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Rat orthotopic liver transplantation (ROLT) remains a technically demanding procedure, especially regarding the reconstruction of the suprahepatic vena cava (SHVC). In this study, a new microsuture technique was developed for anastomosis of the SHVC, and a special single-groove cuff and blade-cut stent were introduced. With these modified techniques, we aimed to make a precise anastomosis of the SHVC and to provide optimal cuffs and stents for the reconstruction of the veins and bile ducts. According to different microsuture techniques for the SHVC and different types of cuffs and stents, three ROLT groups were created to compare the operation times and prognoses. Sham operations were performed as controls in the fourth group. The time expenditures with each step were compared among the transplantation groups. Biochemical parameters were tested at the end of a 1-month observation period. The short- and long-term survival rates of the transplantation groups were recorded and compared. Our new microsuture technique was faster than the conventional continuous suture technique for SHVC anastomosis (P < 0.05). The use of a single-groove cuff for reconstruction of the portal vein and the infrahepatic vena cava shortened the anastomotic time (P < 0.05). The use of blade-cut stents resulted in fewer biliary complications and better survival over the short and long terms (P < 0.05). Our new microsuture technique and the single-groove cuffs proved to be a precise method for venous reconstruction which shortened the anhepatic time and the anastomotic time significantly. The blade-cut stents apparently reduced the incidence of biliary complications. In summary, with this precise microsuture technique and delicate cuffs and stents, excellent long-term survival can be achieved easily and stably for ROLT.

Decellularized whole organs represent ideal scaffolds for engineering new organs and/or cell transplantation. Here, we investigate whether decellularized liver scaffolds provide cell-friendly biocompatible three-dimensional (3-D) environment to support the proliferation and differentiation of hepatic progenitor cells. Mouse liver tissues are efficiently decellularized through portal vein perfusion. Using the reversibly immortalized mouse fetal hepatic progenitor cells (iHPCs), we are able to effectively recellularize the decellularized liver scaffolds. The perfused iHPCs survive and proliferate in the 3-D scaffolds in vitro for 2 weeks. When the recellularized scaffolds are implanted into the kidney capsule of athymic nude mice, cell survival and proliferation of the implanted scaffolds are readily detected by whole body imaging for 10 days. Furthermore, epidermal growth factor (EGF) is shown to significantly promote the proliferation and differentiation of the implanted iHPCs. Histologic and immunochemical analyzes indicate that iHPCs are able to proliferate and differentiate to mature hepatocytes upon EGF stimulation in the scaffolds. The recellularization of the biomaterial scaffolds is accompanied with vascularization. Taken together, these results indicate that decullarized liver scaffolds effectively support the proliferation and differentiation of iHPCs, suggesting that decellularized liver matrix may be used as ideal biocompatible scaffolds for hepatocyte transplantation.

// Xiwen Liao 1, * , Chuangye Han 1, * , Wei Qin 1, * , Xiaoguang Liu 2 , Long Yu 3 , Guangzhi Zhu 1 , Tingdong Yu 1 , Sicong Lu 1 , Hao Su 1 , Zhen Liu 1 , Zhiwei Chen 1 , Chengkun Yang 1 , Ketuan Huang 1 , Zhengtao Liu 1 , Yu Liang 1 , Jianlu Huang 4 , Jiahong Dong 5 , Lequn Li 6 , Xue Qin 7 , Xinping Ye 1 , Kaiyin Xiao 1 , Minhao Peng 1 , Tao Peng 1 1 Department of Hepatobiliary Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi Province, China 2 Department of Hepatobiliary Surgery, Affiliated Hospital of Guangdong Medical University, Zhanjiang, 524001, Guangdong Province, China 3 Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450000, Henan Province, China 4 Department of Hepatobiliary Surgery, The Third Affiliated Hospital of Guangxi Medical University, Nanning, 530031, Guangxi Province, China 5 Department of Hepato-Biliary-Pancreatic Surgery, Beijing Tsinghua Changgung Hospital, Beijing, 102218, China 6 Department of Hepatobiliary Surgery, Affiliated Tumor Hospital of Guangxi Medical University, Nanning, 530021, Guangxi Province, China 7 Department of Clinical Laboratory, The First Affiliated Hospital of Guangxi Medical University, Nanning, 530021, Guangxi Province, China * These authors have contributed equally to this work and should be considered co-first authors Correspondence to: Tao Peng, email: pengtaogmu@163.com Keywords: hepatocellular carcinoma, hepatitis B virus, PLCE1, AFP, prognosis Received: October 13, 2016 Accepted: February 22, 2017 Published: March 18, 2017 ABSTRACT Polymorphisms in the phospholipase C epsilon ( PLCE ) 1 gene play a crucial role in the development and progression of several types of cancer. The present study investigated the prognostic significance of PLCE1 gene polymorphisms and expression combined with serum α-fetoprotein (AFP) level in hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC). Single nucleotide polymorphisms were genotyped by sequencing DNA isolated from surgically resected tumor samples of 421 HBV-related HCC patients, and expression profiles were generated based on the GSE14520 dataset. A joint-effects analysis of PLCE1 haplotypes (A rs2274223 C rs3765524 ; G rs2274223 T rs3765524 ) with AFP level stratified at 20 ng/ml showed a significant association with overall survival(OS) of HBV-related HCC patients(log-rank P =0.0003). Patients with AC and GT haplotypes with AFP level ≥ 20 ng/ml had an increased risk of death as compared to those with the AC haplotype and AFP level < 20 ng/ml (adjusted P =0.029 and 0.041, respectively). Patients with the GT haplotype and AFP level < 20 ng/ml also had an increased risk of death, although with a non-significant P value (adjusted P =0.092). Joint-effects analysis of PLCE1 mRNA expression with serum AFP level stratified at 300 ng/ml was significantly associated with HBV-related HCC recurrence and OS. Our results demonstrate that PLCE1 haplotypes (including rs2274223 and rs3765524) and expression combined with serum AFP level may predict postoperative outcome of HBV-related HCC patients.

