OBJECTIVETo explore a method which can remove the gastric mucus in order to prepare mucous membrane single cell suspension for the research of cytomics.

METHODSEnzymology was used to remove the mucus gel and to separate mucous layer from the normal fresh gastric tissue. The mucous layer was broken to prepare single cell suspension with machine method. Expression of major cyclins in mucous layer cells was examined by cytoimmunochemistry, flow cytometry(FCM) and confocal microscopy.

RESULTSThe 0.1% pepsin could dissolve the mucus gel and 1.2-2.4 U/L dispase could separate the mucous layer completely. The single mucous cell suspension was prepared successfully. FCM results from mucous single cell suspension revealed that expression of cyclin D(3), B(1) was obvious, that of cyclin D(2) was weak and that of cyclin D(1), A, E was the least. Similar results were found with confocal microscopy.

CONCLUSIONSSingle cell suspension from mucous layer can be easily prepared by pepsin and dispase. Cyclins schedule expression in vivo is different from cyclins schedule expression in vitro.

Cell Line ; Cell Proliferation ; Cyclins ; metabolism ; Flow Cytometry ; Gastric Mucins ; metabolism ; Gastric Mucosa ; cytology ; metabolism ; Humans ; Mucous Membrane ; cytology ; metabolism

OBJECTIVETo clone core promoter regions of iroquois homeobox gene 1 (IRX1) gene and evaluate the regulatory mechanism of IRX1 transcription.

METHODSUpstream sequence from transcriptional start site was predicted using bioinformatics methods. Serial deleted fragments from IRX1 promoter sequences were amplified by PCR and luciferase reporter plasmids were constructed. The luciferase intensity was analyzed after transferring reporters into GES-1 gastric mucosa cell line.

RESULTSThe promoter of IRX1 was predicted to be within 700 bp from the 5'-flanking region of IRX1 gene. Eight serial deleted luciferase reporter plasmids were constructed. The transcriptional activity of different fragments was expressed as following: p-416>p-584>p-715>p-350>p-687>p-320>p-188>p-92. Except p-320 and p-188, the transcriptional activity of other 6 fragments was higher than that of PGL3-basic plasmid. The transcriptional activity was the highest in p-416 and decreased sharply from p-320 to p-188.

CONCLUSIONSThe fragment p-416 shows the strongest promoter activity. The sequence from -320 bp to -188 bp is identified as core promoter region, which is focused as key sequence for further regulatory analysis, since some binding sites for important transcriptional factors such as Sp1 and TFII D are predicted.

Cell Line ; Cloning, Molecular ; Gastric Mucosa ; cytology ; Genes, Homeobox ; Homeodomain Proteins ; genetics ; Humans ; Promoter Regions, Genetic ; Transcription Factors ; genetics

Acetylcholine ; pharmacology ; Adenocarcinoma ; metabolism ; Cell Line, Tumor ; Gastric Mucosa ; cytology ; drug effects ; metabolism ; Humans ; Receptors, Cholinergic ; metabolism ; Stomach Neoplasms ; metabolism

AIMTo investigate the correlation between the gastric adaptive cytoprotection and the low concentration alcohol intake in a chronic drinking rat model and the effect of chronic ethanol exposures on the cell turnover of the gastric mucosa and its possible role in adaptive cytoprotection.

METHODSSprague-Dawley rats received the drinking water containing 6% (v/v) ethanol as their only water intake for 28 days. In the different stages of the 28 days (1st, 3rd, 7th, 14th and 28th days), the stomachs of the rats were cannulated and perfused with pure ethanol, and the severity of mucosal lesions was measured in 2 hours at the end of perfusion respectively. The cell proliferation and apoptosis in gastric mucosa of rats in different groups were analyzed by flow cytometer, immunohistochemistry and computer image analysis.

RESULTSPure ethanol caused ulcer and haemorrhagic damage in the corpus and antral mucosa of the control rats. These lesions were prevented by pretreatment of the animals with ethanol exposure in the 3 rd to 14 th days. The damage index was decreased by 80%, as compared with those in control rats. There was no significant difference in the rats exposed to the ethanol in the 1st and 28th days. Compared with control, the cell apoptosis in gastric mucosa of the rats was enhanced during they exposure to the ethanol in the 3rd to 28th days. Otherwise the cell proliferation was increased in the 3rd to 28th days, and decreased in the 28th days, respectively.

