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Objective: The objective of this study is to develop a prediction model for the pathological upgrading of low-grade dysplasia (LGD) in gastric mucosa. The study aims to compare the performance of a traditional model based on clinical and endoscopic factors with an enhanced model that incorporates AMACR staining of biopsy tissues. Methods: The study utilized a training dataset of 405 LGD cases to establish and compare the traditional and enhanced prediction models. Factors associated with upgrading were identified, and the traditional model was based on these factors. The enhanced model incorporated AMACR staining. The models' performances were evaluated using the area under the curve (AUC), bootstrap resampling, and decision curve analysis. External validation was performed using 171 LGD cases. Statistical techniques such as logistic regression and resampling methods were employed to assess the models' predictive abilities and robustness. Results: In the training dataset, the traditional model achieved an AUC of 0.824 (95% confidence interval [CI]: 0.783-0.865) for predicting pathological upgrading. However, the enhanced model, which incorporated AMACR staining, exhibited a significantly improved performance with an AUC of 0.878 (95% CI: 0.843-0.913). This increase in AUC by 0.054 (95% CI: 0.015-0.093) demonstrates a statistically significant enhancement provided by the inclusion of AMACR staining in the prediction model for pathological upgrading of LGD lesions in gastric mucosa. Conclusion: The findings of this study highlight the practical implications of the enhanced prediction model incorporating AMACR staining for low-grade gastric mucosal dysplasia (LGD). The significantly improved performance of the enhanced model in predicting pathological upgrading emphasizes its potential to revolutionize the management and treatment strategies for patients with LGD. By providing a more accurate prediction of upgrading, the enhanced model enables early intervention and timely decision-making, leading to improved outcomes and prognosis for patients. The incorporation of AMACR staining in the prediction model holds promise for enhancing diagnostic strategies and reducing the incidence of postoperative pathological upgrading. This research underscores the importance of leveraging advanced techniques to improve the early detection rate of gastric cancer and ultimately benefit patient care.
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Background: We aimed to compare the prevention of hypoxemia using High-flow nasal oxygen (HFNO) or regular nasal tubing (CNC) in elderly patients undergoing gastroscopy with sedation. Methods: This study was a prospective, randomized, controlled trial conducted at a single center. We included elective patients aged 65 and above who were undergoing gastroscopy with sedation. In the intervention group (HFNO), we set the oxygen flow rate to 60 liters per minute with an oxygen fraction (FiO2) of 0.6, while in the control group (CNC), it was 6 liters per minute. The primary outcome was the occurrence of hypoxemia (defined as Spo2 < 90%). Results: A total of 125 participants were enrolled (HFNO group: n = 63; CNC group: n = 62). The occurrence of hypoxemia was found to be significantly lower in the HFNO group compared to the CNC group (3.2% vs. 22.6%, p = 0.001). Additionally, a significantly shorter duration of low oxygen levels was observed in the HFNO group [0.0 seconds (0.0-13.0)] compared to the CNC group [0.0 seconds (0.0-124.0), p<0.001]. Moreover, a higher minimum Spo2 value was achieved in the HFNO group [99.0% (98.0-100.0) vs. 96.5% (91.0-99.0), p < 0.001], and a shorter recovery time was recorded [0.5 minutes (0.0-0.5) vs. 0.5 minutes (0.0-1.0), p = 0.016] in comparison to the CNC group. There were no differences in terms of comfort level [0 (0-4) vs. 0 (0-5), p = 0.268] between the two groups. Conclusions: The HFNO system was determined to be a safe and highly effective method for oxygen delivery, leading to a reduction in the occurrence of hypoxemia in elderly patients undergoing gastroscopy with sedation. It is recommended that HFNO be considered as the standard approach for management in this population.
Subject(s)
Gastroscopy , Oxygen , Aged , Humans , Gastroscopy/adverse effects , Cannula , Prospective Studies , Hypoxia/etiology , Hypoxia/prevention & controlABSTRACT
Background/Aims: : Ulcerative colitis (UC) is an incurable, relapsing-remitting inflammatory disease that increases steadily. Mucosal healing has become the primary therapeutic objective for UC. Nevertheless, endoscopic assessments are invasive, expensive, time-consuming, and inconvenient. Therefore, it is crucial to develop a noninvasive predictive model to monitor endoscopic activity in patients with UC. Methods: : Clinical data of 198 adult patients with UC were collected from January 2016 to August 2022 at Huadong Hospital, China. Results: : Patients with UC were randomly divided into the training cohort (70%, n=138) and the validation cohort (30%, n=60). The receiver operating characteristic curve value for the training group was 0.858 (95% confidence interval [CI], 0.781 to 0.936), whereas it was 0.845 (95% CI, 0.731 to 0.960) for the validation group. The calibration curve employed the Hosmer-Lemeshow test (p>0.05) to demonstrate the consistency between the predicted and the actual probabilities in the nomogram of these two groups. The decision curve analysis validated that the nomogram had clinical usefulness. Conclusions: : The nomogram, which incorporated activated partial thromboplastin time, fecal occult blood test, ß2-globulin level, and fibrinogen degradation products, served as a prospective tool for evaluating UC activity in clinical practices.
