Objective To explore the feasibility of laparoscopic cholecystectomy(LC) in patients over 85 years old.Methods Three-port LC was performed in 79 patients aged over 85 years old from October 1996 to October 2006.Patients' co-morbidities were effectively controlled before operation.The operation was conducted under continuous epidural anesthesia,and the CO2 pneumoperitoneum pressure was set at 8～10 mm Hg.Results The LC was successfully completed in 70 patients,while conversions to open surgery were required in 9 patients(11.4%).Causes of conversions included severe intraperitoneal adhesion in 6 patients,uncontrolled bleeding in 1 patient,and suspicion of gallbladder carcinoma in 2 patients.Postoperative complications occurred in 10 patients(12.7%,10/79).Of them,1 patient died of acute myocardial infarction,and the remaining 9 patients were cured(including left heart failure in 1 patient,pulmonary infection in 2 patients,ileus in 1 patient,wound haematoma in 1 patient,fungal enteritis in 1 patient,and effusion in the gallbladder bed in 3 patients).Conclusions Proper peri-operative management,effective control of co-morbidities,and careful surgical performance are essential for a safe LC in patients over the age of 85.

Objective To investigate the feasibility of spleen-and splenic vessels-preserving laparoscopic distal pancreatectomy for the treatment of pancreatic cystic tumor of body and tail.Methods The clinical data of a female patient with pancreatic cystic tumor of body and tail who was admitted to the Sun Yat-Sen Memorial Hospital of the Sun Yat-Sen University in March 2013 were retrospectively analyzed.Spleen-and splenic vesselspreserving laparoscopic distal pancreatectomy was determined as the optimal therapeutic method according to the physical examination and the results of computered tomography scan.Laparoscopic or open operation combined with distal pancreatectomy and splenectomy would be carried out as a candidate choice once it is hard to separate the splenic artery and vein from distal pancreas or to control the serious vessels hemorrhage.The patient was followed up by outpatient examination every 1 to 3 months up to March 2015.Results Spleen-and splenic vessels-preserving laparoscopic distal pancreatectomy was finished successfully.The operation time and volume of intraoperative blood loss were 192 minutes and 50 mL,respectively.The patient took out-of-bed for activity at postoperative day 1 without complications.The multiple severe microcystic pancreatic adenoma was confirmed by postoperative pathological examination,with a maximum diameter of 3.5cm.The leakage tube was removed at postoperative day 5.The levels of serum amylase at postoperative day 1,3,5 were normal.The patient was discharged at postoperative day 8 and got regular follow-up without bleeding,pancreatic fistula,infection and a symptom of epigastric pain or discomfort.Conclusion Spleen-and splenic vessels-preserving laparoscopic distal pancreatectomy has advantages of less traumas,faster postoperative recovery and a preservation of normal splenic function,deserving clinical application.

Objective To assess the effects of limited hepatectomy on liver regeneration and function recovery of cirrhotic animal model.Methods Cirrhotic rat models were first prepared by intraperitoneal injection of CCL4.After that,the cirrhotic animal models underwent 20％ hepatectomy (n =30).The cirrhotic animals that underwent sham operation (n =30) and normal animals that underwent 20％ hepatectomy (n =30) were used as control groups.From the time when cirrhotic models were prepared to 3 months after 20％ hepatectomy,the hepatic function,coagulation function were tested regularly.Western blotting and real-time PCR were carried out to test the protein and gene expression of TGF-β,HGF and PCNA.Results Hepatic fibrosis and cirrhosis were observed during the preparation of cirrhotic rat models by intraperitoneal injection of CCL4.The hepatic function and coagulation function of cirrhotic models were partly recovered 3 month after 20％ hepatectomy.The gene and protein expression levels of TGF-β in the liver of animal model get higher during the preparation of cirrhotic rat models.However,the gene and protein expression levels of TGF-β get lower in cirrhotic model undergoing 20％ hepatectomy,when compared with that in cirrhotic animals (sham operation) and normal animals (20％ hepatectomy).Furthermore,the gene and protein expression levels of HGF and PCNA get higher in cirrhotic model undergoing 20％ hepatectomy,when compared with that in cirrhotic animals (sham operation) and normal animals (20％ hepatectomy).Conclusions Experimental limited hepatectomy facilitates the liver regeneration and function recovery of cirrhotic animal model,which may provide a novel method for the prevention and treatment of cirrhosis using limited hepatectomy technique.

