Objective To investigate the risk factors of pancreatic fistula after pancreaticoduodenectomy.Methods The clinical data of 310 patients who received pancreaticoduodenectomy at the Ruijin Hospital of Shanghai Jiaotong University from January 2005 to May 2013 were retrospectively analyzed.The risk factors associated with the interoperative pancreatic fistula were analyzed.The univariate and multivariate analysis were done using the Pearson chi-square test and non-conditional Logistic regression model.Results A total of 134 patients had postoperative complications,including 103 (33.23％) with pancreatic fistula,among them 40 patients developed additional complications.The results of univariate analysis showed that preoperative levels of hemoglobin,total bilirubin,diameter of the pancreatic duct and postoperative level of albumin were risk factors of pancreatic fistula after pancreaticoduodenectomy (x2 ＝ 4.543,6.087,6.265,5.311,P ＜ 0.05).The results of multivariate analysis showed that preoperative level of total bilirubin equal to or above 34.2 μmol/L,the diameter of the pancreatic duct under 3 mm and the level of postoperative albumin under 28 g/L were the independent risk factors of pancreatic fistula (OR =1.806,1.936,1.780; 95％ confidence interval:1.107-2.948,1.170-3.206,1.002-3.165,P ＜ 0.05).Conclusion Preoperative jaundice (the level of total bilirubin ≥ 34.2 umol/L),pancreatic duct diameter ＜ 3 mm and postoperative malnutrition (albumin ＜ 28 g/L) indicate a higher incidence of postoperative pancreatic fistula.

OBJECTIVE:To create an in vitro harvesting method of culturing a large number of adult bone marrow MSCs(BMSCs) DESIGN,TIME AND SETTlNG:The randomized, controlled study was performed at the Shanghai Institute of Digestive Surgery (Key Laboratory of Education Committee of Shanghai City),as well as Department of General Surgery and Organ Transplantation Center,Ruijin Hospital,Medical College.Shanghai Jiao Tong University from September 2005 to April 2006.MATERIALS:Bone marrow samples were collected from normal persons.who did bone marrow examination at the Department of Hematology,Ruijin Hospital,Medical College.Shanghai Jiao Tong University.Donors were volunteers who signed the informed consent.METHODS:Human BMSCs were harvested using Pemoll gradient centrifugation and adherence method.and then incubated in microcarrier cytodex3.Common monolayer polystyrene was incubated as controls.Cell phenotype and proliferative activity were tested utilizing flow cytometry and MTT.MAIN OUTCOME MEASURES:Collection.incubation,morphology of human BMSCs.and prolireration and cell cycle of human BMSCs on the cytodex 3 were measured.RESULlTS:Flow cytometry detection showed that the surface marker of human BMSCs on the cytodex3 was ldentical to that on the common monolayer polystyrene;BMSCs were positive for CD29,CD44 and CD105.but negative for CD14,CD34,CD45,VLA-1 and HLA-DR.MTT detection demonstrated that human BMSCs were in the adaptive phase at days 1-3.and entered logarithmic phase frOm day 3.No significant difference was detected in human BMSCs on the monolayer polystyrene and cytodex3(P＞0.05).On the monolayer polystyrene,human BMSCs entered degenerating stage from day 6,whereas on the cytodex3,human BMSCs were still in the logarithmic growth phase at day 9(P＜0.05).Flow cytometry detection confirmed that the cell cycle of human BMSCs was the same both on the monolayer polystyrene and cytodex3 (P＞0.05). CONCLUSION:Using cytodex3 culture technique,a large amount of human BMSCs can be obtained,and the proliferative activity of these BMSCs is good.

