver injury during static cold storage in experimental rat liver transplantation

BACKGROUND： Organ shortage has led to an increased number of transplantations from extended criteria donors. These organs are more vulnerable to ischemia-reperfusion injury. Thus, improvement of organ preservation is needed. HTK is a widely used preservation solution for static cold storage in liver transplantation. The present study was to investigate the beneficial effect of warm HTK donor pretreatment on liver preservation.

Application of two-phase helical CT in liver neoplasms

Objective: To assess the value of helical CT in the di- agnosis of liver diseases. Methods: 59 patients with different liver diseases were examined by two-phase or multi-phase dynamic helical CT. Results: Small hepatocellular carcinoma showed a higher density in the arterial phase, and a lower den- sity in the portal vein phase. Large hepatic carcino- ma showed a mixed pattern of higher-density in the arterial phase, and a lower density in the portal vein phase. Metastasis carcinoma showed an 'oxeye sign' in the portal vein phase. Hemangioma was not obvi- ously enhanced in the early arterial phase, marginal- ly enhanced in the arterial phase, and equally-densed in the balanced phase. Conclusion: Two-phase helical CT is of value in im- proving the detection rate of or determining the fea- tures of hepatic diseases by two-phase helical dyna- mic scan (2.0-3.0 ml/s speed, and delay time 25- 30 s and 70-85 s).

FEASIBILITY STUDY OF AN ULTRASOUND CONTRAST AGENT(LEVOVIST) IN COLOR DOPPLER IMAGING OF LIVER NEOPLASMS

ＩＮＴＲＯＤＵＣＴＩＯＮＣｏｌｏｒＤｏｐｐｌｅｒｉｍａｇｉｎｇ（ＣＤＩ）ｆｏｒｔｈｅｎｏｎｉｎｖａｓｉｖｅａｓｓｅｓｍｅｎｔｏｆｂｏｔｈｎｏｒｍａｌａｎｄａｂｎｏｒｍａｌｌｉｖｅｒｖａｓｃｕｌａｒｉｔｙｈａｖｅｏｆｅｒｅｄａｓｉｇｎｉｆｉｃａｎｔ...

Inhibitory effects of two oligosaccharideson murine melanoma experimental liver metastasis

ＩｎｈｉｂｉｔｏｒｙｅｆｅｃｔｓｏｆｔｗｏｏｌｉｇｏｓａｃｃｈａｒｉｄｅｓｏｎｍｕｒｉｎｅｍｅｌａｎｏｍａｅｘｐｅｒｉｍｅｎｔａｌｌｉｖｅｒｍｅｔａｓｔａｓｉｓＬＩＵＹｉＰｉｎｇ１，ＺＨＯＵＲｏｕＬｉ２，ＷＡＮＧＹｏｎｇＦｕ３ａｎｄＣＡＩＭｅ...

Iodine-125 interstitial brachytherapy for experimental liver cancer

Objective:To study the effect of iodine-125 interstitial brachytherapy on liver cancer. Methods: Animal model of human liver cancer was established by injecting SMMC-7721 cells cultivated in vitro subcutaneously into the flank of BALB/c nude mice. Nude mice with tumor of 5 mm in diameter were randomly divided into 2 groups (n = 10). One iodine-125 seed of apparent activity 0. 8 mCi was implanted into the center of tumor in treatment group, whereas an inactive seed was implanted in control group. The other 20 nude mice with tumor reaching 10 mm in diameter were also treated as above. The size of tumor was determined weekly after implantation, and pathological examination and blood routine were taken on the 28th day. Results: Tumor growth was obviously inhibited in treatment group of tumor of 5 mm in diameter, and there was statistically significant difference in tumor volume between treatment and control groups (P<0. 01). Around iodine-125 seed, apparent necrosis of tumor was shown in treatment group, accompanied by karyopyknosis and reduced plasma in residual tumor cells microscopically. Tumor growth was not inhibited in either treatment or control group of tumor of 10 mm in diameter. There was no obvious adverse effect except for decreased white blood cells in treatment groups. Conclusion: There is certain effect of iodine-125 interstitial brachytherapy on liver cancer, which is associated with the size of tumor.

