Identification and Function Analysis of Novel Hypoglycemic and Antioxidant Peptides from Chickpea.

Chickpea is rich in protein and has been demonstrated to possess hypoglycaemic effects. However, the specific bioactive ingredients and mechanisms underlying their hypoglycaemic effects remain unclear. In this study, enzymatic hydrolysis and gel permeation chromatography were used to extract chickpea bioactive peptide (CBP) from chickpea protein. One of the products, CBP-75-3, was found to inhibit α-glucosidase (GAA) activity and significantly increase the viability of insulin resistant (IR) cells. Moreover, CBP-75-3 significantly increased the rate of glucose consumption and glycogen synthesis in IR-HepG2 cells. Moreover, CBP-75-3 decreased the levels of malondialdehyde and increased the levels of superoxide dismutase, glutathione, and glutathione peroxidase. Subsequently, 29 novel bioactive peptides in CBP-75-3 were identified by LC‒MS/MS, and the potential hypoglycaemic targets of these novel bioactive peptides were investigated using molecular docking. Based on the results, the residues of the novel bioactive peptides interact with GAA through hydrogen bonding (especially LLR, FH, RQLPR, KGF and NFQ by binding to the substrate binding pocket or the active centre of GAA), thereby inhibiting GAA activity and laying a foundation for its hypoglycaemic activity. In short, the novel bioactive peptides isolated and identified from chickpea can effectively exert hypoglycaemic effects and increase the antioxidant capacity of IR-HepG2 cells. This study reveals that CBP-75-3, a natural hypoglycaemic ingredient, has potential for applications in functional foods and provides a theoretical basis for the development and application of CBP in the future.

Iso-E-Codelock: A Rebuilding-free Electrochemical Chip with a Customizable Decoding Probe for Real-Time and Portable Pathogen Diagnostics.

In order to realize portable pathogen diagnostics with easier quantitation, digitization and integration, we develop a ready-to-use electrochemical sensing strategy (Iso-E-Codelock) for real-time detection of isothermal nucleic acid amplification. Bridged by a branched DNA as codelock, the isothermal amplicon is transduced into increased current of an electrochemical probe, holding multiple advantages of high sensitivity, high selectivity, signal-on response, "zero" background and one-pot operation. Through a self-designed portable instrument (BioAlex PHE-T), the detection can be implemented on a multichannel microchip and output real-time amplification curves just like an expensive commercial PCR machine. The microchip is a rebuilding-free and disposable component. The branch codelock probe can be customized for different targets and designs. Such high performance and flexibility have been demonstrated utilizing four virus (SARS-CoV-2, African swine fever, FluA and FluB) genes as targets, and two branch (3-way and 4-way) DNAs as codelock probes.

Pogostemon cablin (Blanco) Benth granule revealed a positive effect on improving intestinal barrier function and fecal microbiota in mice with irinotecan-induced intestinal mucositis.

Pogostemon cablin (Blanco) Benth (PCB), a medicinal and edible homologous Chinese herb, has a protective effect on the structure and function of intestine. In this study, we aimed to investigate the effect of PCB granule (PCBG) on the improvement of irinotecan-induced intestinal mucositis and the regulation of intestinal microorganisms in mice. Our results demonstrated that PCBG supplementation significantly improved diarrhea symptoms caused by irinotecan, as evidenced by inhibiting weight loss, reversing intestinal atrophy, protecting against splenomegaly and balancing oxidative stress. Furthermore, compared with the model group, PCBG restored the intestinal morphology and improved intestinal barrier dysfunction by promoting the expression of tight junction proteins and mucin. Moreover, high-throughput sequencing analysis revealed that PCBG improved the flora disorder caused by irinotecan and regulated microbial community structure, such as decreasing the relative abundance of Bacteroides as well as increasing the relative abundance of Lactobacillus. Meanwhile, the disordered microbial functions in intestinal mucositis mice were recovered more closely to the controls by PCBG. Finally, we found that a robust correlation between the specific microbiota and intestinal mucositis-related index. In summary, these findings revealed the beneficial effects of PCBG on the intestinal barrier and gut microbiota of irinotecan-induced intestinal mucositis, which may be one of the potential strategies to reduce the clinical side effects of irinotecan.

Analysis of Prognostic Factors of Low-Grade Gliomas in Adults Using Time-Dependent Competing Risk Models: A Population Study Based on the Surveillance, Epidemiology, and End Results Database.

