L-arginine attenuates Streptococcus uberis-induced inflammation by decreasing miR155 level.

L-arginine, as an essential substance of the immune system, plays a vital role in innate immunity. MiR155, a multi-functional microRNA, has gained importance as a regulator of homeostasis in immune cells. However, the immunoregulatory mechanism between L-arginine and miR155 in bacterial infections is unknown. Here, we investigated the potential role of miR155 in inflammation and the molecular regulatory mechanisms of L-arginine in Streptococcus uberis (S. uberis) infections. And we observed that miR155 was up-regulated after infection, accompanying the depletion of L-arginine, leading to metabolic disorders of amino acids and severe tissue damage. Mechanically, the upregulated miR155 mediated by the p65 protein played a pro-inflammatory role by suppressing the suppressor of cytokine signaling 6 (SOCS6)-mediated p65 ubiquitination and degradation. This culminated in a violently inflammatory response and tissue damage. Interestingly, a significant anti-inflammatory effect was revealed in L-arginine supplementation by reducing miR155 production via inhibiting p65. This work firstly uncovers the pro-inflammatory role of miR155 and an anti-inflammatory mechanism of L-arginine in S.uberis infection with a mouse mastitis model. Collectively, we provide new insights and strategies for the prevention and control of this important pathogen, which is of great significance for ensuring human food health and safety.

Functional diversity of voltage-gated sodium channel in Drosophila suzukii (Matsumura).

BACKGROUND: The larvae of Drosophila suzukii Matsumura feed directly inside the fruit, causing catastrophic damage to orchards. The misuse of pyrethroid insecticides during the control period has led to increasing resistance of D. suzukii to pyrethroids acting on the voltage-gated sodium channel (VGSC). RESULTS: In this study, the sodium channel of D. suzukii was cloned (DsNav 5 GenBank number: OQ871532). The results of multiple-sequence alignment showed that the homology of sodium channel between D. suzukii and Drosophila melanogaster was as high as 95.3%. Analysis of transcripts from 62 variants of D. suzukii VGSC revealed a total of six alternative splicing sites (exons u, j, a, b, e, and h) and 33 RNA editing. Exons j, a, b, e, and h are conserved in D. melanogaster and other insects, whereas exon u has never been reported before. The number of A-to-I was distinctly more than that of U-to-C for RNA editing. All D. suzukii VGSC variants were expressed in Xenopus oocytes, but only one (type 5) was able to produce robust currents and nine produce weak currents. DsNav 5 with TipE of D. melanogaster co-expresses current better than its own TipE. Subsequently, tetrodotoxin was verified to be a blocker of VGSC, and the gating properties of DsNav 5 were investigated. CONCLUSION: These findings proved that the VGSC of D. suzukii has not only the basic gating properties, but also the diversity of gating properties. This study also laid a foundation for the study of pyrethroid resistance mechanism of VGSC in D. suzukii. © 2023 Society of Chemical Industry.

PFKFB3-Meditated Glycolysis via the Reactive Oxygen Species-Hypoxic Inducible Factor 1α Axis Contributes to Inflammation and Proliferation of Staphylococcus aureus in Epithelial Cells.

Mastitis caused by antibiotic-resistant strains of Staphylococcus aureus is a significant concern in the livestock industry due to the economic losses it incurs. Regulating immunometabolism has emerged as a promising approach for preventing bacterial inflammation. To investigate the possibility of alleviating inflammation caused by S aureus infection by regulating host glycolysis, we subjected the murine mammary epithelial cell line (EpH4-Ev) to S aureus challenge. Our study revealed that S aureus can colonize EpH4-Ev cells and promote inflammation through hypoxic inducible factor 1α (HIF1α)-driven glycolysis. Notably, the activation of HIF1α was found to be dependent on the production of reactive oxygen species (ROS). By inhibiting PFKFB3, a key regulator in the host glycolytic pathway, we successfully modulated HIF1α-triggered metabolic reprogramming by reducing ROS production in S aureus-induced mastitis. Our findings suggest that there is a high potential for the development of novel anti-inflammatory therapies that safely inhibit the glycolytic rate-limiting enzyme PFKFB3.

Research related to the diagnosis of prostate cancer based on machine learning medical images: A review.

