Hydrogen-rich saline protects against ischemia/reperfusion injury in grafts after pancreas transplantations by reducing oxidative stress in rats.

PURPOSE: This study aimed to investigate the therapeutic potential of hydrogen-rich saline on pancreatic ischemia/reperfusion (I/R) injury in rats. METHODS: Eighty heterotopic pancreas transplantations (HPT) were performed in syngenic rats. The receptors were randomized blindly into the following three groups: the HPT group and two groups that underwent transplantation and administration of hydrogen-rich saline (HS, >0.6 mM, 6 mL/kg) or normal saline (NS, 6 mL/kg) via the tail vein at the beginning of reperfusion (HPT + HS group, HPT + NS group). Samples from the pancreas and blood were taken at 12 hours after reperfusion. The protective effects of hydrogen-rich saline against I/R injury were evaluated by determining the changes in histopathology and measuring serological parameters, oxidative stress-associated molecules, and proinflammatory cytokines. RESULTS: Administration of hydrogen-rich saline produced notable protection against pancreatic I/R injury in rats. Histopathological improvements and recovery of impaired pancreatic function were observed. In addition, TNF-α, IL-1ß, and IL-6 were reduced markedly in the HPT + HS group. Additionally, there were noticeable inhibitory effects on the pancreatic malondialdehyde level and considerable recruitment of SOD and GPx, which are antioxidants. CONCLUSION: Hydrogen-rich saline treatment significantly attenuated the severity of pancreatic I/R injury in rats, possibly by reducing oxidative stress and inflammation.

Involvement of a membrane potassium channel in heparan sulphate-induced activation of macrophages.

Increasing evidence has demonstrated that Toll-like receptor 4 (TLR4) -mediated systemic inflammatory response syndrome accompanied by multiple organ failure, is one of the most common causes of death in patients with severe acute pancreatitis. Recent reports have revealed that heparan sulphate (HS) proteoglycan, a component of extracellular matrices, potentiates the activation of intracellular pro-inflammatory responses via TLR4, contributing to the aggravation of acute pancreatitis. However, little is known about the participants in the HS/TLR4-mediated inflammatory cascades. Our previous work provided a clue that a membrane potassium channel (MaxiK) is responsible for HS-induced production of inflammatory cytokines. Therefore, in this report we attempted to reveal the roles of MaxiK in the activation of macrophages stimulated by HS. Our results showed that incubation of RAW264.7 cells with HS up-regulated MaxiK and TLR4 expression levels. HS could also activate MaxiK channels to promote the efflux of potassium ions from cells, as measured by the elevated activity of caspase-1, whereas this was significantly abolished by treatment with paxilline, a specific blocker of the MaxiK channel. Moreover, it was found that paxilline substantially inhibited HS-induced activation of several different transcription factors in macrophages, including nuclear factor-κB, p38 and interferon regulatory factor-3, followed by decreased production of tumour necrosis factor-α and interferon-ß. Taken together, our investigation provides evidence that the HS/TLR4-mediated intracellular inflammatory cascade depends on the activation of MaxiK, which may offer an important opportunity for a new approach in therapeutic strategies of severe acute pancreatitis.

Effects of treatment with hydrogen sulfide on methionine-choline deficient diet-induced non-alcoholic steatohepatitis in rats.

