[Experimental study on intervention effect of Grifola frondosa on nonalcoholic steatohepatitis].

To study the preventive effect of Grifola frondosa on nonalcoholic steatohepatitis (NASH). The rat model of NASH was established by feeding high-fat diets for 12 weeks and intervened with 0.5 g · kg(-1) · d(-1) and 1.0 g · kg(-1) · d(-1) of C. frondosa powder suspensions. The degrees of hepatocyte fatty degeneration and inflammation were observed under the optical microscope with routine HE staining. The NAFLD activity scores (NAS) were calculated. Serum ALT, AST and hepatic TG and CHOL were tested by the biochemical method. The hepatic MDA was examined by thiobarbituric acid method. The hepatic SOD was tested by the xanthine oxidase test. The hepatic GSH-PX activity was determined by the dithio-nitrobenzoic acid method. Hepatic TNF-α and IL-6 were detected by the enzyme-linked immunosorbent assay (ELISA). The NASH model group induced by high-fat diets showed higher hepatic NAS, ser- um ALT, AST, CHOL and hepatic TG, CHOL, MDA, TNF-α, IL-6 (P < 0.01 or P < 0.05) and lower serum TG and hepatic SOD, GSH-PX (P < 0.01, P < 0.05) than the normal control group. After being intervened with different doses of G. frondosa, the NASH group revealed significantly lower hepatic NAS, serum ALT and hepatic TG, CHOL, MDA, TNF-α and IL-6 (P < 0.05) and higher hepatic SOD, GSH-PX (P < 0.05) than the model group. G. frondosa may prevent the further development of NASH by improving the disorder of lipid metabolism in rats with NASH induced by high-fat diets, relieving the level of oxidative stress and reducing the generation of inflammatory cytokines.

Use of an autologous liver round ligament flap zeros postoperative bile leak after curative resection of hilar cholangiocarcinoma.

BACKGROUND: Postoperative bile leak is a major surgical morbidity after curative resection with hepaticojejunostomy for hilar cholangiocarcinoma, especially in Bismuth-Corlette types III and IV. This retrospective study assessed the effectiveness and safety of an autologous hepatic round ligament flap (AHRLF) for reducing bile leak after hilar hepaticojejunostomy. METHODS: Nine type III and IV hilar cholangiocarcinoma patients were consecutively hospitalized for elective perihilar partial hepatectomy with hilar hepaticojejunostomy using an AHRLF between October 2009 and September 2013. The AHRLF was harvested to reinforce the perihilar hepaticojejunostomy. Main outcome measures included operative time, blood loss, postoperative recovery times, morbidity, bile leak, R0 resection rate, and overall survival. RESULTS: All patients underwent uneventful R0 resection with hilar hepaticojejunostomy. No patient experienced postoperative bile leak. CONCLUSIONS: The AHRLF was associated with lack of bile leak after curative perihilar hepatectomy with hepaticojejunostomy for hilar cholangiocarcinoma, without compromising oncologic safety, and is recommended in selected patients.

Nanoparticle-mediated local delivery of an antisense TGF-ß1 construct inhibits intimal hyperplasia in autogenous vein grafts in rats.

BACKGROUND: Intimal hyperplasia is one of the most important causes of vascular graft failure. Numerous studies have correlated transforming growth factor-ß1 (TGF-ß1) with extracellular matrix (ECM) deposition, a hallmark of intimal thickening. PRINCIPAL FINDINGS: In the present study, we performed immunohistochemistry, RT-PCR, and Western blot to examine the dynamic expression of TGF-ß1, TGF-ß1 receptor type I (TGF-ß RI), matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) during intimal hyperplasia in grafted veins of a rat model generated by grafting a portion of the right internal jugular vein to the ipisiliary carotid artery. Additionally, we determined whether nanoparticle-mediated delivery of a TGF-ß1 antisense-expressing construct prevented TGF-ß1 expression and intimal hyperplasia in grafted veins. In grafted veins, the expression of TGF-ß1 significantly increased on day 3 after transplantation, peaked on day 7, slightly decreased on day 14, and returned to baseline levels on day 28. The positive expression of TGF-ß RI in grafted veins remarkably increased on day 7, peaked on day 14, and decreased thereafter. MMP-1 expression decreased significantly, while TIMP-1 expression increased, significantly on days 14 and 28. Nanoparticle-mediated delivery of a TGF-ß1 antisense-expressing construct down-regulated TGF-ß1 expression and inhibited intimal hyperplasia in grafted veins. CONCLUSIONS: Our findings provide further evidence that TGF-ß1 plays an integral role in the development of intimal hyperplasia after vascular injury. Nanoparticle-mediated delivery of a TGF-ß1 antisense-expressing construct is a feasible strategy to target TGF-ß1-induced intimal thickening.

