Helicobacter pylori infection induces POU5F1 upregulation and SPP1 activation to promote chemoresistance and T cell inactivation in gastric cancer cells.

Infection with Helicobacter pylori (H. pylori or Hp) is associated with an increased susceptibility to gastric diseases, notably gastric cancer (GC). This study investigates the impact of Hp infection on chemoresistance and immune activity in GC cells. Hp infection in AGS and MKN-74 cells promoted proliferation, migration and invasion, apoptosis resistance, and tumorigenic activity of cells under cisplatin (DDP) plus gemcitabine (GEM) treatment. Additionally, it dampened activity of the co-cultured CD8+ T cells. Hp infection increased POU class 5 homeobox 1 (POU5F1) level, which further activated secreted phosphoprotein 1 (SPP1) transcription to increase its expression. Silencing of either SPP1 or POU5F1 enhanced the GEM sensitivity in GC cells, and it increased the populations of CD8+ T cells and the secretion of immune-active cytokines both in vitro and in xenograft tumors in immunocompetent mice. However, the effects of POU5F1 silencing were counteracted by SPP1 overexpression. Furthermore, the POU5F1/SPP1 axis activated the PI3K/AKT signaling pathway. This study demonstrates that Hp infection induces POU5F1 upregulation and SPP1 activation, leading to increased DDP/GEM resistance and T cell inactivation in GC cells.

Autoimmune pancreatitis type 2 (idiopathic duct-centric pancreatitis): A comprehensive review.

Autoimmune pancreatitis (AIP) is an uncommon fibro-inflammatory disorder precipitated by autoimmune/inflammatory reactions. Currently, there are two clinical subtypes of AIP (type 1 [AIP-1] and type 2 [AIP-2]) that correspond to two histologic descriptors (lymphoplasmacytic sclerosing pancreatitis and idiopathic duct-centric pancreatitis, respectively). While our understanding of AIP-1 has evolved considerably over the years, little is known about AIP-2 due to its rarity, often leading to misdiagnosis, delayed treatment, and even unnecessary surgical resection. Compared to AIP-1, AIP-2 exhibits distinct clinical and histologic features. Because AIP-2 is a pancreas-restricted disease without a specific serum marker, the evaluation of histologic features (e.g., granulocytic epithelial lesions) is essential for an accurate diagnosis. Patients with AIP-2 respond well to glucocorticoids, with anti-tumor necrosis factor-alpha antibodies as a promising alternative therapy. The prognosis of AIP-2 is generally favorable and relapse is uncommon. Here, we provide an overview of our current knowledge on the clinical features, diagnosis, therapeutic regimens, prognosis, and putative mechanisms underlying AIP-2. Notably, the diagnostic differentiation between AIP-2, especially the mass-forming/focal type, and pancreatic cancer is important, but challenging. In this regard, endoscopic ultrasound-guided core biopsy has a key role, but novel diagnostic markers and modalities are clearly needed.

MicroRNA-1-3p enhances osteoblast differentiation of MC3T3-E1 cells by interacting with hypoxia-inducible factor 1 α inhibitor (HIF1AN).

Studies have proved that miRNAs participate in the regulation of osteoblast differentiation (OD), and abnormal expression of miRNAs is related with various states of OD. In this study, we investigated the role of miRNA-1-3p in OD using MC3T3-E1 cells. BMP2 is used to induce OD of MC3T3-E1 cells. MiRNA-1-3p mimics or miRNA-1-3p inhibitor was transfected to MC3T3-E1 cells with BMP2. The expression levels of miRNA-1-3p were determined by qRT-PCR. The expression of Runx2, OSX, OPN, and OCN was detected by Western blotting. ALP assay was performed to measure alkaline phosphatase activity. Calcium nodules were evaluated by alizarin red staining. Over-expression of hypoxia-inducible factor 1-alpha inhibitor (HIF1AN) was performed and miRNA-1-3p rescue experiments were carried out. Over-expression of miRNA-1-3p promoted osteogenic differentiations and calcifications, as demonstrated by increased ALP, calcification and osteogenic markers. Knock-down of miRNA-1-3p generated the opposite results. HIF1AN was identified to be directly targeted by miRNA-1-3p. Over-expression of HIF1AN suppressed OD and calcifications, and miRNA-1-3p reversed the effect. Our results demonstrated that miRNA-1-3p could enhance OD of MC3T3-E1 cells through interacting with HIF1AN, which might be employed as therapeutic applications for bone formation and regeneration.

