Ischemia-reperfusion injury (IRI) and subsequent rejection remain the paramount pathological barriers to long-term graft survival following liver transplantation. Traditionally viewed as mere ‘first responders’ in the acute inflammation of IRI, neutrophils are now recognized, based on recent advances, as pivotal regulators that bridge innate and adaptive immunity throughout the entire post-transplant course. This review aims to systematically delineate the dual pathological mechanisms of neutrophils in both IRI and rejection post-LT. During the initial phase of IRI, we focus on the robust activation of neutrophils, driven by damage-associated molecular patterns (DAMPs), with a particular emphasis on the formation of neutrophil extracellular traps (NETs). NETs act not only as key effectors causing sinusoidal microcirculatory dysfunction and direct hepatocellular injury, but their released histones and proteases also serve as potent danger signals, amplifying the local inflammatory cascade. The central thesis of this review is that the inflammatory microenvironment, orchestrated by neutrophils during IRI, provides the essential immunological substrate for subsequent rejection. We delve into the mechanisms by which neutrophils bridge to adaptive immunity: NETs serve as a scaffold for autoantigens, activating B cells and promoting the production of donor-specific antibodies (DSA), thereby driving antibody-mediated rejection (AMR). Concurrently, chemokines released by neutrophils efficiently recruit effector T cells and, through interactions with antigen-presenting cells, exacerbate cell-mediated rejection (CMR). Finally, we prospect future therapeutic directions, emphasizing that targeting common pathways within this ‘injury-immunity’ axis—such as inhibiting DAMP release, blocking NET formation, or employing pro-resolving mediators to actively terminate inflammation—represents a pivotal strategy to break the vicious cycle of IRI and rejection and achieve long-term immune tolerance.

Mucosal-associated invariant T (MAIT) cells, representing one of the most abundant subsets of unconventional T cells, have been shown to play a significant role in regulating immune responses. However, their immunoregulatory roles in the context of liver transplantation (LT) immunity remain largely undefined. To address this, we conducted single-cell RNA/TCR sequencing, flow cytometry, and multiplex immunohistochemical (mIHC) assays to identify the proportion and characteristics of CD8+ MAIT cells in humans and mice following liver transplantation. We found that CD8+ MAIT cells were prominently represented in the single-cell CD8 profiles of human transplanted livers, demonstrating strong signalling associations with macrophages, whilst the fractional populations of MAIT1 and MAIT17 were distinctly clustered. In parallel, the proportion of CD8+ MAIT cells was significantly elevated in mouse LT models, revealing a dynamic trend where percentages increased at 1 and 2 weeks post-transplant, peaking at 3 weeks. Furthermore, using established MR1 knockout (MR1KO) LT mice, we observed that mice lacking MAIT cells exhibited milder rejection responses, indicating that MR1 mediates rejection by influencing the remodelling of the TCR repertoire after transplantation. Collectively, our study reveals that MAIT cells play a critical role in LT rejection, as MR1KO alleviated inflammatory responses and mitigated rejection via TCR repertoire remodelling. By mapping the dynamic changes of MAIT cells throughout the rejection process, these findings lay the groundwork for further investigations into the role of these cells in transplant immunity.