Immunohistochemical study on expression of epidermal growth factor receptor at hepatocyte nuclei in experimental rat liver cirrhosis

ＩｍｍｕｎｏｈｉｓｔｏｃｈｅｍｉｃａｌｓｔｕｄｙｏｎｅｘｐｒｅｓｓｉｏｎｏｆｅｐｉｄｅｒｍａｌｇｒｏｗｔｈｆａｃｔｏｒｒｅｃｅｐｔｏｒａｔｈｅｐａｔｏｃｙｔｅｎｕｃｌｅｉｉｎｅｘｐｅｒｉｍｅｎｔａｌｒａｔｌｉｖｅｒｃｉｒｈｏｓｉｓＹＡＮＪｉｎＰｉｎ...

DYNAMIC CHANGES OF ITO CELLS IN EXPERIMENTAL LIVER CIRRHOSIS OF RAT

That Ho cells in rat liver express desmin was confirmed by immunohistochemical technique. Consequently, changes of desmin-positive cells, lysozyme-positive cells and fibronectin were further studied in experimental cirrhosis of rat. It was found that desmln- positive cells, with the transitional feature between Ito cells and myofibroblasts or fibrobiasts under electron microscope, increased in number and expression of desmin in the necrotic areas as well as in the cellular fibrous septa, but decreased in number in the fibrous septa except those areas closed to the edges of the septa. These results suggested that Ito cells, myofibroblasts and fibroblasts might belong to the same cellular system and play an important role in the pathogenesis of cirrhosis. Meanwhile, it was also noted that changes of both fibronectin and lysozymepositive cells were correlated with those of desmin-positive cells. These provide evidence in vivo that flbronectin and Kupffer cells might exert certain effects on the migration and proliferation of Ito cells in liver cirrhosis.

An experimental and clinical study of liver fibrosis

ＡｎｅｘｐｅｒｉｍｅｎｔａｌａｎｄｃｌｉｎｉｃａｌｓｔｕｄｙｏｆｌｉｖｅｒｆｉｂｒｏｓｉｓＫｏｎｇＸｉａｎｔａｏ（孔宪涛），ＧａｏＦｅｎｇ（高锋）（ＤｅｐａｒｔｍｅｎｔｏｆＣｌｉｎｉｃａｌＩｍｍｕｎｏｌｏｇｙ，ＣｈａｎｇｚｈｅｎｇＨｏｓｐｉｔａｌ，Ｓ...

Bone disorders in experimentally induced liver disease in growing rats

AIM: To investigate the change of bone parameters in a new model of experimentally induced liver cirrhosis and hepatocellular carcinoma (HCC) in growing rats.METHODS: Fischer-344 rats (n = 55) were used. Carbon tetrachloride (CCl4), phenobarbital (PB), and a single diethylnitrosamine (DEN) injection were used. Animals were killed at wk 8 and 16. Bone mineral content, femoral length, cortical index (quotient of cortical thickness and whole diameter) and ultimate bending load (Fmax)of the femora were determined. The results in animals treated with DEN+PB+CCl4 (DPC, n = 21) were compared to those in untreated animals (UNT,n = 14) and in control group treated only with DEN+PB (DP, n = 20).RESULTS: Fatty liver and cirrhosis developed in each DPC-treated rat at wk 8 and HCC was presented at wk 16. No skeletal changes were found in this group at wk 8,but each parameter was lower (P＜0.05 for each) at wk 16 in comparison to the control group. Neither fatty liver nor cirrhosis was observed in DP-treated animals at any time point. Femoral length and Fmax values were higher (P＜0.05 for both) in DP-treated animals at wk 8 compared to the UNT controls. However, no difference was found at wk 16.CONCLUSION: Experimental liver cirrhosis and HCC are accompanied with inhibited skeletal growth, reduced bone mass, and decreased mechanical resistance in growing rats. Our results are in concordance with the data of other studies using different animal models. A novel finding is the transiently accelerated skeletal growth and bone strength after a 8-wk long phenobarbital treatment following diethylnitrosamine injection.

