Gut Microbiome-Serum Metabolism Revealed the Allergenicity of Ferulic Acid Combined with Glucose-Modified ß-Lactoglobulin.

A novel strategy combining ferulic acid and glucose was proposed to reduce ß-lactoglobulin (BLG) allergenicity and investigate whether the reduction in allergenicity was associated with gut microbiome and serum metabolism. As a result, the multistructure of BLG changed, and the modified BLG decreased significantly the contents of IgE, IgG, IgG1, and mMCP-1 in serum, improved the diversity and structural composition of gut microbiota, and increased the content of short-chain fatty acids (SCFAs) in allergic mice. Meanwhile, allergic mice induced by BLG affected arachidonic acid, tryptophan, and other metabolic pathways in serum, the modified BLG inhibited the production of metabolites in arachidonic acid metabolism pathway and significantly increased tryptophan metabolites, and this contribution helps in reducing BLG allergenicity. Overall, reduced allergenicity of BLG after ferulic acid was combined with glucose modification by regulating gut microbiota, the metabolic pathways of arachidonic acid and tryptophan. The results may offer new thoughts alleviating the allergy risk of allergenic proteins.

Comparison of the Efficacy and Prognostic Factors of Endoscopic Submucosal Dissection with Different Procedures for Colorectal Neuroendocrine Tumors.

Objective: To compare the clinical efficacy, prognostic factors, and survival impact of endoscopic submucosal dissection (ESD) versus endoscopic submucosal resection (ESR) in patients with colorectal neuroendocrine tumors (NETs). Methods: This retrospective study analyzed 118 patients with colorectal NETs treated from January 2012 to December 2020. Patients were divided into the ESD group (n=59) and the ESR group (n=59) based on the surgical treatment method. We assessed the surgical efficacy, long-term survival, and factors influencing tumor recurrence using logistic regression analysis with clear criteria for group division. Results: En bloc resection, complete histological resection rates, and postoperative complications did not significantly differ between groups (P > .05). In the 33 patients with recurrence, those with tumor diameter < 10 mm, tumor grade G1, and negative resection margins were significantly fewer (P < .05). Logistic regression identified tumor diameter, grade, and resection margin status as significant predictors of recurrence (P < .05). There was no significant difference in distant metastasis, survival rates, and mortality between the groups (P > .05). Conclusions: ESD and ESR offer high clinical efficacy in treating colorectal NETs without significantly impacting prognosis or long-term survival. ESD, however, may be more suited for larger tumors due to its precise tissue removal capability. Future research should explore the long-term outcomes over 3 and 5 years to further validate these findings.

Ultrasound Improved the Non-Covalent Interaction of ß-Lactoglobulin with Luteolin: Regulating Human Intestinal Microbiota and Conformational Epitopes Reduced Allergy Risks.

The present study aims to investigate the effects of ultrasound on the non-covalent interaction of ß-lactoglobulin (ß-LG) and luteolin (LUT) and to investigate the relationship between allergenicity and human intestinal microbiota. After treatment, the conformational structures of ß-LG were changed, which reflected by the decrease in α-helix content, intrinsic fluorescence intensity and surface hydrophobicity, whereas the ß-sheet content increased. Molecular docking studies revealed the non-covalent interaction of ß-LG and LUT by hydrogen bond, van der Walls bond and hydrophobic bond. ß-LG-LUT complex treated by ultrasound has a lower IgG/IgE binding ability and inhibits the allergic reaction of KU812 cells, depending on the changes in the conformational epitopes of ß-LG. Meanwhile, the ß-LG-LUT complex affected the composition of human intestinal microbiota, such as the relative abundance of Bifidobacterium and Prevotella. Therefore, ultrasound improved the non-covalent interaction of ß-LG with LUT, and the reduction in allergenicity of ß-LG depends on conformational epitopes and human intestinal microbiota changes.

Simulated in vitro digestion of α-lactalbumin modified by phosphorylation: Detection of digestive products and allergenicity.

