OBJECTIVES: Helicobacter pylori infection induces selective reduction of the number of antral D-cells and results in abnormal regulation of serum gastrin secretion. The purpose of this study was to investigate the relationship between H. pylori infection and the numbers of G-cells and D-cells. METHODS: The numbers of antral G-cells and D-cells, the ratio of G-cells to D-cells and fasting serum gastrin concentrations were compared between 37 patients with (29 with duodenal ulcers and 8 with gastric ulcers) and 33 without H. pylori infection (22 with duodenal ulcers and 11 with gastric ulcers). Serum gastrin concentrations were measured using the radioimmunoassay technique. Antral mucosal biopsy specimens were examined using immunohistochemical staining with antibodies specific for gastrin and somatostatin and the numbers of G-cells and D-cells per gastric gland were counted. RESULTS: Fasting serum gastrin concentrations were significantly higher in patients with H. pylori infection compared to patients without infection (80.3 +/- 23.5 vs 47.6 +/- 14.1 pg/ml, p < 0.001). The number of G-cells per gastric gland was similar in infected and uninfected patients (7.1 +/- 3.1 vs 7.3 +/- 3.9, respectively, p > 0.5). The number of D-cells was significantly lower in patients with H. pylori infection than in uninfected patients in both duodenal and gastric ulcer patients (1.3 +/- 0.4 vs 2.5 +/- 1.6, respectively, p < 0.001). The ratio of G-cells to D-cells was also significantly higher in infected patients compared with uninfected patients for both gastric and duodenal ulcers (5.7 +/- 2.7 vs 3.5 +/- 1.9, respectively, p < 0.001). CONCLUSIONS: These results strongly suggest that Helicobacter pylori infection induces reduction of the number of antral D-cells. The resulting relative hypofunction of the inhibitory action of D-cells against G-cells may be responsible for increased serum gastrin secretion.
Case-Control Studies ; D Cells/pathology ; D Cells/metabolism ; G Cells/pathology ; G Cells/metabolism ; Gastrins/metabolism ; Gastrins/blood ; Gastritis/pathology ; Gastritis/metabolism* ; Helicobacter Infections/pathology ; Helicobacter Infections/metabolism* ; Helicobacter pylori* ; Human ; Somatostatin/metabolism

AIMAccumulated evidence suggest that the development of vascular calcification is similar to osteogenesis. Here we want to elucidate the effect of the common used osteo-regulatory factor 1,25(OH)2D3 on vascular calcification.

METHODS AND RESULTSAdding 10(-9) mol/L to the culture media 1,25(OH)2D3 time dependently increased the calcium deposition on the in vitro calcification of bovine vascular smooth muscle cells (BVSMCs) induced by beta-GP. It also increased cellular alkaline phosphatase activity by 301.1% during the calcified process. Osteocalcin, one of the osteogenic specific metric proteins, was dramatically elevated by 58.3% during the calcified processes, which indicate the transformation of BVSMCs to osteoblastic cell. 1,25(OH)2D3 had no such effect on non-calcified BVSMCs.

CONCLUSIONThese data suggest that 1,25(OH)2D3 exerts a stimulatory effect on vascular calcification through increasing the synthesis of ALP. This effect shares the same character as osteoblast cells. This effect is limited to the calcified prone vascular cell.

Animals ; Calcitriol ; metabolism ; Cattle ; Cells, Cultured ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; drug effects ; metabolism ; pathology ; Osteocalcin ; metabolism ; Vascular Calcification ; metabolism ; pathology ; Vitamin D ; analogs & derivatives ; pharmacology

OBJECTIVETo study the role of N-methyl-D-aspartate receptor and Ca(2+) in acute intoxicated encephalopathy induced by 1, 2-dichloroethane (1, 2-DCE) in vitro.

METHODSNeurocytes of new born rats were cultured in vitro, which were administered with different doses of 1, 2-DCE, and NMDAR and Ca(2+) antagonists including Ketamine and Nimodiping respectively. The cell morphologic structures were observed under light microscope, and its proliferation was detected by Cell Counting Kit-VIII.

RESULTS1, 2-DCE could damage the normal morphological structure of neurocytes: the cell body swelled and broke down, the karyon slurred or disappeared, the axone became shorten and thick, connection of neurocytes was reduced, the cell membrane was half-baked, injury of neurocytes became severer with the increase of the dose of 1, 2-DCE. There was no statistical difference in the proliferation of neurocytes between every 1, 2-DCE groups (P > 0.05), but there was significantly statistical difference between 1, 2-DCE groups, the control group, and the retarder groups (P < 0.01).

