Abstract:Abstract:Objective:To explore the effective therapeutic methods for hepatolithiasis.
Methods :We retrospectively analyzed the clinical material of 178 hepatolithiasis patients who underwent surgical therapy. Among the 178 patients, 107 patients were treated with T- tube drainage(including 7 patients with left hepatic lobe or quadlrate lobe resection), and 71 patients were treated with bile duct-intestinal anastomosis (including 11 patients with left hepatic lobe or quadrate lobe resection).
Results:Recurrence rate of postoperative angiocholitis in the two groups was 49.5 % and 24 % respectively, and the reoperation rate was 28.7 % and 8 % respectively.
Conclusions:The key to prevent recurrent angiocholitis and reduce the reoperation rate is to relieve biliary tract stricture, remove the focus of infection and provide unobstructed bile duct drainage.

Abstract:Abstract:Objective:To investigate the value of clinical use of laparoscopy combined with choledochoscopy in treatment of cholecystolithiasis complicated with choledocholithiasis.
Methods :Of 120 patients suffering from cholecystolithiasis combined with choledocholithiasis,56 patients underwent laparoscopic cholecystectomy together with choledocholithotomy and T-tube drainage(LCTD), and 64 patients received routine cholecystectomy and choledocholithotomy plus T-tube drainage by laparotomy. Postoperative administration of analgesics, wound infection, recovery of function of gastrointestinal tract (RFGI), and hospital stay were compared between the two groups.
Results:In both groups, there were no severe complications. Wound infection rate was 12.5 %, average hospital stay was （14.9±3.1） d, and RFGI was （26.1±8.6） h in routine operation group; while in LCTD group, there was no wound infection occurred, hospital stay was （5.8±1.4） d and RFGI was （10.6±3.4） h. There were significant difference in postoperative recovery conditions between the two groups (P< 0. 05~0.01). After 1 month to 3 years follow - up, 2 patients suffered from recurrence of choledocholithiasis in routine operation group.
Conclusions:Laparoscopic cholecystectomy together with choledocholithotomy and T-tube drainage, has the advantages of minimal invasion,quick recovery and short hospital stay with safe and effective outcomes, and, depending on the circumstances, can be selected as a therapeutic choice for cholecystolithiasis complicated with choledocholithiasis.

Abstract:Abstract:Objective:To compare the clinical results of choledocholithotomy by laparoscopic surgery and open surgery.
Methods: One hundred and Sixty-nine patients with cholecystoithiasis combined with choledocholithiasis were divided into laparoscopic surgery group(85 cases of laparoscopic choledocholithotomy, LCD-TD) and open surgery group (84 cases of open choledocholithotomy, OCH-TD). The clinical data in the two groups were compared and analyzed.
Results:The amount of intraoperative bleeding, the recover time of intestinal peristalsis after treatment, length of hospital stay, cases of postoperative pain and biliary fistulas in laparoscopic surgery group were evidently better than those of open surgery group(all P<0.05). No residual calculi were found on T-tube radiogram in both groups.
Conclusions:LCH-TD surgery is minimally traumatic, has good clinical results, and in most cases, can replace open choledocholithotomy.