BACKGROUND: Hepatocyte/polymer interface with good biocompatibility is the key factor in bioreactor design andconstruction, however, bioreactor used in the clinical practice currently is not an ideal one.OBJECTIVE: To establish human hepatocyte compatible polypropylene interface and to lay a foundation for establishingbioartificial liver reactor with polypropylene hollow fiber.DESIGN, TIME AND SETTING: The comparative observation, cell compatibility experiment was performed betweenFebruary and October 2003 at Shanghai Jiao Tong University, Shanghai, China.MATERIALS: Polypropylene Photochemical graft polymerization modification technique was used to graft hydrophilicacrylamide groups on the surface of polypropylene membrane by chemical bonds to form modified polypropylenemembrane.METHODS: L02 human hepatoeytes were seeded on polypropylene membrane, modified polypropylene membrane andpolystyrene membrane, and polystyrene membrane was used as normal control.MAIN OUTCOME MEASURES: Static water contact angle of polypropylene membrane before and after graftmodification; morphology, adherent rate and proliferation activity of L02 human hepatocytes on different material surfaces.RESULTS: Static water contact angle after polypropylene membrane graft modification was smaller than that before graftmodification (P < 0.05). The adherent rate of L02 human hepatocytes on the surface of modified polypropylene membranewas 0, and the proliferation activity of them, which grew as spherical aggregates, was markedly higher than that of cells onpolystyrene membrane and polypropylene membrane without graft modification.CONCLUSION: Grafting polyacrylamide on the surface of polypropylene can establish good interface of L02 humanhepatocytes/polypropylene and form hepatocyte spherical aggregates through simple static culture.

Objective To identify the role of p53 in the induction of growth arrest DNA damage-inducible gene 45β (GADD45β) in HCC cells by Oxaliplatin.Methods A Hep3B+p53 clone was established by transfection of the full-length p53 sequence to Hep3B.Following oxaliplatin administration,quantitative real-time PCR was employed to validate the expression changes of GADD45β.pGL3 basic luciferase plasmids including promoter fragments were synthesized in vitro and transfected into cells.The effects on promoter activity,cell growth and the cleavage of Caspase-3 were further focused on.Results Hep3B+p53 expressed p53 protein stably.The transfection of p553 enhanced the induction of GADD45β in Hep3B by Oxaliplatin.The promoter activity of fragments constructed NF-κB and E2F-1 binding sites was induced about 1.5 and 0.8 folds by transfection of p53.The colony formation and DNA syntheses were inhibited apparently in Hep3B+p53 with p53 by Oxaliplatin (30.41％ and 75.60％ by 100 μmol/L Oxaliplatin,respectively).Moreover,p53 transfection triggered cleavage of Caspase-3 more rapidly.Conclusion p53 played a role in the induction of GADD45β in Hep3B by Oxaliplatin.

Objective To investigate the protective effect of polymyxin B (PMB) to the liver graft after liver transplantation and the underlying mechanism in rats.Methods Male SD rats were selected as the donors and recipients.Non-artery whole liver transplantation model was established in rats according to Kamada's two-cuff method.The rats were divided into two groups by the way of random number table method:control group (normal saline,0.5 ml) and PMB group (PMB,1 mg/ml,0.4 mg/kg+ normal saline 0.5 ml).The levels of portal vein plasma endtotoxin (EU/ml)were determined by endotoxin-analyzing machine of BET-24A. ALT,BUN,and TNF-α,IL-6 in serum were measured by using machine of Automatic Analyzer and ELISA,respectively.The CD14,TLR4,NFκB and AP-1 in the grafts were measured by RT-PCR and Western blotting,and pathological changes were observed. Results PMB decreased the levels of portal vein plasma endotoxin 1 h after reperfusion in PMB group as compared with control group (P＜0.05),and the levels of portal vein plasma endotoxin returned to the normal levels 6 h after reperfusion in both two groups (P＞0.05).After operation,the levels of ALT,TNFα and IL-6 in serum were significantly reduced (P＜0.05),the expression of CD14 and TLR4 mRNA in the grafts was significantly decreased (P＜0.05),the expression of Hsp60 protein and mRNA,and NF-κB and AP1 proteins in the grafts were reduced (P＜0.05),and the pathological damage to the grafts was significantly alleviated in PMB group as compared with control group.Conclusion PMB reduced the levels of portal vein plasma endotoxin after reperfusion in liver transplantation in rats.PMB improved liver function,reduced the injury of inflammatory response,decreased the levels of endotoxin signal pathway markers and alleviated the pathological damage to the grafts.

