Comprehensive analysis of transcriptome-wide N6-methyladenosine methylomes in the Barrett's esophagus in rats.

PURPOSE: As the most abundant RNA modification, N6-methyladenosine (m6A) methylation plays crucial roles in various diseases. The aim of this study is to comprehensively map the landscape of the mRNA m6A modification pattern in Barrett's esophagus (BE) in order to find key genes and potential therapy for BE and even esophageal adenocarcinoma (EAC). METHODS: Methylated RNA immunoprecipitation sequencing (MeRIP-seq) and RNA-sequencing (RNA-seq) were performed to compare the difference in mRNA m6A methylation and differentially expressed mRNAs between BE and normal control (NC) tissues. Bioinformatics analysis was used to describe the m6A modification pattern and specific genes in BE and NC tissues. RESULTS: Through MeRIP-seq, we obtained m6A methylation profiling in BE and NC tissues. In total, 11,026 unique peaks were detected in the BE groups, whereas 8564 unique peaks were detected in the NC groups. Peaks were primarily enriched within CDS with GGACU motifs and most of the peaks were within 1000 bp in width. Moreover, functional enrichment analysis demonstrated that hypermethylated and hypomethylated genes were significantly enriched in coronavirus disease pathway, calcium signaling pathway and MAPK signaling pathways. Furthermore, PPI network was conducted and 18 hub genes were identified via STRING database and Cystoscope. Among them, ACTA1, CDC20, CKM, KIF20a, MYH11, TPM2, MYL9, DES, TNNT3 were overexpressed in EAC in the GEPIA gene bank and TPM1, KIF20a impaired patients' survival in the Kaplan-Meier plotter database. Finally, functional enrichment analysis demonstrated that co-expressed genes of TPM1 were significantly enriched in calcium signaling pathway, cGMP-PKG signaling pathway and PI3K-Akt signaling pathway. CONCLUSION: Our study is the first to perform comprehensive and transcriptome-wide maps to identify the potential roles played by m6A methylation in BE, which widely involved in oxidative stress. This foresees a guiding role in revealing the molecular mechanism of m6A-mediated genes that govern the pathogenesis and progression of BE and EAC.

[Research progress of mesenchymal stem cell-derived exosomes in the treatment of inflammatory bowel disease].

Inflammatory bowel disease (IBD) is a chronic nonspecific inflammatory disease of the intestine, with unknown etiology and the incidence is increasing year by year. Traditional treatment has limited effect. Mesenchymal stem cell-derived exosomes (MSC-Exos) are a group of nano-sized extracellular vesicles. Their function is equivalent to that of mesenchymal stem cells (MSCs), with no tumorigenicity and high safety. They represent a novel cell-free therapy. It has been shown that MSC-Exos can improve IBD by effects including anti-inflammation, antioxidant stress, repairing intestinal mucosal barrier and immune regulation. However, their clinical application still faces some problems, such as the lack of standardized production technology, lack of specific IBD diagnostic molecules and anti-intestinal fibrosis.

Concurrent alcoholic cirrhosis and malignant peritoneal mesothelioma in a patient: A case report.

BACKGROUND: Malignant peritoneal mesothelioma (MPM) originates from the mesothelial and subcutaneous cells of the abdominal cavity. Its diagnose is difficult due to its nonspecific and vague symptoms, and it should be differentiated from alcoholic cirrhosis and liver and pancreatic cancers. Misdiagnosis and missed diagnosis can easily occur when MPM presents with other diseases. To the best of our knowledge, no case of MPM concurrent with alcoholic cirrhosis has been reported. CASE SUMMARY: A 63-year-old man presented to our hospital with abdominal distension for 20days. He had a history of alcohol consumption for nearly 30 years and no history of special drug use or toxic exposure. After treatment for alcoholic cirrhosis in a community hospital, his symptoms did not improve significantly. The patient underwent exploratory laparotomy and surgical resection. Pathologic examination showed an epithelioid MPM. He was treated with chemotherapy and intraperitoneal hyperthermic perfusion after surgery. Currently, he is in a stable condition and tumor recurrence has not occurred. CONCLUSION: Misdiagnosis and missed diagnosis of MPM can easily occur because of its insidious onset. Therefore, there is a need to understand. MPM in clinical practice, make the correct diagnosis, and provide timely and effective treatment.

Ambulatory 24-hour multichannel intraluminal impedance-pH monitoring and high resolution endoscopy distinguish patients with non-erosive reflux disease from those with functional heartburn.

