Abdominal puncture drainage alleviates severe acute pancreatitis in rats by activating Nrf-2/HO-1 pathway and promoting autophagy / 南方医科大学学报

OBJECTIVE@#To assess the effect of early abdominal puncture drainage (APD) on autophagy and Nrf-2/HO-1 pathway in rats with severe acute pancreatitis (SAP) and explore the possibile mechanism.@*METHODS@#Thirty-two male SD rats were randomly divided into sham-operated (SO) group, SAP group with retrograde injection of 4% sodium taurocholate, APD group with insertion of a drainage tube into the lower right abdomen after SAP induction, and APD + ZnPP group with intraperitoneal injection of 30 mg/kg ZnPP 12 h before APD modeling. Blood samples were collected from the rats 12 h after modeling for analysis of amylase and lipase levels and serum inflammatory factors. The pathological changes of the pancreatic tissue were observed with HE staining. Oxidative stress in the pancreatic tissue was detected with colorimetry, and sub-organelle structure and autophagy in pancreatic acinar cells were observed by transmission electron microscopy. The expressions of autophagy-related proteins and Nrf-2/HO-1 pathway were detected using RT-PCR and Western blotting.@*RESULTS@#Compared with those in SAP group, the rats with APD treatment showed significantly alleviated pathologies in the pancreas, reduced serum levels of lipase, amylase and inflammatory factors, lowered levels of oxidative stress, and activated expressions of Nrf-2/HO-1 pathway in the pancreas. The ameliorating effect of ADP was significantly inhibited by ZnPP treatment before modeling. APD obviously reversed mitochondrial and endoplasmic reticulum damages and p62 accumulation induced by SAP.@*CONCLUSION@#APD treatment can suppress oxidative stress and repair impaired autophagy in rats with SAP by activating the Nrf-2/HO-1 pathway, thereby reducing the severity of SAP.

Role of CaMK II in pancreatic injury in mice with severe acute pancreatitis / 南方医科大学学报

OBJECTIVE@#To investigate the expression of Ca2+/calmodulin-dependent protein kinase II (CaMK Ⅱ) in pancreatic tissues of mice with severe acute pancreatitis (SAP) and explore the protective effect of KN93, a CaMK Ⅱ inhibitor, against pancreatic injury in SAP and the possible mechanism.@*METHODS@#Thirty-six healthy male C57 mice were randomly divided into sham operation group, SAP group, KN93 group and SAP + KN93 group (n=9). Serum and pancreatic tissue samples were collected 24 h after modeling. The pathological changes in the pancreatic tissues were observed using HE staining. Serum lipase and amylase activities and the levels of inflammatory factors were detected using ELISA. Western blotting was used to detect the expressions of CaMK Ⅱ, p-CaMK Ⅱ, p-NF-κB, MAPK and p-MAPK in mouse pancreas.@*RESULTS@#Compared with those in sham operation group, the expressions of p-CaMK Ⅱ, p-NF-κB and p-MAPK were significantly increased in SAP group (P < 0.05). KN93 treatment obviously alleviated pathological injuries of the pancreas in SAP mice, and significantly lowered serum levels of lipase, amylase and inflammatory factors (TNF-α and IL-6) and phosphorylation levels of NF-κB, ERK and MAPK proteins (P < 0.05).@*CONCLUSION@#The activity of CaMK Ⅱ is significantly increased in the pancreatic tissue of SAP mice. KN93 can alleviate pancreatic injury and inflammation in SAP mice possibly through the ERK/MAPK signaling pathway.