BACKGROUND AND AIM: Oxidative stress and inflammation play important roles in the progression from simple fatty liver to non-alcoholic steatohepatitis (NASH). The aim of this work was to investigate whether treatment with hydrogen sulfide (H2 S) prevented NASH in rats through abating oxidative stress and suppressing inflammation. METHODS: A methionine-choline-deficient (MCD) diet rat model was prepared. Rats were divided into three experimental groups and fed for 8 weeks as follows: (i) control rats; (ii) MCD-diet-fed rats; (iii) MCD-diet-fed rats treated with NaHS (intraperitoneal injection of 0.1 mL/kg/day of 0.28 mol/L NaHS, a donor of H2 S). RESULTS: MCD diet impaired hepatic H2 S biosynthesis in rats. Treatment with H2 S prevented MCD-diet-induced NASH, as evidenced by hematoxylin and eosin staining, reduced apoptosis and activities of alanine aminotransferase and aspartate aminotransferase, and attenuated hepatic fat accumulation in rats. Treatment with H2 S abated MCD-diet-induced oxidative stress through reducing cytochrome p4502E1 expression, enhancing heme oxygenase-1 expression, and suppressing mitochondrial reactive oxygen species formation, and suppressed MCD-diet-induced inflammation through suppressing activated nuclear factor κB signaling and reducing interleukin-6 and tumor necrosis factor α expressions. In addition, treatment of MCD-diet fed rats with H2 S had a beneficial modulation on expression profiles of fatty acid metabolism genes in livers. CONCLUSIONS: Treatment with H2 S prevented NASH induced by MCD diet in rats possibly through abating oxidative stress and suppressing inflammation.

Phospholipase A2 enzymes PLA2G2A and PLA2G12B as potential diagnostic and prognostic biomarkers in cholangiocarcinoma.

BACKGROUND: Phospholipase A2 (PLA2) enzymes are pivotal in various biological processes, such as lipid mediator production, membrane remodeling, bioenergetics, and maintaining the body surface barrier. Notably, these enzymes play a significant role in the development of diverse tumors. AIM: To systematically and comprehensively explore the expression of the PLA2 family genes and their potential implications in cholangiocarcinoma (CCA). METHODS: We conducted an analysis of five CCA datasets from The Cancer Genome Atlas and the Gene Expression Omnibus. The study identified differentially expressed genes between tumor tissues and adjacent normal tissues, with a focus on PLA2G2A and PLA2G12B. Gene Set Enrichment Analysis was utilized to pinpoint associated pathways. Moreover, relevant hub genes and microRNAs for PLA2G2A and PLA2G12B were predicted, and their correlation with the prognosis of CCA was evaluated. RESULTS: PLA2G2A and PLA2G12B were discerned as differentially expressed in CCA, manifesting significant variations in expression levels in urine and serum between CCA patients and healthy individuals. Elevated expression of PLA2G2A was correlated with poorer overall survival in CCA patients. Additionally, the study delineated pathways and miRNAs associated with these genes. CONCLUSION: Our findings suggest that PLA2G2A and PLA2G12B may serve as novel potential diagnostic and prognostic markers for CCA. The increased levels of these genes in biological fluids could be employed as non-invasive markers for CCA, and their expression levels are indicative of prognosis, underscoring their potential utility in clinical settings.

Progressive balloon dilatation following hepaticojejunostomy improves outcome of bile duct stricture after iatrogenic biliary injury.

BACKGROUND: Iatrogenic biliary stricture (IBS) is a disastrous complication of cholecystectomy. Although the endoscopic treatments are well accepted as initial attempts for IBS, surgical hepaticojejunostomy (HJ) is often necessary for a considerable proportion of patients. However, the anastomotic stricture after HJ also occurs. METHODS: In the present study, a new procedure, progressive balloon dilation following HJ (HJPBD), was designed and utilized in the IBS treatment. We retrospectively compared HJPBD with the traditional HJ in term of the outcomes when used for IBS treatment. RESULTS: Between January 1997 and December 2009, 112 patients with IBS attributed to cholecystectomy enrolled in our hospital were treated with surgical reconstruction with either HJ (n=58) or HJPBD (n=54). Of the 58 patients in HJ group, 48 patients (82.8%) had a successful outcome, while 52 out of 54 patients (96.3%) in HJPBD group achieved success. The successful surgical reconstruction rates were significantly different between these two groups, with a further improved outcome in patient undergone progressive balloon dilation following HJ. Additionally, 8 of the 10 failure cases in HJ group were successfully rescued by HJPBD procedure. CONCLUSIONS: Our findings suggest that the new procedure of HJPBD could be successfully applied to IBS patients, and significantly improve the outcome of IBS reconstruction.

Abdominal paracentesis drainage improves outcome of acute pancreatitis complicated with intra-abdominal hypertension in early phase.

