Objective: To compare post-orthotopic liver transplantation(OLT) survival rates between patients with recurrent HCC after partial hepatectomy and those with de novo OLT for HCC, and to screen for the risk factors associated with post-OLT mortality. Methods: From July 2003 to August 2005, 77 consecutive HCC patients underwent OLT, including 15 patients with recurrent HCC after partial hepatectomy for tumor resection (the rescue OLT group) and 62 patients with de novo OLT for HCC (the de novo OLT group); the post-operation survival rates were compared between the 2 groups. Thirty-three demographic, clinical, histological, laboratory and intra- and post-operative variables were analyzed. Cox proportional hazards regression model were used to screen for the factors associated with the survival rate. Results: The median age of the patients was 48.6 years and the median follow-up period was 20 months. Three patients(20.0%) in the rescue OLT group and 15 patients (24.2%) in the de novo OLT group died during follow-up(P=0.73). The 30 day-mortality of OLT was 6.7% for the rescue OLT group vs. 1.6% for the de novo OLT group(P=0.27). Cox proportional hazards model showed that the presence of pre-OLT hyperbilirubinemia, requirement of post-OLT transfusion, size of the largest tumor, tumor macroembolism and family history of HCC were significantly associated with a higher hazard for mortality. Conclusion: No significant difference is found in the survival rates between OLT as de novo therapy and OLT as a rescue therapy for patients with HCC. Pre-OLT hyperbilirubinemia, post-OLT requirement of transfusion, size of the largest tumor, tumor macroembolism, and family history of HCC are associated with a poor survival outcome.

Objective To study the prevention and treatment of chylous leakage after Whipple operation.Methods Clinical data of 381 patients underwent Whipple operation from Jan 2010 to march 2013 were retrospectively analyzed.Results Among 381 patients,23 patients had chylous leakage,the incidence was 6.04％.While in 89 patients,in which intraoperative precautionary physical ligation or mattress suturing of the lymphatic vessels were undertaken,no chyle leakage occurred ; Most of chyle leakage occurred in malignant tumor after radical resection(21 patients),most chylous leakage was found 5-8 days after surgery,with daily volume of 260-450 ml.All patients of chylous leakage were cured by conservative treatment.Conclusions Chylous leakage are common in PD.Intraoperative preventive measures such as lymphatic duct ligation or safe suturing can effectively prevent chyle leakage.Conservative therapy heals most chylous leakage after Whipple operation.

Objective To study the platelet antibody screening and crossing match by surface plasmon resonance(SPR) ,and to find a new way for platelet compatibility testing .Methods The corresponding universal platelet antigen was fixed on the SPR chip surface by the amino coupling method .Platelet antibody positive and negative control serum were analysed by SPR micro-ar-ray ,the stability ,sensitivity and specificity of this technique were discussed ,and compared with MAIPA assay .Finally we used the SPR technology to cross match for ten cases of the platelet antibody positive patients before infusion ,and to evaluate the effect of platelet infusion .Results For the SPR technology ,the stability ,sensitivity and specificity of platelet antibody detection were all better ,106 cases of the repeated platelet transfusion samples were tested by SPR assay and MAIPA method ,there was no signifi-cant difference between them(chi-square=0 .333 ,P>0 .05) ,the total consistency was 97 .2% .The 10 cases of platelet antibody positive patients were crossed match before platelet transfusion by SPR technology ,the good results of 8 cases of them were found by the clinical tracking evaluation ,1 h CCI>7 .5 ,24 h CCI>4 .5 .Conclusion SPR technology for screening platelet antibody mat-ches with MAIPA method in basic quality ,but SPR assay is simple ,rapid ,reliable and intuitive ,label free ,which can satisfy the re-quirements for clinical rapid detection of platelet antibody screening and crossing match .

