Isolation and diagnosis of Helicobacter pylori by a new method: Microcapillary culture

AIM:To investigate the performance of the microcapillary culture method(MCM) in Helicobacter pylori(H.pylori) isolation and diagnosis.METHODS:Microcapillary culture(MC),classical culture(CC),rapid urease(CLO) test,and histopathologic examination(HE) were performed with biopsy samples.Homogenized biopsy samples were loaded into capillary tubes and incubated for 48 h at 37 ℃ without providing a microaerophilic environment.Additionally,three or four loops of the homogenized sample were inoculated in a ready-to-use selective medium(Becton Dickinson,Helicobacter Agar,Modified) specific for the isolation of H.pylori and incubated at 37 ℃ in a microaerophilic atmosphere provided by Campy Gen(Becton Dickinson,Gas Pack).Bacteria reproducing in microcapillary tubes were evaluated in an inverted microscope and also were evaluated after performing a CC with the content.Results obtained by CC,CLO test,and HE were compared with those of MC.The diagnostic performances of the methods used in this study were evaluated for specificity,sensitivity,positive predictive value(PPV),negative predictive value(NPV),and CI.RESULTS:H.pylori was found positive by CLO test +HE and/or CC culture in 26 patient antrum and corpus biopsy samples.In 25(25/26) patient biopsy samples,H.pylori was isolated by MCM,whereas in only 14(14/26) patient biopsy samples,H.pylori was isolatedby CC.CLO test and HE were found positive in 17(17/26) patient biopsy samples.Comparing the results of the isolation of H.pylori by MCM,CC,CLO test,and HE,the sensitivity of the MCM was found as 96%,the specificity as 80%,the PPV as 83%,the NPV as 95%,and the 95%CI as 0.76(χ2 =31.51,P < 0.01) whereas the sensitivity of the CC was found as 54%(χ2 =19.15,P < 0.01),and the sensitivity of the CLO test and HE were found as 65%(χ2 =25.26,P < 0.01).CONCLUSION:This new microcapillary cultivation method for H.pylori has high diagnostic sensitivity compared with CC,HE,and CLO tests.     

Detection of Helicobacter pylori in Gastric Aspirates Using a Monoclonal Antibody-Based Test

Background/Aims

The objective of this study was to evaluate a monoclonal antibody-based test to detect Helicobacter pylori-specific antigen in gastric aspirates from humans.

Methods

Sixty-one volunteers were enrolled in the study. All of the subjects underwent a ¹³C-urea breath test (UBT) before esophagogastroduodenoscopy. Gastric aspirates were analyzed for pH and ammonia and used for polymerase chain reaction (PCR), culture, and monoclonal antibody-based detection of H. pylori. Multiple biopsies of the gastric antrum and body were obtained for a rapid urease test (RUT) and histological evaluation.

Results

Thirty-six subjects were H. pylori-positive and 25 were H. pylori-negative according to the UBT results. Compared with the H. pylori-negative subjects, H. pylori-positive subjects had a higher pH (4.77±1.77 vs 3.49±1.30, p<0.05) and ammonia level (1,130.9±767.4 vs 184.2±126.3, p<0.0001). The sensitivities and specificities of the PCR test, RUT, culture test, and monoclonal antibody-based test were 100% and 72%, 89% and 100%, 47% and 100%, and 78% and 100%, respectively.

Conclusions

The monoclonal antibody-based test for diagnosing H. pylori infection in gastric aspirates has increased sensitivity compared with the culture test and specificity as high as that of the RUT. The test may be useful as an additive test for examining gastric aspirates.     