The effects of phosphorylation on the allergenicity of bovine α-lactalbumin (BLA) and digestive products were studied in vitro digestion. Two components with different molecular weight and conformation were obtained from natural and phosphorylated BLA. In vivo and in vitro assessment of allergenicity showed that phosphorylation prior to digestion significantly decreased the IgE/IgG binding capacity and allergic response in KU812 cells, and reduced the levels of IgG, IgE, IL-4 and histamine, with an increase in IFN-γ levels in mouse serum, depending on the changes in BLA structures, producing numerous small peptides. There were four phosphorylated sites (S22, T29, S47 and S70) in the high molecular weight components of phosphorylated BLA after digestion. These phosphorylated sites could mask the linear epitopes of digestive products, resulting in reduced allergic activity. Phosphorylation prior to digestion of dairy products can reduce the risk of anaphylaxis in patients with milk allergy to some extent.

Bovine ß-Lactoglobulin Covalent Modification by Flavonoids: Effect on the Allergenicity and Human Intestinal Microbiota.

The present study aims to investigate the structure of covalent conjugates of bovine ß-lactoglobulin (BLG) and flavonoids (luteolin, myricetin, and hyperoside), and their effect on the allergenicity and human intestinal microbiota. Covalent modification of amino acids in BLG by flavonoids was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and o-phthaldialdehyde assay. The secondary and conformational structures of BLG were changed by the covalent modification, which were determined by the circular dichroism, Fourier transform infrared spectroscopy, fluorescence spectroscopy, and UV spectroscopy. The enzyme-linked immunosorbent assay (ELISA) and cell experiments indicated that BLG covalent conjugates could reduce IgE/IgG binding capacities and suppress the allergy reactivity of RBL-2H3 cells, suggesting that the covalent modification modulated the balance of T cells. Meanwhile, covalent modification of BLG with these flavonoids can alter the diversity of human intestinal microbiota and the community abundance at phylum, family, and genus levels. The results revealed that covalent modification of BLG with flavonoids alters human intestinal microbiota, might result in the reduction of allergenicity, which could provide information for confirming the relationship between food allergy and the intestinal microbial ecosystem.

Improved antitumor activity and IgE/IgG-binding ability of α-Lactalbumin/ß-lactoglobulin induced by ultrasonication prior to binding with oleic acid.

Bovine α-lactalbumin (α-La)/ß-lactoglobulin (ß-Lg) was pretreated through ultrasonic treatment and subsequently binding with oleic acid (OA) by heat treatment. And, the antitumor activity, IgE/IgG-binding ability, and structural modifications were investigated. After α-La/ß-Lg were treated by ultrasonic prior to binding with OA, the treated α-La/ß-Lg showed high antitumor activity and IgE/IgG-binding ability, and significantly affected the structural modifications, which reflected by the reduction in α-helix content, the increase of molecular weight, intrinsic fluorescence intensity, and surface hydrophobicity. Molecular docking studies indicated that OA bound to α-La/ß-Lg by hydrogen bonds and hydrophobic interaction. Therefore, ultrasonic prior to binding with OA could improve antitumor activity and IgE/IgG-binding ability of α-La/ß-Lg as a result of structural modifications. And, ultrasonic prior to binding with fatty acid processing of milk products alone may increase the antitumor activity, this change may enhance the risk of an allergenic reaction in milk allergy patients to some extent. PRACTICAL APPLICATIONS: Fatty acids, natural ligands associated with the bovine milk proteins, and milk protein-fatty acid complex has a variety of functional applications in the food industry. This study revealed that antitumor activity, IgE/IgG-binding ability, and structural modifications of α-La/ß-Lg induced by ultrasonic prior to binding with oleic acid. It will be beneficial to understand the mechanism of the functional changes of protein. Ultrasonic prior to binding with oleic acid will be more likely to develop a practical technology to improve the functional characteristics of milk protein and design the optimal nutritional performance of milk food.

LncRNA WWOX-AS1 sponges miR-20b-5p in hepatocellular carcinoma and represses its progression by upregulating WWOX.