BACKGROUND: Because of multiple competing death outcomes and time-varying coefficients, using a Cox regression model to analyze the prognostic factors of low-grade gliomas (LGG) may lead to a possible bias. Therefore, we adopted time-dependent competing risk models to obtain accurate prognostic factors for LGG. METHODS: In this retrospective cohort study, data were extracted from patients enrolled in the Surveillance, Epidemiology, and End Results (SEER) database between 2000 and 2018. Univariate analysis was performed using the cumulative incidence function (CIF) and Kaplan-Meier (KM) function. Time-dependent competing risk and Cox regression models were used in the multivariable analysis. RESULTS: A total of 2581 patients were diagnosed with low-grade glioma, among whom 889 died from low-grade glioma, 114 died from other causes, and the rest were alive. The time-dependent competing risk models indicated that age, sex, marital status, primary tumor site, histological type, tumor diameter, surgery, and year of diagnosis were significantly associated with low-grade glioma-specific death, and the relative effect of age, tumor diameter, surgery, oligodendroglioma, and mixed glioma on low-grade glioma-specific death changed over time. Compared with the competing risk models, the Cox regression model misestimated the hazard ratio (HR) of covariates on the outcome and even produced false-negative results. CONCLUSIONS: The time-dependent competing risk models were better than the Cox regression model for evaluating the impact of covariates on low-grade glioma-specific mortality in the presence of competing risks and time-varying coefficients. The models identified the prognostic factors of LGG more accurately than the Cox regression model.

Treatment options for patients with CADASIL and large-scale cerebral infarction: mechanical thrombectomy and antiplatelet therapy-A case report.

Background: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an autosomal dominant inherited arterial disease, with lacunar infarction resulting from intracranial small vessel lesions being the most prevalent clinical manifestation of CADASIL. However, large-scale cerebral infarction caused by intracranial non-small vessels occlusion is relatively uncommon, and reports of vascular intervention and long-term antiplatelet drug treatment for patients with CADASIL and large-scale cerebral infarction are rarer. Methods: We reported a 52 year-old male who experienced a significant cerebral infarction due to an occlusion in the second segment of the left middle cerebral artery, 4 months subsequent to being diagnosed with CADASIL. Following the benefit and risk assessment, the patient underwent intracranial vascular thrombectomy and balloon dilation angioplasty. Subsequently, he was administered dual antiplatelet therapy for 3 months, followed by mono antiplatelet therapy. Results: After undergoing intracranial vascular intervention and receiving antiplatelet therapy, significant improvement in the symptoms were observed. The National Institutes of Health Stroke Scale score decreased from 6 to 2 points, and no bleeding lesions were detected on the head computed tomography during regular follow-up visits after discharge. Conclusion: Our case highlights the possibility that patients with CADASIL may also encounter extensive cerebral infarction resulting from stenosis or occlusion of intracranial non-small vessels. Considering the specific circumstances of the patient, intravascular intervention and antiplatelet therapy can be regarded as viable treatment options for individuals with CADASIL.

Identification and screening of potential anti-pneumonia active ingredients and targets of Qing-Kai-Ling oral liquid via UHPLC-Q-Exactive Orbitrap mass spectrometry based on data post-processing.

Qing-Kai-Ling oral liquid is commonly used clinically for the treatment of fever and upper respiratory tract infection. Moreover, studies have shown that Qing-Kai-Ling oral liquid has an anti-pneumonia effect. However, owing to its complex pharmacodynamic material basis, its pharmacological research and clinical application are limited. To address this problem, the chemical constituents of Qing-Kai-Ling oral liquid were identified by ultra-high performance liquid chromatography quadrupole-Exactive Orbitrap mass (UHPLC-Q-Exactive Orbitrap MS) and network pharmacology methods, which were used to predict its potential anti-pneumonia target and signalling pathway. A total of 150 compounds were identified and tentatively characterized, including 35 amino acids and their derivatives, 36 organic acids, 20 terpenoids, 20 alkaloids, 12 glycosides, 7 flavonoids, and 20 others. Among them, 14 compounds were accurately identified by comparing their retention time and mass spectrum data with those of reference substances. Additionally, we performed molecular simulation calculations via Density Function Theory to determine the plausibility of the compound cleavage reactions and further confirm compound structures. Furthermore, 90 key targets were screened through network pharmacology, with the particular focus on the PI3K-AKT, MAPK and TNF signalling pathways. This method achieved the first comprehensive identification of the chemical composition of Qing-Kai-Ling oral liquid and elucidated its potential mechanism of anti-pneumonia. The results provide valuable reference and data support for pharmacodynamic substance research and quality control.

