Time-Resolved Atomistic Imaging and Statistical Analysis of Daptomycin Oligomers with and without Calcium Ions.

Daptomycin (DP) is effective against multiple drug-resistant Gram-positive pathogens because of its distinct mechanism of action. An accepted mechanism includes Ca2+-triggered aggregation of the DP molecule to form oligomers. DP and its oligomers have so far defied structural analysis at a molecular level. We studied the ability of DP molecule to aggregate by itself in water, the effects of Ca2+ ions to promote the aggregation, and the connectivity of the DP molecules in the oligomers by the combined use of dynamic light scattering in water and atomic-resolution cinematographic imaging of DP molecules captured on a carbon nanotube on which the DP molecule is installed as a fishhook. We found that the DP molecule aggregates weakly into dimers, trimers, and tetramers in water, and strongly in the presence of calcium ions, and that the tetramer is the largest oligomer in homogeneous aqueous solution. The dimer remains as the major species, and we propose a face-to-face stacked structure based on dynamic imaging using millisecond and angstrom resolution transmission electron microscopy. The tetramer in its cyclic form is the largest oligomer observed, while the trimer forms in its linear form. The study has shown that the DP molecule has an intrinsic property of forming tetramers in water, which is enhanced by the presence of calcium ions. Such experimental structural information will serve as a platform for future drug design. The data also illustrate the utility of cinematographic recording for the study of self-organization processes.

Three-port single-intercostal versus uniportal thoracoscopic segmentectomy for the treatment of lung cancer: a propensity score matching analysis.

BACKGROUND: The purpose of this retrospective study was to demonstrate the safety and feasibility of three-port single-intercostal video-assisted thoracoscopic surgery (SIC-VATS) segmentectomy compared to uniportal video-assisted thoracoscopic surgery (UVATS) segmentectomy. METHODS: We included 544 patients diagnosed with cT1N0M0 non-small-cell lung cancer (NSCLC) who underwent thoracoscopic segmentectomy between January 2020 and August 2021, including 147 and 397 patients who underwent three-port SIC-VATS and UVATS, respectively. After incorporating preoperative clinical variables, we compared surgical outcomes and perioperative indicators between the two groups by propensity score matching analysis. RESULTS: After 1:1 propensity score matching, each group comprised 143 patients with no significant differences in baseline demographics and characteristics. There was no significant difference in operative time (p = 0.469), blood loss (p = 0.501), number of dissected lymph nodes (p = 0.228), dwell time of the main chest drain (p = 0.065), hospital stay (p = 0.243), or major complication rate (p = 0.295) between the three-port SIC-VATS and UVATS groups. CONCLUSIONS: The three-port SIC-VATS was as safe and feasible as UVATS for patients who are diagnosed with early-stage NSCLC.

Neurons in rat orbitofrontal cortex and medial prefrontal cortex exhibit distinct responses in reward and strategy-update in a risk-based decision-making task.

The orbitofrontal cortex (OFC) and the medial prefrontal cortex (mPFC) are known to participate in risk-based decision-making. However, whether neuronal activities of these two brain regions play similar or differential roles during different stages of risk-based decision-making process remains unknown. Here we conducted multi-channel in vivo recordings in the OFC and mPFC simultaneously when rats were performing a gambling task. Rats were trained to update strategy as the task was shifted in two stages. Behavioral testing suggests that rats exhibited different risk preferences and response latencies to food rewards during stage-1 and stage-2. Indeed, the firing patterns and numbers of non-specific neurons and nosepoking-predicting neurons were similar in OFC and mPFC. However, there were no reward-expecting neurons and significantly more reward-excitatory neurons (fired as rats received rewards) in the mPFC. Further analyses suggested that nosepoking-predicting neurons may encode the overall value of reward and strategy, whereas reward-expecting neurons show more intensive firing to a big food reward in the OFC. Nosepoking-predicting neurons in mPFC showed no correlation with decision-making strategy updating, whereas the response of reward-excitatory neurons in mPFC, which were barely observed in OFC, were inhibited during nosepoking, but were enhanced in the post-nosepoking period. These findings indicate that neurons in the OFC and mPFC exhibit distinct responses in decision-making process during reward consumption and strategy updating. Specifically, OFC encodes the overall value of a choice and is thus important for learning and strategy updating, whereas mPFC plays a key role in monitoring and execution of a strategy.

