Objective To evaluate the gastric microbiome in patients with chronic superficial gastritis (CSG) and intestinal metaplasia (IM) and investigate the influence of Helicobacter pylori (H. pylori) on the gastric microbiome. Methods Gastric mucosa tissue samples were collected from 54 patients with CSG and IM, and the patients were classified into the following four groups based on the state of H. pylori infection and histology: H. pylori-negative CSG (n=24), H. pylori-positive CSG (n=14), H. pylori-negative IM (n=11), and H. pylori-positive IM (n=5). The gastric microbiome was analyzed by 16S rRNA gene sequencing. Results H. pylori strongly influenced the bacterial abundance and diversity regardless of CSG and IM. In H. pylori-positive subjects, the bacterial abundance and diversity were significantly lower than in H. pylori-negative subjects. The H. pylori-negative groups had similar bacterial composition and bacterial abundance. The H. pylori-positive groups also had similar bacterial composition but different bacterial relative abundance. The relative abundance of Neisseria, Streptococcus, Rothia, and Veillonella were richer in the I-HP group than in G-HP group, especially Neisseria (t=175.1, P<0.001). Conclusions The gastric microbial abundance and diversity are lower in H. pylori- infected patients regardless of CSG and IM. Compared to H. pylori-positive CSG group and H. pylori-positive IM, the relative abundance of Neisseria, Streptococcus, Rothia, and Veillonella is higher in H. pylori-positive patients with IM than in H. pylori-positive patients with CSG, especially Neisseria.
Gastric Mucosa/microbiology* ; Gastritis, Atrophic/microbiology* ; Gastrointestinal Microbiome/genetics* ; Helicobacter Infections/microbiology* ; Helicobacter pylori/genetics* ; Humans ; Metaplasia ; RNA, Ribosomal, 16S/genetics* ; Stomach Neoplasms

Proton pump inhibitors (PPIs) are commonly used to lessen symptoms in patients with gastroesophageal reflux disease (GERD). However, the effects of PPI therapy on the gastrointestinal microbiota in GERD patients remain unclear. We examined the association between the PPI usage and the microbiota present in gastric mucosal and fecal samples from GERD patients and healthy controls (HCs) using 16S rRNA gene sequencing. GERD patients taking PPIs were further divided into short-term and long-term PPI user groups. We showed that PPI administration lowered the relative bacterial diversity of the gastric microbiota in GERD patients. Compared to the non-PPI-user and HC groups, higher abundances of Planococcaceae, Oxalobacteraceae, and Sphingomonadaceae were found in the gastric microbiota from the PPI-user group. In addition, the Methylophilus genus was more highly abundant in the long-term PPI user group than in the short-term PPI-user group. Despite the absence of differences in alpha diversity, there were significant differences in the fecal bacterial composition of between GERD patients taking PPIs and those not taking PPIs. There was a higher abundance of Streptococcaceae, Veillonellaceae, Acidaminococcaceae, Micrococcaceae, and Flavobacteriaceae present in the fecal microbiota from the PPI-user group than those from the non-PPI-user and HC groups. Additionally, a significantly higher abundance of Ruminococcus was found in GERD patients on long-term PPI medication than that on short-term PPI medication. Our study indicates that PPI administration in patients with GERD has a significant effect on the abundance and structure of the gastric mucosal microbiota but only on the composition of the fecal microbiota.
Adult ; Aged ; Bacteria ; genetics ; isolation & purification ; Feces ; microbiology ; Female ; Gastric Mucosa ; microbiology ; Gastroesophageal Reflux ; drug therapy ; microbiology ; Gastrointestinal Microbiome ; drug effects ; Humans ; Male ; Microbiota ; Middle Aged ; Proton Pump Inhibitors ; therapeutic use ; RNA, Ribosomal, 16S ; genetics

