Publications by authors named "Lianbao Kong"

23 Publications

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PSMC2 Regulates Cell Cycle Progression Through the p21/Cyclin D1 Pathway and Predicts a Poor Prognosis in Human Hepatocellular Carcinoma.

Front Oncol 2021 26;11:607021. Epub 2021 Feb 26.

Hepatobiliary Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

Proteasome 26S subunit ATPase 2 (PSMC2) plays a pathogenic role in various cancers. However, its function and molecular mechanism in hepatocellular carcinoma (HCC) remain unknown. In this study, tissue microarray (TMA) analysis showed that PSMC2 is highly expressed in HCC tumors and correlates with poor overall and disease-free survival in HCC patients. Multivariate Cox regression analysis revealed that PSMC2 is an independent prognostic factor for HCC patients. Furthermore, our results showed that PSMC2 knockdown inhibited cell proliferation and suppressed tumorigenesis . Knockdown of PSMC2 increased the expression of p21 and therefore decreased the expression of cyclin D1. Dual-luciferase reporter assays indicated that depletion of PSMC2 significantly enhanced the promoter activity of p21. Importantly, PSMC2 knockdown-induced phenotypes were also rescued by downregulation of P21. Taken together, our data suggest that PSMC2 promotes HCC cell proliferation and cell cycle progression through the p21/cyclin D1 signaling pathway and could be a promising diagnostic and therapeutic target for HCC patients.
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http://dx.doi.org/10.3389/fonc.2021.607021DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7952995PMC
February 2021

Macrophage Regnase-1 Deletion Deteriorates Liver Ischemia/Reperfusion Injury Through Regulation of Macrophage Polarization.

Front Physiol 2020 29;11:582347. Epub 2020 Oct 29.

Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

Background: Regnase-1 (MCPIP) has been identified as an anti-inflammatory agent, but little is known about its influence on liver ischemia/reperfusion (I/R) injury. Macrophages can evolve biphasic responses and differentiate into remarkable polarizations, contributing greatly to the uncontrolled inflammatory cascades during liver I/R injury. Therefore, the aim of this study was to explore whether regnase-1 participated in liver I/R via manipulating macrophage polarization.

Materials And Methods: C57BL/6 mice were randomly divided into five groups: Sham, I/R, Clodronate, Clo + BMDM, and Clo + LV MCPIP BMDM. A liver I/R model was established, and histopathological and immunostaining examinations were performed for the liver specimens; double immunofluorescence staining was used to localize MCPIP in the liver. Primary hepatocytes were isolated to simulate a hypoxia and reoxygenation (H/R) model . Bone marrow-derived macrophages (BMDM) were extracted and subjected to lentiviral transduction to knockdown MCPIP expression. BMDM with or without MCPIP deletion were exposed to H/R supernatants, and the polarized states were measured by flow cytometry. RT-PCR analysis and Western blot were also conducted.

Results: Compared to those in the Sham group, liver functions and Suzuki's scores were deteriorated in the I/R group, which were reversed in the Clodronate group. The increased expression of regnase-1 in the I/R group diminished with pretreatment of clodronate liposomes. Subsequent double immunofluorescence staining established the localization of regnase-1 in macrophages in the liver. The insulted lesions in the Clodronate group became progressively aggravated with adoptive transfer of BMDM in the Clo + BMDM group, and they were further exacerbated with the transfusion of BMDM with MCPIP knockdown in the Clo + LV MCPIP BMDM group. Gene expressions of M1 and M2 markers were detected by RT-PCR, suggesting that MCPIP knockdown tended to favor the M1 transformation. Subsequently, flow cytometrical detection showed that, upon stimulation by H/R supernatants, LV-MCPIP BMDM posed a higher ratio of M1/M2 than BMDM. Finally, we found that MCPIP participated in macrophage M1/M2 polarization through the NF-κB, C/EBPβ, and PPARγ signaling pathways during liver I/R.

Conclusion: Our study confirms that regnase-1 plays a critical role in liver I/R via regulation of macrophage polarization and, thus, might offer a potential therapeutic target.
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http://dx.doi.org/10.3389/fphys.2020.582347DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7658104PMC
October 2020

N-acetylcysteine alleviates liver injury by suppressing macrophage-mediated inflammatory response post microwave ablation.

Int Immunopharmacol 2020 Aug 18;85:106580. Epub 2020 May 18.

Hepatobiliary Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China. Electronic address:

Object: To investigate N-acetyl-cysteine (NAC) would able to alleviate liver injury and systemic inflammatory response caused by microwave ablation (MWA) in rats.

Materials And Methods: Male Sprague-Dawley rats weighing 150-200 g were randomly divided into sham group (only anesthesia and laparotomy except MWA but with intraperitoneal PBS or NAC solution injection according to different situations), control group (intraperitoneal PBS injection for comparation 2 h prior to MWA), and NAC-treated group (intraperitoneal N-acetyl-cysteine (300 mg/kg) injection 2 h prior to MWA). Experimental rats were sacrificed at 4 h following operation in line with the liver injury severity curve. Liver tissue and serum samples were collected for determination of pathology, apoptosis, macrophages contents and protein expression.