Lactobacillus paracasei modulates LPS-induced inflammatory cytokine release by monocyte-macrophages via the up-regulation of negative regulators of NF-kappaB signaling in a TLR2-dependent manner.

The application of the probiotic lactobacillus is suggested in the treatment of some inflammatory diseases of intestines due to its potential ability to attenuate inflammation. However, the mechanism is not completely understood. In PBMCs, Lactobacillus paracasei (L. Paracasei) down-regulated the LPS-induced production of TNF-α and IL-6. Using a macrophage-like differentiated THP-1 cell line induced by PMA, we investigated the effect of L. paracasei on the production of pro-inflammatory cytokines by monocyte-macrophages. Treatment of the differentiated THP-1 cells with L. paracasei either concurrently with or before LPS challenge attenuated the LPS-induced secretion of TNF-α and IL-1ß. This effect was due to a decrease in IκB phosphorylation and NF-κB nuclear translocation. Furthermore, treatment of the differentiated THP-1 cells with L. paracasei induced the expression of negative regulators of the NF-κB signaling pathway, including the deubiquitinating enzyme A20, suppressor of cytokine signaling (SOCS) 1, SOCS3, and IL-1 receptor-associated kinase (IRAK) 3. Pretreatment with an IRAK4 inhibitor suppressed the L. paracasei-induced expression of these negative regulators and further increased the LPS-mediated expressions of TNF-α and IL-1ß. Moreover, treatment with an antibody against Toll-like receptor (TLR) 2 reversed the effect of L. paracasei on inducing negative regulators and inhibiting TNF-α and IL-1ß productions. Our findings suggest that L. paracasei inhibits the production of pro-inflammatory cytokines by monocyte-macrophages via the induction of negative regulators of the NF-κB signaling pathway in a TLR2-IRAK4-dependent manner.

IL-29 Enhances LPS/TLR4-Mediated Inflammation in Rheumatoid Arthritis.

BACKGROUND/AIMS: Interleukin-29 (IL-29), a critical member of type III interferons (IFNs) family, has been implicated in protecting against viral infection and modulating autoimmune inflammation. Toll-like receptor 4 (TLR4) plays a crucial role in synovial inflammation and may contribute to the pathogenesis of rheumatology arthritis (RA). However, little is known about the modifying effect of IL-29 on TLR4-mediated inflammation in RA. We aim to investigate the potential association between IL-29 and TLR4 in RA. METHODS: Peripheral blood mononuclear cells (PBMCs) and serum from 77 patients with RA and 70 controls were collected to determine levels of IL-29 and TLR4 mRNA by real-time polymerase chain reaction (PCR). Levels of IL-29 and TLR4 in synovial tissues and fluid from 25 RA patients and 24 controls were detected by enzyme-linked immunosorbent assay (ELISA) or western blot assay, respectively. RAW264.7 cells were stimulated by lipopolysaccharide (LPS) and/or IL-29. The production of inflammatory cytokines including IL-6, IL-8 as well as TNF-α and the activation of nuclear factor-κB (NF-κB) signaling were determined. RESULTS: In comparison with controls, increased IL-29 was observed in PBMCs, synovial tissue, serum and synovial fluid of patients with RA. Besides, TLR4 was significantly elevated in PBMCs and synovium of RA patients. Moreover, IL-29 was positively associated with TLR4 in RA, suggested by Pearson's correlation analysis. When RAW264.7 cells were stimulated by LPS with or without IL-29 in vitro, IL-29 could enhance LPS-mediated TLR4 expression and the production of IL-6, IL-8 and TNF-α in RAW264.7 cells via the activation of NF-κB signaling. CONCLUSION: The present study suggests, for the first time, that IL-29 can aggravate LPS/TLR4-mediated inflammation in RA depending on NF-κB signaling activation.

