Objective To investigate the application of laparoscope,duodenoscope and choledochoscope in the treatment of intra-and extrahepatic bile duct stone.Methods The clinical data of 3 16 patients with intraand extrahepatic bile duct stone who were admitted to the Bethune First Hospital from April 2007 to August 2010were retrospectively analyzed.There were 269 patients with cholecystolithiasis and choledocholithiasis,10 patients with cholesystolithiasis,choledocholithiasis and hepatolithiagis,and 37 patients with choledocholithiasis.Laparoscopic cholecystectomy(LC)+laparoscopic common bile duct exploration(LCBDE)+choledochoscopy was applied to patients with hepatolithiasis or with the diameter of common bile duct≥10 mm;endoscopic sphincterotomy (EST)+LC or LC+EST was applied to patients with the diameter of common bile duct between 10 mm and 5 mm and the diameter of cystic duct<5 mm;LC+laparoscopic transcystic common bile duct exploration(TC-CBDE)+choledochoscopy wag applied to patients with the diameter of common bile duct≤5 mm and the diameter of cystic duct≥5 mm.Results The success rate of operation was 96.8%(306/316).A total of 163 patients received LC +LCBDE+T-tube drainage+choledochoscopy,and the mean operation time,expense,duration of hospital stay were 93.6 minutes,2.8×104 yuan and 9.8 days,respectively,and 5 patients had complications postoperatively.Fifty-four patients received EST+LC,and the mean operation time,expense,duration of hospital stay were 45.0minutes,6.6 days,2.3×104yuan,respectively,and 1 patient had complication postoperatively.Sixty-seven patients received LC+EST,and the mean operation time,expense and duration of hospital stay were 40.0minutes,6.1 days,2.4×104 yuan,respectively,and 2 patients had complication postoperatively.Thirty-two patients received one-stage repair of common bile duct and LC+TC-CBDE+choledochoscopy,and the mean operation time,expense and duration of hospital stay were 97.3 minutes,7.3 days and 2.5×104yuan,respectively,and 1 patient had complication postoperatively.A total of 272 patients were followed up for 12 months,except for 6 patients with recurrence of common bile duct stone,no residual stone or biliary stricture was etected.Conclusion Combined application of laparoscope,duodenoscope and choledochoscope has advantages of less trauma,quick ecovery and fewer complications in the treatment of intra-and extrahepatic bile duct stone.