Results: The elevated serum level of liver enzymes, Myeloperoxidase (MPO) and inflammatory factors (TNF-α and CXCL1) in MWA-treated rats revealed injurious and pro- inflammatory effect of MVA. Macrophages aggregation was detected in MWA exposure rats similarly. and NAC pre-conditioning mitigate liver damage and hepatocyte apoptosis, besides macrophages accumulation and following inflammatory response in liver tissue.

Conclusion: Our results demonstrated that N-acetyl-cysteine application alleviate macrophages aggregation and inflammatory response in liver suffering microwave ablation, and mitigating liver injury and cell apoptosis.
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http://dx.doi.org/10.1016/j.intimp.2020.106580DOI Listing
August 2020

MiR-3174 promotes proliferation and inhibits apoptosis by targeting FOXO1 in hepatocellular carcinoma.

Biochem Biophys Res Commun 2020 06 10;526(4):889-897. Epub 2020 Apr 10.

Hepatobiliary Center, The First Affiliated Hospital of Nanjing Medical University, Key Laboratory of Liver Transplantation, Chinese Academy of Medical Sciences, NHC Key Laboratory of Living Donor Liver Transplantation (Nanjing Medical University), Nanjing, Jiangsu Province, China. Electronic address:

Introduction: MicroRNAs (miRNAs) have been confirmed to play a crucial part in oncogenesis. Several studies suggested that MiR-3174 act as a tumor promoter in various Malignant neoplasm. However, the biological function of miR-3174 in hepatocellular carcinoma (HCC) still highly unexplored.

Methods: We screened differentially over-expressed miRNAs by The Cancer Genome Atlas (TCGA) and the GEO databases. The expression of miR-3174 in HCC cells and tissues was detected by qRT-PCR. The cellular behaviors of transfected cells were respectively examined by colony formation assays, EdU Assays and flow cytometry. Forkhead box O1 transcription factor (FOXO1) was predicted and confirmed as a direct target of miR-3174 by bioinformatics analysis and dual-luciferase reporter gene assay.

Results: MiR-3174 was up-regulated in HCC tissues and cells, and the expression level of it was highly associated with tumor size and Edmondson grade. Our study pioneering validates that upregulated miR-3174 promote cell proliferation and inhibit cell apoptosis in vitro and in vivo. Meanwhile, our study verified that miR-3174 regulate Bim, P21, cyclin D1 and c-MYC expression by directly targeting FOXO1.

Conclusion: The upregulated miR-3174 promote cell proliferation and inhibit cell apoptosis by downregulating FOXO1 expression in HCC. MiR-3174 may be a novel candidate for targeted delivery of miRNA therapeutics for HCC patients.
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http://dx.doi.org/10.1016/j.bbrc.2020.03.152DOI Listing
June 2020

Roquin-1 Regulates Macrophage Immune Response and Participates in Hepatic Ischemia-Reperfusion Injury.

J Immunol 2020 03 29;204(5):1322-1333. Epub 2020 Jan 29.

Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, 210029 Nanjing, People's Republic of China

With the development of liver surgery, ischemia-reperfusion (IR) injury has received increasing attention. Roquin-1 has been shown to play an important role in innate immune and immune balance. We demonstrate that Roquin-1 expression increased at 1 h after IR and then decreased in C57B/L mice. The immunofluorescence double-label showed that Roquin-1 was mainly expressed in macrophages (mø). Furthermore, we used clodronate liposomes to remove mø, and injected the bone marrow-derived mø (BMDM) through the tail vein in 1 h before IR. We found that liver IR injury was aggravated by Roquin-1 interference. The results of PCR and ELISA suggested that after interference with Roquin-1, mø increased toward M1 and decreased toward M2. Then, interference with Roquin-1 promoted the polarization of mø to M1 and inhibited the polarization of M2. By Western blot technology and AMPKα and mTOR inhibitors, we found that Roquin-1 promotes the phosphorylation of mTOR and STAT3 by inhibiting the phosphorylation of AMPKα. We used AICAR to activate AMPKα in mø and found that the level of ubiquitination of AMPKα was decreased after activation of AMPKα. Furthermore, by bioinformatics methods, we identified potential ubiquitination sites on AMPKα. By the point mutation experiments in vitro, we confirmed that the ubiquitination of these sites is regulated by Roquin-1. Meanwhile, Roquin-1 interference inhibited the activation and function of AMPKα. This topic describes the protection of liver IR injury by Roquin-1 and discusses its main mechanism for regulating AMPKα activity through ubiquitination and affecting the polarization of mø.
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http://dx.doi.org/10.4049/jimmunol.1900053DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7026771PMC
March 2020

Precoagulation with microwave ablation for hepatic parenchymal transection during liver partial resection.

Int J Hyperthermia 2019 28;36(1):146-150. Epub 2018 Nov 28.

b Hepatobiliary/Liver Transplantation Center , The First Affiliated Hospital of Nanjing Medical University , Nanjing , China.

Purpose: To evaluate the feasibility of precoagulation with microwave ablation (MWA) for hepatic parenchymal transection during liver partial resection.