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Inflammatory signals from immunological cells may cause damage to intestinal epithelial cells (IECs), resulting in intestinal inflammation and tissue impairment. Interferon-γ-inducible protein 16 (IFI16) was reported to be involved in the pathogenesis of Behçet's syndrome (BS). This study aimed to investigate how inflammatory cytokines released by immunological cells and IFI16 participate in the pathogenesis of intestinal BS. RNA sequencing and real-time quantitative PCR (qPCR) showed that the positive regulation of tumor necrosis factor-α (TNF-α) production in peripheral blood mononuclear cells (PBMCs) of intestinal BS patients may be related to the upregulation of polo like kinase 1 (PLK1) in PBMCs (P = 0.012). The plasma TNF-α protein level in intestinal BS was significantly higher than in healthy controls (HCs; P = 0.009). PBMCs of intestinal BS patients and HCs were co-cultured with human normal IECs (NCM460) to explore the interaction between immunological cells and IECs. Using IFI16 knockdown, PBMC-NCM460 co-culture, TNF-α neutralizing monoclonal antibody (mAb), stimulator of interferon genes (STING) agonist 2'3'-cGAMP, and the PLK1 inhibitor SBE 13 HCL, we found that PLK1 promotes the secretion of TNF-α from PBMCs of intestinal BS patients, which causes overexpression of IFI16 and induces apoptosis of IECs via the STING-TBK1 pathway. The expressions of IFI16, TNF-α, cleaved caspase 3, phosphorylated STING (pSTING) and phosphorylated tank binding kinase 1 (pTBK1) in the intestinal ulcer tissue of BS patients were significantly higher than that of HCs (all P < 0.05). PLK1 in PBMCs of intestinal BS patients increased TNF-α secretion, inducing IEC apoptosis via activation of the IFI16-STING-TBK1 pathway. PLK1 and the IFI16-STING-TBK1 pathway may be new therapeutic targets for intestinal BS.
Subject(s)
Apoptosis , Behcet Syndrome , Cell Cycle Proteins , Membrane Proteins , Phosphoproteins , Polo-Like Kinase 1 , Protein Serine-Threonine Kinases , Proto-Oncogene Proteins , Signal Transduction , Humans , Behcet Syndrome/pathology , Behcet Syndrome/metabolism , Behcet Syndrome/genetics , Protein Serine-Threonine Kinases/metabolism , Protein Serine-Threonine Kinases/genetics , Proto-Oncogene Proteins/metabolism , Proto-Oncogene Proteins/genetics , Male , Cell Cycle Proteins/genetics , Cell Cycle Proteins/metabolism , Female , Adult , Membrane Proteins/genetics , Membrane Proteins/metabolism , Phosphoproteins/metabolism , Phosphoproteins/genetics , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/pathology , Epithelial Cells/metabolism , Epithelial Cells/pathology , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factor-alpha/genetics , Intestinal Mucosa/pathology , Intestinal Mucosa/metabolism , Middle AgedABSTRACT
Aim: This study established a high-throughput multiplex genetic detection assay (HMGA) for rapid identification, semi-quantification and virulence analysis of Helicobacter pylori directly from the clinical non-invasive oral samples. Methods: The gastric mucosa and oral samples were collected from 242 patients in Shanghai from 2021 to 2022. All the samples were detected by routine clinical tests for H. pylori and Sanger sequenced for inconsistent results. A new multiplex PCR assay providing results within 4 hours was designed and optimized involving fluorescent dye-labeled specific primers targeted 16S rRNA gene, semi-quantitative gene ureC and 10 virulence genes of H. pylori. Semi-quantification was carried out by simulating the serial 10-fold dilutions of positive oral samples, and the H. pylori loads in different clinical samples were further compared. The mixed plasmids of virulence genes vacA s1, vacA m1 and vacA m2 were used to evaluate the performance on different genotypes. The consistency of 10 virulence genes in gastric mucosa, saliva, mouthwash and dental plaque of H. pylori-positive patients was compared. Results: The non-invasive HMGA was highly specific for detection of all 12 targets of H. pylori and human internal reference gene ß-globin, and the sensitivity to all target genes could reach 10 copies/µL. Compared with routine clinical tests and sequencing, non-invasive HMGA has a high level (>0.98) of sensitivity, specificity, accuracy, PPV, NPV and kappa coefficient for direct detection of H. pylori in oral samples. Moreover, by detecting peak area levels of ureC, it was confirmed that the H. pylori loads in gastric mucosa were significantly higher than those of the three kinds of oral samples (p<0.05). We also found that 45.0% (91/202) of patients had different H. pylori virulence genes in different oral samples. The concordance of positive detection rates of each virulence gene between saliva and gastric mucosa was more than 78% (p<0.05). Conclusion: The non-invasive HMGA proved to be a reliable method for the rapid H. pylori identification, semi-quantification and detection of 10 virulence genes directly in oral samples, providing a new idea for non-invasive detection of H. pylori.