Research progress of corneal thickness measurement / 国际眼科杂志(Guoji Yanke Zazhi)

@#The thickness of the central cornea has an important influence on various eye diseases and operations such as keratoconus and other corneal diseases, glaucoma, and corneal refractive surgery. Obtaining accurate central corneal thickness is a topic that clinicians have been paying close attention to. It decides the operation method and operation parameters(cutting depth, cutting optical area size, <i>etc.</i>)of refractive surgery. Accurate measurement of central corneal thickness is a great concern to clinicians. At present, there are two kinds of measurement of corneal thickness: the first is ultrasonic measurement, such as traditional Type A ultrasonic corneal thickness meter and ultrasonic biological microscope; the second is optical measurement, including Pentacam, corneal endokeratoscope, optical coherence tomography, <i>etc</i>. Different measuring methods and instruments have their own advantages and disadvantages. However, the ultimate goal of developing corneal thickness measurement is easy operation and good repeatability. Therefore, based on the summarization of current clinically-used corneal thickness measurement instruments and of research progress of corneal thickness measurement, this paper aims at providing theoretical guide for clinical oculists.

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.

Effects of Yinzhihuang oral liquid and Lonicera japonica extract on hemolysis and hyperbilirubinemia in rats with glucose-6-phosphate dehydrogenase deficiency / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effects of different doses of Yinzhihuang oral liquid and different concentrations of Lonicera japonica extract on hemolysis and hyperbilirubinemia in rats with glucose-6-phosphate dehydrogenase (G6PD) deficiency.</p><p><b>METHODS</b>Male Wistar rats were randomly divided into 10 groups (n=10 each): normal control group (untreated), negative control group (saline-treated), positive control group (primaquine-treated), low-, medium- and high-dose Yinzhihuang oral liquid groups (13.4, 26.8, and 53.6 mL/kg, respectively), and low-, medium-, high-, and very-high-concentration Lonicera japonica groups (6.7 mL/kg administered, containing 8, 40, 80, and 160 mg/mL Lonicera japonica extract, respectively). A rat model of acetylphenylhydrazine-induced G6PD deficiency was established in all groups except the normal control group, as confirmed by the morphological changes in erythrocytes observed using Wright's stain. After treatment, routine blood and biochemical tests were conducted to measure hemolytic indices, as well as changes in total and indirect bilirubin levels.</p><p><b>RESULTS</b>Rats with G6PD deficiency demonstrated irregular erythrocytes with a lighter-staining center. In the positive control group, the red blood cell count decreased, while the free hemoglobin count and the reticulocyte percentage increased, as compared with before treatment (P<0.05); in all the Yinzhihuang oral liquid groups and Lonicera japonica extract groups, all the above indices except reticulocyte percentage returned to the levels before treatment (P<0.05). Compared with the positive control group, all the Yinzhihuang oral liquid groups had significantly reduced total and indirect bilirubin levels (P<0.05), and all the Lonicera japonica group had significantly reduced indirect bilirubin levels (P<0.05). However, the total bilirubin level was significantly higher in the Lonicera japonica groups than in the Yinzhihuang oral liquid groups (P<0.05). The low-dose Yinzhihuang oral liquid group demonstrated a significantly greater decrease in total bilirubin level than the medium- and high-dose Yinzhihuang oral liquid group (P<0.05).</p><p><b>CONCLUSIONS</b>Administration of high-dose Yinzhihuang oral liquid and different concentrations of Lonicera Japonica extract do not cause hemolysis in rats with G6PD deficiency. Yinzhihuang oral liquid is more effective in treating hyperbilirubinemia than Lonicera Japonica extract. However, the efficacy of Yinzhihuang oral liquid may not be dose-dependent.</p>