Gallbladder carcinoma (GBC) is the most common biliary tract malignancy with high mortality. The median survival time is 6 months, and the 5-year survival rate less than 5% for GBC patients. Thus, it is imperative to investigate the molecular mechanisms underlying the pathogenesis of GBC. miR-133a may exert anti-tumor effects on a variety of cancers. However, the role of miR-133a in the pathogenesis of GBC remains unclear. In this study, quantitative real-time polymerase chain reaction (qRT-PCR) showed the miR-133a-3p expression markedly decreased in GBC as compared to adjacent normal tissues. Transient over-expression of miR-133a-3p inhibited the proliferation, migration and invasion abilities of GBC cells. Luciferase activity assay indicated that miR-133a-3p negatively regulated the expression of recombination signal-binding protein Jκ (RBPJ) directly, which is a key downstream transcription factor in the Notch signaling pathway. Moreover, PBPJ expression was up-regulated and negatively related to miR-133a-3p expression in GBC, and silencing of RBPJ achieved the effects as after miR-133a-3p over-expression. RBPJ over-expression could markedly reverse the inhibitory effects of miR-133a-3p on the proliferation, migration and invasion of GBC cells. Our findings indicate that miR-133a-3p acts as a tumor suppressor through directly targeting RBPJ in GBC.

It remains unclear whether hepatectomy for colorectal liver metastasis (CRLM) should be performed as anatomical resection (AR) or nonanatomical resection (NAR). The aim of this study is to compare the short- and long-term outcomes of AR and NAR for CRLM. PubMed, Web of Science, EMBASE and the Cochrane Library were systematically searched to identify eligible studies. Twenty one studies involving 5207 patients were analyzed: 3034 (58.3%) underwent AR procedure and 2173 (41.7%) underwent NAR procedure. The results showed that overall survival (OS, hazard ratio (HR) 1.06, 95% confidence interval (CI) 0.95–1.18) and disease free survival (DFS, HR 1.11, 95% CI 0.99–1.24) did not differ significantly between AR and NAR. Duration of operation, postoperative morbidity and mortality were higher in AR than in NAR. There were no significant differences in blood loss and prevalence rate of postoperative positive margins (OR 0.79, 95% CI 0.37–1.52). Our analysis shows that AR does not seem to bring more prognostic benefits than NAR for the treatment of CRLM, and does seem to be inferior to NAR in terms of duration of operation, incidence of postoperative morbidity and mortality.

Our data indicate that in the context of HCC, miR-214 acts as a putative tumour suppressor by targeting UCP2 and defines a novel mechanism of regulation of UCP2., Gemcitabine (GEM), a commonly used chemotherapeutic agent in hepatocellular carcinoma (HCC) patients, uses oxidative stress induction as a common effector pathway. However, GEM alone or in combination with oxaliplatin hardly renders any survival benefits to HCC patients. We have recently shown that this is part due to the overexpression of the mitochondrial uncoupling protein 2 (UCP2) that in turn mediates resistance to GEM in HCC patients. However, not much is known about regulatory mechanisms underlying UCP2 overexpression in HCC. Differential protein expression in HCC cell lines did not show a concomitant change in UCP2 transcript level, indicating post-transcriptional or post-translational regulatory mechanism. In situ analysis revealed that UCP2 is a putative target of miR-214. miR-214 expression is significantly down-regulated in HCC patient samples as compared with normal adjacent tissues and in cell line, human hepatoblastoma cells (HuH6), with high UCP2 protein expression. We demonstrated using miR-214 mimic and antagomir that the miRNA targeted UCP2 expression by directly targeting the wild-type, but not a miR-214 seed mutant, 3’ UTR of UCP2. Overexpression of miR-214 significantly attenuated cell proliferation. Finally, analysis in 20 HCC patients revealed an inverse correlation in expression of UCP2 and miR-214 (Pearson's correlation coefficient, r=−0.9792). Cumulatively, our data indicate that in the context of HCC, miR-214 acts as a putative tumour suppressor by targeting UCP2 and defines a novel mechanism of regulation of UCP2.

Acute liver failure (ALF) is a reversible disorder that is associated with an abrupt loss of hepatic mass, rapidly progressive encephalopathy and devastating complications. Despite its high mortality, an emergency liver transplantation nowadays forms an integral part in ALF management and has substantially improved the outcomes of ALF. Here, we report the case of a 32-year-old female patient who was admitted with grade IV hepatic encephalopathy (coma) following drug-induced ALF. We performed an emergency auxiliary partial orthotopic liver transplantation with a "high risk" graft (liver macrovesicular steatosis approximately 40%) from a living donor. The patient was discharged on postoperative day 57 with normal liver function. Weaning from immunosuppression was achieved 9 mo after transplantation. A follow-up using CT scan showed a remarkable increase in native liver volume and gradual loss of the graft. More than 6 years after the transplantation, the female now has a 4-year-old child and has returned to work full-time without any neurological sequelae.

To describe the clinicopathological characteristics and prognosis of intrahepatic biliary cystic tumor (IBCT) and compare the difference betwwen male and female patients.11 male and 45 female patients with histological confirmed IBCT from January 2003 to April 2014 were envolved. clinical characteristics, laboratory examination, radiological features, treatment method and prognosis were analyzed.The mean age of male patients with IBCT was significantly older than that of female patients (54.9±10.1 years vs 46.2±12.9 years, P=0.041). There were no significant differences in radiological features between male and female patients except for mural nodules (10/11 vs 15/45, P=0.001). Intrahepatic biliary cystadenocarcinoma (IBC) was more common in male patients than female patients (8/11 vs 16/45, P=0.041). Male patients had worse prognosis than female patients (P=0.014). 1-year, 3-year, and 5-year survival of male patients was 90.9%,45.5%,30.3%, respectively. However, 1-year, 3-year, and 5-year survival of female patients was 94.8%,82.7%,78.6%.Male patients had low incidence of IBCT, and were more common for IBC. Even with complete resection, the long-term survival was still poor, close follow-up were recommended especially for male patients.