Subject(s)
HMGA Proteins , Helicobacter Infections , Helicobacter pylori , Humans , Bacterial Proteins/genetics , Virulence/genetics , Genotype , RNA, Ribosomal, 16S/genetics , China , HMGA Proteins/genetics , Helicobacter Infections/diagnosis , Antigens, Bacterial/geneticsABSTRACT
BACKGROUND: Shanghai, in east China, has one of the world's highest burdens of Helicobacter pylori infection. While multidrug regimens can effectively eradicate H. pylori, the increasing prevalence of antibiotic resistance (AR) in H. pylori has been recognized by the WHO as 'high priority' for urgent need of new therapies. Moreover, the genetic characteristics of H. pylori AR in Shanghai is under-reported. The purpose of this study was to determine the resistance prevalence, re-substantiate resistance-conferring mutations, and investigate novel genetic elements associated with H. pylori AR. RESULTS: We performed whole genome sequencing and antimicrobial susceptibility testing of 112 H. pylori strains isolated from gastric biopsy specimens from Shanghai patients with different gastric diseases. No strains were resistant to amoxicillin. Levofloxacin, metronidazole and clarithromycin resistance was observed in 39 (34.8%), 73 (65.2%) and 18 (16.1%) strains, respectively. There was no association between gastroscopy diagnosis and resistance phenotypes. We reported the presence or absence of several subsystem protein coding genes including hopE, hofF, spaB, cagY and pflA, and a combination of CRISPRs, which were potentially correlated with resistance phenotypes. The H. pylori strains were also annotated for 80 genome-wide AR genes (ARGs). A genome-wide ARG analysis was performed for the three antibiotics by correlating the phenotypes with the genetic variants, which identified the well-known intrinsic mutations conferring resistance to levofloxacin (N87T/I and/or D91G/Y mutations in gyrA), metronidazole (I38V mutation in fdxB), and clarithromycin (A2143G and/or A2142G mutations in 23S rRNA), and added 174 novel variations, including 23 non-synonymous SNPs and 48 frameshift Indels that were significantly enriched in either the antibiotic-resistant or antibiotic-susceptible bacterial populations. The variant-level linkage disequilibrium analysis highlighted variations in a protease Lon with strong co-occurring correlation with a series of resistance-associated variants. CONCLUSION: Our study revealed multidrug antibiotic resistance in H. pylori strains from Shanghai, which was characterized by high metronidazole and moderate levofloxacin resistance, and identified specific genomic characteristics in relation to H. pylori AR. Continued surveillance of H. pylori AR in Shanghai is warranted in order to establish appropriate eradication treatment regimens for this population.
