Quercetin alleviates hyperoxia-induced bronchopulmonary dysplasia by inhibiting ferroptosis through the MAPK/PTGS2 pathway: Insights from network pharmacology, molecular docking, and experimental evaluations.

Quercetin, a bioactive natural compound renowned for its potent anti-inflammatory, antioxidant, and antiviral properties, has exhibited therapeutic potential in various diseases. Given that bronchopulmonary dysplasia (BPD) development is closely linked to inflammation and oxidative stress, and quercetin, a robust antioxidant known to activate NRF2 and influence the ferroptosis pathway, offers promise for a wide range of age groups. Nonetheless, the specific role of quercetin in BPD remains largely unexplored. This study aims to uncover the target role of quercetin in BPD through a combination of network pharmacology, molecular docking, computer analyses, and experimental evaluations.

BCL6 attenuates hyperoxia-induced lung injury by inhibiting NLRP3-mediated inflammation in fetal mouse.

BACKGROUND: The transcriptional repressor B-cell lymphoma 6 (BCL6) has been reported to inhibit inflammation. So far, experimental evidence for the role of BCL6 in bronchopulmonary dysplasia (BPD) is lacking. Our study investigated the roles of BCL6 in the progression of BPD and its downstream mechanisms. METHODS: Hyperoxia or lipopolysaccharide (LPS) was used to mimic the BPD mouse model. To investigate the effects of BCL6 on BPD, recombination adeno-associated virus serotype 9 expressing BCL6 (rAAV9-BCL6) and BCL6 inhibitor FX1 were administered in mice. The pulmonary pathological changes, inflammatory chemokines and NLRP3-related protein were observed. Meanwhile, BCL6 overexpression plasmid was used in human pulmonary microvascular endothelial cells (HPMECs). Cell proliferation, apoptosis, and NLRP3-related protein were detected. RESULTS: Either hyperoxia or LPS suppressed pulmonary BCL6 mRNA expression. rAAV9-BCL6 administration significantly inhibited hyperoxia-induced NLRP3 upregulation and inflammation, attenuated alveolar simplification and dysregulated angiogenesis in BPD mice, which were characterized by decreased mean linear intercept, increased radical alveolar count and alveoli numbers, and the upregulated CD31 expression. Meanwhile, BCL6 overexpression promoted proliferation and angiogenesis, inhibited apoptosis and inflammation in hyperoxia-stimulated HPMECs. Moreover, administration of BCL6 inhibitor FX1 arrested growth and development. FX1-treated BPD mice exhibited exacerbation of alveolar pathological changes and pulmonary vessel permeability, with upregulated mRNA levels of pro-inflammatory cytokines and pro-fibrogenic factors. Furthermore, both rAAV9-BCL6 and FX1 administration exerted a long-lasting effect on hyperoxia-induced lung injury (≥4 wk). CONCLUSIONS: BCL6 inhibits NLRP3-mediated inflammation, attenuates alveolar simplification and dysregulated pulmonary vessel development in hyperoxia-induced BPD mice. Hence, BCL6 may be a target in treating BPD and neonatal diseases.

Exposure to irregular microplastic shed from baby bottles activates the ROS/NLRP3/Caspase-1 signaling pathway, causing intestinal inflammation.

Irregularly shaped microplastics (MPs) released from infant feeding bottles (PP-IFBs) may exhibit increased cytotoxicity, in contrast to the commonly studied spherical MPs. This study presents an initial analysis of the thermal-oxidative aging process of plastic shedding from feeding bottles, and investigates the inflammatory response induced by these atypical MPs in human intestinal cells (Caco-2). The PP-IFBs' surface displayed non-uniform white patches and increased roughness, revealing substantial structural alteration and shedding, especially during actions such as shaking, boiling water disinfection, and microwave heating. FT-IR and 2D-COS analyses revealed that oxygen targeted the C-H and C-C bonds of polypropylene molecular chain, producing RO· and ·OH, thereby hastening polypropylene degradation. When human intestinal cells were exposed to MPs from PP-IFBs, oxidative stress was triggered, resulting in lowered glutathione levels, augmented reactive oxygen species (ROS), and heightened lipid peroxidation. Elevated levels of pro-inflammatory cytokines (IL-6 and TNFα) signified an active inflammatory process. The inflammatory response was notably more intense when exposed to MPs released through boiling water disinfection and microwave heating treatments, primarily due to the larger quantity of MPs released and their higher proportion of smaller particles. Furthermore, the NLRP3 inflammasome was identified as critical in initiating this inflammatory chain reaction due to the mitochondrial ROS surge caused by MPs exposure. This was further validated by inhibitor studies, emphasizing the role of the ROS/NLRP3/Caspase-1/IL-1ß signaling pathway in in promoting intestinal inflammation. Therefore, swift actions are recommended to protect infants against the potential health effects of MPs exposure.