Serological platform using three monoclonal antibodies for identification of Calla lily chlorotic spot virus (CCSV), Tomato zonate spot virus (TZSV) and Tomato necrotic spot associated virus (TNSaV). (DOCX 12 kb)

Primers used for amplification of complete or truncated open reading frames of nucleocapsid (N) gene of Tomato zonate spot virus (TZSV) 13YV639 isolate. (DOCX 14 kb)

Summary of recombination events in different full-length TSWV L isolates using RDP4. NS: not significant. (DOCX 33Â kb)

SPIRIT 2013 Checklist: recommended items to address in a clinical trial protocol and related documents. (DOC 123 kb)

Primers used in this study. Primers were designed based on previously reported TSWV sequences. W: A/T, M: A/C, R: G/A, Y: C/T. (DOCX 15Â kb)

Summary of recombination events in different full-length TSWV L isolates using RDP4. NS: not significant. (DOCX 31Â kb)

TSWV isolates used in this study. TSWV isolates from different countries use different abbreviations as followed, China (CL, CM and CS), South Korea (KL, KM and KS), Japan (JL, JM, JS), USA (UL, UM, US), Brazil (BL, BM, BS), Italy (IL, IM IS), Australia (AM), Spain (SM, SS), Germany (GS), Bulgaria (BuS). CS: Complementary sequence; N/A: Not analyzed. (DOCX 35Â kb)

Comparison of the serological relationship of Calla lily chlorotic spot virus (CCSV), Tomato zonate spot virus (TZSV) and Tomato necrotic spot associated virus (TNSaV). The previously reported monoclonal antibodies MAb-CCSV-NP [24] (A and B) and MAb-WNSs [25] (C and D) were used to react with the crude leaf extracts of Nicotiana benthamiana plants separately infected with CCSV, TZSV, TNSaV or Watermelon silver mottle virus (WSMoV) in indirect enzyme-linked immunosorbent assay (A and C) and immunoblotting (B and D). The crude extract of a healthy N. benthamiana leaf (H) was used as the negative control. The plant ribulose bisphosphate carboxylase/oxygenase (rubisco) of Ponceau S staining is shown under immunoblotting to indicate the loading quantity. MAb-CCSV-NP was used at a 10â 4 dilution. MAb-WNSs was used at a 10â 3 dilution. (DOCX 124 kb)

Background Tomato spotted wilt virus (TSWV) and Tomato zonate spot virus (TZSV) are the two dominant species of thrip-transmitted tospoviruses, cause significant losses in crop yield in Yunnan and its neighboring provinces in China. TSWV and TZSV belong to different serogroup of tospoviruses but induce similar symptoms in the same host plant species, which makes diagnostic difficult. We used different electron microscopy preparing methods to investigate clustering and cellular distribution of TSWV and TZSV in the host plant species. Results Negative staining of samples infected with TSWV and TZSV revealed that particles usually clustered in the vesicles, including single particle (SP), double particles clustering (DPC), triple particles clustering (TPC). In the immunogold labeling negative staining against proteins of TZSV, the antibodies against Gn protein were stained more strongly than the N protein. Ultrathin section and high pressure freeze (HPF)-electron microscopy preparations revealed that TSWV particles were distributed in the cisternae of endoplasmic reticulum (ER), filamentous inclusions (FI) and Golgi bodies in the mesophyll cells. The TSWV particles clustered as multiple particles clustering (MPC) and distributed in globular viroplasm or cisternae of ER in the top leaf cell. TZSV particles were distributed more abundantly in the swollen membrane of ER in the mesophyll cell than those in the phloem parenchyma cells and were not observed in the top leaf cell. However, TZSV virions were mainly present as single particle in the cytoplasm, with few clustering as MPC. Conclusion In this study, we identified TSWV and TZSV particles had the distinct cellular distribution patterns in the cytoplasm from different tissues and host plants. This is the first report of specific clustering characteristics of tospoviruses particles as well as the cellular distribution of TSWV particles in the FI and globular viroplasm where as TZSV particles inside the membrane of ER. These results indicated that tospoviruses particles possessed specific and similar clustering in the saps of diseased plants. Furthermore, the results of this study will also provide a basis for further study on the tospoviruses assembling, maturation and movement.

Summary of recombination events in different full-length TSWV L isolates using RDP4. NS: not significant. (DOCX 24Â kb)

The aim of this study was to evaluate the association between activating enhancer binding protein 4 (AP-4) tissue expression and patient prognosis in hepatocellular carcinoma (HCC). The levels of AP-4 mRNA and protein in tumor and para-tumor tissue were evaluated in 30 HCC cases by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. Additionally, AP-4 protein expression in 112 HCC was analyzed by immunohistochemistry. The correlation of AP-4 expression and patients’ clinicopathological parameters was evaluated. Survival analysis was performed using the Kaplan–Meier method and Cox’s proportional hazards model. By RT-PCR and Western blot, the levels of AP-4 mRNA and protein were significantly higher in HCC, compared to that in para-tumor tissue (p

The cell growth, development, and regeneration of tissue and organ are associated with a large number of gene regulation events, which are mediated in part by transcription factors (TFs) binding to cis-regulatory elements involved in the genome. Predicting the binding affinity and inferring the binding specificity of TF–DNA interactions at the genomic level would be fundamentally helpful for our understanding of the molecular mechanism and biological implication underlying sequence-specific TF–DNA recognition. In this study, we report the development of a combination method to characterize the interaction behavior of a 11-mer oligonucleotide segment and its mutations with the Gcn4p protein, a homodimeric, basic leucine zipper TF, and to predict the binding affinity and specificity of potential Gcn4p binders in the genome-wide scale. In this procedure, a position-mutated energy matrix is created based on molecular modeling analysis of native and mutated Gcn4p–DNA complex structures to describe the position-independent interaction energy profile of Gcn4p with different nucleotide types at each position of the oligonucleotide, and the energy terms extracted from the matrix and their interactives are then correlated with experimentally measured affinities of 19 268 distinct oligonucleotides using statistical modeling methodology. Subsequently, the best one of built regression models is successfully applied to screen those of potential high-affinity Gcn4p binders from the complete genome. The findings arising from this study are briefly listed below: (i) The 11 positions of oligonucleotides are highly interactive and non-additive in contribution to Gcn4p–DNA binding affinity; (ii) Indirect conformational effects upon nucleotide mutations as well as associated subtle changes in interfacial atomic contacts, but not the direct nonbonded interactions, are primarily responsible for the sequence-specific recognition; (iii) The intrinsic synergistic effects among the sequence positions of oligonucleotides determine Gcn4p–DNA binding affinity and specificity; (iv) Linear regression models in conjunction with variable selection seem to perform fairly well in capturing the internal dependences hidden in the Gcn4p–DNA system, albeit ignoring nonlinear factors may lead the models to systematically underestimate and overestimate high- and low-affinity samples, respectively.