Abdominal separation in an adult male patient with acute abdominal pain.

We report a male patient with prolonged post-prandial abdominal distension and a sudden onset of epigastric pain initially diagnosed as acute abdomen. The patient had no history of surgery. Physical examination revealed peritonitis and abdominal computed tomography scan showed upper abdominal mesentery intorsion. The patient then underwent surgical intervention. It was found that the descending mesocolon dorsal root was connected to the ascending colon and formed a membrane encapsulating the small intestine. The membrane also formed an orifice in the ileal pars caeca, from which a 30 cm herniated ileum formed a "C"-shaped loop which was strangulated by the orifice. An abdominal separation was diagnosed after surgery. We liberated the membranous peritoneum which incarcerated the intestinal canal from the root of ileocecal junction to Treitz ligament, and reduced the small intestinal malrotation. The patient had an uneventful recovery after operation with his abdominal distention disappeared during the follow-up. Abdominal separation is a rare situation, which may be related with embryo development. Surgery is a choice of treatment for it.

Induction of biliary cholangiocarcinoma cell apoptosis by 103Pd cholangial radioactive stent gamma-rays.

BACKGROUND: In recent years, interventional tumor therapy, involving implantation of intra-cholangial metal stents through percutaneous trans-hepatic punctures, has provided a new method for treating cholangiocarcinoma. (103)Pd cholangial radioactive stents can concentrate high radioactive dosages into the malignant tumors and kill tumor cells effectively, in order to prevent re-stenosis of the lumen caused by a relapsed tumor. The aim of the present study was to investigate the efficacy of gamma-rays released by the (103)Pd biliary duct radioactive stent in treating cholangiocarcinoma via induction of biliary cholangiocarcinoma cell apoptosis. METHODS: A group of biliary duct cancer cells was collectively treated with a dose of gamma-rays. Cells were then examined by the 3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl terazolium-bromide (MTT) technique for determining the inhibition rate of the biliary duct cancer cells, as well as with other methods including electron microscopy, DNA agarose gel electrophoresis, and flow cytometry were applied for the evaluation of their morphological and biochemical characteristics. The growth curve and the growth inhibition rate of the cells were determined, and the changes in the ultrastructure of the cholangiocarcinoma cells and the DNA electrophoresis bands were examined under a UV-lamp. RESULTS: The gamma-ray released by (103)Pd inhibited cholangiocarcinoma cell growth, as demonstrated when the growth rate of the cells was stunned by a gamma-ray with a dosage larger than 197.321 MBq. Typical features of cholangiocarcinoma cell apoptosis were observed in the 197.321 MBq dosage group, while cell necrosis was observed when irradiated by a dosage above 245.865 MBq. DNA agarose gel electrophoresis results were different between the 197.321 MBq irradiation dosage group, the 245.865 MBq irradiation dosage group, and the control group. CONCLUSIONS: (103)Pd radioactive stents which provide a radioactive dosage of 197.321 MBq are effective in the treatment of cholangiocarcinoma; (103)Pd radioactive stents should be useful for the clinical treatment of cholangiocarcinoma.

Hypermethylation and aberrant expression of secreted frizzled-related protein genes in pancreatic cancer.

AIM: To determine the methylation status and aberrant expression of some secreted frizzled-related protein (SFRP) genes in pancreatic cancer and explore their role in pancreatic carcinogenesis. METHODS: Methylation status and expression of SFRP genes were detected by methylation-specific PCR (MSPCR) and reverse-transcription PCR (RT-PCR) respectively. RESULTS: The frequencies of methylation for SFRP genes 1, 2, 4, 5 were 70%, 48.3%, 60% and 76.7% in pancreatic cancer samples, and 21.7%, 20%, 10% and 36.7% in matched cancer adjacent normal tissue samples, respectively (c2 = 28.23, P < 0.0001 for SFRP gene 1; c2 = 10.71, P = 0.001 for SFRP gene 2; c2 = 32.97, P < 0.0001 for SFRP gene 4; c2 = 19.55, P < 0.0001 for SFRP gene 5). Expression loss of SFRP genes 1, 2, 4 and 5 was found in 65%, 40%, 55% and 71.7% of 60 pancreatic cancer samples, and 25%, 15%, 18.3% and 31.7% of matched cancer adjacent normal tissue samples, respectively (c2 = 19.39, P < 0.0001 for SFRP gene 1; c2 = 9.40, P = 0.002 for SFRP gene 2; c2 = 17.37, P < 0.0001 for SFRP gene 4; c2 = 19.22, P < 0.0001 for SFRP gene 5). SFRP gene 1 was methylated but not expressed in PC-3 and PANC-1, SFRP gene 2 was methylated but not expressed in PANC-1 and CFPAC-1, SFRP gene 4 was methylated but not expressed in PC-3, and SFRP gene 5 was methylated but not expressed in CFPAC-1. CONCLUSION: Hypermethylation and aberrant expression of SFRP genes are common in pancreatic cancer, which may be involved in pancreatic carcino-genesis.