Methods: A total of 66 eligible patients were enrolled in this double-blind, randomized, controlled study. Patients were randomized to receive either the traditional clamp-crushing method (Control group) or the MWA precoagulation method (MWA group) for hepatic parenchymal transection during liver partial resection. The operative time, hepatic portal occlusion time, intraoperative blood loss and transfusion, postoperative complications and recovery outcomes were compared.

Results: Compared to the Control group, the MWA group had significantly less intraoperative blood loss. Fewer red blood cell transfusions were observed in the MWA group but without statistical significance. The MWA group showed significantly higher serum alanine aminotransferase and aspartate aminotransferase levels at day 1 postoperatively, but no differences between the MWA and Control groups were found at days 3 and 7. There were no significant differences in terms of operative time, hepatic portal occlusion time, postoperative total bilirubin levels, human albumin solution consumption or length of hospital stay. Postoperative complications such as impaired renal function, pyrexia, admission to ICU, abscess, biliary leakage, intrahepatic and distant tumor recurrence and in-hospital mortality were comparable between the two groups.

Conclusion: Precoagulation with MWA reduced intraoperative blood loss with similar postoperative complications, providing a safe, effective, novel alternative for hepatic parenchymal transection during liver partial resection. Additional results from larger series are recommended to confirm these findings.
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http://dx.doi.org/10.1080/02656736.2018.1540799DOI Listing
January 2020

Exosomes Derived from Dendritic Cells Attenuate Liver Injury by Modulating the Balance of Treg and Th17 Cells After Ischemia Reperfusion.

Cell Physiol Biochem 2018 29;46(2):740-756. Epub 2018 Mar 29.

Background/aims: The present study aimed to evaluate the effects as well as the underlying mechanisms of bone marrow-derived dendritic cells (BMDCs) and exosomes produced by BMDCs (DEXs) on hepatic ischemia-reperfusion (I/R) injury (IRI).

Methods: Primary hepatocytes were isolated and used to mimic the liver IR microenvironment. BMDCs were induced and characterized both biochemically with a flow cytometer (FCM) and biophysically with a microscope. Then, we exposed BMDCs to the supernatants from primary hepatocytes and evaluated the maturation of BMDCs by FCM. BMDCs were systemically injected into mice before liver IR via the tail vein, and the therapeutic effects were evaluated. The serum levels of transaminases (aspartate aminotransferase (AST) and alanine aminotransferase (ALT), inflammatory cytokines, and histological changes were respectively examined by ELISA, RT-qPCR and microscopy. Furthermore, we isolated DEXs by ultracentrifugation, characterized DEXs by transmission electron microscopy (TEM) and nanosight tracking analysis (NTA) and western blotting (WB), and then we co-cultured BMDCs/DEXs and naïve T cells and performed FCM, ELISA and confocal imaging. Moreover, we injected DEXs into mice prior to liver IR via the tail vein and examined its therapeutic effects by microscopy and ELISA. Finally, inhibitors of HSP70 (cmHSP70.1), PI3K (BKM120) and mTOR (Rapamycin) were used to investigate the role of HSP70 and the PI3K/mTOR axis in the effects of DEXs on naïve T cells by WB and FCM.

Results: Bone marrow cells were efficiently induced into dendritic cells (DCs) with typical DC characteristics. The supernatants from primary hepatocytes exposed to H/R upregulated DC maturation markers. After DC administration, liver IR injury was improved with histopathological scores and serum transaminases. Additionally, we found that the anti-inflammatory cytokines TGF-β, Foxp3 and interleukin (IL)-10 were upregulated and that IL-17 was downregulated. Furthermore, confocal imaging revealed that the uptake of H/R-DEXs by naïve T cells was greater than that of DEXs derived from the control or negative group of BMDCs, and this increase was correlated with a significantly greater degree of differentiation of Tregs and Th17 cells. Moreover, H/R-DEXs administration improved liver function in mice after IR. Finally, the inhibition of HSP70, PI3K and mTOR completely abolished the effect of DEXs on naïve T cells.

Conclusion: These results demonstrated that BMDCs and DEXs could alleviate hepatic I/R injury via modulating the balance between Tregs and Th17 cells. DEXs transported HSP70 into naïve T cells and stimulated the PI3K/mTOR axis to modulate the balance between Tregs and Th17 cells and protect the liver from IR injury.
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http://dx.doi.org/10.1159/000488733DOI Listing
July 2018

Mir-24 regulates hepatocyte apoptosis via BIM during acute liver failure.

Am J Transl Res 2017 15;9(11):4925-4935. Epub 2017 Nov 15.

Department of Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical UniversityNo. 300, Guangzhou Road, 210029 Nanjing, China.