Effect of rhubarb on neonatal rats with bronchopulmonary dysplasia induced by hyperoxia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effect of rhubarb on neonatal rats with bronchopulmonary dysplasia (BPD) induced by hyperoxia.</p><p><b>METHODS</b>A total of 64 rats (postnatal day 4) were randomly divided into four groups: air control, rhubarb control, hyperoxia model, and hyperoxia+rhubarb (n=16 each). The rats in the hyperoxia model and hyperoxia+rhubarb groups were exposed to hyperoxia (60% O2) to establish a BPD model. The rats in the rhubarb control and hyperoxia+rhubarb groups were given rhubarb extract suspension (600 mg/kg) by gavage daily. The pathological changes of lung tissue were evaluated by hematoxylin-eosin staining on postnatal days 14 and 21. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were measured by spectrophotometry. The mRNA and protein expression levels of tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were determined by RT-PCR and Western blot respectively.</p><p><b>RESULTS</b>The hyperoxia model group showed reduced alveolar number, increased alveolar volume, and simplified alveolar structure, which worsened over the time of exposure to hyperoxia. These pathological changes were significantly reduced in the hyperoxia+rhubarb group. On postnatal days 14 and 21, compared with the air control and rhubarb control groups, the hyperoxia model group had significantly reduced radical alveolar count (RAC), significantly reduced activity of SOD in the lung tissue, and significantly increased content of MDA and mRNA and protein expression levels of TNF-α and IL-6 (P<0.05). Compared with the hyperoxia model group, the hyperoxia+rhubarb group had significantly increased RAC, significantly increased activity of SOD in the lung tissue, and significantly reduced content of MDA and mRNA and protein expression levels of TNF-α and IL-6 (P<0.05).</p><p><b>CONCLUSIONS</b>Rhubarb may play a protective role in rats with BPD induced by hyperoxia through inhibiting inflammatory response and oxidative stress.</p>

The immune response in acute and chronic pancreatitis / 解放军医学杂志

Acute and chronic pancreatitis are most common gastrointestinal diseases.Recently,there are emerging evidences that immune cell play important roles in the pathogenesis of acute and chronic pancreatitis.Studies have shown that macrophages,high mobility group protein 1 (HMGB 1) and interleukin-33 (IL-33) play an important role in the pathological process of pancreatitis,and are regulated by multiple levels.For example,immune cells are critical in the development and progression of pancreatitis,which not only have the ability to induce microenvironment,but also respond to danger signals derived from endogenous and exogenous molecules.Therefore,further understanding of relevant immune signaling will provide new idea and potential therapeutic targets that can prevent disease progression.Here,we review recent data from animal and human clinical studies that focus on immune responses.

A comparative analysis of neonatal critical illness score and score for neonatal acute physiology, perinatal extension, version II / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the accuracy and clinical utility of neonatal critical illness score (NCIS) and score for neonatal acute physiology, perinatal extension, version II (SNAPPE-II) in predicting the "dead and abandoned" risk in critically ill neonates.</p><p><b>METHODS</b>A total of 269 critically ill neonates were divided into two groups according to their prognosis: dead/abandoned and improved/cured. The accuracy of these two scoring systems, NCIS and SNAPPE-II, in predicting the "dead and abandoned" risk was compared.</p><p><b>RESULTS</b>The dead/abandoned group had a significantly higher SNAPPE-II score than the improved/cured group (P<0.001), while there was no significant difference in the NCIS score between the two groups (P=0.091). The children who were in line with the individual indicator in the NCIS results had a significantly higher "dead and abandoned" risk than those who were not (P=0.005).</p><p><b>CONCLUSIONS</b>SNAPPE-II is more accurate in early prediction of the "dead and abandoned" risk in critically ill neonates compared with NCIS. NCIS has the ability to predict the "dead and abandoned" risk in children in line with the individual indicator.</p>

