Age-related changes in metabolites in young donor livers and old recipient sera after liver transplantation from young to old rats.

Liver ageing not only damages liver function but also harms systemic metabolism. To better understand the mechanisms underlying liver ageing, we transplanted the livers of young rats to young and old rats and performed untargeted metabolomics to detect changes in the metabolites in the liver tissues and sera. A total of 153 metabolites in the livers and 83 metabolites in the sera were different between the old and young rats that did not undergo liver transplantation; among these metabolites, 7 different metabolites were observed in both the livers and sera. Five weeks after liver transplantation, the levels of 25 metabolites in the young donor livers were similar to those in the old rats, and this result probably occurred due to the effect of the whole-body environment of the older recipients on the young livers. The 25 altered metabolites included organic acids and derivatives, lipids and lipid-like molecules, etc. In the sera, the differences in 78 metabolites, which were significant between the young and old rats, were insignificant in the old recipient rats and made the metabolic profile of the old recipients more similar to that of the young recipients. Finally, combining the above metabolomic data with the transcriptomic data from the GEO, we found that the altered metabolites and genes in the liver were enriched in 9 metabolic pathways, including glycerophospholipid, arachidonic acid, histidine and linoleate. Thus, this study revealed important age-related metabolites and potential pathways as well as the interaction between the liver and the whole-body environment.

A national report from China Liver Transplant Registry: steroid avoidance after liver transplantation for hepatocellular carcinoma.

OBJECTIVE: We aimed to evaluate the efficacy and safety of steroid-free immunosuppression after liver transplantation (LT) for hepatocellular carcinoma (HCC). METHODS: We retrospectively analyzed HCC recipients without steroids after LT (SF group, n=368) based on the China Liver Transplant Registry (CLTR) database. These recipients were matched 1:2 with patients using steroids (S group, n=736) for the same period after LT for HCC, according to propensity scores. RESULTS: Multivariate analysis indicates that recipients with younger age [odds ratio (OR), 1.053; P=0.011], preoperative hepatitis B virus (HBV) DNA ≥1,000 copies/mL (OR, 2.597; P=0.004) and beyond Milan criteria (OR, 4.255; P<0.001) were identified as the risk factors associated with tumor recurrence in steroid avoidance recipients after LT. The patients fulfilling the Milan criteria in the SF group presented higher overall and tumor-free survival rates than those in the S group (P<0.05). Multivariate analysis revealed that recipient beyond Milan criteria was an independent prognostic factor for overall survival (OR, 1.690; P<0.001) and tumor-free survival (OR, 2.066; P<0.001). The incidences of new-onset diabetes mellitus (21.20%vs. 33.29%, P<0.001), new-onset hypertension (10.05%vs. 18.61%, P<0.001) and hyperlipidemia (4.08%vs. 7.20%, P=0.042) were significantly lower in the SF group. CONCLUSIONS: Steroid-free immunosuppression could be safe and feasible for HBV-related HCC patients in LT. Age, HBV DNA level and Milan criteria maybe risk factors associated with tumor recurrence in steroid avoidance recipients. Recipient beyond Milan criteria was an independent prognostic factor and recipient fulfilling Milan criteria can benefit the most from steroid-free immunosuppression.

In vivo therapeutic potential of Inula racemosa in hepatic ischemia-reperfusion injury following orthotopic liver transplantation in male albino rats.

Hepatic ischemia-reperfusion (I/R) injury mainly occurs following hepatic resection and liver transplantation and cause severe liver damage, organ injuries, and dysfunction. Pro-inflammatory cytokines that promote injury are released when kupffer cell activates after getting induced by I/R. Repercussions of oxidative stress and cardiac function against isoproterenol based myocardial infarction are caused by flavonol glycosides which are found in high concentrations in Inula racemosa (Ir).The root was deemed to have analgesic and anti-inflammatory effects, and no report has been published about the liver-protective activity against hepatic I/R. Therefore, the present study was aimed to understand the therapeutic impact of Ir in hepatic I/R injury. Male albino, Wistar strain rats were used and were grouped into four total phenolic content, free radical scavenging activity and serum enzymes were determined. Histopathological and immunohistochemical analysis were also carried out. Inflammatory cytokines such as tumor necrosis factor-alpha (TNF-α) and interleukin (IL-6) and protein expression of p53, bax, and bcl-2 were determined. The administration of extracts of Ir significantly increased total phenolic and free radical scavenging activity. Altered cellular morphology, cytokines and aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were returned to near normal level. IL-6 and TNF-α levels were reduced more than 25% following treatment. Also, the protein expression of p53, bax, and bcl-2 were also returned to near normal level. Taking all these data together, it is suggested that the extracts of Ir may be a potential therapeutic agent for providing several beneficial effects in hepatic I/R injury.

