Objective To investigate the distribution of neurons expressing Fos protein in central nervous system (CNS) following esophageal mucosal acid exposure,and to map the contribution of spe- cific brain areas in sensitizing responsivity and emphasize the coding change of CNS to the esophageal acid stimulation.Methods Thirty-six healthy Sprague-Dwley rats were randomly divided into five groups. Group A (n=6) was normal group of home cage control animals to which no stimulation was given. Group B (n=7) was saline group which received esophageal perfusion with normal saline solution (0.9% NaCl).Group C (n=8) was treated with esophageal mucosal acid exposure containing 0.1 mol/L HCl. Group D (n=7) was sensitized by ovalbumin.Group E (n=8) received basal ovalbumin-sensitization plus esophageal mucosal acid exposure.The rat model of esophageal visceral hypersensitivity was estab- lished by the basic ovalbumin-sensitization combined with intra-esophageal mucosal acid exposure.The neuronal expressions of c-fos proto-oncogene were detected with immunohistochemical counter-staining and computerized color image analyzer under various conditions.Results The rats in model group with basic ovalbumin-sensitization plus esophageal acid perfusion initiated a high density expression of c-fos- immunoreactive(Fos-IR) neurons in multineuronal networks.A significantly higher number of Fos positive neurons was found in the model group than those in the corresponding regions of other groups (P＜0.05) in the following brain areas:frontal and parietal cortex,insular cortex,cingulated cortex,central amyg- daloid nucleus,the K(?)lliker-Fuse nucleus,the nucleus ambiguus,parabrachial nucleus,hypothalamic paraventricular nucleus,paraventricular thalamic nucleus,paratrigeminal nucleus,the nucleus of solitary tract,area postrema,reticular nucleus of medulla,whereas no significant difference was found in the dorsal motor nucleus of the vagus,supraoptic nucleus,periaqueductal gray matter or orbital part of infe- rior frontal gyrus.The values of Fos-IR neurons were also increased in the central amygdala,parabrachi- al nucleus,paraventricular nucleus,the paratrigeminal nucleus and NTS in the model group than that in the corresponding regions of other groups (P＜0.05).Conclusion The basic ovalbumin-sensitization fa- cilitated dramatically the c-fos expression evoked by esophageal acid perfusion,suggesting that visceral hypersensitivity induced by ovalbumin may alter cortical reactivity processing of esophageal acid stimula- tion in brain areas.

Humans ; Telomere ; metabolism ; Telomere-Binding Proteins ; metabolism

Carcinoma, Hepatocellular ; genetics ; DNA, Mitochondrial ; genetics ; Humans ; Liver Neoplasms ; genetics ; Microsatellite Repeats

Objective To assess mucin gene expression in Barrett's esophagus.Methods Mucin core protein-MUC1,MUC2,MUC3,MUCSAC and MUC6 were detected by immunohistochemistry.The re- lationship between mucin expression and magnification-endoscopic characteristics,pathohistologic epithelial types of Barrett's esophagus was analyzed.Results Mild expression of MUC1 was predominantly found in the superficial epithelium of both gastric and specialised intestinal metaplasia.In a small number of specimens, mild expression of MUC1 was also noted in glands.Strong MUC2 expression was noted only in the goblet cells in Barrett's oesophagus.MUC3 was expressed in the superficial columnar cells of specialized intestinal metaplasia with or without globlet cells but not in gastric metaplasia of the oesophagus.In some specimens MUC3 was expressed in the vacuolus of the globlet cells and the lumen of gland.Strong staining of MUCSAC was noted in the columnar epithelium of both gastric metaplasia and specialized intestinal metaplasia in Barrett's oesophagus,as well as expressed in the cytoplasm and vacuolus of the globlet cells in some speci- mens.Expression of MUC6 protein was detected at the basement of the crypts in gastric metaplasia and spe- cialised Barrett's glands.Expression of MUC2 and MUC3 protein was found much higher in villous or irregu- lar pit pattern than that in dot or rod pit pattern(P

