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Objectives: The prevention and treatment of liver transplant rejection remain challenging. We investigated the pathophysiological mechanisms of liver transplant rejection in rats and screened candidate genes to determine their degree of rejection response for possible development of potential therapeutic targets., Materials and Methods: Brown Norway-Brown Norway transplant tolerant models and Lewis-Brown Norway transplant rejection models were established. We collected liver tissue and venous blood at 7 days posttransplant for hematoxylin and eosin staining and RNA sequencing analysis, respectively. We conducted differential expression gene analysis, KEGG and GO enrichment analysis. We performed immunohistochemistry to detect highly expressed immunerelated proteins, including lymphocyte-specific protein tyrosine kinase, linker for activation of T cells, and 70-kDa T-cell receptor zeta-chain-associated protein kinase., Results: Significant differences were found in liver function and Banff scores between rejection and tolerant groups, indicating the successful establishment of liver transplant models. RNA-sequencing screened 7521 differentially expressed genes, with 3355 upregulated and 3058 downregulated. KEGG analysis of upregulated genes showed that 8 of the top 20 enrichment pathways were associated with immune system processes and 5 were related to immune system diseases. Among these immune pathways, 289 genes were upregulated; of these, 147 genes were removed after comparison with the IMMPORT database, of which 97 genes were significantly changed. Our GO analysis showed upregulated genes mainly participating in immune response processes, with downregulated genes mainly participating in metabolic processes. Real-time polymerase chain reaction and immunohistochemistry verified expression of the immune-related proteins, consistent with RNAsequencing results, which were mainly expressed in inflammatory cells in sinus and portal vein., Conclusions: Immune-related genes were found to be associated with liver transplant rejection. The 3 immune-related genes that we analyzed may play a role in liver transplant rejection and can possibly serve as candidate markers for monitoring the degree of liver transplant rejection.

Objectives: Orthotopic liver transplant remains technically challenging., Materials and Methods: We performed whole graft orthotopic liver transplants with different anhepatic times (≤20 min, n = 19; vs 30 min, n = 9) and partial orthotopic liver transplants in rats including a male-to-male Sprague-Dawley group (n = 15), a male-to-male Lewis-to-Brown Norway group (n = 20), and a male-to-male Sprague-Dawley-to-Lewis group (n = 20); there was also a female-to-male SpragueDawley group (n = 19)., Results: For the groups with ≤20-minute or 30-minute anhepatic time, 14-day and 30-day survival rates were 94.7%, 89.5%, 88.9%, and 88.9%, respectively, and there was no difference in survival (P = .716). For 50% orthotopic liver transplants from the male-tomale Sprague-Dawley group, 14-day and 30-day survival rates were 93.3% and 86.7%, respectively, with no difference between whole and 50% graft orthotopic liver transplant. The 14-day and 30-day survival rates were, respectively, 30% and 10% for the Lewis-to-Brown Norway group and 30% and 6.6% for the Sprague-Dawley-to-Lewis group, with no differences between the 2 groups (P = .564). Most of the recipient rats died within 72 hours. Acute rejections and wound dehiscence were the causes of death. Recipients from the female-to-male SpragueDawley orthotopic liver transplant group died shortly after surgery., Conclusions: Orthotopic liver transplants can be performed to achieve high success rates in the extended anhepatic time; however, orthotopic liver transplants from female Sprague-Dawley donor rats have a high risk of failure.

Although there is some progress in immunosuppressive therapy of acute rejection, there is still a lack of standardized diagnosis and treatment. For the acute rejection after liver transplantation, there is still a lack of an exact treatment at this stage. Tacrolimus (TAC) side effects will also affect the survival rate and quality of life of recipients after transplantation to a large extent. Rat orthotopic liver transplantation model was established and divided into three groups. In the tolerance group, Brown Norway (BN) to Lewis liver transplantation was used; in the rejection group, Lewis to BN liver transplantation was used; in the TAC group, TAC was injected after operation on the basis of establishing rejection model. The expression of GITRL in Kupffer cells and the change of cytokines were detected 7 days after operation. In this study, the animal model of acute rejection of rat liver transplantation was established to simulate the clinical allogeneic liver transplantation, and the important role of TAC in the acute rejection of rat liver transplantation was evaluated.