Seroepidemiology of Helicobacter pylori infection in elderly people in the Beijing region,China

AIM:To investigate seroepidemiology of cagA+and vacA+strains of Helicobacter pylori(H.pylori)in an elderly population in Beijing and to determine risk factors for seropositivity.METHODS:A total of 2006 elderly persons(>60years)were selected using a random cluster sampling method in different parts of the Beijing area(urban,suburban and mountainous districts).Structured questionnaires were completed during home visits,including history of H.pylori infection,history of gastrointestinal diseases,diet types,hygiene habits,occupation and economic status.Blood samples(2 mL)were collected from each participant,and serum IgG antibodies to cagA,vacA and H.pylori urease antigens were measured by immunodetection.RESULTS:The prevalence of H.pylori infection in elderly subjects was 83.4%and the typeⅠH.pylori strain infection rate was 56%.The seroprevalence for typeⅠH.pylori strain infection in urban and suburbandistricts was higher than that in the mountainous areas(P<0.001).Elderly subjects who had previously performed manual labor or were in the young-old age group(age<75 years)had a higher seroprevalence of H.pylori infection than those who had previously performed mental labor or were in the oldest-old age group(age≥75 year)(P<0.05).The typeⅠH.pylori strain infection rate in the elderly with vegetarian diets was higher than in those eating high-protein foods(P<0.001).There was no significant difference in the prevalence of H.pylori strains between male and female elderly participants(P>0.05).CONCLUSION:TypeⅠH.pylori seroprevalence is higher in elderly people.The distribution of strains of H.pylori is significantly affected by age,area and dietary habits.     

Simple animal model of Helicobacter pylori infection

Helicobacter pylori(H.pylori)has become accepted as a human pathogen for the development of gastritis and gastroduodenal ulcer.To develop a simple rat model of chronic H.pylori infection,male Sprague-Dawley rats were pretreated with streptomycin suspended in tap water(5 mg/mL)for 3 d.The rats were inoculated by gavage at 1 mL/rat with H.pylori suspension(5×108-5×1010 CFU/mL)twice daily at an interval of 4 h for three consecutive days.Two weeks after inoculation,rats were sacrificed and the stomachs were removed.Antral biopsies were performed for urease test and the stomachs were taken for histopathology.Successful H.pylori inoculation was defined as a positive urease test and histopathology.We reported a 69.8%-83.0%success rate for H.pylori infection using the urease test,and hematoxylin and eosin staining confirmed the results.Histopathological analysis detected bacteria along the mucous lining of the surface epithelium and crypt lumen and demonstrated mild to moderate gastric inflammation in successfully inoculated rats.We developed a simple rat model of chronic H.pylori infection for research into gastric microcirculatory changes and therapy with plant products.     

Value of a new stick-type rapid urine test for the diagnosis of Helicobacter pylori infection in the Vietnamese population

AIM:To assess the value of a new test for the diagnosis of Helicobacter pylori(H.pylori) infection,Rapirun H.pylori Antibody Stick(Rapirun Stick),in a Vietnamese population.METHODS:Eligible patients without previous history of H.pylori eradication were recruited.Rapid urease test(RUT) and histologic examination were used to diagnose the H.pylori infection.Patients were considered H.pylori positive when the RUT results were positive and/or the bacteria were detected histologically.Rapirun Stick tests were performed using urine samples,and the results were compared with the other 2 methods.RESULTS:We enrolled 200 patients with a mean age of 36(range,18-76) years.There were 116 females and 84 males.Of the 200 patients,111(55.5%) were diagnosed as being H.pylori positive.The sensitivity,specificity,and accuracy of the Stick test were 84.7%,89.9%,and 87.0%,respectively.There were 17(8.5%) falsenegative patients and 9(4.5%) false-positive patients.CONCLUSION:The Rapirun Stick test has high sensitivity,specificity,and accuracy for the diagnosis of H.pylori infection in the Vietnamese population.The test can be clinically applied in Vietnamese populations.     

Diagnostic performance of gastric imprint smear for determination of Helicobacter pylori infection

BACKGROUND:

Despite the availability of several methods (invasive and noninvasive) for the diagnosis of Helicobacter pylori infection, no test is considered to be the ‘gold standard’. Endoscopy-based tests are regarded as the reference method in most studies.

OBJECTIVE:

To evaluate the diagnostic performance of imprint cytology smears of antral biopsies compared with Gram-stained smears, the rapid urease test and culture methods, separately and in combination.

METHODS:

Antral biopsies were obtained from consecutive patients undergoing upper gastrointestinal endoscopy at a single centre. The biopsies were examined for the presence of H pylori by Gram-stained smear, the rapid urease test, culture methods and imprint cytology smear.