Recurrence or metastasis of HCC:predictors,early detection and experimental antiangiogenic therapy

AIM To investigate the predictors forrecurrence or metastasis of HCC,and toevaluate the effect of antiangiogenic therapy onthe growth of transplantable human HCC in nudemice.METHODS RT-PCR was used to measure theexpression of matrix metalloproteinase-9(MMP-9)and vascular endothelial growth factor(VEGF)in 56 pairs of nontumorous liver andtumor samples.Sixty blood samples from humanHCC were examined by nested RT-PCR to find outAFP mRNA.Recombinant human endostatin andpolyclonal antibody against VEGF wereadministered to treat human HCC transplanted innude mice.RESULTS Thirty of 56 HCC samples showedstronger expression of MMP-9 in tumoroustissues than in nontumorous tissues.Fifteen ofthe 26 patients with relative expression level ofMMP-9 more than 0.34 developed tumorrecurrence or metastasis,whereas only 7 of 30patients with relative expression level less than0.34 developed tumor recurrence(P【0.05).There was no significant difference in therelative expression level of VEGF betweenpatients with postoperative recurrence ormetastasis and those without recurrence.AFPmRNA was detectable in 53.3% of patients withHCC.The sensitivity and specificity of AFPmRNA as a marker to detect hematogenousdissemination of HCC cells was 81.8% and84.4%,respectively.Recombinant human endostatin and polyclonal antibody against VEGFinhibited the growth of transplantable HCC innude mice by 52.2% and 45.7%,respectively.CONCLUSION MMP-9 expression in HCCcorrelates with the postoperative recurrence ormetastasis of HCC.Patients with high level ofMMP-9 expression in HCC are susceptible tometastasis.AFP mRNA could serve as anindicator of hematogenous spreading of HCCcells in circulation and a predictor of recurrenceor metastasis of HCC.Antiangiogenesis may bean adjuvant therapy for HCC.

Clinical and experimental study on regional administration of phosphorus32 glassmicrospheres in treating hepatic carcinoma

AIM To study the therapeutical effectiveness, dosage range and toxic adverse effects of domestic phosphorus 32 glass microsphere and evaluate its clinical significance. METHODS Ⅰ. Fifty two BALB/*!c tumor bearing male nude mice were allocated into treatment group( n =38) and control group( n =14). In the former group different doses of 32 P GMS were injected into the tumor mass, while in the latter 31 P GMS or no treatment was given. The experimental animals were sacrificed in batches, and then the tumors and their nearby tissues were examined by light and electron microscopy. Ⅱ. Through selective catheterization of hepatic artery, 32 P GMS was infused to 5 healthy domestic pigs in a dosage equivalent to the therapeutic dose for human being, and 31 P GMS was infused to another 5 healthy domestic pigs. Two pigs infused with contrast medium served as whole course blank controls. One pig from each group was surrendered to euthanasia at week 1, 4, 8 and 16 respectively. The ultrastructural histopath ological changes in liver tissues taken from different sites were evaluated semiquan titatively. Ⅲ. One hundred and twenty seven times of 32 P GMS intrahepatic artery interventional therapies were performed on 93 patients with hepatic carcinoma, including 79 cases of primary hepatic carcinoma and 14 cases of secondary hepatic carcinoma. 32 P GMS ( n =30), and group B, 32 P GMS and half dose of trans hepatic artery embolization (TAE) ( n =49) , and 18 patients with HCC by TAE only as control group C. Fourteen patients with secondary hepatic carcinoma were treated in the same way as group B or C. RESULTS Ⅰ. Comparing with the control group, the treatment group of tumor bearing nude mice attained the tumor inhibition rates of 59 7%-93 7% ( F =579 62, P <0 01) at 14*!d . At an absorbed dose of 7320Gy, the tumor cells were completely destroyed. When the absorbed doses ranged from 1830Gy to 3660Gy, most of the tumor cells showed the evidences of injury or necrosis, but there appeared some well differentiated tumor cells and enhanced effect of the autoimmunocytes. At an absorbed dose of 366Gy or less, some tumor cells still remained active proliferative ability. The definite anticancer effect appeared as early as 3d after intratumoral injection of 32 P GMS. Ⅱ. The cumulative amount of 32 P GMS in the target tissue after trans hepatic artery instillation attained more than 90% of the total dose administrated. Semiquantitative analysis of ultrastructral morphology in the experimental group showed no statistical difference between the nuclear abnormality (N abn ) and mitochondrial variability (M var ) at week 1 or 2, but revealed prominent difference (χ 2=6 70-9 68, P <0 01 , χ 2=65 09-115 09, P <0 001 ) as compared with those in the other groups. In the experimental group the N abn in tissues showed no significant difference between week 8 and week 16. No apparent changes were found in the stomach, spleen, kidney and lung tissues of the experimental pigs. Ⅲ. The therapeutical results of HCC patients in group A were closely approximated to those of group C, no hematological toxic side effects were noted, and the systemic reaction was mild. In some patients 2*!mos - 3*!mos after treatment some secondary foci appeared around the periphery of the primary lesion. In general better effectiveness was obtained in patients with small lesion. After analyzing by RIDIT method, the therapeutic result in group B was significantly better than that in group C, and secondary foci around the original lesion were rarely seen at 3*!mos after treatment. In group C the collateral circulation was reestablished along the periphery of primary foci and the secondary foci appeared more frequently, and required to undergo several courses of treatment. In group B, 4 cases of HCC were treated surgically as their mass decreased in size after 32 P GMS treatment.