Abstract:Abstract:Objective:To evaluate the practical value of ERCP for etiologic diagnosis and treatment of postcholecystectomy syndrome(PCS).
Methods :A retrospective review of 116 cases of postcholecystectomy syndrome received ERCP in the recent four years in our hospital was undertaken. Of the116 cases, 80 cases (68.97 %) had the indications for therapeutic ERCP and were treated by endoscopy.
Results:The bile duct was visualized in all of the 116 cases (100 %), and the pancreatic duct was visualized in 97 cases (83.62 ％). The etiology of PCS was choledocholithiasis in 56 cases (48.28 ％), diverticulum adjacent to duodenal papilla or papilla with in the diverticulm in 19 cases (16.37 ％),residual of long cystic duct with stones in 15 cases (12.93 ％), benign stenosis of terminal commom bile duct in 13 cases (11.20 ％), sphincter of oddi dysfuction in 4 cases (3.45 ％), tumour of doudenal papilla in 3 cases (2.59 ％), stenosis of injured bile duct in 2 cases(1.73 ％), sclerosing cholangitis in one case (0.86 ％), cholangiocarcinoma in one case (0.86 ％), remnant gallbladder with stones in one cases (0.86 ％), and chronic pancreatic ductal lithiasis in one case (0.86 ％). Fifty-seven cases were treated by EST stone extraction, 20 cases by ERBD,23 cases by ENBD, 1 case by EMBE, and 1 case by EPS extraction.In the 80 cases treated with endoscopy,75 cases were followed up for 3~12 months, with relief of symptons in 74 cases(98.67 ％).
Conclusions:ERCP is the gold standard for etiologic diagnosis of PCS. PCS is an indication for therapy by ERCP. Therapeutic ERCP is the preferred treatment of PCS cases with an indication for endoscopic treatment.

Abstract:Abstract:Objective:To investigate the diagnosis and surgical treatment of Mirizzi syndrome.
Methods :The clinical data of 35 cases of Mirizzi syndrome treated by surgery were analyzed retrospectively.
Results:The 35 patients included 13 males and 22 females, with mean age of 62.3 years. Forteen cases (40.0 %) were diagnosed as Mirizzi syndrome preoperatively, and 16 cases (60.0 %) were recognized during operation. According to Csendes classification, 9 cases were type Ⅰ, 18 cases type Ⅱ，7 cases type III and 1 case type Ⅳ. Laparoscopic operation was performed in 13 patients, among them, 4 cases（30 %）converted to open laparotomy. The laparoscopic operations included cholecystectomy in 7 cases, and partial cholecystectomy plus intraoperative cholangiography in 2 cases. Tweenty-six patients underwent open laparotomy, including cholecystectomy in 9 cases, partial cholecystectomy plus T-tube drainage in 17 cases, and Roux-en-Y hepaticojejunostomy in 2 cases. Postoperative complications included biliary leakage in 3 cases, infection of incisional wound in 1 case and pneumonia in 1 case, all the complications recovered by non-surgical treatment. Thirty-three cases were followed-up for 1-5 years, the results were satisfactory.
Conclusions:The preoperative diagnosis and intraoperative identification of Mirizzi syndrome, especially during difficult laparoscopic cholecystectomy, should be emphasized, and timely conversion to open surgery could decrease the risk of bile duct injury. Partial cholecystectomy plus intraoperative cholangiography can be helpful to identify pathological change and to reduce the risk of iatrogenic biliary duct injury of Mirizzi syndrom. In the event of serious destruction of biliary duct, Roux-en-Y hepaticojejunostomy is feasible.

Abstract:Abstract:Objective:To investigate the indications and appropriate method of early operative treatment of gallstone pancreatitis(GP).
Methods :The clinical data of 136 cases of GP that underwent early operation were retrospectively analyzed.
Results:(1) Among 10 cases with severe obstructive GP, 8 cases underwent early operation, and all of them were cured. (2) Among 70 cases with mild obstructive GP, 57 cases underwent early operation, and all of those cases were cured. （3）In 47 cases with severe non-obstructive GP, 13 cases underwent simple early operation, and 4 of those cases died of early multiple organ failure, while 3 other cases treated by regional intra-arterial infusion combined with early operation were cured. （4）In 146 cases with mild non-obstructive GP, 55 cases underwent early operation, and all of those cases were cured.
Conclusions:（1）Obstructive GP is an absolute indication for early operation, and the basic operation is decompression of biliary tract obstruction and peri-pancreatic drainage. （2）In principle, cases of severe non-obstructive GP should not undergo early operation. （3）Early operation might be carried out in cases of mild non-obstructive GP.