RESULTS:

A total of 273 biopsies were studied. All tests were positive in 36% of the patients. Of 252 biopsies tested, 73% were positive using the imprint cytology technique. Using Gram-stained smear, the rapid urease test and culture methods individually, the sensitivity and specificity of imprint cytology smears for the detection of H pylori were found to be 92.7% and 50%; 92.7% and 49%; and 92.4% and 38.5%, respectively. Combining the three microbiological methods resulted in a sensitivity of 92.1%, a specificity of 51.0% and an efficiency of 71.7% for imprint cytology smears.

CONCLUSIONS:

Endoscopic examination provides useful clinical information. Imprint gastric cytology can be used as a rapid test to establish the diagnosis of H pylori infection at the time endoscopy is performed, enabling the endoscopist to start treatment with immediate effect.     

Different risk factors influence peptic ulcer disease development in a Brazilian population

AIM: To investigate age, sex, histopathology and Helicobacter pylori (H. pylori) status, as risk factors for gastroduodenal disease outcome in Brazilian dyspeptic patients.tients submitted to upper gastroscopy at Hospital das Clinicas of Marilia, antral biopsy specimens were obtained and subjected to histopathology and H. pylori diagnosis. All patients presenting chronic gastritis (CG) and peptic ulcer (PU) disease localized in the stomach, gastric ulcer (GU) and/or duodenal ulcer (DU) were included in the study. Gastric biopsies (n = 668) positive for H. pylori by rapid urease test were investigated for vacuolating cytotoxin A (vacA ) medium (m) region mosaicism by polymerase chain reaction. Logistic regression analysis was performed to verify the association of age, sex, histopathologic alterations, H. pylori diagnosis and vacA m region mosaicism with the incidence of DU, GU and CG in patients. RESULTS: Of 1466 patients submitted to endoscopy, 1060 (72.3%) presented CG [male/female = 506/554; mean age (year) ± SD = 51.2 ± 17.81], 88 (6.0%) presented DU [male/female = 54/34; mean age (year) ± SD = 51.4 ± 17.14], and 75 (5.1%) presented GU [male/female = 54/21; mean age (year) ± SD = 51.3 ± 17.12] and were included in the comparative analysis. Sex and age showed no detectable effect on CG incidence (overall c 2 = 2.1, P = 0.3423). Sex [Odds ratios (OR) = 1.8631, P = 0.0058] but not age (OR = 0.9929, P = 0.2699) was associated with DU and both parameters had a highly significant effect on GU (overall c 2 = 30.5, P 0.0001). The histopathological results showed a significant contribution of ageing for both atrophy (OR = 1.0297, P 0.0001) and intestinal metaplasia (OR = 1.0520, P 0.0001). Presence of H. pylori was significantly associated with decreasing age (OR = 0.9827, P 0.0001) and with the incidence of DU (OR = 3.6077, P 0.0001). The prevalence of m1 in DU was statistically significant (OR = 2.3563, P = 0.0018) but not in CG (OR = 2.678, P = 0.0863) and GU (OR = 1.520, P = 0.2863). CONCLUSION: In our population, male gender was a risk factor for PU; ageing for GU, atrophy and metapla-sia; and H. pylori of vacA m1 genotype for DU.     