Objective To investigate characteristics and alternation of cerebral evoked potentials (CEP) response to esophageal mucosal acid exposure and distention in patients with non-erosive gastro-oesoph- ageal reflux disease (NERD) and in healthy subjects,and to study the mechanism of visceral hypersensitivity in NERD.Methods Twenty-one NERD patients and 10 volunteers were recruited.Mechanical distention stimulation and acid perfusion of the esophagus were performed using the balloon-affixed and polyvinyl multi- lumen catheter.First,maximally tolerated pain thresholds of all subjects were recorded,then esophageal mechanical stimulation with a 75% of maximal tolerated intensity and a frequency of 0.2 Hz was performed altogether 64 times by means of a computer-controlled barostat.The alternation of esophageal CEP was recorded before and after acid perfusion with a multichannel international 10-20 system of electroencephalography. Experimental data was analyzed by student's t-test and one way analysis of variance.Results Esophageal mu- cosal distention may evoke recognizable and reproducible and multi-peak CEP.The latencies for N1,P1 and N2 in volunteers were (246?77),(388?84)and (502?78) ms,CEP morphology of NERD patients was charac- terized by randomly distributed patterns,and the latencies for N1 ,P1 and N2 were (192?46),(293?76) and (440?79)ms,significantly shorter for mechanical stimulation compared with those of control group respectively (all P value

OBJECTIVESTo construct a novel virus-like particulate peptide-nucleic acid vaccine (VPNV) of human telomerase reverse transcriptase (hTERT), and to study its anti-liver cancer immunity.

METHODSA cationic antigenic peptide was synthesized and purified, and then human granulocyte macrophage colony stimulating factor (hGM-CSF) and TERT gene were cloned into the eukaryotic expression vector pTCAE. The peptide was combined with the nucleic acid vaccine to make a VPNV, which was transfected into eukaryotic cell COS-7. The immunogenicity of hGM-CSF and hTERT were detected using ELISA and Western blot. The efficacy of VPNV for inducing antigen specific CTL response was determined using the lactate dehydrogenase release method.

RESULTSVPNV was verified capable to trigger specific CTL responses and has shown a specific cytolytic activity to liver cancer cell HepG2.

CONCLUSIONA VPNV which can stimulate antigen specific CTL response was successfully constructed. This paves the way for our further investigation of anti-liver cancer immunity in mice.

Animals ; Cancer Vaccines ; immunology ; Cloning, Molecular ; Eukaryotic Cells ; metabolism ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; immunology ; Liver Neoplasms ; immunology ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Peptide Nucleic Acids ; genetics ; immunology ; Telomerase ; genetics ; immunology ; Vaccines, DNA ; immunology

OBJECTIVETo review the recent research progress in pharmacological actions and mechanisms of ginkgolide B. Data sources Information included in this article was identified by searching of PUBMED (1987 - 2006) online resources using the key terms "ginkgolide B", "platelet activating factor", and "pharmacological". Study selection Mainly original milestone articles and critical reviews written by major pioneer investigators of the field were selected.

RESULTSThe key issues related to the pharmacological actions and mechanisms of ginkgolide B were summarized. The ginkgolide B possesses a number of beneficial effects such as anti-inflammatory, anti-allergic, antioxidant, and neuroprotective effects. Meantime, their mechaniams were discussed.

CONCLUSIONSThe Ginkgolide B is the most potent antagonist of platelet activating factor (PAF) and exhibits therapeutic action in a variety of diseases mainly by the PAF receptor.