BACKGROUND: Multipotency of hepatic stem cells is of important value in liver transplantation. Stem cells have been successfully identified and isolated from the animal livers. However, reports on whether stem cells exist in human hepatic tissue and how to isolate and identify them ars few.OBJECTIVE: This study was in attempt to isolate hepatic stem cells from human para-cancerous tissues of hepatoma and in vitro culture them, also to identify the stem cell surface marker, in order to find a new source of heptatic stem cells.DESIGN: Cell observation experiment.SETTING: Department of Common Surgery, First Hospital, Jilin University; Department of Common Surgery,Dongfeng Hospital of Traditional Chinese Medicine.PARTICIPANTS: Samples were harvested from 10 patients with hepatoma admitted to Department of hepatobiliary surgery, First Clinical College, Jilin University between October 2005 and June 2006, with age of 45 to 58 years.Hepatic tissue 2 cm away from cancer nest was cut when patients underwent hepatectomy, and it was pathologically confirmed as carcinoma-free tissue. Written informed consents were obtained from each patient. DMEM/F12 dry powder used for cell culture was provided by Hyclone Company, USA. Fresh fetal bovine serum was prepared by Lianxing Biotech Co.,Ltd, Tianjin. Various cell growth factors were the products of Cytolay Company, USA.METHODS: Para-cancerous tissues of hepatoma was cut into pieces, rinsed with Hank's solution and digested with type Ⅳ collagenase. Then the isolated cells were re-suspended in the DMEM/F12 medium supplemented with 0.1 volume fraction of fetal bovine serum, and hepatocyte growth factors, epidermal growth factors and α- fibroblast growth factors of 25 μg/L each were added in the above medium. When the cultured cells covered 2/3 of bottom,they were digested with trypsinase for passage and inoculated at 2×107 L-1. When cells propagated to the 3rd and 4th generations, 2.60×109 L-1 cell suspension prepared with trypsinase was added, and subsequently, anti-human C-kit antibody, immunomagnetic beads and Buffer solution were added in order. C-kit+ cells were preliminarily isolated by immunomagnetic bead separation. Haematoxylin-eosin staining and immunofluorescent histochemical double-staining were used for detecting the hepatic stem cells in para-cancerous tissues.MAIN OUTCOME MEASURES: ① Observation of cell morphology. ② Identification of hepatic stem cells from para-cancerous tissues. ③ Identification of C-kit+ cells by immunofluorescent histochemical double-staining.RESULTS :① After primarily cultured for 2 weeks, the adherent cells grew in colony. After one half of culture medium was renewed, mature hepatocytes were gradually broken and disappeared. Small round cells propagated, and most of them were located in the center and arranged in cluster. Most cells were found with one big nucleus in each, less cytoplasm and clear cell boundary. When cells propagated to the 1st and 2nd generations, they still grew in colony, but fast. Each C-kit+ cell isolated by immunomagnetic bead separation presented a spherical cell body with a very big nucleus and less cytoplasm. After in vitro cultured for 1 week, it presented broken pieces and apoptotic symptoms.② After para-cancerous tissue was stained by haematoxylin-eosin, atypically proliferated biliary tracts with small round cells could be seen in the portal area. After para-cancerous tissue was stained by immunofluorescent histochemical double-staining, small round cells in the biliary tracts proliferated in the portal arsa co-expressed red fluorescence AFP and green fluorescence cytokeratin (CK) 19 with yellow superposition arsa. ③ After C-kit+ cells were stained by fluorescence immunocytochemisty, cytoplasm expressed alpha-fetoprotein (AFP) red granules and CK19 green granules. The superposition area of both presented yellow fluorescence of AFP+/CK19+-positive cells.CONCLUSION: Hepatic stem cells exist in human para-cancerous tissues of hepatoma. Therefore, expressions of C-kit+/AFP+/CK19+, the surface markers of hepatic stem cells, can be used for identifying and isolating hepatic stem cells. Small round cells obtained by in vitro isolation and culture, i.e. hepatic oval cells possess bipotential differentiation of hepatocyte and hepatobiliary epithelial cells.

Objective To explore the indications,surgical techniques and clinical effects of laparoscopic splenectomy on patients with portal hypertension. Methods Analysis the clinical data of 32 patients of portal hypertenision with laparoscopic splenectomy and open splenectomy from March 2006 until June 2009. Results The effectiveness of the procedures for portal hypertension was evaluated.Among 16 patients with LS, 2 patients were converted to open surgery. There is no significant difference with operatative time, blood loss and hospital fees between the two groups, the laparoscpy group had the shorter mean hospitalization, fasting and draining time. Conclusions If there are enough preparation preoperation, skillful laparoscopic technique and micromesh manipulation during operation,laparoscopic splenectomy is a minimally invasive and safe technique in the patients with lower-grade to medium varicose veins.

Objective To compare the effect of naive porto-azygous devascularization and that as a remedy therapy after a failed endoscopy for the treatment of bleeding portal hypertension.Method From June 2005 to June 2010,230 portal hypertension patients were treated with porto-azygous devascularization,among them,202 cases were of portal hypertensive cirrhotics,28 cases of alcoholic cirrhosis.Group A (16 patients)received remedial porto-azygous devascularization after endoscopic treatment failed (esophageal variceal ligation,esophageal variceal selerotherapy).Group B(214 patients)received naive porto-azygous devascularization.Results The average operation time in group A was 198 min(115-335)min,mean bleeding amount was 750 ml(300-2000)ml,average post-operative hospital stay was 11 days (8-15)days.The average operation time in group B was 120 min(90-190)min,mean blood loss was 250 ml(150-500)ml,average post-operative hospital stay was 7 days(6-9)days.The average operation time,bleeding amount and post-operative hospital stay was significantly different between the two groups (P＜ 0.05).212(92.2％)patients were followed up,and the mean follow-up time was 2.5 years (1-6)years,mortality rate was 4.2％,rebleeding rate was 5.7％.Conclusions Surrounding the perioesophageal and gastric tissues were fibrotic and hard after the endoscopic treatment,this significantly increased the difficulty of surgery,therefore,porto-azygous devascularization is the choice of therapy for portal hypertension patients.