Expansion of the Milan criteria without any sacrifice: combination of the Hangzhou criteria with the pre-transplant platelet-to-lymphocyte ratio.

BACKGROUND: The Hangzhou criteria expand the Milan criteria safely and effectively in selecting hepatocellular carcinoma (HCC) candidates for liver transplantation (LT), but some patients exceeding the Milan but fulfilling the Hangzhou criteria still show poor outcomes due to early tumor recurrence. In this study, the platelet-to-lymphocyte ratio (PLR) was employed to differentiate high-risk tumor recurrence recipients, and a new method combining PLR and the Hangzhou criteria was established. METHODS: The clinical data of 343 LT for HCC were retrospectively analyzed. Receiver operating characteristic (ROC) analysis was used to determine the PLR cut-off value to stratify patients exceeding the Milan but fulfilling the Hangzhou criteria. The recurrence-free survival (RFS) of recipients was compared after stratification. The Hangzhou criteria & PLR method was proposed and its feasibility was validated by ROC analysis. RESULTS: PLR 120 was the most significant cut-off value when comparing RFS of patients exceeding the Milan but fulfilling the Hangzhou criteria. After stratification, the 1-, 3-, and 5-year RFS of patients exceeding the Milan but fulfilling the Hangzhou criteria with PLR < 120 were 84.2%, 73.3%, and 73.3%, respectively, comparable with 85.7%, 73.9%, and 72.8%, respectively, in patients fulfilling the Milan criteria (P = 0.885). Patients exceeding the Milan but fulfilling the Hangzhou criteria with PLR ≥ 120 showed poor outcomes, which were similar in patients exceeding the Hangzhou criteria; 1-, 3-, and 5-year RFS were only 37.5%, 12.5%, and 12.5% vs. 32.3%, 17.6%, and 15.1%, respectively (P = 0.887). ROC analysis demonstrated that the ROC area of the Hangzhou criteria & PLR method was 0.768 for RFS. Multivariate analysis confirmed that PLR ≥ 120 was independently associated with RFS of patients exceeding the Milan but fulfilling the Hangzhou criteria. CONCLUSIONS: The Hangzhou criteria combined with the pre-transplant PLR can accurately exclude high-risk tumor recurrence recipients; this approach expands the Milan criteria effectively without any sacrifice.

Meta-Analysis of Laparoscopic versus Open Hepatectomy for Live Liver Donors.

OBJECTIVE: To document the safety and efficacy of laparoscopic living donor hepatectomy in comparison with open liver resection for living donor liver transplantation. METHODS: Medline database, EMASE and Cochrane library were searched for original studies comparing laparoscopic living donor hepatectomy (LLDH) and open living donor hepatectomy (OLDH) by January 2015. Meta-analysis was performed to evaluate donors' perioperative outcomes. RESULTS: Nine studies met selection criteria, involving 1346 donors of whom 318 underwent LLDH and 1028 underwent OLDH. The Meta analysis demonstrated that LLDH group had less operative blood loss [patients 1346; WMD: -56.09 mL; 95%CI: -100.28-(-11.90) mL; P = 0.01], shorter hospital stay [patients 737; WMD: -1.75 d; 95%CI: -3.01-(-0.48) d; P = 0.007] but longer operative time (patients 1346; WMD: 41.05 min; 95%CI: 1.91-80.19 min; P = 0.04), compared with OLDH group. There were no significant difference in other outcomes between LLDH and OLDH groups, including overall complication, bile leakage, postoperative bleeding, pulmonary complication, wound complication, time to dietary intake and period of analgesic use. CONCLUSIONS: LLDH appears to be a safe and effective option for LDLT. It improves donors' perioperative outcomes as compared with OLDH.