Abstract:Abstract:Objective:To study the action of E2F1 protein signal transmission pathway to influence the apoptosis of cholangiocarcinoma QBC939 cells.
Methods :Four siRNAs were designed according to the coding sequence of E2F1 gene, and cloned into the downstream of U6 promoter of pGenesil. The constructed recombinant was analyzed and identified by AseⅠ endonuclease digestion and DNA sequencing. The siRNA constructs were transfected into QBC939 cells via lipofectamine 2000. RT-PCR was used to measure the E2F1 mRNA expression in QBC939, and the rate of apoptosis of QBC939 was detected by the method of flow cytometry after transfection.
Results:The constructed psiRNA plasmid digested with AseⅠwas linearized. The sequencing result confirmed that the sequence of inserted fragment was correct. The expression of E2F1 mRNA was greatly inhibited at 48h after transfection, and the rate of apoptosis of QBC939 increased significantly after transfection.
Conclusions:Eukaryotic expression vector of siRNA targeting E2F1 gene can specifically inhibit E2F1 expression and promote the apoptosis of QBC939 cell. It can be used for later experimental research in cholangiocarcinoma.

Abstract:Abstract:Objective:To investigate the expression of stem cell factor Oct-4 in human cholangiocarcinoma QBC939 subclone cell lines with different metastatic capacities.
Methods :The screening and use of one subclone cell line with high metastasis capacity and one subclone cell line with low metastasis capacity was performed by the in vitro invasive assay. Then, the biologic characters of the two subclone cell lines were compared, analyzed and identified. The expression of the stem cell factor Oct-4 in the two selected subclone cell lines was determined by RT-PCR and Western-blot assays.
Results:Two subclones of QBC939 with different metastatic capacities, and named QBC-H and QBC-L were isolated，and their invasion indexes were 263±5.9 and 154±6.6, respectively (P<0.05）. The expression of mRNA and protein of Oct-4 was stronger in QBC-H than in QBC-L.
Conclusions:There is a close relationship between the expression strength of the stem cell factor Oct-4 and the metastatic capacity of Oct-4.The stem cell factor may play an important role in regulation of the metastatic capacity of cholangiocarcinoma.

Abstract:Abstract:Objective:To elucidate the mechanisms of disruption of intestinal mucosa barrier in obstructive jaundice.
Methods :The obstructive jaundice model of rats was set up.At 10 d and 20 d after operation,immunohistochemistry and Western-blot techniques were used to examine the distribution and expression of tight junction proteins (ZO-1,Occludin) and myosin light chain kinase（MLCK）in intestinal mucosa.
Results:In normal control groups, the staining of ZO-1 and occludin was predominantly localized to the margins of the epithelial cells and the apex of the cell membrane, and displayed a continuous and uniform distribution along the under surface of the villae. MLCK was distributed mainly in cytoplasm.In obstructive jaundice groups, ZO-1 and occludin staining appeared discontinuous and vague,with rough edges and spiculate processes.The staining of MLCK was also discontinuous and scanty.The strong positive express ratio of ZO-1,Occludin and MLCK were obviously lower in two experiment group than those in the control group all P＜0.05). ZO-1 was significantly decreased in 20 d group than that in 10 d group（14.3 % vs28.6 %，P＜0.05), but no significant difference in occludin and MLCK staining between 10 d group and 20 d group.Similar outcomes were obtained by quantitative analysis of Western blot images.
Conclusions:Obstructive jaundice rats have distribution of ZO-1,occludin and MLCK in ileum mucous membrane epithelium, and is associated with breakdown of the integrity of the intestinal barrier.