Lactobacillus acidophilus and its metabolite ursodeoxycholic acid ameliorate ulcerative colitis by promoting Treg differentiation and inhibiting M1 macrophage polarization.

Lactobacillus acidophilus (LA) is a common clinical probiotic that improves ulcerative colitis (UC) by restoring intestinal immune balance. However, the interaction of LA with the gut microbiota and its metabolites in the treatment of UC remains unknown. Therefore, this study seeks to elucidate whether the gut microbiota and its metabolites act as pivotal effectors in LA's therapeutic mechanisms and how precisely they modulate intestinal immunity. In this study, we verified that LA can obviously ameliorate the disease severity, and regulate intestinal immune disorders in UC mice. Subsequently, antibiotic (ABX)-mediated depletion of the gut microflora demonstrated that the therapeutic efficiency of LA was closely associated with gut microbiota. In addition, the results of metabolomics revealed that ursodeoxycholic acid (UDCA), a metabolite of intestinal flora, may be a potential effector molecule mediating therapeutic effects of LA. Indeed, we found that UDCA can improve the macro pathological characteristics of UC mice, and through a comprehensive set of in vivo and in vitro experiments, we discovered that UDCA exerts dual effects on immune regulation. Firstly, it promotes the differentiation of Treg cells, resulting in increased secretion of anti-inflammatory cytokines. Secondly, UDCA inhibits the polarization of M1 macrophages, effectively reducing the secretion of pro-inflammatory cytokines. Moreover, we found that UDCA regulation of immune response is directly related to the RapGap/PI3K-AKT/NF-κB signaling pathway. In conclusion, LA and its metabolite, UDCA, may treat UC by activating the RapGap/PI3K-AKT/NF-κB signaling pathway and modulating Treg cells and M1 macrophages. All in all, our findings highlight the potential of microbial metabolites in enhancing probiotic for UC treatment.

Rapidly enrichment and detection of per-and polyfluoroalkyl substances in foods using a novel bifunctional covalent organic framework.

Per- and polyfluoroalkyl substances (PFASs) are extensively found in foods, posing potential toxicity to humans. Therefore, rapid analysis and monitoring of PFASs in foods are crucial for public health and also a challenge. To detect trace PFASs in foods, construction of sorbents with multiple interactions could be an effective approach. Herein, a cationic-fluorinated covalent organic framework (CF-COF) was prepared by post-modification and used as a magnetic solid-phase extraction adsorbent for adsorption of PFASs. By combining magnetic solid-phase extraction based on CF-COF with liquid chromatography-tandem mass spectrometry (LC - MS/MS), a novel method was developed for determination of eight long-chain PFASs in foods. Under optimized conditions, the method exhibited low detection limits (0.003-0.019 ng/g) and satisfactory recovery rates (73.5-118%) for PFASs. This study introduces a novel idea for the development of adsorbents targeting PFASs, along with a new analytical method for monitoring of PFASs in foods.

FUS Selectively Facilitates circRNAs Packing into Small Extracellular Vesicles within Hypoxia Neuron.

Small extracellular vesicles (sEVs) contain abundant circular RNAs (circRNAs) and are involved in cellular processes, particularly hypoxia. However, the process that packaging of circRNAs into neuronal sEVs under hypoxia is unclear. This study revealed the spatial mechanism of the Fused in Sarcoma protein (FUS) that facilitates the loading of functional circRNAs into sEVs in hypoxia neurons. It is found that FUS translocated from the nucleus to the cytoplasm and is more enriched in hypoxic neuronal sEVs than in normal sEVs. Cytoplasmic FUS formed aggregates with the sEVs marker protein CD63 in cytoplasmic stress granules (SGs) under hypoxic stress. Meanwhile, cytoplasmic FUS recruited of functional cytoplasmic circRNAs to SGs. Upon relief of hypoxic stress and degradation of SGs, cytoplasmic FUS is transported with those circRNAs from SGs to sEVs. Validation of FUS knockout dramatically reduced the recruitment of circRNAs from SGs and led to low circRNA loading in sEVs, which is also confirmed by the accumulation of circRNAs in the cytoplasm. Furthermore, it is showed that the FUS Zf_RanBP domain regulates the transport of circRNAs to sEVs by interacting with hypoxic circRNAs in SGs. Overall, these findings have revealed a FUS-mediated transport mechanism of hypoxia-related cytoplasmic circRNAs loaded into sEVs under hypoxic conditions.