Stabilizing NASICON-type Na4MnCr(PO4)3 by Ti-substitution toward a high-voltage cathode material for sodium ion batteries.

The sodium superionic conductor Na4MnCr(PO4)3 gains increasing attention owing to its three-dimensional structure and the three-electron reaction. However, rapid structure degradation during cycling is the major challenge for its practical application. Herein, Ti4+ is utilized to replace a portion of Mn2+ in Na4MnCr(PO4)3. The low redox voltage and d0 electronic configuration of the Ti4+ ions are helpful to suppress the structure alteration and improve electronic conduction. Consequently, the as-prepared Na3.4Mn0.7Ti0.3Cr(PO4)3/C cathode exhibits a remarkable good 91.0% capacity retention after 500 cycles at 10C rate, with exceptional rate capacities of 99.5 mAh g-1 and 81.0 mAh g-1 at 5C and 10C rate, respectively. Furthermore, based on ≈2.86-electron reactions involving Mn2+/Mn3+ (3.5 V), Mn3+/Mn4+ (4.1 V), Cr3+/Cr4+ (4.3 V), and Ti3+/Ti4+ (2.1 V), the material can provide an energy density of approximately 541.6 Wh kg-1, slightly surpassing that of Na4MnCr(PO4)3. Ex-situ XRD investigation further elucidates that throughout the entire charge-discharge process, the Ti-substituted material experiences highly reversible solid-solution and two-phase reactions. Additionally, Ti substitution can greatly promote the interfacial charge transfer of the material and suppress the decomposition of the electrolyte during cycling. This work might open a new insight for designing sodium-ion battery cathode materials with good cycling stability and high energy density.

Sustainable synthesis of Ni, Mn co-doped FePO4@C cathode material for Na-ion batteries.

Olivine FePO4 is widely regarded as an optimal cathode material for sodium-ion batteries due to its impressive theoretical capacity of 177.7 mAh g-1. Nonetheless, the material's limited application stems from its intrinsic low electronic and ionic conductivities and ion diffusion rate. Previously, most modifications of olivine FePO4 are conducted through electrochemical or ion exchange processes in organic solvents, which severely restricted its potential for large-scale applications. In this research, a novel water-based ion exchange method is proposed for the synthesis of Ni-doped, Mn-doped, and Ni, Mn co-doped FePO4@C, which is non-toxic, cost-effective, and demonstrating promising prospects for various applications. Fe2.7Mn0.2Ni0.1PO4@C (0.2Mn0.1Ni-FP@C) is synthesized by a straightforward ion exchange method in aqueous media. The material exhibits a discharge capacity of 154.4 mAh g-1 at 0.1C rate. After 300 cycles at 1C, the capacity retention rate remains at 70.7 %. Numerous tests and calculations conducted in this study demonstrate that 0.2Mn0.1Ni-FP@C, modified through Mn3+ and Ni3+ co-doping, exhibits superior electrochemical performance due to its enhanced electronic conductivity and ion diffusion rate.

LOX and Its Methylation Impact Prognosis of Diseases and Correlate with TAM Infiltration in ESCA.

Background: ESCA is one of the digestive tract tumors with a high fatality. It is implicated in an intricate gene regulation process, but the pathogenesis remains ambiguous. Methods: The study used the packages of Limma from R software to analyze DEGs of ESCA in the GEO database and TCGA database. We employed the DAVID website for enrichment analysis, and the string database constructed the PPI network. Hub genes were identified from ESCA DEGs with Cytoscape MCODE. We evaluated the clinical relevance of LOX expression and its DNA methylation in the cBioPortal database and explored the roles of LOX in ESCA immunity, especially immune cell infiltration levels and immune checkpoint expression, by immunedeconv package of R software. Conclusions: The overexpression of LOX in ESCA is regulated by DNA hypomethylation; LOX overexpression or LOX hypomethylation can predict a worse prognosis in patients with ESCA. Besides, LOX may be involved in TIME regulation, promoting the infiltration levels and function of TAM. Hence, high LOX expression affected by DNA hypomethylation has an essential role in patients with ESCA, which may become an effective prognostic marker and therapeutic target.