103Pd-induced apoptosis of proliferative smooth muscle cells in bile ducts of dogs: significance and effects on related genes.

BACKGROUND: With the objective of developing a locally-produced radioactive stent, the present study used in vivo animal experiments to explore apoptosis of proliferative smooth muscle cells resulting from facilitation of the expression of genes caused by gamma-radiation in order to prevent bile duct restenosis. We therefore explored the effects and significance of gamma-radiation on the activity of caspase-3, Fas and Bcl-2 genes in apoptosis of proliferative smooth muscle cells in the bile duct walls of dogs. METHODS: Twelve dogs were randomly divided into 2 groups (6 in each group). A postinjury bile duct stenosis model was established and radioactive (103)Pd ((103)palladium) or ordinary bile duct stents were implanted into the bile ducts. HE staining, RT-PCR and immunohistochemistry were used to detect the proliferation and apoptosis of bile duct smooth muscle cells in proliferative endomembrane and the expression of related caspase-3, Bcl-2 and Fas genes. RESULTS: The expression of caspase-3 and Fas genes in the bile duct tissues of dogs with radioactive stents was higher than that of dogs with ordinary stents. There was significant apoptosis of proliferative smooth muscle cells in the bile ducts. The expression of the Bcl-2 gene in the bile duct tissues of dogs with radioactive stents was lower than that in those with ordinary stents. There was significant apoptosis of proliferative smooth muscle cells in the dogs with low Bcl-2 gene expression. CONCLUSIONS: Radiation increases the activity of caspase-3 and Fas genes and is associated with apoptosis. The radioactive (103)Pd stent may facilitate apoptosis of proliferative smooth muscle cells in the bile ducts of dogs by activating these genes. The Bcl-2 gene expression level is correlated with the occurrence of apoptosis and the radiosusceptibility of cells.

[Determination of intestinal permeability in cholelithiasis patients by oral administration of technetium-99m-diethylenetriaminepentaacetatic acid].

OBJECTIVE: To investigate the intestinal permeability of patients with cholelithiasis of different types. METHODS: Technetium-99m-diethylenetriaminepentaacetatic acid (99mTc-DTPA) at the dose of 185 MBq (5 mCi) was administered orally to 56 patients of cholelithiasis, 15 cases of cholesterol stone (CS group) and 41 cases of pigment stone (PS group) based on the cross section of the stone during operation, and 17 healthy controls. A 24 h urine collection was obtained after the ingestion of the tracer to calculate the urinary excretion of DTPA. RESULTS: The mean percentage of the total ingested dose of 99mTc-DTPA excreted in a 24 h urinary excretion was 5.0%+/-3.6% in the CS group, not significantly different from that in the control group (4.5%+/-3.4%. F=2.18, P>0.05), and the mean percentage of the total ingested dose of 99mTc-DTPA excreted in a 24 h urinary excretion of the PS group was 10.5%+/-6.9%, significantly higher than that in the control group (F=7.62, P<0.05), showing a significantly increase of intestinal permeability (P<0.05). CONCLUSION: The intestinal permeability of the patients of pigment stone is higher than that of the healthy subjects. Hyperpermeability may be a factor of the pathogenesis of pigment stone.

103Pd radioactive stent inhibits biliary duct restenosis and reduces smooth muscle actin expression during duct healing in dogs.

BACKGROUND: This study was designed to assess the expression of smooth muscle actin (SMA) in the healing process after implanting a (103)Pd radioactive stent in the biliary duct, and to discuss the function and significance of this stent in preventing biliary stricture formation. METHODS: A model of biliary injury in dogs was made and then a (103)Pd radioactive stent was positioned in the biliary duct. The expression and distribution of SMA were assessed in the anastomotic tissue 30 days after implantation of the stent. RESULTS: SMA expression was less in the (103)Pd stent group than in the ordinary stent group. The (103)Pd stent inhibited scar contracture and anastomotic stenosis. CONCLUSION: The (103)Pd stent can reduce the expression of SMA in the healing process and inhibit scar contracture and anastomotic stenosis in the dog biliary duct.