Influence of gastric bypass surgery on hepatic gluconeogenesis in type 2 diabetic Goto Kakizaki rats / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the influence and significance of gastric bypass surgery on hepatic gluconeogenesis in type 2 diabetic Goto Kakizaki(GK) rats.</p><p><b>METHODS</b>Forty GK rats were randomly divided into Roux-en-Y gastric bypass group(group A) and sham operation group(group B). Differences in glucose tolerance experiment(OGTT) at preoperative and postoperative 1, 2 and 4 weeks were compared and weight was recorded. Glycated hemoglobin levels were measured preoperatively and 4 weeks postoperatively. The animals were sacrificed 4 weeks after surgery and liver tissues were harvested to detect the relative expression of mRNA and protein of glucose 6 phosphatase(G-6-P) and phosphoenol pyruvate kinase(PEPCK) with RT-PCR and Western blot.</p><p><b>RESULTS</b>Fasting blood glucose levels were 6.5, 4.9, and 4.7 mmol/L in group A, and were 10.3, 10.4, and 12.5 mmol/L in group B, and the differences between two groups were statistically significant(P<0.05). The blood glucose level at 2 h after stomach lavage were 8.3, 6.4 and 5.5 mmol/L in group A, and were 21.4, 23.8 and 24.7 mmol/L in group B at postoperative 1, 2, 4 weeks, and the differences between two groups were statistically significant(P<0.05). The glycosylated hemoglobin at postoperative 4 weeks was(6.8±1.0)%, significantly lower than that in group B[(7.9±0.8)%, P<0.05]. Hepatic G-6-P and PEPCK mRNA relative expression at postoperative 4 weeks was reduced by 21.0% and 25.9% respectively as compared to group B, and the protein expression reduced as well. Immunohistochemistry showed that hepatic glycogen sedimentary in group A increased significantly.</p><p><b>CONCLUSION</b>The relative mRNA and protein level of key enzymes of hepatic gluconeogenesis are significantly decreased after Roux-en-Y gastric bypass surgery and hepatic gluconeogenesis is reduced, which may be a potential mechanism of the decrease of blood glucose.</p>

Levels of cytokines in peripheral blood of premature infants with chronic lung disease / 中国当代儿科杂志

<p><b>OBJECTIVE</b>This study examined the levels of interleukin (IL)-1β, IL-6, IL-8 and tumor necrosis factor alpha (TNF-α) in peripheral blood of premature infants with chronic lung disease (CLD) in order to investigate the possible role of immunologic factors in CLD.</p><p><b>METHODS</b>Twenty-six premature infants who had been admitted to the neonatal intensive care unit for 28 days were classified into CLD (n=14) and non-CLD (control, n=12) groups. IL-1β, IL-6, IL-8 and TNF-α concentrations in peripheral blood were measured by multiplex technique (Bio-plex).</p><p><b>RESULTS</b>There were no significant differences in peripheral blood levels of IL-1β, IL-6, IL-8 and TNF-α between the two groups.</p><p><b>CONCLUSIONS</b>There are no significant changes in peripheral blood levels of IL-1β, IL-6, IL-8 and TNF-α in premature infants with CLD on the non-acute phase. Immunologic factors might not play a key role in CLD on the phase.</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>

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>

Molecular features and expression of DAZAP2 in human multiple myeloma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>In our previous study, we found that DAZAP2 was the most significantly down regulated gene when differential screening of complementary DNA (cDNA) chips were used to analyze mRNA isolated from bone marrow mononuclear cells from newly diagnosed multiple myeloma (MM) patients without anticancer treatment. In this study, we observed DAZAP2 mRNA and protein expression in the mononuclear cells from MM bone marrow and investigated its role in the pathogenesis of MM.</p><p><b>METHODS</b>The full-length cDNA of DAZAP2 was cloned and sequenced from mononuclear cells from human bone marrow. The nucleotide and amino acid sequences of DAZAP2 were analyzed using the ClustalW program. A dendrogram was constructed by multiple sequence alignment using ClustalW and amino acid sequence identity/similarity was derived based on comparisons attained using the MegAlign software. The recombinant pEGFP expression vector was constructed and the confocal microscopy was used for the localization of the DAZAP2 protein in transfected COS7 cells. The expression of DAZAP2 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and the expression level of DAZAP2 protein was detected by Western blotting analysis in MM samples.</p><p><b>RESULTS</b>DAZAP2 proteins of vertebrates is highly conserved in evolution. It contains a proline-rich region, several potential SH2 and SH3 domain-binding motifs and a possible protein kinase C (PKC) phosphorylation site. We showed by confocal microscopy that the DAZAP2 protein predominantly resides in the cytoplasm with a discrete pattern of punctuated distribution. The expression of DAZAP2 was not detected in 24 of 36 MM samples by semi-quantitative RT-PCR. In contrast, DAZAP2 expression was detected in all 30 normal controls. The expression level of DAZAP2 protein was assayed by Western blotting analysis, showing a robust down-regulation in MM patients (P < 0.001) that matched with the results of the RT-PCR.</p><p><b>CONCLUSIONS</b>DAZAP2 is downregulated in MM samples and it may be a signal molecule in MM cells. DAZAP2 is involved in the pathogenesis of MM and could be used as a genetic marker for MM.</p>