Kupffer cell and apoptosis in experimental HCC

INTRODUCTION Our previous study has proved that Kupffer cellsmay have an inhibitory effect on the process ofhepatocarcinogenesis,however,their inhibitorymechanism needs exploring deeply.We performed acomparative study on the expression of PCNA,Bax,P53 and apoptosis of liver cancer cells usingimmunohistochemical technology and terminaldeoxynucleotidyl transferase (TdT)-mediateddUTP-digoxigenin nick end labeling(TUNEL)

Changing trends of surgical treatment of hilar bile duct cancer:clinical and experimental perspectives

HISTORICAL POINT OF VIEWCarcinoma of the confluence of the hepatic ductshas been thought to be a rare disease until recently.Carcinoma of the large hepatic ducts was rarelydiagnosed correctly premorterned.Because of itsdeeply seated location,resection was once thoughtto be impossible.In 1957,Altemeier reported

Thrombospondin-1 expression correlates with angiogenesis in experimental cirrhosis

AIM:To investigate the significance of Thrombospondin-1 (TSP-1) expression and its relationship with angiogenesis during experimental fibrosis. METHODS:Cirrhosis was induced in male Wistar rats by intraperitoneal administration of diethyl nitrosamine (DEN). The serial sections from liver tissues were stained with anti-CD34 and anti-TSP-1 antibodies before being quantitated by light microscopy. RESULTS:Our results showed that of TSP-1 expression gradually increases according to the severity of fibrosis (GroupⅠvs group Ⅱ, Group Ⅲ and Group Ⅳ;Group Ⅱ vs group Ⅲ and group Ⅳ;group Ⅲ vs group Ⅳ, P < 0.05). Moreover, TSP-1 expression was found to be correlated with angiogenesis (P < 0.05). CONCLUSION:The correlative evidence of the link between TSP-1 and fibrosis or angiogenesis provided by this study suggests that besides its role as a strong promoter of transforming growth factor-β1 (TGF-β1), TSP-1 might have an additional role in liver fibrogenesis by stimulating angiogenesis and this protein could be a potential target to prevent fibrogenesis in chronic inflammatory diseases of the liver.

Free radicals in development of experimental gastric carcinoma and precan cerous lesions in ducedby N-methyl-N'-nitro-N-nitrosoguanidine in rats

ＦｒｅｅｒａｄｉｃａｌｓｉｎｄｅｖｅｌｏｐｍｅｎｔｏｆｅｘｐｅｒｉｍｅｎｔａｌｇａｓｔｒｉｃｃａｒｃｉｎｏｍａａｎｄｐｒｅｃａｎｃｅｒｏｕｓｌｅｓｉｏｎｓｉｎｄｕｃｅｄｂｙＮｍｅｔｈｙｌＮ＇ｎｉｔｒｏＮｎｉｔｒｏｓｏｇｕａｎｉｄｉｎｅｉｎ...

Hepatitis C Virus Experimental Model Systems and Antiviral drug Research

An estimated 130 million people worldwide are chronically infected with hepatitis C virus (HCV) making it a leading cause of liver disease worldwide. Because the currently available therapy of pegylated interferon-alpha and ribavirin is only effective in a subset of patients, the development of new HCV antivirals is a healthcare imperative. This review discusses the experimental models available for HCV antiviral drug research, recent advances in HCV antiviral drug development, as well as active research being pursued to facilitate development of new HCV-specific therapeutics.