BACKGROUND: Intra-abdominal hypertension (IAH) is an important risk factor for organ dysfunction, and it occurs in the early phase of severe acute pancreatitis (SAP). We have reported a novel step-up approach and shown the benefit of performing abdominal paracentesis drainage (APD) ahead of percutaneous catheter drainage (PCD) when treating Patients with SAP with fluid collections. This study aimed to evaluate the efficacy of APD in Patients with SAP complicated with IAH in the early phase. METHODS: In the present study, 206 AP patients complicated with IAH in the early phase were enrolled in hospital between June 2017 and December 2020. The patients were divided into two groups: 109 underwent APD (APD group) and 97 were managed without APD (non-APD group). We retrospectively compared the outcomes of the APD and non-APD groups for IAH treatment. The parameters including mortality, infection, organ failure, inflammatory factors, indications for further interventions, and drainage-related complications were observed. RESULTS: The demographic data and severity scores of the two groups were comparable. The mortality rate was lower in the APD group (3.7%) than in the non-APD group (8.2%). Compared with the non-APD group, the intra-abdominal pressure and laboratory parameters of the APD group decreased more rapidly, and the mean number of failed organs was lower. However, there was no significant difference in incidence of infections between the two groups. CONCLUSIONS: Application of APD is beneficial to AP patients. It significantly attenuated inflammation injury, avoided further interventions, and reduced multiple organ failure.

[Protective effect of Ligustrazine preconditioning on hepatic energy metabolism in ischemia/reperfusion injury of rat].

OBJECTIVE: Objective to investigate the protective effects of Ligustrazine preconditioning against hepatic ischemia/reperfusion injury (IRI) in rats. METHODS: Fifty male Wistar rats were randomly allocated into 3 groups: sham operation group, in which animals underwent laparotomy, experimental group and control group in which were treated with 70% IRI of the liver, especially, the animals in experimental group was given intraperitoneal injection of 2 mL Ligustrazine per day for 3 days before operation. After the operation, liver tissues were harvested at 1 h, 6 h, 24 h and 72 h for the study of histomorphological change, the respiratory control ratio (RCR) and phosphorus: oxygen ratio (P/O) of hepatocytes mitochondria, and the contents of adenosine triphosphate (ATP) of liver tissue. RESULTS: (1) The damage hepatic tissue in experimental group was slighter than that in control group at each corresponding time-point after operation. (2) The RCR and P/O ratio at each corresponding time-point were higher in experimental group than those in control group (P < 0.05), and all of the two groups recovered after 72 h. (3) The ATP concentration in experimental group also was higher than that in control group at each corresponding time-point, and recovered faster than control group. CONCLUSION: The current results show that Ligustrazine preconditioning may improve energy metabolism of rat liver in ischemia reperfusion injury.

[Protective effect of pre-storing glycogen on warm ischemia reperfusion injury in rat liver].

OBJECTIVE: To study the effect of pre-storing glycogen on warm ischemia reperfusion injury in rat liver. METHODS: Lewis rats were divided into sham operation group (S group), low-glycogen group (L group, fasted 24 h), normal-glycogen group (N group, standard laboratory diet), high-glycogen group (H group, standard laboratory diet plus intravenous injection of 50% glucose solution 1 mL every 6 h x 4 times). Seventy percent portal ligation was performed on all of the rats for 30 min except for those in the S group. Ten rats from each group were sacrificed for harvest of serum and tissue samples 1 h, 4 h, 12 h and 24 h after reperfusion, respectively. The hepatic function was measured and the morphological changes of the livers were examined. Bcl-2, a well known antiapoptotic factor, was also detected using quantitative polymerase chain reaction. RESULTS: The rats with higher glucose presented higher glycogen in hepatocytes, better hepatic function, lower levels of ALT, AST and apoptosis index (AI), and higher 1 week survival rate. These rats also showed slighter histological damage and lower apoptotic index than the rats in the other groups. Furthermore, the morphological changes of the liver tissues of the rats with higher glucose were mild. The Bcl-2 mRNA expression was the strongest. CONCLUSION: Pre-storing glycogen might protect liver impairment caused by ischemia reperfusion injury, perhaps through enhancing the Bcl-2 expression and preventing the hepatic cells from apoptosis.