Objective To evaluate the safety and feasibility of total laparoscopic hepatectomy for liver neoplasms.Methods Laparoscopic hepatectomy in 15 patients with liver neoplasms were completed by combined application of ultrasound scalpel,Ligasure and vascular clip without blockage of liver blood flow,including 9 cases of hepatic cavernous hemangioma,whose diameters were from 5.0 cm to 15.0 cm,3 cases of hepatic cyst with fibrosis,located in left lateral hepatic lobe,3 cases of primary hepatic carcinoma,whose diameters were from 1.0 cm to 5.0 cm and the hepatic functions were all Child A.Results Laparoscopic hepatectomy was completed successfully in all 15 cases with no conversion to open laparotomy,including 6 cases of left lateral hepatectomy,9 cases of irregular hepatectomy.The mean operative time was 110 min,blood loss during operation was from 30 ml to 500 ml,the average was 251 ml.The mean postoperative hospital stay was 6.5 d.The mortality rate was 0%.No severe complications occurred except 1 case of small amount bleeding which stopped itself.Conclusion Total laparoscopic hepatectomy is a feasible,safe and minimal invasive approach for patients with liver neoplasms within segment Ⅱ、Ⅲ、Ⅳa、Ⅴ、 and Ⅵ.

Objective To summarize the clinical applications and surgical technique of laparoscopic distal pancreatectomy (LDP). Method The clinical data of 10 cases of pancreatic body and tail tumors undergoing laparoscopic distal pancreatectomy were retrospectively analyzed.Results Laparoscopic distal pancreatectomy (LDP) was successfully undertaken in 8 cases (including spleen preserving distal pancreatectomy in one case ). Intraoperatively two cases were converted to open surgery because of peripancreatic organs involvement by cancer in one case and massive bleeding in another case during laparoscopic procedures. The average operation time of LDP was 141 ± 35 min (95 -195 min),mean blood loss was 263 ± 151 ml( 100 -600 ml), average postoperative hospital stay was 7 ± 1 days (5 -9 days ). There was no major postoperative complications and no mortality. Final pathology was solid psedopapillary tumor in 4 cases, mucinous cystadenoma in 3 cases and islet cell tumor in 1 case, pancreatic ductal adenocarcinoma in 2 cases, hence 80% of tumors were benign. Conclusions LDP is indicated for benign body and tail pancreatic tumors and early malignant tumor of pancreatic body and tail. Being less traumatic, and fewer complications, LDP is a safe, effective and minimally invasive therapy.

Objectiye To investigate the clinical application value of emergency therapeutic encoscopic retrograde cholangio-pancreatography (ERCP) on acute biliary-pancreatic diseases.Methods Duodenoscope was applied on the emergency treatment for 103 patients of acute biliary-pancreatic diseases from March 2007 to May 2010. According to patients' conditions, the examination of ERCP,endoscopic nasobiliary drainage (ENBD) and endoscopic sphincterotomy (EST) were carried out. The papilla sphincter was slivered by needle knife under the circumstances of the stone incarceration in the papilla,then calculus was removed by net basket. ENBD or endoscopic retrograde biliary drainage (ERBD)was placed according to the state of an illness. Statistical analysis was made by SPSS 13.0 software with a 2 × 2 table analysis, Comparison of ratio were determined with x2 test, a value less than 0. 05 was considered as significant. Results Of 103 cases, the success rate of endoscopic treatment was 97. 1% ( 100/103 ).Among 100 cases, 25cases (24. 3% ) were treated with ERCP and ENBD/ERBD; 12 cases (11.7%) were treated with ERCP, needle knife fenestration and ENBD; 67 cases (65. 1% ) were treated with ERCP,EST, calculus removal and ENBD; 8 cases (7. 8% ) were treated with ERCP, EST and ERBD. Of 100 cases one (1.0%) suffered from postoperative hemorrhage of duodenal papilla, which was cured by endoscopic hemostasis. Compared with selective cases those undergoing emergency ERCP often needed two stage procedures (P = 0. 010), but there are no statistical differences between two groups in success rate (P = 0. 122). Conclusions Emergency ERCP are safe and effective in acute biliary-pancreatic diseases.