Effects of an Amino Acid-Based Formula Supplemented with Two Human Milk Oligosaccharides on Growth,Tolerability, Safety,and Gut Microbiome in Infants with Cow’s Milk Protein Allergy

This open-label, non-randomized, multicenter trial (Registration: {"type":"clinical-trial","attrs":{"text":"NCT 03661736","term_id":"NCT03661736"}}NCT 03661736) aimed to assess if an amino acid-based formula (AAF) supplemented with two human milk oligosaccharides (HMO) supports normal growth and is well tolerated in infants with a cow’s milk protein allergy (CMPA). Term infants aged 1–8 months with moderate-to-severe CMPA were enrolled. The study formula was an AAF supplemented with 2′-fucosyllactose (2′-FL) and lacto-N-neotetraose (LNnT). Infants were fed the study formula for 4 months and were offered to remain on the formula until 12 months of age. Tolerance and safety were assessed throughout the trial. Out of 32 infants (mean age 18.6 weeks; 20 (62.5%) male), 29 completed the trial. During the 4-month principal study period, the mean weight-for-age Z score (WAZ) increased from –0.31 at the baseline to +0.28 at the 4-months’ follow-up. Linear and head growth also progressed along the WHO child growth reference, with a similar small upward trend. The formula was well tolerated and had an excellent safety profile. When comparing the microbiome at the baseline to the subsequent visits, there was a significant on-treatment enrichment in HMO-utilizing bifidobacteria, which was associated with a significant increase in fecal short-chain fatty acids. In addition, we observed a significant reduction in the abundance of fecal Proteobacteria, suggesting that the HMO-supplemented study formula partially corrected the gut microbial dysbiosis in infants with CMPA.     

An Efficient Dopant for Introducing Magnetism into Topological Insulator Bi2Se3

In this work, we obtained an effective way to introduce magnetism into topological insulators, and successfully fabricated single crystal C-Bi₂Se₃. The structural, electrical and magnetic properties of non-magnetic element X (B, C and N) doped at Bi, Se1, Se2 and VDW gap sites of Bi₂Se₃ were studied by the first principles. It is shown that the impurity bands formed inside the bulk inverted energy gap near the Fermi level with C doping Bi₂Se₃. Due to spin-polarized ferromagnetic coupling, the time inversion symmetry of Bi₂Se₃ is destroyed. Remarkably, C is the most effective dopant because of the magnetic moment produced by doping at all positions. The experiment confirmed that the remnant ferromagnetism Mr is related to the C concentration. Theoretical calculations and experiments confirmed that carbon-doped Bi₂Se₃ is ferromagnetic, which provides a plan for manipulating topological properties and exploring spintronic applications.     

A Novel Hypoxia-inducible Factor 1α Inhibitor KC7F2 Attenuates Oxygen-induced Retinal Neovascularization

PurposeKC7F2 is a novel molecule compound that can inhibit the translation of hypoxia-inducible factor 1α (HIF1α). It has been reported to exhibit potential antiangiogenic effect. We hypothesized that KC7F2 could inhibit oxygen-induced retinal neovascularization (RNV). The purpose of this study was to investigate this assumption.MethodsOxygen-induced retinopathy (OIR) models in C57BL/6J mice and Sprague-Dawley rats were used for in vivo study. After intraperitoneal injections of KC7F2, RNV was detected by immunofluorescence and hematoxylin and eosin staining. Retinal inflammation was explored by immunofluorescence. EdU incorporation assay, cell counting kit-8 assay, scratch test, transwell assay, and Matrigel assay were used to evaluate the effect of KC7F2 on the proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVEC) induced by vascular endothelial growth factor (VEGF) in vitro. Protein expression was examined by Western blot.ResultsKC7F2 treatment (10 mg/kg/d) in OIR mice significantly attenuated pathological neovascularization and decreased the number of preretinal neovascular cell nuclei, without changing the avascular area, which showed the same trends in OIR rats. Consistently, after the KC7F2 intervention (10 µM), cell proliferation was inhibited in VEGF-induced HUVEC, which was in agreement with the trend observed in the retinas of OIR mice. Meanwhile, KC7F2 suppressed VEGF-induced HUVEC migration and tube formation, and decreased the density of leukocytes and microglia colocalizing neovascular areas in the retinas. Moreover, the HIF1α–VEGF pathway activated in retinas of OIR mice and hypoxia-induced HUVEC, was suppressed by KC7F2 treatment.ConclusionsThe current study revealed that KC7F2 was able to inhibit RNV effectively via HIF1α–VEGF pathway, suggesting that it might be an effective drug for RNV treatment.     