Aim: Exosomes derived from adipose-derived stem cells (ASCs) in mice have been reported to influence immune regulation. Yet, the potential immunological effects of ASCs-derived exosomes and their interaction with lymphocytes during transplant immunity remain understudied. Methods: ASCs from BALB/c mice, along with their conditioned culture medium, were collected for the extraction, isolation, and comprehensive characterization of exosomes. Splenic cell suspensions were isolated from BALB/c mice and subsequently processed for downstream analyses. Lymphocytes were isolated via gradient centrifugation and stimulated in vitro with the purified exosomes to assess their functional responses. Lymphocyte proliferation was quantified using the CCK8 assay, and the relative frequencies of CD4+ T cells, CD8+ T cells, Treg cells, NK (natural killer) cells, macrophages, B cells, dendritic cells (DCs), and Th17 cells were determined through flow cytometric analysis. Before establishing the skin transplantation model, the mice were administered PBS, 0.5 × 108 exosomes, 1 × 108 exosomes, 1.5 × 108 exosomes, or ASCs via intravenous injection through the tail vein. Seven days after transplantation, the spleens, drainage lymph nodes, and blood samples were harvested for lymphocyte isolation and further downstream analyses. Results: Exosomes derived from ASCs significantly increased the CD4+/CD8+ ratio and Treg cell levels, without inducing any notable changes in Th17 cell content or CTLA-4 protein expression in CD4+ T cells. Compared to the PBS-treated group, both ASC and exosome treatment groups demonstrated an enhanced CD4+/CD8+ ratio, increased Treg cell content, and elevated CTLA-4 protein expression in spleen tissue following skin transplantation, while Th17 cell levels remained unaffected. Compared to the ASC treatment group, the exosome group exhibited a higher CD4+/CD8+ ratio and Treg cell levels, alongside a reduced proportion of PD-1+ Treg cells and lower CTLA-4 protein expression in CD3+CD4+ T cells. No significant differences were observed in the proportions of NK cells, macrophages, B cells, and DCs in the spleens across all treatment groups. In peripheral blood, an increased proportion of CD3+ T cells, macrophages, and DCs was detected, accompanied by a reduced proportion of NK cells and B cells. In the draining lymph nodes, no significant changes were observed in the proportions of CD3+ T cells and B cells, while macrophages, NK cells, and DCs showed elevated proportions. In the exosome-treated group, mouse grafts exhibited a disorganized and thinner granular layer, accompanied by focal regions of inflammatory cell infiltration. Both exosome and ASC treatments significantly extended the survival of skin grafts. Conclusion: Exosomes derived from ASCs promote lymphocyte proliferation and modulate their phenotypic profiles in mouse skin graft models, effectively extending graft survival.

Immune rejection poses a major challenge in organ transplantation, with tissue-resident memory T (TRM) cells playing a critical role in graft rejection. This study investigated the impact of CCR8 on TRM cells using single-cell RNA sequencing (scRNA-seq), flow cytometry, and immunohistochemistry. The results show that CCR8 expression was upregulated on CD8⁺ TRM cells after transplantation, peaking on day 7. Blocking or knocking out CCR8, as well as neutralizing CCL1 and CCL8, significantly reduced CD8⁺ TRM cell accumulation in the graft and their cytokine production. These treatments prolonged graft survival, alleviated rejection severity, and impaired the ability of CD8⁺ TRM cells to produce GZMB, IFN-γ, and IL-2. Single-cell analysis of skin transplantation revealed that loss of CCR8 disrupted macrophage-T cell interactions, particularly CD8⁺ TRM-macrophage crosstalk, while enhancing CD40, PD-L1, and NRG signaling. These findings suggest that targeting CCR8 to limit the accumulation and function of CD8⁺ TRM cells may be an effective strategy to alleviate transplant rejection.

BACKGROUNDS:Hepatic ischemia-reperfusion injury (IRI) triggers macrophage activation, which in turn mediates inflammatory responses and affects tissue repair and injury severity. Pentraxin 3 (PTX3) is vital in immune regulation and inflammatory processes. In this study, we aim to investigate the potential role of PTX3 in macrophage-mediated hepatic IRI. METHODS:Gene expression profiles and single-cell data were obtained from the Gene Expression Omnibus (GEO) database. Immunohistochemistry was used to evaluate the expression levels of PTX3, CD68, and CD86 in samples from the human and mouse hepatic IRI models. The effects of PTX3 knockdown or overexpression on macrophage polarization were assessed in Raw264.7. PTX3 knockdown/ overexpression in Raw264.7 and co-culturing with AML12 were performed under conditions of hypoxia-reoxygenation (H/R) to examine pyroptosis and injury in AML12. RESULTS:PTX3 expression was significantly upregulated in both human and mouse hepatic IRI model samples. Bulk and single-cell RNA-seq data analyses revealed that PTX3 is associated with inflammatory response pathways and macrophage activation. Macrophages with high PTX3 expression exhibit M1-like characteristics. Similarly, overexpression of PTX3 promotes M1 polarization of Raw264.7 after H/R, while the knockdown group exhibits reduced M1 polarization. Co-culture results indicated that pyroptosis in AML12 was significantly reduced after H/R in the PTX3 knockdown group, whereas the PTX3 overexpression group exhibited the opposite outcome. CONCLUSION:PTX3 regulates macrophage polarization during hepatic IRI, consequently influencing hepatocellular pyroptosis.