Changes of inflammation-associated cytokine expressions during early phase of experimental endotoxic shock in macaques.

AIM: To study changes of inflammation-associated cytokine expressions during early phase of endotoxic shock in macaque. METHODS: Experiments were performed in Macaque mulatta treated with LPS 2.8 mg/kg in shock model group or with normal saline in control group. Blood samples were collected before, or 60 min, or 120 min after LPS injection, respectively. Liver and spleen tissues were obtained at 120 min after LPS injection. The plasma levels of TNF-alpha, IL-1beta, IL-10 and IL-12P40 were determined by double-antibody sandwich ELISA with antibodies against human cytokines. The mRNA levels of TNF-alpha, IL-1beta, and IL-18 in peripheral blood mononuclear cells (PBMCs), liver and spleen were examined by real-time fluorescence semi-quantitative RT-PCR with the primers based on human genes. RESULTS: Mean systemic arterial pressure (MAP), systemic vascular resistance index (SVRI) and left ventricular work index (LVWI) of macaques were significant declined in shock model group on average 60 min after LPS injection. The plasma levels of TNF-alpha and IL-10 were significantly increased 60 min after LPS injection and then decreased. The plasma levels of IL-1beta and IL-12P40 were significantly increased at 120 min after LPS injection. The mRNA levels of TNF-alpha and IL-1beta were significantly increased 60 min after LPS stimulation in PBMCs and 120 min after LPS stimulation in livers. The mRNA level of IL-18 was significantly increased 120 min after LPS stimulation in PBMCs and livers. But in spleen, only TNF-alpha mRNA level in LPS group was significantly higher 120 min after LPS stimulation, compared with that in control group. CONCLUSION: An endotoxic shock model of Macaque mulatta was successfully established. Both antibodies for ELISA and PCR primers based on human cytokine assays were successfully applied to detect macaque cytokines. In the model, inflammatory cytokines, such as TNF-alpha, IL-1beta, IL-12 and IL-18 as well as anti-inflammation cytokine IL-10, were released at very early phase of endotoxic shock within 120 min after LPS injection. PBMCs and liver cells might be the important sources of these cytokines.

Lactobacillus isolates from healthy volunteers exert immunomodulatory effects on activated peripheral blood mononuclear cells.

As probiotics in the gut, Lactobacilli are believed to play important roles in the development and maintenance of both the mucosal and systemic immune system of the host. This study was aimed to investigate the immuno-modulatory function of candiate lactobacilli on T cells. Lactobacilli were isolated from healthy human feces and the microbiological characteristics were identified by API 50 CHL and randomly amplified polymorphic DNA (RAPD) assays. Anti-CD3 antibody activated peripheral blood mononuclear cells (PBMCs) were treated by viable, heat-killed lactobacilli and genomic DNA of lactobacilli, and cytokine profiles were tested by ELISA. Isolated lactobacilli C44 and C48 were identified as L. acidophilus and L. paracacei, which have properties of acid and bile tolerance and inhibitor effects on pathogens. Viable and heat-killed C44 and C48 induced low levels of proinflammatory cytokines (TNF-α, IL-6 and IL-8) and high levels of IFN-γ and IL-12p70 in PBMCs. In anti-CD3 antibody activated PBMCs, viable and heat-killed C44 increased Th2 cytokine levels (IL-5, IL-6 and IL-10), and simultaneously enhanced Th1 responses by inducing IFN-γ and IL-12p70 production. Different from that of lactabacillus strains, their genomic DNA induced low levels of IL-12p70, IFN-γ and proinflammatory cytokines in PBMCs with or without anti-CD3 antibody activation. These results provided in vitro evidence that the genomic DNA of strains of C44 and C48, especially C44, induced weaker inflammation, and may be potentially applied for treating allergic diseases.

