This experiment aims at exploring the prepared method of self—made fibrin glue (SMFG), its loeal hemostatic ways, curative effects and tissue reactions in the model of anticoagulated rabbits, to offer a experimental basis for clinical application. SMFG was made by a combination of swine thrombin and pyknotic fibrinogen extracted from fresh frozen plasma through double freezing, unfreezing, centrifugation—precipitation. Using SMFG in various liver wounds, local wounds were observed by gross, optical microscopic and electron microscopic ways. The results showed that SMFG had good hemostatic capacity in the model of heparinized liver trauma, without serious tissue reactions and cellular toxicity. It could satisfy the need of hemostasis in liver trauma, at the same time, it possesses some superiority of simple, safety and cheapness, and could be the best ideal alterative of commercial fibrin glue.

Objective To study the role of down regulation of bcl 2 expression of human cholangiocarcinoma cells with bcl 2 mRNA cleaving ribozyme (RZ). Methods Synthetic gene of bcl 2 mRNA cleaving RZ was recombined with the expression vector of eukaryotic cells. The recombined vector was used to transfect human cholangiocarcinoma cells. The expression of bcl 2 in human cholangiocarcinoma cells was observed by immunocytochemistry and flow cytometry. Results The expression of bcl 2 in human cholangiocarcinoma cells transfected with RZ gene was lower than that of control group. Spontaneous apoptosis was observed in a few of the cells. Conclusion bcl 2 mRNA cleaving RZ can down regulate the expression of bcl 2 of cholangiocarcinoma cells notably and can also induce spontaneous apoptosis.

The effects of targeted silencing of heparanase gene by small interfering RNA (siRNA) on invasiveness and metastasis of osteosarcoma cells (MG63 cells) were investigated in the present study. Two complementary oligonucleotide strands were synthesized and inserted into pGenesil-1 vector based on the mRNA sequence of heparanase gene. The expression vector containing short hairpin RNA (pGenesil-shRNA) was constructed successfully. MG63 cells were randomly allocated into 3 groups: blank group, empty vector (pGenesil) transfected group and expression vector (pGenesil-shRNA) transfected group. Under the induction of Lipofectamine 2000, the recombinants were transfected into MG63 cells. Heparanase gene expression level was detected by RT-PCR and Western blotting. Cell proliferation was measured by MTT assay. Cell invasiveness and metastasis were examined by cell adhesion and Transwell-ECM assays. HUVECs migration assay was applied for the detection of angiogenesis. As compared with negative controls, the mRNA and protein expression levels of heparanase were down-regulated by 76.1% (P<0.01) and 75.3% (P<0.01) respectively in the pGenesil-shRNA transfected group. Meanwhile, the proliferation, adhesiveness, invasiveness and angiogenesis properties of MG63 cells were all significantly inhibited. It was suggested that targeted silencing of heparanase gene by siRNA could dramatically inhibit the invasiveness and metastasis of osteosarcoma cells.

As a method of choice in the treatment of ampulary carcinoma,pancreaticoduodenectomy often demands open procedure.With the development of minimally invasive techniques,laparoscopic pancreaticoduodenectomy was realized in recent years.Because laparoscopic pancreaticoduodenectomy is high technique-demanding,its popularization is restricted.With flexible robotic arnls and three-dimensional imaging,Da Vinci surgical system has overcome the shortcomings of traditional laparoscope to some extent.In March 2010.a 60-year-old female patient with ampulary carcinoma underwent Da Vinci surgical system-assisted pancreaticoduodenectomy at the Southwest Hospital.The mean operation time and blood loss were 490 minutes and 450 ml respectively,and no blood transfusion was required.Out-of-bed activity began shortly after the operation.The gastric tube was removed and fluid diet was given on postoperative day 3.The patient was discharged on postoperative day 10 without incidence of complications.The Success of this case preliminarily demonstrats that Da Vinci snrgical system-assisted pancreaticoduodenectomy is safe and feasible.