Laparoscopic surgery for confirmed infected pancreatic necrosis (IPN) represents a relatively new solution. There are no studies comparing the outcomes of laparoscopic and open surgery for patients with IPN. The aims of this study were to investigate the feasibility of laparoscopic management for patients with IPN and to compare the outcomes of laparoscopic and open surgery.Seventy-six patients with IPN who underwent open surgery (Open-group) or laparoscopic surgery (Lap-group) were retrospectively reviewed. Demographic data, white blood cell count, and APACHE II score upon admission, operative findings, major complications, and mortality were compared between the Open-group and the Lap-group. The Lap-group was further divided into two subgroups (early and late), and the operative difficulty was compared between the two subgroups.There were no significant differences between the Open-group and the Lap-group with respect to demographic data, white blood cell count, and APACHE II score. Although the mean operative time was significantly shorter in the Open-group than in the Lap-group, the estimated blood loss was significantly greater in the Open-group than in the Lap-group, as was the rate of complications. The mean postoperative hospital stay in the Open-group was significant longer than in the Lap-group, too. In the Lap-group, the mean operating time, estimated blood loss, and conversion rate in the early subgroup were significantly lower than in the late subgroup.Laparoscopic necrosectomy and the placement of an intermittent irrigation and continuous suction drainage system for IPN is feasible, effective, and of minimal invasiveness. The late laparoscopic necrosectomy is relatively difficult.

To investigate the method and effect of percutaneous nephroscopic necrosectomy (PNN) for post-operatively resident infection of severe acute pancreatitis (SAP).Data of the 15 SAP patients with post-operatively resident infection treated by PNN from June 2008 to December 2014 in Chinese People's Liberation Army General Hospital were reviewed. Twelve of the patients underwent the laparotomy within 1 week, 1 in 3(rd) week, 1 in 4(th) week and the other one on the 127(th) day. All of the referrals firstly received (multi-)percutaneous catheter drainage (PCD), and then PNN operation according to the disease, followed by continuous irrigation-drainage.Eleven patients were healed after received only one PNN operation, 2 patients for twice, 1 for three times and 1 for four times. The average post-operative time of hospital stay was 66.2 days (10-223 days). The complications after operation contained colon fistula (n = 1), abdominal hemorrhage (n = 1), pancreatic pseudocyst (n = 1), severe pulmonary infection (n = 1). Three patients eventually died.Percutaneous nephroscopic necrosectomy is a minimally invasive approach which could prevent the complicated re-laparotomy operation, result in less complication. It is an ideal method for treating SAP patients with post-operatively resident infection.

To describe the treatment and prognosis of solid pseudopapillary neoplasms (SPN) with metastases or recurrence.The clinical date of 24 patients with histological confirmed SPN with metastases or recurrence from January 2000 to April 2014 were retrospectively analyzed. There were 22 females and 2 males, with mean age of (36 ± 16) years. Fourteen patients had local recurrence or metastasis after surgery, with a mean time of recurrence (44 ± 29) months. Ten patients were defined SPN with distant metastasis at first admission. Nineteen patients underwent surgical resection, among them, 11 patients received complete resection. Nine cases underwent chemotherapy. Kaplan-Meier method was used to identify prognostic factors.Twenty-four patients were followed-up, 9 patients died. Median survival time was 47 months, and 1-year, 3-year, and 5-year survival was 91.7%, 65.1%, 49.6%, respectively. Age (χ(2) = 6.858, P = 0.009), primary tumor diameter (χ(2) = 4.322, P = 0.038), extrahepatic metastasis (χ(2) = 5.279, P = 0.022) and complete resection of metastases and recurrence (χ(2) = 4.666, P = 0.031) were important prognostic factors for survival (P0.05).For SPN with metastases or recurrence, good prognosis can also obtain after complete resection. Age, primary tumor diameter, extrahepatic metastasis and complete resection of metastases and recurrence are influence factors on prognosis of patients.

Viral cirrhotic primary liver cancer (VCPLC) is characterized by intra-hepatic metastasis (IHM) at early stage. By contrast, simple primary liver cancer without cirrhosis (SPLC) has high extra-hepatic metastasis (EHM) rate. Historically, studies on mechanism of metastasis of PLC mainly focused on biological behaviors of cancer cells, however tumor microenvironment is always neglected, especially blood flow dynamics and Kupffer cells. In SPLC, few damages of hepatic lobules occur, to the contrary, VCPLC is characterized by obvious hepatic fibrosis which results in varieties of blood flow disturbances, such as central lobular vein obstruction, hepatic artery-portal vein fistula (HAPVF), and portal vein countercurrent (PVC). Traditionally, Kupffer cells in liver are regarded as powerful defenses against PLC and metastasis lesions in liver, however, recent studies show under long term ischemic-hypoxic stress, Kupffer cells can secrete varieties of pro-inflammatory mediators to promote fibrosis or even cancerogenesis. Tumor associated macrophages (TAMs) are a group of macrophages living in ischemic tumor lesions which can promote tumor progression and metastasis. Based on the totally different metastasis routes and significantly different tumor microenvironment of VCPLC and SPLC, we hypothesize: (1) unlike the normal Kupffer cells in non-cirrhosis liver, Kupffer cells of VCPLC, a group of dominant hepatic resident macrophages living in extremely cirrhotic ischemic-hypoxic sinusoids, might not act as fighters against cancer cells, what is worse, ischemic-hypoxic situation might incite Kupffer cells to secrete cytokines and chemokines, just like TAMs, to promote tumor growth and metastasis; (2) the blood flow disturbances, not the behaviors of cancer cells, are the main force to determine the different metastasis preferences. In VCPLC, central lobular vein is mechanically compressed by cirrhosis nodes, so cancer cells cannot go extra-hepatically, instead, the hepatic artery-portal vein fistula (HAPVF) and portal vein countercurrent (PVC) load cancer cells intra-hepatically. In SPLC, cancer cells can run directly extra-hepatically through central lobular vein fluently. In short, the different blood flow dynamics between these two PLC is the predominant factor to determine the preferences of metastasis route. In VCPLC, the Kupffer cells should not be the powerful defenses against tumor and metastasis lesions, instead, they should be tamed to be some TAMs by long term ischemic-hypoxic stress and tumor derived factors to play important roles in tumorigenesis or even metastasis, so Kupffer cells can be a new target for preventing hepatocancerogenesis and metastasis.

