Comparative transcriptomic analysis reveals the molecular changes of acute pancreatitis in experimental models

BACKGROUND Acute pancreatitis(AP)encompasses a spectrum of pancreatic inflammatory conditions,ranging from mild inflammation to severe pancreatic necrosis and multisystem organ failure.Given the challenges associated with obtaining human pancreatic samples,research on AP predominantly relies on animal models.In this study,we aimed to elucidate the fundamental molecular mechanisms underlying AP using various AP models.AIM To investigate the shared molecular changes underlying the development of AP across varying severity levels.METHODS AP was induced in animal models through treatment with caerulein alone or in combination with lipopolysaccharide(LPS).Additionally,using Ptf1αto drive the specific expression of the hM3 promoter in pancreatic acinar cells transgenic C57BL/6J-hM3/Ptf1α(cre)mice were administered Clozapine N-oxide to induce AP.Subsequently,we conducted RNA sequencing of pancreatic tissues and validated the expression of significantly different genes using the Gene Expression Omnibus(GEO)database.RESULTS Caerulein-induced AP showed severe inflammation and edema,which were exacerbated when combined with LPS and accompanied by partial pancreatic tissue necrosis.Compared with the control group,RNA sequencing analysis revealed 880 significantly differentially expressed genes in the caerulein model and 885 in the caerulein combined with the LPS model.Kyoto Encyclopedia of Genes and Genomes enrichment analysis and Gene Set Enrichment Analysis indicated substantial enrichment of the TLR and NOD-like receptor signaling pathway,TLR signaling pathway,and NF-κB signaling pathway,alongside elevated levels of apoptosis-related pathways,such as apoptosis,P53 pathway,and phagosome pathway.The significantly elevated genes in the TLR and NOD-like receptor signaling pathways,as well as in the apoptosis pathway,were validated through quantitative real-time PCR experiments in animal models.Validation from the GEO database revealed that only MYD88 concurred in both mouse pancreatic tissue and human AP peripheral blood,while TLR1,TLR7,RIPK3,and OAS2 genes exhibited marked elevation in human AP.The genes TUBA1A and GADD45A played significant roles in apoptosis within human AP.The transgenic mouse model hM3/Ptf1α(cre)successfully validated significant differential genes in the TLR and NOD-like receptor signaling pathways as well as the apoptosis pathway,indicating that these pathways represent shared pathological processes in AP across different models.CONCLUSION The TLR and NOD receptor signaling pathways play crucial roles in the inflammatory progression of AP,notably the MYD88 gene.Apoptosis holds a central position in the necrotic processes of AP,with TUBA1A and GADD45A genes exhibiting prominence in human AP.

Potential effect of chronic Helicobacter pylori infection on glucose metabolism of Mongolian gerbils

AIM: To assess the effect of Helicobacter pylori(H. pylori) infection on metabolic parameters in Mongolian gerbils.METHODS: A total of 40 male, 5- to 8-wk-old, specific-pathogen-free Mongolian gerbils(30-50 g) were randomly allocated into two groups: a control group(n = 20) and an H. pylori group(n = 20). After a two-week acclimation period, the control group was administered Brucella broth and the H. pylori group was challenged intra-gastrically five times every other day with approximately 109/CFU H. pylori ATCC43504(Cag A+, Vac A+). Each group was then divided into two subgroups, which were sacrificed at either 6 or 12 mo. The control and H. pylori subgroups each contained 10 Mongolian gerbils. Body weight, abdominal circumference, and body length were measured, and body mass index(BMI) and Lee's index were calculated. Biochemical assays were used to detect serum indexes, including glucose, glycated hemoglobin(GHb), glycated hemoglobin A1c(Hb A1c), triacylglycerol, and total cholesterol, using an automatic biochemistry analyzer. Inflammatory cytokines, including interleukin(IL)-1β, IL-2, IL-4,IL-10, IL-12, tumor necrosis factor-α(TNF-α) and interferon(IFN)-g, were assayed using ELISA. The expression of insulin and insulin-like growth factor 1(IGF-1) was detected by immunohistochemistry, and islet apoptosis was measured using the terminal deoxynucleotidyl transferase-mediated d UTP nick end labeling(TUNEL) assay.RESULTS: At each time point, body weight, abdominal circumference, BMI, and Lee's index were increased after H. pylori infection. However, these differences were not significant. H. pylori infection significantly increased the GHb(5.45 ± 0.53 vs 4.98 ± 0.22, P < 0.05) and Hb A1c(4.91 ± 0.61 vs 4.61 ± 0.15, P < 0.05) levels at 12 mo. We observed no significant differences in serum biochemical indexes, including fasting blood glucose, triacylglycerol and total cholesterol, at 6 or 12 mo after infection. H. pylori infection significantly increased the expression of IGF-1(P < 0.05). Insulin levels from the pancreas and the apoptotic rate of islet β-cells remained unchanged. Also, we observed no significant differences among cytokines levels, including IL-1β, IL-2, IL-4, IL-10, IL-12, TNF-α and IFN-g. IL-4 was the only exception, which increased at 6(44.36 ± 25.17 vs 17.38 ± 3.47, P < 0.05) and 12 mo(33.41 ± 10.00 vs 18.91 ± 5.31, P < 0.05) after H. pylori infection.CONCLUSION: Long-term H. pylori infection is significantly associated with high levels of Hb A1 c in Mongolian gerbils, indicating a potential role of H. pylori infection in glucose dysregulation.