Objective To study the relationship between hepatic arterial buffer response (HABR),recovery of liver function,early biliary complications and small-for-size syndrome (SFSS).Methods Early hepatic hemodynamic parameters (including hepatic arterial flow (HAF),portal venous flow (PVF) were measured using duplex Doppler sonography in 34 patients who received living donor liver transplantation (preoperatively n＝26,intraoperatively n=26) and on postoperative days 1,2,3,and 7.Alanine aminotransferase (ALT),aspartate aminotransferase (AST) and total bilirubin (TBIL) level were measured preoperatively and on postoperative days 1,2,3,7,14,21,and 28.If TBIL level was elevated,we used B ultrasonography or CT and even ERCP to diagnose early biliary complications.The days taken for AST,AI T and TBIL to recover and the number of patients with early (＜60 days) biliary complications (bile leakage or bile stricture) and with small-for-size syndrome (SFSS) were recorded.Results Passive hepatic artery buffer response (HABR) was present in 11 patients early after living donor liver transplantation (group 1) and it disappeared in 23 patients (group 2).The recovery in days taken for normalization of AST (10.6± 8.8),AIT (11.6±9.0) and TBlL (average of 29) in group 1 were shorter than in group 2.However,the differences did not reach statistics difference (P＞0.05).The overall incidences of early biliary complications and small-for-size syndrome (SFSS) in group 1 were significantly lower than in group 2 (P=0.04).The survival rate in group 1 was 82 ％,compared with 74 ％ in group 2.Conclusions Passive hepatic arterial buffer response (HABR) disappeared in some patients early after living donor liver transplantation.There were high incidences of early biliary complications and small-for-size syndrome (SFSS) in these patients.Measurcment of hepatic buffer response in the early stage after living donor liver tranaplanta tion is valuable for predition of early biliary complications and small-for-size syndrome (SFSS),thus helping to prevent failure in transplantation.

Objective: To investigate the effect of global end-diastolic volume index (GEDI)-guided fluid resuscitation on the prognosis of patients with chronic heart failure and septic shock. Methods: This study was a prospective randomized controlled study. Consecutive eligible patients were divided into 2 groups according to the random number table method: control group (n=21) and experimental group (n=20). On the basis of routine treatment, patients in the control group received early goal-directed therapy until the central venous pressure (CVP) reaching 8-12 mmHg (1 mmHg=0.133 kPa), mean arterial pressure reaching over 65 mmHg, urine volume reaching over 0.5 ml·kg^-1·h^-1, and central venous oxygen saturation reaching more than 70%. On the basis of routine treatment, patients in the experimental group were monitored continuously on cardiac output with pulse indication and fluid resuscitation guided by volume index GEDI. The GEDI should be maintained on the range of 680-800 ml/m². The remaining resuscitation goals were the same as control group. General clinical data of the two groups were collected at admission. Negative fluid balance onset time, duration of mechanical ventilation, ICU mortality and 28-day mortality were compared between the two groups. The outcomes were recorded as listed: start time of negative fluid balance, duration of mechanical ventilation, mortality in ICU and 28-day mortality. Results: There was no significant difference in age, sex, weight, APACHE Ⅱ score, SOFA score and NYHA functional class score between the two groups (all P>0.05). The negative liquid balance onset time in the control group was 3.5 (2.5, 4.0) days, which was significantly longer than that in the experimental group (2.6 (2.0, 3.0) days,U=115.0, P=0.012). The duration of mechanical ventilation was 355 (118, 552) hours in the control group, which was significantly longer than that in the experimental group (132 (36.75, 233.3) hours, U=130, P=0.038). The ICU mortality was 38.1% (8/21) in the control group, tended to be higher than that in the experimental group (20.0%(4/20), χ²=1.620, P=0.203). The 28-day mortality was 42.9% (9/21) in the control group, similar as in the experimental group (25.0%(5/20), χ²=1.482,P=0.477). Conclusion Fluid resuscitation guided by volume index (GEDI) may improve the prognosis of patients with chronic heart failure complicated with septic shock.

The present study was to investigate in vitro the rat cardiomyocyte injury with targeting of home-made perfluorocarbon lipid particles with avidin-biotin interaction. Neonatal rat cardiomyocytes were cultured in vitro and divided into two groups: TNF-alpha activated group and non-activated group. Those in the TNF-alpha activated group were exposed to 200 ng/ml TNF-alpha solution for 6 hours and then cardiomyocytes in both groups were pretargeted with biotinylated ICAM-1 monoclonal antibodies, and were exposed to streptavidin, and then to homemade green fluorescently-labeled biotinylated perfluorocarbon lipid particles. Cardiomyocytes nucleus stained with Hoechst. The results were detected with fluorescence microscope. As a result, in TNF-alpha activated group, around blue fluorescent cardiomyocytes nucleus, a great amount of green fluorescent particles were found, while there were few green fluorescent particles in non-TNF activated group. It has been shown that ICAM-1 is expressed in the surface of cardiomyocytes when they are stimulated by TNF-alpha. Perfluorocarbon lipid particles associated with ICAM-1 monoclonal antibodies can be targeted to injured cardiomyocytes by avidin-biotin interaction.
Animals ; Animals, Newborn ; Antibodies, Monoclonal ; metabolism ; Cells, Cultured ; Contrast Media ; Female ; Fluorocarbons ; immunology ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Lipids ; chemistry ; Male ; Microspheres ; Myocytes, Cardiac ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; pharmacology ; Ultrasonography