Lactobacillus reuteri in digestive system diseases: focus on clinical trials and mechanisms.

Objectives: Digestive system diseases have evolved into a growing global burden without sufficient therapeutic measures. Lactobacillus reuteri (L. reuteri) is considered as a new potential economical therapy for its probiotic effects in the gastrointestinal system. We have provided an overview of the researches supporting various L. reuteri strains' application in treating common digestive system diseases, including infantile colic, diarrhea, constipation, functional abdominal pain, Helicobacter pylori infection, inflammatory bowel disease, diverticulitis, colorectal cancer and liver diseases. Methods: The summarized literature in this review was derived from databases including PubMed, Web of Science, and Google Scholar. Results: The therapeutic effects of L. reuteri in digestive system diseases may depend on various direct and indirect mechanisms, including metabolite production as well as modulation of the intestinal microbiome, preservation of the gut barrier function, and regulation of the host immune system. These actions are largely strain-specific and depend on the activation or inhibition of various certain signal pathways. It is well evidenced that L. reuteri can be effective both as a prophylactic measure and as a preferred therapy for infantile colic, and it can also be recommended as an adjuvant strategy to diarrhea, constipation, Helicobacter pylori infection in therapeutic settings. While preclinical studies have shown the probiotic potential of L. reuteri in the management of functional abdominal pain, inflammatory bowel disease, diverticulitis, colorectal cancer and liver diseases, its application in these disease settings still needs further study. Conclusion: This review focuses on the probiotic effects of L. reuteri on gut homeostasis via certain signaling pathways, and emphasizes the importance of these probiotics as a prospective treatment against several digestive system diseases.

Molecular mechanisms of cell death in bronchopulmonary dysplasia.

Bronchopulmonary dysplasia (BPD) in neonates is the most common pulmonary disease that causes neonatal mortality, has complex pathogenesis, and lacks effective treatment. It is associated with chronic obstructive pulmonary disease, pulmonary hypertension, and right ventricular hypertrophy. The occurrence and development of BPD involve various factors, of which premature birth is the most crucial reason for BPD. Under the premise of abnormal lung structure and functional product, newborns are susceptible to damage to oxides, free radicals, hypoxia, infections and so on. The most influential is oxidative stress, which induces cell death in different ways when the oxidative stress balance in the body is disrupted. Increasing evidence has shown that programmed cell death (PCD), including apoptosis, necrosis, autophagy, and ferroptosis, plays a significant role in the molecular and biological mechanisms of BPD and the further development of the disease. Understanding the mode of PCD and its signaling pathways can provide new therapeutic approaches and targets for the clinical treatment of BPD. This review elucidates the mechanism of BPD, focusing on the multiple types of PCD in BPD and their molecular mechanisms, which are mainly based on experimental results obtained in rodents.

Effects of Antioxidants in Human Milk on Bronchopulmonary Dysplasia Prevention and Treatment: A Review.

Bronchopulmonary dysplasia (BPD) is a severe chronic lung illness that affects neonates, particularly premature infants. It has far-reaching consequences for infant health and their families due to intractable short- and long-term repercussions. Premature infant survival and long-term quality of life are severely harmed by BPD, which is characterized by alveolarization arrest and hypoplasia of pulmonary microvascular cells. BPD can be caused by various factors, with oxidative stress (OS) being the most common. Premature infants frequently require breathing support, which results in a hyperoxic environment in the developing lung and obstructs lung growth. OS can damage the lungs of infants by inducing cell death, inhibiting alveolarization, inducing inflammation, and impairing pulmonary angiogenesis. Therefore, antioxidant therapy for BPD relieves OS and lung injury in preterm newborns. Many antioxidants have been found in human milk, including superoxide dismutase, glutathione peroxidase, glutathione, vitamins, melatonin, short-chain fatty acids, and phytochemicals. Human milk oligosaccharides, milk fat globule membrane, and lactoferrin, all unique to human milk, also have antioxidant properties. Hence, human milk may help prevent OS injury and improve BPD prognosis in premature infants. In this review, we explored the role of OS in the pathophysiology of BPD and related signaling pathways. Furthermore, we examined antioxidants in human milk and how they could play a role in BPD to understand whether human milk could prevent and treat BPD.