内镜下治疗大肠息肉并发出血的危险因素以及腺瘤性息肉的癌变特征分析

目的该研究目的是认识直径≥1.0 cm大肠息肉临床特点、内镜下治疗出血的危险因素及腺瘤性息肉癌变的特征分析。方法回顾该院2014年1月1日-2016年1月1日经内镜下切除的直径≥1.0 cm大肠息肉741例患者,共884枚息肉;分析患者的临床资料、内镜下特点、息肉切除出血危险因素及腺瘤性息肉癌变的特征。结果大肠息肉内镜下切除术术中、迟发性出血的单因素分析中,发现性别(P=0.017)、息肉部位(P=0.011)、息肉大小(P=0.004)、表面是否分叶(P=0.010)、内镜手术方式(P=0.029)在两组间差异有统计学意义;以息肉为观察单位,对患者临床资料和内镜下特点进行Logistic回归多因素分析时,发现性别(P=0.012,OR=2.671,95%CI=1.246~5.728)为独立危险因素,男性比女性更易发生出血;息肉部位为乙状结肠相对于直肠来讲为保护因素(P=0.011,OR=0.348,95%CI=0.154~0.786),息肉大小≥3.0 cm相对于息肉大小为1.0~1.9 cm来讲为危险因素(P=0.049,OR=2.530,95%CI=1.005~6.374)。大肠腺瘤性息肉癌变特征单因素分析时,发现表面是否分叶(P=0.001)、是否光滑(P=0.017)、山田分型(P=0.008)在两组间差异有统计学意义,进行Logistic回归分析多因素时,得出表面分叶(P=0.001,OR=6.556,95%CI=2.326~18.475)是腺瘤性息肉癌变的独立危险因素。结论内镜下治疗大肠息肉是一种安全的治疗方法 ;以息肉为观察单位,性别为大肠息肉内镜下切除术术中、迟发性出血的独立危险因素,男性比女性更易发生出血;息肉直径越大,发生出血的可能性越大;表面分叶的大肠腺瘤性息肉提示癌变的可能性大。

Portosplenomesenteric vein thrombosis in patients with early-stage severe acute pancreatitis