BACKGROUND: Prostate cancer is currently the second most prevalent cancer among men. Accurate diagnosis of prostate cancer can provide effective treatment for patients and greatly reduce mortality. The current medical imaging tools for screening prostate cancer are mainly MRI, CT and ultrasound. In the past 20 years, these medical imaging methods have made great progress with machine learning, especially the rise of deep learning has led to a wider application of artificial intelligence in the use of image-assisted diagnosis of prostate cancer. METHOD: This review collected medical image processing methods, prostate and prostate cancer on MR images, CT images, and ultrasound images through search engines such as web of science, PubMed, and Google Scholar, including image pre-processing methods, segmentation of prostate gland on medical images, registration between prostate gland on different modal images, detection of prostate cancer lesions on the prostate. CONCLUSION: Through these collated papers, it is found that the current research on the diagnosis and staging of prostate cancer using machine learning and deep learning is in its infancy, and most of the existing studies are on the diagnosis of prostate cancer and classification of lesions, and the accuracy is low, with the best results having an accuracy of less than 0.95. There are fewer studies on staging. The research is mainly focused on MR images and much less on CT images, ultrasound images. DISCUSSION: Machine learning and deep learning combined with medical imaging have a broad application prospect for the diagnosis and staging of prostate cancer, but the research in this area still has more room for development.

Comparative and phylogenetic analyses of nine complete chloroplast genomes of Orchidaceae.

The orchid family has 200,000 species and 700 genera, and it is found worldwide in the tropics and subtropics. In China, there are 1247 species and subspecies of orchids belonging to the Orchidaceae family. Orchidaceae is one of the most diverse plant families in the world, known for their lush look, remarkable ecological tolerance, and capability for reproduction. It has significant decorative and therapeutic value. In terms of evolution, the orchid family is one of the more complicated groups, but up until now, little has been known about its affinities. This study examined the properties of 19 chloroplast (cp) genomes, of which 11 had previously been published and nine had only recently been revealed. Following that, topics such as analysis of selection pressure, codon usage, amino acid frequencies, repeated sequences, and reverse repeat contraction and expansion are covered. The Orchidaceae share similar cp chromosomal characteristics, and we have conducted a preliminary analysis of their evolutionary connections. The cp genome of this family has a typical tepartite structure and a high degree of consistency across species. Platanthera urceolata with more tandem repeats of the cp genome. Similar cp chromosomal traits can be seen in the orchidaceae. Galearis roborowskyi, Neottianthe cucullata, Neottianthe monophylla, Platanthera urceolata and Ponerorchis compacta are the closest cousins, according to phylogenetic study.

Taurine reduction of injury from neutrophil infiltration ameliorates Streptococcus uberis-induced mastitis.

Mastitis is a common disease of dairy cows characterized by infiltration of leukocytes, especially neutrophils, resulting in increased permeability of the blood-milk barrier (BMB). Taurine, a functional nutrient, has been shown to have anti-inflammatory and antioxidant effects. Here, we investigated the regulatory effects and mechanisms of taurine on the complex immune network of the mammary gland in Streptococcus uberis (S. uberis) infection. We found that taurine had no direct effect on CXCL2-mediated neutrophil chemotaxis. However, it inhibited MAPK and NF-κB signalings by modulating the activity of TAK1 downstream of TLR2, thereby reducing CXCL2 expression in macrophages to reduce neutrophil recruitment in S. uberis infection. Further, the AMPK/Nrf2 signaling pathway was activated by taurine to help mitigate oxidative damage, apoptosis and disruption of tight junctions in mammary epithelial cells caused by hypochlorous acid, a strong oxidant produced by neutrophils, thus protecting the integrity of the mammary epithelial barrier. Taurine protects the BMB from damage caused by neutrophils via blocking the macrophage-CXCL2-neutrophil signaling axis and increasing the antioxidant capacity of mammary epithelial cells.

The therapeutic potential for targeting CSE/H2S signaling in macrophages against Escherichia coli infection.

