Xu C

Comparative genomic study of gastric epithelial cells co-cultured with Helicobacter pylori

AIM To identify genes potentially involved in Helicobacter pylori (H. pylori)-induced gastric carcinogenesis. METHODS GES-1 cells were co-cultured with H. pylori strains isolated from patients with gastric carcinoma (GC, n = 10) or chronic gastritis (CG, n = 10) for in vitro proliferation and apoptosis assays to identify the most and least virulent strains. These two strains were cagA-genotyped and used for further in vivo carcinogenic virulence assays by infecting Mongolian gerbils for 52 wk, respectively; a broth free of H. pylori was lavaged as control. Genomic profiles of GES-1 cells co-cultured with the most and least virulent strains were determined by microarray analysis. The most differentially expressed genes were further verified using quantitative real-time polymerase chain reaction in GES-1 cells infected with the most and least virulent strains, and by immunohistochemistry in H. pylori positive CG, precancerous diseases, and GC biopsy specimens in an independent experiment. RESULTS GC-derived H. pylori strains induced a potent proliferative effect in GES-1 cells in co-culture, whereas CG-derived strains did not. The most (from a GC patient) and least (from a CG patient) virulent strains were cagA-positive and negative, respectively. At week 52, CG, atrophy, metaplasia, dysplasia, and GC were observed in 90.0%, 80.0%, 80.0%, 90%, and 60.0%, respectively, of the animals lavaged with the most virulent strain. However, only mild CG was observed in 90% of the animals lavaged with the least virulent strain. On microarray analysis, 800 differentially expressed genes (49 up- and 751 down-regulated), involving those associated with cell cycle regulation, cell apoptosis, cytoskeleton, immune response, and substance and energy metabolisms, were identified in cells co-cultured with the most virulent strain as compared with those co-cultured with the least virulent strain. The six most differentially expressed genes (with a betweenness centrality of 0.1-0.2) were identified among the significant differential gene profile network, including JUN, KRAS, BRCA1, SMAD2, TRAF1, and HDAC6. Quantitative real-time polymerase chain reaction analyses verified that HDAC6 and TRFA1 mRNA expressions were significantly more up-regulated in GES-1 cells co-cultured with the most virulent strain than in those co-cultured with the least virulent strain. Immunohistochemistry of gastric mucosal specimens from H. pylori-positive patients with CG, intestinal metaplasia (IM), dysplasia, and GC showed that moderately positive and strongly positive HDAC6 expression was detected in 21.7% of CG patients, 30.0% of IM patients, 54.5% of dysplasia patients, and 77.8% of GC patients (P < 0.001). The up-regulation of TRAF1 expressions was detected in 34.8%, 53.3%, 72.7%, and 88.9% specimens of CG, IM, dysplasia, and GC, respectively (P < 0.001). CONCLUSION The overexpression of HDAC6 and TRAF1 in GES-1 cells co-cultured with the GC-derived strain and in H. pylori-positive dysplasia and GC suggests that HDAC6 and TRAF1 may be involved in H. pylori-induced gastric carcinogenesis.

Association between Virulence Factors and TRAF1/4-1BB/Bcl-xL Expression in Gastric Mucosa Infected with Helicobacter pylori