Suppression of Anti-Inflammatory Mediators in Metabolic Disease May Be Driven by Overwhelming Pro-Inflammatory Drivers

Obesity is a multifactorial disease and is associated with an increased risk of developing metabolic syndrome and co-morbidities. Dysregulated expansion of the adipose tissue during obesity induces local tissue hypoxia, altered secretory profile of adipokines, cytokines and chemokines, altered profile of local tissue inflammatory cells leading to the development of low-grade chronic inflammation. Low grade chronic inflammation is considered to be the underlying mechanism that increases the risk of developing obesity associated comorbidities. The glucocorticoid induced protein annexin A1 and its N-terminal peptides are anti-inflammatory mediators involved in resolving inflammation. The aim of the current study was to investigate the role of annexin A1 in obesity and associated inflammation. To achieve this aim, the current study analysed data from two feasibility studies in clinical populations: (1) bariatric surgery patients (Pre- and 3 months post-surgery) and (2) Lipodystrophy patients. Plasma annexin A1 levels were increased at 3-months post-surgery compared to pre-surgery (1.2 ± 0.1 ng/mL, n = 19 vs. 1.6 ± 0.1 ng/mL, n = 9, p = 0.009) and positively correlated with adiponectin (p = 0.009, r = 0.468, n = 25). Plasma annexin A1 levels were decreased in patients with lipodystrophy compared to BMI matched controls (0.2 ± 0.1 ng/mL, n = 9 vs. 0.97 ± 0.1 ng/mL, n = 30, p = 0.008), whereas CRP levels were significantly elevated (3.3 ± 1.0 µg/mL, n = 9 vs. 1.4 ± 0.3 µg/mL, n = 31, p = 0.0074). The roles of annexin A1 were explored using an in vitro cell based model (SGBS cells) mimicking the inflammatory status that is observed in obesity. Acute treatment with the annexin A1 N-terminal peptide, AC2-26 differentially regulated gene expression (including PPARA (2.8 ± 0.7-fold, p = 0.0303, n = 3), ADIPOQ (2.0 ± 0.3-fold, p = 0.0073, n = 3), LEP (0.6 ± 0.2-fold, p = 0.0400, n = 3), NAMPT (0.4 ± 0.1-fold, p = 0.0039, n = 3) and RETN (0.1 ± 0.03-fold, p < 0.0001, n = 3) in mature obesogenic adipocytes indicating that annexin A1 may play a protective role in obesity and inflammation. However, this effect may be overshadowed by the continued increase in systemic inflammation associated with rapid tissue expansion in obesity.     

The Role of Phytosterols in Nonalcoholic Fatty Liver Disease

Nonalcoholic fatty liver disease is now recognized as the most common cause of chronic liver disease with an increasing prevalence in both adults and children. Although the symptoms are absent or poorly expressed in most cases, some patients may progress to end-stage liver disease. The pathogenesis of NAFLD is known to be multifactorial. Current therapeutic recommendations focus on lifestyle changes in order to reduce the incidence of risk factors and drugs targeting major molecular pathways potentially involved in the development of this disease. Given that a pharmacological treatment, completely safe and effective, is not currently known in recent years more research has been done on the effects that some bio-active natural compounds, derived from plants, have in preventing the onset and progression of NAFLD. Numerous studies, in animals and humans, have shown that phytosterols (PSs) play an important role in this pathology. Phytosterols are natural products that are found naturally in plant. More than 250 phytosterols have been identified, but the most common in the diet are stigmasterol, β-sitosterol, and campesterol. Consumption of dietary PSs can reduce serum cholesterol levels. Due to these properties, most studies have focused on their action on lipid metabolism and the evolution of NAFLD. PSs may reduce steatosis, cytotoxicity oxidative stress, inflammation, and apoptosis. The purpose of this review is to provide an overview of the importance of dietary phytosterols, which are a window of opportunity in the therapeutic management of NAFLD.     

褐藻素通过调控Nrf2/Keap1通路缓解糖尿病大鼠心肌肥大

目的探索褐藻素（FX）对糖尿病心肌病的保护效应和作用机制。方法腹腔注射链脲佐菌素建立糖尿病大鼠模型，进行分组：糖尿病组（DM）、褐藻素干预糖尿病组（DM+FX）和二甲双胍干预糖尿病组（DM+Met），另取正常大鼠给予正常喂养作为正常组（Con）。造模成功后连续12周每日灌胃给药，褐藻素组每日给予200 mg/kg褐藻素灌胃，二甲双胍组每日给予230 mg/kg灌胃，糖尿病组同步灌胃生理盐水。HE染色观察各组大鼠心脏细胞肥大情况；Western blot法检测大鼠心脏中纤维化蛋白TGF-β1和FN蛋白表达水平。H9C2细胞分为3组：正常糖组（NG，5.5 mmol/L葡萄糖）、高糖组（HG，45 mmol/L葡萄糖）和褐藻素干预高糖组（HG+1 μmol/L FX）。FITC标记的鬼笔环肽检测大鼠心肌细胞H9C2表面积变化；qRT-PCR法测定各组细胞肥大因子ANP、BNP和β-MHC基因表达变化；Western blot法检测各组大鼠心脏组织和H9C2细胞中Nrf2、Keap1、HO-1、SOD1蛋白表达水平；DCFH-DA探针检测细胞内活性氧水平变化。结果褐藻素改善糖尿病大鼠心肌纤维化和心肌细胞肥大，同时上调心脏组织中Nrf2和HO-1蛋白表达，抑制Keap1蛋白表达（P < 0.05）。褐藻素可抑制高糖诱导H9C2心肌细胞表面积增加，降低ANP、BNP和β-MHC的mRNA表达水平（P < 0.05）。褐藻素可促进高糖诱导的H9C2细胞中Nrf2入核，上调下游靶蛋白SOD1和HO-1蛋白表达（P < 0.05）来增强细胞抗氧化能力，降低细胞内活性氧的水平。结论褐藻素具有良好的抗糖尿病心肌纤维化和心肌细胞肥大作用，同时上调Nrf2信号通路并促进下游抗氧化蛋白SOD1和HO-1的表达，降低活性氧水平。     