Nonenhanced MRI-based radiomics model for preoperative prediction of nonperfused volume ratio for high-intensity focused ultrasound ablation of uterine leiomyomas.

OBJECTIVES: To develop and assess nonenhanced MRI-based radiomics model for the preoperative prediction of nonperfused volume (NPV) ratio of uterine leiomyomas after high-intensity focused ultrasound (HIFU) treatment. METHODS: Two hundred and five patients with uterine leiomyomas treated by HIFU were enrolled and allocated to training (N =164) and testing cohorts (N = 41). Pyradiomics was used to extract radiomics features from T2-weighted images and apparent diffusion coefficient (ADC) map generated from diffusion-weighted imaging (DWI). The clinico-radiological model, radiomics model, and radiomics-clinical model which combined the selected radiomics features and clinical parameters were used to predict technical outcomes determined by NPV ratios where three classification groups were created (NPV ratio ≤ 50%, 50-80% or ≥ 80%). The receiver operating characteristic (ROC) curve, area under the curve (AUC), and calibration and decision curve analyses were performed to illustrate the prediction performance and clinical usefulness of model in the training and testing cohorts. RESULTS: The multi-parametric MRI-based radiomics model outperformed T2-weighted imaging (T2WI)-based radiomics model, which achieved an average AUC of 0.769 (95% confidence interval [CI], 0.701-0.842), and showed satisfactory prediction performance for NPV ratio classification. The radiomics-clinical model demonstrated best prediction performance for HIFU treatment outcome, with an average AUC of 0.802 (95% CI, 0.796-0.850) and an accuracy of 0.762 (95% CI, 0.698-0.815) in the testing cohort, compared to the clinico-radiological and radiomics models. The decision curve also indicated favorable clinical usefulness of the radiomics-clinical model. CONCLUSIONS: Nonenhanced MRI-based radiomics has potential in the preoperative prediction of NPV ratio for HIFU ablation of uterine leiomyomas.

Prediction of Clinical Outcome for High-Intensity Focused Ultrasound Ablation of Uterine Leiomyomas Using Multiparametric MRI Radiomics-Based Machine Leaning Model.

OBJECTIVES: This study sought to develop a multiparametric MRI radiomics-based machine learning model for the preoperative prediction of clinical success for high-intensity-focused ultrasound (HIFU) ablation of uterine leiomyomas. METHODS: One hundred and thirty patients who received HIFU ablation therapy for uterine leiomyomas were enrolled in this retrospective study. Radiomics features were extracted from T2-weighted (T2WI) image and ADC map derived from diffusion-weighted imaging (DWI). Three feature selection algorithms including least absolute shrinkage and selection operator (LASSO), recursive feature elimination (RFE), and ReliefF algorithm were used to select radiomics features, respectively, which were fed into four machine learning classifiers including k-nearest neighbors (KNN), logistic regression (LR), random forest (RF), and support vector machine (SVM) for the construction of outcome prediction models before HIFU treatment. The performance, predication ability, and clinical usefulness of these models were verified and evaluated using receiver operating characteristics (ROC), calibration, and decision curve analyses. RESULTS: The radiomics analysis provided an effective preoperative prediction for HIFU ablation of uterine leiomyomas. Using SVM with ReliefF algorithm, the multiparametric MRI radiomics model showed the favorable performance with average accuracy of 0.849, sensitivity of 0.814, specificity of 0.896, positive predictive value (PPV) of 0.903, negative predictive value (NPV) of 0.823, and the area under the ROC curve (AUC) of 0.887 (95% CI = 0.848-0.939) in fivefold cross-validation, followed by RF with ReliefF. Calibration and decision curve analyses confirmed the potential of model in predication ability and clinical usefulness. CONCLUSIONS: The radiomics-based machine learning model can predict preoperatively HIFU ablation response for the patients with uterine leiomyomas and contribute to determining individual treatment strategies.