Gene profile of chemokines on hepatic stellate cells of schistosome-infected mice and antifibrotic roles of CXCL9/10 on liver non-parenchymal cells.

Hepatic stellate cells (HSCs) play a key role in the development of liver fibrosis caused by schistosomiasis. Chemokines were widely expressed and involved in cellular activation, proliferation and migration in inflammatory and infectious diseases. However, little is known about the expressions of chemokines on HSCs in the schistosoma infection. In addition, the roles of chemokines in pathogenesis of liver fibrosis are not totally clear. In our study, we used microarray to analyze the temporal gene expressions of primary HSCs isolated from mice with both acute and chronic schistosomiasis. Our microarray data showed that most of the chemokines expressed on HSCs were upregulated at 3 weeks post-infection (p.i) when the egg granulomatous response was not obviously evoked in the liver. However, some of them like CXCL9, CXCL10 and CXCL11 were subsequently decreased at 6 weeks p.i when the granulomatous response reached the peak. In the chronic stage, most of the differentially expressed chemokines maintained persistent high-abundances. Furthermore, several chemokines including CCR2, CCR5, CCR7, CXCR3, CXCR4, CCL2, CCL5, CCL21, CXCL9 and CXCL10 were expressed by HCSs and the abundances of them were changed following the praziquantel treatment in the chronic stage, indicating that chemokines were possibly necessary for the persistence of the chronic stage. In vitro experiments, hepatic non-parenchymal cells, primary HSCs and human HSCs line LX-2 were stimulated by chemokines. The results showed that CXCL9 and CXCL10, but not CXCL11 or CXCL4, significantly inhibited the gene expressions of Col1α1, Col3α1 and α-SMA, indicating the potential anti-fibrosis effect of CXCL9 and CXCL10 in schistosomiasis. More interestingly, soluble egg antigen (SEA) of Schistosoma japonicum was able to inhibit transcriptional expressions of some chemokines by LX-2 cells, suggesting that SEA was capable of regulating the expression pattern of chemokine family and modulating the hepatic immune microenvironment in schistosomiasis.

New insight into the antifibrotic effects of praziquantel on mice in infection with Schistosoma japonicum.

BACKGROUND: Schistosomiasis is a parasitic disease infecting more than 200 million people in the world. Although chemotherapy targeting on killing schistosomes is one of the main strategies in the disease control, there are few effective ways of dealing with liver fibrosis caused by the parasite infection in the chronic and advanced stages of schistosomiasis. For this reason, new strategies and prospective drugs, which exert antifibrotic effects, are urgently required. METHODS AND FINDINGS: The antifibrotic effects of praziquantel were assessed in the murine models of schistosomiasis japonica. Murine fibrosis models were established by cutaneous infection with 14 ± 2 Schistosoma japonicum cercariae. Then, the mice of both chronic (8 weeks post-infection) and advanced (15 weeks post-infection) schistosomiasis were treated by gavage of praziquantel (250 mg/kg, once daily for 3 days) to eliminate worms, and followed by praziquantel anti-fibrosis treatment (300 mg/kg, twice daily for 30 days). The fibrosis-related parameters assessed were areas of collagen deposition, content of hydroxyproline and mRNA expressions of Col1α1, Col3α1, α-SMA, TGF-ß, MMP9, TIMP1, IL-4, IL-10, IL-13 and IFN-γ of liver. Spleen weight index, alanine aminotransferase activity and liver portal venous pressure were also measured. The results showed that anti-fibrosis treatment improved liver fibrosis, splenomegaly, hepatic function, as well as liver portal hypertension. In order to confirm the anti-fibrotic properties of praziquantel, we established a CCL(4)-induced model and revealed that CCL(4)-induced liver fibrosis was inhibited by PZQ treatment for 30 days. Furthermore, we analyzed the effects of praziquantel on mouse primary hepatic stellate cells (HSCs). It is indicated that mRNA expressions of Col1α1, Col3α1, α-SMA, TGF-ß, MMP9 and TIMP1 of HSCs were all inhibited after praziquantel anti-parasite treatments. CONCLUSIONS: The significant amelioration of hepatic fibrosis by praziquantel treatment validates it as a promising drug of anti-fibrosis and offers potential of a new chemotherapy for hepatic fibrosis resulting from schistosomiasis.

