Bacterial resistance caused by β-lactamases has been a major threat to public health around the world, seriously weakening the efficacy of β-lactam antibiotics, the most widely used therapeutic agents against infectious diseases. To detect the bacterial resistance to β-lactam antibiotics, particularly specific type of β-lactam antibiotics, in a rapid manner, we report herein a relay-response chemiluminescence assay. This assay mainly consists of two reagents: a β-lactam-caged thiophenol and a thiophenol-sensitive chemiluminescence reporter, both of which are synthetically feasible. The selective hydrolysis of β-lactam by β-lactamase leads to the releasing of free thiophenol, which then triggers the emission of a chemiluminescence signal in a relay manner. Three thiophenol-caged β-lactams, structural analogues of cephalothin, cefotaxime, and meropenem, respectively, have been synthesized. And the application of this assay with these analogues of β-lactam antibiotics allows fast detection of β-lactamase-expressing resistant bacteria and, more impressively, provides detailed information on the resistant scope of bacteria.
Abstract Chronic spontaneous urticaria (CSU) comes with gut dysbiosis, but its relevance remains elusive. Here we use metagenomics sequencing and short-chain fatty acids metabolomics and assess the effects of human CSU fecal microbial transplantation, Klebsiella pneumoniae , Roseburia hominis , and metabolites in vivo. CSU gut microbiota displays low diversity and short-chain fatty acids production, but high gut Klebsiella pneumoniae levels, negatively correlates with blood short-chain fatty acids levels and links to high disease activity. Blood lipopolysaccharide levels are elevated, link to rapid disease relapse, and high gut levels of conditional pathogenic bacteria. CSU microbiome transfer and Klebsiella pneumoniae transplantation facilitate IgE-mediated mast cell(MC)-driven skin inflammatory responses and increase intestinal permeability and blood lipopolysaccharide accumulation in recipient mice. Transplantation of Roseburia hominis and caproate administration protect recipient mice from MC-driven skin inflammation. Here, we show gut microbiome alterations, in CSU, may reduce short-chain fatty acids and increase lipopolysaccharide levels, respectively, and facilitate MC-driven skin inflammation.
Cutaneous squamous cell carcinoma is prone to metastasis at its advanced stage. Previous studies reported that the ubiquitin-conjugating enzyme E2C is a proto-oncogene that causes cancer in various tumors. This study aimed to investigate the role of the ubiquitin-conjugating enzyme E2C in cutaneous squamous cell carcinoma. We initially analyzed the expression profile in the gene expression omnibus database. We obtained the intersection genes, established a protein-protein interaction network, and identified ubiquitin-conjugating enzyme E2C as the key gene. Furthermore, microarray data revealed that the ubiquitin-conjugating enzyme E2C expression is elevated in cutaneous squamous cell carcinoma. Ubiquitin-conjugating enzyme E2C overexpression plasmids and siRNA of ubiquitin-conjugating enzyme E2C were transfected into cutaneous squamous cell carcinoma cells to analyze the effects of ubiquitin-conjugating enzyme E2C on cutaneous squamous cell carcinoma cell proliferation, migration, and invasion. The expression of associated proteins was detected using Western blot assay to confirm the impact of the ubiquitin-conjugating enzyme E2C on the phosphatidylinositol-3-kinase/threonine kinase signaling pathway. Ubiquitin-conjugating enzyme E2C knockdown inhibited cutaneous squamous cell carcinoma cell viability, migration, and invasion, and the phosphatidylinositol-3-kinase/threonine kinase signaling pathway, while ubiquitin-conjugating enzyme E2C overexpression had opposite results. In summary, Ubiquitin-conjugating enzyme E2C is a potential therapeutic target for cutaneous squamous cell carcinoma since it encourages the proliferation and migration of cutaneous squamous cell carcinoma cells.