A Prediction of NPVR ≥ 80% of Ultrasound-Guided High-Intensity Focused Ultrasound Ablation for Uterine Fibroids.

Objective: To evaluate factors in predicting the treatment outcome of ultrasound-guided high-intensity focused ultrasound (USgHIFU) ablation for uterine fibroids with a non-perfused volume ratio (NPVR) of at least 80%. Methods: One thousand patients with uterine fibroids who received USgHIFU were enrolled. Thirty-two independent variables of four dimensions of data set, including general information of patients, clinical symptoms, laboratory tests, and fibroid imaging characteristics, were used to investigate the potential predictors of the NPVR of at least 80% by multivariate logistic regression. NPVR was the gold standard for evaluating the efficiency of HIFU ablation, and a NPVR of at least 80% was considered sufficient ablation, while partial ablation was defined as having an NPVR of <80%. Results: Out of 1,000 fibroids, 758 obtained sufficient ablation and 242 obtained partial ablation, and the median NPVR was 88.3% (interquartile range: 80.3-94.8%). The probability of NPVR reaching 80% fibroids with a signal intensity of T2WI of hypointense, isointense, and hyperintense was 86.4, 76.5, and 62.6%, respectively; fibroids with an enhancement type of T1WI of slight, irregular, and regular was 81.5, 73.6, and 63.7%, respectively; and fibroids with uterine anteroposterior of 30-130 mm was 57.7-78.3%, respectively. In patients with a platelet count of 50 × 109/L-550 × 109/L, the probability of NPVR reaching 80% is from 53.4 to 80.1%, respectively. Conclusions: In predicting NPVR ≥ 80%, the signal intensity on T2WI was the most important factor affecting ablative efficiency, followed by enhancement type on T1WI, uterine anteroposterior, and platelet count.

Sodium Propionate Enhances Nrf2-Mediated Protective Defense Against Oxidative Stress and Inflammation in Lipopolysaccharide-Induced Neonatal Mice.

BACKGROUND: Alveolar arrest and the impaired angiogenesis caused by chronic inflammation and oxidative stress are two main factors in bronchopulmonary dysplasia (BPD). Short-chain fatty acids (SCFAs), especially propionate, possess anti-oxidant and anti-inflammatory effects. The present study was designed to examine the roles of sodium propionate (SP) on lipopolysaccharide (LPS)-challenged BPD and its potential mechanisms. METHODS: WT, Nrf2-/- mice and pulmonary microvascular endothelial cells (HPMECs) were used in this study. LPS was performed to mimic BPD model both in vivo and vitro. Lung histopathology, inflammation and oxidative stress-related mRNA expressions in lungs involved in BPD pathogenesis were investigated. In addition, cell viability and angiogenesis were also tested. RESULTS: The increased nuclear factor erythroid 2-related factor (Nrf2) and decreased Kelch-like ECH-associated protein-1 (Keap-1) expressions were observed after SP treatment in the LPS-induced neonatal mouse model of BPD. In LPS-induced wild-type but not Nrf2-/- neonatal mice, SP reduced pulmonary inflammation and oxidative stress and exhibited obvious pathological alterations of the alveoli. Moreover, in LPS-evoked HPMECs, SP accelerated Nrf2 nuclear translocation presented and exhibited cytoprotective and pro-angiogenesis effects. In addition, SP diminished the LPS-induced inflammatory response by blocking the activation of nuclear factor-kappa B pathway. Moreover, pretreatment with ML385, an Nrf2 specific inhibitor, offsets the beneficial effects of SP on inflammation, oxidative stress and angiogenesis in LPS-evoked HPMECs. CONCLUSION: SP protects against LPS-induced lung alveolar simplification and abnormal angiogenesis in neonatal mice and HPMECs in an Nrf2-dependent manner.

Comparison of Dose and Effectiveness of a Single-Session Ultrasound-Guided High-Intensity Focused Ultrasound Ablation of Uterine Fibroids With Different Sizes.