Comparatively lower postoperative hepatolithiasis risk with hepaticocholedochostomy versus hepaticojejunostomy.

BACKGROUND: Optimal surgical technique plays a key role in preventing the postoperative recurrence of hepatolithiasis. Tian et al developed the subcutaneous tunnel and hepatocholangioplasty using the gallbladder (STHG) technique and applied it in hepatolithiasis patients who had an approximately normal gallbladder and sphincter of Oddi. However, the technique is controversial. In the present study, a canine model was established for hepatocholangioplasty (HC) and hepaticojejunostomy (HJ) to simulate STHG and Roux-en-Y cholangiojejunostomy in the clinic, respectively. Then, the alterations of bile components in the vicinity of the anastomosis were compared. This may provide an experimental guide for choosing an optimal technique for the treatment of hepatolithiasis in the clinic. METHODS: The animals were randomly separated into a control group (5 dogs) and a model group (stenosis of the common bile duct; 24 dogs). The 24 dogs in the model group were randomly divided into an HC group and an HJ group (12/group). Bile was collected from the bile duct at 1 and 5 months after the operation, and the bile components were determined. RESULTS: The levels of total bile acid, cholesterol, total bilirubin, and phospholipid in the HC group were higher than those in the HJ group (P<0.05). However, no statistical difference was seen in unconjugated bilirubin and calcium ions. The mucin level in bile in the HC group was lower than that in the HJ group at 5 months after the operation (P<0.05). The postoperative lipid peroxidation level was remarkably lower than that in the HJ group (P<0.05). However, the superoxide dismutase level was remarkably higher than that in the HJ group (P<0.05). Finally, a significant difference was found in the positive bacterial culture rate in bile between the groups. CONCLUSION: Changes of bile components near the anastomosis after HC might be more preferable for preventing hepatolithiasis formation than HJ.

[Effect of 90 percentage portal branch ligation on liver regeneration and expression of metalloproteinase and tissue inhibitors of metalloproteinases in rats].

OBJECTIVE: To investigate the effect of 90% portal branch ligation on liver regeneration and expression of metalloproteinases and tissue inhibitors of metalloproteinases in rats. METHODS: Ninety-six SD rats were randomly divided into Sham-PBL group and portal vein branches ligation group. The weight of both ligated and unligated lobes of liver were measured at post operation day (POD) 0.5, 1, 3, 5, 7, 14, 21 and 28. The morphological changes of the non-ligated liver lobes were observed by microscope. The expression of PCNA, MMP2, MMP9 and TIMP2 of the non-ligated liver lobes were studied by immunohistochemistry. RESULTS: 1) 95.8% rats survived from the ligation of 90% portal branch. Hepatic lobe at the ligated side diminished progressively after ligation, whereas the lobes of the unligated side underwent compensatory regeneration. The ratio of non-ligated lobes weight to the whole liver increased slowly within 1d, speeded up significantly during 1-5d period, increased slowly after POD5, and got the plateau stag at POD7; 2) PCNA index were markedly increased within POD 0.5-3 (P < 0.01). It reached the peak at POD5 and decreased slightly at POD7, but still higher than Sham-PBL group level, then gradually returned to normal. 3) The expression of MMP2,MMP9 and TIMP2 in the non-ligated liver lobes were markedly increased at 1d. It reached the peak at POD7 and gradually returned to normal within POD7-28. 4) The MMP2 and PCNA in liver had a positive correlation at POD 0.5, 1, 5, 7, 14. The expressions of MMP9 and PCNA had a positive correlation at POD 0.5, 1, 7, 21. CONCLUSION: The expressions of TIMP2 and PCNA had a positive correlation at POD1, 7, 14, 21. The expression of MMP2, MMP9 and TIMP2 of the non-ligated liver lobes is markedly increased at POD1. It reaches the peak at POD7, and dropped to normal level gradually. The expressions of MMP2, MMP9 and TIMP2 and PCNA were correlated in 90% portal branch Ligation rats. The expression of MMP2,MMP9 and TIMP2 may play a pivotal role in liver regeneration.