Acuteliver failure (ALF) has a high mortality rate and is characterized by massive hepatocyte destruction. Although microRNAs (miRNAs) play an important role in manyliver diseases, the role of miRNAs in ALF development is unknown. In this study, the murine ALF model was induced by intraperitoneal injection of D-galactosamine/lipopolysaccharide (D-GalN/LPS). Compared with saline-treated mice, miR-24 was distinctly down-regulated post D-GalN/LPS challenge and D-galactosamine/tumor necrosis factor (D-GalN/TNF) challenge , which was confirmed by quantitative real-time polymerase chain reaction. Meanwhile, the mRNA and protein levels of the BH3-only-domain-containing protein BIM were upregulated after challenge both and . Previous studies have demonstrated that hepatocyte apoptosis is a distinguishing feature of D-GalN/LPS-associated liver failure. In this study, D-GalN/LPS-challenged mice showed higher alanine aminotransferase and aspartate aminotransferase levels, more severe liver damage, increased numbers of apoptotic hepatocytes and higher levels of caspase-3 compared with saline-treated mice. In D-GalN/TNF-treated BNLCL2 cells, miR-24 overexpression attenuated apoptosis.Furthermore, miR-24 overexpression reduced BIM mRNA and protein levels . Taken together, these findings demonstrate that miR-24 regulates hepatocyte apoptosis via BIM during ALF development, suggesting that miR-24 is a novel onco-miRNA that may provide potential therapeutic targets for ALF.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5714776PMC
November 2017

Application of microwave ablation in the emergent control of intraoperative life-threatening tumor hemorrhage during hepatic surgeries.

Int J Hyperthermia 2018 11 23;34(7):1049-1052. Epub 2017 Oct 23.

a Department of Liver Surgery, The First Affiliated Hospital of Nanjing Medical University, Key Laboratory of Living Donor Liver Transplantation , National Health and Family Planning Commission , Nanjing , China.

Purpose: This study was designed to evaluate the efficacy and safety of microwave ablation (MWA) in the treatment of intraoperative life-threatening tumour haemorrhage during hepatic surgeries.

Methods: Three cases of MWA application in the emergent control of life-threatening hepatic tumour haemorrhage were analysed and reported.

Results: Satisfactory hemostasis for hepatic tumour rupture was achieved by MWA in all three cases. No major complications, such as post-operative haemorrhage, bile duct injury, liver abscess, colon perforation, skin burns, tumour seeding or renal dysfunction, were identified.

Conclusions: MWA may be a feasible, effective and simple strategy for the emergent control of intraoperative hepatic tumour bleeding. To the best of our knowledge, this study represents the first reported cases of this novel application of MWA.
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http://dx.doi.org/10.1080/02656736.2017.1388929DOI Listing
November 2018

Interleukin-11 protects mouse liver from warm ischemia/reperfusion (WI/Rp) injury.

Clin Res Hepatol Gastroenterol 2016 Nov 23;40(5):562-570. Epub 2016 Mar 23.

Department of Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, No. 300, Guangzhou Road, 210029 Nanjing, China. Electronic address:

Background: IL-11 is a multifunctional cytokine that belongs to the IL-6 family. Previous studies have demonstrated that IL-11 has underlying anti-inflammatory and anti-apoptotic properties. In this study, we evaluated the potential effects of IL-11 on mouse liver WI/Rp injury.

Methods: For in vivo experiments, mice were randomly divided into four main experimental groups (n=5 each): (1) normal group - anesthesia; (2) sham group- laparotomy; (3) I/R group- liver WI/Rp; and (4) IL-11 pretreatment (500μg/kg, tail vein injection) group- administration of RhIL-11 2h before liver WI/Rp induced in the same manner as in group 3. For in vitro experiments, cells were divided into two groups: (1) H/R group- H/R; and (2) IL-11 pretreatment group- pretreatment with RhIL-11 (2μg/mL for 12h) before the induction of H/R. For both groups, three periods of reoxygenation were examined (2h, 6h, and 12h).

Results: In the in vivo experiments, IL-11 protected mouse livers from WI/Rp by reducing liver enzyme levels and cellular degeneration. In the in vitro experiments, IL-11 significantly reduced hepatocyte apoptosis. In both the in vivo and in vitro experiments, IL-11 pre-treatment significantly reduced the expression of TNF-α and IL-1β. In addition, NF-κB, a target of IL-11, was suppressed in macrophages after IL-11 pre-treatment.

Conclusions: Pre-treatment with IL-11 protects mouse livers from WI/Rp injury by suppressing NF-kB activity.
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http://dx.doi.org/10.1016/j.clinre.2015.11.009DOI Listing
November 2016

[Impact of microwave dealing with the cutting surface on the hepatocellular carcinoma recurrence after hepatectomy].

Zhonghua Zhong Liu Za Zhi 2015 Dec;37(12):909-12

Liver Transplantation Center, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China; Email:

Objective: To explore the impact of microwave dealing with cutting surface on perioperative liver function recovery and recurrence and metastasis after hepatectomy for HCC.

Methods: Clinical data of 133 patients with HCC from March 2009 to November 2010 were retrospectively analyzed. They were divided into the conventional surgery group (66 cases) and microwave treatment group (67 cases). A domestic ECO-100 microwave knife was inserted into the liver cutting surface 0.5 cm from the cutting edge, and repeated multi-point burning with an average time of 25 minutes in the microwave treatment group. Then the perioperative liver function recovery and recurrence and metastasis in the two groups were compared.

Results: The operation time of conventional surgery group was (158.0 ± 31.0) minutes, and that of microwave treatment group was significantly longer (181.0 ± 28.0) minutes (P=0.027). There were no significant differences in the liver function recovery between the two groups (P>0.05). There were 6 cases of recurrence and metastasis after 6 months and 9 cases after 12 months in the microwave treatment group, while there were 15 cases of recurrence and metastasis after 6 months and 20 cases after 12 months in the conventional surgery group, showing a significant difference (P=0.034 and 0.022, respectively).