BACKGROUND: Recently, little attention has been paid to how to induce and identify the functions of differentiated cells in the methods for embryonic stem (ES) cells differentiation into hepatocytes. Whether the differentiated cells express functional characteristics of hepatocytes should be one of the markers to identify the hepatic differentiation of ES cells.OBJECTIVE: To direct mouse embryonic stem cells in vitro differentiation into functional hepatocytes by introduction of murine cholestatic serum in hepatocyte growth factor (HGF)-induced system.DESIGN: A controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery, the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: The experiment was carried out in the Medical Research Center of the Second Affiliated Hospital of Sun Yat-sen University from October 2004 to February 2007. The mouse E14 ES cell line was kindly provided by the Stem Cell and Tissue Engineering Center of Sun Yat-sen University. Twenty male SD rats, aged 2 weeks, were purchased from the Experimental Animal Center of Sun Yat-sen University. All animal experimental procedures were abided by the rules of animal ethnics.METHODS: The SD rats were undergone common bile duct ligation to induce cholestasis. Ten days after the operation, the whole blood of rats was collected to prepare cholestatic serum. The ES cells were cultured using hanging-drop method for 5-7 days to develop embryonic bodies (EBs). The dissociated EBs cells were then induced hepatic differentiation with spontaneous system, HGF (20 μg/L) system and cholestatic serum (5%) plus HGF (20 μg/L) system, respectively.MAIN OUTCOME MEASURES: The cellular morphologic changes were observed using transverse microscopy dynamically. (2) The cell staining for albumin, α-fetoprotein, CK18/19, glycogen, indocyanine green (ICG) and fluorescein diacetate (FDA) was done after 4 weeks differentiation. (3) The hepatocyte-specific metabolic functions of synthesizing albumin, triacylglycerol and urea nitrogen were assayed at 3 days interval.RESULTS: (1) The differentiation of ES cells cultured in spontaneous system was uncontrolled and the cells could grow into a wide range of three-germ cells. The HGF could promote ES cells differentiation into endoderm and mesoderm (myocardium). But the differentiated cells only expressed low levels of hepatic specific functions in these two induced systems. (2) Under cholestatic serum plus HGF system, the ES cells could differentiate into polygonal cells with very uniform morphology which were positive in glycogen, ICG and FDA staining and showed higher capabilities of synthesizing albumin, triacylglycerol and urea nitrogen than the differentiated cells in the other systems (P＜0.05-0.01).CONCLUSION: The cholestatic serum, a mimic pathological microenvironment in vitro, could effectively promote ES cells-derived hepatocytes induced by HGF to express high level of liver-specific metabolism functions.

AIM: To explore mechanism by which sodium butyrate induces mouse embryonic stem cells(ES) to differentiate into hepatocytes in vitro.METHODS: E14 mouse ES cells were cultivated in a routine way,and then cultivated in suspension to form embryonic bodies(EBs).EBs were transferred into 6-well culture dishes and 3 mmol/L sodium butyrate was added into the culture medium.Morphological changes were investigated by phase contrast microscopy.?-fetoprotein(AFP),albumin(ALB) and cytokeratin 18(CK18) were examined by immunofluorescence staining.AFP,ALB,?_1-antitrypsin(AAT) and TTR mRNA were assayed by RT-PCR.Proportion of ALB positive cells was analyzed by flow cytometry.Periodic acid Schiff(PAS) reaction and indocyanine green(ICG) uptake assay were performed to assess the characteristic hepatocyte function of the differentiated cells.RESULTS: In the presence of sodium butyrate,parts of ES cells differentiated into a population with epithelial morphology similar to mouse hepatocytes.AFP and TTR mRNA expression were observed at 7 d,and ALB and AAT mRNA expressed at 14 d.Hepatocytes specific markers,ALB,AFP and CK18 were positive expression in immunofluorescence staining at 14 d.PAS reaction and ICG uptake were positive for the hepatocyte-like cells.CONCLUSION: Mouse ES cells can be induced into hepatocyte-like cells by sodium butyrate efficiently,and these ES cells-derived hepatocytes possess characteristic hepatocytic function.

BACKGROUND: The immature dendritic cell (imDC) can induce immunological tolerance and has widely application in the field of organ transplant. At present, the methods of inducing imDC are insufficient, so the new induction method is demanding.OBJECTIVE: To investigate the effect of sodium butyrate (SB) on the maturation and immunological function of human peripheral blood-derived imDC.DESIGN: Controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery in the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: Five samples of human peripheral blood were obtained from the healthy volunteers (aged 20-23 years) of Sun Yat-sen University, totally 500 mL. Then peripheral blood mononuclear cells (PBMCs) and lymphocytes were isolated within 2 hours.METHODS: The experiment was carried out in the Medical Research Center of the Second Hospital Affiliated to Sun (1 mmol/L) was added for induction, while those supplemented with maturation promoting factor lipopolysaccharide (LPS)the beginning of induction, while LPS was added on the sixth day for second stimulation.MAIN OUTCOME MEASURES: Cell morphological change, flow cytometry was used to detect DC phenotype,FITC-labeled Dextran was used to detect the endocytosis of DC, the production of IL-12 was determined by means of enzyme-linked immunosorbent assay, and the proliferation of lymphocyte induced by DC was assayed with mixed lymphocyte reaction.expressions of CD80, CD83 and HLA-DR were significantly lower in the imDC of routine induction group following SB maturity promoting, compared with LPS group (P＜0.01). On the sixth day, LPS was added into the SB-induced imDC,Endocytosis of DC: The imDC of routine induction group possessed a significantly lower endocytic activity after induced by LPS, and there were extremely significant differences compared with blank control group and SB maturation Production of IL-12: The production of IL-12 in the mDC induced by LPS was significantly higher than that in control group, SB maturation promoting group and SB induction group, the mDC induced by LPS in routine induction group stimulated significantly stronger proliferation of lymphocyte (P＜0.01).