干扰肿瘤相关巨噬细胞的P2X4受体表达可抑制胶质瘤细胞的迁移和侵袭

目的研究P2X4受体在小鼠胶质瘤中肿瘤相关巨噬细胞（TAMs）的表达对胶质瘤细胞侵袭迁移的影响及分子机制。方法将16只健康C57BL/6小鼠随机分为肿瘤组、对照组（8只/组），利用小鼠胶质瘤GL261细胞接种于小鼠大脑尾状核，建立小鼠脑胶质瘤模型。术后第21天处死小鼠，取荷瘤小鼠及正常小鼠脑组织，采用HE染色观察小鼠胶质瘤形态，利用免疫荧光检测Iba-1、P2X4受体的表达情况；应用GL261条件培养基将RAW264.7细胞诱导为TAMs，采用RT-qPCR检测巨噬细胞极化相关标志物及P2X4受体在TAMs的mRNA表达情况；采用Western blot检测P2X4受体在TAMs的蛋白表达情况；采用siRNA P2X4，下调TAMs中P2X4受体的蛋白表达，RT-qPCR、Western blot检测siRNA P2X4转染后TAMs中IL-1β、IL-18的mRNA及蛋白表达；利用transwell侵袭及迁移实验检测siRNA P2X4转染后TAMs对GL261细胞侵袭迁移能力的影响。结果与对照组相比，荷瘤小鼠脑胶质瘤组织中有较高数量的Iba-1阳性细胞（P < 0.0001），且P2X4受体在Iba-1阳性细胞中的表达增高（P=0.001）；使用GL261条件培养基刺激RAW264.7细胞转化为TAMs后，M2型巨噬细胞标志基因Arg-1、IL-10表达上调（P=0.0001、0.001），M1型巨噬细胞标志基因iNOS、TNF-α表达也上调（P=0.006、0.001），但以M2型巨噬细胞标志基因Arg-1、IL-10表达上调更为显著，同时TAMs中P2X4受体蛋白表达水平及mRNA水平均增高（P=0.005、0.014）。干扰TAMs中P2X4受体表达引起其IL-1β、IL-18的mRNA（P < 0.01）及蛋白表达水平（P < 0.01，P < 0.05）降低，并抑制TAMs促进胶质瘤细胞侵袭和迁移的能力（P=0.004、0.017）。结论降低肿瘤相关巨噬细胞P2X4受体表达可能通过IL-1β、IL-18影响胶质瘤细胞的迁移和侵袭。     

β-catenin inhibitors ICG-001 and pyrvinium sensitize bortezomib-resistant multiple myeloma cells to bortezomib

Although bortezomib (BTZ) displays efficacy in treating multiple myeloma (MM), BTZ resistance in MM patients has been reported. Meanwhile, treating BTZ resistant MM cells with β-catenin inhibitors have demonstrated the ability to reserve BTZ resistance. Thus, the present study aimed to investigate the synergistic effect of the β-catenin inhibitors, ICG-001 and pyrvinium (PP), with BTZ in the treatment of BTZ-resistant MM cells. Different concentrations of ICG-001 (0–32 µM) or PP (0–32 nM) were used to treat the BTZ-resistant RPMI-8226 (RPMI-8226BR) and BTZ-resistant KMS-11 (KMS-11BR) cell lines, followed by a BTZ combination treatment. Subsequently, cell viability and apoptosis in these two cell lines were determined by CCK-8 assay and flow cytometry, respectively. The proteins involved in the Wnt/β-catenin signaling pathway were detected using western blotting. The Wnt/β-catenin signaling pathway was activated in the RPMI-8226BR and the KMS-11BR cells. In addition, the cell viability of RPMI-8226BR and KMS-11BR cells were decreased following β-catenin inhibitor (ICG-001 and PP) treatment alone. Furthermore, the β-catenin inhibitors, ICG-001 and PP, plus BTZ combination treatment revealed a notable decrease in cell viability and a marked increase in cell apoptosis rate, compared with that in cells treated with ICG-001, PP or BTZ alone in the RPMI-8226BR and KMS-11BR cell lines. In conclusion, the β-catenin inhibitors, ICG-001 and PP not only increased apoptosis, but also sensitized BTZ-resistant MM cells to BTZ, indicating their potential therapeutic application in MM.