Conclusions: Microwave dealing with the cutting surface has no significant effect on perioperative liver function recovery in hepatectomy. However, microwave treatment can reduce the in situ recurrence in HCC patients within the first year after surgery, indicating a good clinical application value.
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December 2015

The microRNA-146a/b attenuates acute small-for-size liver graft injury in rats.

Liver Int 2015 Mar 17;35(3):914-24. Epub 2014 Sep 17.

Department of Neonatal Surgery, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.

Background & Aims: A critical role of the Toll-like receptor (TLR)-4 and its downstream mediators in the pathogenesis of small-for-size liver graft injury has been documented. Recently, the microRNA-146 (miR-146) was identified as a potent negative regulator of the TLR4 signalling pathway. In this study, the role of miR-146a and miR-146b in the attenuation of TLR-4 signalling and small-for-size liver graft injury was investigated.

Methods: The expression levels of miR-146a and miR-146b during small-for-size liver graft injury were studied in vivo. In addition, the effects of miR-146a and miR-146b on the expression of IRAK1 and TRAF6 in the rat macrophage cell line NR8383 and rat liver kupffer cells were studied in vitro. The in vivo effect of miR-146a and miR-146b on small-for-size liver graft injury was studied by the tail vein injection of miR-146a mimics and miR-146b mimics.

Results: The levels of miR-146a and miR-146b decreased with a small-for-size liver graft. MiR-146a and miR-146b inhibited IRAK1 and TRAF6 expression by binding to the 3'UTR of IRAK1 or TRAF6, respectively, in the rat macrophage cell line NR8383. The administration of miR-146a mimics and miR-146b mimics prevented liver graft injury in small-for-size liver graft injury via the inactivation of IRAK1 and TRAF6 in vivo.

Conclusions: miR-146a and miR-146b prevent liver injury in small-for-size liver graft injury via the inactivation of IRAK1 and TRAF6.
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http://dx.doi.org/10.1111/liv.12674DOI Listing
March 2015

miR-146a ameliorates liver ischemia/reperfusion injury by suppressing IRAK1 and TRAF6.

PLoS One 2014 2;9(7):e101530. Epub 2014 Jul 2.

Department of Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.

A critical role of the Toll-like receptor(TLR) and its downstream molecules, including IL-1 receptor-associated kinase 1(IRAK1) and tumor necrosis factor receptor- associated factor 6(TRAF6), in the pathogenesis of liver ischemia/reperfusion (I/R) injury has been documented. Recently a microRNA, miR-146a, was identified as a potent negative regulator of the TLR signaling pathway. In this study, we investigated the role of miR-146a to attenuate TLR signaling and liver I/R injury in vivo and in vitro. miR-146a was decreased in mice Kupffer cells following hepatic I/R, whereas IRAK1 and TRAF6 increased. Overexpression of miR-146a directly decreased IRAK1 and TRAF6 expression and attenuated the release of proinflammatory cytokines through the inactivation of NF-κB P65 in hypoxia/reoxygenation (H/R)-induced macrophages, RAW264.7 cells. Knockdown experiments demonstrated that IRAK1 and TRAF6 are two potential targets for reducing the release of proinflammatory cytokines. Moreover, co-culture assays indicated that miR-146a decreases the apoptosis of hepatocytes after H/R. In vivo administration of Ago-miR-146a, a stable version of miR-146a in vivo, protected against liver injury in mice after I/R via inactivation of the TLR signaling pathway. We conclude that miR-146a ameliorates liver ischemia/reperfusion injury in vivo and hypoxia/reoxygenation injury in vitro by directly suppressing IRAK1 and TRAF6.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0101530PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4079695PMC
October 2015

Comparative proteome profile during the early period of small-for-size liver transplantation in rats revealed the protective role of Prdx5.

J Hepatol 2010 Jul 3;53(1):73-83. Epub 2010 Apr 3.

Key Laboratory of Living Donor Liver Transplantation, Ministry of Public Health, Department of Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.

Background & Aims: In living-donor liver transplantation (LDLT), "small-for-size graft (SFSG) syndrome" is a complex process resulting primarily from ischemia-reperfusion injury (IRI) and portal hypertension associated with size mismatch between graft and recipient. In the early period of LDLT, molecular events related to subsequent apoptosis, necrosis, proliferation and regeneration appeared in specific protein expression patterns.

Methods: We used 2D-PAGE and MALDI-TOF/TOF technology to construct a comparative proteome profile for small-for-size liver grafts (SFSGs) during the early period of LDLT in rats (ischemia 1h, and 2, 6, 24, 48 h post-reperfusion); sham-operated liver was the control. Western blotting was used to confirm the proteomics results and immunohistochemistry was carried out to explore the cellular localization of selected proteins. We further performed cluster and bioinformatics analyses of differential proteins. Lastly, we overexpressed Prdx5 in liver grafts using an adenoviral vector to evaluate its protective role.