CONCLUSION1, 2-DCE can damage the normal morphological structure of neurocytes, and the damage will become severer with the increase of the dose of 1, 2-DCE. However, the cell morphologic structures and proliferation of antagonist groups are much better than those in the 1, 2-DCE groups.

Animals ; Calcium ; antagonists & inhibitors ; physiology ; Cells, Cultured ; Ethylene Dichlorides ; toxicity ; Neurons ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; antagonists & inhibitors ; physiology

OBJECTIVE: To investigate the effect of overexpression of glycosylphosphatidyl-inositol-specific phospholipase D (GPI-PLD) on the biological character of hepatocellular carcinoma cell line HepG2. METHODS: The GPI-PLD gene eukaryon expression vector pcDNA3.1(+)/ GPI-PLD was transiently transfected into HepG2 cell by lipid-media transfection. The untransfected HepG2 and HepG2 transfected with pcDNA3.1(+) were used as controls. After screening with G418, the single clone was obtained. The expression level of GPI-PLD mRNA in HepG2 was identified by reverse transcription polymerase chain reaction (RT-PCR). GPI-PLD activities were analyzed quantitatively by triton-X-114 partition with GPI anchored placental alkaline phosphatase (PLAP) as a substrate. Cell count was used to detect the proliferation of the 3 groups, and complement dependent cytotoxicity (CDC) effects were observed by the staining of trypan blue. Apoptosis cells were analyzed by flow cytometry. Carcinoembryonic antigen (CEA)was detected by enzyme linked immunosorbent assay (ELISA). RESULTS: Compared with HepG2 and pcDNA3.1(+)/HepG2 cell, the levels of GPI-PLD activities and its mRNA from pcDNA3.1(+)/GPI-PLD/HepG2 were increased with almost 2 to 5 times,respectively. The GPI anchored PLAP and CEA released into the medium by GPI-PLD, and the rate of CDC killing on the cells were significantly increased. However, the proliferative capacity was obviously decreased, and the typical apoptosis cells were presented in positive clones and its apoptosis rates were increased significantly. CONCLUSION The stable cell line with overexpression of GPI-PLD has been constructed. The overexpression of GPI-PLD in these cells increases the sensitivity of these cells to CDC killing and impairs the proliferative capacity of cells, and promotes the apoptosis.
Apoptosis ; genetics ; Carcinoma, Hepatocellular ; genetics ; pathology ; Complement Activation ; genetics ; Cytotoxicity, Immunologic ; genetics ; Eukaryotic Cells ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Humans ; Liver Neoplasms ; genetics ; pathology ; Phospholipase D ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Transfection ; Tumor Cells, Cultured ; Up-Regulation

The development of a serological marker for early diagnosis of isocyanate-induced occupational asthma (isocyanate-OA) may improve clinical outcome. Our previous proteomic study found that expression of vitamin D-binding protein (VDBP) was upregulated in the patients with isocyanate-OA. In the present study, we evaluated the clinical relevance of VDBP as a serological marker in screening for isocyanate-OA among exposed workers and its role in the pathogenesis of isocyanate-OA. Three study groups including 61 patients with isocyanate-OA (group I), 180 asymptomatic exposed controls (AECs, group II), 58 unexposed healthy controls (NCs, group III) were enrolled in this study. The baseline serum VDBP level was significantly higher in group I compared with groups II and III. The sensitivity and specificity for predicting the phenotype of isocyanate-OA with VDBP were 69% and 81%, respectively. The group I subjects with high VDBP (> or = 311 microg/ml) had significantly lower PC20 methacholine levels than did subjects with low VDBP. The in vitro studies showed that TDI suppressed the uptake of VDBP into RLE-6TN cells, which was mediated by the downregulation of megalin, an endocytic receptor of the 25-hydroxycholecalciferol-VDBP complex. Furthermore, toluene diisocyanate (TDI) increased VEGF production and secretion from this epithelial cells by suppression of 1,25-dihydroxycholecalciferol [1,25(OH)2D3] production. The findings of this study suggest that the serum VDBP level may be used as a serological marker for the detection of isocyanate-OA among workers exposed to isocyanate. The TDI-induced VEGF production/secretion was reversed by 1,25(OH)2D3 treatment, which may provide a potential therapeutic strategy for patients with isocyanate-OA.
Adult ; Animals ; *Asthma/blood/chemically induced/pathology ; Cell Line ; Epithelial Cells ; Female ; Gene Expression/drug effects ; Humans ; Isocyanates/toxicity ; Male ; Middle Aged ; *Occupational Diseases/blood/chemically induced/pathology ; Rats ; Toluene 2,4-Diisocyanate/toxicity ; Vitamin D-Binding Protein/*blood