AIM To investigate the incidence and risk factors of portosplenomesenteric vein thrombosis(PSMVT) in the early stage of severe acute pancreatitis(SAP).METHODS Patients with SAP in a tertiary care setting from January 2014 to December 2016 were retrospectively reviewed. All contrast-enhanced computed tomography(CT) studies were reassessed and reviewed. Clinical outcome measures were compared between SAP patients with and without PSMVT in the early stage of the disease. Univariate and multivariate logistic regression analyses were sequentially performed to assess potential risk factors for the development of PSMVT in SAP patients. A receiver operating characteristic(ROC) curve was generated for the qualifying independent risk factors.RESULTS Twenty-five of the one hundred and forty(17.86%) SAP patients developed PSMVT 6.19 ± 2.43 d after acute pancreatitis(AP) onset. PSMVT was confirmed by contrast-enhanced CT. Multivariate stepwise logistic regression analyses showed that Balthazar's CT severity index(CTSI) scores [odds ratio(OR): 2.742; 95% confidence interval(CI): 1.664-4.519; P = 0.000], hypoalbuminemia(serum albumin level < 25 g/L)(OR: 32.573; 95%CI: 2.711-391.353; P = 0.006) and gastrointestinal wall thickening(OR: 4.367, 95%CI: 1.218-15.658; P = 0.024) were independent risk factors for PSMVT developed in patients with SAP. The area under the ROC curve for Balthazar's CTSI scores was 0.777(P = 0.000), the sensitivity was 52%, and the specificity was 93% at a cut-off value of 5.5.CONCLUSION High Balthazar's CTSI scores, hypoalbuminemia and gastrointestinal wall thickening are independent risk factors for PSMVT developed in the early stage of SAP.

Role of gut microbiota on intestinal barrier function in acute pancreatitis

Acute pancreatitis(AP)is a common gastrointestinal disorder.Approximately15%-20%of patients develop severe AP.Systemic inflammatory response syndrome and multiple organ dysfunction syndrome may be caused by the massive release of inflammatory cytokines in the early stage of severe AP,followed by intestinal dysfunction and pancreatic necrosis in the later stage.A study showed that 59%of AP patients had associated intestinal barrier injury,with increased intestinal mucosal permeability,leading to intestinal bacterial translocation,pancreatic tissue necrosis and infection,and the occurrence of multiple organ dysfunction syndrome.However,the real effect of the gut microbiota and its metabolites on intestinal barrier function in AP remains unclear.This review summarizes the alterations in the intestinal flora and its metabolites during AP development and progression to unveil the mechanism of gut failure in AP.

Helicobacter pylori infection and diabetes:Is it a myth or fact?

Helicobacter pylori (H. pylori) is one of the most common human bacterial pathogens, and infection causes a wide array of gastric disorders, including simple gastritis, peptic ulcers and gastric malignancies. Gastrointestinal inflammation caused by H. pylori can influence the absorption of glucose and lipids, which are also abnormal in diabetes mellitus. Type 2 diabetes mellitus (T2DM), formerly known as non-insulin-dependent diabetes mellitus or adult-onset diabetes, is a metabolic disorder that is characterized by high levels of blood glucose resulting from insulin resistance and relative insulin deficiency. It is an emerging pandemic and is rapidly becoming a serious threat to public health. Emerging data now indicate a strong relationship between H. pylori infection and the incidence of T2DM. The mechanisms underlying the pathogenesis of diabetes are complex, involving insulin resistance, chronic inflammation, insulin secretion deficiency as a result of pancreas &#x003b2;-cell dysfunction, glucotoxicity, and lipotoxicity. H. pylori infection is known to be involved in the pathogenesis of insulin resistance, and the growing awareness of its role in diabetes is important for the early detection of glucose dysregulation and prevention of T2DM in high-risk communities. This review probes the possible relationship between H. pylori and diabetes according to epidemiological surveys and discusses putative mechanisms underlying this correlation.

Th immune response induced by H pylori vaccine with chitosan as adjuvant and its relation to immune protection

