1.Correlations between the CagA Antigen and Serum Levels of Anti-Helicobacter pylori IgG and IgA in Children.
Ji Hyun SEO ; Chun Woo LIM ; Ji Sook PARK ; Jung Sook YEOM ; Jae Young LIM ; Jin Su JUN ; Hyang Ok WOO ; Hee Shang YOUN ; Seung Chul BAIK ; Woo Kon LEE ; Myung Je CHO ; Kwang Ho RHEE
Journal of Korean Medical Science 2016;31(3):417-422
We tested correlations between anti-Helicobacter pylori IgG and IgA levels and the urease test, anti-CagA protein antibody, degree of gastritis, and age. In total, 509 children (0-15 years) were enrolled. Subjects were stratified as 0-4 years (n = 132), 5-9 years (n = 274), and 10-15 years (n = 103) and subjected to the urease test, histopathology, ELISA, and western blot using whole-cell lysates of H. pylori strain 51. The positivity rate in the urease test (P = 0.003), the degree of chronic gastritis (P = 0.021), and H. pylori infiltration (P < 0.001) increased with age. The median titer for anti-H. pylori IgG was 732.5 IU/mL at 0-4 years, 689.0 IU/mL at 5-9 years, and 966.0 IU/mL at 10-15 years (P < 0.001); the median titer for anti-H. pylori IgA was 61.0 IU/mL at 0-4 years, 63.5 IU/mL at 5-9 years, and 75.0 IU/mL at 10-15 years (P < 0.001). The CagA-positivity rate was 26.5% at 0-4 years, 36.5% at 5-9 years, and 46.6% at 10-15 years for IgG (P = 0.036), and 11.3% at 0-4 years, 18.6% at 5-9 years, and 23.3% at 10-15 years for IgA (P < 0.001). Anti-H. pylori IgG and IgA titers increased with the urease test grade, chronic gastritis degree, active gastritis, and H. pylori infiltration. Presence of CagA-positivity is well correlated with a high urease test grade and high anti-H. pylori IgG/IgA levels.
Adolescent
;
Antibodies, Bacterial/*blood
;
Antigens, Bacterial/*analysis/immunology
;
Bacterial Proteins/*analysis/immunology/metabolism
;
Blotting, Western
;
Child
;
Child, Preschool
;
Chronic Disease
;
Enzyme-Linked Immunosorbent Assay
;
Female
;
Gastritis/pathology
;
Helicobacter Infections/blood/microbiology/*pathology
;
Helicobacter pylori/isolation & purification/*metabolism
;
Humans
;
Immunoglobulin A/*blood
;
Immunoglobulin G/*blood
;
Infant
;
Infant, Newborn
;
Male
;
Severity of Illness Index
;
Urease/metabolism
2.Proteomics Analysis for Helicobacter pylori-infected Gastric Mucosa.
Ho Suk KANG ; Sung Noh HONG ; Hye Rim PARK ; Mi Jung KWON ; Jun Haeng LEE ; Jae J KIM
The Korean Journal of Gastroenterology 2014;64(1):10-17
BACKGROUND/AIMS: Helicobacter pylori infection is linked to the development of gastric cancer. H. pylori-associated gastric inflammation is considered to be the first important step in the histogenesis of such neoplasia. However, studies that compare proteome of gastric mucosa infected with or without H. pylori are lacking. METHODS: We employed proteomics analysis on the endoscopic biopsy specimens of gastric mucosa obtained from two groups (30 cases): healthy subjects without H. pylori infection (15 cases), and gastritis patients with H. pylori infection (15 cases). The pooled proteins obtained from gastric mucosa infected with or without H. pylori were separated by two-dimensional gel electrophoresis and analyzed by a computer-aided program. The altered protein expressions were then identified by mass spectrometry and validated by Western blotting and immunohistochemistry. RESULTS: On mass spectrometry using MALDI TOF(TM) Analyzer, the up-regulation of Keratin 1, ezrin, adenosine triphosphate (ATP) synthase subunit alpha mitochondrial isoform c, Keratin type I cytoskeletal 19, and Keratin type I cytoskeletal 9 were identified; in contrast, 71 kd heat shock cognate protein, ATP synthase subunit alpha mitochondrial precursor, and annexin IV were down-regulated. Among them, membrane cytoskeleton linker ezrin was validated using Western blot and immunohistochemistry. CONCLUSIONS: Expression of ezrin was significantly different between the gastric mucosa with and without H. pylori infection. Therefore, ezrin could be considered a promising potential molecular marker for detecting H. pylori infection in gastric mucosa.