Significant decrease in prevalence of Helicobacter pylori in the Czech Republic

AIM: To study possible decrease in prevalence of Helicobacter pylori (H. pylori) infection in the Czech Republic within a 10-year period.METHODS: A total of 22 centres entered the study. The catchment areas of these centres covered cities and towns with more than 20 000 inhabitants, smaller towns (≤ 20 000 inhabitants) with surrounding villages and rural areas, and were spread over the whole country, corresponding well to the geographical distribution of the Czech population. A total of 1 837 subjects (aged 5-98 years) took part in the study, randomly selected out of 38 147 people from the general population. H. pylori infection was investigated by means of a ¹³C-urea breath test. Breath samples in duplicates were analysed using isotope ratio mass spectrometry. The cut-off point was 3.5. Social and demographic characteristics were based on data from self-completed questionnaires.RESULTS: The overall prevalence of H. pylori infection was 23.5% (430/1826), and 4.8% (20/420) in children aged 15 or less. There was no statistically significant difference in prevalence between males (24.3%; 208/857) and females (22.9%, 222/969, P = 0.494). H. pylori infection was strongly associated with higher age, among subjects aged 55+ years, prevalence of H. pylori infection was 39.8% (252/633, P < 0.001). The highest prevalence of H. pylori infection was found among persons aged 55-64 years (43.9%, 97/221) and 75+ years (37.9%, 58/153). Among study subjects aged 15+ years, prevalence of H. pylori infection was significantly increased in those with lowest education (odds risk 3.19, 95% CI 1.87-5.47). Compared to never married (14.1%), the prevalence of H. pylori infection was statistically significantly higher among married (35.4%, 246/694, P < 0.001), divorced (36.8%, 49/133, P < 0.001) and widowed study subjects (40.2%, 45/112, P < 0.001), both in minimally and fully adjusted analysis. There was no significant difference in the prevalence of H. pylori infection between married and widowed subjects (35.4%, 246/694 vs 40.2%, 45/112, P = 0.389). There was little variation in smoking prevalence across categories of smoking and there was no evidence of an increased risk of H. pylori infection among current or past smokers in our data (odds risk 1.04 with 95% CI 0.78-1.40 for current smokers; odds ratio 0.83 with 95% CI 0.60-1.16 for former smokers). The current prevalence of H. pylori in 2011 was significantly lower compared to the prevalence reported from identical geographical areas in 2001 (23.5% vs 41.7%, P < 0.001).CONCLUSION: The overall prevalence of H. pylori infection in the general population has fallen substantially in the Czech Republic over the past 10 years.     

Hypermethylation of TGF-β1 gene promoter in gastric cancer

AIM:To examine transforming growth factor-β1(TGF-β1)promoter methylation in gastric cancer and to determine if Helicobacter pylori(H.pylori)or interleukin(IL)-1β could induce TGF-β1 hypermethylation in vitro.METHODS:We examined the frequency and extent of TGF-β1 promoter methylation using methylationspecific PCR in the gastric tissues from 47 gastric cancer patients and 39 non-gastric cancer subjects.H.pylori infection was confirmed by a positive result from either a serological test,histological analysis or C13urea breath test.GES-1 and MKN-45 cells co-cultured with H.pylori or treated with IL-1β for 12,24 and 48 h in vitro tested the effects of H.pylori or IL-1β on TGF-1β.RESULTS:Twenty-four/forty-seven(51%)cases of gastric cancer(GC)tissues showed TGF-β1 promoter methylation,15/47(31.9%)cases of matched noncancerous gastric mucosa tissues from the GC patients,and 11/39(28%)case of the normal gastric mucosa tissues from non-GC subjects showed TGF-β1 promoter methylation(51%vs 28%,P<0.05).Significantly higher levels of methylation of TGF-β1 were found in the tumor tissues than in non-tumor tissues from GC patients(0.24±0.06 vs 0.17±0.04,P<0.05)and normal gastric tissues from non-GC subjects(0.24±0.06 vs 0.15±0.03,P<0.05).TGF-β1 methylation was found in 48.3% of H.pylori-positive gastric mucosal tissues whereas only 23.1% of H.pylori-negative gastric mucosal tissues showed TGF-β1 methylation(48.3%vs 23.1%,P<0.05).IL-1β appeared to induce a dose-dependent methylation of TGF-β1 and the strongest methylation was observed in GES-1 cells treated with 2.5 ng/mL of IL-1β for 48 h.Further studies showed that pre-treatment of GES-1 cells with 20ng/mL IL-1RA for 1 h could partially abolish the effect of IL-1β on TGF-β1 methylation.Infection of GES-1cells by H.pylori was not found to induce significant TGF-β1 promoter methylation.CONCLUSION:Our data revealed that TGF-1 promoter is methylated in GC patients.IL-1β may be an important mediator for H.pylori induced gene methylation during GC     