Macrophages play a pivotal role in the inflammatory response to the zoonotic pathogen E. coli, responsible for causing enteric infections. While considerable research has been conducted to comprehend the pathogenesis of this disease, scant attention devoted to host-derived H2S. Herein, we reported that E. coli infection enhanced the expression of CSE in macrophages, accompanied by a significantly increased inflammatory response. This process may be mediated by the involvement of excessive autophagy. Inhibition of AMPK or autophagy with pharmacological inhibitors could alleviate the inflammation. Additionally, cell model showed that the mRNA expression of classic inflammatory factors (Il-1ß, Il-6), macrophage polarization markers (iNOS, Arg1) and ROS production was significantly down-regulated after employing CSE specific inhibitor PAG. And PAG is capable of inhibiting excessive autophagy through the LKB1-AMPK-ULK1 axis. Interestingly, exogenous H2S could suppress inflammation response. Our study emphasizes the importance of CSE in regulating the macrophage-mediated response to E. coli. Increased CSE in macrophages leads to excessive inflammation, which should be considered a new target for drug development to treat intestinal infection.

Classification of Histological Types and Stages in Non-small Cell Lung Cancer Using Radiomic Features Based on CT Images.

Non-invasive diagnostic method based on radiomic features in patients with non-small cell lung cancer (NSCLC) has attracted attention. This study aimed to develop a CT image-based model for both histological typing and clinical staging of patients with NSCLC. A total of 309 NSCLC patients with 537 CT series from The Cancer Imaging Archive (TCIA) database were included in this study. All patients were randomly divided into the training set (247 patients, 425 CT series) and testing set (62 patients, 112 CT series). A total of 107 radiomic features were extracted. Four classifiers including random forest, XGBoost, support vector machine, and logistic regression were used to construct the classification model. The classification model had two output layers: histological type (adenocarcinoma, squamous cell carcinoma, and large cell) and clinical stage (I, II, and III) of NSCLC patients. The area under the receiver operating characteristic curve (AUC), accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) with 95% confidence interval (CI) were utilized to evaluate the performance of the model. Seven features were selected for inclusion in the classification model. The random forest model had the best classification ability compared with other classifiers. The AUC of the RF model for histological typing and clinical staging of NSCLC patients in the testing set was 0.700 (95% CI, 0.641-0.759) and 0.881 (95% CI, 0.842-0.920), respectively. The CT image-based radiomic feature model had good classification ability for both histological typing and clinical staging of patients with NSCLC.

AMPK/Drp1 pathway mediates Streptococcus uberis-Induced mitochondrial dysfunction.

Excessive production of reactive oxygen species (ROS) leads to oxidative stress in host cells and affects the progress of disease. Mitochondria are an important source of ROS and their dysfunction is closely related to ROS production. S. uberis is a common causative agent of mastitis. The expression of key enzymes of the mitochondrial apoptotic pathway is increased in mammary epithelial cells after S. uberis stimulation, while expression of proteins related to mitochondrial function is decreased. Drp1, a key protein associated with mitochondrial function, is activated upon infection. Accompanied by mitochondria-cytosol translocation of Drp1, Fis1 expression is significantly upregulated while Mfn1 expression is downregulated implying that the balance of mitochondrial dynamics is disrupted. This leads to mitochondrial fragmentation, decreased mitochondrial membrane potential, higher levels of mROS and oxidative injury. The AMPK activator AICAR inhibits the increased phosphorylation of Drp1 and the translocation of Drp1 to mitochondria by salvaging mitochondrial function in an AMPK/Drp1 dependent manner, which has a similar effect to Drp1 inhibitor Mdivi-1. These data show that AMPK, as an upstream negative regulator of Drp1, ameliorates mitochondrial dysfunction induced by S. uberis infection.

Molecular Epidemiology and Antimicrobial Resistance of Outbreaks of Klebsiella pneumoniae Clinical Mastitis in Chinese Dairy Farms.