Objective To investigate the application value of double-layer continuous suture in pancreatic jejunum anastomosis of pancreatoduodenectomy (PD).Methods A retrospective analysis of 114 consecutive patients (67 men and 47 women) who underwent PD from June 2012 to July 2013 were conducted. There were 79 patients who were treated by double-layer continuous suture technique and 35 patients by double-layer interrupted suture technique .The incidence of pancreatic fistula and other complications after PD with two different suture techniques were compared .Results The operation time of double-layer continuous suture group and double-layer interrupted suture group is respectively ( 284 ±5 ) and ( 288 ±7 ) mins, the intraoperative bleeding volume is respectively (236 ±29) and (282 ±49) ml, the differences between two groups were no statistical significance .The postoperative fasting time in two group was respectively (7.8 ± 0.5) and (9.7 ±0.5) days, the length of hospital stay time was respectively (14.0 ±1.0) and (17.2 ± 10.0) days, the incidence of postoperative pancreatic fistula ( POPF) was respectively 17.1%(6/35) and 39.2%(317/9 ), the differences between two groups were statistically significant (all P<0.05).Grade A POPF was found in 4 patients ( 11.4%) from the double-layer continuous suture group and in 5 patients (6.3%) from the double-layer interrupted suture group.Grade B POPF was identified only in 1 patients (2.9%) from the double-layer continuous suture group and in 23 patients (29.1%) from the double-layer interrupted suture group .The presence of Grade C POPF was only documented in 1 patient from the double-layer continuous suture group and in 3 patients from the interrupted suture group .Conclusions Continuous suture can be safely used in the duct-to-mucosa pancreatojejunostomy .The double-layer continuous suture can be more effective in reducing pancreatic fistula , improving the feeding time , and reducing the length of hospital stay, and it is worthy of clinical popularization and application .

Objective To study the changes of the peripheral blood regulatory B cells and related cytokines in children patients with chronic idiopathic thrombocytopenic purpura (CITP),and to analyze their correlations with effective Th cells,and to discuss their roles in the pathogenesis of CITP.Methods The peripheral blood mononuclear cells (PBMC)were separated in 30 cases of CITP,then percentages of Th1 ,Th17,Th22 and Breg cells were detected by the flow cytometry(FCM).IFN-γ,IL-17,IL-22 and IL-10 levels in culture supernatant were measured by the ELISA method.Results Compared with the control group,the proportions of the peripheral blood Th1 ,Th17,Th22 subsets in CITP children were increased[(18.4±4.7)% vs.(10.82±4.4)%;(2.42±1.02 )% vs.(1 .23±0.42)%;(0.79±0.24)% vs.(0.26±0.11)%],the proportion of Breg cells was reduced significantly,the differ-ences had statistical significance[(0.83±0.21)% vs.(1 .89±0.12)%,P <0.05].In CITP children,the IFN-γ,IL-17,IL-22 levels in culture supernatant were higher than those in the control group[(278.2±121.2)pg/mL vs.(181.8 ±82.2)pg/mL;(214.8 ± 100.5)pg/mL vs.(161 .4±67.8)pg/mL;(122.16±22.2)pg/mL vs.(90.8±31.1)pg/mL],but the IL-10 level was lower than that in the healthy control group[(27.4±12.6)pg/mL vs.(46.1±16.2)pg/mL),differences between them had statistical signifi-cance(P <0.05).Besides,there was a positive correlation between the proportion of Breg cells and cytokine IL-10 level(r=0.617, P <0.05 ),however,there was a negative correlation between the proportion of Breg cells with Th1 ,Th17 and Th22 cells (P <0.05),between IL-10 level with IFN-γ,IL-17 and IL-22 levels(P <0.05).Conclusion The downregulation of Breg cells proportion may participate in the immune disorder mechanism of effective Th cells in CITP children,which can provide a new idea for the im-munoregulation therapy in CITP children.

OBJECTIVETo assess the long-term clinical effectiveness of Chinese herbal medicines for benefiting qi and activating blood circulation (CHMBQABC) plus routine Western medical intervention in treating unstable angina (UA) patients of qi deficiency blood stasis syndrome (QDBSS) after percutaneous coronary intervention (PCI) based on Markov model.

METHODSA Markov model was established based on prognosis and sequelae of UA patients after PCI treated by CHMBQABC plus routine Western medical intervention or by routine Western medical intervention. According to the transition probabilities of 40 Markov cycles and quality-adjusted life years (QALYs) averagely gained, we assessed the therapeutic advantage of CHMBQABC plus routine Western medical intervention.

RESULTSBy the prediction of Markov model for 20 years, the transition probabilities of revascularization, non-fatal myocardial infarction, non-fatal stroke, and all-cause death in the CHMBQABC plus routine Western medical intervention group was 56.65%, 6.53%, 5.16%, and 31.66%, respectively, and the QALYs averagely gained was 12.95; while the transition probabilities of revascularization, non-fatal myocardial infarction, non-fatal stroke, and all-cause death in the Western medical intervention group was 55.31%, 6.87%, 5.25%, and 32.57%, respectively, and the QALYs averagely gained was 12.84. Compared with the Western medical intervention group, the QALYs averagely gained was 0.11 in the CHMBQABC plus routine Western medical intervention group.