Hepatocellular carcinoma (HCC), one of the most prevalent cancers globally, is closely associated with tumor-associated macrophages (TAMs), including monocyte-derived macrophages and liver-resident Kupffer cells. Understanding TAM heterogeneity at the cellular level is crucial for developing effective HCC prevention and treatment strategies. In this study, we conducted an integrated single-cell analysis of four cohorts (GSE140228, GSE125449, GSE149614 and GSE156625) to elucidate the TAM landscape in HCC. We identified 284 gene markers, termed Panmyeloid markers, that characterize myeloid cells within this context. Our analysis distinguished six clusters of monocyte-derived macrophages (Macro1-Macro6) and four clusters of Kupffer cells (Kupffer1-Kupffer4). Notably, CXCL10 + macrophages and MT1G + Kupffer cells, predominantly located within tumor tissues, exhibited distinct functional characteristics relevant to HCC. We also explored cellular communication between TAMs and T cells, uncovering potential signaling pathways such as the CXCL10/CXCL11-CXCR3 and CXCL12-CXCR4 networks. These findings enhance our understanding of TAMs in HCC and open new avenues for targeted therapeutic interventions.

Rationale: Understanding the immune mechanisms associated with liver transplantation (LT), particularly the involvement of tissue-resident memory T cells (TRMs), represents a significant challenge. Methods: This study employs a multi-omics approach to analyse liver transplant samples from both human (n = 17) and mouse (n = 16), utilizing single-cell RNA sequencing, bulk RNA sequencing, and immunological techniques. Results: Our findings reveal a comprehensive T cell-centric landscape in LT across human and mouse species, involving 235,116 cells. Notably, we found a substantial increase in CD8+ TRMs within rejected grafts compared to stable ones. The elevated presence of CD8+ TRMs is characterised by a distinct expression profile, featuring upregulation of tissue-residency markers (CD69, CXCR6, CD49A and CD103+/-,), immune checkpoints (PD1, CTLA4, and TIGIT), cytotoxic markers (GZMB and IFNG) and proliferative markers (PCNA and TOP2A) during rejection. Furthermore, there is a high expression of transcription factors such as EOMES and RUNX3. Functional assays and analyses of cellular communication underscore the active role of CD8+ TRMs in interacting with other tissue-resident cells, particularly Kupffer cells, especially during rejection episodes. Conclusions: These insights into the distinctive activation and interaction patterns of CD8+ TRMs suggest their potential utility as biomarkers for graft rejection, paving the way for novel therapeutic strategies aimed at enhancing graft tolerance and improving overall transplant outcomes.

Hepatic ischemia-reperfusion (IR) injury significantly impacts liver transplantation success, yet current treatments remain inadequate. This study explores the role of Proto-oncogene serine/threonine-protein kinase (Pim-1) in liver IR, an area previously unexplored. Utilizing a mouse liver IR in vivo model and a MIHA cell hypoxia-reoxygenation in vitro model, we observed that Pim-1 expression increases following IR, inversely correlating with serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. Increased Pim-1 expression stabilizes mitochondrial membranes by modifying Drp1 phosphorylation, reducing mitochondrial fission and apoptosis, thereby mitigating liver damage. Additionally, we discovered that elevated Pim-1 expression is dependent on the trimethylation of histone H3 lysine 9 during liver IR. These findings underscore the importance and potential clinical application of targeting Pim-1 in treating hepatic IR, presenting a novel therapeutic avenue.