Post-Operative Complications in Living Liver Donors: A Single-Center Experience in China.

The gap between the growing demand for available organs and the cadaveric organs facilitates the adoption of living donor liver transplantation. We retrospectively identified and evaluated the post-operative complications as per the modified Clavien classification system in 152 living liver donors at at the First Affiliated Hospital, College of Medicine, Zhejiang University between December, 2006 and June, 2014. Post-operative complications were observed in 61 patients (40.1%) in the present study, but no mortality was reported. Complications developed in 58 (40.0%) right, 1 (33.3%) left, and 2 (66.7%) lateral left hepatectomy donors. The prevalence of re-operation was 1.3%. Grade I and II complications were observed in 38 (25.0%) and 11 (7.2%) donors, respectively. Grade IIIa complications developed in 9 (5.9%) donors and only 3 (2.0%) patients reported grade IIIb complications. The most common complication was pleural effusion that occurred in 31 (20.4%) donors. No significant prognostic baseline factor was identified in this study. In conclusion, living donors experienced various complications, which were usually mild and had a good prognosis.

Rapamycin and tacrolimus differentially modulate acute graft-versus-host disease in rats after liver transplantation.

Acute graft-versus-host disease (aGVHD) is a serious complication of liver transplantation (LTx); it occurs in 1% to 2% of liver allograft recipients. The condition has a poor prognosis and poses major diagnostic and therapeutic challenges. A rat model of aGVHD after LTx has been developed, and a relative decrease in regulatory T (Treg) cells has been shown to be associated with this model. Interest has been expressed in the effects of different immunosuppressive agents on CD4+CD25+Foxp3+ Treg cell homeostasis. Rats with aGVHD after LTx were treated with tacrolimus (FK506), rapamycin (RAPA), or no immunosuppressive drug. Those that received RAPA survived longer (91.4 + or - 8.1 days) than those in the FK506 group (62.3 + or - 13.4 days) or the control group (22.9 + or - 1.2 days). Flow cytometry analysis showed that Treg cells, as a percentage of peripheral blood mononuclear cells (PBMCs), were more abundant in the RAPA group (6.8% + or - 0.8%) than in the FK506 group (1.7% + or - 0.4%) or the control group (2.0% + or - 0.4%). Immunohistochemistry demonstrated more Foxp3+ staining of intestinal cells in the RAPA group than in the FK506 group or the control group. In conclusion, the reduced mortality induced by RAPA in a rat model of aGVHD after LTx was associated with higher percentages of CD4+CD25+Foxp3+ Treg cells in PBMCs in blood and tissues than those occurring after the administration of FK506.

A pathogenic role of IL- 17 at the early stage of corneal allograft rejection.

PURPOSE: Th17, recently identified as a new subset of effector Th cells, has been shown to be involved in microbe infection and autoimmunity. However, the role of these cells in organ allograft rejection remains largely unknown. In this study, we investigate whether Th17 cells participate in allogeneic corneal rejection in a mouse model. METHODS: Donor cornea (C57BL/6) was transplanted into orthotopic graft bed of Balb/c recipients. At different time points after keratoplasty, the expression of Th17 and Th1- related cytokines in draining cervical lymph nodes (LN) and grafted cornea was examined by flow cytometry and quantitative RT- PCR, respectively. Furthermore, IL- 17(-/-) Balb/c mice were used to determine the effects of Th17 cells on allogeneic cornea survival. Finally, the profiles of Th1 and proinflammatory cytokines in IL- 17(-/-) recipients after transplantation were examined. RESULTS: Th17 expression was enhanced significantly in inflamed transplants and draining lymph nodes at the early stage of allocorneal rejection, while upregulation of Th1 producing IFN- gamma was seen in the late phase. Upon activation by allogeneic accessory cells, responder cells in draining LN from transplanted recipients secreted high levels of IL- 6, TGF- beta and IL- 21 compared to controls, which may drive naive T cells to differentiate into Th17 cells. Importantly, IL- 17 deficiency led to the delayed development of allogeneic rejection, but did not affect the overall survival time of transplants. This effect correlated with restrained Th1 polarization and decreased production of proinflammatory cytokines. CONCLUSION: Th17 cells play a disease-promoting role at the early stage of corneal allograft rejection.