Bile Acid Transporters Are Expressed and Heterogeneously Distributed in Rat Bile Ducts.

Background/Aims: Cholangiocytes are capable of reabsorbing bile salts from bile, but the pathophysiological significance of this process is unclear. To this end, we detected the expression and distribution of bile acid transport proteins in cholangiocytes from normal rat liver and analyzed the possible pathophysiological significance. Methods: Bile duct tissues of Sprague-Dawley rats were isolated by enzymatic digestion and mechanical isolation, and then divided into large and small bile duct tissues. Immunohistochemistry, real-time polymerase chain reaction and Western blotting were used to determine the expression of the apical sodium-dependent bile acid transporter (ASBT), ileal bile acid binding protein (IBABP), and basolateral organic solute transporter α (Ostα) in the biliary tract system of rats. Differences in the expression and distribution of these proteins were analyzed. Results: In cholangiocytes, ASBT and IBABP were mainly expressed in cholangiocytes of the large bile ducts, in which the expression of both was significantly higher than that in the small ducts (p<0.05). Ostα was simultaneously expressed in cholangiocytes of both the large and small bile ducts, showing no significant difference in expression between the two groups of bile ducts (p>0.05). Conclusions: Bile acid transporters are expressed and heterogeneously distributed in rat bile ducts, indicating that bile acid reabsorption by cholangiocytes might mainly occur in the large bile ducts. These findings may help explore the physiology of bile ducts and the pathogenesis of various cholangiopathies.

[Hepatocellular glycogen alleviates hepatic ischemia reperfusion injury and its relationship to ICAM-1 gene expression].

OBJECTIVE: To investigate if higher hepatocellular glycogen contents can alleviate hepatic ischemia reperfusion injury and its relationship to ICAM-1 gene expression in hepatic sinusoidal cells (HSCs). METHODS: Twenty-one rabbits fed with a standard diet were randomly divided into three groups (n=7 in each). All the animals were subjected to hepatic ischemia reperfusion injury then sacrificed. Before the injury, group A rabbits fasted for 24 hours; group C rabbits had 6 intravenous glucose solution (25%, 20 ml) injections, 4 hours between two injections. Hepatic enzymological changes, hepatic ICAM-1 mRNA expressions and leukocytic counts in the sinusoids were observed. RESULTS: The liver glycogen contents of the three groups were significantly different. Livers of group A had higher contents of glycogen (9.85+/-0.91 mg/g. wet tissue); in group B they were 38.93+/-5.72; and in group C they were 48.31+/-6.58. Group C animals had the slightest liver function damage. There were no differences in the pre- and post-ischemic ICAM-1 mRNA contents in the three groups. However, livers with a higher content of glycogen showed less expression of ICAM-1 mRNA (group A: 1.398+/-0.365 ng/mg wet tissue; group B: 0.852+/-0.297; group C: 0.366+/-0.183) and lower leukocytic counts. The relationship analysis showed a negative relationship between hepatocellular glycogen and hepatic ICAM-1 mRNA contents (r= -0.965, P less than 0.01). CONCLUSIONS: Hepatocellular glycogen is important in protecting liver ischemic reperfusion injury. Also hepatocellular glycogen decreases the expression of ICAM-1 mRNA of HSCs.

[Ultrasound-guided percutaneous tube drainage combined with directly-viewed debridement with cholangioscopy: a mini-invasive strategy for peripancreatic necrotizing infection].