Ethanol preconditioning (EtOH-PC) refers to a phenomenon in which cerebral, intestinal, and myocardial tissues are protected from the deleterious effects of ischemia/reperfusion (I/R) by prior ingestion of ethanol at low to moderate levels. Whether EtOH-PC can offer protective effects against hepatic I/R injury and whether these effects are associated with inhibition of complement activation were investigated.Male SD rats were divided into four groups, i.e., sham operation, ethanol control, IR, and ethanol-pretreatment I/R (EIR) groups. EtOH-PC was induced by gavaging rats with 40% ethanol at a dose of 5 g/kg body weight 24 h prior to experiment. Animal survival rate was compared. Liver function, hepatic MDA level, plasma complement C3 level, and serum hemolytic activity were determined. Histologic changes and complement C3 deposition in liver section were examined. Expression of liver complement 3 mRNA was analyzed by quantitative real-time -PCR.The 14-d survival rates were remarkably higher in the EIR groups than in the corresponding IR groups when hepatic ischemia time was 110, 120, and 130 min. Serum ALT, AST, IL-1β, and liver tissue MDA were significantly lower, and histopathologic changes significantly milder in the EIR group than in the IR group (P0.05). Compared with the IR group, both the reduction in CH50 and plasma C3 were significantly suppressed, and the staining of C3 in liver tissue significantly reduced in the EIR group. There were no significant differences of hepatic C3 mRNA among four groups.Ethanol preconditioning reduces hepatic I/R injury, and the effect is associated with inhibition of complement activation.

Highlights • Anatomical resection of S8vent is generally considered a demanding procedure. • There are no accurate methods to identify the demarcation line inside the liver by far. • We proposed a new method to divide the intersegmental plane by exposing and following the small tributaries of hepatic vein near the liver surface. • This method is time-consuming, but it is an effective way to divide the intersegmental plane., INTRODUCTION Anatomical resection of the ventral part of Segment VIII (S8vent) is demanding and there are no accurate methods to identify the demarcation line inside the liver. The current authors have proposed a method to solve the problem. PRESENTATION OF CASE The tumor was located in the S8vent and was 4 cm in size. One tributary of the middle hepatic vein (MHV), designated V8i, was running between S8vent and the dorsal part of Segment VIII (S8dor). Another tributary of the MHV, designated V8-5i, was running between S8vent and the ventral part of S5 (S5vent). About 5 ml of indigo carmine dye was injected into the proximal part of P8vent. After the small tributary of V8-5i was exposed, it was followed all the way to the main trunk of the MHV. The portal pedicle of S8vent was then ligated and divided. Next, the V8i was gradually exposed from the distal MHV to its trunk. DISCUSSION A recent study showed that the subsegmental border visualized between the ventral and dorsal region always coincided with the plane of V8i, so the subsegmental plane can be divided along with V8i by preserving the very small tributaries near the liver surface and following them to determine where they meet as they run into V8i. Also, the landmark vein of V8-5i in the transverse S8–S5 intersegmental plane was determined for the first time. CONCLUSION Proposed here is a more accurate method of dividing the liver parenchyma along the intersegmental and intersubsegmental demarcation.

Background Radiofrequency ablation (RFA) is a relatively new treatment for hypersplenism. The results of a randomized clinical trial comparing RFA and splenectomy with 5 years of follow-up are reported. Methods Fifty-seven patients with hypersplenism due to liver cirrhosis were assigned randomly (in a 1 : 2 ratio) to splenectomy (19 patients) or RFA (38). The RFA group was subdivided according to the percentage of the spleen ablated: less than 50 per cent (9 patients), 50–70 per cent (18) or over 70 per cent (11). Routine blood tests were performed before and after operation, and total spleen volume and ablated volume were measured by contrast-enhanced computed tomography. The primary endpoint of the trial was recurrence of hypersplenism, assessed as platelet and white cell counts, at 5 years after surgery. Results White cell and platelet counts increased rapidly after intervention in both groups. By 36 months after operation peripheral platelet and white cell counts had decreased significantly in the RFA group compared with the splenectomy group, and declined to baseline levels by 48 months. Hypersplenism recurred after 6 months in patients with less than 50 per cent of the spleen ablated. Blood cell count in the splenectomy group and in patients with more than 50 per cent of the spleen ablated decreased with time after operation, but to levels that remained significantly higher than those before operation (P < 0·050). Splenic volume reached its nadir 12 months after RFA and then increased with time. Conclusion Splenic RFA represents an attractive alternative treatment for hypersplenism induced by liver cirrhosis, particularly when more than 50 per cent of the spleen is ablated.

In this study, we examined the effects of LY52, a caffeoyl pyrrolidine derivative designed to fit the S'1 active pocket of gelatinases, on the expressions of matrix metalloproteinases and invasion abilities of hepatocellular carcinoma cells.The effects of LY52 on the proliferations of HepG2 (hepatitis B virus (HBV) negative) and HepG2.2.15 (HBV-producing) cells were detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Gelatin zymography was used to detect the effects of LY52 on matrix metalloproteinases expressions and Western blot was used to detect matrix metalloproteinase-2 expressions. Transwell chamber assay was used to detect the effects of LY52 on invasion of the cells.Gelatin zymography and Western blot showed that matrix metalloproteinase-2 expressions were inhibited by LY52 in a dose-dependent manner, and inhibitory rates of LY52 on HepG2 cells were higher than on HepG2.2.15 cells. Transwell chamber showed that LY52 could significantly inhibit the invasion of both cells, although the inhibitory effects of LY52 on HepG2.2.15 cells were was not as obvious as on HepG2 cells.These results suggested that LY52 might inhibit the invasion of hepatocellular carcinoma cells by suppressing matrix metalloproteinase-2, although the inhibitory effects of LY52 on HBV-negative cells were more obvious than that of HBV-infected cells.