BACKGROUND/AIMS: The interaction between nonsteroidal anti-inflammatory drugs (NSAIDs) and Helicobacter pylori remains controversial. We retrospectively investigated whether H. pylori infection exacerbates severe gastric mucosal injury among chronic NSAID users. METHODS: From January 2010 to December 2013, a total of 245 long-term NSAID (including low-dose aspirin) users who had undergone an esophagogastroduodenoscopy and had been evaluated for H. pylori infection were enrolled at Okayama University Hospital and Tsuyama Chuo Hospital. The degree of gastric mucosal injury was assessed according to the modified Lanza score (MLS). Severe gastric mucosal injury was defined as an MLS > or =4. Univariate and multivariate logistic regression analyses were performed. RESULTS: In the univariate analysis, age > or =75 years (odds ratio [OR], 2.4; 95% confidence interval [CI], 1.3 to 4.2), H. pylori-positivity (OR, 2.0; 95% CI, 1.2 to 3.5), and the concomitant use of proton pump inhibitors (PPIs) (OR, 0.48; 95% CI, 0.26 to 0.86) were significantly associated with severe gastric mucosal injury. The multivariate analysis was adjusted by age and sex and demonstrated that H. pylori-positivity (OR, 1.8; 95% CI, 1.0 to 3.3) and the concomitant use of PPIs (OR, 0.53; 95% CI, 0.28 to 0.99) significantly contributed to severe gastric mucosal injury. CONCLUSIONS: H. pylori infection exacerbates severe gastric mucosal injury among chronic NSAID users.
Aged ; Anti-Inflammatory Agents, Non-Steroidal/*adverse effects ; Case-Control Studies ; *Disease Progression ; Endoscopy, Digestive System ; Female ; Gastric Mucosa/*drug effects/*microbiology ; Helicobacter Infections/*complications/microbiology/pathology ; *Helicobacter pylori/drug effects ; Humans ; Logistic Models ; Male ; Middle Aged ; Odds Ratio ; Proton Pump Inhibitors/adverse effects ; Retrospective Studies

It is often difficult to differentiate gastric mucosa-associated lymphoid tissue (MALT) lymphoma from Helicobacter pylori-associated follicular gastritis, and thus, it becomes unclear how to manage these diseases. This study aimed to explore the management strategy for and the long-term outcomes of suspicious gastric MALT lymphoma detected by forceps biopsy during screening upper endoscopy. Between October 2003 and May 2013, consecutive subjects who were diagnosed with suspicious gastric MALT lymphomas by screening endoscopy in a health checkup program in Korea were retrospectively enrolled. Suspicious MALT lymphoma was defined as a Wotherspoon score of 3 or 4 upon pathological evaluation of the biopsy specimen. Of 105,164 subjects who underwent screening endoscopies, 49 patients with suspicious MALT lymphomas who underwent subsequent endoscopy were enrolled. Eight patients received a subsequent endoscopy without H. pylori eradication (subsequent endoscopy only group), and 41 patients received H. pylori eradication first followed by endoscopy (eradication first group). MALT lymphoma development was significantly lower in the eradication first group (2/41, 4.9%) than in the subsequent endoscopy only group (3/8, 37.5%, P = 0.026). Notably, among 35 patients with successful H. pylori eradication, there was only one MALT lymphoma patient (2.9%) in whom complete remission was achieved, and there was no recurrence during a median 45 months of endoscopic follow-up. H. pylori eradication with subsequent endoscopy would be a practical management option for suspicious MALT lymphoma detected in a forceps biopsy specimen obtained during screening upper endoscopy.
Adult ; Aged ; Anti-Bacterial Agents/therapeutic use ; Biopsy ; Female ; Follow-Up Studies ; Gastric Mucosa/*pathology ; Gastritis/diagnosis/etiology/microbiology ; Gastroscopy ; Helicobacter Infections/complications/*diagnosis/drug therapy ; Humans ; Lymphoma, B-Cell, Marginal Zone/complications/*diagnosis/pathology ; Male ; Middle Aged ; Republic of Korea ; Retrospective Studies

Gastric dysplasia is a neoplastic lesion and a precursor of gastric cancer. The Padova, Vienna, and World Health Organization classifications were developed to overcome the discrepancies between Western and Japanese pathologic diagnoses and to provide a universally accepted classification of gastric epithelial neoplasia. At present, the natural history of gastric dysplasia is unclear. Much evidence suggests that patients with high-grade dysplasia are at high risk of progression to carcinoma or synchronous carcinoma. Therefore, endoscopic resection is required. Although patients with low-grade dysplasia have been reported to be at low risk of progression to carcinoma, due to the marked histologic discrepancies between forceps biopsy and endoscopic specimens, endoscopic resection for this lesion is recommended, particularly in the presence of other risk factors (large size; depressed gross type; surface erythema, unevenness, ulcer, or erosion; and tubulovillous or villous histology). Helicobacter pylori eradication in patients with dysplasia after endoscopic resection appear to reduce the incidence of metachronous lesions.
Anti-Bacterial Agents/therapeutic use ; Biopsy ; Carcinoma in Situ/classification/microbiology/*pathology/*surgery ; Disease Progression ; *Gastrectomy/adverse effects/methods ; Gastric Mucosa/microbiology/*pathology/*surgery ; Gastroscopy ; Helicobacter Infections/drug therapy/microbiology ; Helicobacter pylori/drug effects ; Humans ; Neoplasm Grading ; Precancerous Conditions/classification/microbiology/*pathology/*surgery ; Predictive Value of Tests ; Risk Factors ; Stomach Neoplasms/classification/microbiology/*pathology/*surgery ; Treatment Outcome

The epithelial cytokine response, associated with reactive oxygen species (ROS), is important in Helicobacter pylori (H. pylori)-induced inflammation. H. pylori induces the production of ROS, which may be involved in the activation of mitogen-activated protein kinases (MAPK), janus kinase/signal transducers and activators of transcription (Jak/Stat), and oxidant-sensitive transcription factor, nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappaB), and thus, expression of interleukin-8 (IL-8) in gastric epithelial cells. alpha-lipoic acid, a naturally occurring thiol compound, is a potential antioxidant. It shows beneficial effects in treatment of oxidant-associated diseases including diabetes. The present study is purposed to investigate whether alpha-lipoic acid inhibits expression of inflammatory cytokine IL-8 by suppressing activation of MAPK, Jak/Stat, and NF-kappaB in H. pylori-infected gastric epithelial cells. Gastric epithelial AGS cells were pretreated with or without alpha-lipoic acid for 2 h and infected with H. pylori in a Korean isolate (HP99) at a ratio of 300:1. IL-8 mRNA expression was analyzed by RT-PCR analysis. IL-8 levels in the medium were determined by enzyme-linked immunosorbent assay. NF-kappaB-DNA binding activity was determined by electrophoretic mobility shift assay. Phospho-specific and total forms of MAPK and Jak/Stat were assessed by Western blot analysis. ROS levels were determined using dichlorofluorescein fluorescence. As a result, H. pylori induced increases in ROS levels, mRNA, and protein levels of IL-8, as well as the activation of MAPK [extracellular signal-regulated kinase 1/2 (ERK1/2), c-Jun NH2-terminal kinase 1/2 (JNK1/2), p38], Jak/Stat (Jak1/2, Stat3), and NF-kappaB in AGS cells, which was inhibited by alpha-lipoic acid. In conclusion, alpha-lipoic acid may be beneficial for prevention and/or treatment of H. pylori infection-associated gastric inflammation.
Enzyme-Linked Immunosorbent Assay ; Epithelial Cells/metabolism ; Gastric Mucosa/*drug effects/metabolism/microbiology ; Gene Expression Regulation, Bacterial ; Helicobacter Infections/immunology/*metabolism ; Helicobacter pylori/drug effects/*pathogenicity ; Humans ; Interleukin-8/genetics/*metabolism ; JNK Mitogen-Activated Protein Kinases ; Janus Kinase 1 ; Mitogen-Activated Protein Kinases/*biosynthesis ; NF-kappa B/*metabolism ; RNA, Messenger/isolation & purification/metabolism ; Reactive Oxygen Species/metabolism ; STAT3 Transcription Factor ; Stomach/metabolism/*microbiology ; Thioctic Acid/*pharmacology

NADPH oxidase produces a large amount of reactive oxygen species (ROS) in Helicobacter pylori (H. pylori)-induced gastric epithelial cells. Even though ROS mediate apoptotic cell death, direct involvement of NADPH oxidase on H. pylori-induced apoptosis remains unclear. Besides, H. pylori isolates show a high degree of genetic variability. The predominant genotype of H. pylori in Korea has been reported as cagA+, vacA s1b, m2, iceA genotype. Present study aims to investigate whether NADPH oxidase-generated ROS mediate apoptosis in human gastric epithelial AGS cells infected with H. pylori in a Korean isolate. AGS cells were pretreated with or without an NADPH oxidase inhibitor diphenyleneiodonium (DPI) and cultured in the presence of H. pylori at a bacterium/cell ratio of 300:1. Cell viability, hydrogen peroxide level, DNA fragmentation, and protein levels of p53, Bcl-2, and Bax were determined. Results showed that H. pylori inhibited cell viability with the density of H. pylori added to the cells. Inhibition of NADPH oxidase by DPI suppressed H. pylori-induced cell death, increased hydrogen peroxide, DNA fragmentation, and the ratio of Bax/Bcl-2, and p53 induction in AGS cells dose-dependently. The results suggest that targeting NADPH oxidase may prevent the development of gastric inflammation associated with H. pylori infection by suppressing abnormal apoptotic cell death of gastric epithelial cells.
Apoptosis ; Apoptosis Regulatory Proteins/metabolism ; Cell Survival ; Epithelial Cells/metabolism/microbiology ; Gastric Mucosa/metabolism ; Helicobacter Infections/*metabolism/microbiology ; Helicobacter pylori/drug effects/genetics/*isolation & purification ; Humans ; NADPH Oxidase/metabolism ; Onium Compounds/*antagonists & inhibitors/pharmacology ; Oxidative Stress/drug effects ; Reactive Oxygen Species/metabolism ; Republic of Korea ; Stomach/cytology/*metabolism/microbiology

OBJECTIVETo observe the effect of Yiqi Huoxue Qingre Huashi Recipe (YHQHR, a recipe capable of supplementing qi, activating blood, clearing heat, and dissipating dampness) on ulcer healing and Helicobacter pylori (Hp) eradication rate in Hp positive peptic ulcer patients, and to explore coccoid Hp occurrence in the eradication.

METHODSTotally 80 Hp positive peptic ulcer patients were assigned to the treatment group and the control groups by random digit table, 40 in each group. All patients received standard triple therapy of Western medicine for 2 successive weeks. Those in the control group additionally took omeprazole enteric coated tablet, 20 mg each time, once per day for 4 successive weeks. Those in the treatment group additionally took YHQHR, twice per day for 6 successive weeks. The ulcer healing was observed and recorded by gastroscope after discontinued medication of 14 days. The effective rate of ulcer healing under endoscope was statistically calculated. Rapid urease test (RUT) was performed in one small piece of tissue from corpora ventriculi and sinuses ventriculi using 14C breathe test (UBT). Gastric juice was collected from the stomach. Hp urease gene amplification test (urea A-PCR) was performed in living tissue from gastric antrum. Results obtained from the above three test methods were recorded and assessed to decide the final eradiation rate. Gastric mucosa tissue was observed under electron microscope,attempting to find non-eradicated Hp, which was further observed.

RESULTSThe total curative effect under gastroscope was 97.5% (39/40 cases) in the treatment group, obviously higher than that in the control group (80.0%, 32/40 cases) (P < 0.05). The eradication rate of Hp was 75.0% (30/40 cases), obviously better than that of the control group (52.5%, 21/40 cases) (P < 0.05). The total positive Hp numbers after treatment was 14C UBT (12), RUT (8), and urea A-PCR (27), respectively. The Hp positive rate detected by 14C UBT and RUT was lower than the Hp positive rate detected by urea A-PCR (P < 0.05). Rod-like and coccoid Hp bacteria could be observed under electron microscope.

CONCLUSIONYHQHR combined standard triple therapy was more effective than standard triple therapy alone in promoting ulcer healing and elevating the eradication rate of Hp.

Breath Tests ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; Helicobacter Infections ; drug therapy ; Helicobacter pylori ; Humans ; Omeprazole ; Peptic Ulcer ; drug therapy ; microbiology ; Urea

BACKGROUND/AIMS: Helicobacter pylori colonizes on the apical surface of gastric surface mucosal cells and the surface mucous gel layer. Pronase is a premedication enzyme for endoscopy that can disrupt the gastric mucus layer. We evaluated the additive effects of pronase combined with standard triple therapy for H. pylori eradication. METHODS: This prospective, single-blinded, randomized, controlled study was conducted between June and October 2012. A total of 116 patients with H. pylori infection were enrolled in the study (n=112 patients, excluding four patients who failed to meet the inclusion criteria) and were assigned to receive either the standard triple therapy, which consists of a proton pump inhibitor with amoxicillin and clarithromycin twice a day for 7 days (PAC), or pronase (20,000 tyrosine units) combined with the standard triple therapy twice a day for 7 days (PACE). RESULTS: In the intention-to-treat analysis, the eradication rates of PAC versus PACE were 76.4% versus 56.1% (p=0.029). In the per-protocol analysis, the eradication rates were 87.5% versus 68.1% (p=0.027). There were no significant differences concerning adverse reactions between the two groups. CONCLUSIONS: According to the interim analysis of the trial, pronase does not have an additive effect on the eradication of H. pylori infection (ClinicalTrial.gov: NCT01645761).
Adult ; Amoxicillin/therapeutic use ; Anti-Bacterial Agents/*therapeutic use ; Clarithromycin/therapeutic use ; Drug Administration Schedule ; Drug Therapy, Combination/methods ; Female ; Gastric Mucosa/drug effects/microbiology ; Helicobacter Infections/*drug therapy ; Helicobacter pylori/*drug effects ; Humans ; Male ; Middle Aged ; Pronase/*therapeutic use ; Prospective Studies ; Proton Pump Inhibitors/*therapeutic use ; Single-Blind Method ; Treatment Outcome

OBJECTIVETo understand the junction protein expression of gastric mucosa including occlusal proteins (occludin), closed protein-4 (claudin-4), zonula occluden-1(ZO-1), epithelial cadherin (E-cadherin), and β ring protein (β-catenin) and the clinical significance in children with Helicobacter pylori (Hp) infection.

METHODSeventy patients in whom gastric endoscopy was performed because of nausea, vomiting, abdominal pain, bloating, acid reflux, melena, and other gastrointestinal symptoms were enrolled in this study from Dec. 2010 to Apr. 2013 in our hospital. Informed consent was signed by their parents, and the study was in accordance with the principles of medical ethics. Hp positivity was confirmed if both respiratory urea test (RUT) and Hp were positive by gastric mucosal pathology. Gastric mucosal samples from 70 patients were enrolled in this study, 23 of them were Hp negative, 47 of them were Hp positive (24 cases without peptic ulcer, 23 cases with peptic ulcer). The mRNA levels and protein expression of tight junction protein of gastric mucosa were measured by RT-PCR and Western blot respectively. The location and semi quantitative content of E-cadherin and β-catenin in gastric mucosa were detected by immunohistochemical staining method.

RESULTThe mRNA level of E-cadherin, β-catenin, ZO-1 in the Hp positive group regardless of peptic ulcer was significantly lower than that in the Hp negative group. Hp positive without peptic ulcer group were 0.0008, 0.0040, 0.0014, respectively; Hp positive with peptic ulcer group were 0.0010, 0.0090, 0.0013, respectively; Hp negative group were 0.0137, 0.0423, 0.0198, respectively (F values were 36.956, 39.893, 38.962, respectively, all P<0.05). The expression of claudin-4 mRNA in Hp positive group with peptic ulcer increased significantly, the difference among Hp positive group with peptic ulcer, Hp positive group without peptic ulcer and Hp negative group was statistically significant (0.1438 vs. 0.0926 vs. 0.0789) (F value was 11.964, P<0.05), while the difference of occludin mRNA levels among the three groups was not statistically significant.Immunohistochemistry results showed that the score of E-cadherin, β-catenin positive cell in the Hp positive patients were also significantly lower than that in the Hp negative group (t values were 3.981 and 2.340, all P<0.05, respectively). Western blot results showed that the protein levels of β-catenin in Hp positive group with peptic ulcer were significantly lower than that in Hp negative group, while the protein levels of E-cadherin in Hp positive patients regardless of peptic ulcer were decreased significantly in Hp negative group.

CONCLUSIONOur results revealed that the tight junction protein E-cadherin, β-catenin, ZO-1 expression of gastric mucosa were decreased in children with Hp infection, while claudin-4 expression was increased in Hp positive patients with peptic ulcer, suggesting that damage to gastric epithelial barrier function may be the main pathogenesis of Hp associated gastric diseases in children.

Blotting, Western ; Cadherins ; metabolism ; Child ; Claudin-4 ; metabolism ; Gastric Mucosa ; metabolism ; pathology ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Immunohistochemistry ; Occludin ; metabolism ; Peptic Ulcer ; metabolism ; microbiology ; RNA, Messenger ; Tight Junction Proteins ; metabolism ; Tight Junctions ; metabolism ; Zonula Occludens-1 Protein ; metabolism ; beta Catenin ; metabolism