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Helicobacter pylori is a human pathogen competent for natural transformation. Intrinsic and acquired antibiotic resistance contribute to the survival and multiplication of H. pylori under antibiotics. While drug-resistance dissemination by natural transformation (NT)-mediated horizontal gene transfer remains poorly understood in H. pylori. The purpose of the study was to investigate the role of H. pylori porins (HopA, HopB, HopC, HopD, and HopE) in the intrinsic antibiotic resistance and to preliminarily reveal the potential effect of HopE and HopD porins in streptomycin resistance acquisition after NT in the presence of antibiotics. Using traditional antibiotic susceptibility tests and growth curve analysis, we found the MIC values of metronidazole, clarithromycin, levofloxacin, tetracycline, rifampin, and streptomycin in mutants lacking HopE and/or HopD were significantly elevated compare to those in wild-type strain. The quantitative analysis of the tetramethyl rhodamine isothiocyanate (TRITC)-labeled streptomycin accumulation at the single-cell level showed reduced streptomycin intracellular fluorescence in ΔhopE and ΔhopD mutant cells. Furthermore, in the presence of translation-inhibiting antibiotic streptomycin, the resistance acquisition frequency was decreased in the wild-type strain, which could be reversed by mutants lacking HopE and HopD that restored relatively high resistance acquisition frequencies. By transforming a pUC19-rpsLmut-sfgfp linear plasmid carrying a streptomycin conferring mutation, we observed that the impaired ability of rpsLmut synthesis in the wild-type strain was restored in the ΔhopE and ΔhopD mutant transformants. Our study revealed that in the presence of streptomycin, resistance acquisition at least partially relied on the deletion of the hopE and hopD genes, because their loss reduced streptomycin concentration in the cell and thus restored the expression of the resistance-conferring gene, which was inhibited by streptomycin in wild-type strain. The loss of HopE and HopD influx activity may also preserve resistance acquisition by transformation in the presence of antibiotics with other modes of action. IMPORTANCE Helicobacter pylori is constitutively competent for natural transformation (NT) and possesses an efficient system for homologous recombination, which could be utilized to study the NT-mediated horizontal gene transfer induced antibiotic resistance acquisition. Bacterial porins have drawn renewed attention because of their crucial role in antibiotic susceptibility. From the perspective of porin-mediated influx in H. pylori, our study preliminarily revealed the important role of HopE and HopD porins not only in preserving the intrinsic susceptibility to specific antibiotic but also in evading acquired antibiotic resistance by NT in the presence of translation-inhibiting antimicrobial. Therefore, the loss of HopE or HopD porin in H. pylori genomes, combined with the large number of secreted or cell-free genetic elements carrying mutations conferring antibiotic resistance, may raise the possibility that this mechanism plays a potential role in the propagation of antibiotic resistance within H. pylori communities.
Subject(s)
Anti-Infective Agents , Helicobacter Infections , Helicobacter pylori , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/therapeutic use , Anti-Infective Agents/pharmacology , Drug Resistance, Bacterial/genetics , Helicobacter pylori/genetics , Humans , Microbial Sensitivity Tests , Porins/genetics , Porins/pharmacology , Streptomycin/pharmacologyABSTRACT
BACKGROUND: The current international consensus report indicated that all Helicobacter pylori (H. pylori)-positive patients should be treated. This study aimed to evaluate the long-term effects and benefits of H. pylori eradication on the gastric mucosa in the elderly population. METHODS: We performed a retrospective cohort study with 311 individuals aged ≥60 years, including 83 with persistent H. pylori infection (persistent group), 128 with successful H. pylori eradication (eradicated group), and 100 without H. pylori infection (control group). The results of endoscopy and mucosal histology were investigated at baseline and followed up for 5 and 10 years. RESULTS: In the 5 to 10-year follow-up, there was a significant difference in the atrophy score among the three groups (P < 0.001); however, no significant difference was observed in the intestinal metaplasia (IM) score (P > 0.05). There was no significant difference in the cumulative incidence of gastric neoplastic lesion (GNL) between the eradicated and persistent groups during the 5 to 10-year follow-up period (P > 0.05). The baseline IM score of patients with GNL was significantly higher than that of those without GNL in the eradicated and control groups (P < 0.05). In all patients with GNL, the mean interval time between baseline and diagnosis of GLN was more than 6 years. The severity of baseline mucosal IM (odds ratio: OR 3.092, 95% confidence interval [CI]: 1.690-5.655, P < 0.001) and H. pylori infection (OR: 2.413, 95%CI: 1.019-5.712, P = 0.045) significantly increased the risk for GNL. CONCLUSIONS: Older patients with a life expectancy of less than 5 to 10 years, especially those with moderate to severe gastric mucosal IM, may not benefit from the eradication of H. pylori to prevent gastric cancer.