Objective To investigate the clinical efficacy of laparoscopic anatomical hepatectomy via Glissonean pedicle transection approach.Methods The clinical data of 68 patients who underwent laparoscopic anatomical hepatectomy by Glissonean pedicle transection approach at the Southwest Hospital of Third Military Medical University between March 2009 and December 2013 were retrospectively analyzed.All the patients received intravenous-inhalation general anesthesia.The splitting of liver was carried out after Glissonean pedicles were completely dissected and occluded under the laparoscope according to anatomical structure.The patients with hepatolithiasis underwent intraoperative bile duct exploration and stone removal with T-tube placement based on the conditions of extrahepatic biliary lesions.The fragmented specimens from benign lesions of liver were taken out through a Trocar hole with the diameter of 12mm.The complete specimens from malignancy tumors of liver were taken out through an enlarged Trocar hole or transverse incision at the symphysis pubis.According to the recheck results of benign and malignancy lesions,the patients were followed up by outpatient examination and telephone interview till September 2014.The measurement data with normal distribution were presented as (x) ± s.The survival curve was drawn by Kaplan-Meier method.Results Among the 68 patients,laparoscopic anatomical hepatectomy via Glissonean pedicle transection approach was performed on 64 patients and 4 patients were coverted to open surgery.Laparoscopic anatomical left hemihepatectomy was performed on 30 patients,anatomical right hemihepatectomy on 19 patients,anatomical right posterior lobectomy on 10 patients and anatomical right anterior lobectomy on 9 patients.The mean operation time,volume of intraoperative blood loss,rate of perioperative blood transfusion,time of postoperative gastrointestinal function recovery and duration of hospital stay were (224 ± 117)minutes,(413 ± 349) mL,5.9％ (4/68),(3.0 ± 0.5) days and (8.0 ± 3.0) days,respectively.There was no perioperative death,and 6 patients with postoperative complication were cured by symptomatic treatment.The tumor diameter and distance to resection margin in 29 patients with malignancy tumors of liver were (4.4 ± 1.6) cm and (2.0 ± 0.9)cm,respectively.The results of pathological examination showed that hepatolithiasis was detected in 22 patients,cavernous hemangioma of liver in 12 patients,hepatic focal nodular hyperplasia in 1 patient,hepatic adenoma in 1 patient,hepatic angiomyolipoma in 1 patient,hepatic multiple cysts in 1 patient,hepatic tuberculosis in 1 patient,hepatocellular carcinoma in 27 patients and cholangiocarcinoma in 2 patients.All the 68 patients were followed up for 6-60 months with a mean time of 24 months.No recurrence and residual stones or lesions in 39 patients with benign lesions of liver were detected during the follow-up.The 1-,3-,5-year overall survival rates and tumor-free survival rates in 29 patients with malignant tumors of liver were 92％,84％,60％ and 83％,59％,42％,respectively.Conclnsion Laparoscopic anatomical hepatectomy via Glissonean pedicle transection approach is safe and feasible,and is suitable not only for left hemihepatectomy,but also for right hemihepatectomy of high technical specification.

ObjectiveTo investigate the efficacy of laparoscopic hepatectomy for regional hepatolithiasis.MethodsThe clinical data of 81 patients with regional hepatolithiasis who received laparoscopic hepatectomy at the Southwest Hospital from March 2007 to March 2011 were retrospectively analyzed.Based on the classification of the Guideline for the diagnosis and treatment of hepatolithiasis 2007 version and indications for open surgery,hepatic lobes with calculi,biliary stricture or dilated bile ducts were resected laparoscopically after preoperative examination.Bile ducts of the remnant hepatic lobes were explored using fiber choledochoscope or electronic choledochoscope for the prevention of residual stones. Results Laparoscopic hepatectomy was successfully performed on 72 patients,and the other 9 patients were converted to open surgery. Left lateral lobectomy ( segments Ⅱ,Ⅲ ) was performed on 20 patients,left hemihepatectomy ( segments Ⅱ,Ⅲ,Ⅳ ) on 30 patients,right posterior lobectomy ( segments Ⅵ,Ⅶ ) on 11 patients,right anterior lobectomy ( segments Ⅴ,Ⅷ ) on 6patients,right hemihepateetomy (segments Ⅴ,Ⅵ,Ⅶ,Ⅷ ) on 9 patients,hepatic Ⅲ segmentectomy on 2 patients and hepatic Ⅵ segmentectomy on 3 patients.Gallbladders were resected,and intermittent portal triad clamping was performed on 15 patients.Hepateetomy combined with biliary exploration and stone removal was performed on 57 patients.The mean operation time and operative blood loss were (328 ± 80)minutes and (451 ±288) ml,respectively.No operative mortality was observed.Sixteen patients had postoperative complications,including pulmonary infection in 2 patients,pleural effusion in 4 patients,encapsulated effusion in the resection margin in 6 patients,abdominal infection and abscess in 2 patients and wound infection in 2 patients.Of the 16patients with complications,9 were cured by medicine,6 by pleural or abdominal drainage and 1 by wound debridement.The mean duration of hospital stay was ( 13 + 6)days.The intraoperative stone clearance rate was 96％ (69/72),and the residual stone in 3 patients were removed by choledochoscopy.Sixty-nine patients were followed up for 7-55 months,7 patients had symptoms of cholangitis and 2 patients had common bile duct stone recurrence.ConclusionsLaparoscopie hepatectomy is sate and effective for regional hepatolithiasis.Accurate positioning of the stones and lesions pre- and intra-operatively,reasonable designing of the parenchymal transection plane,and anatomical liver resection are the key points for achiving good therapeutic effects.