Hepatic ischemia‒reperfusion injury is a common injury in liver surgery and liver transplantation that can lead to liver function damage, including oxidative stress, apoptosis, autophagy and inflammatory reactions. Pyroptosis is a type of inflammatory programmed cell death that has been implicated in ischemia‒reperfusion injury-associated inflammatory reactions. Although circular RNAs can regulate cell death in hepatic ischemia‒reperfusion injury, their relationship with pyroptosis remains unclear. Therefore, this study aimed to investigate the effect of circular RNA on pyroptosis in hepatic ischemia‒reperfusion injury. We constructed a mouse hepatic ischemia‒reperfusion injury model for circular RNA sequencing and obtained 40 circular RNAs with significant differential expression, of which 39 were upregulated and 1 was downregulated. Subsequently, the endogenous competitive RNA network was constructed using TarBase, miRTarBase, TargetScan, RNAhybrid, and miRanda. Gene Set Enrichment Analysis, Kyoto Encyclopedia of Genes and Genomes and Gene Ontology functional analyses of downstream target genes revealed that circRNA-Phf21a_0002 might affect pyroptosis by regulating the mTOR signaling pathway and Bach1 by sponging let-7b-5p. The overexpression plasmid upregulated the expression of circRNA-Phf21a_0002 in a hypoxia/reoxygenation model, which aggravated pyroptosis in AML12 cells and apoptosis and necrosis of hepatocytes. Next, we investigated the underlying mechanism and found that circRNA-Phf21a_0002 enabled the expression of Bach1 through sponging of let-7b-5p. The aggravation of pyroptosis via overexpression of circRNA-Phf21a_0002 was reversed by let-7b-5p mimics in hypoxia/reoxygenation-subjected AML12 cells. Collectively, our study clarifies that circRNA-Phf21a_0002 aggravates the pyroptosis of hepatocytes related to ischemia-reperfusion by sponging let-7b-5p. These findings provide new molecular mechanisms and novel biomarkers for follow-up treatment.

Rejection is still a critical barrier to the long-term survival of graft after liver transplantation, requiring clinicians to unveil the underlying mechanism of liver transplant rejection. The cellular diversity and the interplay between immune cells in the liver graft microenvironment remain unclear. Herein, we performed single-cell RNA sequencing analysis to delineate the landscape of immune cells heterogeneity in liver transplantation. T cells, NK cells, B cells, and myeloid cell subsets in human liver and blood were enriched to characterize their tissue distribution, gene expression, and functional modules. The proportion of CCR6+CD4+ T cells increased within an allograft, suggesting that there are more memory CD4+ T cells after transplantation, in parallel with exhausted CTLA4+CD8+ T and actively proliferating MKI67+CD8+ T cells increased significantly, where they manifested heterogeneity, distinct function, and homeostatic proliferation. Remarkably, the changes of CD1c+ DC, CADM+ DC, MDSC, and FOLR3+ Kupffer cells increase significantly, but the proportion of CD163+ Kupffer, APOE+ Kupffer, and GZMA+ Kupffer decreased. Furthermore, we identified LDLR as a novel marker of activated MDSC to prevent liver transplant rejection. Intriguingly, a subset of CD4+CD8+FOXP3+ T cells included in CTLA4+CD8+ T cells was first detected in human liver transplantation. Furthermore, intercellular communication and gene regulatory analysis implicated the LDLR+ MDSC and CTLA4+CD8+ T cells interact through TIGIT-NECTIN2 signaling pathway. Taken together, these findings have gained novel mechanistic insights for understanding the immune landscape in liver transplantation, and it outlines the characteristics of immune cells and provides potential therapeutic targets in liver transplant rejection.