Subject(s)
Helicobacter Infections , Helicobacter pylori , Precancerous Conditions , Stomach Neoplasms , Aged , Anti-Bacterial Agents/therapeutic use , Follow-Up Studies , Gastric Mucosa/pathology , Helicobacter Infections/drug therapy , Helicobacter Infections/epidemiology , Helicobacter Infections/pathology , Humans , Metaplasia/drug therapy , Metaplasia/pathology , Precancerous Conditions/diagnosis , Precancerous Conditions/drug therapy , Precancerous Conditions/pathology , Retrospective Studies , Stomach Neoplasms/epidemiology , Stomach Neoplasms/pathology , Stomach Neoplasms/prevention & controlABSTRACT
BACKGROUND: With the development of new technologies such as magnifying endoscopy with narrow band imaging, endoscopists achieved better accuracy for diagnosis of gastric cancer (GC) in various aspects. However, to master such skill takes substantial effort and could be difficult for inexperienced doctors. Therefore, a novel diagnostic method based on artificial intelligence (AI) was developed and its effectiveness was confirmed in many studies. AI system using convolutional neural network has showed marvelous results in the ongoing trials of computer-aided detection of colorectal polyps. SUMMARY: With AI's efficient computational power and learning capacities, endoscopists could improve their diagnostic accuracy and avoid the overlooking or over-diagnosis of gastric neoplasm. Several systems have been reported to achieved decent accuracy. Thus, AI-assisted endoscopy showed great potential on more accurate and sensitive ways for early detection, differentiation, and invasion depth prediction of gastric lesions. However, the feasibility, effectiveness, and safety in daily practice remain to be tested. Key messages: This review summarizes the current status of different AI applications in early GC diagnosis. More randomized controlled trails will be needed before AI could be widely put into clinical practice.
Subject(s)
Stomach Neoplasms , Artificial Intelligence , Endoscopy, Gastrointestinal , Humans , Narrow Band Imaging , Overdiagnosis , Stomach Neoplasms/diagnostic imagingABSTRACT
Syphilis is a chronic systemic infectious disease. Gastric involvement is uncommon. We report of a 55-year-old female patient with a history of epigastric pain and a diagnosis of Syphilis confirmed by main clinical symptoms, tolulized red unheated serum test(TRUST) and treponema pallidum hemagglutination assay(TPHA). At the same time, we still reported gastric involvement about syphilis in this patient with gastroscopy and pathology.
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AIMS: To compare the efficacy and the safety of low-dose (3.5 mg/kg) or standard-dose (5 mg/kg) of infliximab (IFX) for induction and maintenance treatment in Chinese patients with intestinal Behcet's disease (BD), and to identify potential predictors to corticosteroid-free clinical remission at week 30. METHODS: A prospective trial was conducted in 20 patients with moderate-to-severe active intestinal BD, followed up for 30 weeks. Ileocolonoscopic examinations were done at baseline and week 14. A logistic regression model was used to assess the predictors of corticosteroid-free remission at week 30. RESULTS: As the primary end point, steroid-free remission at week 30 was observed in 40% and 60% of patients who received low and standard doses of IFX, respectively (P>0.371). As the secondary end point, the mucosal healing (MH) rates at week 14 were found to be 60% for both low and standard doses of IFX. The clinical response rates at week 14 and 30 were also similar in both groups. Only MH at week 14 predicted steroid-free clinical remission at week 30. CONCLUSIONS: Low-dose of IFX is effective and safe for induction and maintenance therapy in patients with active intestinal BD. MH at week 14 predicted clinical remission at week 30.
Subject(s)
Behcet Syndrome/drug therapy , Behcet Syndrome/pathology , Infliximab/therapeutic use , Intestinal Mucosa/pathology , Wound Healing , Adolescent , Adult , Aged , Asian People , Demography , Dose-Response Relationship, Drug , Endpoint Determination , Female , Humans , Infliximab/administration & dosage , Infliximab/adverse effects , Male , Middle Aged , Remission Induction , Treatment Outcome , Young AdultABSTRACT
BACKGROUND: Intestinal Behcet's disease (BD) is a specific subtype of BD. Effective drug therapy for intestinal BD remains elusive. AIMS: To investigate long-term outcomes and identify predictors of sustained response in intestinal BD patients receiving infliximab (IFX) treatment. METHODS: The medical records were reviewed of patients received IFX from September 2012 to March 2016. The cumulative probabilities of sustained response were calculated using the Kaplan-Meier. Predictor factors for sustained response were accessed by receiver operating characteristic curve. RESULTS: Totally, 27 active intestinal BD patients were enrolled. Sustained responses were observed in 17 patients, after a median follow-up duration 24 months (interquartile range 9-37). The proportion of clinical remission at week 14, 30, and 52 had occurred in 84.6, 70, and 70%, respectively, with the proportion of clinical remission of 69.2, 40, and 55%. The mucosal healing (MH) rate at week 14 was 72%. Kaplan-Meier estimated patients with achievement of clinical and biological responses at week 14 or MH was likely to remain sustained clinical response. ROC curve analysis revealed CRP level (of 6.85 mg/L) at week 14 is a potential predictor for discriminating patients with sustained response from relapse, with an area under the curve values of 0.837. CONCLUSIONS: IFX is effective and safe for induction and maintenance therapy in Chinese patients with moderate-to-severe active intestinal BD. Early achievement of clinical response and mucosal healing might associate long-term response. A lower CRP level seems to be associated with a more benign clinical course.