Acute Disease ; Animals ; Anti-Asthmatic Agents ; pharmacology ; Antineoplastic Agents ; pharmacology ; Ginkgolides ; chemistry ; pharmacology ; Humans ; Lactones ; chemistry ; pharmacology ; Neuroprotective Agents ; pharmacology ; Pancreatitis ; drug therapy ; Reperfusion Injury ; prevention & control

Esophageal and Gastric Varices ; complications ; Gastrointestinal Hemorrhage ; etiology ; surgery ; Hepatic Encephalopathy ; epidemiology ; etiology ; prevention & control ; Hepatorenal Syndrome ; etiology ; surgery ; Humans ; Hypertension, Portal ; complications ; surgery ; Portasystemic Shunt, Transjugular Intrahepatic ; adverse effects ; methods ; Postoperative Complications ; Treatment Outcome

OBJECTIVESTo investigate the mechanisms for human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) in increasing hepatocellular carcinoma cell apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL).

METHODSCell apoptosis was identified by flow cytometry analysis after annexin V/PI double staining. Expression of apoptosis-related proteins, procaspase-8, -9, -3, Bax, Bcl-2 and hTERT, were identified by Western blotting analysis; telomerase activity and telomere length were detected by telomeric repeat amplification protocol (TRAP) and telomere amount and length assay (TALA) methods.

RESULTSHepatocellular carcinoma cell apoptosis induced by TRAIL were all significantly increased by hTERT RNAi (P less than 0.05). For example, apoptosis rates were enhanced from 5.53% (untransformed) to 10.35% (transformed) in HepG 2 cells and from 14.73% to 77.24% in SMMC 7721 cells after being treated by 100 ng/ml TRAIL for 24 h. Moreover, activation of procaspase-8, -9 and -3 in transformed cells after being treated by TRAIL were all significantly raised (P less than 0.05) in a dose-dependent manner. The expression of procaspase-8, -9 and Bcl-2 were effectively augmented (P less than 0.05), but expressions of Bax and hTERT were strikingly decreased (P less than 0.05). Meanwhile, telomerase activity was apparently suppressed and telomere length was markedly shortened (P less than 0.05). There were no remarkable differences in these effects between control cells and the untransformed cells (P more than 0.05).

CONCLUSIONEnhanced cell apoptosis induced by TRAIL through hTERT RNAi may be related to up-regulation of procaspase-8 and -9 expressions. However the down-regulation of hTERT expression, reduced telomerase activity and shortened telomere length may not be related to expressions of Bcl-2 and Bax.

Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Telomerase ; genetics ; metabolism

BACKGROUND/AIMS: To investigate the differential expression of RING finger (RNF) proteins in Barrett esophagus (BE) and esophageal adenocarcinoma (EAC). METHODS: The differential expression of RNFs in normal esophagus (NE), BE, and EAC was screened using microarray assay. Real-time quantitative polymerase chain reaction (PCR), tissue microarray assay, and Western blot analysis were independently performed to detect the mRNA and protein expression of screened RNFs. RESULTS: The expression of nine RNFs in the BE or EAC was 2-fold higher than those in NE. Among these proteins, the RNF32 and RNF121 expression in BE was 20.3-fold and 16.4-fold higher, respectively, than that in NE, and the expression of RNF24, RNF130, RNF141, RNF139, RNF11, RNF14, and RNF159 was upregulated more than 2-fold compared with NE. The expression of nine RNFs was not only upregulated in the EAC but was also positively related to the RNF expression in BE. The PCR results also indicated increased expression of these RNFs in BE and EAC compared to NE. Furthermore, the mRNA expression of all RNFs, except for RNF141 in EAC, was dramatically higher than those in the BE. Similar results were also obtained from the Western blot analysis. CONCLUSIONS: A total of nine RNFs play critical roles in the progression of BE to EAC.
Adenocarcinoma/*enzymology/genetics ; Adult ; Aged ; Barrett Esophagus/*enzymology/genetics ; Carrier Proteins/genetics ; Disease Progression ; Esophageal Neoplasms/*enzymology/genetics ; Female ; Gene Expression Profiling ; Humans ; Intracellular Signaling Peptides and Proteins/genetics ; Male ; Middle Aged ; Proteins/genetics ; *RING Finger Domains ; Receptors, Cell Surface/genetics ; Ubiquitin-Protein Ligases/genetics/*metabolism