Results: We identified 314 differential protein spots corresponding to 259 different proteins. Cluster analyses revealed six expression patterns, and bioinformatics analyses revealed that each pattern was related to many specific cell processes. We also showed that Prdx5 overexpression could attenuate injury to SFSGs and increase survival in recipients.

Conclusions: Taken together, these results reveal an important proteome profile that is functional in SFSGs during early period of LDLT, and provide a strong basis for further research.
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http://dx.doi.org/10.1016/j.jhep.2010.01.032DOI Listing
July 2010

Over-expression of Toll-like receptors and their ligands in small-for-size graft.

Hepatol Res 2010 Apr 11;40(4):318-29. Epub 2010 Jan 11.

Department of Neonatal Surgery, Nanjing Children's Hospital Affiliated to Nanjing Medical University.

Aim: Toll-like receptors (TLRs) participate in several physiological and pathological processes of transplantation, including inflammation and allograft rejection, but the expression of TLRs and their ligands remains undetermined in small-for-size graft transplantation.

Methods: A non-arterialized partial liver transplantation model was used. The expression of TLR2 and TLR4 mRNA and protein, CD14 and Myeloid Differentiation-2 (MD-2) mRNA, as well as TLR2 and TLR4 exogenous ligands (endotoxin) and endogenous ligands [heat shock protein (HSP) 60 and 70] were assessed. The signaling pathways induced by TLR2 and TLR4 were also assessed.

Results: In small-for-size liver graft, the expression of mRNA and protein of TLR2 and TLR4, CD14 and MD-2 mRNA, as well as endogenous ligands of TLR2 and TLR4 such as HSP60 and HSP70 was quickly and significantly increased after reperfusion, and reached a peak at 3 h after reperfusion. The levels of exogenous ligands (endotoxin) were increased and reached a peak at 6 h after reperfusion. The appearance of TLR2 and TLR4 mRNA was accompanied by increased HSP 60 and 70 mRNA within 24 h after reperfusion. In the small-for-size group, the peak levels of TLRs and their endogenous ligands appeared earlier than those in the full size group; the peak levels of TLRs and their endogenous and exogenous ligands were higher than those in the full size group.

Conclusion: TLR2 and TLR4, as well as their endogenous and exogenous ligands were activated in small-for-size liver graft transplantation.
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http://dx.doi.org/10.1111/j.1872-034X.2009.00603.xDOI Listing
April 2010

Antifibrotic effect of hepatocyte growth factor-expressing mesenchymal stem cells in small-for-size liver transplant rats.

Stem Cells Dev 2010 Jun;19(6):903-14

Key Laboratory of Living Donor Liver Transplantation, Ministry of Public Health, The First Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu, People's Republic of China.

Ischemia-reperfusion and chronic injuries associated with small-for-size liver transplantation (SFSLT) impair the regeneration of liver graft and induce liver fibrosis. Mesenchymal stem cells (MSCs) can prevent the development of liver fibrosis, and hepatocyte growth factor (HGF) can also attenuate liver cirrhosis. Our previous studies have demonstrated that higher occurrence of liver fibrosis existed in rats post-SFSLT, and that implantation of HGF/MSCs, the human HGF (hHGF)-expressing MSCs, can improve liver regeneration, reduce mortality of rats, as well as have the potent antifibrotic effect in this SFSLT model. In the present study, we implanted HGF/MSCs into liver grafts via the portal vein and investigated their role in antifibrosis effect, using a 30% SFSLT rat model. Fibrosis indexes, including laminin (LN), hyaluronic acid (HA) levels in serum and hydroxyproline (Hyp) content in the liver grafts, the expression of transforming growth factor-beta1 (TGF-beta(1)), rat HGF (rHGF), alpha-smooth muscle actin (alpha-SMA) in hepatic stellate cells (HSCs), alanine aminotransferase (ALT), total bilirubin (BIL), and albumin (ALB) levels in serum, in rats in different treatment groups were assessed at different time points. We found that HGF/MSCs significantly inhibited the formation of liver fibrosis in rats undergoing SFSLT, while MSCs and HGF had synergistic effects in the process. The antifibrosis effect of HGF/MSCs may have contributed in modulating the activation and apoptosis of HSCs, elevating the rHGF expression level, and decreasing the TGF-beta(1) secretion of activated HSCs. These studies suggest that HGF/MSCs may be a novel therapeutic option for the treatment of liver fibrosis after SFSLT.
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http://dx.doi.org/10.1089/scd.2009.0254DOI Listing
June 2010

Allograft inflammatory factor-1 is up-regulated in warm and cold ischemia-reperfusion injury in rat liver and may be inhibited by FK506.

J Surg Res 2011 Jan 25;165(1):158-64. Epub 2009 Jun 25.

Department of Neonatal Surgery, Nanjing Children's Hospital Affiliated to Nanjing Medical University, Nanjing, China.

Background: Allograft inflammatory factor-1 (AIF-1) plays an important role in immune response and vasculopathy in allografts. The present study investigated activation of AIF-1 in warm and cold ischemia-reperfusion (IR) injury of rat liver, and the potential inhibitory effect of FK506.