ABO-incompatible (ABO-i) living-donor liver transplantation (LDLT) is performed if an ABO-compatible graft cannot be obtained. However, a perfect desensitization protocol has not been established worldwide, especially for simultaneous ABO-i LDLT and splenectomy. We herein report two cases of ABO-i LDLT. To the best of our knowledge, this is the first case report of ABO-i LDLT in an adult patient in China. Splenectomy and T-cell-targeted immunosuppression (basiliximab) was used to overcome the blood group barrier in these recipients. The patients had good graft function without signs of antibody-mediated rejection throughout the 12-month follow-up. Thus, ABO-i LDLT with splenectomy is undoubtedly life-saving when an ABO-compatible graft cannot be obtained for patients in critical condition.

Objective To explore the classification and changes of macrophage subsets in liver transplant rejection. Methods Rat liver transplantation model were established and divided into immune tolerance group (B-B), where the liver of BN rat donors was transplanted to BN rat recipients, and immune rejection group (L-B), in which the liver of Lewis rat donors was transplanted to BN rat recipients. Single-cell RNA sequencing and highthroughput RNA sequencing were used to distinguish the macrophage subsets of rat liver transplantation, and to find differential gene in rejection reactions. Immunohistochemistry was used to determine the changes and distribution of protein expression and cell subsets. Results CD68 positive macrophages were higher in the rejection group than that in the tolerance group (P < 0.05), and macrophages could be divided into 9 subsets. During the rejection reaction, the CXC chemokine ligand 9 (CXCL9) in the 8th subsets of macrophages was significantly increased, while the gene for white blood cell differentiation antigen 74 (CD74) in the 5th subsets was significantly increased (P < 0.05). CD74 ranked first in the differential gene synthesis of macrophages during rejection, followed by CXCL9. Compared with the tolerance group, a large number of CD74 positive macrophages were observed in the hepatic portal area of the rejection group, and the infiltration of CD74 positive macrophages in the hepatic sinuses was also significantly increased (P < 0.05), while a large number of CXCL9 positive macrophages were observed in the hepatic portal area and hepatic sinuses of the rejection group, especially in the portal area (P < 0.05), and CD14 positive cells were significantly increased (P < 0.05). Conclusions The CD74 positive macrophage subsets and CXCL9 positive macrophage subsets may be key subgroups in promoting liver transplant rejection, improving the mechanism of macrophage action in liver transplant rejection. [ABSTRACT FROM AUTHOR]

Objectives: T‐cell immunoglobulin domain and mucin domain‐4 (TIM‐4) is selectively expressed on antigen‐presenting cells (APCs) and modulates various immune responses. However, the role of TIM‐4 expressed by Kupffer cells (KCs) in liver fibrosis remains unclear. The present study aimed to explore whether and how TIM‐4 expressed by KCs is involved in liver fibrosis. Materials and Methods: Mice chronic liver fibrosis models were established and divided into the olive‐induced control group, CCL4‐induced control group, olive‐induced TIM‐4 interference group and CCL4‐induced TIM‐4 interference group. Different techniques were used to monitor the fibrotic effects of TIM‐4, including histopathological assays, Western blotting, ELISA and transmission electron microscopy. Additionally, mice liver transplant models were established to determine the fibrotic effects of TIM‐4 on fibrosis after liver transplantation (LT). Results: We found that the induction of liver fibrosis by CCL4 was associated with TIM‐4 expression in KCs. TIM‐4 interference essentially contributed to liver fibrosis resolution. KCs from the TIM‐4 interference group had decreased levels of pro‐fibrotic markers, reduced TGF‐β1 secretion and inhibited hepatic stellate cell (HSC) differentiation into myofibroblast‐like cells. In addition, we used GdCl3 to verify that KCs are the primary source of TGF‐β1 during fibrosis progression. Moreover, KCs from CCL4‐induced mice showed increased ROS production, mitophagy activation and TGF‐β1 secretion. However, TIM‐4 interference in the KCs inhibited Akt1‐mediated ROS production, resulting in the suppression of PINK1, Parkin and LC3‐II/I activation and the reduction of TGF‐β1 secretion during liver fibrosis. Additionally, TIM‐4 interference potentially attenuated development of fibrosis after LT. Conclusions: Our findings revealed the underlying mechanisms of TIM‐4 interference in KCs to mitigate liver fibrosis. [ABSTRACT FROM AUTHOR]