PURPOSE: This study aimed to compare the dose and effectiveness of ultrasound-guided high-intensity focused ultrasound (USgHIFU) ablation of uterine fibroids with different sizes and explore the effect of uterine fibroid size on dose, which provided dose evaluation for clinicians in accordance with the size of uterine fibroids. MATERIALS AND METHODS: A total of 1,000 patients with symptomatic uterine fibroids who received a single-session USgHIFU treatment were enrolled in this study. The size of fibroids was divided into seven groups: 3-4 cm, 4-5 cm, 5-6 cm, 6-7 cm, 7-8 cm, 8-9 cm, and 9-11 cm. The dose was expressed on the basis of the energy efficiency factor (EEF) as the energy required for ablation per unit volume of tissue, and the non-perfused volume ratio (NPVR) was used to assess the effect of HIFU ablation. RESULTS: The median NPVR of 88.3% (IQR: 80.3%-94.8%) was obtained, and no significant difference was observed among the seven groups. The classification of T2-weighted image signal intensity fibroids in the 4-5 cm group was compared with that in the 6-7 cm and 8-9 cm groups, and the difference was significant (p < 0.05). However, the proportion of T2WI hyperintense signal fibroids had no significant difference among the seven groups (p > 0.05). The median EEF was 3.88 J/mm3, and a significant difference was observed among the seven groups of EEF (p < 0.05). The EEF of groups with a fibroid size less than 6 cm was more than double the EEF of groups with a fibroid size above 6 cm. In addition, the EEF of groups with a fibroid size of 4-5 cm and 3-4 cm was 3-4 times higher than those with a fibroid size above 7 cm (p < 0.05). CONCLUSIONS: A single-session HIFU ablation for uterine fibroids of 3-11 cm can obtain an NPVR of more than 80%. The EEF decreased with the increase of the size of uterine fibroids. A fibroid size of 6.5 cm was considered as a clinical meaningful point affecting EEF.

Sodium Propionate Attenuates the Lipopolysaccharide-Induced Epithelial-Mesenchymal Transition via the PI3K/Akt/mTOR Signaling Pathway.

Short-chain fatty acids (SCFAs), especially propionate, originate from the fermentation of dietary fiber in the gut and play a key role in inhibiting pulmonary inflammation. Chronic inflammation may induce an epithelial-mesenchymal transition (EMT) in alveolar epithelial cells and result in fibrotic disorders. This study was designed to investigate the beneficial effect of sodium propionate (SP) on lipopolysaccharide (LPS)-induced EMT. In cultured BEAS-2B cells, the protein expression levels of E-cadherin, α-smooth muscle actin (SMA), and vimentin were 0.66 ± 0.20, 1.44 ± 0.23, and 1.32 ± 0.21 in the LPS group vs 1.11 ± 0.36 (P < 0.05), 1.04 ± 0.30 (P < 0.05), and 0.96 ± 0.13 (P < 0.01) in the LPS + SP group (mean ± standard deviation), respectively. Meanwhile, LPS-triggered inflammatory cytokines and extracellular proteins were also reduced by SP administration in BEAS-2B cells. Moreover, SP treatment attenuated inflammation, EMT, extracellular matrix (ECM) deposition, and even fibrosis in a mouse EMT model. In terms of mechanism, LPS-treated BEAS-2B cells exhibited a higher level of phosphatidylinositol 3-kinase (PI3K)/AKT/mammalian target of rapamycin (mTOR) phosphorylation, which was interrupted by SP treatment. It is worth noting that the blockade of the PI3K/Akt/mTOR signaling cascade reduced the LPS-evoked EMT process in BEAS-2B cells. These results suggest that SP can block LPS-induced EMT via inhibition of the PI3K/Akt/mTOR signaling cascade, which provides a basis for possible clinical use of SP in airway and lung diseases.

Evaluating the Activity of Sodium Butyrate to Prevent Osteoporosis in Rats by Promoting Osteal GSK-3ß/Nrf2 Signaling and Mitochondrial Function.