AIMS: To assess the contribution of 24-h esophageal multichannel intraluminal impedance and pH (MII-pH) monitoring and high resolution endoscopy (HRE) with i-scan imaging in differentiating non erosive reflux disease (NERD) from functional heartburn (FH). METHODS: This is a retrospective cohort study of patients with heartburn from the Endoscopy Unit. NERD patients and FH patients were defined by 24-h MII-pH monitoring and white light endoscopy. Minimal mucosal changes were assessed by HRE with i-scan imaging. RESULTS: Total of 156 consecutive patients with heartburn but without esophageal mucosal erosions were studied. Forty-eight of these subjects had NERD, with increased acid exposure time (AET) and positive SAP and/or SI. Eighteen had FH with normal AET and negative SAP and SI. When compared to FH patients and healthy controls, NERD patients had significantly increased number of total acid reflux episodes, as well as increased number of weakly acidic reflux episodes (p<0.01). The rate of proximal reflux episodes in NERD patients was higher than that of FH patients and healthy controls (p<0.01). Irregular or blurring of the Z-line (58.3%) and white mucosal turbidity (47.9%) were the most common endoscopic findings of minimal mucosal changes observed in this study. NERD patients had more prevalent minimal changes than FH patients and the controls (87.5%vs. 66.6%vs. 61.9%; p = 0.004) with sensitivity of 87.5%. Histopathological evaluation showed that NERD patients had significantly higher average scores of intercellular spaces dilation (2.82±0.9 vs. 1.2±0.6, p = 0.005) and papillae elongation (2.65±1.0 vs. 1.5±0.8, p = 0.014), but not for basal cell proliferation (1.6±1.3 vs. 1.0±0.9, p = 0.070). The histological scores of the NERD patients were 7.1±1.2, which were higher than those of FH patients (3.4±1.0, p = 0.004). CONCLUSIONS: Minimal mucosal changes could be useful markers to support clinical diagnosis of NERD. Combination of 24-h MII-pH monitoring and i-scan high resolution endoscopy can distinguish patients with NERD from those with FH.

Imbalance of tumor necrosis factor-α, interleukin-8 and interleukin-10 production evokes barrier dysfunction, severe abdominal symptoms and psychological disorders in patients with irritable bowel syndrome-associated diarrhea.

The present study aimed to explore the correlation between cytokine expression of tumor necrosis factor α (TNF­α), interleukin (IL)­8 and IL­10 with occludin production, abdominal symptoms and psychological factors in patients with irritable bowel syndrome­associated diarrhea (IBS­D). A total of 42 IBS­D patients and 20 healthy controls were included, which were recruited from QiLu Hospital in China. ELISA and immunohistochemical analysis were performed for evaluating the cytokines (TNF­α, IL­8 and IL­10) and occludin protein levels in the peripheral blood mononuclear cells (PBMCs) of all subjects. In addition, the abdominal symptoms and psychological status were assessed in IBS­D patients. Levels of TNF­α and IL­8 in the PBMCs of patients with IBS­D were significantly higher than those in the controls (P<0.001 and P=0.007, respectively), while IL­10 levels were significantly reduced in patients with IBS­D (P=0.047). Occludin production was significantly reduced in patients with IBS­D as compared with that in the controls (P<0.001). In patients with IBS­D, levels of TNF­α and IL­8 were negatively correlated with occludin levels (r=­0.34, P=0.028; r=­0.52, P<0.001, respectively). IL­10 showed a negative correlation with occludin production (r=0.05, P=0.748). Furthermore, TNF­α, IL­8 and IL­10 levels were significantly correlated with symptoms scores (r=0.74, P<0.001; r=0.55, P<0.001; r=­0.80, P<0.001, respectively) in patients with IBS­D. Within the IBS­D group, TNF­α expression was significantly increased in patients with a self­rating depression scale (SDS) score ≥50 (P=0.004) as compared with that in patients with an SDS score <50. Furthermore, IL­8 was significantly increased in IBS­D patients with a self­rating anxiety scale (SAS) or SDS score ≥50 (P=0.016, P=0.008, respectively) as compared with that in patients scoring <50. In conclusion, the results of the present study suggested that in IBS­D, an imbalance of cytokine production evoked colonic epithelial barrier dysfunction, abdominal symptoms and psychological disorders.

Microalterations of esophagus in patients with non-erosive reflux disease: in-vivo diagnosis by confocal laser endomicroscopy and its relationship with gastroesophageal reflux.