A fluorogenic probe for the specific detection of IMP-1 β-lactamase activity has been developed. This imaging reagent features a unique trans-acetylamino cephalosporin as an enzymatic recognition moiety, exhibiting excellent selectivity to IMP-1 β-lactamase over other β-lactamases, including serine- and metallo-β-lactamases. The selective activation of the probe by IMP-1 β-lactamase leads to over 30-fold enhancement in the fluorescence intensity, which allows enzyme activity to be reported with high sensitivity.
Chronic spontaneous urticaria (CSU) is defined as recurrent episodes of spontaneous wheal development and/or angioedema for more than six weeks and at least twice a week. The core link in the pathogenesis of CSU is the activation of mast cells, T cells, eosinophils, and other immune cells infiltrating around the small venules of the lesion. Increased vascular permeability, vasodilatation, and recruitment of inflammatory cells directly depend on mast cell mediators’ release. Complex regulatory systems tightly influence the critical roles of mast cells in the local microenvironment. The bias toward Th2 inflammation and autoantibodies derived from B cells, histamine expressed by basophils, and initiation of the extrinsic coagulation pathway by eosinophils or monocytes exerts powerful modulatory influences on mast cells. Cell-to-cell interactions between mast cells and eosinophils/T cells also are regulators of their function and may involve CSU’s pathomechanism. This review summarizes up-to-date knowledge regarding the crosstalk between mast cells and other immune cells, providing the impetus to develop new research concepts and treatment strategies for CSU.
Chronic spontaneous urticaria (CSU) is defined as the recurrent episodes of spontaneous wheals and/or angioedema for more than 6 weeks, and at least twice a week. The core link in the pathogenesis of CSU is the activation of mast cells and other immune cells caused by various reasons. These activated immune cells release a series of inflammatory active mediators such as histamine, arachidonic acid metabolites, chemokines, etc., resulting in clinical features such as wheals and/or edema. However, the specific mechanisms leading to the activation of various immune cells have not been fully elucidated. Previous evidence has shown that about 50% of CSU patients have potential autoimmune reactions. Here we review the functional implication of immune cells in CSU focusing on the crosstalk between them and discuss whether their crosstalk promotes the occurrence and development of the disease.
Chronic spontaneous urticaria (CSU) is a common and recurrent autoimmune-related disease with unclear pathogenesis. Dysfunction of immune cells, such as T cells, mast cells, and basophils, is involved. Bacillus Calmette-Guerin polysaccharide nucleic acid (BCG-PSN), an immunomodulator partially extracted from BCG, can be used in the combined treatment of CSU with an unknown mechanism.To study the therapeutic effect and mechanism of BCG-PSN on CSU, we initially assessed the clinical efficacy in 110 enrolled CSU patients of 4-week antihistamine monotherapy vs. antihistamine plus BCG-PSN combined therapy. Subsequently, to explore the further mechanism of BCG-PSN, the mast cell line RBL-2H3 pretreated with BCG-PSN was used to evaluate the transcriptional expression profiles via lncRNA sequencing. Real time PCR was conducted to validate the candidate gene expression.We found no significant difference in treatment efficacy between the BCG-PSN group (71.7%) and the monotherapy group (71.9%). However, the average time of complete relief in the BCG-PSN group was significantly shorter than that in the monotherapy group (36.77 ± 17.33 vs. 51.27 ± 16.80, p = 0.026). In vitro experiments showed that BCG-PSN inhibited β-hexosaminidase release rates in IgE-sensitized RBL-2H3 cells (p < 0.001). Sequencing data revealed the expression profiles of functional genes, including a significant decrease in Erb-B2 receptor tyrosine kinase 4, which can be regulated by the nuclear factor kappa B (NF-κB) pathway.CSU is a chronic, recurrent disease with complex pathogenesis. Mast cells and basophils are the primary target cells of the disease. BCG-PSN decrease the β-HEX release rates and regulated IgE-mediated mast cell activation in RBL-2H3 cells by mediating immune-related gene expression including ERBB4. These findings suggest that BCG-PSN may mediate ERBB4 expression via the NF-κB pathway and may have value in the treatment of CSU.
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