16S rRNA and Metagenomics Combined with UPLC-Q/TOF-MS Metabolomics Analysis Reveals the Potential Mechanism of Radix Astragali Against Hyperuricemia in Mice.

Purpose: This study aimed to investigate the underlying treatment mechanism of Radix Astragali (RA) in hyperuricemia from the perspective of microbiota and metabolomics. Methods: We used potassium oxyazinate (PO) to induce hyperuricemia mice, and we determined serum alanine aminotransferase/aspartate aminotransferase (ALT/AST), xanthine oxidase (XOD), creatinine (CRE), uric acid (UA), blood urea nitrogen (BUN) levels, liver XOD levels and assessed the kidney tissue histopathology. The therapeutic mechanism of RA in hyperuricemic mice was studied by 16S rRNA, metagenomic sequencing and metabolomics. Results: Our research showed that RA has therapeutic effect in hyperuricemia mice, such as slow the weight loss, repair kidney damage, and downregulate serum UA, XOD, CRE, ALT/AST, BUN, and liver XOD levels. RA restored the disturbance structure of the microbiota in hyperuricemia mice by increasing the relative abundances of beneficial bacteria (Lactobacillaceae and Lactobacillus murine) but decreasing the relative abundances of pathogenic bacteria (Prevotellaceae, Rikenellaceae and Bacteroidaceae). Meanwhile, we found that RA directly regulated the metabolic pathway (such as linoleic acid metabolism and glycerophospholipid metabolism) and indirectly regulated bile acid metabolism by mediating microbiota to ameliorate metabolic disorders. Subsequently, there was a robust correlation between specific microbiota, metabolites and the disease index. Conclusion: The ability of RA to protect mice against hyperuricemia is strongly linked to the microbiome-metabolite axis, which would provide evidence for RA as a medicine to prevent or treat hyperuricemia.

Kidney tea ameliorates hyperuricemia in mice via altering gut microbiota and restoring metabolic profile.

Clerodendranthus spicatus (Thunb.) C. Y. Wu, also known as kidney tea (KT), has been widely employed in kidney protection in Chinese Medicine. It has been reported that KT can lower uric acid (UA) and mitigate gout, while the mechanism remains to be elucidated. Given the close relationship between hyperuricemia (HUA), intestinal flora and host metabolism, this study aimed to explore the mechanism by which KT lowers UA from the perspective of the fecal microbiome and metabolome. Initially, mice were intraperitoneally injected with potassium oxonate to induce the HUA model. The results showed that KT markedly reduced the serum level of UA and impaired renal damage in HUA mice. Subsequently, the result of 16S rRNA gene sequencing analysis indicated that KT administration appeared a significant improvement in the structure of the intestinal flora, especially increased the abundances of Roseburia and Enterorhabdus, while decreased the abundances of Ileibacterium and UBA1819. Moreover, the levels of differential metabolites (including twenty-five in feces and eight in serum) identified by untargeted metabolomics returned to normal after KT intervention. Taken together, the mechanism of KT in alleviating HUA is related to the regulation of the intestinal flora and the remodeling of metabolic disorders, which will lay a theoretical foundation for KT as a UA-lowering drug.

Live Lactobacillus acidophilus alleviates ulcerative colitis via the SCFAs/mitophagy/NLRP3 inflammasome axis.

As a disease caused by an impaired intestinal epithelial barrier, imbalanced flora, immune imbalance and genetic susceptibility, ulcerative colitis (UC) is becoming a health threat for all ages. Lactobacillus acidophilus (L. acidophilus), an attracting probiotic, has already been confirmed to improve immune dysfunction, stabilize intestinal microflora, and combat gut disorders. However, no studies have focused on the effects of different forms of L. acidophilus on UC, and its mechanism involved in the mitophagy/NLRP3 inflammasome pathway has not been reported. In this study, we found that compared with the heat-killed L. acidophilus and the culture supernatant of L. acidophilus, the live L. acidophilus (La) has the optimal therapeutic effect on UC rats. Furthermore, La evidently increased the contents of SCFAs, inhibited NLRP3 inflammasome and facilitated autophagy. SCFAs regulated by La balanced inflammation homeostasis and improved intestinal barrier dysfunctions in vitro and in vivo, which was achieved by activating the mitophagy/NLRP3 inflammasome pathway. Moreover, PCR analysis indicated that the aforementioned effects of SCFAs regulated by La may be due to the activation of G protein-coupled receptors. These findings provided guidance for the application of L. acidophilus in daily life and provided a new molecular target for interactions among L. acidophilus, its metabolites and host immunity.