Subject(s)
Behcet Syndrome/drug therapy , Gastrointestinal Agents/therapeutic use , Infliximab/therapeutic use , Intestinal Diseases/drug therapy , Adrenal Cortex Hormones/therapeutic use , Adult , Behcet Syndrome/complications , Behcet Syndrome/metabolism , C-Reactive Protein/metabolism , China , Drug Therapy, Combination , Female , Humans , Immunosuppressive Agents/therapeutic use , Intestinal Diseases/etiology , Intestinal Diseases/metabolism , Kaplan-Meier Estimate , Maintenance Chemotherapy , Male , Prognosis , ROC Curve , Remission Induction , Retrospective Studies , Treatment Outcome , Young AdultABSTRACT
AIMS: To examine the expression of miRNAs and mRNAs of heme oxygenase 1 (HO-1) in patients with active intestinal Behcet's disease (BD). METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from BD patients with active ileocecal ulcers or from healthy donors. Expression levels of four miRNAs were studied using real-time PCR. The levels of mRNAs of ho-1, bach1, and CD14 were measured by qRT-PCR. Serum levels of cytokines were analyzed by ELISA. RESULTS: Among four miRNAs, only levels of miR-196a2 were significantly decreased from BD patients with active ileocecal ulcers as compared with healthy controls. Moreover, level of mRNA ho-1 expression in PBMCs from patients with BD was reduced. No significant difference on bach1 and CD14 mRNA levels was observed. Levels of IFN-γ, IL-17, IL-10, IL-1ß, and TNF-α were higher in patients with active intestinal BD than those in healthy controls. CONCLUSION: The present results suggest that miR-196a2 expression is decreased in active intestinal BD patients. Down regulated miR-196a2 may be involved in intestinal BD pathogenesis by targeting Bach1/ho-1. Consequently, pro-inflammatory cytokines are closely implicated in the evolution of intestinal BD.
Subject(s)
Behcet Syndrome/enzymology , Behcet Syndrome/genetics , Gene Expression Regulation , Heme Oxygenase-1/metabolism , Leukocytes, Mononuclear/metabolism , MicroRNAs/genetics , Adult , Basic-Leucine Zipper Transcription Factors/genetics , Basic-Leucine Zipper Transcription Factors/metabolism , Behcet Syndrome/blood , Cytokines/blood , Fanconi Anemia Complementation Group Proteins/genetics , Fanconi Anemia Complementation Group Proteins/metabolism , Female , Heme Oxygenase-1/genetics , Humans , Male , MicroRNAs/metabolism , RNA, Messenger/genetics , RNA, Messenger/metabolismABSTRACT
Helicobacter pylori (H. pylori) infection is closely related to various gastroduodenal diseases. Virulence factors and bacterial load of H. pylori are associated with clinical outcomes, and drug-resistance severely impacts the clinical efficacy of eradication treatment. Existing detection methods are low-throughput, time-consuming and labor intensive. Therefore, a rapid and high-throughput method is needed for clinical diagnosis, treatment, and monitoring for H. pylori. High-throughput Multiplex Genetic Detection System (HMGS) assay was established to simultaneously detect and analyze a set of genes for H. pylori identification, quantification, virulence, and drug resistance by optimizing the singlet-PCR and multiple primers assay. Twenty-one pairs of chimeric primers consisted of conserved and specific gene sequences of H. pylori tagged with universal sequence at the 5' end were designed. Singlet-PCR assay and multiple primers assay were developed to optimize the HMGS. The specificity of HMGS assay was evaluated using standard H. pylori strains and bacterial controls. Six clinical isolates with known genetic background of target genes were detected to assess the accuracy of HMGS assay. Artificial mixed pathogen DNA templates were used to evaluate the ability to distinguish mixed infections using HMGS assay. Furthermore, gastric biopsy specimens with corresponding isolated strains were used to assess the capability of HMGS assay in detecting biopsy specimens directly. HMGS assay was specific for H. pylori identification. HMGS assay for H. pylori target genes detection were completely consistent with the corresponding genetic background. Mixed infection with different drug-resistant isolates of H. pylori could be distinguished by HMGS assay. HMGS assay could efficiently diagnose H. pylori infection in gastric biopsy specimens directly. HMGS assay is a rapid and high throughput method for the simultaneous identification and quantification of H. pylori, analysis of virulence and drug resistance in both isolated strains and biopsy specimens. It could also be used to distinguish the mixed infection with different resistant genotype strains. Furthermore, HMGS could detect H. pylori infection in gastric biopsy specimens directly.