OBJECTIVE: To establish "an integrative therapy" of drainage and debridement on peripancreatic necrotizing infection (PPNI) with minimally invasive technique, and to detect its clinical effects. METHODS: There were 17 patients who accepted ultrasound-guided percutaneous tube drainage combined with directly-viewed debridement with cholangioscopy from March 2006 to January 2008. Percutaneous puncture and catheter (6 - 8 F) drainage were adopted on the patients suffering from PPNI with B-us guidance, then the drainage sinus was expanded progressively from 8 F to 24 F in diameter with Cook fascia dilator by degrees, and the 22 F or 24 F tube was easily placed into the interior of PPNI instead of the prior catheter. So a better drainage effect was achieved. One week later, the necrotizing tissue of PPNI could be observed and debrided with choledochoscope under a directly-viewed way through the enlarged new sinus. Thus, with the continuous tube drainage and repeated debridement, the focus was absorbed and covered gradually. RESULTS: Seventeen cases accepted the mini-invasive therapy, 15 cases were saved finally with cure rate of 88.2%, and 2 cases conversion to laparotomy because of some technical reasons. The mean healing time was 73 days, and the mean hospitalization time was 57 days. Bleeding was occurred in 2 cases localized in sinus and the inside of PPNI, digestive tract fistula was detected in 2 cases, and these patients with the complications were cured under nonoperative management. All the patients were still alive with following-up, neither remains nor recurrence of the PPNI was found in our group. CONCLUSIONS: Ultrasound-guided percutaneous tube drainage combined with directly-viewed debridement with cholangioscopy, as a mini-invasive therapy, could complete the goal-directed therapy of PPNI, meanwhile, realize the modern surgery ideal of damage control.

Long Noncoding RNA NEAT1 Promotes Growth and Metastasis of Cholangiocarcinoma Cells.

Long noncoding RNAs (lncRNAs) are known to play important roles in cancers. However, little is known about lncRNAs in cholangiocarcinoma (CCA), a cholangiocyte malignancy with poor prognosis. We investigated the role of nuclear paraspeckle assembly transcript 1 (NEAT1) lncRNA in promoting CCA. qRT-PCR analysis of patient samples showed that NEAT1 expression was higher in CCA tumors than in matched adjacent nontumor tissue. NEAT1 levels were also higher in CCA cell lines than in a normal biliary epithelium cell line (HIBEpic). NEAT1 knockdown in CCA cell lines using shNEAT1 reduced cell proliferation and colony formation in CCK-8 and colony formation assays, respectively. CCA cells transfected with shNEAT1 also exhibited reduced metastasis and invasiveness in Transwell assays. NEAT1 knockdown cells produced smaller tumors, demonstrating that NEAT1 promotes tumor growth in vivo. Silencing of NEAT1 increased E-cadherin expression in vitro, and E-cadherin expression was inversely correlated with NEAT1 expression in CCA tissue samples. RIP and ChIP assays suggest that NEAT1 is recruited to the E-cadherin promoter by EZH2 (enhancer of zeste homolog 2), where it represses E-cadherin expression. These findings indicate that NEAT1 exerts oncogenic effects in CCA. We postulate that NEAT1 is a potentially useful diagnostic and therapeutic target for CCA.

Ultrasonic interventional analgesia in pancreatic carcinoma with chemical destruction of celiac ganglion.

AIM: To detect the therapeutic effects of chemical destruction of celiac ganglion in patients with pancreatic carcinoma with intractable pain. METHODS: Ninety-seven cases with advanced pancreatic carcinoma received chemical destruction of celiac ganglion-5 mL pure alcohol injection around celiac artery under ultrasonic guidance. The changes of visual analogue scale (VAS), serum substance P (Sub P), beta-endopeptide (beta-EP) and T-lymphocyte subtypes level were compared between pre- and post-therapy. RESULTS: Successful rate of puncture was 98.7%, with one failure. No serious complications such as traumatic pancreatitis, pancreatic fistula, abdominal cavity hemorrhage or peritoneal infection occurred. VAS, serum Sub P and beta-EP level significantly changed after treatment (8.0+/-2.3 vs 4.6+/-2.1, 254.1+/-96.7 vs 182.4+/-77.6, 3.2+/-0.8 vs 8.8+/-2.1, P<0.01, P<0.05, P<0.01) with complete relief rate 54.2%, partial relief rate 21.9%, ineffective rate 12.5% and recurrent rate 10.7%. The T-lymphocyte subtypes level remarkably increased when compared with that of pre-therapy (46.7+/-3.7 vs 62.5+/-5.5, P<0.01). CONCLUSION: Our study suggests that chemical destruction of celiac ganglion under ultrasonic guidance is highly safe, and can evidently relieve cancer pain and improve the cellular immunity in patients with advanced pancreatic carcinoma.