Increasing evidence has revealed that long noncoding RNAs (lncRNAs) emerge as pivotal regulators in diverse cancers, including hepatocellular carcinoma (HCC). This study was conducted to investigate the role of lncRNA WWOX antisense RNA 1 (WWOX-AS1) in HCC progression. Our present study illustrated that WWOX-AS1 was lowly expressed in HCC tissues and cell lines. High WWOX-AS1 expression was further confirmed to predict a favorable prognosis in HCC patients. Through functional assays, we observed that upregulated WWOX-AS1 was correlated with decreased cell proliferation, migration, epithelial to mesenchymal transition (EMT) process and increased cell apoptosis, suggesting that WWOX-AS1 exerted anti-carcinogenic role in the development of HCC. Moreover, WWOX, the nearby gene of WWOX-AS1, was found at a low level in HCC tissues and cell lines. Furthermore, there was a positive relationship between WWOX-AS1 and WWOX. Additionally, WWOX overexpression hampered cell proliferation, migration, EMT process and induced cell apoptosis in HCC. Mechanically, WWOX-AS1 was identified as a cytoplasmic RNA in HCC cells and sponged miR-20b-5p to regulate WWOX expression. Rescue assays further indicated that WWOX knockdown counteracted WWOX-AS1 overexpression-mediated suppressive function on HCC progression. Collectively, WWOX-AS1/miR-20b-5p/WWOX axis suppresses HCC tumorigenesis, hinting a potential molecular mechanism for the therapy of HCC patients.

The mechanism of the reduction in allergenic reactivity of bovine α-lactalbumin induced by glycation, phosphorylation and acetylation.

Bovine α-lactalbumin (α-Lac) allergy is a common health problem. This study assesses the allergenic reactivity and the structural properties of α-Lac after protein modification (glycation, phosphorylation and acetylation) by ELISA, cells experiment and high-resolution mass spectrometry. Three modified methods significantly reduced the IgE/IgG-binding capacity, and the release of histamine and interleukin-6, and changed the conformational structure of α-Lac. α-Lac was glycated at K13, K16, K94, K98, and K108, phosphorylated at Y18, S22, Y103, and S112, and acetylated at K13, T33, S34, T38, S47, K62, S69, S70, K108, and K114, respectively, leading to masking the linear epitopes of α-Lac. Therefore, the decrease of allergenic reactivity of α-Lac induced by glycation, phosphorylation and acetylation depends upon not only the shielding effect of their modified sites, but also the change of conformational structure. This study confirmed that protein modification was a promising method for decreasing the allergenic reactivity of allergic proteins.

Decreased expression of Beclin-1 in patients with hepatocellular carcinoma.

PURPOSE: As a conserved cellular stress response, autophagy has recently been demonstrated to be involved in the pathogenesis of several human cancers. Beclin-1 is an important autophagy gene that is abnormally expressed in a variety of human cancers. In this study, we investigated the expression of Beclin-1 in Hepatocellular Carcinoma (HCC). METHODS: A total of 83 patients with primary HCC were enrolled in this study. The expression of Beclin-1, PCNA, NET-1, Bcl-2, and Bax was measured in tissue microarray, including 83 cases of HCC and 46 adjacent non-tumor liver tissues. The association of the expression of Beclin-1 with clinicopathological features as well as PCNA, NET-1, Bcl-2, and Bax were analyzed. RESULTS: The positive rate of Beclin-1 in HCC tissues was significantly lower than that in adjacent tissues (x2 = 4.013, p=0.012). Beclin-1 expression in HCC tissues was negatively correlated with the expression of PCNA, NET-1, and anti-apoptotic protein Bcl-2, but positively correlated with pro-apoptotic protein Bax expression. Meanwhile, Beclin-1 expression was negatively correlated with HCC Edmondson grading (p=0.0058). Furthermore, Beclin-1 expression was significantly lower in HCC patients with liver cirrhosis (p=0.029) or vascular invasion (p=0.011) than those in HCC patients without cirrhosis or vascular invasion. CONCLUSION: Decreased expression of Beclin-1 was observed in HCC tissues and negatively correlated with HCC Edmondson grading, suggesting that Beclin-1 might be a valuable prognostic indicator for HCC.