AIM： To study the immunological protective effect of H pylori vaccine with chitosan as an adjuvant and its mechanism. METHODS： Female BALB/c mice were randomly divided into seven groups and orally immunized respectively with PBS, chitosan solution, chitosan particles, H pylori antigen, H pylori antigen plus cholera toxin （CT）, H pylori antigen plus chitosan solution, Hpylori antigen plus chitosan particles once a week for four weeks. Four weeks after the last immunization, the mice were challenged twice by alive Hpylori （1 × 10^9 CFU/mL） and sacrificed. Part of the gastric mucosa was embedded in paraffin, cut into sections and assayed with Giemsa staining. Part of the gastric mucosa was used to quantitatively culture Hpylori. EUSA was used to detect cytokine level in gastric mucosa and anti- Hpylori IgG1, IgG2a levels in serum. RESULTS： In the groups with chitosan as an adjuvant, immunological protection was achieved in 60% mice, which was significantly higher than in groups with H pylori antigen alone and without H pylori antigen （P 〈 0.05 or 0.001）. Before challenge, the level of IFN and IL-12 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in the control group and the group without adjuvant （P 〈 0.05 or 0.005）. After challenge, the level of IFN and IL-12 was significantly higher in the groups with adjuvant than in the groups without adjuvant and antigen （P 〈 0.05 or 0.001）. Before challenge, the level of IL-2 in gastric mucosa was not different among different groups. After challenge the level of IL-2 was significantly higher in the groups with adjuvant than in the control group （P 〈 0.05 or 0.001）. Before challenge, the level of IL-10 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in other groups without adjuvant （P 〈 0.05 or 0.01）. After challenge, the level of IL-10 was not different among different groups. Before challenge, the level of IL-4 in gastric mucosa was significantly higher in the groups with chitosan as an adjuvant than in other groups without adjuvant （P 〈 0.05）. After challenge, the level of IL-4 was significantly higher in the groups with chitosan particles as an adjuvant than in the group with CT as an adjuvant （P 〈 0.05）, and in the group with chitosan solution as an adjuvant, the level of IL-4 was significantly higher than that in control group, non-adjuvant group and the groups with CT （P 〈 0.05 or 0.001）. The ratio of anti- Hpylori IgG2a/ IgG1 in serum was significantly lower in the groups with chitosan as an adjuvant than in the groups with CT as an adjuvant or without adjuvant （P 〈 0.01）. CONCLUSION： H pylori vaccine with chitosan as an adjuvant can protect against H pylori infection and induce both Thl and Th2 type immune response.

Furazolidone-based triple and quadruple eradication therapy for Helicobacter pylori infection

AIM: To evaluate the efficacy of furazolidone-based triple and quadruple therapy in eradicating Helicobacter pylori (H. pylori) in a multi-center randomized controlled trial.

Severity of acute gastrointestinal injury grade is a good predictor of mortality in critically ill patients with acute pancreatitis

BACKGROUND Gastrointestinal(GI)dysfunction is a common and important complication of acute pancreatitis(AP),especially in patients with severe AP.Despite this,there is no consensus means of obtaining a precise assessment of GI function.AIM To determine the association between acute gastrointestinal injury(AGI)grade and clinical outcomes in critically ill patients with AP.METHODS Patients with AP admitted to our pancreatic intensive care unit from May 2017 to May 2019 were enrolled.GI function was assessed according to the AGI grade proposed by the European Society of Intensive Care Medicine in 2012,which is mainly based on GI symptoms,intra-abdominal pressure,and feeding intolerance in the first week of admission to the intensive care unit.Multivariate logistic regression analysis was performed to assess the association between AGI grade and clinical outcomes in critically ill patients with AP.RESULTS Among the 286 patients included,the distribution of patients with various AGI grades was 34.62%with grade I,22.03%with grade II,32.52%with grade III,and 10.84%with grade IV.The distribution of mortality was 0%among those with grade I,6.35%among those with grade II,30.11%among those with grade III,and 61.29%among those with grade IV,and AGI grade was positively correlated with mortality(χ2=31.511,P<0.0001).Multivariate logistic regression analysis showed that age,serum calcium level,AGI grade,persistent renal failure,and persistent circulatory failure were independently associated with mortality.Compared with the Acute Physiology and Chronic Health Evaluation II score(area under the curve:0.739 vs 0.854;P<0.05)and Ranson score(area under the curve:0.72 vs 0.854;P<0.01),the AGI grade was more useful for predicting mortality.CONCLUSION AGI grade is useful for identifying the severity of GI dysfunction and can be used as a predictor of mortality in critically ill patients with AP.

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).Moreover,Ku70/80 expression was negatively correlated with both clinical stage(r=-0.189,P<0.05)and H.pylori colonization(r=-0.168,P<0.05).CONCLUSION:Overall,this research demonstrated that enhanced DNA-PKcs and Ku 70/80 expression may be closely associated with gastric carcinoma.