[Perspective of pre-operational jaundice-reducing indication in carcinoma of head of pancreas].

OBJECTIVE: To explore the necessity, advantages and disadvantages of reducing the Icterus Index before operation in carcinoma of the head of pancreas. METHODS: A total of 183 patients with serum total bilirubin (TB) level higher than 220 micromol/L were randomized into 2 groups: jaundice-reducing group (92 patients) and non-reducing group (91 patients). In jaundice-reducing group, all the patients were performed ultrasound-guided percutaneous transhepatic bile duct drainage (UPTBD) and endoscopic nasobiliary drainage (ENBD). The jaundice-reducing group was operated on 3 weeks after tube placement. In non-reducing group, all the patients underwent operations only after general pre-operation routine preparation within 5 days after admission. The operation and post-operation recovery in the two groups was investigated and compared. RESULTS: In jaundice-reducing group, the level of TB decreased to 120 micromol/L from 279 micromol/L in 89 patients after biliary drainage. Of the 89 patients, pancreatoduodenectomy was successfully performed in 39 (43.8%), 47 (52.8%) underwent simple internal drainage and the other 3 were just explored. The average blood loss was 250 ml (110 - 980 ml), complications were found in 8 patients (9.0%) and one died. In non-reducing group, pancreatoduodenectomy was successfully performed in 24 (26.4%), simple internal drainage in 58 patients (63.7%) and exploration in 9 (9.9%). The average blood loss was 480 ml (320 - 1750 ml), complications were found in 19 patients (20.9%) and 4 died. In the non-reducing group, the patients with complications were older than those without complications, and the TB level was higher. The excision rate of carcinoma, incidence rate of complications and hospital time in patients whose TB decreased over 30% weekly after reducing the Icterus Index were all better than those of the rest. CONCLUSIONS: It is necessary to reduce the Icterus Index before operation in the patients with carcinoma of head of pancreas complicated with serious jaundice, especially for the elder, which can not only reduce the blood loss but also make operations safer and increase cure rate, in addition. And whether the Icterus Index decreases smoothly with biliary drainage can be used to predict the operational risk, effect and prognosis.

Negative correlation of ITCH E3 ubiquitin ligase and miRNA-106b dictates metastatic progression in pancreatic cancer.

Pancreatic cancer is one of the major malignancies and cause for mortality across the world, with recurrence and metastatic progression remaining the single largest cause of pancreatic cancer mortality. Hence it is imperative to develop novel biomarkers of pancreatic cancer prognosis. The E3 ubiquitin ligase ITCH has been previously reported to inhibit the tumor suppressive Hippo signaling by suppressing LATS1/2 in breast cancer and chronic lymphocytic leukemia. However, the role of ITCH in pancreatic cancer progression has not been described. Here we report that ITCH transcript and protein expression mimic metastatic trait in pancreatic cancer patients and cell lines. Loss-of-function studies of ITCH showed that the gene product is responsible for inducing metastasis in vivo. We furthermore show that hsa-miR-106b, which itself is down regulated in metastatic pancreatic cancer, directly interacts and inhibit ITCH expression. ITCH and hsa-miR-106b are thus potential biomarkers for pancreatic cancer prognosis.

Middle and long-term clinical outcomes of patients with regional hepatolithiasis after subcutaneous tunnel and hepatocholangioplasty with utilization of the gallbladder.