Elevated Expressions of Survivin and Endoglin in Patients with Hepatic Carcinoma.

OBJECTIVE: To investigate expression profiles of survivin and endoglin in patients with hepatic carcinoma. MATERIALS AND METHODS: Cancerous tissues (hepatic carcinoma group) of 48 patients with hepatic carcinoma and adjacent noncancerous hepatic tissues (control group) were used as objects of study. Histopathological staining [hematoxylin & eosin (H&E) staining] was used to study the pathological differences in hepatic tissues between hepatic carcinoma group and control group. Moreover, survivin and endoglin protein expressions in hepatic tissues in hepatic carcinoma group and control group were detected via western blotting. Finally, Statistical Product and Service Solutions 17.0 statistical software was used to analyze the differences in survivin and endoglin expressions in hepatic tissues between hepatic carcinoma group and control group. RESULTS: H&E staining showed that histopathological features in hepatic carcinoma group were significantly different from those in control group. Compared with those in control group, the cell structure in hepatic carcinoma group was damaged, karyopyknosis was obvious, and the hepatic injury was serious. Reverse transcription-polymerase chain reaction showed that survivin and endoglin mRNA expression levels in hepatic carcinoma group were significantly increased compared with those in control group. Besides, immunofluorescence method and western blotting revealed the low expressions of survivin and endoglin proteins in tissues in control group, which were obviously lower than those in hepatic tissues in hepatic carcinoma group. Results of analyses of variance showed that the expressions of survivin and endoglin in normal hepatic tissues and cancerous tissues had statistically significant differences (p < 0.01). Furthermore, expressions of survivin and endoglin were significantly associated with histological grade, tumor size, and tumor, node, metastasis (TNM) stage. CONCLUSION: Elevated expressions of survivin and endoglin are associated with histological grade, tumor size, and TNM stage in patients with hepatic carcinoma, indicating that survivin and endoglin might be involved in the pathogenesis of hepatic carcinoma and therapeutic targeting them might be a novel approach for the treatment of hepatic carcinoma.

Expression of B Cell-Specific Moloney Murine Leukemia Virus Integration Site 1 (BMI-1) and WW Domain-Containing Oxidoreductase (WWOX) in Liver Cancer Tissue and Normal Liver Tissue.

BACKGROUND The aim of this study was to compare the expression levels of mRNA of the B cell-specific Moloney murine leukemia virus integration site 1 (BMI-1) and the WW domain-containing oxidoreductase (WWOX) genes and their protein products in tissues from patients with liver cancer with normal liver tissues from patients without liver cancer. MATERIAL AND METHODS The liver cancer group (N=56) included patients with available tissue samples of histologically confirmed liver cancer. The control group (N=24) included histologically confirmed normal liver tissue samples. Immunofluorescence staining and Western blot were used to detect and compare protein expression of Bmi-1 and WWOX in liver tissues in the liver cancer group and the control group. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect and compare mRNA expression of BMI-1 and WWOX in liver tissues in the liver cancer group and the control group. Expression levels of the protein and mRNA levels and the clinicopathological features including patient prognosis in liver cancer were evaluated statistically using analysis of variance (ANOVA). RESULTS There were significant differences in the expression levels of protein and mRNA of BMI-1 and WWOX between the liver cancer group and the control group. BMI-1 mRNA and protein expression were significantly increased, and WWOX mRNA and protein expression were significantly reduced in liver cancer tissue, compared with normal liver tissue (p<0.05). CONCLUSIONS In liver cancer tissue compared with normal liver, the expression of BMI-1 and WWOX mRNA and their protein products were upregulated and down-regulated, respectively.

Simo Decoction Stimulates Contractions of Antral Longitudinal Smooth Muscle via Multitudinous Mechanisms.