Correlation between ECT2 gene expression and methylation change of ECT2 promoter region in pancreatic cancer.

BACKGROUND: Pancreatic cancer is closely related to epigenetic abnormality. The epithelial cell transforming sequence 2 gene (ECT2) plays a critical role in Rho activation during cytokinesis, and thus may play a role in the pathogenesis of pancreatic cancer. In this study, we investigated the relationships between aberrant expression and epigenetic changes of the ECT2 gene in pancreatic cancer. METHODS: Four cell lines (PANC-1, Colo357, T3M-4 and PancTuI) and pancreatic ductal adenocarcinoma (PDAC) tissues were used for mRNA detection. After restriction isoschizomer endonucleases (MspI/HpaII) were used to digest the DNA sequence (5'-CCGG-3'), PCR was made to amplify the product. And RT-PCR was applied to determine the expression of the gene. RESULTS: The mRNA expression of the ECT2 gene was higher in pancreatic tumor tissue than in normal tissue. The gene was also expressed in the 4 PDAC cell lines. The methylation states of the upstream regions of the ECT2 gene were almost identical in normal, tumor pancreatic tissues, and the 4 PDAC cell lines. Some of the 5'-CCGG-3' areas in the upstream region of ECT2 were methylated, while others were unmethylated. CONCLUSIONS: The oncogene ECT2 is overexpressed in pancreatic tumor tissues as verified by RT-PCR detection. The methylation status of DNA in promoter areas is involved in the gene expression, along with other factors, in pancreatic cancer.

Epigenetic changes of pituitary tumor-derived transforming gene 1 in pancreatic cancer.

BACKGROUND: Pancreatic cancer is a devastating disease with abnormal genetic changes. The pituitary tumor-derived transforming gene (PTTG) is considered to be implicated in the tumorigenesis of cancers when the gene is epigenetically transformed. In this study, we investigated the relationships between aberrant expression and epigenetic changes of the PTTG1 gene in pancreatic cancer. METHODS: We chose 4 cell lines (PANC-1, Colo357, T3M-4 and PancTuI) and pancreatic ductal adenocarcinoma (PDAC) tissues. After using restriction isoschizomer endonucleases (MspI/HpaII) to digest the DNA sequence (5'-CCGG-3'), we performed PCR reaction to amplify the product. And RT-PCR was applied to determine the gene expression. RESULTS: The mRNA expression of the PTTG1 gene was higher in pancreatic tumor than in normal tissue. The gene was also expressed in the 4 PDAC cell lines. The methylation states of the upstream regions of the PTTG1 gene were almost identical in normal, tumor pancreatic tissues and the 4 PDAC cell lines. Some (5'-CCGG-3') areas in the upstream region of PTTG1 were methylated, while some others were unmethylated. CONCLUSIONS: The oncogene PTTG1 was overexpressed in pancreatic tumor tissues and verified by RT-PCR detection. The methylation status of DNA in promoter areas was involved in the gene expression with the help of other factors in pancreatic cancer.

Calcitriol prolongs recipient survival by inducing expression of zinc-finger protein A20 and inhibiting its downstream gene following rat orthotopic liver transplantation.

Calcitriol has important immunomodulation action and can prolong recipient survival after organ transplantation. The data in this study demonstrated that treatment of liver allograft recipient with calcitriol can protect allograft from acute rejection and prolong recipient's survival. Calcitriol inhibited expression of proinflammatory cytokine such as Interleukin-2 and Interferon-gamma intragraft. It also inhibited expression of nuclear factor kappaB (NF-kappaB) significantly as a result of enhancing its inhibitory protein I kappa B (IkappaB). As well, expression of zinc-finger protein A20 (A20) was enhanced significantly. The results suggest that calcitriol exerts its immunosuppression action in part through inducement of the A20, IkB, inhibition of NF-kB, and resultant proinflammatory expression pathway.