Background and Aims: Src family kinases have been suggested to be associated with the metastasis of tumors, but their related mechanisms remain unclear. The aims of the present study were to assess the possible mechanisms by which the inhibition of Fyn activation regulates pancreatic cancer metastasis. Methods: We examined the expressions of Fyn in human pancreatic cancer tissues by immunohistochemistry and systematically investigated the relationship between Fyn expression and pancreatic cancer metastasis. A nude mouse xenograft model induced by BxPC3 cells with or without the inhibition of Fyn activation was used to explore the effect of the inhibition of Fyn on metastasis in vivo. Methyl thiazolyl tetrazolium and terminal deoxynucleotidyl transferase-labeling assays were used to examine the effect of the inhibition of Fyn on the cell proliferation of BxPC3 pancreatic cancer cells in vitro. Reverse transcription polymerase chain reaction and Western blot analysis were performed to explore the possible mechanism of Fyn-induced metastasis. Results: We found that the upregulation of Fyn expression was correlated with human pancreatic cancer metastasis. In BxPC3 pancreatic cancer cells, the inhibition of Fyn activation by kinase-dead Fyn transfection decreased liver metastasis in nude mice. Further analyses showed that Fyn activity modulated pancreatic cell metastasis through the regulation of proliferation and apoptosis. Conclusions: Our results suggest a possible mechanism by which Fyn activity regulates cell proliferation and apoptosis that exerts an effect on pancreatic cancer metastasis.

Two new quassinoids, javanicolide E (1) and javanicolide F (2), along with fifteen known C-20 quassinoids were isolated from the seeds of Brucea javanica (L.) Merr. The antitobacco mosaic virus (TMV) activity of these quassinoids was screened by the conventional half-leaf and leaf-disk method along with Western blot analysis. All of the seventeen quassinoids showed potent anti-TMV activity. Among them, eight compounds, brusatol (3), bruceine B (4), bruceoside B (5), yadanzioside I (6), yadanzioside L (7), bruceine D (8), yadanziolide A (9), and aglycone of yadanziolide D (17), showed strong antiviral activities, with IC(50) values in the range of 3.42-5.66 microM, and were much more effective than the positive control, ningnanmycin (IC(50) = 117.3 microM). The antiviral structure-activity relationships of quassinoids against TMV were also discussed.

Caveolin-1 is the major structural protein in caveolae, implicated in oncogenesis and angiogenesis. The connections between caveolin-1 and progression and angiogenesis of hepatocellular carcinoma (HCC) is still not clear. Thus we investigated the relationship of caveolin-1 expression, vascular endothelial growth factor (VEGF) expression, microvessel density (MVD), and unpaired artery (UA) with the clinicopathologic features of patients with HCC. Formalin-fixed, paraffin-embedded tissue sections of HCC from 75 patients who had undergone an initial hepatectomy were stained immunohistochemically with specific antibodies against caveolin-1, VEGF, CD34 and alpha-SMA. The levels of caveolin-1, VEGF, MVD and UA were correlated with the clinicopathologic variables, and tissue sections were also analyzed by dual-label immunofluorescence. We found that increased expression of caveolin-1 was associated with metastasis and with a worse prognosis of HCC. Caveolin-1 expression correlates positively with VEGF, MVD and UA. These results suggest that caveolin-1 may play an important role in the progression of HCC and angiogenesis.

Purpose: Interaction between tumor cells and surrounding stromal fibroblast (SF) plays a critical role in tumor growth and invasion. The aim of the study is to determine the role of SF in regulating the invasive behaviors of pancreatic cancer by evaluating the mode of SF activating the urokinase plasminogen activator (uPA)-plasmin-matrix metalloproteinase (MMP)-2 cascade. Experimental Design: The expression patterns of uPA, MMP-2, and uPA receptor (uPAR) in human metastatic pancreatic cancer were analyzed by immunohistochemistry and the roles of SF in activation of the uPA-plasmin-MMP-2 cascade were evaluated by coculturing pancreatic cancer cell lines with SF. Results: uPA expression and fibroblastic uPAR expression were correlated with liver metastasis of human pancreatic cancer. MMP-2 rather than MMP-9 was activated in the metastatic pancreatic cancer. In the in vitro culture system, the coculture of peritumor fibroblasts with metastatic pancreatic cancer BxPc3 cells resulted in activation of MMP-2 and up-regulation of uPAR expression. In this coculture system, the uPA-plasminogen cascade was involved in MMP-2 activation. This activation required a direct interaction between SF and cancer cells. In the coculture system, intergrin α6β1 expression was increased in BxPc3 cells, and blocking the function of integrin α6β1 decreased the activation of uPA and MMP-2. This suggests that interaction between integrins of cancer cells and the uPARs of the SF might be involved in the activation of the uPAR-uPA-MMP-2 cascade. Conclusion: Our results suggest that SF plays a role in promoting pancreatic cancer metastasis via activation of the uPA-plasminogen-MMP-2 cascade.

The virus isolate 2009-GZT, collected from tomato in Guizhou province of China, was identified as a new member of the genus Tospovirus based on its S RNA sequence. Because its provisional name, "tomato necrotic spot virus" (TNSV), was identical to an already existing member of the genus Ilarvirus, 2009-GZT was renamed "tomato necrotic spot-associated virus" (TNSaV). In this study, the full-length sequences of the genomic M and L RNAs of TNSaV were determined and analyzed. The M RNA has 4,773 nucleotides (nt), encoding the NSm protein of 309 aa (34.4 kDa) in the viral (v) strand and the glycoprotein precursor (Gn/Gc) of 1123 aa (128 kDa) in the viral complementary (vc) strand. The NSm and Gn/Gc of TNSaV share the highest aa sequence identity (86.2 % and 86.9 %, respectively) with those of tomato zonate spot virus. The L RNA contains 8,908 nt and codes for the putative RNA-dependent RNA polymerase (RdRp) of 2885 aa (332 kDa) in the vc strand. The RdRp of TNSaV shares the highest aa sequence identity (85.2 %) with that of calla lily chlorotic spot virus (CCSV). Serological assays showed that TNSaV cross-reacts with rabbit antisera against watermelon silver mottle virus (WSMoV) NP and CCSV NP, indicating that TNSaV is a member of the WSMoV serogroup.

To analyze the clinicopathological characteristics and the factors affecting the prognosis of intrahepatic biliary cystadenocarcinoma (IBC).Forty-six patients with histologically confirmed IBC from January, 2000 to April, 2014 were included. The clinical characteristics of the patients with IBC were compared with those of 58 patients with intrahepatic biliary cystadenoma (IBCA). Kaplan-Meier analysis was used to identify the factors affecting the prognosis of IBC.The 46 patients with IBC included 15 men and 31 women with a mean age of 57.0 ± 10.5 years. Compared with the patients with IBCA, IBC patients had an older mean age (57.0 ± 10.7 vs 44.3 ± 15.3 years, P=0.03) and a greater proportion of male patients (15/46 vs 8/58, P=0.02). The differential diagnosis between IBC and IBCA was difficult on the basis of preoperative laboratory and imaging findings. The median overall survival of IBC patients was 56 months with 1-, 3-, and 5-year survival rates of 85.9%, 65.2%, and 47.7%, respectively. Gender, surgical approach, tumor growth pattern and distant metastasis were all significant prognostic factors for the overall survival of the patients.IBC is a rare cystic lesion occurring primarily in middle-aged men. Complete resection is recommended for curative treatment and close follow-up is essential especially for male patients and patients with tumors exhibiting an invasive growth.