Detection of MSH2 gene mutation by PCR / 中南大学学报(医学版)

OBJECTIVE@#To establish a genetic diagnosis method for a novel MSH2 mutation.@*METHODS@#A specific primer on the mutated site of MSH2 was synthesized and PCR was conducted using the specific primer and another downstream primer. PCR products were electrophoresed and then the carriers with the novel gene mutation of the carriers or non-carriers were identified.@*RESULTS@#MSH2 in a hereditary nonpolyposis colorectal cancer family were successfully found.@*CONCLUSION@#The method is effective and simple for genetic diagnosis of the novel mutation in MSH2.

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>

Human bone marrow multipotent adult progenitor cells differentiate into hepatocyte-like cells with hepatocyte growth factor plus fibroblast growth factor-4 in vitro / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the possibility of the human bone marrow multipotent adult progenitor cells (hMAPCs) to differentiate into hepatocytes with hepatocyte growth factor (HGF)/ fibroblast growth factor-4 (FGF-4) in vitro.</p><p><b>METHODS</b>(1) Obtaining the hMAPCs. Bone marrow was obtained from volunteers and then centrifuged through density gradient centrifugation methods. The collected mononuclear cells were cultured through adheret culture to get mesenchymal stem cells (MSCs). The hMAPCs were obtained through collecting and isolating the MSCs by magnetic activated cell sorting (MACS) through depletion selection by use of CD45 and GlyA microbeads. (2) Differentiation of the hMAPCs with HGF+FGF-4. Group A: HGF (20 ng/ml) + FGF-4 (10 ng/ml) induced hMAPCs; group B (positive control group): L-02 human hepatocytes(cell lines); and group C (negative control group): the undifferentiated hMAPCs. (3) The expressions of albumin (Alb), alpha fetoprotein (AFP), cytokeratin-18 (CK-18), and cytokeratin-19 (CK-19) were detected with immunocytochemistry to identify the characteristics of the differentiated cells at different times and the ratio of the positive cells was determined. (4) ALB, AFP, CK-18, and CK-19 expressions of the differentiated cells were detected by RT-PCR assay to investigate the mRNA transcriptions of characteristic hepatic proteins. (5) Alb expressions of the differentiated cells at different times were detected by Western blot on the 21st and 35th days.</p><p><b>RESULTS</b>(1) The results of immunocytochemistry. The staining of Alb, CK18 were essentially positive in group A. As an early marker of immature hepatocytes, AFP staining was positive on the 7th day but negative in later differentiating periods in group A. (2) The results of RT-PCR. On the 7th day, the differentiated hMAPCs expressed AFP mRNA but were negative in later differentiating periods. On the contrary, the mRNA of Alb and CK-18 were positive at all times. (3) The results of Western blot assay. Alb protein was positive on the 21st day and 35th day.</p><p><b>CONCLUSIONS</b>Under some definite inducing conditions hMAPCs can differentiate into hepatocyte-like cells. They may serve as a potential cell source for liver engineering.</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>