Is NEDD4-1 a negative regulator of phosphatase and tensin homolog in gastric carcinogenesis?

The expression of phosphatase and tensin homolog (PTEN ), a tumor suppressor gene, is frequently downregulated in gastric carcinomas due to mutation, loss of heterozygosity, and promoter hypermethylation. However, it is unknown if additional mechanisms may account for the down-regulation of PTEN expression. While neuronal precursor cell-expressed developmentally down-regulated 4-1 (NEDD4-1) is believed to be a potential dual regulator of PTEN, there are conflicting reports regarding their interaction. To gain further insight into the role of NEDD4-1 and its association with PTEN in gastric carcinoma development, we measured the protein expression of NEDD4-1 and PTEN in gastric mucosae with various pathological lesions and found that NEDD4-1 increased from normal gastric mucosa to intestinal metaplasia and decreased from dysplasia to gastric carcinoma. These changes did not correlate with PTEN expression changes during gastric carcinogenesis. Moreover, we found similar results in protein levels in the primary tumors and adjacent non-tumorous tissues. These results differ from a previous report showing that expression of NEDD4-1 is up-regulated in gastric carcinomas, and show a more complex pattern of NEDD4-1 gene expression during gastric carcinogenesis.

Helicobacter pylori infection in Mongolian gerbils does not initiate hematological diseases

AIM: To investigate whether Helicobacter pylori (H. pylori) infection contributes to idiopathic thrombocytopenic purpura (ITP) or iron-deficiency anemia (IDA) onset in gerbils.

Pancreatic necrosis and severity are independent risk factors for pancreatic endocrine insufficiency after acute pancreatitis: A long-term follow-up study

BACKGROUND Pancreatic endocrine insufficiency after acute pancreatitis(AP) has drawn increasing attention in recent years.AIM To assess the impact of risk factors on the development of pancreatic endocrine insufficiency after AP.METHODS This retrospective observational long-term follow-up study was conducted in a tertiary hospital. Endocrine function was evaluated by the oral glucose tolerance test. The data, including age, sex, body mass index, APACHE II score, history of smoking and drinking, organ failure, pancreatic necrosis, debridement of necrosis(minimally invasive and/or open surgery), and time interval, were collected from the record database.RESULTS A total of 361 patients were included in the study from January 1, 2012 to December 30, 2018. A total of 150(41.6%) patients were diagnosed with dysglycemia(including diabetes mellitus and impaired glucose tolerance), while211(58.4%) patients had normal endocrine function. The time intervals(mo) of the above two groups were 18.73 ± 19.10 mo and 31.53 ± 27.27 mo, respectively(P= 0.001). The morbidity rates of pancreatic endocrine insufficiency were 46.7%,28.0%, and 25.3%, respectively, in the groups with different follow-up times. The risk factors for pancreatic endocrine insufficiency after AP were severity(odds ratio [OR] = 3.489;95% confidence interval [CI]: 1.501-8.111;P = 0.004) and pancreatic necrosis(OR = 4.152;95%CI: 2.580-6.684;P = 0.001).CONCLUSION Pancreatic necrosis and severity are independent risk factors for pancreatic endocrine insufficiency after AP. The area of pancreatic necrosis can affect pancreatic endocrine function.

Recent progress in Helicobacter pylori treatment

The main challenge in the field of Helicobacter pylori(H.pylori)infection is antibiotic resistance,which influences the efficacy of eradication regimens.Bismuth-containing quadruple therapy has been confirmed as an effective regimen for eradicating H.pylori,especially in strains with antibiotic resistance.High-dose proton-pump inhibitor-amoxicillin dual therapy could decrease the use of unnecessary antibiotics,which is a promising alternative approach.Adjuvant therapy(specific probiotic or vitamin)also showed good results,although more evidence is needed.Novel anti-H.pylori drugs are needed,and the establishment of the H.pylori database is an effective way to acknowledge the real-time information of H.pylori management.This review provides the recent progress of H.pylori treatment,and further studies are needed to address the role of different regimens in improving H.pylori eradication rate,especially in strains with antibiotics resistance.