CONCLUSIONBased on predicted results of the Markov model, CHMBQABC plus routine Western medical intervention got better efficacy in treating UA patients after PCI, indicating CHMBQABC plus routine Western medical intervention could improve the long-term clinical effectiveness for UA patients of QDBSS after PCI.

Aged ; Angina, Unstable ; drug therapy ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Markov Chains ; Middle Aged ; Models, Theoretical ; Phytotherapy ; Treatment Outcome

Ataxia Telangiectasia Mutated Proteins ; Breast ; metabolism ; pathology ; Breast Neoplasms ; metabolism ; pathology ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; metabolism ; pathology ; Cell Cycle Proteins ; metabolism ; Checkpoint Kinase 2 ; DNA-Binding Proteins ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Neoplasm Grading ; Protein-Serine-Threonine Kinases ; metabolism ; Tumor Burden ; Tumor Suppressor Protein p53 ; metabolism ; Tumor Suppressor Proteins ; metabolism

The gene of beta-glucuronidase from Thermotoga maritima was cloned into the plasmid pHsh, and expressed in Escherichia coli JM109. The recombinant protein was purified to homogeneity by a simple step, heat treatment. The recombinant enzyme had a molecular mass of 65.9 kD. The optimal activity of beta-glucuronidase was found at pH 5.0 and 80 degrees C. The purified enzyme was stable over a pH range from 5.8 to 8.2 and had a half life of 2 h at 80 degrees C. The kinetic experiments at 80 degrees C with p-nitrophenyl-beta- glucuronide as substrate gave a K(m) and V(max) of 0.18 mmol/L and 312 u per mg of protein. The purified enzyme could transform glycyrrhizin to glycyrrhetinic acid.
Cloning, Molecular ; Enzyme Stability ; Escherichia coli ; enzymology ; genetics ; Glucuronidase ; biosynthesis ; genetics ; metabolism ; Glycyrrhetinic Acid ; metabolism ; Glycyrrhizic Acid ; metabolism ; Hot Temperature ; Kinetics ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Thermotoga maritima ; enzymology ; genetics

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective:To detect the expression levels of neuropilin1 (NRP1)mRNA and miR-9 in non-small cell lung cancer (NSCLC)tissue samples, and to explore the correlations between the expressions of NRP1 mRNA, miR-9 and the clinicopathological characteristics of the patients with NSCLC.Methods:Informed consent was obtained from each patient before surgery.The tissue samples including 45 NSCLC tissue ,45 adjacent carcinoma tissue and 45 normal lung tissue were collected from China-Japan Union Hospital of Jilin University from 2010 to 2011.qRT-PCR was used to detect the expression levels of NRP1 mRNA and miR-9 in three kinds of lung tissue, and the correlation between the expressions of NRP1 mRNA, miR-9 and clinicopathological characteristics of the patients with NSCLC was analyzed.Results:Compared with normal tissue,the expression level of NRP1 mRNA in adjacent carcinoma tissue had no change (P>0.05),but the expression level of NRP1 mRNA in non-small cell lung cancer tissue was significantly decreased (P<0.05).Compared with normal tissue,the expression level of miR-9 in adjacent carcinoma tissue had no change (P>0.05),but the expression level of miR-9 in non-small cell lung cancer tissue was significantly increased (P < 0.05 ). Furthermore, in adjacent carcinoma tissue, the expression level of miR-9 in the males was lower than that in the females (P<0.05 ). In NSCLC tissue, the expression level of NRP1 mRNA had no relationship with sex,age,differentiation degree,TNM stage and clinical stage,but was significantly correlated to the histological subtype and lymph node metastasis (P<0.05).In NSCLC tissue,the expression level of miR-9 had no relationship with age, pathological type, lymph node metastasis, differentiation degree,TNM stage,and clinical stage (P>0.05),but was correlated to the sex (P<0.05). Conclusion:The expression level of miR-9 is up-regulated and the expression level of NRP1 mRNA is down-regulated significantly in non-small cell lung cancer tissue. The detection of the expression level of NRP1 mRNA contributes to j udge the histological subtype and lymph node metastasis of NSCLC.