Serum pepsinogen Ⅱ is a better diagnostic marker in gastric cancer

AIM:To investigate screening makers for gastric cancer,we assessed the association between gastric cancer and serum pepsinogens(PGs).METHODS:The subjects comprised 450 patients with gastric cancer,111 individuals with gastric atrophy,and 961 healthy controls.Serum anti-Helicobacter pylori(H.pylori) immunoglobulin G(IgG),PGⅠand PG Ⅱ were detected by enzyme-linked immunosorbent assay.Gastric atrophy and gastric cancer were diagnosed by endoscopy and histopathological examinations.Odds ratios and 95%CIs were calculated using multivariate logistic regression.RESULTS:Rates of H.pylori infection remained high in Northeastern China.Rates of H.pylori IgG positivity were greater in the gastric cancer and gastric atrophy groups compared to the control group(69.1% and 75.7% vs 49.7%,P 0.001).Higher levels of PG Ⅱ(15.9 μg/L and 13.9 μg/L vs 11.5 μg/L,P 0.001) and lower PGⅠ/PG Ⅱ ratio(5.4 and 4.6 vs 8.4,P 0.001) were found in patients with gastric cancer or gastric atrophy compared to healthy controls,whereas no correlation was found between the plasma PGⅠconcentration and risk of gastric cancer(P = 0.537).In addition,multivariate logistic analysis indicated that H.pylori infection and atrophic gastritis were independent risk factors for gastric cancer.Lower plasma PGⅠ/PG Ⅱ ratio was associated with higher risks of atrophy and gastric cancer.Furthermore,plasma PG Ⅱ?level?significantly?correlated?with?H.pyloriinfected gastric cancer.CONCLUSION:Serum PG Ⅱ concentration and PG Ⅰ/PG Ⅱ ratio are potential biomarkers for H.pyloriinfected gastric disease.PG Ⅱ is independently associated with risk of gastric cancer.     

Helicobacter pylori acquistion of metronidazole resistance by natural transformation in vitro

AIM: To study whether Helicobacter pylori is naturally transformable.METHODS: Transformation was performed in BHI broth supplemented with horse serum and yeast extract. Genomic DNA extracted from a metronidazole resistant H.pylori strain was added to H. pylori broth culture. The mixture was incubated at microaerophilic atmosphere. The DNA-treated cells were plated on blood agar containing 8 mg/L metronidazole to select for transformants. Sterile distilled water was used as a negative DNA control. The DNA profiles of transformants were compared with that of their parent strains by randomly amplified polymorphic DNA (RAPD) fingerprinting.RESULTS: Transformation of H. pylori with DNA from a metronidazole resistant strain as a marker was demonstrated. Out of the 12 strains of H. pylori tested, 9 (75%) strains were found to be transformable. The transformation frequencies ranged from 3.4 × 10^-6 to 2.4 × 10^-4. By RAPD, DNA fingerprints of the transformants and their parent strains showed no change in DNA profiles though transformants were all resistant to metronidazole as compared with their metronidazole-sensitive parent strains.CONCLUSION: Helicobacter pylori is naturally transformable which might be one of the ways that H. pylori develops resistance to metronidazole.     

Abnormal DNA-PKcs and Ku 70/80 expression may promote malignant pathological processes in gastric carcinoma