Klebsiella pneumoniae is an opportunistic pathogen that causes serious infections in humans and animals. However, the availability of epidemiological information on clinical mastitis due to K. pneumoniae is limited. To acquire new information regarding K. pneumoniae mastitis, data were mined about K. pneumoniae strains on dairy cattle farms (farms A to H) in 7 Chinese provinces in 2021. Hypermucoviscous strains of K. pneumoniae were obtained by the string test. MICs of antimicrobial agents were determined via the broth microdilution method. Ten antimicrobial resistance genes and virulence genes were identified by PCR. The prevalence of K. pneumoniae was 35.91% (65/181), and 100% of the bacteria were sensitive to enrofloxacin. Nine antimicrobial resistance genes and virulence genes were identified and compared among farms. The hypermucoviscous phenotype was present in 94.44% of isolates from farm B, which may be a function of the rmpA virulence gene. Based on these data, the multidrug-resistant strains SD-14 and HB-21 were chosen and sequenced. Genotypes were assayed for K. pneumoniae isolates from different countries and different hosts using multilocus sequence typing (MLST). Ninety-four sequence types (STs) were found, and 6 STs present a risk for spreading in specific regions. Interestingly, ST43 was observed in bovine isolates for the first time. Our study partially reveals the current distribution characteristics of bovine K. pneumoniae in China and may provide a theoretical basis for the prevention and treatment of bovine K. pneumoniae mastitis. IMPORTANCE K. pneumonia is ubiquitous in nature and infects a wide range of hosts, including animals, and humans. It is one of the leading inducements of clinical mastitis (CM) in dairy cows, a prevalent and costly disease that is predominantly associated with bacterial infection. In general, CM caused by Gram-negative bacteria is more difficult to cure than that associated with Gram-positive pathogens, with an average cost per case of 211.03 U.S. dollars (USD) for Gram-negative bacterial infections compared with 133.73 USD for Gram-positive bacterial CM cases. After Escherichia coli, K. pneumoniae is the second most common Gram-negative cause of bovine CM, but it is the most detrimental in terms of decreased milk yield, discarded milk, treatment costs, death, and culling. In view of the economic implications of K. pneumoniae infection in dairy farming, research into population structure and antibiotic resistance is particularly important.

FABP4-mediated lipid droplet formation in Streptococcus uberis-infected macrophages supports host defence.

Foamy macrophages containing prominent cytoplasmic lipid droplets (LDs) are found in a variety of infectious diseases. However, their role in Streptococcus uberis-induced mastitis is unknown. Herein, we report that S. uberis infection enhances the fatty acid synthesis pathway in macrophages, resulting in a sharp increase in LD levels, accompanied by a significantly enhanced inflammatory response. This process is mediated by the involvement of fatty acid binding protein 4 (FABP4), a subtype of the fatty acid-binding protein family that plays critical roles in metabolism and inflammation. In addition, FABP4 siRNA inhibitor cell models showed that the deposition of LDs decreased, and the mRNA expression of Tnf, Il1b and Il6 was significantly downregulated after gene silencing. As a result, the bacterial load in macrophages increased. Taken together, these data demonstrate that macrophage LD formation is a host-driven component of the immune response to S. uberis. FABP4 contributes to promoting inflammation via LDs, which should be considered a new target for drug development to treat infections.

Taurine inhibits Streptococcus uberis-induced NADPH oxidase-dependent neutrophil extracellular traps via TAK1/MAPK signaling pathways.

Neutrophil extracellular traps (NETs) are produced by neutrophil activation and usually have both anti-infective and pro-damage effects. Streptococcus uberis (S. uberis), one of the common causative organisms of mastitis, can lead to the production of NETs. Taurine, a free amino acid abundant in the organism, has been shown to have immunomodulatory effects. In this study, we investigated the molecular mechanisms of S. uberis-induced NETs formation and the regulatory role of taurine. The results showed that NETs had a disruptive effect on mammary epithelial cells and barriers, but do not significantly inhibit the proliferation of S. uberis. S. uberis induced NADPH oxidase-dependent NETs. TLR2-mediated activation of the MAPK signaling pathway was involved in this process. Taurine could inhibit the activation of MAPK signaling pathway and NADPH oxidase by modulating the activity of TAK1, thereby inhibiting the production of ROS and NETs. The effects of taurine on NADPH oxidase and NETs in S. uberis infection were also demonstrated in vivo. These results suggest that taurine can protect mammary epithelial cells and barriers from damage by reducing S. uberis-induced NETs. These data provide new insights and strategies for the prevention and control of mastitis.

Taurine inhibits necroptosis helps to alleviate inflammatory and injury induced by Klebsiella infection.

