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 (p0.05). 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.

Effect of 90 percentage portal branch ligation on liver regeneration and expression of metalloproteinase and tissue inhibitors of metalloproteinases in rats / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effect of 90% portal branch ligation on liver regeneration and expression of metalloproteinases and tissue inhibitors of metalloproteinases in rats.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Hepatocellular glycogen alleviates hepatic ischemia reperfusion injury and its relationship to ICAM-1 gene expression / 中华肝脏病杂志

<p><b>OBJECTIVE</b>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).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>Hepatocellular glycogen is important in protecting liver ischemic reperfusion injury. Also hepatocellular glycogen decreases the expression of ICAM-1 mRNA of HSCs.</p>

Ultrasound-guided percutaneous tube drainage combined with directly-viewed debridement with cholangioscopy: a mini-invasive strategy for peripancreatic necrotizing infection / 中华外科杂志

<p><b>OBJECTIVE</b>To establish "an integrative therapy" of drainage and debridement on peripancreatic necrotizing infection (PPNI) with minimally invasive technique, and to detect its clinical effects.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Perspective of pre-operational jaundice-reducing indication in carcinoma of head of pancreas / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the necessity, advantages and disadvantages of reducing the Icterus Index before operation in carcinoma of the head of pancreas.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Design of slow-paced radio-frequency thermo-therapeutic system for liver cancer / 医疗卫生装备

Thermo-therapeutic has been proved to be an effective approach for cancer therapy.This paper introduces a kind of new equipment suitable for liver oncology based on this theory,which uses the RF field as the thermal energy and adopts sequential-delivering technology.Its principle and advantages are introduced in detail.

The experimental study on the oval cells participating in hepatocarcinogenesis / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the genesis of the oval cells in hepatic tissue and relationship between the hepatic oval cells and the primary hepatocarcinoma.</p><p><b>METHODS</b>Sixty SD rats were divided into normal group (20 cases) and experimental group (40 cases). C-kit and PCNA were continuously detected by immunohistochemistry and the liver pathologic changes were observed by optical microscope in the different period.</p><p><b>RESULTS</b>The hepatical surface was lubricous and the histological morpha was normal in the normal group, and the positive cells of C-kit and PCNA were found on occasion. In the experimental group, the oval cells which express C-kit and PCNA were firstly discovered in the periportal regions in the second weeks, and these cells proliferate in turn and rangely in the bile duct epithelium. With the hepatic injury becoming more serious, the oval cells extended into the centrilobular regions from periportal regions. When HCC occurred, the oval cells were found in the cancer nodule and in the margin. In this period, the most of positive cells of C-kit still located in the periportal regions, while the PCNA cells were found in and out of the cancer nodule.</p><p><b>CONCLUSIONS</b>The oval cells originate from bile duct of the periportal regions. The oval cells play an important role in the process of hepatocarcinogenesis.</p>

The role of p38 MAPK pathway in ischemia-reperfusion injury of isolated liver / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the effect of p38 MAPK activity on isolated rabbit liver during the period of cold preservation and reperfusion.</p><p><b>METHODS</b>Based on the cold preservation-reperfusion model of isolated rabbit livers, according to the concentration of SB202190 in the preservation solution which was a specific p38 MAPK inhibitor, the isolated livers were divided into 4 groups, six in each A, B, C and D. Liver tissue samples and blood samples were harvested at different time points: before and end of cold preservation, reperfusion for 5, 10, 15, 30, 60, and 120 minutes. The activity levels of p38 MAPK were detected by both western blot and immunoprecipitation. The function markers of isolated livers were detected with automatic biochemistry analyzer, and the levels of total bile after reperfusion were measured.</p><p><b>RESULTS</b>In normal rabbit liver tissues, p38 MAPK had low activity. During the cold preservation period, the activity of p38 MAPK elevated slightly. But during the reperfusion period, the activity of p38 MAPK changed markedly which elevated rapidly at the early stage and reached its peak value at 10 minutes, then decreased gradually to the normal level (7.6 +/-0.9) at 120 minutes. SB202190 could inhibit the activity in a dose-dependent manner. The peak values of p38 MAPK activity in group B,C and D were 42.5 +/-2.4, 10.1+/-1.4, and 7.6 +/-0.6 respectively, while 78.6 +/-6.1 in group A. During the reperfusion period, the levels of serum ALT, AST and ALP were higher in group A than those in any other group, alike the p38 MAPK activity, especially at 15 and 30 minutes. On the other hand, the total bile secretion volume was (10.2 +/-2.9) ml/100 g, (13.9 +/-1.3) ml/100 g, (15.6 +/-1.2) ml/100 g, and (16.0 +/-1.3) ml/100 g in group A, B, C and D respectively.</p><p><b>CONCLUSIONS</b>During the cold preservation- reperfusion period, p38 MAPK specific inhibitor SB202190 can lower the p38 MAPK activity, and ameliorate the isolated liver injury. Activation of p38 pathway is one of the important mechanisms to cause ischemia-reperfusion injury of isolated liver.</p>

Apoptosis in hepatoma cells induced by antisense oligodeoxynucleotide against survivin / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To investigate the apoptosis induced by antisense oligodeoxynucleotide (ASODN) against survivin and the mechanisms after the hepatocellular carcinoma SMMC-7721 cells transfected with the ASODN.</p><p><b>METHODS</b>The ASODN was transfected into SMMC-7721 cells mediated by liposomal reagent. The changes of cell cycle and apoptotic rate were detected by flow cytometry. The changes of cell skeleton was observed through confocal microscope. The activity of p38MAPK and caspase-3 were detected by immuno-precipitation and kinase activity assess methods, respectively.</p><p><b>RESULTS</b>There were control, sense control, 400, 600, 800, and 1 000 ng/ml ASODN groups (I - VI). The apoptotic rats were 0.70%, 0.76%, 2.43%, 7.82%, 23.11%, and 31.35% in groups I - VI, respectively, which in the ASODN-transfected groups were higher than that in the control group (t<or=-9.6, P<0.01). The cell cycle in the ASODN-transfected groups stopped at G2/M phase. The well-arranged structure of microfilament was broken or destroyed, and the average fluorescent intensity of actin were 189.69+/-6.68, 184.23+/-8.76, 173.14+/-8.15, 99.48+/-6.57, 76.69+/-10.05 and 63.80+/-6.79 in groups I - VI, respectively), which in groups IV - VI decreased markedly, compared with that in the control group (t>or=20.9, P<0.01). The activity of p38MAPK increased significantly, when the ASODN was transfected at the concentration of 600 ng/ml or more, so did the caspase-3 activity (the p38MAPK and caspase-3 activity in groups I - VI were 7.03, 7.07, 13.47, 16.37, 43.97, 47.87 and 0.015+/-0.010, 0.014+/-0.002, 0.026+/-0.003, 0.042+/-0.001, 0.093+/-0.001, 0.100+/-0.001, respectively).</p><p><b>CONCLUSIONS</b>ASODN targeting at survivin mRNA can induce G2/M stop, activate p38MAPK and caspase-3. The activated caspase-3 destroys the cell skeleton microfilament system, resulting in apoptosis.</p>