Methods: We used warm IR injury, orthotopic transplantation, and allograft rejection models in this study. We assessed expression of AIF-1 mRNA and protein, as well as its inducers interferon-γ (IFN-γ) and interleukin-1β (IL-1β). The potential inhibitory effect of FK506 on AIF-1 in a rat macrophage cell line and in these three models was also assessed.

Results: AIF-1 mRNA and protein, as well as its inducers IFN-γ and IL-1β, were significantly increased in the warm IR injury, orthotopic transplantation, and allograft rejection models. Real-time RT-PCR and Western blotting showed that pretreatment with low-dose FK506 partially inhibited AIF-1 activation as well as its inducers (IFN-γ and IL-1β) in these three models. Western blotting showed that IFN-γ and IL-1β activated AIF-1 in a macrophage cell line, but pretreatment with FK506 did not inhibit AIF-1 activation in vitro. Hematoxylin and eosin staining showed that edema and necrosis in the liver, as well as alanine aminotransferase (ALT) in serum, of these three groups was reduced after FK506 pretreatment.

Conclusion: AIF-1 was activated in warm and cold IR injury, and pretreatment with low-dose FK506 partly inhibited AIF-1 activation and reduced warm and cold IR injury.
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http://dx.doi.org/10.1016/j.jss.2009.05.038DOI Listing
January 2011

The effect of deoxyschisandrin on blood tacrolimus levels: a case report.

Immunopharmacol Immunotoxicol 2010 Mar;32(1):177-8

The First Affiliated Hospital, Department of Liver Transplantation Center, and The National Institute of Living Donor Liver Transplantation, Nanjing Medical University, Nanjing, China.

Background And Aim: It is well known that tacrolimus (FK-506 or Prograf [Tac]) is widely used to prevent allograft rejection after liver transplantation. Herein, we report a case of a living-donor liver transplant recipient who displayed increased Tac level during concomitant usage of deoxyschisandrin.

Materials And Methods: The recipient is a 42-year-old male who received a liver transplant from his sister for hepatitis B-related hepatocirrhosis. Immunosuppression was initiated with Tac, corticosteroids, and mycophenolate mofetil. Fifteen months after transplantation, he received Tac at an oral dose of 6mg per day alone to avoid allograft rejection. Then he experienced over low Tac concentration, diarrhea and an increase in serum alanine aminotransferase (ALT) during this phase. Thus, he was readmitted to the hospital two days later. Because of unstable liver function, deoxyschisandrin therapy was initiated at a dose of 22.5mg after each meal.

Results: Three days later, the blood Tac concentration increased from 2.3ng/mL to 17.7ng/mL.

Conclusions: Deoxyschisandrin could markedly increase the blood level of Tac in this liver transplant patient. Monitoring the trough levels of Tac frequently is recommended when deoxyschisandrin is co-administered.
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http://dx.doi.org/10.1080/08923970903117365DOI Listing
March 2010

Emergency adult living donor right lobe liver transplantation for fulminant hepatic failure.

Front Med China 2007 Jul 1;1(3):282-6. Epub 2007 Jul 1.

Key Laboratory of Living Donor Liver Transplantation of Ministry of Public Health, Liver Transplantation Center, the First Affiliated Hospital of Nanjing Medical University, Nanjing, 210029, China.

Fulminant hepatitis is fatal in most cases and timely liver transplantation is the only effective treatment. This study evaluates the survival outcomes of patients who underwent living-donor liver transplantation (LDLT) using right lobe liver grafts for fulminant liver failure due to hepatitis B infection. Nine cases of adult right lobe LDLT were performed in our department from September 2002 to August 2005 and the clinical and following-up data were reviewed. According to the pre-transplant Child-Pugh-Turcotte classification, the nine patients were classified as grade C. The model for end-stage liver disease (MELD) score of these patients ranged from 16 to 42. The principal complications before transplantation included abnormal renal function, hepatic coma of different degrees and alimentary tract hemorrhage. The main complications after transplantation included pulmonary infection in two cases, acute renal failure in three cases and transplantation-related encephalopathy in one case. No primary failure of vascular or biliary complications occurred. The one-year survival rate was 55.6%. There were no serious complications or deaths in donors. In general, it is extremely difficult to treat fulminant hepatitis by conservative regimen, particularly, in cases with rapid progression. Emergency adult living-donor liver transplantation is an effective treatment for fulminant hepatitis patients and is relatively safe for donors.
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http://dx.doi.org/10.1007/s11684-007-0054-yDOI Listing
July 2007

Impaired hepatic regeneration by ischemic preconditioning in a rat model of small-for-size liver transplantation.

Transpl Immunol 2007 Jul 13;18(1):37-43. Epub 2007 Mar 13.

Liver Transplantation Center, The First Affiliated Hospital of Nanjing Medical University, 300 GuangZhou Road, Nanjing, 210029, China.

Objective: Graft size is one of the major risk factors in adult-to-adult living donor liver transplantation and rapid regeneration is an essential post-operative requirement. Ischemic preconditioning (IPC) has been shown to be an effective strategy in the reduction of hepatic ischemia-reperfusion injury and stimulation of liver regeneration. This study was designed to evaluate the effects of IPC on liver regeneration in small-for-size liver grafts.