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AIM: We evaluated the direct high-throughput multiple genetic detection system (dHMGS) for Helicobacter pylori in gastric biopsies. MATERIALS & METHODS: One hundred and thirty-three specimens were concurrently analyzed by dHMGS, rapid urease test, culture and sequencing. RESULTS: dHMGS was highly sensitive and specific for H. pylori identification compared with culture and rapid urease test. The correlation coefficient of the quantitative standard curve was R2 = 0.983. A significant difference in the relative H. pylori DNA abundance was found in different gastroduodenal diseases. Concordance rates between dHMGS and sequencing for resistance mutations were 97.1, 100.0, 85.3 and 97.1%, respectively. Finally, dHMGS could efficiently distinguish mixed infection in biopsy specimens. CONCLUSION: The dHMGS could efficiently diagnose and quantify H. pylori burden in biopsies, simultaneously screening for virulence, antibiotic resistance and presence of the multistrain infections.
Subject(s)
Helicobacter Infections/microbiology , Helicobacter pylori/isolation & purification , High-Throughput Nucleotide Sequencing/methods , Adult , Aged , Aged, 80 and over , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Biopsy , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , Female , Helicobacter Infections/pathology , Helicobacter pylori/classification , Helicobacter pylori/genetics , Humans , Male , Middle Aged , Polymerase Chain Reaction , Sensitivity and Specificity , Urease/genetics , Urease/metabolism , Young AdultABSTRACT
Antibiotics resistance in Helicobacter pylori (H. pylori) is the major factor for eradication failure. Molecular tests including fluorescence in situ hybridization, PCR-restriction fragment length polymorphism, and dual priming oligonucleotide-PCR (DPO-PCR) play critical roles in the detection of antibiotic susceptibility; however, limited knowledge is known about application of multiple genetic analysis system (MGAS) in the area of H. pylori identification and antibiotics resistance detection.The aim of this study is to determine the antibiotics resistance using different molecular tests and evaluate the treatment outcomes of E-test-based genotypic resistance.A total of 297 patients with dyspepsia complaint were recruited for gastroscopies. Ninety patients with H. pylori culture positive were randomly divided into 2 groups (test group and control group). E-test, general PCR, and MGAS assay were performed in test group. Patients in control group were treated with empirical therapy (rabeprazole + bismuth potassium citrate + amoxicillin [AMX] + clarithromycin [CLR]), whereas patients in test group received quadruple therapy based on E-test results twice daily for 14 consecutive days. The eradication effect of H. pylori was confirmed by C-urea breath test after at least 4 weeks when treatment was finished.Rapid urease test showed 46.5% (128/297) patients with H. pylori infection, whereas 30.3% (90/297) patients were H. pylori culture positive. E-test showed that H. pylori primary resistance rate to CLR, AMX, metronidazole, tetracycline, and levofloxacin (LVX) was 40.0% (18/45), 4.4% (2/45), 53.3% (24/45), 0% (0/45), and 55.6% (25/45), respectively. In addition, there are many multidrug resistant (MDR) phenotypes, and the MDR strains have higher minimum inhibitory concentration than their single-drug resistant counterparts. Considering E-test as the reference test, the sensitivities of general PCR and MGAS in detecting CLR resistance were 83.3% (15/18) and 94.4% (17/18), whereas in detecting LVX resistance were 100% (25/25) and 83.3% (15/18), respectively. Finally, the eradication rate in test group was significantly higher than that in control group as demonstrated by intention-to-treat analysis and per-protocol analysis.MGAS is a promising assay for H. pylori identification and antibiotic susceptibility testing. Phenotypic resistance-guided quadruple therapy showed a high efficacy in treating patients with H. pylori infection.