Cuproptosis represents a novel copper-dependent regulated cell death, distinct from other known cell death processes. In this report, a comprehensive analysis of cuproptosis in hepatocellular carcinoma (HCC) was conducted using multi-omics including genomics, bulk RNA-seq, single cell RNA-seq and proteomics. ATP7A, PDHA1 and DLST comprised the top 3 mutation genes in The Cancer Genome Atlas (TCGA)-LIHC; 9 cuproptosis-related genes showed significant, independent prognostic values. Cuproptosis-related hepatocytes were identified and their function were evaluated in single cell assays. Based on cuproptosis-related gene expressions, two immune patterns were found, with the cuproptosis-C1 subtype identified as a cytotoxic immune pattern, while the cuproptosis-C2 subtype was identified as a regulatory immune pattern. Cuproptosis-C2 was associated with a number of pathways involving tumorigenesis. A prognosis model based on differentially expressed genes (DEGs) of cuproptosis patterns was constructed and validated. We established a cuproptosis index (CPI) and further performed an analysis of its clinical relevance. High CPI values were associated with increased levels of alpha-fetoprotein (AFP) and advanced tumor stages. Taken together, this comprehensive analysis provides important, new insights into cuproptosis mechanisms associated with human HCC.

Rejection is still a critical barrier to the long-term survival of graft after liver transplantation, requiring clinicians to unveil the underlying mechanism of liver transplant rejection. The cellular diversity and the interplay between immune cells in the liver graft microenvironment remain unclear. Herein, we performed single-cell RNA sequencing analysis to delineate the landscape of immune cells heterogeneity in liver transplantation. T cells, NK cells, B cells, and myeloid cell subsets in human liver and blood were enriched to characterize their tissue distribution, gene expression, and functional modules. The proportion of CCR6+CD4+ T cells increased within an allograft, suggesting that there are more memory CD4+ T cells after transplantation, in parallel with exhausted CTLA4+CD8+ T and actively proliferating MKI67+CD8+ T cells increased significantly, where they manifested heterogeneity, distinct function, and homeostatic proliferation. Remarkably, the changes of CD1c+ DC, CADM+ DC, MDSC, and FOLR3+ Kupffer cells increase significantly, but the proportion of CD163+ Kupffer, APOE+ Kupffer, and GZMA+ Kupffer decreased. Furthermore, we identified LDLR as a novel marker of activated MDSC to prevent liver transplant rejection. Intriguingly, a subset of CD4+CD8+FOXP3+ T cells included in CTLA4+CD8+ T cells was first detected in human liver transplantation. Furthermore, intercellular communication and gene regulatory analysis implicated the LDLR+ MDSC and CTLA4+CD8+ T cells interact through TIGIT-NECTIN2 signaling pathway. Taken together, these findings have gained novel mechanistic insights for understanding the immune landscape in liver transplantation, and it outlines the characteristics of immune cells and provides potential therapeutic targets in liver transplant rejection.

Bone marrow mesenchymal stem cells (BM-MSCs) regulate the balance between regulatory T cells (Tregs) and T helper 17 (Th17) cells. However, the role of different factors on BM-MSCs-mediated regulation of the Treg/Th17 balance is unknown. BM-MSCs and CD4(+) T lymphocytes were co-cultured with various treatments. The ratio of Treg/Th17 cells was calculated and the expression of different cytokines was measured. BM-MSCs were found to have a proliferative effect on Th17 cells at a basal concentration and at a 2-fold increase in the number of BM-MSCs. However, when the number of BM-MSCs used was increased 4-fold, they had an inhibitory effect on the Th17 cells. The effect of BM-MSCs on Tregs was inhibited by the addition of tacrolimus but not rapamycin. The effect of BM-MSCs on Th17 cells was inhibited by rapamycin. Additionally, the effect of BM-MSCs on Tregs were inhibited by the addition of a transforming growth factor-beta (TGF-beta) blocker, whereas these TGF-beta-blockers had no effect on Th17 cells. Addition of an interleukin (IL)-2 blocker reduced the proportion of Th17 cells when co-cultured with a high number of MSCs compared with the low concentration group and the proportion of Treg cells was significantly decreased when cells were treated with an IL-2 blocker compared with the control group. Together, these results showed the varying effects of MSCs on the ratio of Treg/Th17, its dependence on the number of MSCs and the effects of cytokines in inducing these changes in the balance.