Methods: We employed a rat orthotopic liver transplantation model using small-for-size (30%) grafts, in the presence or absence (control) of IPC (10 min of ischemia followed by 15 min of reperfusion). Survival rate, graft injury, hepatocellular proliferation, cell cycle progression, Stat3 activation, as well as TNF-alpha and IL-6 expression were assessed.

Results: IPC significantly enhanced the extent of graft injury and hindered hepatic regeneration in small-for-size liver grafts. The 7-day survival rate was also reduced by IPC, but failed to reach statistical significance. IPC did not affect TNF-alpha levels, but significantly decreased the elevation of IL-6 after reperfusion. These findings were correlated with down-regulation of cyclin E and cyclin D1, and decreased numbers of PCNA-positive nuclei in IPC grafts. These results were inconsistent with Stat3 activation, as P-Stat3 exhibited a stronger and prolonged pattern of expression in the IPC group, compared to controls.

Conclusions: Ischemic preconditioning may impair liver regeneration in small-for-size liver grafts by decreasing IL-6 and blunting cell cycle progression, through a mechanism at least partially independent of Stat3.
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http://dx.doi.org/10.1016/j.trim.2007.02.002DOI Listing
July 2007

[Multimodal approach to clinical liver transplantation].

Zhonghua Wai Ke Za Zhi 2002 Oct;40(10):758-61

Department of Hepatobiliary, First Affiliated Hospital, Nanjing Medical University, Nanjing 210029, China.

Objective: To sum up the clinical experience of liver transplantation.

Method: A retrospective study was made in 11 patients receiving living donor liver transplantation (LDLT)/and 14 patients having orthotopic liver transplantation (OLT), including one time operation of reduced size liver retransplantation and one time operation of cadaveric liver retransplantation.

Results: The voluntary donors were a sister and 10 mothers of recipients. The location of graft included 3 patients of segment II, III, part of IV (not including intermediate hepatic veins), 6 patients of segment II, III, IV (including intermediate hepatic veins), and 2 patients of V, VI, VII, VIII (not including intermediate hepatic veins). The weight range of graft was 270 - 620 g. Twenty-four recipients achieved a long-term survival and retained normal liver function during the follow-up. Only 1 patient died from serious rejection on the 72nd day postoperatively. Ten patients with hepatitis B cirrhosis were treated with lamivudine and anti-HBVIg, and HBV-DNA in serum was negative during the follow-up for 4 approximately 21 months. Copperoxidase, ceruloplasmin and main indexes of liver function became normal in all patients with Wilson's Disease. Postoperative complications included abdominal hemorrhage (2 patients), acute respiratory distress syndrome (5), acute rejection (4), and acute renal function failure (2).

Conclusions: The wise solution to improve the result of liver transplantation and optimize liver resources is the "multimodal approach", by which all kinds of techniques for liver transplantation including CLT, LDLT and RSLT should well developed.
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October 2002

[A clinical report of 12 case-times of living related liver transplantation].

Zhonghua Yi Xue Za Zhi 2002 Apr;82(7):435-9

Department of Liver Surgery, First Affiliated Hospital of NanJing Medical University, Liver Transplantation Center for Jiangsu Province, Nanjing 210029, China.

Objective: To evaluate and summarize the clinical application of living related liver transplantation (LRLT).

Methods: A retrospective analysis was made in altogether 12 LRLT operations of living related liver transplantation performed in our department. The indication and timing, surgical complications, and nonsurgical issues including infection, rejection, advantages of LRLT in our series were reviewed.

Result: All the 11 donors are uneventfully after operation; the first receptor was dead, and 10 receptors with Wilson's disease achieve long-term survival. The postoperative survival time till now is 58 w, 51 w, 48 w, 38 w, 37 w, 36 w, 22 w, 18 w, 10 w and 7 w, respectively. No rejection was detected in the recipients with Wilson's disease, whose liver function and cuprum oxidase level had returned to normal. In this 10 cases, 6 of them has returned or gone to school, 2 of them were discharged from hospital, 2 of them has been recovered smoothly in the hospital due to short time postoperatively. The primary complications after operation including blood vessel, biliary tract, lung and the infection of microbe or virus, were mainly involved in our series.

Conclusion: The process of operation was complicated; the operation technology was exigent and difficult with respect to the safety of the donors and receptors. LRLT has capacious clinical application for there are many unsurpassable advantages, such as plentiful resource and fine quality of graft liver, lower cost of LRLT.
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April 2002

[Application of microsurgical technique to hepatic artery reconstruction in liver transplantation].

Zhonghua Wai Ke Za Zhi 2002 Mar;40(3):205-7

Liver Transplantation Center, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China.

Objective: To explore the value of the microsurgical technique in the reconstruction of hepatic artery.

Methods: From September 2000 to June 2001, we performed liver transplantation for 11 patients including living related liver transplantation (4) and 7 orthotopic liver transplantation (7). Arterial reconstruction was performed under an operating microscope.

Results: No patients developed hepatic arterial thrombosis and serious complication, nor death for multiple organ failure.

Conclusion: Microsurgical technique in reconstruction of the hepatic artery can improve surgical outcome, not only in orthotopic liver transplantation but also in living related liver transplantation.
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March 2002