Klebsiella infection is widely acknowledged to inflict severe inflammatory damage in bovines. Herein, we demonstrate significant death of EpH4-Ev cells incubated with Klebsiella. And compelling evidence shows that Klebsiella infection increases interactions between the Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) and RIPK3, which promotes phosphorylation of RIPK3 and MLKL to induce necroptosis. However, these changes can be partially reversed by taurine and Nec-1s. Moreover, using taurine and Nec-1s to partially inhibit necroptosis significantly reduce TNF-α, IL-1ß and IL-6 levels and NAGase activity induced by Klebsiella infection. Taken together, taurine partially inhibits necroptosis induced by Klebsiella infection and hence alleviates inflammatory and injury in EpH4-Ev cells.

Reduction of ROS-HIF1α-driven glycolysis by taurine alleviates Streptococcus uberis infection.

Antibiotic-resistant strains of Streptococcus uberis (S. uberis) frequently cause clinical mastitis in dairy cows resulting in enormous economic losses. The regulation of immunometabolism is a promising strategy for controlling this bacterial infection. To investigate whether taurine alleviates S. uberis infection by the regulation of host glycolysis via HIF1α, the murine mammary epithelial cell line (EpH4-Ev) and C57BL/6J mice were challenged with S. uberis. Our data indicate that HIF1α-driven glycolysis promotes inflammation and damage in response to the S. uberis challenge. The activation of HIF1α is dependent on mTOR-mediated ROS production. These results were confirmed in vivo. Taurine, an intracellular metabolite present in most animal tissues, has been shown to effectively modulate HIF1α-triggered metabolic reprogramming and contributes to a reduction of inflammation, which reduces mammary tissue damage and prevents mammary gland dysfunction in S. uberis-induced mastitis. These data provide a novel putative prophylactic and therapeutic strategy for amelioration of dairy cow mastitis and bacterial inflammation.

The complete chloroplast genome of a medical herb, Rheum lhasaense (Polygonaceae), from Qinghai-Tibet Plateau in China.

Rheum lhasaense (Polygonaceae) is one of the genuine medicinal herbs in Qinghai-Tibet Plateau, China. Here we report the first chloroplast (cp) genome of R. lhasaense using Illumina NovaSeq 6000 platform. The length of its complete cp genome is 161,820 bp, containing four sub-regions. A large single copy region (LSC) of 87,086 bp and a small single copy region (SSC) of 12,814 bp are separated by a pair of inverted repeat regions (IRs) of 30,960 bp. The complete cp genome of R. lhasaense contains 130 genes, including 85 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. The overall GC content of the cp genome is 37.4%. The phylogenetic analysis, based on 28 cp genomes, suggested that R. lhasaense is closely related to R. acuminatum and R. pumilum.

Taurine protects blood-milk barrier integrity via limiting inflammatory response in Streptococcus uberis infections.

Streptococcus uberis (S. uberis) is an important causative agent of mastitis, leading to significant economic losses to dairy industry. This research used a mouse mastitis model to investigate the protective effects of taurine on mammary inflammatory response and blood-milk barrier integrity in S. uberis challenge. The results showed that taurine attenuated S. uberis-induced mammary histopathological changes, especially neutrophil infiltration. The S. uberis-induced expression of pro-inflammatory mediators were decreased significantly by taurine. Further, we demonstrated that taurine limited the S. uberis-induced inflammatory responses via inhibiting the activation of NF-κB and MAPK signaling pathways. Inflammation usually disrupts the mammary barrier system. The recovery of claudin-3 and occludin expressions indicated that attenuation of inflammatory response by taurine can protect the integrity of blood-milk barrier in S. uberis infection. Taken together, our results reveal that the development of taurine as an effective prevention and control strategy for S. uberis-induced mastitis.

Fully redundant auxiliary system for gyrotron-based terahertz sources in long-term operation.