Background Destructive diseases caused by Tomato spotted wilt virus (TSWV) have been reported associated with many important plants worldwide. Recently, TSWV was reported to infect different hosts in China. It is of value to clone TSWV isolates from different hosts and examine diversity and evolution among different TSWV isolates in China as well as worldwide. Methods RT-PCR was used to clone the full-length genome (L, M and S segments) of three new isolates of TSWV that infected different hosts (tobacco, red pepper and green pepper) in China. Identity of nucleotide and amino acid sequences among TSWV isolates were analyzed by DNAMAN. MEGA 5.0 was used to construct phylogenetic trees. RDP4 was used to detect recombination events during evolution of these isolates. Results Whole-genome sequences of three new TSWV isolates in China were determined. Together with other available isolates, 29 RNA L, 62 RNA M and 66 RNA S of TSWV isolates were analyzed for molecular diversity, phylogenetic and recombination events. This analysis revealed that the entire TSWV genome, especially the M and S RNAs, had major variations in genomic size that mainly involve the A-U rich intergenic region (IGR). Phylogenetic analyses on TSWV isolates worldwide revealed evidence for frequent reassortments in the evolution of tripartite negative-sense RNA genome. Significant numbers of recombination events with apparent 5′ regional preference were detected among TSWV isolates worldwide. Moreover, TSWV isolates with similar recombination events usually had closer relationships in phylogenetic trees. Conclusions All five Chinese TSWV isolates including three TSWV isolates of this study and previously reported two isolates can be divided into two groups with different origins based on molecular diversity and phylogenetic analysis. During their evolution, both reassortment and recombination played roles. These results suggest that recombination could be an important mechanism in the evolution of multipartite RNA viruses, even negative-sense RNA viruses. Electronic supplementary material The online version of this article (doi:10.1186/s12985-015-0457-3) contains supplementary material, which is available to authorized users.

Hepatic hemangioma, a common benign tumor of the liver, has caused great threats to the public health. Most patients have an excellent prognosis because of the benign nature of hemangioma. On some occasions, spontaneous rupture of hepatic hemangioma is rarely observed in patients. Therefore, a majority of scholars propose that surgery should be restricted to specific situations. In this case, we presented a patient with spontaneous rupture of hepatic hemangioma in our hospital. After a literature review, we summarized the management and outcome of the patient, which revealed the survival rates of the patients are satisfactory.

Our results suggest a critical role for mitochondrial uncoupling in gemcitabine (GEM) resistance in hepatocellular carcinoma (HCC) cell lines. Hence, synergistic targeting of uncoupling protein 2 (UCP2) in combination with other chemotherapeutic agents might be more potent in HCC patients., Oxidative stress induction is a common effector pathway for commonly used chemotherapeutic agents like gemcitabine (GEM) in hepatocellular carcinoma (HCC) patients. However, GEM alone or in combination with oxiplatin hardly renders any survival benefits to HCC patients. Mitochondrial uncoupling protein 2 (UCP2) is known to suppress mitochondrial reactive oxygen species (ROS) generation, thus mitigating oxidative stress-induced apoptosis. We demonstrate in the present study, using a panel of HCC cell lines that sensitivity to GEM in HCC well correlate with the endogenous level of UCP2 protein expression. Moreover, ectopic overexpression of UCP2 in a HCC cell line with low endogenous UCP2 expression, HLE, significantly decreased mitochondrial superoxide induction by the anti-cancer drug GEM. Conversely, UCP2 mRNA silencing by RNA interference in HCC cell lines with high endogenous UCP2 expression significantly enhanced GEM-induced mitochondrial superoxide generation and apoptosis. Cumulatively, our results suggest a critical role for mitochondrial uncoupling in GEM resistance in HCC cell lines. Hence, synergistic targeting of UCP2 in combination with other chemotherapeutic agents might be more potent in HCC patients.

Surgery has passed through an intuitive and an empirical era and has now entered its modern phase. The enormous progress in biomedicine, the rise of evidence-based medicine and the consensus on the need for humanistic patient care, have laid a foundation for a new surgical paradigm. Based on a series of studies and practices, we advocated the concept of 'Precision Surgery'. It covers the entire operation-centered surgical practice. The strategy of precision surgery is to seek a balance of maximizing the removal of the target lesion, while maximizing the functional liver remnant and minimizing surgical invasiveness. We propose that the concept of precision surgery should be considered for wider application within liver surgery and various surgical fields, and finally realize the multi-objective optimization with certainty-based practice to ensure maximized recovery for each patient.

To analyze the surgical management of incidental gallbladder cancer (IGBC) discovered during or after laparoscopic cholecystectomy (LC) and to evaluate the associated factors of survival.A retrospective analysis of patients with IGBC between January 2002 and December 2013 was performed. A total of 10 080 consecutive patients underwent LC operation for presumed gallbladder benign disease in Chinese People's Liberation Army General hospital. And among them, 83 patients were histologically diagnosed as IGBC. Data covering clinical characteristics, surgery records, local pathological stage, histological features and factors for long term survival were reviewed. The survival analysis was performed using Kaplan-Meier method, and the results were examined using the log-rank test.For multivariate statistical analyses of prognostic factors, a Cox proportional hazards model was carried out.A total of 83 patients with IGBC:68.7% females (57/83), median age of 61 years (range 34-83 years). There were 47 cases accepted the initial simple LC, 18 cases converted to open extended radical cholecystectomy, 16 cases with radical second resection, and 2 cases with re-laparotomy; the 5-year survival rates for each group were 89.4%, 38.9%, 87.5%, and 0, respectively. The 5-year survival rates in T1a, T1b, T2, and T3 stage patients were 95.7% (22/23), 90.0% (18/20), 75.0% (15/20), and 40.0% (8/20), respectively. Univariate analysis for prognostic factors associated with cancer-specific death showed that depth of invasion, lymph-node status, vascular or neural invasion, tumor differentiation, extent of resection, bile spillage during prior LC and type of surgery were statistically significant.In multivariate analysis, depth of invasion, extent of resection and bile spillage were the most important prognostic factors related to both cancer-specific mortality and disease relapse (P0.05).Simple LC is appropriate for T1a patients with clear margin and unbroken gallbladder. An extended radical resection in patients with T1b or more is highly recommended, and provided as a potentially curative R0 resection only if it is necessary.

To evaluate the outcome of patients with irresectable hilar cholangiocarcinoma undergoing orthotopic liver transplantation (OLT) and to identify the prognostic factors that could influence survival.The data of 18 patients who underwent OLT for irresectable hilar cholangiocarcinoma between June 2003 and October 2010 were analyzed retrospectively. There were 12 male and 6 female cases with median of 52 years(range from 34 to 65 years).Fifteen patients underwent modified piggyback liver transplantation, 2 patients underwent classical orthotopic liver transplantation and 1 patient underwent living donor liver transplantation. Data were evaluated regarding tumor size, pathologic stage, overall survival, recurrence rates and prognostic factors.OLT with lymphadenectomy was received by 18 patients with hilar cholangiocarcinoma. Median time until tumor recurrence was 20.5 months(range from 6.0 to 33.0 months). Seventeen patients died during follow-up.Of these, 14 patients died from recurrent or metastatic diseases, 2 patients died from multiple organ dysfunction syndrome during peri-operative period, and one patient died from other cause. The median survival time was 29.5 months(range from 3.0 to 84.0 months). The overall survival rate and recurrence-free survival rate at 1, 3, and 5 year were 16/18, 8/18, 1/18 and 13/18, 2/18, 1/18, respectively.Lymph node metastases had a statistically significant negative impact on overall survival. The 1, 3, and 5 year survival rates were 6/7, 1/7,0 and 10/11, 7/11, 1/11 (P0.05) in lymph node-positive and lymph node-negative patients.Acceptable survival rates can be achieved by OLT for irresectable hilar cholangiocarcinoma without lymph node metastases.Strict patient selection plus multimodal chemoradiation therapy prior to OLT are recommend for patients with lymph node metastases.

Background: Emerging tospoviruses cause significant yield losses and quality reduction in vegetables, ornamentals, and legumes throughout the world. So far, eight tospoviruses were reported in China. Tomato fruits displaying necrotic and concentric ringspot symptoms were found in Guizhou province of southwest China. Finding: ELISA experiments showed that crude saps of the diseased tomato fruit samples reacted with antiserum against Tomato zonate spot virus (TZSV). Electron microscopy detected presence of quasi-spherical, enveloped particles of 80–100 nm in such saps. The putative virus isolate was designated 2009-GZT. Mechanical back-inoculation showed that 2009-GZT could infect systemically some solanaceous crop and non-crop plants including Capiscum annuum, Datura stramonium, Nicotiana benthamiana, N. rustica, N. tabacum and Solanum lycopersicum. The 3012 nt full-length sequence of 2009-GZT S RNA shared 68.2% nt identity with that of Calla lily chlorotic spot virus (CCSV), the highest among all compared viruses. This RNA was predicted to encode a non-structural protein (NSs) (459 aa, 51.7 kDa) and a nucleocapsid protein (N) (278 aa, 30.3 kDa). The N protein shared 85.8% amino acid identity with that of CCSV. The NSs protein shared 82.7% amino acid identity with that of Tomato zonate spot virus(TZSV). Conclusion: Our results indicate that the isolate 2009-GZT is a new species of Tospovirus, which is named Tomato necrotic spot virus (TNSV). This finding suggests that a detailed survey in China is warranted to further understand the occurrence and distribution of tospoviruses.

Objective Probe into the influence and the mechanisms of CCK and gastrin on the apoptosis of bile duct carcinoma cells.

Various extracellular matrix components were employed as coating materials to promote hepatic differentiation from ADSCs. However, no consensus was achieved about the optimal coating matrix due to the lack of direct comparison among different coating matrix. In this study, several coating extracellular matrixs were used for hepatic differentiation of ADSCs and direct comparison between them was performed. We demonstrated that liver DCM as coating matrix could significantly enhance the hepatic differentiation from ADSCs compared with collagen, fibronectin and Matrigel both in the presence and absence of GFs, including enhanced hepatocyte-specific genes expression, hepatocyte related protein secretion with improved liver functions. And the differentiated cells also exhibited the characteristics of mature hepatocytes. In conclusion, the study proved an effective hepatic-inducing method and indicated that DCM could promote the differentiation of ADSCs into hepatocyte-like cells, which demonstrates feasibility of liver DCM as a bio-scaffold for liver regenerative medicine and tissue engineering.

Cholecystectomy is routinely performed at most transplant centers during living donor liver transplantation (LDLT). This study was performed to evaluate the feasibility of liver graft procurement with donor gallbladder preservation in LDLT.Eighty-nine LDLTs (from June 2006 to Dec 2012) were retrospectively analyzed at our hospital. The surgical approach for liver graft procurement with donor gallbladder preservation was assessed, and the anatomy of the cystic artery, the morphology and contractibility of the preserved gallbladder, postoperative symptoms, and vascular and biliary complications were compared among donors with or without gallbladder preservation.Twenty-eight donors (15 right and 13 left-liver grafts) successfully underwent liver graft procurement with gallbladder preservation. Among the 15 right lobectomy donors, for 12 cases (80.0 %) the cystic artery originated from right hepatic artery. From the left hepatic artery and proper hepatic artery accounted for 6.7 % (1/15), respectively. Postoperative symptoms among these 28 donors were slight, although donors with cholecystectomy often complained of fatty food aversion, dyspepsia, and diarrhea during an average follow-up of 58.6 (44-78) months. The morphology and contractibility of the preserved gallbladders were comparable with normal status; the rate of contraction was 53.8 and 76.7 %, respectively, 30 and 60 min after ingestion of a fatty meal. Biliary and vascular complications among donors and recipients, irrespective of gallbladder preservation, were not significantly different.These data suggest that for donors compliant with anatomical requirements, liver graft procurement with gallbladder preservation for the donor is feasible and safe. The preserved gallbladder was assessed as functioning well and postoperative symptoms as a result of cholecystectomy were significantly reduced during long-term follow-up.