Abstract:Abstract:Objective:To study the dynamics of plasma endotoxin(PE) levels in the perioperative period of patients with malignant obstructive jaundice, and the significance of hydrogen breath test (HBT) in judging of small intestinal bacterial translocation.
Methods :Thirty-eight patients with malignant obstructive jaundice who were suitable for operation, were defined as treatment group (group A, n=38) and 30 healthy individuals were defined as control group (group B, n=30). The changes of PE levels and the results of HBT during the perioperartive period were observed in group A, and compared with group B.
Results:On admission to hospital, the PE level in group A ［(0.69±0.22) EU/mL］ and group B ［（0.17±0.05) EU/mL］ was significant difference （P＜0.01）. During the 1 d preoperative period, the PE levels decreased a little（P＞0.05）, but it decreased significantly after operation（all P＜0.05）. The positive rates of HBT in group A at admission, 1 d preoperative and 3 d, 7 d and 14 d after operation, was 86.8％, 73.7％, 39.5 %, 21.1 % and 7.9 %; and it was 6.7 % in group B. The difference was significant between group A before operation and group B; and in group A, comparing postoperative 3 d and 7 d values with preoperative 1 d value, the differences were significant (P＜0.05).
Conclusions:The patients with malignant jaundice have small intestinal bacterial translocation and have endotoxemia to some extent. HBT is a valid index for judging bacterial translocation in the perioperative period of patients with malignant obstructive jaundice.

Abstract:Abstract:Objective:To investigate the interfering effect of the small hairpin RNA (shRNA) targeted to human telomere retrotranscriptase (hTERT) gene and its silence effect on telomerase hTERT gene expression in HeLa cells.
Methods :After preparation of the shRNA in-vitro, the shRNA was transfected to HeLa cells by the calcium phosphate method. Telomerase activity was detected by TRAP-silver stain and PCR-EIA.
Results:The telomerase activity of HeLa cells decreased significiantly after transfected with the 46-base small hairpin RNA.
Conclusions:shRNA (targeted to human telomerase hTERT gene) prepared by this method displays a significant silence effect on telomerase gene expression. It could be a new approach of obtaining shRNA for RNAi research in tumor inhibition.

Abstract:Abstract:Objective:To investigate the effects of human arresten gene transfection on the intimal hyperplasia of venous autografts in vivo.
Methods :Autogenous vein graft model was established in male Sprague-Dawley rats. The transplanted veins were immersed in the liposome mediated recombinant eukaryotic expression plasmid pSecTag2-AT solution(group Ⅰ) or the liposome mediated control plasmid pSecTag2 solution(group Ⅱ) for 30minutes just before vascular anastomosis. Normal control group(group Ⅲ) was only treated with isovolume of liposome. Vein graft samples were harvested at 4 weeks after operation. RT-PCR was used to detect the expression of arresten mRNA in blood vessel; the surface area and thickness of the intima and media were measured by computerized planimetry under a light microscope to compare the degree of neointimal hyperplasia by the calculated ratio between intima (I) and media (M) after staining with hematoxylin-eosin and Verhoeff(elastic fibers). Immunohistochemical labeling and morphologic analysis of vein graft sections were used to identify PCNA positive cells and α-SMA positive cells; Western blot was used to detect the protein of TGF-β1.
Results:The genome of arresten-transferred tissue contained a 449bp specific fragment of arresten gene was expressed in group Ⅰ， but was not expressed in group Ⅱ or group Ⅲ; the surface area of the intima and media of group Ⅰ was less than that of group Ⅱ and group Ⅲ, and the difference were statistically significant (P<0.05), while I/M had no statistical difference (P>0.05). A less intimal thickness of groupⅠ was seen compared with group Ⅱ and group Ⅲ (P<0.01). α-SMA staining suggested that VSMC was present in the hyperplastic intimal. The number of PCNA-positive-stained cells and expression index of group Ⅰ was lower as compared with that of group Ⅱ and group Ⅲ (P<0.05). Protein level of TGF-β1 of group Ⅰ decreased obviously compared with group Ⅱ and group Ⅲ.
Conclusions:Transfection of human arresten gene can effectively inhibit the intimal hyperplasia of venous autograft. Arresten has a potential perspective for clinic application in prevention and treatment of restenosis after vascular transplantation.