CONCLUSIONChronic adequate alcohol intake may enhance the cell turnover of gastric mucosa and lead to an adaptive cytoprotection. Long-term stimulus with the low concentration ethanol may cause the atrophy of gastric mucosa and reduce the gastric mucosal cytoprotective effect.

Alcoholism ; Animals ; Apoptosis ; Cell Proliferation ; Cytoprotection ; Ethanol ; adverse effects ; Gastric Mucosa ; cytology ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

The effect of acupuncture in the treatment of young pigs with induced enteropathogenic Escherichia coli diarrhea was histopathologically evaluated by routine hematoxylin and eosin stain. Thirty two pigs weighed 4-5kg and aged 21days old were used in this study. The animals with diarrhea were treated with traditional acupuncture, or enrofloxacin. In the group treated with traditional acupuncture, acupoint GV1 (Jiaochao) was used and in the group treated with antibiotics, enrofloxacin was injected intramuscularly. Ten pigs were inoculated with E. coli, but were not treated and served as nontreated control group. At postinoculation day 6, all pigs of the acupuncture and antibiotic treated groups recovered from diarrhea. In the ascending and descending colons of the nontreated control group, severe infiltration of inflammatory cells in the lamina propria was observed and in the fundic stomach, destruction of the fundic gland architecture and necrotic lesions were observed, however, in the same sites of the acupuncture and antibiotics treated groups, the mucosae of the colon and stomach were relatively similar to those of the normal group. These results indicate that acupuncture treatment is effective in controlling induced E. coli diarrhea in pigs at its early stage.
Acupuncture ; Animals ; Colon/cytology/microbiology/pathology ; Diarrhea/therapy/*veterinary ; Escherichia coli Infections/therapy/*veterinary ; Gastric Mucosa/cytology/microbiology/pathology ; Intestinal Mucosa/cytology/microbiology/pathology ; Male ; Stomach/cytology/microbiology/pathology ; Swine ; Swine Diseases/*microbiology/therapy

Acetylcholine ; pharmacology ; Cell Line, Tumor ; Cell Membrane ; Epithelial Cells ; cytology ; Gastric Mucosa ; cytology ; drug effects ; metabolism ; Humans ; Receptors, Nicotinic ; drug effects ; metabolism ; Stomach Neoplasms ; metabolism

OBJECTIVETo investigate the change of gastric cancer cell proliferation and the expression of gastric cancer related gene 213 (GCRG213), a long interspersed nuclear element-1 (LINE-1) endonuclease variant, during hypoxia.

METHODSNormal gastric mucosa cell GES-1 and gastric cancer cell BGC-823 were cultured in 20% or 3% oxygen concentrations, respectively. MTT test was used to analyze the proliferation of the GES-1 and BGC-823 cells. The change of GCRG213 mRNA and protein expression in GES-1 and BGC-823 cells was detected by using RT-PCR and Western blot analysis. Blast was used at the NCBI Blast server to identify GCRG213 sequence to any alignment in the GeneBank databases.

RESULTSCompared with 20% oxygen condition, 3% oxygen concentration could promote cell growth. Mean-while, the expression of GCRG213 at mRNA and protein levels was increased. GCRG213 sequence shared high homology with LINE-1 endonuclease sequence.

CONCLUSIONGCRG213 is a variant of LINE-1 endonuclease. Hypoxia as in 3% oxygen condition can promote cell proliferation and lead to GCRG213 overexpression.

Cell Line, Tumor ; Cell Proliferation ; Cells, Cultured ; Deoxyribonuclease I ; genetics ; Gastric Mucosa ; cytology ; Gene Expression ; Humans ; Hypoxia ; Peptide Hormones ; genetics ; Stomach Neoplasms ; genetics ; pathology

AIMTo study the influence of acetylcholine (ACh) on nicotinic receptor(N receptor) beta-subunit of the gastric epithelia and the gastric adenocarcinoma cells, and the difference of both cells.

METHODSImmunohistochemistry method was used to examine the number, number density and surface density of N receptor beta-subunit in both cells cultured in vitro.