Oxidative stress (OS) and mitochondrial dysfunction are key pathophysiological features of osteoporosis and obesity. Sodium butyrate (NaB), produced by fermentation by the gut microbiota of the large intestine, has been demonstrated to protect against OS by improving specific antioxidant enzymes and to regulate mitochondria redox homeostasis in vivo. Here, in an unblinded study, we identified femur mitochondria as the main target of the beneficial effects of NaB, consisting of reversion of bone loss and body-weight gain in obesity-prone rats. In particular, NaB promoted the activity of mitochondrial antioxidant enzymes and energy metabolism, preserved the bone microstructure and calcium homeostasis, and activated bone metabolism, as shown by increased Nrf2/GSK-3ß signaling and the upregulation of PGC-1α and TFAM. In vitro experiments showed that moderate NaB treatment prevented H2O2-induced oxidative damage in MC3T3-E1 cells, improved osteoblast mineralization and differentiation, and maintained the balance in bone metabolism by enhancing intracellular antioxidant enzyme activity and ATP production and decreasing the ROS level. In conclusion, NaB promoted the Nrf2/GSK-3ß signaling pathway and mitochondrial function and is a potential new therapeutic strategy for obesity and osteoporosis.

Deep Convolutional Radiomic Features on Diffusion Tensor Images for Classification of Glioma Grades.

The grading of glioma has clinical significance in determining a treatment strategy and evaluating prognosis to investigate a novel set of radiomic features extracted from the fractional anisotropy (FA) and mean diffusivity (MD) maps of brain diffusion tensor imaging (DTI) sequences for computer-aided grading of gliomas. This retrospective study included 108 patients who had pathologically confirmed brain gliomas and DTI scanned during 2012-2018. This cohort included 43 low-grade gliomas (LGGs; all grade II) and 65 high-grade gliomas (HGGs; grade III or IV). We extracted a set of radiomic features, including traditional texture, morphological, and novel deep features derived from pre-trained convolutional neural network models, in the manually-delineated tumor regions. We employed support vector machine and these radiomic features for two classification tasks: LGGs vs HGGs, and grade III vs IV. The area under the receiver operating characteristic (ROC) curve (AUC), accuracy, sensitivity, and specificity was reported as the performance metrics using the leave-one-out cross-validation method. When combining FA+MD, AUC = 0.93, accuracy = 0.94, sensitivity = 0.98, and specificity = 0.86 in classifying LGGs from HGGs, while AUC = 0.99, accuracy = 0.98, sensitivity = 0.98, and specificity = 1.00 in classifying grade III from IV. The AUC and accuracy remain close when features were extracted from only the solid tumor or additionally including necrosis, cyst, and peritumoral edema. Still, the effects in terms of sensitivity and specificity are mixed. Deep radiomic features derived from pre-trained convolutional neural networks showed higher prediction ability than the traditional texture and shape features in both classification experiments. Radiomic features extracted on the FA and MD maps of brain DTI images are useful for noninvasively classification/grading of LGGs vs HGGs, and grade III vs IV.

Fructo-oligosaccharides lower serum lipid levels and suppress high-fat/high-sugar diet-induced inflammation by elevating serum and gut levels of short-chain fatty acids.

OBJECTIVE: This study aimed to investigate the effects of fructo-oligosaccharides (FOS) on serum lipid levels and to determine the mechanisms underlying these effects and the potential role of inflammation. METHODS: Male C57BL/6 mice received a normal diet, a high-fat/high-sugar (HFS) diet, or an HFS diet supplemented with 10% FOS for 10 weeks. In vivo intestinal and serum short-chain fatty acid (SCFA) levels were measured by gas chromatography. In vivo serum levels of alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), 8-hydroxy-2'-deoxyguanosine (8-OHdG), and malonaldehyde (MDA) were also measured. Lipid accumulation was visualized. Reactive oxygen species (ROS) generation was evaluated and apoptosis was quantified. RESULTS: FOS reversed in vivo HFS-induced lipid accumulation in the liver. An HFS diet increased ALT, AST, TC, TG, and LDL serum levels, decreased HDL serum levels, and increased IL-6, TNF-α, 8-OHdG, and MDA levels. These changes were reduced by FOS. FOS also increased intestinal and serum levels of short chain fatty acids (SCFAs). In vitro, SCFAs ameliorated palmitic acid-induced ROS production and apoptosis of HepG2 cells. CONCLUSION: FOS supplementation lowers serum lipid levels and ameliorates HFS-induced inflammation by upregulating SCFAs.

Peptidomics Analysis Discloses That Novel Bioactive Peptides Participate in Necrotizing Enterocolitis in a Rat Model.