OBJECTIVES: Objectively diagnosing non-erosive reflux disease (NERD) is still a challenge. We aimed to evaluate the use of in-vivo confocal laser endomicroscopy (CLE) to examine the microalterations of the esophagus in patients with NERD and its relationship with reflux episodes monitored by multiple intraluminal impedance-pH (MII-pH). METHODS: Patients with gastroesophageal reflux symptoms completed reflux disease questionnaires. NERD was determined by negative gastroscopy. Patients without reflux symptoms were recruited as controls. Pilot clinical study was followed by prospective controlled blinded study. All subjects were examined by white-light mode of the endoscopy followed by the standard CLE mode and then MII-pH monitoring. The microalterations seen on CLE images and the correlation between CLE features and reflux episodes were evaluated, the correlation between CLE and transmission electron microscope (TEM) data was also analyzed. RESULTS: On CLE images, NERD patients had more intrapapillary capillary loops (IPCLs) per image than did controls (8.29 ± 3.52 vs. 5.69 ± 2.31, P=0.010), as well as the diameter of IPCLs (19.48 ± 3.13 vs. 15.87 ± 2.21 µm, P=0.041) and intercellular spaces of squamous cells (3.40 ± 0.82 vs. 1.90 ± 0.53 µm, P=0.042). The receiver operating characteristic analysis indicated that IPCLs number (optimal cutoff >6 per image, area under the curve (AUC) 0.722, 95% confidence interval (CI) 0.592-0.853, sensitivity 67.7%, specificity 71.6%), IPCLs diameter (optimal cutoff >17.2 µm, AUC 0.847, 95% CI 0.747-0.947, sensitivity 81%, specificity 76%), and the intercellular spaces of squamous cells (optimal cutoff >2.40 µm, AUC 0.935, 95% CI 0.875-0.995, sensitivity 85.7%, specificity 90.5%) diagnosed NERD with reasonable accuracy. Combined features of dilatation of intercellular space plus increased IPCLs provided 100% specificity in the diagnosis of NERD patients. The intercellular spaces of squamous cells observed on CLE were highly related to that on TEM findings (r=0.75, P<0.001). Multivariate progressive regression analysis showed that acidic reflux, especially in the supine position, was related to the increased number and dilation of IPCLs in the squamous epithelium (ß=0.063, t=2.895, P=0.038 and ß=0.156, t=1.023, P=0.04). CONCLUSIONS: CLE represents a useful and potentially significant improvement over standard endoscopy to examine the microalterations of the esophagus in vivo. Acidic reflux is responsible for the microalterations in the esophagus of patients with NERD.

Effects on confocal laser endomicroscopy image quality by different acriflavine concentrations.

BACKGROUND: Acriflavine is one of the commonly used staining agents in confocal laser endomicroscopy (CLE), a newly developed technique allows for real time histological observation of gastrointestinal mucosa, but the concentration is not unified. This study aimed to evaluate the effects of acriflavine with different concentrations on the CLE image quality and to find a sound concentration in clinical practice. METHODS: Twenty four consecutive patients who underwent upper gastrointestinal CLE were enrolled into this study. The patients randomly accepted acriflavine in four different concentrations which were the conventional 0.05% and 3 lower ones respectively: 0.02%, 0.01% and 0.005% spraying onto the same focal antrum mucosa during CLE procedures. Differences of Image quality were demonstrated by an objective score system. RESULTS: THERE WAS NO SIGNIFICANT DIFFERENCE ABOUT IMAGE QUALITY AMONG ACRIFLAVINE CONCENTRATIONS: 0.05%, 0.02% and 0.01%, but 0.005% decreased image quality significantly (P=0.012). And 0.005% was also the only one which decreased general assessment significantly (P=0.01). For the 3 diagnostic value assessment indices, there was no significant difference about nonspecific and even staining, while 0.02% showed significant better polar staining (P=0.03). CONCLUSIONS: Acriflavine concentration 0.02% is the best one applied in CLE with the best nuclei staining ability and preserved image quality.

Diagnostic value of confocal laser endomicroscopy for gastric superficial cancerous lesions.

BACKGROUND: The identification of gastric superficial cancerous lesions based on conventional white-light endoscopy (WLE) is challenging, and histological analysis remains the 'gold standard' for the final diagnosis. Confocal laser endomicroscopy (CLE) can provide in vivo histological observation without the need for biopsy. OBJECTIVE: To develop and evaluate CLE imaging criteria for gastric superficial cancerous lesions and to compare the diagnostic value of real-time integrated CLE (iCLE) and WLE alone in distinguishing gastric superficial cancerous lesions. DESIGN: Prospective study. SETTING: Qilu Hospital, Shandong University, Jinan, China. PATIENTS: A total of 182 patients were enrolled into phase I and 1786 patients were enrolled into phase II. INTERVENTIONS: CLE images were blindly evaluated after endoscopy in phase I, and real-time iCLE diagnosis during endoscopy was compared with WLE diagnosis by using histopathology as a gold standard in phase II. MAIN OUTCOME MEASUREMENTS: The validity and reliability of the CLE diagnosis for identifying gastric superficial cancerous lesions. RESULTS: Off-line CLE diagnosis for early gastric cancers had a high sensitivity (88.1%) and specificity (98.6%). When the two-tiered CLE classification of non-cancerous lesions and cancer/high-grade intraepithelial neoplasia (HGIN) lesions was introduced, CLE diagnosis led to a higher sensitivity (90.2%) and specificity (98.5%) (phase I). Real-time iCLE diagnosis had a higher sensitivity (88.9%), specificity (99.3%) and accuracy (98.8%) for gastric superficial cancer/HGIN lesions than WLE diagnosis (sensitivity, 72.2%; specificity, 95.1%; and accuracy, 94.1%) (p < 0.05) (phase II). Limitations This was a single-centre study. CONCLUSIONS: CLE can be used to identify gastric superficial cancer/HGIN lesions with high validity and reliability.