Objective. CagA+/vacAs1+/vacAm1+ Helicobacter pylori upregulates the expression of tumor necrosis factor receptor–associated factor 1 (TRAF1), tumor necrosis factor receptor superfamily member 9 (4-1BB), and B-cell lymphoma-extra large (Bcl-xL) in human gastric epithelial cells. We investigated the correlation between cagA/vacAs1/vacAm1 and TRAF1/4-1BB/Bcl-xL expression in gastric mucosal tissue of patients with gastric disorders. Methods. We collected gastric mucosa samples from 35 chronic, nonatrophic gastritis (CG) patients, 41 atrophic gastritis patients, 44 intestinal metaplasia with atypical hyperplasia (IM) patients, and 28 gastric carcinoma (Ca) patients. The expression of  TRAF1, 4-1BB, and Bcl-xL was determined using western blotting. The expression of cagA, vacAs1, and vacAm1 in H. pylori was examined with polymerase chain reaction. Results. The expression of TRAF1, 4-1BB, and Bcl-xL was significantly upregulated in IM and Ca patients (P < 0.05 compared with CG). There were more cases of cagA+/vacAs1+/vacAm1+ H. pylori infection in samples with elevated TRAF1, 4-1BB, or Bcl-xL expression (P < 0.05). Additionally, there were a remarkably large number of samples with upregulated TRAF1/4-1BB/Bcl-xL expression in cases of cagA+/vacAs1+/vacAm1+ H. pylori infection (44 cases, 67.7%; P < 0.05). Conclusions. The pathogenesis of IM and Ca may be promoted by cagA+/vacAs1+/vacAm1+ H. pylori, possibly via upregulated TRAF1, 4-1BB, and Bcl-xL in gastric mucosal tissue.

The expression level of TRAF1 in human gastric mucosa is related to virulence genotypes of Helicobacter pylori

Abstract Objective. To investigate the expression level of tumor necrosis factor receptor-associated factor 1 (TRAF1) in gastric mucosa tissue in patients infected with Helicobacter pylori (H. pylori) and to analyze the relationship between TRAF1 expression and H. pylori virulence. Methods. Gastric tissue samples were collected from patients with gastritis, atrophic gastritis, intestinal metaplasia with atypical hyperplasia, and gastric cancer. The expression level of TRAF1 in each group was analyzed by real-time polymerase chain reaction (PCR) and Western blot analysis. Virulence genotypes of H. pylori were determined by PCR. Results. Significant differences in TRAF1 mRNA levels were observed between the gastritis and gastric cancer groups, and the atrophic gastritis and gastric cancer groups (p < 0.05). Moreover, significant differences in TRAF1 protein levels were observed between the gastritis and intestinal metaplasia with atypical hyperplasia groups, between the gastritis and gastric cancer groups, and between the atrophic gastritis and gastric cancer groups (all p < 0.05). The virulence genotypes of cytotoxin-associated gene A (cagA), vacAs1, and vacAm1 were more frequent in the TRAF1 high-level group than in the TRAF1 low-level group (p < 0.05). Conclusion. Higher TARF1 expression level is associated with infection by CagA+/vacAs1+/m1+ virulent H. pylori strains and may promote the proliferation of gastric mucosal cells and induce gastric cancer.

Stepwise sedation for elderly patients with mild/moderate COPD during upper gastrointestinal endoscopy

AIM To investigate stepwise sedation for elderly patients with mild/moderate chronic obstructive pulmonary disease (COPD) during upper gastrointestinal (GI) endoscopy. METHODS Eighty-six elderly patients with mild/moderate COPD and 82 elderly patients without COPD scheduled for upper GI endoscopy were randomly assigned to receive one of the following two sedation methods: stepwise sedation involving three-stage administration of propofol combined with midazolam [COPD with stepwise sedation (group Cs), and non-COPD with stepwise sedation (group Ns)] or continuous sedation involving continuous administration of propofol combined with midazolam [COPD with continuous sedation (group Cc), and non-COPD with continuous sedation (group Nc)]. Saturation of peripheral oxygen (SpO2), blood pressure, and pulse rate were monitored, and patient discomfort, adverse events, drugs dosage, and recovery time were recorded. RESULTS All endoscopies were completed successfully. The occurrences of hypoxemia in groups Cs, Cc, Ns, and Nc were 4 (9.3%), 12 (27.9%), 3 (7.3%), and 5 (12.2%), respectively. The occurrence of hypoxemia in group Cs was significantly lower than that in group Cc (P < 0.05). The average decreases in value of SpO2, systolic blood pressure, and diastolic blood pressure in group Cs were significantly lower than those in group Cc. Additionally, propofol dosage and overall rate of adverse events in group Cs were lower than those in group Cc. Finally, the recovery time in group Cs was significantly shorter than that in group Cc, and that in group Ns was significantly shorter than that in group Nc (P < 0.001). CONCLUSION The stepwise sedation method is effective and safer than the continuous sedation method for elderly patients with mild/moderate COPD during upper GI endoscopy.