BACKGROUND: Roux-en-Y choledochojejunostomy is routinely performed in patients with regional hepatolithiasis. However, some of these patients, who have a normal gallbladder and normal Oddi's sphincter, are unnecessarily undergoing bilio-intestinal drainage. Alternatively, reconstruction can be achieved by subcutaneous tunnel and hepatocholangioplasty with the utilization of the gallbladder (STHG). This method is effective to potential endoscopic tunnel and intervention during follow-up, and prevention of reflux cholangitis as well as the disorders of the GI tract. METHODS: The middle and long-term complications of 46 patients who underwent STHG were analyzed. With B-ultrasonography and biochemical assay, the contraction and concentration function of the gallbladder were also studied. RESULTS: Follow-up showed that all patients survived with a relatively normal life. One patient experienced right epigastric pain, chills and fever because of a stone which impacted in the left hepatic bile duct. Another patient had cholangitis because of biliary ascariasis. The two patients were treated by endoscopic therapy within the subcutaneous gallbladder under local anesthesia. CONCLUSIONS: This operation not only keeps the normal physical functional of the gallbladder, Oddi's sphincter and gastrointestinal tract, but also prevents reflux cholangitis and the disorder of the digestive tract. Hence STHG is a novel operation dealing with regional hepatolithiasis.

[Construction of GFP-AWP1 fusion gene vector and its expression in 293 cells].

OBJECTIVE: To construct green fluorescent protein (GFP)-AWP1 (a novel human protein associated with protein kinase C-related kinase 1) fusion gene vector for observing the expression and localization of AWP1 in 293 cells. METHODS: The coding region in AWP1 cDNA was amplified by RT-PCR from human endothelial cell line ECV304 and recombined into pEGFP-C2 plasmid expressing GFP. After identification with restriction endonucleases and sequence analysis, the recombinant plasmid was transfected into 293 cells with the cationic liposome DOTAP as the transfection reagent. The expression and localization of AWP1 were observed under a fluorescence microscope. RESULTS: Restriction endonuclease assay and sequence analysis verified the successful construction of the recombinant vector pEGFP-C2/AWP1, and GFP-AWP1 fusion protein was highly efficiently expressed in 293 cells. Under fluorescent microscope, green fluorescence was seen homogeneously distributed in the entire cell body of the cells transfected by the empty vector pEGFP-C2, but diffusely in the cytoplasm of the cells transfected by the recombinant vector pEGFP-C2/AWP1. CONCLUSION: GFP-AWP1 fusion gene vector is successfully constructed and the fusion protein expressed in the cytoplasm of 293 cells.

Enhancing HOTAIR/MiR-10b Drives Normal Liver Stem Cells Toward a Tendency to Malignant Transformation Through Inducing Epithelial- to-Mesenchymal Transition.

Previously, other groups and our team consistently have demonstrated that the possible origination of liver cancer stem cells (LCSCs) is the malignant transformation from liver normal stem cells (LNSCs). However, this complex and multi-step process is far from clear due to the accumulation of various gene dysregulations. Because non-coding RNAs (ncRNAs) could regulate multiple genes, a family of genes, and even whole chromosomes, this study further investigated the effect of dysregulated short ncRNA microRNA-10b and long ncRNA HOX transcript antisense RNA (HOTAIR) between LNSCs and LCSCs on phenotype reversion. To clarify the role of ncRNA in malignant transformation of LNSCs, we used lentivirus transduction to enhance the miR-10b and HOTAIR expression levels in our previously isolated rat LNSCs. The malignant abilities of proliferation, invasiveness, and tumorigenesis were observed and compared in cells before and after ncRNAs enhancement. After microRNA-10b and HOTAIR were enhanced separately, several cancer stem cell (CSC)-like traits appeared in these LNSCs, including in vitro-enhanced proliferative capacity, expression of putative LCSC markers, progressive invasive ability, and even in vivo aggravation into and taking the place of normal liver tissue. Furthermore, strengthened expression of these ncRNAs partially degraded E-cadherin in LNSCs, which is one of the classic markers in epithelial-to-mesenchymal transition (EMT). HOTAIR or miR-10b enhanced in LNSCs may drive the LNSCs to a tendency toward malignant transformation. This study partially uncovers the mechanism by which miR-10b or HOTAIR promotes malignant transformation of LNSCs through down-regulating E-cadherin and inducing EMT.