Background: Marrow mesenchymal stem cells (MSCs), especially bone MSCs (BMSCs), play a vital role in immunomodulation of graft rejection but many shortcomings limit its application. In this study, we aimed to investigate the modulation effect of MSCs from gingiva (GMSCs) on the rejection of rat liver transplantation model and to explore the potential mechanisms. Methods: GMSCs were obtained from human gingival tissues and BMSCs were harvested from Brown Norway (BN) rats. The siRNA that mediated knockdown of Fas ligand (FASL) expression in GMSCs (FASL-/-GMSCs) was transfected using lentivirus plasmid. Rat orthotopic liver transplantation model was established. The rats were divided randomly into four groups: normal saline (NS) group, FASL-/-GMSCs group, BMSCs group, and GMSCs group, all of which were injected the solution via dorsal vein of penis to construct the rat rejection model Graft survival and liver function indexes were measured. The immunological reactions of recipients including immune-cytokines, T-helper type 17 (Th17) and regulatory T cells (Treg) were also evaluated by flow cytometry. Results: The graft survival of recipient rats in the GMSCs group was significantly prolonged in comparison with that of the NS, FASL-/-GMSCs and BMSCs group. GMSCs remarkably decreased the levels of AST, ALT and TBIL. As for immune-cytokines, serum levels of IL-2, IFN-c and Th 17 in recipient rats from the GMSCs group reduced significantly compared with that of other three groups, while increased serum IL-10 and TGF-b expressions as well as peripheral serum Treg were also observed in GMSCs group. Conclusion: GMSCs exert immunoprotection on liver transplants by regulating FAS-FASL pathway. The current study firstly provides basis for GMSCs to be used in clinical anti-rejection after liver transplantation. (C) 2020 Published by Elsevier B.V. on behalf of King Saud University.

Objective To explore the expressions of regulatory T cell (Treg) and T helper cell 17 (Th17) during perioperative period of liver transplant (LT) in children of biliary atresia (BA).Methods Peripheral blood samples were harvested from children of surgical observation group at 1 week pre-LT,2 weeks and 4 weeks post-LT respectively.And recipients were followed up for ＞1 year post-LT and healthy controls were designated.The proportions of Treg and Th17 cells in peripheral blood pre-and post-LT were analyzed by flow cytometry.Results The proportions of CD4+ T lymphocytes at different time-points preand post-LT were higher in BA patients than those in healthy control group (P ＜ 0.05).The Treg proportion was higher during pre-LT period (4.04％ ±0.71％) than that at 2 weeks (4.04％ ±0.71％) and 1 month (2.16％ ±0.72％).No difference existed between 2 weeks and 1 month (P＞0.05).The Treg proportions were lower in pediatric receipients than those in health control group (5.06％ ± 1.33％,P＜ 0.05).And the Th17 proportions were higher at 2 weeks (1.43％ ± 0.45％) and 1 month (1.17％ ±0.47％) than those in health control group (0.76％ ± 0.30％,P＜0.05).However,no difference in Th17 proportion existed between pre-LT (1.09％ ± 0.46％) and health control group (P＞0.05).The Treg proportion in recipients at ＞1 year post-LT was higher than that during early post-LT period (P＜0.01).However,no inter-group difference existed in Th17 proportion (P＞0.05).Conclusions The proportion of Treg cells is lower at pre-LT in pediatric BA patients and there is a declining trend at 1 month post-LT.However,the proportion of Th17 cells is higher at pre-LT and it is rising during this period.This change is probably correlated with an immune imbalance and inflammatory response stimulated by a new graft post-LT.

