Gastro-protecting effect of gefarnate on chronic erosive gastritis with dyspeptic symptoms / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of gastro-protecting agents on symptomatic chronic gastritis is unclear. This multicenter, open, randomized trial was designed to compare the comprehensive effects of gefarnate with sucralfate on erosive gastritis with dyspeptic symptoms.</p><p><b>METHODS</b>Totally 253 dyspepsia patients confirmed with erosive gastritis were enrolled from six centers in China. They randomly received either daily 300 mg gefarnate or 3 g sucralfate for six weeks. The primary endpoint was the effective rate of both treatments on endoscopic erosion at week six.</p><p><b>RESULTS</b>Gefarnate showed an effective rate of 72% and 67% on endoscopic score and dyspeptic symptom release, which is statistically higher than sucralfate (40.1% and 39.3%, P < 0.001, intension-to-treat). For histological improvement, gefarnate showed both effective in decreasing mucosal chronic inflammation (57.7% vs. 24.8%, P < 0.001, intension-to-treat) and active inflammation (36.4% vs. 23.1%, P < 0.05, intension-to-treat) than the control. A significant increase of prostaglandins and decrease of myeloperoxidase in mucosa were observed in gefarnate group. Severity of erosion is non-relevant to symptoms but Helicobacter pylori (H. pylori) status does affect the outcome of therapy.</p><p><b>CONCLUSIONS</b>Gefarnate demonstrates an effective outcome on the mucosal inflammation in patients with chronic erosive gastritis. Endoscopic and inflammation score should be the major indexes used in gastritis-related trials.</p>

Effect of ecoimmunonutrition supports on maintenance of integrity of intestinal mucosal barrier in severe acute pancreatitis in dogs / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>One of the major causes of death in severe acute pancreatitis (SAP) is severe infection owing to bacterial translocation. Some clinical studies suggested that ecoimmunonutrition (EIN) as a new strategy had better treatment effect on SAP patients. But the experiment studies on the precise mechanism of the effect of EIN were less reported. In this study, we mainly investigated the effects of EIN on bacterial translocation in SAP model of dogs.</p><p><b>METHODS</b>SAP was induced by retrograde infusion of 5% sodium taurocholate into the pancreatic duct in healthy hybrid dogs. The SAP dogs were supported with either parenteral nutrition (PN) or elemental enteral nutrition (EEN) or EIN. The levels of serum amylase, serum aminotransferase and plasma endotoxin were detected before and after pancreatitis induction. On the 7th day after nutrition supports, peritoneal fluid, mesenteric lymph nodes (MLN), liver, and pancreas were collected for bacterial culture with standard techniques to observe the incidence of bacterial translocation. Pathology changes of pancreas were analyzed by histopathologic grading and scoring of the severity of pancreas, and the degree of intestinal mucosal damage was assessed by measuring mucosal thickness, villus height, and crypt depth of ileum.</p><p><b>RESULTS</b>Compared with PN and EEN, EIN significantly decreased the levels of serum amylase, serum aminotransferase, plasma endotoxin, and the incidence of bacterial translocation. Furthermore, compared with the others, the histology scores of inflammation in pancreas and the ileum injury (ileum mocosa thickness, villus height, and crypt depth) were significantly alleviated by EIN (P < 0.05). Moreover, concerning liver function, the serum levels of alanine aminotransferase, aspartate aminotransferase and albumin were ameliorating significantly in the EIN group.</p><p><b>CONCLUSION</b>Our results suggested that EIN could maintain the integrity of intestinal mucosal barrier and reducing the incidence of bacterial translocation in SAP dogs. Early EIN was safe and more effective treatment for SAP dogs.</p>

Cloning of Human canstatin gene and expression of its recombinant protein / 中国医学科学院学报

<p><b>OBJECTIVE</b>To clone human canstatin gene and express its recombinant protein.</p><p><b>METHODS</b>The total RNA was extracted from human placenta. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and transformed into E.coli DH5alpha through electroporation. The gene was sequenced by the Sanger Dideoxy-mediated chain-termination method, and then the canstatin cDNA was cloned into the BamHI and HindIII sites of plasmid pET-22b (+) and transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b-Dgalactopyranoside (IPTG).</p><p><b>RESULTS</b>The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting products were cloned into pUCm-T vectors, and then were transformed into E.coli DHSa. After an over night culture, both blue and white colonies were found on the agar plate. Six white colonies were selected and cut by BamHI and HindIII. The plasmids DNA in one white colony showed one band near the location of primary plasmid after digested by BamHI and two bands near the locations of primary plasmid and objective gene fragment after digested by HindIII. The cloned gene in this white colony was sequenced and demonstrated to have the same sequence as that of canstatin gene in GenBank. Then canstatin cDNA was cut down from pUCm-T with BamHI and HindIII and ligated into the vector pET-22b (+). The resultant plasmid pET-22b (+)/canstatin was then transformed into E.coli BL21. White colonies were found on LB agar plate. Seven of them were selected and their plasmids were digested with both BamHI and HindIII. After electrophoresis, all selected colonies showed two specific bands, one was found near the location of primary plasmids, and the other near that of objective gene fragment. After IPTG induction, there was a new protein band about Mr 24 000 on SDS-PAGE. As estimated by densitometry, the percentage of the expressed product over total bacterial proteins was 18.2%, 18.8%, 23.0% and 23.4%, respectively, 1, 2, 3, and 4 hours after induction.</p><p><b>CONCLUSION</b>Human canstatin gene was successfully cloned and its recombinant proteins were expressed in this study.</p>

Role of extracellular signal-regulated kinase 1/2 in pathogenesis of stress-induced gastric ulcer / 第二军医大学学报

Objective:To investigate the role of extracellular signal-regulated kinase 1/2(ERK1/2)signal pathway in the pathogenesis of stress-induced gastric ulcer.Methods:Animal model of stress-induced gastric ulcer was established in rats with water-immersion restraint(WIR)stress.The mucosal activation of ERK1/2 was observed before and 5,15 and 30 min,and 1, 2 and 3.5 h after WIR stress.Some animals were also treated with an intravenous injection of PD98059(1 mg/kg),a specific ERK1/2 inhibitor,1 h prior to WIR stress.Expression of total ERK1/2 and caspase-3 were detected by Western blot analysis; ERK1/2 activity was measured by kinase activity assay using myelin basic protein as a specific substrate.DNA-binding activities of the transcription factors activator protein-1(AP-1)and nuclear factor-?B(NF-?B)were determined by electrophoretic mobility shift assays(EMSA).Mucosal TNF-?and IL-1?mRNA expression was analyzed by Northern blot analysis.The degrees of the gastric mucosal lesions were expressed as ulcer index(UI)and pathological evaluation.Apoptosis in the gastric mucosa was examined by an in situ TdT-mediated dUTP nick-end labeling(TUNEL)method.Results:Activated ERK1/2 was very weakly expressed in the gastric mucosa of normal rats.ERK1/2 was rapidly activated in the gastric mucosa of rats 15 min after WIR stress and the activity reached the maximal after 3.5 h.Pretreatment with PD98059 significantly inhibited ERK1/2 activation,decreased AP-1 and NF-?B activities and TNF-?and IL-1?mRNA expression,and obviously relieved gastric mucosal lesions,accompanied by caspase-3 activation and increased apoptosis.Conclusion:The present results indicate that ERK1/2 activation plays an important role in the development of stress-induced gastric ulcer.