Chylous fistula is a postoperative complication following pancreatic surgery,it is mainly diagnosed according to the nature of the patient's drainage fluid (≥3 days after surgery,triglyceride concentration ≥ 1.2 mmol/L).The mechanism of occurrence includes obstruction,injury and exudation of the lymph-vessels.Risk factors can be concluded into 3 aspects:clinicopathological features,surgical procedure and postoperative management.Most of the chylorrhea can be cured by conservative treatment like modified dietary measures and somatostatin,while severe cases still requires intervention and surgical treatment.This article reviewed the risk factors and treatment approaches of postoperative chylous fistula,and elucidated relative mechanisms,hoping to provide guidance in clinical prevention,diagnosis and treatment of chylous fistula.

Objective:To explore the guiding effect of peripheral perfusion index (PI) on fluid resuscitation in patients with septic shock.Methods:Sixty-five patients with septic shock who were diagnosed according to relevant criteria of septic shock and admitted to the department of critical care medicine of the Affiliated Hospital of Hangzhou Normal University from September 2017 to December 2020 were included. Patients were divided into the conventional treatment group (30 cases) and PI guidance group (35 cases) by random number method. Both groups of patients were treated with the bundle according to clinical guidelines. Sputum, urine and blood were collected for pathogenic microorganism culture before the application of antibiotics, and vasoactive drugs were given. Both groups need to achieve all the following resuscitation goals within 6 hours: urine output > 0.5 mL·kg -1·h -1, mean arterial pressure (MAP) ≥ 65 mmHg (1 mmHg ≈ 0.133 kPa), central venous pressure (CVP) was 8-12 mmHg, and central venous oxygen saturation (ScvO 2) ≥ 0.70. There was no further resuscitation in the conventional treatment group after the goals were achieved. In addition to these four goals, the PI guidance group was expected to achieve PI≥ 1.4. Heart rate (HR), CVP, MAP, ScvO 2, blood lactic acid (Lac), the time of fluid negative balance, intensive care unit (ICU) mortality and 28-day mortality between the two groups were compared before and after 6 hours of fluid resuscitation. Results:Before fluid resuscitation, there were no statistically significant differences in all indicators between two groups. After 6 hours fluid resuscitation, the four treatment goals in PI guidance group were slightly lower than those of the conventional treatment group [HR (times/min): 96.5±12.1 vs. 97.7±7.9, MAP (mmHg): 83.2±6.2 vs. 82.1±7.5, ScvO 2: 0.661±0.077 vs. 0.649±0.051, CVP (mmHg): 10.8±2.7 vs. 10.4±2.1], there were no statistically significant differences between the two groups (all P > 0.05); the Lac level of the PI guidance group after resuscitation was lower than that of the conventional treatment group, and the difference was statistically significant (mmol/L: 4.8±1.3 vs. 5.9±1.4, P < 0.05); the duration of fluid negative balance in the PI guidance group was earlier than that in the conventional treatment group [days: 3.0 (2.0, 3.0) vs. 3.5 (3.0, 4.0), P < 0.05]. The ICU mortality and 28-day mortality in the PI guidance group were lower than those in the conventional treatment group [ICU mortality rate: 37.1% (13/35) vs. 50.0% (15/30), 28-day mortality rate: 57.1% (20/35) vs. 60.0% (18/30)], but the differences were not statistically significant (both P > 0.05). Conclusions:The peripheral PI can be used as an important indicator of fluid resuscitation in patients with septic shock. PI guiding fluid resuscitation in patients with septic shock can reduce Lac levels, shorten the duration of fluid negative balance and reduce the risk of fluid overload.