AIM:To determine the expression of the catalytic subunit of DNA-dependent protein kinase(DNA-PKcs)and the Ku70/Ku80 heterodimer(Ku 70/80)in gastric carcinoma.METHODS:Gastric biopsies were obtained from 146gastric carcinoma patients[Helicobacter pylori(H.pylori)-negative:89 and H.pylori-positive:57]and 34from normal subjects(H.pylori-negative:16 and H.pylori-positive:18)via surgery and endoscopic detection from April 2011 to August 2012 at the First Affiliated Hospital of Nanchang University.Pathological diagnosis and classification were made according to the criteria of the World Health Organization and the updated Sydney system.An‘‘in-house’’rapid urease test and modified Giemsa staining were employed to detect H.pylori infection.The expression of DNA-PKcs and the Ku 70/80protein was detected by immunohistochemistry.RESULTS:Overall,the positive rates of both DNA-PKcs and Ku 70/80 were significantly increased in gastric cancer(χ2=133.04,P<0.001 for DNA-PKcs andχ2=13.06,P<0.01 for Ku)compared with normal gastric mucosa.There was hardly any detectable expression of DNA-PKcs in normal gastric mucosa,and the positive rate of DNA-PKcs protein expression in patients with a normal gastric mucosa was 0%(0/34),whereas the rate in gastric cancer(GC)was 93.8%(137/146).The difference between the two groups was statistically significant.Additionally,the positive rate of Ku 70/80 was79.4%(27/34)in normal gastric mucosa and 96.6%(141/146)in gastric cancer.The DNA-PKcs protein level was significantly increased in gastric cancer(MannWhitney U=39.00,P<0.001),compared with normal gastric mucosa.In addition,there was a significant difference in the expression of Ku 70/80(Mann-Whitney U=1117.00,P<0.001)between gastric cancer and normal gastric mucosa.There was also a significant difference in Ku70/80 protein expression between GC patients with and without H.pylori infection(P<0.05).Spearman analysis showed a negative correlation between tumor differentiation and DNA-PKcs expression(r=-0.447,P<0.05).Moreov     

High rate of Helicobacter pylori reinfection in Lithuanian peptic ulcer patients

AIM: To evaluate the frequency of Helicobacter pylori(H. pylori) reinfection in peptic ulcer patients during 9 years after H. pylori eradication.METHODS: We invited 117 peptic ulcer patients in whom eradication of H. pylori was confirmed 1 year after eradication treatment both by histology and by rapid urease test. In total, 57 patients were available for the study procedures: 34(59.6%) male, 23(40.4%) female; mean age 52.3 ± 13.0 years. There were 45(78.9%) patients with duodenal ulcer and 12(21.1%) with gastric ulcer. H. pylori was diagnosed by a rapid urease test and histology if endoscopy was performed. If endoscopy was refused, H. pylori was diagnosed by the C14-urea breath test and serology. H. pylori was established if at least one of the tests was positive.RESULTS: The mean follow-up was 8.9 ± 1.0 years(range, 6-12). H. pylori was established in 15 patients. In 2 H. pylori-negative patients, H. pylori was established during the follow-up period and eradicated. Therefore, we consider that reinfection occurred in 17 patients. In the per protocol analysis, reinfection was established in 17 of 57(29.8%; 95%CI: 19.2-42.2) patients during the follow-up period. The annual rate of infection was 3.36%. If all non-responders were considered H. pylori-negative, reinfection would be 14.5%(17/117), the annual ratebeing 1.63%. The mean age of patients with reinfection was 51.8 ± 14.0 years, and without reinfection was 52.5 ± 13.0 years, P 0.05; the mean body mass index of patients with reinfection was 27.2 ± 4.1 kg/m2, and without reinfection was 25.7 ± 4.2 kg/m2, P 0.05. There were no differences in the reinfection rates according the location of the peptic ulcer, the eradication regimen used, and smoking status.CONCLUSION: The reinfection rate of H. pylori is relatively high in Lithuania and probably related to the high prevalence of H. pylori, what may reflect differences in the socioeconomic status between Western and Eastern European countries.     

Helicobacter pylori infection is associated with gallstones: Epidemiological survey in China