Is delayed gastric emptying so terrible after pylorus-preserving pancreaticoduodenectomy? Prevention and management.

AIM: To explore some operative techniques to prevent the occurrence of delayed gastric emptying (DGE) after pylorus-preserving pancreaticoduodenectomy (PPPD). METHODS: One hundred and eighty-six patients in a single medical center who accepted PPPD were retrospectively studied. The incidence of DGE was investigated and the influence of some operative techniques on the prevention of DGE was analyzed. RESULTS: During the operative process of PPPD, the methods of detached drainage of pancreatic fluid and bile and gastric fistulization were used. Postoperatively, six patients suffered DGE among the 186 cases; the incidence was 3.23% (6/186). One of them was complicated with intraabdominal infection at the same time, and two with pancreatic leakage. CONCLUSION: Appropriate maneuvers during operation are essential to avoid postoperative DGE in PPPD. The occurrence of DGE is avoidable. It should not be used as an argument to advocate hemigastrectomy in PPPD.

[Expression and significance of caspase-3 gene in apoptotic muscle cells 103Pd radioactive stent bile duct in dogs].

OBJECTIVE: To discuss the expression and significance of caspase-3 gene in the apoptotic muscle cells in gamma-radiation-induced muscle cell lines. METHODS: The caspase-3 mRNA in the control and gamma-radiation induced apoptotic muscle cells was analysed by RT-PCR. RESULTS: The expression of caspase-3 gene transcript was higher in 103Pd radioactive stent dog bile duct than in general stent dog bile duct, and apoptotic muscle cells were higher in 103Pd radioactive stent dog bile duct than in general stent dog bile duct. CONCLUSIONS: The high level expression of caspase-3 gene may help to understand the muscle cells sensitivity to gamma-radiation apoptosis. 103Pd radioactive stent may increase the expression of caspase-3 gene in dog bile duct and prevent the billiary narrow when dog bile duct was injured by balloon.

[Effect of 103palladium radioactive stent on expression of smooth muscle actin in bile duct during healing process and its significance].

OBJECTIVE: To observe the effect of radiation on the expression of smooth muscle actin (SMA) in the bile duct during the healing process and the inhibitory function of (103)palladium (Pd) radioactive stent on the stricture of bile duct after injury. METHODS: Twelve mongrel dogs were made models of bile duct injury: duodenotomy was performed, a balloon catheter was inserted into the general bile duct and saline with high pressure was perfused thereinto to cause laceration of the mucosa, and then the balloon catheter was withdrawn and ordinary alloy stent or (103)Pd radioactive stent was inserted into the general bile duct. Thirty days after the dogs were killed. Their bile ducts were taken out to undergo HE staining to observe the area of general bile duct, thickness of the tunica intima, area of residual bile duct cavity, stricture degree, and circumference of bile duct. The expression of SMA in the bile duct tissue was detected by immunoistochemistry. RESULTS: SMA was expressed in 5 of the 6 specimens of bile duct in the (103)Pd radioactive stent group and 2 of the 6 specimens of the ordinary stent group (P < 0.01). The maximum thickness of tunica intima of general bile duct was 0.78 mm +/- 0.12 mm in the (103)Pd radioactive stent group, significantly less than that of the ordinary stent group (1.86 mm +/- 0.14 mm, P < 0.01). The percentage of maximum stricture area of the (103)Pd radioactive stent group was 23% +/- 16%, significantly lower that that of the ordinary stent group (56% +/- 22%, P < 0.01). The circumference of bile duct cavity of the (103)Pd radioactive stent group was 9.7 mm +/- 1.6 mm, significantly longer that of the ordinary stent group (7.0 mm +/- 1.4 mm, P < 0.01). CONCLUSION: (103)Pd radioactive stent reduces the expression of SMA in the bile duct during the healing process, thus inhibiting the stricture of bile duct caused by scar contracture at the anastomotic stoma.

Attenuation of small-for-size liver graft injury by FTY720: significance of cell-survival Akt signaling pathway.