Effects of different types of Helicobacter pylori on the gap junction intercellular communication in GES-1 cells

OBJECTIVE To determine the effect of different types of Helicobacter pylori (H.pylori) on the gap junction intercellular communication (GJIC) in GES-1 cells, and investigate the types of H.pylori related to the dysfunction of GJIC. METHODS Different types of H.pylori clinical strains were isolated and cultured, including the East Asian CagA(-)positive H.pylori (East Asian CagA(+)H.pylori), Western CagA(-)positive H.pylori (Western CagA(+)H.pylori), and the CagA(-)negative H.pylori (CagA(-)H.pylori). We co-cultured these H.pylori strains with GES-1 cells for 24 and 48 h, respectively. The control group was cultured without any H.pylori for 24 and 48 h. Change of the GJIC function in GES-1 cells was detected by the scrape-loading dye transfer (SLDT) technique. The cell proliferation of each group was examined by the methyl thiazolyl tetrazolium bromide (MTT) assay. RESULTS The control group showed better GJIC function in the GES-1 cells, and the fluorescent dye migrated 4-5 rows to the adjacent cells at 24 and 48 h. Compared with the control group, the GJIC function of GES-1 cells in the CagA(-)H.pylori group decreased and the fluorescent dye migrated 3 rows to the adjacent cells. Compared with the control group and the CagA(-) H.pylori group, the GJIC function of GES-1 cells in the Western CagA(+)H.pylori group decreased and the fluorescent dye migrated 1-2 rows to the adjacent cells. The East Asian CagA(+)H.pylori group showed no GJIC function or weak GJIC function, and most of the fluorescent dye was confined to the area of scratched single row cells and only a few migrated 1-2 rows to the adjacent cells. Difference in the cell proliferation between the CagA(-)H.pylori group and the control group was not significant. The cell proliferation of the Western CagA(+)H.pylori group and the East Asian CagA(+)H.pylori group at bacterium-to-cell ratio of 100:1 and 200:1 was higher than that of the control group. The cell prolife-ration of the East Asian CagA(+)H.pylori group at bacterium-to-cell ratio of 400:1 was significantly lower than that of the control group at 48 h. CONCLUSION H.pylori can inhibit the GJIC function in GES-1 cells, which may be associated with CagA(+)H.pylori, especially with East Asian CagA(+)H.pylori. The effect of H.pylori on the proliferation of GES-1 cells is related to virulence factor CagA.

Effect of birid triple viable on peptic ulcer patients with Helicobacter pylori infection

OBJECTIVE To observe the effect of birid triple viable on peptic ulcer patients with Helicobacter pylori(H.pylori) infection. METHODS A total of 120 peptic ulcer patients with H.pylori infection was randomly divided into 2 groups. The control group was treated with the triple therapy including Esomeprazole, Amoxicillin, and Furazolidone. The treatment group was treated with the same triple therapy plus birid triple viable. The pH values of the gastric juice, bacterial culture of the gastric juice, the therapy-related side effects and the healing rate of ulcer and erosive were observed before the treatment and after a 14-day-treatment respectively. The (14)C-urea breath test((14)C-UBT) was used to evaluate the H.pylori eradication rates 4 weeks after the treatment. RESULTS After the treatment, pH values of the gastric juice in both groups were significantly higher than those before the treatment (P<0.05). When pH values of the gastric juice ≥4, the positive rate of bacterial culture in the treatment group was lower than that of the control group (80.0% and 97.4% respectively, P<0.05). The rate of adverse effects in the treatment group was lower than that in the control group (11.7% and 30% respectively, P<0.05).After a 2-week treatment, the healing rate of ulcer and erosion was 80% and 68.7% in the treatment group, and 73.3% and 40.7% in the control group. The healing rate of ulcer had no significant difference in the 2 groups (P>0.05),while the healing rate of erosion in the treatment group was higher than that of the control group (P<0.05).The H.pylori eradication rates evaluated by per-protocol (PP) analysis in the treatment group was significantly higher than that of the control group (80.4% and 62.4% respectively, P<0.05). CONCLUSION The combined use of birid triple viable and the triple therapy of H.pylori in peptic ulcer patients with H.pylori infection can reduce the bacteria of the gastric juice and therapy-related side effects. It can increase the H.pylori eradication rate and promote the healing of erosion.