Regulating effect of Chinese herbal medicine on the peritoneal lymphatic stomata in enhancing ascites absorption of experimental hepatofibrotic mice

AIM: To observe the regulatory effect of Chinese herbalmedicine on peritoneal lymphatic stomata and itssignificance in treating ascites in liver fibrosis model mice.METHODS: Two Chinese herbal composite prescriptionswere used separately to treat the carbon tetrachloride-induced mouse model of liver fibrosis. The histo-pathologicchanges of the liver sections (HE and VG stainings) wereobserved. The peritoneal lymphatic stomata was detected byscanning electron microscopy and computer imageprocessing. The changes of urinary volume and sodium ionconcentration were measured.RESULTS: In the model group, lots of fibrous tissue formedin liver and extended into the hepatic Iobulss to separatethem incompletely. In the treated and prevention groups,the histo-pathologic changes of liver was rather milder, onlyshowed much less fibrous tissue proliferation in the hepaticIobules. The peritoneal lymphatic stomata enlarged withincreased density in the experimental groups (diameter:PA, 3.07±0.69μm; PB, 2.82±0.37μm; TA, 3.25±0.82μmand TB, 2.82±0.56μm; density: PA, 7.11± 1.90 stomata@1000μm-2; PB, 8.76± 1.45 stomata@ 1000μm-2; TA, 6.55± 1.44 stomata@ 1000μm-2 and TB, 8.76 ± 1. 79 stomata@ 1000μm-2), as compared with the model group (diameter: 2.00 ±0.52μm; density: 4.45 ± 1. 05 stomata@ 1000 μm-2 ). Aftertreatment, the urinary volume and sodium ion excretionincreased in the experimental groupe ( PA, 231.28 ± 41. 09mmol@L-1; PB, 171.69± 27.48 mmol@L-1 and TA, 231.44±34.12 retool@ L-1 ), which were significantly different with thosein the model group (129.33 ± 36.75 rnmol@ L-1 ).CONCLUSION: Chinese herbal medicine has marked effectsin alleviating liver fibrosis, regulating peritoneal lymphaticstomata, improving the drainage of ascites from peritonealcavity and causing increase of urinary volume and sodiumion excretion to reduce the water and sodium retention, andthus have favorable therapeutic effect in treating ascites.

History,ethics,advantages and limitations of experimental models for hepatic ablation

Numerous techniques developed in medicine require careful evaluation to determine their indications,limitations and potential side effects prior to their clinical use.At present this generally involves the use of animal models which is undesirable from an ethical standpoint,requires complex and time-consuming authorization,and is very expensive.This process is exemplified in the development of hepatic ablation techniques,starting experiments on explanted livers and progressing to safety and efficacy studies in living animals prior to clinical studies.The two main approaches used are ex vivo isolated non-perfused liver models and in vivo animal models.Ex vivo non perfused models are less expensive,easier to obtain but not suitable to study the heat sink effect or experiments requiring several hours.In vivo animal models closely resemble clinical subjects but often are expensive and have small sample sizes due to ethical guidelines.Isolated perfused ex vivo liver models have been used to study drug toxicity,liver failure,organ transplantation and hepatic ablation and combine advantages of both previous models.

Liver transplantation as an alternative for the treatment of perihilar cholangiocarcinoma: A critical review

Background:Perihilar cholangiocarcinoma(phCCC)is a dismal malignancy.There is no consensus regard-ing the best treatment for patients with unresectable phCCC.The present review aimed to gather the current pieces of evidence for liver transplantation and liver resection as a treatment for phCCC and to build better guidance for clinical practice.Data sources:The search was conducted in PubMed,Embase,Cochrane,and LILACS.The related references were searched manually.Inclusion criteria were:reports in English or Portuguese literature that a)patients with confirmed diagnosis of phCCC;b)patients treated with a curative intent;c)patients with the outcomes of liver resection and liver transplantation.Case reports,reviews,letters,editorials,conference abstracts and papers with full-text unavailability were excluded from the analysis.Results:Most of the current literature is based on observational retrospective studies with low grades of evidence.Liver resection has better long-term outcomes than systemic chemotherapy or palliation ther-apy and liver transplantation is a good alternative for selected patients with unresectable phCCC.All candidates for resection or transplantation should be medically fit and free of intrahepatic or extrahep-atic diseases.As a general rule,patients presenting with a tumor having a longitudinal size>3 cm or extending below the cystic duct,lymph node disease,confirmed extrahepatic dissemination;intraoper-atively diagnosed metastatic disease;a history of other malignancies within the last five years,and did not complete chemoradiation regimen and were medically unfit should not be considered for transplan-tation.Some of these criteria should be individually assessed.Liver transplantation or resection should only be considered in highly experienced hepatobiliary centers,and any decision-making must be based on a multidisciplinary evaluation.Conclusions:phCCC is a complex condition with high morbidity.Surgical therapies,including hepatec-tomy and liver transplantation,are the best option for better long-term disease-free survival.