The Wuhan National High Magnetic Field Center is incorporating the flat-top pulsed magnetic field (FTPMF) into pulse gyrotrons. It will be the first chance to make a pulse-magnet gyrotron available for generating a long-pulse radiation of 100 ms or above without affecting its high operating frequency and high radiation power. However, unlike continuous wave gyrotrons, pulse gyrotrons in long-term operation have their own challenges, namely, misalignment caused by concussions, much stronger low-frequency electromagnetic interference from the pulse magnet, and inevitable explosion. This article will focus on the difficulties faced by pulse gyrotrons in years of operation, discuss the protection and restoration from failures, and, consequently, propose a fully redundant, explosion-proof, and quickly recoverable auxiliary system for long-term operation of pulse gyrotrons. This system integrates the control unit of traditional pulsed magnets and superconducting magnets so that it can be compatible with any form of gyrotron facilities. Therefore, once the FTPMF or the superconducting magnet is available, the long-pulse radiation will be obtained. Several experimental results, including the most recent explosion, show the reliability of the proposed system.

Taurine Reprograms Mammary-Gland Metabolism and Alleviates Inflammation Induced by Streptococcus uberis in Mice.

Streptococcus uberis (S. uberis) is an important pathogen causing mastitis, which causes continuous inflammation and dysfunction of mammary glands and leads to enormous economic losses. Most research on infection continues to be microbial metabolism-centric, and many overlook the fact that pathogens require energy from host. Mouse is a common animal model for studying bovine mastitis. In this perspective, we uncover metabolic reprogramming during host immune responses is associated with infection-driven inflammation, particularly when caused by intracellular bacteria. Taurine, a metabolic regulator, has been shown to effectively ameliorate metabolic diseases. We evaluated the role of taurine in the metabolic regulation of S. uberis-induced mastitis. Metabolic profiling indicates that S. uberis exposure triggers inflammation and metabolic dysfunction of mammary glands and mammary epithelial cells (the main functional cells in mammary glands). Challenge with S. uberis upregulates glycolysis and oxidative phosphorylation in MECs. Pretreatment with taurine restores metabolic homeostasis, reverses metabolic dysfunction by decrease of lipid, amino acid and especially energy disturbance in the infectious context, and alleviates excessive inflammatory responses. These outcomes depend on taurine-mediated activation of the AMPK-mTOR pathway, which inhibits the over activation of inflammatory responses and alleviates cellular damage. Thus, metabolic homeostasis is essential for reducing inflammation. Metabolic modulation can be used as a prophylactic strategy against mastitis.

Taurine Alleviates Streptococcus uberis-Induced Inflammation by Activating Autophagy in Mammary Epithelial Cells.

Streptococcus uberis infection can cause serious inflammation and damage to mammary epithelial cells and tissues that can be significantly alleviated by taurine. Autophagy plays an important role in regulating immunity and clearing invasive pathogens and may be regulated by taurine. However, the relationships between taurine, autophagy, and S. uberis infection remain unclear. Herein, we demonstrate that taurine augments PTEN activity and inhibits Akt/mTOR signaling, which decreases phosphorylation of ULK1 and ATG13 by mTOR and activates autophagy. Activating autophagy accelerates the degradation of intracellular S. uberis, reduces intracellular bacterial load, inhibits over-activation of the NF-κB pathway, and alleviates the inflammation and damage caused by S. uberis infection. This study increases our understanding of the mechanism through which taurine regulates autophagy and is the first to demonstrate the role of autophagy in S. uberis infected MAC-T cells. Our study also provides a theoretical basis for employing nutritional elements (taurine) to regulate innate immunity and control S. uberis infection. It also provides theoretical support for the development of prophylactic strategies for this important pathogen.

Taurine-Mediated IDOL Contributes to Resolution of Streptococcus uberis Infection.

Metabolic alterations occur in pathogenic infections, but the role of lipid metabolism in the progression of bacterial mastitis is unclear. Cross talk between lipid droplets (LDs) and invading bacteria occurs, and targeting of de novo lipogenesis inhibits pathogen reproduction. In this study, we investigate the role(s) of lipid metabolism in mammary cells during Streptococcus uberis infection. Our results indicate that S. uberis induces the synthesis of fatty acids and production of LDs. Importantly, taurine reduces fatty acid synthesis, the abundance of LDs and the in vitro bacterial load of S. uberis These changes are mediated, at least partly, by the E3 ubiquitin ligase IDOL, which is associated with the degradation of low-density lipoprotein receptors (LDLRs). We have identified a critical role for IDOL-mediated fatty acid synthesis in bacterial infection, and we suggest that taurine may be an effective prophylactic or therapeutic strategy for preventing S. uberis mastitis.