Necrotizing enterocolitis (NEC) is a common devastating gastrointestinal disease in premature infants, the molecular mechanisms of which have not been fully elucidated. Recently, endogenous peptides have garnered much attention owing to their role in diagnosis and treatment. However, changes in the peptide expression of NEC intestinal tissues remain poorly understood. In the present study, a comparative peptidomics profiling analysis was performed between NEC and control intestinal tissues via liquid chromatography-tandem mass spectrometry (LC-MS). In total, 103 upregulated and 73 downregulated peptides were identified in the intestinal tissues (fold change ≥ 1.5, p < 0.05). Bioinformatics analysis revealed that these differentially expressed peptides were significantly associated with NEC pathophysiology, including apoptosis, the TGF-ß signaling pathway, the Wnt signaling pathway, and the MAPK signaling pathway. Furthermore, two putative peptides could inhibit apoptosis and promote the migration of intestinal epithelial cells induced by lipopolysaccharide; these peptides were derived from the protein domains MT1 and EZRI, respectively. In conclusion, our study revealed that endogenous peptides are involved in the pathophysiologic mechanism of NEC; nevertheless, further exploration is required in this regard.

Combination of 16S rRNA and procalcitonin in diagnosis of neonatal clinically suspected sepsis.

OBJECTIVE: To investigate the application of 16S rRNA in diagnosing patients with neonatal sepsis. METHODS: We studied 60 consecutive neonatal patients with clinically suspected sepsis and 20 non-infective cases as controls. All patients were diagnosed with sepsis by clinical and experimental criteria. Clinical characteristics were recorded and 16S rRNA sequencing was conducted for all patients. The sensitivity, specificity, and accuracy of the detection methods were analyzed. RESULTS: The detection limit of 16S rRNA sequencing was 1 × 102 CFU/mL. For suspected sepsis, the positive rate of 16S rRNA detection was 93.3%, which was similar to that of procalcitonin detection (85%), and was significantly higher than that of bacterial culture (51.7%). The specificity of procalcitonin detection (74.1%) was significantly lower than that of 16S rRNA detection (100%). Moreover, the combination of 16S rRNA and procalcitonin detection showed a sensitivity of 100%, specificity of 74.1%, and accuracy of 92.0%. For proven sepsis, the sensitivity and specificity of 16S rRNA detection were both 100.0%, and those for procalcitonin were 87.1% and 87.0%, respectively. CONCLUSION: Detection of 16S rRNA has high sensitivity and specificity in diagnosing sepsis. The combination of 16S rRNA and procalcitonin has even better sensitivity with acceptable specificity.

Modulation of fat metabolism and gut microbiota by resveratrol on high-fat diet-induced obese mice.

PURPOSE: The antioxidant resveratrol (RSV) has low bioavailability and can reach the colon to access the gut microbial ecosystem. RSV administration together with high-fat diet prevented abnormal changes of intestinal microbiota. However, whether or not RSV can reshape the intestinal microbiota of obese mice and alleviate obesity-related diseases remains to be studied. This study aimed to explore the role of RSV in alleviating high-fat-induced obesity and its relationship with oxidative stress and gut microbiota. METHODS: Male C57BL/6 mice were divided into five groups and administered for 16 weeks with: standard diet (CON), high-fat diet (60% energy for lard, HFD), and HFD with low, medium, and high dose of RSV, 50, 75, and 100 mg/kg body weight administered daily via drinking water, respectively. RESULTS: Medium and high RSV treatment significantly prevented body weight gain, decreased relative weight of liver and adipose tissue compared with HFD (P<0.05). All doses significantly prevented HFD-induced increase of serum triglyceride, low density lipoprotein cholesterol, glucose, and endotoxemia (P<0.05). Medium and high dose also prevented chronic inflammation by decreasing serum interleukin-1 and tumor necrosis factor-alpha (P<0.05), and oxidative stress in liver and brain indicated by increase in superoxide dismutase, catalase, glutathione peroxidase activity (P<0.05). Formation of malondialdehyde was prevented by all doses compared with HFD (P<0.05). Both medium and high doses of RES increased alpha diversity of gut microbiota according to the Chao1 and Shannon indices (P<0.05). Medium dose induced obvious shift in gut microbiota composition according to principal component analysis. High dose of RSV effectively prevented HFD-induced increase of Coriobacteriaceae and Desulfovi-brionaceae (P<0.05), which show a significant correlation with body weight (r>0.8 P<0.00). CONCLUSION: RSV prevented HFD-induced endotoxemia, oxidative stress, and gut microbiota change.