Objective To investigate the indications, techniques and results of laparoscopic hepatectomy. Methods The clinical data and follow-up results of 463 patients who received laparoscopic hepatectomy at our institute were retrospectively analyzed. Results From March 1, 2007 to March 31, 2011, 463 cases of laparoscopic hepatectomy were successfully carried out. Of the 463 patients,165 were with primary liver cancer, 29 with metastatic liver cancer, 143 with hepatic hemangioma, 81with hepatolithiasis and 45 with other benign liver diseases (including hepatic angiomyolipoma, hepatocellular adenoma, focal nodular hyperplasia and chronic liver abscess). The surgical approaches included laparoscopic left lateral lobectomy (93 cases), left hepatectomy (71 cases), extended left hepatectomy (4 cases), right hepatectomy (29 cases), right posterior lobectomy (24 cases), hepatectomy of segment Ⅵ (56 cases), extended right hepatectomy (2 cases), central hepatectomy (8 cases) and hepatectomy of segments Ⅶ/Ⅷ, Ⅳa, caudate lobe and the junction of segment Ⅵ and Ⅶ (41 case).Nonanntomic and wedge resection were performed on 121 patients, and combined resection on 14 patients. The mean operation time, blood loss, length of hospital stay and incidence of postoperative complications were (244.71 ± 105. 07) minutes, (460. 26±425.81) ml, (15.51 ±4.36) days and 9.29%, respectively. And no operative death occurred. In the 194 cases with malignant liver lesions,185 cases were followed up for 2 to 50 months. The 1 year and 3 year overall and disease free survival rate were 90. 8% and 87.9% , 84.2% and 73. 7% respectively. Conclusions As a means of minimally invasive surgical approach, laparoscopic hepatectomy can be selectively adopted for the treatment of all kinds of liver diseases which located at different parts of the liver, with the advantages of smaller trauma, quick recovery and cosmetic benefits. The short-term results of laparoscopic hepatectomy is superior to and its long-term results is equal to that of open surgery. Benign liver diseases, small hepatocellular carcinoma and metastatic liver cancer are the good indications for laparoscopic hepatectomy.

Objective To investigate the value of laparoscopy in the treatment of liver cancer.Methods The clinical data of 128 liver cancer patients who received laparoscopic surgery at Southwest Hospital from March 2007 to October 2009 were retrospectively analyzed.Of all patients,116 were with primary liver cancer,and 12 with metastatic liver cancer.There were 107 patients who received laparoscopie bepatectomy,15 received laparoscopic radiofrequency ablation(RFA)and 6 received laparoscopic ligation of the right branch of portal vein.Results Of the 107 patients who received laparoscopic bepatectomy,7 were converted to open surgery,and 5 were converted to hand-assisted laparoscopic hepatectomy.Anatomical hepatectomy was performed on 88 patients,including left lateral lobectomy on 21,left hemihepatectomy on 15,extended left hemihepatectomy on 2,medial lobectomy on 1,right hemihepatectomy on 11,right posterior lobeetomy on 9 and hepatic segmentectomy on 29.Combined hepatic resection was performed on 4 patients,and nonanatomical hepatectomy on 15.The mean oporatire time and blood loss were(228±92)minutes and(393±213)ml,with no operative mortality.The mean postoperative hospital stay was(8±4)days,and the incidence of complications was 15%(16/107).A total of 126 patients were followed up for 1-42 months,12 patients with laparoscopic hepatectomy died within 16 months,with the mean survival time of(118±7)weeks and the mean tumor free survival time of(105±7)weeks;2 patients with laparoscopic RFA died within 11 months:2 patients with laparoseopie ligation of the right branch of portal vein received two-stage radical resection.Conclusion Laparoscopic surgery is safe and feasible with the advantages of minimal operative trauma and quick recovery of patients when it is applied to the treatment of liver cancer.

Objective To evaluate the safety and feasibility of laparoscopic left hepatectomy in patients with primary hepatic carcinoma. Methods Retrospective analysis was conducted in clinical data of left hepatectomy from 2007 to 2014,including 47 cases of laparoscopic hepatectomy and 46 cases of open hepatectomy. The intraoperative and postoperative data of the two surgical methods were compared to evalu-ate whether laparoscopic left hepatectomy is safe and feasible. Results The tumor size were bigger in the open hepatectomy [(56. 57 ± 24. 56) mm vs. (64. 11 ± 33. 39) mm,P=0. 218]. The laparoscopic left hepatectomy resulted in shorter operation time [(217. 53 ± 60. 22) min vs.(306.80±119.91)min],andtherewasasignificantlydifference(P<0.05).Theintraoperativebloodlosswerelesserinthelaparo-scopic left hepatectomy [(350. 21 ± 197. 98) mL vs. (556. 74 ± 471. 41) mL],and there was a significantly difference (P<0. 05). The lap-aroscopic left hepatectomy had a smaller intraoperative blood transfusion rate (12. 8% vs. 32. 6%,P<0. 05). The length of ICU stay,time for gastroentestinal function recovery, postopetative hospital stay were shorter than those of open left hepatectomy (P<0. 05). There was no significant difference of postoperative complication rate between them (14. 9% vs. 23. 9%,P=0. 271). The survival rates of 1-and 3-year after operation in patients with laparoscopic left hepatectomy were 91. 5% and 83. 0% respectively,while 84. 8% and 76. 1% in patients un-derwent open hepatectomy. The tumor-free survival rates 1-and 3-year after operation were 74. 5% and 59. 6% in patients with laparoscopic left hepatectomy respectively,while 65. 2% and 54. 3% in patients underwent open hepatectomy. Conclusion Laparoscopic left hepatectomy for primary hepatic carcinoma is safe and feasible. Laparoscopic left hepatectomy could be a consideration as the standard surgical methods for hepatic carcinoma.