The abnormal apoptosis of T cell subsets and possible involvement of IL-10 in systemic lupus erythematosus.

To study the apoptosis of lymphocyte subpopulations in systemic lupus erythematosus (SLE) patients and the possible role of IL-10 in this apoptosis involved in the pathogenesis of SLE, three color fluorescence and flow cytometry were used to investigate the early apoptosis of lymphocyte subsets from freshly separated or cultured peripheral blood mononuclear cells (PBMCs). ELISA was employed to detect the levels of IL-10 in serum and the levels of sFas and sFasL in cultured PBMC supernatants, and the results of sFas and sFasL were confirmed by real-time PCR of Fas and FasL mRNA. The results showed that in cells from SLE patients, the apoptosis of CD3+, CD4+, and CD8+ T cells was distinctly increased, and the percentage of CD4+ cells and the CD4/CD8 ratio was significantly decreased, as compared with normal controls. The apoptosis of T lymphocytes cultured with SLE serum was markedly higher than that of cells cultured with control's serum. Blockade of interleukin-10 (IL-10) activation by an anti-IL-10 antibody reduced the SLE serum induced apoptosis of CD4+ and CD8+ T cells. The levels of sFas and sFasL in the culture supernatant and Fas and FasL mRNA expressions in cultured cells were significantly higher in the SLE serum-cultured groups, but decreased evidently in the presence of the anti-IL-10 antibody. Above findings suggested that SLE cells showed abnormally high apoptosis of T lymphocytes, especially of the CD4+ subpopulation, resulting in a decreased CD4/CD8 ratio. The high percentage of apoptotic T cells in SLE patients may be related to the high levels of IL-10 in SLE serum, as IL-10 may induce the abnormally activated T cells to trigger apoptosis via the Fas-FasL pathway.

A novel NASICON-type Na3MnTi0.5Zr0.5(PO4)3 cathode material with multivalent redox reaction for high performance sodium-ion batteries.

Na3MnZr(PO4)3, a typical manganese-based NASICON-type material, has consistently been at the forefront of research on cathode materials for sodium-ion batteries due to the abundant manganese reserve and high operating voltage. However, the severe Jahn-Teller effect, poor electronic conductivity and kinetic limitation of Na3MnZr(PO4)3 impose constraints on its rate capability and cycling performance, thereby hindering its practical application. To address this challenge, a ternary NASICON-type material Na3MnTi0.5Zr0.5(PO4)3/C, with a multi-metal synergistic effect, is proposed in this study. The substitution of Ti at Zr site significantly mitigates the Jahn-Teller effect induced by Mn3+. Furthermore, the stability of the ZrO bond is enhanced, leading to a more robust crystal structure overall. Cyclic voltammetry and constant-current intermittent titration techniques reveal that the appropriate Ti substitution markedly boosts the electronic conductivity and Na+ diffusion coefficient of the electrode material, thereby mitigating polarization effects and expediting electrode reaction rates. Leveraging the multi-effect of Ti substitution, the prepared Na3MnTi0.5Zr0.5(PO4)3/C presents an improved electrochemical performance. Notably, Na3MnTi0.5Zr0.5(PO4)3/C enables a high discharge capacity of 71.0 mAh g-1 at 10C and maintains 78.8 % capacity after 1000 cycles at 2C rate. This investigation establishes a robust theoretical foundation for comprehending the synergistic effects of multimetal systems in NASICON materials and offers insights into the development of cost-effective, high-performance cathode materials.