The effect of zinc on the damage of primary cultured hippocampal neurons induced by corticosterone (CORT) was studied. Neuronal injury and expression of NMDA receptor subunits (NR1,NR2A,NR2B) mRNA were detected by using in situ staining and RT-PCR, respectively. Neurons treated with 5 micromol/L CORT for 24 h showed decreased survival rates and increased apoptotic rates compared with the controls; co-application of CORT and 10 or 100 micromol/L Zn(2+) attenuated apoptotic rates while 250 micromol/L Zn(2+) worsened CORT-induced neuronal injury. Expression of NR1, NR2B mRNA in neurons treated by 5 micromol/L CORT for 24 h was significantly increased, while those concurrently added with 10 or 100 micromol/L Zn(2+) showed no changes. No statistic difference in NR2A mRNA was obtained under any treatment. These results suggest that zinc can bilaterally regulate neuronal injuries induced by CORT, among while NMDA receptors probably play an important role.
Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Cells, Cultured ; Corticosterone ; pharmacology ; Hippocampus ; pathology ; Neurons ; pathology ; Neuroprotective Agents ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Receptors, N-Methyl-D-Aspartate ; biosynthesis ; classification ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Zinc ; pharmacology

OBJECTIVE: To clarify the effect of glycosylphosphatidylinositol-specific phospholipase D (GPIPLD) on hepatoma cells HepG2 and the possible molecular mechanism. METHODS: MTT, fluorescent staining and Western blot were applied to analyze the effect and molecular mechanism of GPI-PLD on hepatoma cells by transfected high expression GPI-PLD model. We inoculated HepG2 in nude mice models to further clarify the effect of GPI-PLD on hepatoma cells in vivo. RESULTS: Compared with the control groups, PI3K-Akt signaling pathway activity and proliferation of hepatoma cells were significantly inhibited in the GPI-PLD group. Nude mice models showed that the tumor growth and tumor weight [(1.87 ± 0.09) g] of the GPI-PLD group were significantly less than those of the blank control group [(2.20 ± 0.17) g] and the negative control group [(2.15 ± 0.09) g]. AST, ALT and AFP serum concentration in the GPI-PLD group were significantly lower than those of the control groups (P<0.05). CONCLUSION GPI-PLD can inhibit the proliferation of hepatoma cells and growth in vivo, and promote the apoptosis of hepatoma cells by reducing the activity of PI3K-Akt signaling pathway.
Animals ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line ; Down-Regulation ; Hep G2 Cells ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Mice ; Mice, Nude ; Phosphatidylinositol 3-Kinases ; Phospholipase D ; metabolism ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Transfection

This study was purposed to investigate the expression and significance of glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) in bone marrow mononuclear cells (BMMNC) isolated from patients with acute myeloid leukemia (AML), GPI-PLD activity in BMMNC isolated from 78 patients with AML and 15 normal persons was measured by using GPI-anchored placental alkaline phosphatase (PLAP) as a substrate and Triton X-114 phase partitioning. The GPI-PLD mRNA expression was measured by semi-quantitive reverse transcription-polymerase chain reaction (RT-PCR). The results showed that the mRNA expression level and activity of GPI-PLD in BMMNC from de novo AML patients were 1.86 +/- 0.32 and 46.96 +/- 7.15% respectively; the mRNA expression level and activity of GPI-PLD in BMMNC from completely remission and refractory or relapsed patients were 1.26 +/- 0.29, 33.36 +/- 5.13%and 1.79 +/- 0.19, 44.31 +/- 7.22%, while those in BMMNC from normal controls were 1.27 +/- 0.23, 35.38 +/- 5.15% respectively. The mRNA expression level and activity of GPI-PLD in de novo and refractory or relapsed patients were obviously higher than those in normal controls with significant difference (p < 0.01), while the comparison between remitted patients and normal controls showed no statistical difference (p > 0.05). It is concluded that the expression level of GPI-PLD mRNA coincides with GPI-PLD activity. The mRNA expression and activity of GPI-PLD in de novo and refractory or relapsed patients are obviously higher than those in normal controls. It is worthy of further exploring whether GPI-PLD plays a certain role in process of leukemia pathogenesis.
Adolescent ; Adult ; Bone Marrow Cells ; cytology ; metabolism ; Case-Control Studies ; Child ; Female ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Male ; Middle Aged ; Phospholipase D ; genetics ; metabolism ; RNA, Messenger ; genetics ; Young Adult