Objective To summarize the efficacy of catheter directed thrombolysis upon postoperative portal vein thrombosis after pediatric living donor liver transplantation .Methods Retrospective analysis was performed for clinical diagnosis and treatment of postoperative portal vein thrombosis after pediatric living donor liver transplantation from January to December 2016 at TJFCH . And the relevant domestic and foreign literature was reviewed . At 2 months after transplantation ,portal vein thrombosis was detected by ultrasonic monitoring at the donor side of portal vein anastomosis . Without delay , transdermal portal vein catheterization was performed . Intraoperatively heparin sodium was dripped into catheter and postoperatively continuous heparin plus urokinase alternately pumped into port vein until a disappearance of thrombus .At the same time ,daily ultrasound was employed for monitoring the change of thrombosis and daily blood test for monitoring coagulation function changes to maintain the level of international normalized ratio (INR) around 2 .0 . After a disappearance of thrombus ,long-term oral warfarin began and continued at least for 1 year . Coagulation function and portal vein blood flow were constantly monitored .Results During the course of treatment ,daily ultrasound showed portal vein thrombosis gradually disappeared .At Day 12 postoperatively ,repeated portography also confirmed a complete disappearance of thrombosis . Regular ultrasonic follow-ups indicated that portal venous blood flow was normal .Conclusions If detected early ,intravenous thrombolysis may be achieved by portal vein catheterization in children after living donor liver transplantation .

Objective To analyze the clinical efficacy and prognosis of living donor liver transplantation (LDLT) in children with biliary atresia (BA).Methods The clinical data of 306 cases of BA patients who received LDLT from June 2013 to December 2017 in the Department of Pediatric Liver Transplantation of Tianjin First Center Hospital were retrospectively analyzed.The incidence of post-LDLT complications was summarized and different factors influencing long-term survival of the recipients were analyzed.Results The median age of recipients at transplantation was 7 (6,9) months,and 88.9％ of the recipients received left lateral lobes.The surgical-related complications mainly included lymphatic leakage (30.7％),bile duct stricture (7.8％) and portal vein stenosis (6.9％).The non-surgical-related complications were mainly EBV infection (57.8％) and CMV infection (36.6％).The incidence of pulmonary infection and acute rejection was 18.6％ and 13.7％,respectively.The 1-,3-,and 5-year survival rates of recipients and grafts were 97.2％,97.2％,97.2％ and 97.2％,96.4％,and 94.6％,respectively.A total number of 8 patients died after LDLT,mainly due to the complications of cardio-pulmonary system.Two patients underwent retransplantation due to graft dysfunction caused by antibody-mediated rejection.Recipient age,PELD scores,GRWR,previous surgical history and matching of ABO blood group between donors and recipients did not affect the long-term survival rates of recipients (P＞0.05).Conclusions Children with biliary atresia who received LDLT can obtain satisfactory clinical results.