To investigate the protective mechanism of FTY720 in small-for-size liver grafts, we applied a rat orthotopic liver transplantation model using 40% of liver grafts. FTY720 was administered (1 mg/kg, i.v.) at 20 min before graft harvesting in the donor, immediately before total hepatectomy and immediately after graft reperfusion in the recipient. The 7-day graft survival rates in the FTY720 group were significantly improved compared with the control group [100% (6/6) vs. 40% (4/10), p = 0.034]. FTY720 significantly reduced serum ALT and AST levels at 24 h after liver transplantation. The cell-survival Akt signaling pathway was activated in FTY720 groups by phosphorylation of Glycogen Synthase Kinase-3beta, Bad and Forkhead Transcription Factor at 6 and 24 h after liver transplantation. The cleaved-caspases 3, 7 and 9 were down-regulated, accompanied with less apoptotic nuclei after FTY720 treatment. Acute-phase inflammatory MAPK pathway was down-regulated by dephosphorylation of c-Raf, Mek and Erk in the treatment groups. A20 and endothelial nitric oxide synthase were up-regulated together with down-regulation of iNOS. Hepatic sinusoids were well preserved in the FTY720 group but disrupted in the control group. In conclusion, FTY720 attenuates small-for-size liver graft injury by activation of cell-survival Akt signaling and down-regulation of the MAPK pathway.

Intrabiliary radiation inhibits smooth muscle formation and biliary duct remodelling after balloon overstretching injury in dogs.

BACKGROUND: Internal metallic stents have been widely used in clinical practice, but a high postoperative restenosis rate limits its application. The purpose of this study was to determine the effect of intrabiliary radiation on muscle formation and biliary duct remodeling after biliary duct balloon injury in dogs. METHODS: Twenty male dogs (15 - 20 kg) were randomly divided into treatment group (n = 10) and control group (n = 10). Balloon overstretching injury was induced using a balloon catheter placed across the biliary duct. Subsequently, a 103Pd radioactive stent was positioned at the target site in each animal in the treatment group, providing the injured biliary duct with a radiation dose of 12.58 x 10(7) Bq. Dogs in the control group received Ni-Ti stents. All the dogs were killed one month after initial injury. The injured sections were dissected free from the dogs, and were processed for histological and morphological study. Cross-sections were stained with hematoxylin-eosin, Masson's trichrome, and Verhoef-van Giesen. Muscle formation area and lumen area were determined using a computer-assisted image analysis system. RESULTS: Compared with the control group, 103Pd radioactive stents significantly reduced muscle formation area (78.3%, P < 0.01), and percentage area of stenosis [control stents: (60.0 +/- 21.6)%, 103Pd radioactive stents: (31.6 +/- 9.5)%]. In addition, in the treatment group, the biliary duct lumen area was significantly larger than that in the control group (P < 0.01). CONCLUSIONS: 103Pd radioactive stents providing a radioactive dose of 12.58 x 10(7) Bq are effective in reducing muscle formation and biliary duct remodeling after balloon overstretching injury.

Prevention of pancreaticojejunal anastomotic leakage after pancreaticoduodenectomy with separate internal drainage of bile and pancreatic fluid.

OBJECTIVE: To introduce a new reconstructional procedure to decrease the complications after pancreaticoduodenectomy. METHODS: Separate internal drainage of bile and pancreatic fluid in pancreaticoduodenectomy was performed in 256 patients. The digestive tract was reconstructed with Child method, with invaginated pancreaticojejunostomy using a long silastic tube to drain pancreatic fluid internally, an end-to-side choledochojejunostomy and an end-to-side duodenojejunostomy or gastrojejunostomy. Gastrostomy drainage was also performed. RESULTS: No complications of pancreatic leakage were found. CONCLUSION: The separate internal drainage of bile and pancreatic fluid plays an important role in preventing pancreaticojejunal anastomotic leakage.

Iatrogenic extrahepatic bile duct injury in 182 patients: causes and management.

OBJECTIVE: To describe the causes and treatment of iatrogenic bile duct injury caused by cholecystectomy. METHODS: 182 patients with iatrogenic extrahepatic bile duct injury from 4 university hospitals of China were reviewed. Details of primary cholecystectomy, biliary reconstruction as well as postoperative management were recorded. All patients were followed up for at least 6 months (6 months to 9 years, median 3.5 years). The adequacy of repair was assessed by regular evaluation of the patients' clinical status and liver function variables. Hepatobiliary B-ultrasonography was used routinely in the follow up of patients, and magnetic resonance cholangiopancreatography was applied in the patients suggestive of abnormality. RESULTS: In 152 patients, bile duct injury happened during open cholecystectomy, and in 30 patients during laparoscopic cholecystectomy. All the injuries developed during anterograde cholecystectomy (at the Calot's triangle). All the patients with these injuries underwent choledochocholedochostomy or Roux-en-Y choledochojejunostomy with good results (161 patients), recurrent stricture (11), and death (10). CONCLUSIONS: During cholecystectomy, the Calot's triangle should be identified anatomically, but retrograde cholecystectomy is the optimal choice. Bile duct injury should be discovered as soon as possible and be managed timely. Different operative methods are optional according to the degree of injury and the postoperative period.