OBJECTIVETo investigate the dual effects by the delta opioid receptor agonists DPDPE on the delayed rectified potassium channels in NG108-15 cells.

METHODSA series of outward currents were evoked in NG108-15 cells by depolarizing voltage from -50 mV to +80 mV at holding potential of -90 mV. These currents were delayed rectified potassium currents. Relatively selected delta opioid receptor agonists DPDPE of higher and lower concentrations were used to modulate the delayed rectified K+ current in NG108-15 cells. Opioid receptor antagonist Naloxone (NAL) and relatively selected delta opioid receptor antagonist Naltrindole (NTI) were used in the present experiments for the characterization of the actions of opioid receptors.

RESULTSThe relatively higher concentrations of delta opioid receptor agonist DPDPE (> or = 10(-6) mol/L) significantly increased the amplitude of the delayed rectified K+ current. On the contrary, the relatively lower concentrations of DPDPE (< or = 10(-12) mol/L) decreased the amplitude of the delayed rectified K+ current (P < 0.05). Furthermore both the increase and decrease were time-dependent.

CONCLUSIONSdelta opioid receptor agonist has dual regulatory effects on the delayed rectified potassium channels in NG108-15 cells.

Animals ; Cell Membrane ; metabolism ; Enkephalin, D-Penicillamine (2,5)- ; pharmacology ; Glioma ; metabolism ; pathology ; Hybrid Cells ; metabolism ; Mice ; Naloxone ; pharmacology ; Naltrexone ; analogs & derivatives ; pharmacology ; Neuroblastoma ; metabolism ; pathology ; Patch-Clamp Techniques ; Potassium Channels, Inwardly Rectifying ; drug effects ; metabolism ; Rats ; Receptors, Opioid, delta ; agonists ; Tumor Cells, Cultured

OBJECTIVETo observe the role of G protein in the dual regulation of opioid receptor agonist on the delayed rectified potassium channels.

METHODSUsing whole-cell patch-clamp techniques applied to NG108-15 cells, investigate the effect of opioid receptor agonist on the delayed rectified potassium channels by administration of Guanosine-5'-0'-2-thiociphosphate (GDP beta S), Pertusis Toxin (PTX), Tetroacetic acid nueleoside diphosphate kinase (NDPK) and Adenosine-3' 5' cyclic monophosphate cAMP in the pipette solution.

RESULTS(1) GDP beta S could block the changes induced by both high and low concentration of (D-Pen2.5)-enkephalin (DPDPE) (P < 0.05). (2) PTX could inhibit the excitative regulation on K+ channel by high concentration of DPDPE (P < 0.05). But CTX had no effect on K+ channel caused by DPDPE. (3) UDP could block the excitative effect of K+ channel by high concentration of NDPK, while have no changes on the inhibitory effect caused by low concentration of opioid agonists. (4) cAMP took part in the regulation in high concentration of agonist administration (P < 0.05), while no changes for low concentration of agonists.

CONCLUSIONSDual changes were observed on delayed rectifier potassium channel by agonist treatment on NG108-15 cells. The excitative effect was Gi/o coupled in high concentration of agonist incubation, related to cAMP. While the inhibitory effect was possibly induced by G protein beta gamma subunit directly.

Animals ; Enkephalin, D-Penicillamine (2,5)- ; pharmacology ; GTP-Binding Proteins ; physiology ; Glioma ; metabolism ; pathology ; Guanosine Monophosphate ; analogs & derivatives ; pharmacokinetics ; Hybrid Cells ; metabolism ; pathology ; Mice ; Neuroblastoma ; metabolism ; pathology ; Patch-Clamp Techniques ; Pertussis Toxin ; pharmacology ; Potassium Channels, Inwardly Rectifying ; metabolism ; Rats ; Receptors, Opioid ; agonists ; Thionucleotides ; pharmacokinetics