Subject(s)
Anti-Bacterial Agents , Drug Resistance, Microbial/genetics , Dyspepsia , Genome, Bacterial , Helicobacter Infections , Helicobacter pylori , Adult , Anti-Bacterial Agents/classification , Anti-Bacterial Agents/pharmacology , Breath Tests/methods , Drug Monitoring , Drug Therapy, Combination/methods , Dyspepsia/diagnosis , Dyspepsia/drug therapy , Dyspepsia/microbiology , Female , Helicobacter Infections/diagnosis , Helicobacter Infections/drug therapy , Helicobacter Infections/microbiology , Helicobacter pylori/drug effects , Helicobacter pylori/genetics , Helicobacter pylori/isolation & purification , Humans , Male , Microbial Sensitivity Tests/methods , Middle Aged , Precision Medicine/methods , Sensitivity and Specificity , Treatment OutcomeABSTRACT
BACKGROUND: Currently, several diagnostic assays for Helicobacter pylori (H. pylori) are available, but each has some limitations. Further, a high-flux quantitative assay is required to assist clinical diagnosis and monitor the effectiveness of therapy and novel vaccine candidates. METHODS: Three hundred and eighty-seven adult patients [nonulcer dyspepsia (NUD) 295, peptic ulcer disease (PUD) 77, gastric cancer (GC) 15] were enrolled for gastrointestinal endoscopies. Three biopsy samples from gastric antrum were collected for the following tests: culture, rapid urease test (RUT), histopathology, conventional polymerase chain reaction (PCR), and Multiple Genetic Analysis System (MGAS). The diagnostic capability of H. pylori for all methods was evaluated through the receiver operating characteristic (ROC) curves. RESULTS: Based on the gold standard, the sensitivity and specificity of MGAS were 92.9 and 92.4%, and positive predict value (PPV) and negative predict value (NPV) were 96.0 and 87.1%, respectively. All the above parameters of MGAS were higher than that of culture (except its specificity), RUT and histopathology, and nearly closed to that of conventional PCR. The area under curve (AUC) was 0.7575 (Culture), 0.8870 (RUT), 0.9000 (Histopathology), 0.9496 (Conventional PCR), and 0.9277 (MGAS). No significant statistical difference was observed for the H. pylori DNA load in different disease groups (p = .067). In contrast, a statistically significant difference in the H. pylori DNA copy number was observed based on age (p = .043) and gender (p = .021). CONCLUSIONS: The data showed that MGAS performed well in detecting H. pylori infection. Furthermore, the quantitative analysis showed that the load of H. pylori was significantly different within both age and gender groups. These results suggested that MGAS could be a potential alternative method for clinical detection and monitoring of the effectiveness of H. pylori therapy.
Subject(s)
Bacteriological Techniques/methods , Diagnostic Tests, Routine/methods , Helicobacter Infections/diagnosis , Helicobacter pylori/isolation & purification , Histocytochemistry/methods , Molecular Diagnostic Techniques/methods , Adult , Aged , Aged, 80 and over , Biopsy , Endoscopy, Gastrointestinal , Female , Gastric Mucosa/microbiology , Gastric Mucosa/pathology , Humans , Male , Middle Aged , Predictive Value of Tests , Sensitivity and Specificity , Young AdultABSTRACT
AIM: To report the incidence, clinical features and outcomes of gastrointestinal (GI) involvement in Behcet's disease (BD). METHODS: A total of 168 consecutive patients with BD were screened and upper and lower GI endoscopies were performed in 148 patients. Four hundred age- and sex-matched controls were enrolled for comparison. RESULTS: Fifty-two (35.1%) patients had GI lesions. After a mean follow-up of 10 mo, ileocecal ulcers had been confirmed in 20 patients, including active ulcer(s) in 18 patients, but no ileocecal ulceration was found in controls. GI symptoms were present in 14 patients with active ulcer(s), while 4 patients with smaller ulcer were asymptomatic. Endoscopic features of ileocecal ulcer were: a single ulcer (50%), larger than 1 cm in diameter (72.2%), and round/oval or volcano-type in shape (83.3%). Compared with patients without GI involvement, less ocular lesions, lower levels of albumin, erythrocyte count and hemoglobin, and higher levels of C-reactive protein and erythrocyte sedimentation rate were confirmed in the intestinal BD group. Four patients had esophageal ulcers in the BD group but no case in controls. The other endoscopic findings were similar between the two groups. The prevalence of Helicobacter pylori infection was similar in both groups. Most patients received an immunomodulator and responded well. CONCLUSION: GI lesions commonly occur in Chinese BD patients. The most frequently involved area is the ileocecal region. Esophageal ulcer might be a rare but unique lesion.