RESULTSThe number and number density of N receptor beta-subunit in the gastric adenocarcinoma cells were much more than that in the gastric epithelia (P < 0.05). But surface density of N receptor beta-subunit in the gastric adenocarcinoma cells were lower than that in the gastric epithelia (P < 0.05). ACh at 10(6) mol/L could increase the number, number density and surface density of N receptor beta-subunit in the gastric epithelia (P < 0.01). The increase effect could not be blocked by atropine. ACh had no effect on N receptor beta-subunit in the gastric adenocarcinoma cells.

CONCLUSIONACh at low concentration initiates N receptor desensitization in the gastric epithelia. ACh has no effect on sensitivity of N receptor beta-subunit in the gastric adenocarcinoma cells.

Acetylcholine ; pharmacology ; Adenocarcinoma ; metabolism ; Cells, Cultured ; Epithelial Cells ; drug effects ; Gastric Mucosa ; cytology ; Humans ; Receptors, Nicotinic ; metabolism ; Stomach Neoplasms ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo evaluate the effect of IL-1beta on the expression of CDX2 in human gastric epithelial cell line GES-1 and its role in the intestinal metaplasia.

METHODSGES-1 cells were treated with IL-1beta in different concentrations and the expressions of CDX2 mRNA and protein were detected by real-time PCR, immunocytochemistry and Western blot at different time points. GES-1 cells were then pre-treated with NF-KappaB pathway inhibitor PDTC, and the expression of CDX2 mRNA and protein induced by IL-1beta were detected. The cell ultra-structure of GES-1 cells was observed by electronic microscope after GES-1 being treated with IL-1beta for 25 days.

RESULTSLevels of CDX2 mRNA and protein were 0.0749 + or - 0.0021 and 0.56 + or - 0.04 in the cells treated with 1 microg/L IL-1beta(P<0.05). After pre-treatment with PDTC, levels of CDX2 mRNA and protein were 0.0006 + or - 0.0002 and 0.40 + or - 0.06(P<0.05). Some changes in the cell ultra-structure of GES-1 were found by electronic microscope when GES-1 was treated with IL-1beta for 25 days.

CONCLUSIONIL-1beta can stimulate CDX2 mRNA and protein expression in GES-1 cells through the NF-KappaB signal pathway, indicating that IL-1beta plays an important role in the intestinal metaplasia.

CDX2 Transcription Factor ; Cell Line ; Epithelium ; metabolism ; pathology ; Gastric Mucosa ; cytology ; metabolism ; pathology ; Homeodomain Proteins ; metabolism ; Humans ; Interleukin-1beta ; pharmacology ; Metaplasia ; RNA, Messenger ; genetics

In the present study, rat model of gastric ischemia-reperfusion (GI-R) injury was established by clamping the celiac artery for 30 min followed by 1 h of reperfusion. Subsequently, the regulatory effect of electrical stimulation of cerebellar fastigial nucleus (FN) on GI-R injury and its neural mechanisms were investigated in Sprague-Dawley rats. The results are as follows. Electrical stimulation of the cerebellar FN not only obviously attenuated the GI-R injury in an intensity-dependent manner, but also decreased the apoptosis rate of gastric mucosal cells. Chemical lesion of FN eliminated the protective effect of electrical stimulation of FN on GI-R injury. Electrical stimulation of cerebellar FN decreased both the frequency and amplitude of the discharges of greater splanchnic nerve, but it could not change the discharge of greater splanchnic nerve following the lesion of the lateral hypothalamic area (LHA). After bilateral section of the greater splanchnic nerves, electrical stimulation of the FN also attenuated the GI-R injury. Chemical lesion of the LHA reversed the protective effect of electrical stimulation of FN on GI-R injury. Electrical stimulation of FN increased the activity of superoxide dismutase (SOD), but decreased the content of malondialdehyde (MDA) in gastric mucosa under GI-R. These results indicate that the cerebellar FN may regulate GI-R injury. Therefore, the cerebellar FN is an important brain site protecting the stomach against GI-R. The LHA and greater splanchnic nerves participate in the regulatory effects of cerebellar FN stimulation on GI-R injury. In addition, antioxidation may also be involved in the protection mechanism of cerebellar FN stimulation.
Animals ; Apoptosis ; Cerebellar Nuclei ; physiology ; Electric Stimulation ; Gastric Mucosa ; cytology ; metabolism ; Hypothalamic Area, Lateral ; physiopathology ; Malondialdehyde ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; Superoxide Dismutase ; metabolism