Correlation between the cystathionine-r-lyase (CES) and the severity of peptic ulcer disease

BACKGROUND The infection of Helicobacter pylori (H. pylori) is one of the most important causes of gastric ulcer disease. The role of hydrogen sulfide (H2S) production in H. pylori-induced gastric ulcer disease. AIM The expression of cystathionine-γ-lyase (CSE) was determined, and correlated with the severity of gastric ulcer disease. METHODS One hundred and eight patients were selected based on the determination of gastric ulcer and the infection of Helicobacter pylori (H. pylori), including 36 normal control, 36 patients with H. Pylori-negative gastric ulcer, and 36 patients with H. Pylori-positive gastric ulcer. RT-PCR determination was performed to determine the expression of CSE, NF-κB and IL-8. RESULTS The expression of CSE, NF-κB and IL-8 was higher in the gastric ulcer group than control group (p<0.05). Compared with the H. pylori-negative gastric ulcer, the expression of CSE, NF-κB and IL-8 was higher than H. pylori-positive gastric ulcer group (p<0.05). For H. pylori-negative gastric ulcer group, the expression of CSE positively correlated with the expression of NF-κB (r=0.98, p<0.05) and IL-8 (r=0.95, p<0.05). For H. pylori-positive gastric ulcer group, the expression of CSE also positively correlated with the expression of NF-κB (r=0.99, p<0.05) and IL-8 (r=0.85, p<0.05). CONCLUSION The expression of CSE was positively correlated with the severity of gastric ulcer.

Relationship between the chronic periodontitis and the depression anxiety psychological factor

OBJECTIVE To explore the relationship between the chronic periodontitis (CP) and the depression-anxiety psychological factors. METHODS Thirty-one patients and 29 age, gender-matched volunteers were enrolled for this study. In order to assess the depression-anxiety psychological index, the subjects filled the questionnaire regarding the demographic and socioeconomic information, the oral hygiene habit, the Center for Epidemiologic Studies Depression Scale (CES-D) and the Self Rating Anxiety Scale(ASA). Calculus index (CI), bleeding on probing (BOP), probing depth (PD), clinical attachment loss (CAL), furcation involvement (FI) and tooth mobility were assessed at 6 sites per tooth of all erupted teeth by a manual periodontal probe. The data were analyzed by the analysis of variance, χ(2) test, and multivariable logistic step wise analysis via the software of SPSS 15.0. RESULTS The mean CAL of the control group was 0.46 ± 0.16,the mean CAL of the moderate, high, and severe CP group was 2.84 ± 0.12, 3.51 ± 0.34, and 4.71 ± 0.51, respectively, which is significant difference between each other (P<0.01). The depression index of the volunteers, the moderate CP, the high CP, and the severe CP was 30.52 ± 3.73, 35.83 ± 7.76, 37.25 ± 6.16, 37.82 ± 5.94, respectively. The anxiety index among the 4 groups was 26.69 ± 3.55, 37.67 ± 6.31, 32.87 ± 5.54, and 35.94 ± 6.30, respectively. The depression and anxiety indexes of the periodontitis groups were higher than those of the control (P<0.01) while there was no significant difference among the 3 CP groups (P>0.05). The multivariate logistic analysis of the relationship between CP and the depression-anxiety psychological factors showed that the depression psychological factor was B=2.301,OR=9.988 while the optimistic coping style was B=-5.174,OR=0.006 in the equation of the regression. CONCLUSION The depression psychological factor was related to the progression of CP. In addition, the optimistic coping style could prevent the progression of the CP.

Chronic Helicobacter pylori infection induces the proliferation and apoptosis in gastric epithelial cells and gastric precancerosis in Mongolian gerbils

OBJECTIVE To explore the effect of different Helicobacter pylori (H.pylori) clinical strains on the proliferation and apoptosis of gastric epithelial cells, and to observe the effect of H.pylori on gastric mucosa by Mongolian gerbil model infected H.pylori. METHODS H.pylori isolates harvested from pathologically documented gastric carcinoma (GC, n=10) or chronic gastritis specimens (CG, n=10) were co-cultured with GES-1 cells individually. MTT assay and flow cytometry were used to determine the proliferation and apoptosis of GES-1 cells induced by H.pylori isolates. Mongolian gerbils were infected by the most (A strain) and the least (B strain) significantly proliferated H.pylori strains. Results When co-cultured with the cell/bacteria concentration ratio 1:1 and 1:50 for 12 h and the cell/bacteria concentration ratio 1:50 for 24 h, H.pylori clinical strains isolated from patients with gastric cancer promoted the proliferation of GES-1 cells, and there was significant difference in the absorbance compared with the group of gastritis strains(P<0.05). The apoptosis rate of the GC and CG groups increased significantly (P<0.05) compared with the control group when co-cultured with the cell/bacteria concentration ratio 1:50 and 1:200, and there was no significant difference between the GC group and the CG group (P>0.05). The incidences of intestinal metaplasia and dysplasia in the A strain group were significantly higher than those in the B strain group (P<0.05). CONCLUSION H.pylori strains from different disease sources have different effects on the proliferation of GES-1 cells. H.pylori isolated from gastric cancer can promote the proliferation of cells to different degrees and directly induce gastric precancerosis and gastric cancer.

PBX1 attributes as a determinant of connexin 32 downregulation in Helicobacter pylori-related gastric carcinogenesis

AIM To clarify the mechanisms of connexin 32 (Cx32) downregulation by potential transcriptional factors (TFs) in Helicobacter pylori (H. pylori)-associated gastric carcinogenesis. METHODS Approximately 25 specimens at each developmental stage of gastric carcinogenesis [non-atrophic gastritis, chronic atrophic gastritis, intestinal metaplasia, dysplasia and gastric carcinoma (GC)] with H. pylori infection [H. pylori (+)] and 25 normal gastric mucosa (NGM) without H. pylori infection [H. pylori (-)] were collected. After transcriptional factor array analysis, the Cx32 and PBX1 expression levels of H. pylori-infected tissues from the developmental stages of GC and NGM with no H. pylori infection were measured by real-time polymerase chain reaction (RT-PCR) and Western blot analysis. Regarding H. pylori-infected animal models, the Cx32 and PBX1 mRNA expression levels and correlation between the gastric mucosa from 10 Mongolian gerbils with long-term H. pylori colonization and 10 controls were analyzed. PBX1 and Cx32 mRNA and protein levels were further studied under the H. pylori-infected condition as well as PBX1 overexpression and knockdown conditions in vitro. RESULTS Incremental PBX1 was first detected by TF microarray in H. pylori-related gastric carcinogenesis. The identical trend of PBX1 and Cx32 expression was confirmed in the developmental stages of H. pylori-related clinical specimens. The negative correlation of PBX1 and Cx32 was confirmed in H. pylori-infected Mongolian gerbils. Furthermore, decreased PBX1 expression was detected in the normal gastric epithelial cell line GES-1 with H. pylori infection. Enforced overexpression or RNAi-mediated knockdown of PBX1 contributed to the diminished or restored Cx32 expression in GES-1 and the gastric carcinoma cell line BGC823, respectively. Finally, dual-luciferase reporter assay in HEK293T cells showed that Cx32 promoter activity decreased by 30% after PBX1 vector co-transfection, indicating PBX1 as a transcriptional downregulator of Cx32 by directly binding to its promoters. CONCLUSION PBX1 is one of the determinants in the Cx32 promoter targeting site, preventing further damage of gap junction protein in H. pylori-associated gastric carcinogenesis.