Polyphyllin VII attenuated RANKL-induced osteoclast differentiation via inhibiting of TRAF6/c-Src/PI3K pathway and ROS production.

BACKGROUND: Osteoporosis is a worldwide severe bone disease. This study aimed to evaluate the effect of polyphyllin VII on the genesis of osteoclasts from bone marrow macrophages (BMMs) and its potentiality as a therapeutic drug for osteoporosis. METHODS: BMMs were induced to differentiate into osteoclasts by RANKL and M-CSF. The cells were then treated with various concentrations of polyphyllin VII. Intracellular reactive oxygen species (ROS) measurement assay, resorption pit formation assay, tartrate-resistant acid phosphatase (TRAP) staining and TRAP activity assessment, cell viability assay, active GTPase pull-down assay, immunofluorescent staining, immunoblotting, and RT-PCR were performed. RESULTS: RANKL + M-CSF significantly increased TRAP activity, number of osteoclasts, number and area of lacunae, intracellular content of ROS, protein levels of Nox1, TRAF6, c-Src and p-PI3K, as well as the content of activated GTP-Rac1, which were significantly blocked by polyphyllin VII in a concentration-dependent manner. CONCLUSION: These findings suggested that polyphyllin VII inhibited differentiation of BMMs into osteoclasts through suppressing ROS synthesis, which was modulated by TRAF6-cSrc-PI3k signal transduction pathway including GTP-Rac1 and Nox1. Polyphyllin VII could be a therapeutic drug for osteoporosis.

Probiotic effect on Helicobacter pylori attachment and inhibition of inflammation in human gastric epithelial cells.

Helicobacter pylori (H. pylori) is a major cause of chronic gastritis, gastric ulcers and gastric cancer. Recent studies have identified that probiotics are beneficial to human health due, in part, to their anti-H. pylori activities. Therefore, the present study investigated the antagonistic and local immunoregulatory activities of seven commercial probiotic strains and explored their mechanisms of actions. The human gastric epithelial cell line-1 (GES-1) was used to assess the effects of probiotics on the adhesion ability of H. pylori. GES-1 cells were infected with H. pylori plus lipopolysaccharide (HP-LPS) or the drug-resistant H. pylori strain (HP021) in the presence or absence of live probiotics. Following this, the growth rate and the adhesion ability of GES-1 cells were detected using MTT and urease activity assay. Toll-like receptor 4 (TLR4), NFKB inhibitor-α (IκBα) and nuclear factor (NF)-κB levels were measured by western blot analysis. The amount of interleukin (IL)-8 in the cell culture medium was determined by ELISA. Amongst the seven probiotic strains studied, live Lactobacillus acidophilus (L. acidophilus) and Lactobacillus bulgaricus (L. bulgaricus) inhibited H. pylori adherence to GES-1 cells most significantly. L. bulgaricus inhibited IL-8 production by GES-1 cells through modulation of the TLR4/IκBα/NF-κB pathway. Therefore, the present results suggested that consumption of food containing L. acidophilus and L. bulgaricus may be used as an adjuvant therapy for H. pylori-associated gastritis.

MicroRNA­100­5p inhibits osteoclastogenesis and bone resorption by regulating fibroblast growth factor 21.

MicroRNAs (miRNAs/miRs) are post­transcriptional regulators that serve important roles in osteoclastogenesis and bone metabolism; however, the roles of miRNAs have not been completely clarified. The present study aimed to investigate the effects of miR­100­5p on the mechanism of liver­bone endocrine metabolism. A miRNA microarray analysis was conducted to evaluate the miRNA expression profile during receptor activator of nuclear factor­κB ligand­stimulated osteoclastogenesis. Hematoxylin and eosin and tartrate­resistant acid phosphatase staining were performed to analyze the trabecular bone microstructure and osteoclast differentiation. The mRNA and protein expression levels were assessed by reverse transcription­quantitative polymerase chain reaction and western blotting, respectively. The results revealed that in vitro osteoclast differentiation and in vivo bone resorption were suppressed by miR­100­5p overexpression. In vivo, a decrease in miR­100­5p and an increase in FGF21 were simultaneously observed in mice following ovariectomy (OVX). Bioinformatics analysis and experimental data confirmed that FGF21 was a direct target of miR­100­5p. Conversely, augmentation of miR­100­5p using a specific agomir in OVX­operated mice decreased the levels of FGF21 in the serum and liver, and prevented osteoclastogenesis and bone loss. The present study revealed that FGF21 may be a signal molecule associated with the mechanism of liver­bone endocrine metabolism and may be targeted by miR­100­5p. In addition, miR­100­5p may serve an important role in protecting against OVX­induced osteoporosis.