AIM: To elucidate the prevalence and risk factors for gallstones, primarily focusing on Helicobacter pylori(H. pylori) infection. METHODS: A total of 10016 Chinese subjects, who had undergone physical examination, fasting 13 C urea breath test and abdominal ultrasonography, had sufficient blood test data, and had finished a questionnaire, were included in this cross-sectional study. Participants(n = 1122) who had previous eradication of H. pylori were studied separately. RESULTS: Gallstones were discovered in 9.10% of men and 8.58% of women, with no significant sex difference. Multivariate analyses displayed that age, aspartate aminotransferase, total cholesterol, H. pylori infection, hepatitis C virus(HCV) infection, and fattyliver had a significant association with gallstones(P 0.05). Successive multiple logistic regression analysis including index of odds ratio(OR) and standardized coefficient(β) indicated that older age(OR/β = 1.056/0.055), H. pylori infection(OR/β = 1.454/0.109), HCV infection(OR/β = 1.871/0.123), and fatty liver(OR/β = 1.947/0.189) had a significant positive association with gallstones. After age stratification, H. pylori infection and fatty liver still had a significant positive association with gallstones in any age-specific groups, whereas HCV infection had a significant positive association in patients aged 40 years. The prevalence of gallstones among H. pylori-positive, H. pylori-eradicated, and H. pylori-negative subjects was 9.47%, 9.02%, and 8.46%, respectively. The matched analysis showed that gallstones among H. pylori eradicated subjects was significantly lower compared with H. pylori-positive subjects(P 0.05).CONCLUSION: H. pylori infection and fatty liver have a significant positive association with gallstones. H. pylori eradication may lead to prevention of gallstones.     

Helicobacter pylori eradication in patients with chronic immune thrombocytopenic purpura

AIM:To assess the effect of Helicobacter pylori(H.pylori)eradication on platelet counts in patients with chronic immune thrombocytopenic purpura(cITP).METHODS:A total of 36 cITP patients were included in the study.The diagnosis of H.pylori was done by rapid urease test and Giemsa staining of the gastric biopsy specimen.All H.pylori positive patients received standard triple therapy for 14 d and were subjected for repeat endoscopy at 6 wk.Patients who continued to be positive for H.pylori on second endoscopyreceived second line salvage therapy.All the patients were assessed for platelet response at 6 wk,3rd and 6th months.RESULTS:Of the 36 patients,17 were positive for H.pylori infection and eradication was achieved in16 patients.The mean baseline platelet count in the eradicated patients was 88615.38±30117.93/mm3and platelet count after eradication at 6 wk,3 mo and6 mo was 143230.77±52437.51/mm3(P=0.003),152562.50±52892.3/mm3(P=0.0001),150187.50±41796.68/mm3(P=0.0001)respectively and in the negative patients,the mean baseline count was71000.00±33216.46/mm3 and at 6 wk,3rd and 6th month follow up was 137631.58±74364.13/mm3(P=0.001),125578.95±71472.1/mm3(P=0.005),77210.53±56892.28/mm3(P=0.684)respectively.CONCLUSION:Eradication of H.pylori leads to increase in platelet counts in patients with cITP and can be recommended as a complementary treatment with conventional therapy.     

Lactobacillus plantarum B7 inhibits Helicobacter pylori growth and attenuates gastric inflammation

AIM:To determine the anti-Helicobacter property of Lactobacillus plantarum B7(L.plantarum)B7 supernatants in vitro and the protective effects of L.plantarum B7 on serum tumor necrosis factor-alpha(TNF-?),gastric malondialdehyde(MDA)level,apoptosis,and histopathology in Helicobacter pylori(H.pylori)-induced gastric inflammation in rats. METHODS:In vitro,the inhibition of H.pylori growth was examined using L.plantarum B7 supernatants at pH 4 and pH 7 and at the concentration of 1×,5×and 10×on plates inoculated with H.pylori.The inhibitory effect of H.pylori was interpreted by the size of the inhibition zone.In vitro,male Sprague-Dawley rats were randomly divided into four groups including group 1(control group),group 2(H.pylori infected group), group 3(H.pylori infected with L.plantarum B7 106 CFUs/mL treated group)and group 4(H.pylori infected with L.plantarum B7 1010 CFUs/mL treated group).One week after H.pylori inoculation,L.plantarum B7 106 CFUs/mL or 10 10 CFUs/mL were fed once daily to group 3 and group 4,respectively,for one week.Blood and gastric samples were collected at the end of the study. RESULTS:In vitro,at intact pH 4,mean inhibitory zone diameters of 8.5 mm and 13 mm were noted at concentrations of 5×and 10×of L.plantarum B7 supernatant disks,respectively.At adjusted pH 7, L.plantarum B7 supernatants at concentrations of 5 ×and 10×yielded mean inhibitory zone diameters of 6.5 mm and 11 mm,respectively.In the in vitro study, in group 2,stomach histopathology revealed mild to moderate H.pylori colonization and inflammation.The level of gastric MDA and epithelial cell apoptosis were significantly increased compared with group 1.The serum TNF-??level was significant decreased in group 3 compared with group 2(P0.05).In addition,L.plantarum B7 treatments resulted in a significant improvement in stomach pathology,and decreased gastric MDA level and apoptotic epithelial cells. CONCLUSION:L.plantarum B7 supernatant inhibits H.pylori growth.This inhibition was dose-dependent and greater at pH 4.Moreover,L.plantarum B7 attenuated H.pylori-induced gastric inflammation.     