OBJECTIVE:To explore the effects of bone marrow mesenchymal stem cells (BMSCs) in different concentrations on the balance of regulatory T cell/T-helper cell 17 (Treg/Th17). METHODS:BMSCs were isolated from SPF grade male Wistar rats with age of 3 weeks old and weight of 50 g. BMSCs were cultured and identified when they were expanded to the 4th generation. CD4+ T lymphocytes were isolated from SPF grade male Wistar rat with age of 6 weeks old and weight of 200 g and assayed for cell purity by flow cytometry. BMSCs were divided into 0.5-fold concentration group, basal concentration group, 2-fold concentration group and 4-fold concentration group by their concentrations of 1×105/well, 2×105/well, 4×105/well and 8×105/well, which were cultured with CD4+ T lymphocytes for 72 hours, respectively. Then the proportion of Treg cells and Th17 cells in each group was detected by flow cytometry, and cytokines were detected by cytometric bead array. RESULTS:The purities of BMSCs and CD4+ T lymphocytes were both higher than 95%. In the co-culture of BMSCs and CD4+ T lymphocytes, the proportions of Treg cells were statistically different among different concentration groups of BMSCs (F = 10.071, P = 0.001), in which BMSCs in 2-fold concentration group had the strongest ability to promote the Treg cells proliferation. The proportion of Treg cells in 2-fold concentration group was significantly higher than that in 0.5-fold concentration group, basal concentration group and 4-fold concentration group [(9.24±2.68)% vs. (3.87±0.38)%, (5.16±1.69)%, (3.86±0.36)%, all P < 0.01]. The level of interleukin-10 (IL-10) was lowest in 0.5-fold concentration group, and it was significantly lower than that in basal concentration group, 2-fold concentration group and 4-fold concentration group (ng/L: 39.80±14.48 vs. 148.43±64.49, 156.40±59.27, 126.92±42.95, all P < 0.05). Transforming growth factor-β (TGF-β) was the highest in basal concentration group, and it was significantly higher than that in 0.5-fold concentration group, 2-fold concentration group and 4-fold concentration group [ng/L: 3.17 (1.88, 5.74) vs. 0.71 (0.32, 1.38), 1.22 (0.47, 2.97), 0.52 (0.37, 1.23), all P < 0.05]. The proportions of Th17 cells were statistically different among the different concentration groups (F = 21.069, P = 0.000), with the highest proportion in basal concentration group which was significantly higher than that in 0.5-fold concentration group or 4-fold concentration group [(0.89±0.08)% vs. (0.64±0.15)%, (0.37±0.10)%, both P < 0.01], but no significant difference was found as compared with 2-fold concentration group [(0.83±0.06)%, P > 0.05]. However, the expressions of IL-17 and IL-6 were not different among the different concentration groups respectively (IL-17: χ2 = 0.550, P = 0.760; IL-6: χ2 = 0.010, P = 0.995). CONCLUSIONS:BMSCs in moderate concentrations [(2-4)×105/well] could promote proliferation both in Treg cells and Th17 cells, but no change could be found in higher concentrations of BMSCs (8×105/well). However, the changes in related cytokines were not synchronized with Treg/Th17 cells.

Objective Hepatic artery thrombosis(HAT) is one of serious complications after liver transplantation. This study was intended to explore the effect of HAT on survival rate of patients/grafts and biliary complications early after liver transplantation with controlled donation after circulatory death (CDCD) donors in children. Methods The clinical data of 236 children with CDCD donor liver transplantation (donor age <14 years) were retrospectively analyzed. Among them, 37 patients developed HAT early postoperatively. The survival rate of patients/grafts and the occurrence of biliary complications were compared between two groups. Results The median follow-up period was 23 months. For HAT (n=37) and non-HAT (n=199) group, the median follow-up period was 27 and 22 months respectively. The 1-year and 3-year survival rates of grafts were 88.2%, 88.2% and 93.2%, 92.4% respectively. And no inter-group statistical difference existed (P=0.373). The 1-year and 3-year graft survival rates were 73.9%, 73.9% and 91.8%, 90.5% respectively. And inter-group statistical difference existed (P<0.01). The incidence of biliary leakage and biliary stricture were 13.5% and 2% and 29.7% and 5.5% respectively. Inter-group statistical differences existed. In HAT group, there were liver failure (n=7, 18.9%) and death (n=3, 8.1%) after transplantation. Conclusions HAT is one of the serious complications after liver transplantation. An early onset of HAT increases the incidence of biliary complications and graft loss in children after CDCD donor liver transplantation. For patients with graft failure, liver re-transplantation may be performed at the right time. Key words: Children; Donor; Liver transplantation; Hepatic artery thrombosis; Biliary complication; Survival rate