What Roles Do Probiotics Play in the Eradication of Helicobacter pylori? Current Knowledge and Ongoing Research.

With the rising global prevalence of antibiotic resistance, the eradication rate of Helicobacter pylori (HP) is continuing to decrease. Probiotics are beneficial to human health and may be an adjunct therapy to increase the eradication rate of HP, lower treatment-associated side effects, and reduce HP-associated gastric inflammation. However, inconsistent test results have prevented conclusions about the therapeutic prowess of probiotics for HP. The mechanisms of actions of probiotics include the production of substances that inhibit or kill HP or compete with HP for the adhesion site on gastric epithelial cells. Probiotics can also reduce the release of inflammatory factors by regulating the local immune response of the host. We searched the available literature for full-length articles focusing on the role of probiotics in HP management. This review presents the latest advances in this area.

Can eradication rate of gastric Helicobacter pylori be improved by killing oral Helicobacter pylori?

AIM: To evaluate the influence of oral Helicobacter pylori (H. pylori) on the success of eradication therapy against gastric H. pylori. METHODS: A total of 391 patients with dyspepsia were examined for H. pylori using the saliva H. pylori antigen test (HPS), ¹³C-urea breath test (UBT), gastroscopy, and gastric mucosal histopathological detection. Another 40 volunteers without discomfort were subjected to HPS and ¹³C-UBT, and served as the control group. The 233 patients who were (¹³C-UBT+ were enrolled in this study and divided into 4 groups. Patients who were HPS- and ¹³C-UBT+ (n = 53) received triple therapy alone. Those who were both HPS+ and ¹³C-UBT+ (n = 180) were randomly divided into 3 groups: (1) the O+G+t group which received triple therapy alone (n = 53); (2) the O+G+tm group which received both triple therapy and mouthrinse treatment (n = 65); and (3) the O+G+tmp group which received triple therapy, mouthrinse, and periodontal treatment (n = 62). The HPS and ¹³C-UBT were continued for 4 wk after completion of treatment, and the eradication rate of gastric H. pylori and the prevalence of oral H. pylori in the 4 groups were then compared. RESULTS: The eradication rates of gastric H. pylori in the O-G+t group, the O+G+tm group, and the O+G+tmp group were 93.3%, 90.0%, and 94.7% respectively; all of these rates were higher than that of the O+G+t group (78.4%) [O-G+t group vs O+G+t group (P = 0.039); O+G+tm group vs O+G+t group (P = 0.092); O+G+tmp group vs O+G+t group (P = 0.012); O+G+tm group vs O-G+t group (P = 0.546); O+G+tmp group vs O-G+t group (P = 0.765); O+G+tm group vs O+G+tmp group (P = 0.924)]. The eradication of gastric H. pylori was significantly improved using the combination of triple therapy, mouthrinse, and periodontal treatment. The eradication rates of gastric H. pylori in the peptic ulcer group, chronic atrophic gastritis group and control group were higher than in the duodenitis group and the superficial gastritis group. The prevalence rates of oral H. pylori in the O-G+t group, O+G+t group, O+G+tm group and O+G+tmp group following treatment were 0%, 76.5%, 53.3%, and 50.9%, respectively [O-G+t group vs O+G+t group (P < 0.0001); O+G+tm group vs O+G+t group (P = 0.011); O+G+tmp group vs O+G+t group (P = 0.006); O+G+tm group vs O-G+t group (P < 0.0001); O+G+tmp group vs O-G+t group (P < 0.0001); O+G+tm group vs the O+G+tmp group (P = 0.790)]. Both mouthrinse and periodontal treatment significantly reduced the prevalence of oral H. pylori. CONCLUSION: Mouthrinse treatment alone or combined with periodontal treatment can, to some extent, reduce the prevalence of oral H. pylori and improve the eradication rate of gastric H. pylori.