AIM: To investigate the immunological function of sodium butyrate-induced immature dendritic cells in vitro.METHODS: The human monocyte-derived dendritic cells were induced in the presence of human granulocyte macrophage-colony stimulating factor(GM-CSF) and interleukin-4 (IL-4), combined with sodium butyrate. The immunological function of sodium butyrate-induced dendritic cells was detected by the FCM, endocytic activity, T cells stimulatory proliferation capacity, and interleukin-12 (IL-12) production.RESULTS: Sodium butyrate could down-regulate the major histocompatibility complex(MHC) class II and costimulatory molecules of dendritic cells, increase the endocytic activity, induce a stage of T-cell anergey, and inhibit the T helper cell type 1-skewing factor IL-12 production. CONCLUSION: Sodium butyrate inhibits the maturation of dendritic cells and induces production of immature dendritic cells, which may help to explore the machenism of its epigenitic modification.

Objective:To study the expression of forkhead box P1 (FOXP1) in intrahepatic cholangiocarcinoma (ICC), and its clinicopathological and prognostic significance.Methods:The clinical data of ICC patients treated with radical resection at Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine from January 1, 2013 to December 12, 2019 were retrospectively analysed. Of 48 ICC patients, there were 24 males and 24 females, with age of (59.1±10.1) years old (range 42 to 83 years old). Their clinicopathological data, including age, gender, tumor size, degree of differentiation, and staging were recorded. Immunohistochemical method was used to detect the expression of FOXP1 protein in ICC cancer tissues and the corresponding adjacent normal tissues. Kaplan-Meier method was used to calculate survival rates and to construct survival curves of patients. Cox regression model was used to analyze factors affecting prognosis of patients.Results:Forty-eight ICC cancer tissues and 40 corresponding paracancerous tissues were collected. The positive rates of FOXP1 proteins in ICC were significantly lower than the adjacent normal tissues [54.2%(26/48) vs. 92.5%(37/40), χ 2=15.76, P<0.05]. The degrees of tumor differentiation, lymph node metastasis, organ invasion and TNM staging were related to expression of FOXP1 ( P<0.05). Forty-two patients were followed-up with a median follow-up time of 11.5 (7.75, 19.25) months. Cox multivariate analysis revealed that invasion to adjacent organs, lymph node metastasis, high TNM staging (stage Ⅲ) and negative expression of FOXP1 were independent risk factors affecting overall survival of ICC patients. The overall survival and recurrence-free survival of FOXP1-positive ICC patients were 17.5 months and 15.5 months, which were significantly higher than the 14.0 months and 11.1 months, respectively, in FOXP1-negative patients. Conclusion:Negative FOXP1 expression was closely correlated with aggressive biological behavior and poor prognosis of ICC. FOXP1 may be used as new diagnostic and therapeutic targets.

OBJECTIVETo evaluate the feasibility, safety and long-term outcomes of laparoscopic repeat hepatectomy for recurrent hepatocellular carcinoma (HCC) following previous resection.

METHODSBetween January 2003 and January 2011, 14 patients with recurrent HCC were carefully selected to undergo repeat laparoscopic hepatectomy, among which 9 patients were male, 5 patients were female, and the average age was 54 years. Prior to re-resection, all patients had undergone at least one open hepatectomy for HCC. The perioperative and long-term outcomes of these patients were retrospectively analyzed.

RESULTSRepeat laparoscopic hepatectomy for these 14 patients were successfully performed without major perioperative complications. The mean operative time, intraoperative blood loss and hospital stay were (124 ± 82) minutes, (112 ± 43) ml and (7 ± 4) days, respectively. The mean follow-up period was 23 months (range 14 to 42 months). At the time of follow-up, 11 patients were still alive, among which 3 patients developed recurrent disease and 8 patients remained disease free. One patient died of liver dysfunction at 21 months, and another 2 patients died of tumor recurrence at 17, 31 months, respectively.

CONCLUSIONLaparoscopic surgery for recurrent HCC remains a viable option for repeat hepatectomy in selected patients who have undergone open hepatectomy.

Adult ; Aged ; Carcinoma, Hepatocellular ; surgery ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Humans ; Laparoscopy ; Laparotomy ; Liver Neoplasms ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; surgery ; Retrospective Studies