Blotting, Western
;
Cytoskeletal Proteins/metabolism
;
Down-Regulation
;
Electrophoresis, Gel, Two-Dimensional
;
Female
;
Gastric Mucosa/*metabolism/microbiology
;
Gastritis/complications/metabolism/pathology
;
Gastroscopy
;
Helicobacter Infections/complications/metabolism/*pathology
;
*Helicobacter pylori
;
Humans
;
Immunohistochemistry
;
Male
;
Proteome/*analysis
;
*Proteomics
;
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
;
Up-Regulation
3.Establishment of Mongolian gerbil model of gastric cancer induced by Helicobacter pylori infection and its proteomics analysis.
Yan ZHAO ; Yuan XIE ; Xian CHEN ; Wenjie XU ; Yan WANG ; Jianjiang ZHOU
Chinese Journal of Pathology 2014;43(12):820-826
OBJECTIVETo establish an animal model of gastric cancer by long-term infection of Helicobacter pylori (H.pylori) and to elucidate the pathogenesis by proteomics analysis.
METHODSFifty male Mongolian gerbils (4-5 week-old and weighted 60-100 g) were infected with H.pylori and the gastric tissues were obtained after the infection at 3, 6, 12 and 24 months. Histological changes were evaluated by H-E staining of the gastric tissue sections. Detection of H.pylori was performed by in-vitro culture of fresh gastric tissue samples, PCR amplification of H.pylori 16s rRNA and localization by silver staining. In addition, proteins extracted from gastric tissue samples were subjected to two-dimensional electrophoresis (2-DE) at various infection time points. Protein spots with increased quantity over the course of H.pylori infection were selected and analyzed by LC-MS/MS. Finally, differentially expressed proteins between human gastric cancer tissue samples and lymph nodes were analyzed by real-time RT-PCR.
RESULTSColonization of H.pylori was observed in gastric tissue of gerbils as early as 3 months after H.pylori infection, and persisted till 24 months. Pathological examination of infected animals showed various histological changes including acute gastritis, atrophic gastritis, intestinal metaplasia and gastric carcinoma. Seventy-eight differentially expressed proteins were identified by proteomics analysis, among which 36 proteins were up-regulated and 42 were down-regulated. Analyzed by LC-MS/MS, ten proteins were identified, including lactate dehydrogenase, ATP synthase, fatty acid-binding protein, COX5B, peroxiredoxin-4, peroxide reductase, transgelin, succinyl-CoA ligase, keratin and protein disulfide-isomerase A2, among which transgelin, ATP synthase and lactate dehydrogenase were highly expressed in human gastric carcinoma and lymph nodes.
CONCLUSIONSH.pylori infection induces the expression of transgelin, ATP synthase and lactate dehydrogenase, implying possible roles in the pathogenesis of gastric diseases including cancer.
Animals ; Disease Models, Animal ; Gastritis ; microbiology ; pathology ; Gerbillinae ; Helicobacter Infections ; complications ; metabolism ; Helicobacter pylori ; genetics ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Male ; Metaplasia ; Microfilament Proteins ; metabolism ; Muscle Proteins ; metabolism ; Proteomics ; Proton-Translocating ATPases ; metabolism ; RNA, Ribosomal, 16S ; analysis ; Stomach Neoplasms ; metabolism ; microbiology ; Tandem Mass Spectrometry
4.Effect of huazhuo jiedu hewei recipe on the mechanism of angiogenesis in precancerous lesions of gastric cancer rats.
Shao-fang GAO ; Yan-gang WANG ; Dian-gui LI ; Lin PEI
Chinese Journal of Integrated Traditional and Western Medicine 2013;33(11):1515-1519
OBJECTIVETo explore the possible angiogenesis mechanism of Huazhuo Jiedu Hewei Recipe (HJHR) in preventing and treating precancerous lesions of gastric cancer (PLGC).
METHODSTotally 66 Wistar rats were randomly divided into 6 groups, i.e., the normal control group, the model group, the retinoic acid (RA) group, the high dose HJHR group, the middle dose HJHR group, the low dose HJHR group, 11 in each group. PLGC model was duplicated by inserting a spring with Helicobacter. Corresponding medicines were administered to rats in each medicated group once daily by gastrogavage, 2 mL each time for 12 successive weeks. The effect of HJHR on hypoxia induced factor (HIF-1alpha) and vascular endothelial growth factor (VEGF) of PLGC in chronic atrophic gastritis (CAG) rats' gastric mucosa was observed by immunohistochemical assay and Western blot method.
RESULTSCompared with the normal control group, the expression of VEGF and HIF-1alpha increased in the model group (P < 0.05). Compared with the model group, the expression of VEGF and HIF-1alpha decreased in each medicated group (P < 0.05). Besides, they were lower in the high and middle dose HJHR groups than in the RA group and the low dose HJHR group (P < 0. 05). There was no statistical difference between the low dose HJHR group and the RA group (P > 0.05).
CONCLUSIONHJHR could prevent and treat PLGC of CAG rats possibly through decreasing the expression of HIF-1alpha and VEGF in a dose-dependent manner.
Animals ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis ; metabolism ; microbiology ; Helicobacter ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Neovascularization, Pathologic ; Precancerous Conditions ; blood supply ; drug therapy ; metabolism ; Rats ; Rats, Wistar ; Stomach Neoplasms ; blood supply ; drug therapy ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism
5.Expressions of HSP 70 and NF-kappaB in the peripheral blood lymphocyte of chronic gastritis patients of different syndrome patterns.
Ling HU ; Xiao-Feng ZHENG ; Xue-Hui YAN
Chinese Journal of Integrated Traditional and Western Medicine 2012;32(9):1188-1191
OBJECTIVETo study the expressions of heat shock protein 70 (HSP 70) and nuclear factor-kappa B (NF-kappaB) in the peripheral blood lymphocyte of chronic gastritis (CG) patients of Pi-Wei hygropyrexia syndrome (PWHS) and Pi-qi deficiency syndrome (PQDS), and to explore their correlation with Helicobacter pylori (Hp) infection.
METHODSRecruited were totally 86 CG patients who visited at the clinics of gastroenterology, First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, including 67 patients of PWHS (30 of predominant-dampness, 30 of equal dampness and heat, and 30 of predominant-heat) and 19 patients of PQDS. Another 12 volunteers from healthy employees and students of Guangzhou University of Traditional Chinese Medicine were recruited as the control group. Their peripheral blood was collected. The Hp infection was detected using ASSURE Hp rapid test. The expressions of HSP 70 and NF-kappaB in the peripheral blood lymphocyte were detected using flow cytometry.
RESULTSThe Hp infection rate was 37. 31% in the GS patients of PWHS and 36. 84% in the GS patients of PQDS (P>0.05). Compared with the control group, the expression of HSP 70 decreased in the PWHS predominant-heat group, and the expression of NF-kappaB increased in the PWHS predominant-heat group and the PQDS group (P<0.05). The expression of NF-kappaB were higher in the positive Hp infection patients of PWHS and PQDS than in the control group (P<0.05). The expression of HSP 70 was higher in the positive Hp infection patients of PQDS than in the negative Hp infection patients of PQDS (P<0.05). Besides, the coefficient correlation was -0. 023 between HSP 70 and Hp infection, and 0. 027 between NF-KB and Hp infection (P>0.05).
CONCLUSIONSThe increased expression of NF-KB in the peripheral blood lymphocyte of CG patients of PWHS and PQDS might reflect the pathogenic roles of "inner evil" in Chinese medicine theories. The increased expression of HSP 70 in CG patients of PQDS and decreased expression of HSP 70 in CG patients of PWHS might reflect "vital qi fighting against evils" and "exuberance evils and feeble vital qi" in the body. Hp infection might not be the only factor resulting in the occurrence of PWHS or PQDS.
Adult ; Case-Control Studies ; Female ; Gastritis ; diagnosis ; metabolism ; microbiology ; Gastritis, Atrophic ; diagnosis ; metabolism ; microbiology ; HSP70 Heat-Shock Proteins ; metabolism ; Helicobacter Infections ; diagnosis ; metabolism ; Humans ; Lymphocytes ; metabolism ; Male ; Medicine, Chinese Traditional ; NF-kappa B ; metabolism ; Young Adult
6.The Usefulness of the Regular Arrangement of Collecting Venules Pattern for the Determination of Helicobacter pylori Infection.
Sunyoung NA ; Jun Won CHUNG ; Hyun Joo PARK ; Yoon Jae KIM ; Kwang An KWON ; Ki Baik HAHM ; Duck Joo CHOI ; Seok Hoo JEONG ; Minsu HA ; Geum Ha KIM
The Korean Journal of Gastroenterology 2011;58(5):252-257
BACKGROUND/AIMS: In the Helicobacter pylori (H. Pylori)-negative normal stomach, collecting venules are visible over all the gastric body as numerous minute points evaluated with standard endoscopy. This finding was termed regular arrangement of collecting venules (RAC), and its absence suggests H. Pylori gastritis. The aim of this study was to evaluate the correlation between the RAC and rapid urease test. METHODS: Two hundred sixty three consecutive adults undergoing upper digestive endoscopy and rapid urease test were included. The lesser curvature of the lower corpus was evaluated for the RAC pattern using a standard endoscope and different hemoglobin index. Two biopsies from the lesser curvature of the antrum and the greater curvature of the body were collected for rapid urease test. RESULTS: H. Pylori were detected in 51.3% (135/263) patients. Of the 57 patients with H. Pylori-negative normal stomachs 53 patients (93%) had RAC. As a determinant of the normal stomach without H. Pylori infection, the presence of RAC had 41.4% sensitivity, 97.0% specificity, 93.0% positive predictive value and 63.6% negative predictive value. CONCLUSIONS: RAC-positive finding by standard endoscopy showed high positive predictive value and specificity of H. Pylori-negative normal stomach. RAC-positive finding by standard endoscopy could be an useful finding to predict H. Pylori negativity.
Adult
;
Aged
;
Diagnosis, Differential
;
Endoscopy, Gastrointestinal
;
Female
;
Gastritis/microbiology/pathology
;
Gastroscopy
;
Helicobacter Infections/*diagnosis/microbiology
;
*Helicobacter pylori
;
Hemoglobins
;
Humans
;
Male
;
Middle Aged
;
Pyloric Antrum/blood supply/microbiology/pathology
;
Retrospective Studies
;
Sensitivity and Specificity
;
Urease/metabolism
;
Venules/anatomy & histology
7.Relationship between Pi-Wei damp-heat syndrome with expressions of nuclear factor-kappaB mRNA and heat shock protein 70 mRNA in patients with chronic gastritis.
Na-juan CUI ; Ling HU ; Shao-xian LAO
Chinese Journal of Integrated Traditional and Western Medicine 2010;30(1):18-21
OBJECTIVETo explore the relationship between the genesis of Pi-Wei damp-heat syndrome (PDS) and the Helicobacter pylori (HP) infection in patients with chronic gastritis (CG) by observing the levels of nuclear factor kappaB (NF-kappaB) mRNA and heat shock protein 70 (HSP70) expressions.
METHODSGastric mucous membrane tissues collected through gastroscopy from gastric antrum of 51 CG patients, 36 of PDS type (CG-PDS) and 15 of Pi-qi deficiency syndrome (CG-non-PDS) type, and 8 healthy persons (as control) were examined to diagnose the inflammation level with HE stain and to detect the existence of HP infection by rapid urease test and methylthioninium chloride stain. The mRNA expressions of NF-kappaB and HSP70 in the tissues were determined quantitatively with FQ-PCR as well.
RESULTSPatients of CG-PDS showed higher level of HSP70 mRNA expression than in the control; and level of NF-kappaB mRNA expression was higher than that in the control and CG-non-PDS (all P<0.05); but both expressions were insignificantly different in CG-PDS patients with positive or negative HP infection (P>0.05). Positive correlation of the two expressions was shown in CG-PDS with negative HP infection.
CONCLUSIONNF-kappaB and HSP70 may partially embody, in some extent, the power of vital qi and evil qi in the organism; the over-expressed NF-kappaB and HSP70 may indicate the severe fighting between evil qi and vital qi, and both would be influenced to a certain degree in the fighting process.
Adult ; Case-Control Studies ; Female ; Gastric Mucosa ; metabolism ; Gastritis ; diagnosis ; metabolism ; microbiology ; HSP70 Heat-Shock Proteins ; genetics ; metabolism ; Helicobacter Infections ; metabolism ; Helicobacter pylori ; Humans ; Male ; Medicine, Chinese Traditional ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; genetics
8.Expressions of connexin 32 and connexin 43 in patients with gastric precancerous lesion after eradication of Helicobacter pylori.
Yan JIA ; Can-Xia XU ; Wen-Bin YANG
Journal of Central South University(Medical Sciences) 2008;33(7):628-633
OBJECTIVE:
To observe the change in expressions of connexin 32 and connexin 43 after the eradication of Helicobacter pylori (H.pylori) in patients with gastric precancerous lesion.
METHODS:
The expressions of connexin 32 and connexin 43 in gastric mucosa specimens biopsy under endoscopy were detected by immunohistochemistry. The expressions of connexin 32 and connexin 43 were detected before and after the eradication of H.pylori in 88 patients with gastric precancerous lesion, and in 33 patients with chronic superficial gastritis (CSG).
RESULTS:
The positive expression rates and the expressional intensity of connexin 32 and connexin 43 in patients with gastric precancerous lesions (51.1% and 54.5%) were lower than those in patients with CSG (100% and 93.9%, P < 0.05).In patients with gastric precancerous lesions,the positive expression rates and the expressional intensity of connexin 32 and connexin 43 in H.pylori positive group (41.4% and 44.8%) were lower than those in H.pylori negative group (70% and 73.3%, P < 0.05). In gastric precancerous lesions group, the positive expression rates of connexin 32 and connexin 43 in H.pylori positive group before the eradication therapy (41.4% and 44.8%, respectively) was lower than those after the eradication of H.pylori (97.9% and 91.7%, P < 0.05); in the eradication failure group, the positive expression rates of connexin 32 and connexin 43 were 40% and 50%. The eradication failure group before the treatment and after the treatment had no statistical significance(P > 0.05).
CONCLUSION
The expressions of connexin 32 and connexin 43 in patients with gastric precancerous lesions are low, and the eradication of H.pylori can upregulate their expressions.
Adult
;
Aged
;
Connexin 43
;
biosynthesis
;
Connexins
;
biosynthesis
;
Female
;
Gastritis
;
metabolism
;
microbiology
;
Helicobacter Infections
;
drug therapy
;
metabolism
;
Helicobacter pylori
;
Humans
;
Male
;
Middle Aged
;
Precancerous Conditions
;
metabolism
;
microbiology
;
Stomach Neoplasms
;
metabolism
;
microbiology
9.The Role of Gastric Acid in the H. pylori-induced Gastritis in Mouse.
Sung Soo KIM ; Yeo Min LEE ; Hyung Keun KIM ; Jin Il KIM ; Hiun Suk CHAE ; In Sik CHUNG ; Doo Ho PARK
The Korean Journal of Gastroenterology 2007;50(6):363-369
BACKGROUND/AIMS: This study was designed to investigate the role of gastric acid in the extent of H. pylori-induced gastritis. METHODS: Twenty eight mice were innoculated with live H. pylori. They were allocated into four groups. Mice in group I received no treatment, group II mice were treated with sham injection, group III received 125microgram/kg body weight of pentagastrin, while group IV received 250microgram/kg body weight of pentagastrin subcutaneously three times a week. After 7 months, the mucosal pH, H. pylori density, neutrophils and monocytes infiltration, and the degree of atrophy were assessed in the stomach. RESULTS: In the gastric body, the densities of H. pylori were not different among groups. The degree of neutrophil infiltration was significantly lower in group IV compared to other groups (p<0.05). The degree of monocyte infiltration was also significantly lower in group IV than group III (p<0.05). In the gastric antrum, there was no significant difference of the H. pylori density, neutrophil and monocyte infiltration, and degree of atrophy among the groups. The mice with the gastric mucosal pH lower than mean of 3.2 had significant lower level of H. pylori density (1.4 vs. 2.4, p=0.04), and infiltration of neutrophils (0.9 vs. 2.3, p=0.018), and monocytes (1.2 vs. 1.8; p=0.011) than the those with mucosal pH above 3.2 in the body of stomach. CONCLUSIONS: Gastric acid plays a role in suppressing the proximal propagation of H. pylori-induced gastritis to the body of stomach.
Animals
;
Female
;
Gastric Acid/*metabolism
;
Gastric Mucosa/pathology
;
Gastritis/immunology/*microbiology
;
Helicobacter Infections/*immunology/microbiology
;
*Helicobacter pylori/isolation & purification
;
Hydrogen-Ion Concentration
;
Mice
;
Mice, Inbred C57BL
;
Models, Animal

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