First-line eradication for Helicobacter pylori-positive gastritis by esomeprazole-based triple therapy is influenced by CYP2C19 genotype

AIM: To evaluate the effect of first line esomeprazole (EPZ)-based triple therapy on Helicobacter pylori (H. pylori) eradication.METHODS: A total of 80 Japanese patients with gastritis who were diagnosed as positive for H. pylori infection by endoscopic biopsy-based or ¹³C-urea breath tests were included in this study. The average age of the patients was 57.2 years (male/female, 42/38). These patients were treated by first-line eradication therapy with EPZ 40 mg/d, amoxicillin 1500 mg/d, and clarithromycin 400 mg/d for 7 d. All drugs were given twice per day. Correlations between H. pylori eradication, CYP2C19 genotype, and serum pepsinogen (PG) level were analyzed. This study was registered with the UMIN Clinical Trials Registry (UMIN000009642).RESULTS: The H. pylori eradication rates by EPZ-based triple therapy evaluated by intention-to-treat and per protocol were 67.5% and 68.4%, respectively, which were similar to triple therapies with other first-generation proton pump inhibitors (PPIs). The eradication rates in three different CYP2C19 genotypes, described as extensive metabolizer (EM), intermediate metabolizer, and poor metabolizer, were 52.2%, 72.1%, and 84.6%, respectively. The H. pylori eradication rate was significantly lower in EM than non-EM (P < 0.05). The serum PG I level and PG I/II ratio were significantly increased after eradication of H. pylori (P < 0.01), suggesting that gastric atrophy was improved by H. pylori eradication. Thus, first-line eradication by EPZ-based triple therapy for patients with H. pylori-positive gastritis was influenced by CYP2C19 genotype, and the eradication rate was on the same level with other first-generation PPIs in the Japanese population.CONCLUSION: The results from this study suggest that there is no advantage to EPZ-based triple therapy on H. pylori eradication compared to other first-generation PPIs.     

Antibiotic Susceptibility Profile of Helicobacter pylori Isolated from the Dyspepsia Patients in Tehran, Iran

Background/Aim:

Helicobacter pylori is an important pathogen for gastroduodenal diseases. Infection with H. pylori can be limited by regimens of multiple antimicrobial agents. However, antibiotic resistance is a leading cause of treatment failure. The aim of this study has been to determine the resistance patterns of H. pylori strains isolated from gastric biopsies of patients with dyspepsia by agar dilution method, in Tehran, Iran

Patients and Methods:

H. pylori isolates from patients with gastrointestinal diseases were evaluated for susceptibility testing by agar dilution method. Susceptibility testing was performed to commonly used antibiotics including clarithromycin, tetracycline, amoxicillin, metronidazole and ciprofloxacin.

Results:

Among 92 patients with dyspepsia, H. pylori strains were isolated from 42 patients. Seventeen (40.5%) of the isolates were resistant to metronidazole (MICs ≥ 8 μg/l), whereas one isolate (2.4%) was resistant to amoxicillin (MICs ≤ 0. 5 μg/ml) and ciprofloxacin (MICs ≤ 1μg/ml). The resistance rates to other antibiotics in H. pylori isolates are recorded as follows: clarithromycin 6 (14.3 %), tetracycline 2 (4.8%). In 5 of 42 resistant cases, combined resistance was found.

Conclusions:

These data suggest that metronidazole should be used among Iranian patients in first-line therapy with caution, and ciprofloxacin in association with amoxicillin and a proton pump inhibitor is more recommended.