The aim of the study was to investigate Simo decoction-induced contractions of antral smooth muscles of rats and its mechanisms. The contractile responses of longitudinal strips to consecutive concentrations of Simo decoction were characterized by atropine, gallamine, 4-diphenylacetoxy-N-methylpiperidine methiodide, and adrenaline, hexamethonium, L-arginine, and nifedipine and compared with Krebs solution (control) and acetylcholine-induced contractions. Simo decoction dose-dependently increased contractions of antral strips (P = .000 vs control); its maximal effect was higher than acetylcholine (10-3 mol L-1; P < .05); Simo decoction-induced contractions were completely inhibited by atropine, 4-diphenylacetoxy-N-methylpiperidine methiodide, or 4-diphenylacetoxy-N-methylpiperidine methiodide + gallamine (P = .000 for all) but were partly suppressed by gallamine, adrenaline, hexamethonium, L-arginine, and nifedipine (P = .000 for all). Simo decoction promotes the contractions of antral strips mainly through activation of muscarinic M3 receptor, while partly through activation of M2 receptor, Ca2+ channel, nicotinic receptor, and inhibition of adrenergic receptor as well as release of nitric oxide.

Simo decoction promotes contraction of antral circular smooth muscle mainly via muscarinic M3 receptor.

ETHNOPHARMACOLOGICAL RELEVANCE: Simo Decoction (SMD), a traditional Chinese medicine, included four elements, such as Fructus aurantii, Radix aucklandiae, Semen arecae and Radix linderae. It has been used to improve gastrointestinal dysmotility in clinical practice for a long history in China. However, the explicit mechanisms are unclear. The aim of this study was to investigate the effect of SMD on contractions of antral circular smooth muscle strips of Sprague-Dawley (SD) rats and its underlying mechanism. MATERIALS AND METHODS: The antral circular strips were prepared in the organ bath under baseline or to be incubated with muscarinic receptor antagonist atropine (10(-6)M), muscarinic M3 receptor antagonist 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) (0.4×10(-6)M), muscarinic M2 receptor antagonist gallamine (10(-6)M), adrenergic receptor agonist adrenaline (10(-7)M), exogenous nitric oxide (NO) donor l-arginine (10(-4)M), nicotinic receptor antagonist hexamethonium chloride (10(-4)M) and Ca(2+) channel antagonist nifedipine (30nM), and consecutive concentrations of SMD were added to the bath to observe the strip responses. As a control, the responses of strips after administration with the same volume of Krebs solution as SMD were also noted. The strip responses to acetylcholine (10(-7)-10(-3)M) were also noted in organ bath to compare with SMD-induced contraction. RESULTS: SMD dose-dependently evoked hypercontractility of antral circular strips, and the maximal contractile effect of circular smooth muscle induced by SMD was significantly higher than that induced by acetylcholine (10(-3)M). The responses of antral circular strips to SMD were completely antagonized by atropine, 4-DAMP or 4-DAMP+gallamine, but partly inhibited by gallamine and partly suppressed by adrenaline, l-arginine, hexamethonium chloride and nifedipine. CONCLUSIONS: SMD promotes contractions of antral circular strips in rats mainly via activation of muscarinic M3 receptor, but partly via activation of muscarinic M2 receptor, Ca(2+) channel and nicotinic receptor, inhibition of adrenergic receptor and releasing of NO.

[Research progress in magnetic thermosensitive polymeric microspheres].

As a new type of functional material, magnetic thermosensitive polymeric microspheres offer high potential application in various fields, particularly in bioengineering and biomedical fields. In this review, the development of synthesis and application of magnetic thermosensitive polymeric microspheres was summarized, and the research trends were also discussed.

[Environmental stimuli-sensitive biodegradable drug delivery systems].

Environmental stimuli-sensitive biodegradable drug delivery systems are drawing more and more attentions because of their advantages such as smart properties, high efficiency and easy-to-handle properties. On the basis of a large quantity of references on this topic, a review has been made on the developments of the thermosensitive and pH-sensitive intelligent polymeric systems for drug delivery.

[Inhibitory effect of kaempferol against binding of platelet activating factor to its receptor].