Abstract:Abstract:Objective:To investigate the cytotoxicity of ADM-PBCA-NP on L-02 cells.
Methods : L-02 cells were cultured in vitro and the LDH activity of supernatant liquid of culture cells was examined. The toxicity of ADM-PBCA-NP, ADM and PBCA-NP to L-02 cells by the MTT assay was also determined, and the haemolysis function of PBCA-NP with different concentrations was detected.
Results:The cytotoxicity of ADM-PBCA-NP, ADM and blank PBCA nanoparticles to L-02 cells under the 10-6 mol/L concentration range was not cytototic (grade 1). LDH activity of supernatant liquid of culture cells showed no differences between the 3 groups.
Conclusions:Nanoparticles of ADM-NP and PBCA-NP in the 10-6 mol/L concentration range have no significant toxic effect on L-02 hepatic cells; and in a certain concentration range, the cell compatibility is excellent and will not lead to hemolysis.

Abstract:Abstract:Objective:To explore a method to improve the radical resection and survival rate of hilar cholangiocarcinoma.
Methods :Fifty-eight consecutive patients with hilar cholangiocareinoma who underwent anatomic hepatectomy together with caudate lobectomy from Feb 1998 to June 2006 were studied retrospectively.
Results:Tumor resection combined with anatomic left hemihepatectomy was performed in 26 patients, combined with anatomic right hemihepatectomy in 25 patients, with right hepatic trisectionectomy in 4 patients, and with middle hepatectomy in 3 patients. The entire caudate lobe was resected and skeletization lymph node dissection of the hepatoduodenal ligament was performed in all cases. The radical resection rate was 72.4 ％(42/58), and the 1-,and 3-year survival rate was 79.5 ％, and 36.2 ％ respectively. At present, 3 cases have tumor-free survival for 6-year.
Conclusions:Tumor resetion combined with anatomic hepatectomy and caudate lobectomy can offer a better chance of long-term survival in some selected patients with advanced hilar cholangiocarcinoma. Skilled surgical technique can result in better outcomes.

Abstract:Abstract:Objective:To explore the technique of T-tube replacement after choledochoscopy.
Methods :Choledochoscopy was performed in 256 cases, replacement of T-tube was conducted by direct insertion technique in 85 cases (Group A),and by a wire-guide technique in 171 cases(Group B).
Results:Three cases of T-tube replacement were unsuccessful in group A, in one case of which, the T-tube was misguided into the bowel and the patient recovered after conservative treatment following the withdrawal of T-tube. In group B, all the T-tube replacements were conducted successfully with the wire-guide technique.
Conclusions:Replacement of T-tube is facilitated by the wire-guide technique.

Abstract:Abstract:Objective:To review the clinical features and early management of combined thorcoabdominal injury (CTI), to decrease the mortality rate.
Methods :A retrospective study was carried out in 63 cases of CTI treated surgically from May 1990 to May 2007.There were 34 cases with blunt injuries and 29 cases with penetrating injuries.Forty-seven patients had rapture of left diaphragm, 9 cases had rupthure of right diaphragm, 7 cases had rupture of both diaphragms, 24 cases were combined with traumatic diaphragmatic hernia, 63 cases were combined with hemopneumothorax, and 60 cases were combined with shock. Forty-one cases were diagnosed correctly before surgery. Sixty-three cases were treated by surgery, among them, surgery was performed through left thoracotomy in 18 cases, through a right thoracotomy in 2 cases, through bilateral thoracotomy in 5 cases, through thoraco-abdominal incision in 2 cases, and through the abdominal route in 32 cases.
Results:Fifty-six cases were cured, and 7 cases (11.1 %) died. The main cause of death was hemorrhagic shock.
Conclusions:CTI has high mortality rate, and the most frequent cause of mortality is hemorrhagic shock.The operative approach is based on the individual injury condition.Early diagnosis and timely surgical intervention is the key to improving the cure rate and reducing mortality in severe CTI.

Abstract: