Objective To explore the influence of abnormal donor hepatic artery on hepatic artery and biliary complications after liver transplanta?tion,and summarize the hepatic artery reconstruction procedures during transplantation. Methods The clinical data of 210 cases of liver transplan?tation conducted in our hospital from May 2005 to April 2015 were retrospectively searched for the study,including 42 with abnormal donor hepatic artery. Results Among the 210 liver transplantation,42 cases exhibited abnormal donor hepatic artery,and the aberration rate was 20.0%. Mean volume of blood flow of abnormal group and normal group was 4.7±95.1 mL/min and 190.9±101.6 mL/min,respectively. There was no statistic differ?ence(P=0.519). Twelve cases had arterial complications,the incidence rate was 5.71%,and there was no statistic difference between each group (χ2=0.72,P>0.05). Twenty five cases got biliary complications,the incidence rate was 11.9%,and there was no statistic differences between each group(χ2=0.05,P>0.05). Conclusion There was no statistic difference of mean volume of blood flow after arterial reconstruction between two groups. Liver transplantation with abnormal arterial reconstruction will not increase the incidence rate of arterial and biliary complications.

Objective To summarize experience for hepatic artery reconstruction in orthotopic liver transplantation(OLT).Method A retrospective analysis was made for 183 cases of orthotopic liver transplantation performed in our institute from May 1995 to december 2006.All the arterial reconstructions were performed with 6-0 polypropylene sutures in an interrupted fashion under a 3.5 magnification surgical loupe.Donor hepatic arteries were anastomosed at the origin of the celiac artery with a Carrel's patch or at the level of splenic artery confluence.Extra-anatomic arterial reconstruction was based on recipient aorta using donor iliac artery graft.OLT with routine anatomic arterial construction served as control.Heparin or low-molecule-weight heparin as a prophylactic anticoagulation therapy was maintained during and after operation if prothrombin time is less than eighteen seconds.Follow-up Doppler ultrasonography was used daily in the early postoperative period.Results Overall incidence of hepatic artery complications was 3.28%.Hepatic artery thrombosis(HAT)was observed in 5 cases.Hepatic artery stenosis(HAS)occurred in 1 patient.Routine anatomic arterial construction was performed in 89.07%(163)of cases,and HAT developed in 3(1.84%)cases.Extra-anatomic arterial reconstruction was carried out in 10.93%(20)of the patients;the presence of HAT was identified in 3(15.0%)cases(X2=9.73,P＜0.01).Thrombolysis,balloon angioplasty,and vascular stenting via hepatic artery were performed.One patient suffering from identified hepatic artery thrombosis died of liver failure 19 days post-op.The other 5 patients were cured successfully with patent blood flow by interventional therapy.Mortality related to hepatic artery complication was 16.7%.Conclusion HAT and HAS may be minimized by using gastroduodenal branch-patch anastomosis and postoperative anticoagulation.Close follow-up by Doppler ultrasonography helps to make a prompt diagnosis and reduce HAT-and HAS related graft loss.

Objective: To evaluate the targeting activity in the animal model with human hepatoma, the 131I-human antiHBsAg Fab radioimmunoimaging was explored. Methods: Radioimmunoimagings were taken on different intervals after injection of 131 I-human anti-HBsAg Fab to the nude mice and tissue distribution was measured. The human anti-HBsAg Fab was compared with the murine monoclonal antibodies. Results: The experimental group developed tumor positive images after 3 days of radio-labeled monoclonal antibodies injection, and the peak accumulation of radio-activity on the 5th day.Statistics indicated the tumor/liver ratio of the human anti-HBsAg Fab, murine monoclonal antibodies and the control groups were 5.4,4.0 and 0.9 respectively on the 7th day. Conclusions: Our results suggest that the 131 I-human anti-HB-sAg Fab has a considerable targeting activity, and provide an evidence that it can be used as a novel humanized carrer for targeting therapy of hepatoma.

ObjectiveTo investigate the methods of hepatic arterial reconstruction in orthotopic liver (transplantation)(OLT), and the prevention and management of associated complications, so as to improve the therapeatic results and donor survival rate of OLT.MethodsThe clinical data of 85 cases of orthotopic liver transplantation performed in our institute from May 1995 to May 2004 were retrospectively analysed. Hepatic artery reconstruction was made by use of Carrels patch of donor celiac artery or bifurcation of donor common hepatic artery and splenic artery anastoniosed to left and right hepatic artery of recipient in 16 cases((18.82)%), to bifurcation of gastroduodenal and proper hepatic arteries of recipient in 61 cases((71.76)%), or use of donor iliac artery interposition graft to abdominal aorta in 8 cases(9.42%). Regular heparin or low-molecular-weight heparin as a prophylactic anticoagulation therapy was maintained during and after (operation) as indicated by prothrombin time. Intra-and postoperative Doppler ultrasonography was used to (monitor) hepatic arterial blood supply.ResultsHepatic artery thrombosis(HAT) was observed in 1 case (intraoperatively)(overall incidence 1.2%).Thrombectomy and reconstruction of the arterial anastomosis were performed immediately.This patient currently has a survival time of 13 months without HAT. HAT was not observed in any of the other 84 cases during the follow up of 2 to 52 months.Conclusions Hepatic artery thrombosis may be minimized by proper selection of anastomotic site of hepatic artery reconstruction and (effective) use of postoperative anticoagulation.Close follow-up by Doppler ultrasonography may make a prompt diagnosis of HAT, salvage grafts and avoid re-transplantation.

Objective To investigate the biological effect of anti-leukemic cells induced by eosinophilic granulocyte (EOS) in bone marrow of patients with chronic myelogenous leukemia (CML).Methods The BCR-ABL fusion gene as well as the expression of IL-12 and IL-17 mRNA were performed by RT-PCR.The serum concentrations of cytokine IL-12 and IL-17 were determined by enzyme-linked immuno sorbent assay (ELISA).Immunochemistry staining and cytochemistry staining were used to observe the peroxydase (POX) and human Leukocyte antigen (HLA)-DR expression of EOS in bone marrow.Immunofluorescence staining was used to observe mannose receptor (MR),IL-12,IL-17A and IL-17receptor A (IL-17RA) expression of EOS.The results between the CML patients and the healthy controls were compared.Results Serum levels of IL-12 and IL-17 were higher in the 60 CML patients [(196.33 ±21.79) ng/L and (36.55-±3.01) ng/L] than those in the controls [(96.60 ±4.92) ng/L and (23.74 ±1.36) ng/L].In the 32 patients with activated EOS,the levels of IL-12 and IL-17 were (273.12 ± 17.16)ng/L and (40.11 ± 6.13) ng/L,which were significantly higher than those in the non-activated EOS [(126.16 ± 14.27) ng/L and (28.14 ±5.29) ng/L] (P values ＜0.01).IL-12 and IL-17 mRNA were expressed in activated EOS,while BCR-ABL fusion gene was not found.The amounts of EOS were increased abnormally in the bone marrow and peripheral blood of the CML patients with POX positive staining in the cytoplasm and weakly positive HLA-DR staining.It was observed easily by a microscope that EOS could attack leukemic cells in bone marrow through adhesion,capture and phagocytosis.Activated EOS could express IL-12,IL-17A and MR,which was related with the serum levels of these cytokines.Conclusions Activated EOS in bone marrow of CML patients could express IL-12 and IL-17.Activated EOS could induce coup injury to leukemic cell by releasing POX and expressing IL-12 and IL-17.It can also capture or swallow target cells via the expression of MR on the membrane.EOS may play an important role in the anti-tumor immunologic function in bone marrow of CML patients.

Objective Pancreas perfusion is an essential step in human islet isolation.To develop the new methods for introductal canulation,collagenase infusion and to observe their effects on islets isolation.Methods A total of 17 pancreases were digested from March 2005 to April 2010.The pancreases were distended by three methods:the standard method (n =3),the one-cannula method (n =11) and three-cannula method (n =3).In the standard group,the pancreases were completely cut into half at the mid-body.Two catheters were inserted into the main duct:one directed toward the tail and the other to the head.In the one-cannula method group,a long tube was inserted into the duct at the head,advancing to the tail In the three-cannula method group,pancreatic parenchyma was then minimally cut at the mid-body and three catheters were inserted into the main pancreatic duct:one at the head (the first catheter) and two at the mid-body,one toward the tail (the second catheter) and the other toward the head (the third catheter).The pancreases were digested by improved Ricordi technique.Ficoll continuous density/grads centrifuge method was performed to purify the islets.DTZ staining was adopted to identify islets and count islet equivalent (IEQ). AO/EB fluorescence examination was used to count active islet percentage.Static glucose stimulating test (SGS) in vitro was designed to estimate islet function and calculate SI.Results The distension volume of the threecannula method group was 1.24 rnl/g pancreas,and higher than the other groups (for the standard group:0.71 ml/g pancreas; for one-cannula method group:0.96 ml/g pancreas,P＜0.05).The yield of islet in the three-cannula method group and the one-cannula method group was 2514 and 2270 IEQ/g,which was significantly more than that in the standard group (1914 IEQ/g pancreas,P＜0.05).The purity and viability of the islets were 74 ％/79.3 ％,75.6 ％/79.4 ％ and 78.3 ％/84.0 ％ respectively in the three groups with the difference being not significant among the groups.SI in the one cannula method group (4.74) and the three-cannula method group (5.27) was significantly higher than that in the standard group (3.46).ConclusionThe three-cannula method improved collagenase infusion and the islet yields.

Objective To screen the mutation of KATP channel mutations in Chinese pedigrees with infantile onset type 1 diabetes mellitus (T1DM) and neonatal diabetes mellitus.Methods A cohort of 12 children of infant onset T1DM and neonatal diabetes mellitus admitted into Beijing Children's Hospital between March 2004 and June 2013 were selected.PCR amplification and direct sequencing were used to analyze the 39 exons of ABCC8 gene and one exon of KCNJ11.And the mutational sites of the parents of the probands was sequenced in order to identify the inheritance.Results Analysis revealed ABCC8 mutation in 25％ (3/12 cases) of the patients,a case of transient neonatal diabetes (TNDM),a case of permanent neonatal diabetes mellitus (PNDM) and a case of infant onset T1DM.All positive patients showed a known heterozygosis mutation in the ABCC8 gene(R1182Q,c.3545G ＞ A,D209E,c.627C ＞ G,E208K c.622G ＞ A).The residue R1182Q,which was located at a position involved in joining transmembrane domain 2 to nucleotide binding domain 2,the mutations E208K and D209E were located in the intracellular region that links the transmembrane domain with the gatekeeper module.All the three mutations were located throughout the cytoplasm part of SUR1 protein.The TNDM successfully transferred from insulin to oral sulfonylureas therapy.Conclusions There is a complex genetic pathogenesis in neonatal and infant-onset diabetes.The KATP channel activating mutations is one of the main causes of neonatal diabetes mellitus and may cause T1DM in infants in China.Oral Glibenclamide therapy seems highly effective for some patients with the KATP channel activating mutations.

Objective To analyze the clinical characteristics and gene mutations of 56 patients with congenital hyperinsulinism(CHI)and to provide a theoretical basis for clinical diagnosis and treatment of CHI.Methods Fifty-six children who were diagnosed as CHI between February 2002 and January 2016 in Beijing Children's Hospital Affiliated to Capital Medical University were selected as research subjects.A retrospective study was done about the clinical data and the treatment procedures of the 56 patients,such as perinatal conditions,clinical manifestations,laboratory data,treatments,prognosis and so on.Polymerase chain reaction(PCR)-DNA technology or next-generation sequencing technology was used to analyze the CHI relevant genes of the 56 patients.Results Thirty of the 56 patients carried CHI gene mutation.(1)Twenty-three of 56 patients(41.0％)carried ABCC8/KCNJ11 gene mutations:4 of 23 patients carried complex heterozygous mutation,1 of 23 patients carried both ABCC8 and KCNJ11 gene mutation,1 of 23 patients carried maternally inherited ABCC8 gene mutation,12 of 23 patients carried paternally inherited ABCC8 gene mutation,1 of 23 patients carried paternally inherited KCNJ11 gene mutation,3 of 23 patients carried de novo ABCC8 gene mutation,1 of 23 patients had unknown genetic way,19 of 23 patients were treated with Diazoxide,2 of 19 patients were responsive to Diazoxide,7 of 19 patients were unresponsive to Diazoxide and 10 of 19 patients were uncertain to Diazoxide.(2)Five of 56 patients(8.9％)carried GLUD1 gene mutation,4 of 5 patients were treated with Diazoxide and they were all responsive to Diazoxide.(3)One of 56 patients(1.7％)carried de novo GCK gene mutation,responsive to Diazoxide treatment.(4)One of 56 patients(1.7％)carried maternally inherited SLC16A1 gene mutation,responsive to Diazo-xide treatment.Conclusions The ABCC8 gene and GLUD1 gene mutation are the main causative genes of CHI.The GCK gene and SLC16A1 gene mutation are in the minority.Most ABCC8 gene and KCNJ11 gene mutation are unresponsive to Diazoxide treatment.

Objective:To study the effect of immunological molecules expressive state upon the telomerase activation ( TA) of bone marrow mononuclear cells ( BMMNC ) in the hemopoietic microenvironment of patients with immune related hematocytopenia ( IRHS) ,and to explore the immunologic mechanism as well as the clinical significance of hematoclasis in marrow of IRHS patients .Methods:①TRAP-PCR-ELISA method was performed to detect the TA of BMMNC in marrow of 366 IRHS patients before and after therapy.②The molecules HLA-DR,anti-hunman IgG,FcγⅡR,mannose receptor ( MR),IL-17A and its receptor ( IL-17AR) were analysed by immunochemistry and immunofluorescence staining .③The flow cytometric ( FCM) was used to analyse the proportion of CD3+CD4+T cells as well as CD3+CD8+T cells ,CD3-CD19+B cells and CD3-CD16/56+NK cells in peripheral blood lymphocyte.60 cases of health examination were selected as the control group , and 30 cases hypoferric anemia patients were selected as disease control.The differences between patient group and control group were analysed with statistic method .Immunochemistry and immunofluo-rescence staining were performed to in situ analyze the activation-characteristics of immunocyte in bone marrow slides of IRHS ,and the dependablity of cellular immunologic injury was also checked.Results: ①The levels of TA was 0.261 7 ±0.021 6 before treatment , higher than the disease control group (0.061 6±0.031 3 ,P<0.01).Among of them HLA-B27+patients were higher than HLA-B27-patients (0.301 3±0.020 6 vs.0.192 3±0.012 9,P<0.05).Serious IRP patients with HLA-B27+IgG+were obviously higher than HLA-B27-IgG+patients (0.401 6±0.017 2 vs.0.221 1±0.011 0,P<0.01).②In marrow of HLA-B27+IgG+patients,both cell immunity and humoral immunity were in disorder in the hemopoietic microenvironment ,and immonocyte in marrow expressed HLA-DR, FcγⅡR,IL-17A,IL-17RA and MR,and Th,Ts,B cell and NK cell in peripheral blood increased in different degree ,inducing the in-flammation of haemocyte and lead to destruction.③Humoral immunity was in the dominant level in morrow;humoral immunity of HLA-B27-IgG+patients,immonocyte expressed FcγⅡR in high level,but IL-17A was seldom expressed,only CD19+B cell was increased slightly ,the antibody dependent cellular cytotoxicity ( ADCC) was the main mode of destruction.After therapy glucocorticoids associated with ciclosporin A ,the TA level of BMMNC decreased to 0 with devitalization.Conclusion: The telomerase activation of bone marrow mononuclear cells in IRHS is related with the immune state of hemopoietic microenvironment and the pathologic lesion degree of hema -topoietic cell in marrow.It is viral infection and immunological activation as well as a variety of inflammatory factors play a part in the immunologic injury that might be an important factor of the enhancement in TA.

Objective To evaluate the effect of islet transplantation for patients with type 2diabetes mellitus (DM). Methods Since December 2007, 4 cases of islet transplantations were performed on 3 patients with type 2 DM and end-stage renal disease (ESRD). Two patients received simultaneous islet-kidney transplant from single-donor (SIK), and one received 2 consecutive islet transplants 5 months following kidney transplantion (IAK). All recipients given insulin with a dose of percutaneous transhepatic portal catheterization. Anti-CD25 monoclonal antibody was used as induction. For SIK, low-doses of Tacrolimus and sirolimus were used as maintenance immunosuppression protocol. For IAK, the maintenance protocol included cyclosporine and MMF.Insulin dose, the level of blood glucose, C-peptide and the value of HbA1 were observed. Results The first patient of SIKhad normal glucose level 3 days after surgery and became insulin independent within the first month, but insulin was administered gradually and the dose reduced to 1/3. The second patient of SIK died of bleeding and secondary infection of liver puncture site 5 days following operation, the blood glucose level recovered to normal 24 h after operation. The insulin dose of the patient of IAK was reduced to 1/2 after the first transplant. The patient became insulin free after the second operation. The level of fasting and postprandial C-peptide of the surviving recipients increased by 600 pmol/L. The value of HbA1 of the SIK was 6.7 %～7.3 %, while that of the IAK was 5. 5 %～ 5. 9 %. Conclusion Islet transplantation is an effective treatment for patients with type 2 DM.