OBJECTIVE: To observe the platelet activating factor (PAF) antagonistic effect of kaempferol. METHOD: The specific binding of [3H] PAF to rabbit platelet receptor was investigatedwith radio ligand binding assay (RLBA). Platelet adhesion induced by PAF was measured with spectrophotometry. The elevation of inner free calcium concentration in rabbit polymorphonuclear leukocytes (PMNs) induced by PAF was determined with Fura-2 fluorescent technique. RESULT: The 1, 2 or 4 nmol x L(-1) [3H]PAF specific binding to rabbit platelet receptor was inhibited by Kae dosage dependently and the IC50 were 30.8, 74.6 and 92.0 micro mol x L(-1), respectively. The PAF induced reactions of rabbit platelet adhesion and PMNs inner free calcium concentration elevation were inhibited by Kae in a dose-dependent manner. The IC50 of Kae to inhibit platelet adhesion was 65 micromol x L(-1). CONCLUSION: Kae is effective in inhibiting the action of PAF and it is a new PAF receptor antagonist.

Preparation of glucose-sensitive microcapsules with a porous membrane and functional gates.

A glucose-sensitive microcapsule with a porous membrane and with linear-grafted polyacrylic acid (PAAC) chains and covalently bound glucose oxidase (GOD) enzymes in the membrane pores acting as functional gates was successfully prepared. Polyamide microcapsules with a porous membrane were prepared by interfacial polymerization, PAAC chains were grafted into the pores of the microcapsule membrane by plasma-graft pore-filling polymerization, and GOD enzymes were immobilized onto the PAAC-grafted microcapsules by a carbodiimide method. The release rates of model drug solutes from the fabricated microcapsules were significantly sensitive to the existence of glucose in the environmental solution. In solution, the release rate of either sodium chloride or VB(12) molecules from the microcapsules was low but increased dramatically in the presence of 0.2mol/L glucose. The prepared PAAC-grafted and GOD-immobilized microcapsules showed a reversible glucose-sensitive release characteristic. The proposed microcapsules provide a new mode for injection-type self-regulated drug delivery systems having the capability of adapting the release rate of drugs such as insulin in response to changes in glucose concentration, which is highly attractive for diabetes therapy.

Preparation of submicrometer-sized monodispersed thermoresponsive core-shell hydrogel microspheres.

We have successfully prepared monodispersed thermoresponsive core-shell hydrogel microspheres with a mean diameter of 200-400 nm with poly(N-isopropylacrylamide-co-styrene) [P(NIPAM-co-St)] cores and poly(N-isopropylacrylamide) (PNIPAM) shells. The submicrometer-sized monodispersed P(NIPAM-co-St) core seeds were prepared by using a surfactant-free emulsion polymerization method, and the PNIPAM shell layers were fabricated onto the core seeds by using a seed polymerization method. The particle size, morphology and monodispersity, and thermoresponsive characteristics of the prepared microspheres were experimentally studied. In the preparation of P(NIPAM-co-St) seeds, with increasing the initiator dosage, the mean diameters and the dispersal coefficients were almost at the same levels at first; however, when the initiator dosage increased further to a critical amount, the mean diameters decreased drastically and the monodispersity became worse significantly. With increasing the stirring rate, the particle diameter decreased, and when the stirring rate was larger than 600 rpm, the monodispersity became worse obviously. With increasing the phase ratio, the mean diameter became larger simply, and the monodispersity became worse first and then became better again. With increasing the reaction time, the particle sizes nearly did not change, while the monodispersity gradually became better slightly. For the core-shell microspheres, with increasing the NIPAM dosage in the preparation of the PNIPAM shell layers, the mean diameters became larger simply, the monodispersity became better, and the thermoresponsive swelling ratio of the hydrodynamic diameters increased.

[Intelligent polymeric systems for glucose-responsive insulin delivery].

Polymeric drug delivery system for insulin controlled-release is one of the most active fields of research and development in the world. Up to date, several kinds of intelligent drug carriers for glucose-responsive insulin delivery have been reported. On the basis of a large quantity of references on this topic, a review has been made on the developments of the intelligent polymeric systems for glucose-responsive insulin delivery.