Inhibition of HMGB1 improves necrotizing enterocolitis by inhibiting NLRP3 via TLR4 and NF-κB signaling pathways.

OBJECTIVE: To explore the relationship between high-mobility group box 1 (HMGB1) and NLR pyrin domain containing 3 (NLRP3) in the development of necrotizing enterocolitis (NEC). METHODS: NEC rat models were constructed and treated with HMGB1 inhibitor glycyrrhizin (GL) with different concentration. An inflammatory condition of intestinal tissue in newborn NEC rats was observed by hematoxylin and eosin staining. The messenger RNA (mRNA) and protein expression of HMGB1, NLRP3, toll-like receptor 4 (TLR4), nuclear factor-κB (NF-κB), and caspase 1 were determined by real-time polymerase chain reaction and western blot analysis, respectively. The content of interleukin (IL)-1ß and tumor necrosis factor-α (TNF-α) was determined by enzyme-linked immunosorbent assay. Human intestinal epithelial cell lines were induced to NEC by lipopolysaccharides (LPSs). LPS-induced cells were transfected with small interfering RNA-HMGB1 and NLRP3 plasmid vector. The mRNA and protein expression of HMGB1, NLRP3, TLR4, NF-κB, caspase 1, IL-1ß, and TNF-α were determined by real-time PCR and western blot analysis, respectively. RESULTS: The mRNA and protein expression of HMGB1 and NLRP3 in the NEC group was significantly higher than the control group. Inhibition of HMGB1 expression improved intestinal inflammation in newborn NEC rats. The expression of HMGB1, NLRP3, TLR4, NF-κB, and caspase 1 was upregulated in NEC and was weakened after treating with GL. LPS induction to intestinal epithelial cells markedly increased the expression of HMGB1, NLRP3, TLR4, NF-κB, caspase 1, IL-1ß, and TNF-α. The knockdown of HMGB1 abolished the increase of expression, whereas further transfection with NLRP3 plasmid vector recovered the increase. CONCLUSION: HMGB1 and NLRP3 were all upregulated in the development of NEC. Inhibition on HMGB1 could improve the intestinal inflammation in NEC by inhibiting NLRP3 via TLR4 and NF-κB signaling pathways.

Protective effects of γ-aminobutyric acid against H2O2-induced oxidative stress in RIN-m5F pancreatic cells.

BACKGROUND: γ-Aminobutyric acid (GABA) is a major inhibitory neurotransmitter in the central nervous system and reported to maintain the redox homeostasis and insulin secretion function of pancreatic ß cells. This study tested the hypothesis that GABA maintains cellular redox status, and modulates glycogen synthase kinase (GSK)-3ß and antioxidant-related nuclear factor erythroid 2-related factor 2 (NRF2) nuclear mass ratio in the H2O2-injured RINm5F cells. METHODS: RINm5F cells were treated with/without GABA (50, 100 and 200 µmol/L) for 48 h and then exposed to 100 µmol/L H2O2 for 30 min. Viable cells were harvested, and dichloro-dihydro-fluorescein diacetate (DCFH-DA) was used to detect reactive oxygen species (ROS) level; cellular redox status and insulin secretion were measured; cell viability was determined by 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay; mitochondrial membrane potential (MMP) was detected by flow cytometry; relative genes levels were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR); western blotting was used to determine protein expression of GSK-3ß and p-GSK-3ß (Ser9), and nuclear and cytoplasmic NRF2. RESULTS: H2O2 increased ROS production, and induced adverse affects in relation to antioxidant defense systems and insulin secretion. These changes were restored by treatment with 100 and 200 µmol/L GABA. In addition, 100 or 200 µmol/L GABA induced membrane depolarization and increased cell viability. These effects were mediated by Caspase-3, Bcl-2 associated X protein (Bax) and B-cell lymphoma-2 (Bcl-2) expression. Western blotting indicated that GABA inhibited GSK-3ß by increasing p-GSK-3ß (Ser9) level, and directed the transcription factor NRF2 to the nucleus. CONCLUSION: In rat insulin-producing RINm5F cells, GABA exerts its protective effect by regulating GSK-3ß and NRF2, which governs redox homeostasis by inhibiting apoptosis and abnormal insulin secretion by exposure to H2O2.