Thiolutin, a selective NLRP3 inflammasome inhibitor, attenuates cyclophosphamide-induced impairment of sperm and fertility in mice.

OBJECTIVE: The activation of the NLRP3 inflammasome has been implicated in male infertility. Our study aimed to investigate the therapeutic role of Thiolutin (THL), an inhibitor of the NLRP3 inflammasome, on oligoasthenospermia (OA) and to elucidate its mechanisms. MATERIALS AND METHODS: Semen from 50 OA and 20 healthy males were analyzed to assess the sperm quality and levels of inflammatory markers. Their correlation was determined using Pearson's correlation coefficient. The BALB/c mice were intraperitoneal injected by cyclophosphamide at 60 mg/kg/day for five days to induce OA, followed by a two-week treatment with THL or L-carnitine. Reproductive organ size and H&E staining were determined to observe the organ and seminiferous tubule morphology. ELISA and western blotting were utilized to measure sex hormone levels, inflammatory markers, and NLRP3 inflammasome levels. Furthermore, male and female mice were co-housed to observe pregnancy success rates. RESULTS: OA patients exhibited a decrease in sperm density and motility compared to healthy individuals, along with elevated levels of IL-1ß, IL-18 and NLRP3 inflammasome. In vivo, THL ameliorated OA-induced atrophy of reproductive organs, hormonal imbalance, and improved sperm density, motility, spermatogenesis and pregnancy success rates with negligible adverse effects on weight or liver-kidney function. THL also demonstrated to be able to inhibit the activation of NLRP3 inflammasome and associated proteins in OA mice. DISCUSSION: THL can improve sperm quality and hormonal balance in OA mice through the inhibition of NLRP3 inflammasome activation. Thus, THL holds promising potential as a therapeutic agent for OA.

Niobium Oxide Anode with Lattice Structure Self-Optimization for High-Power and Nearly Zero-Degeneration Battery Operation.

Li+ insertion-induced structure transformation in crystalline electrodes vitally influence the energy density and cycle life of secondary lithium-ion battery. However, the influence mechanism of structure transformation-induced Li+ migration on the electrochemical performance of micro-crystal materials is still unclear and the strategy to profit from such structure transformation remains exploited. Here, an interesting self-optimization of structure evolution during electrochemical cycling in Nb2 O5 micro-crystal with rich domain boundaries is demonstrated, which greatly improves the charge transfer property and mechanical strength. The lattice rearrangement activates the Li+ diffusion kinetics and hinders the particle crack, thus enabling a nearly zero-degeneration operation after 8000 cycles. Full cell paired with lithium cobalt oxides displays an exceptionally high capacity of 176 mA h g-1 at 8000 mA g-1 and excellent long-term durability at 6000 mA g-1 with 63% capacity retention over 2000 cycles. Interestingly, a unique fingerprint based on the intensity ratio of two X-ray diffraction peaks is successfully extracted as a measure of Nb2 O5 electrochemical performance. The structure self-optimization for fast charge transfer and high mechanical strength exemplifies a new battery electrode design concept and opens up a vast space of strategy to develop high-performance lithium-ion batteries with high energy density and ultra-long cycle life.

Mechanisms and origins of switchable regioselectivity of palladium- and nickel-catalyzed allene hydrosilylation with N-heterocyclic carbene ligands: a theoretical study.

The mechanisms and origins for the Pd- and Ni-catalyzed regioselective hydrosilylation of allene have been investigated by means of density functional theory (DFT) calculations. The free-energy profiles of Pd- and Ni-catalyzed reactions with small and bulky N-heterocyclic carbene (NHC) ligands are calculated to determine the mechanism for regioselectivities. The calculation results show that different metals (Ni vs Pd) lead to regiochemical reversals for the hydrosilylation of allene. The allylsilane is the major product via palladium catalysis with small NHC ligand, while the vinylsilane is the major product via nickel catalysis with bulky NHC ligand. Both electronic and steric factors play a key role in the regioselectivities for the hydrosilylation of allene via Pd and Ni catalysts. The calculation results are in good agreement with observed regioselectivities and could provide insights into the design of new catalysts for the regioselectivity of hydrosilylation reactions.

One-Step Synthesis of Three-Dimensional Na3V2(PO4)3/Carbon Frameworks as Promising Sodium-Ion Battery Cathode.

Sodium-ion batteries (SIBs) are essential for large-scale energy storage attributed to the high abundance of sodium. Polyanion Na3V2(PO4)3 (NVP) is a dominant cathode candidate for SIBs because of its high-voltage and sodium superionic conductor (NASICON) framework. However, the electrochemical performance of NVP is hindered by the inherently poor electronic conductivity, especially for extreme fast charging and long-duration cycling. Herein, we develop a facile one-step in-situ polycondensation method to synthesize the three-dimensional (3D) Na3V2(PO4)3/holey-carbon frameworks (NVP@C) by using melamine as carbon source. In this architecture, NVP crystals intergrown with the 3D holey-carbon frameworks provide rapid transport pathways for ion/electron transmission to increase the ultrahigh rate ability and cycle capability. Consequently, the NVP@C cathode possesses a high reversible capacity of 113.9 mAh g-1 at 100 mA g-1 and delivers an outstanding high-rate capability of 75.3 mAh g-1 at 6000 mA g-1. Moreover, it shows that the NVP@C cathode is able to display a volumetric energy density of 54 Wh L-1 at 6000 mA g-1 (31 Wh L-1 for NVP bulk), as well as excellent cycling performance of 65.4 mAh g-1 after 1000 cycles at 2000 mA g-1. Furthermore, the NVP@C exhibits remarkable reversible capabilities of 81.9 mAh g-1 at a current density of 100 mA g-1 and 60.2 mAh g-1 at 1000 mA g-1 even at a low temperature of -15 °C. The structure of porous carbon frameworks combined with single crystal materials by in-situ polycondensation offers general guidelines for the design of sodium, lithium and potassium energy storage materials.

Identification and Potential Value of Candidate Genes in Patients With Non-obstructive Azoospermia.

OBJECTIVES: To explore the candidate genes involved in the pathogenesis of non-obstructive azoospermia (NOA) using bioinformatics analysis and experimental verification. METHODS: The gene expression profiles (GSE9210) were downloaded from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were identified. We performed function enrichment analyses, constructed protein-protein interaction (PPI) network and identified hub genes. Further, the miRNA-hub genes regulatory network was constructed. Finally, the expression level of CEP55 was verified using RT-qPCR and Western blot, and its diagnostic value was analyzed by the receiver operating characteristic (ROC) curve. RESULTS: 626 DEGs were identified, including 11 upregulated and 615 downregulated genes. Function enrichment analyses showed that these DEGs were significantly enriched in spermatogenesis, fertilization, meiotic cell cycle, flagellated sperm motility, sperm capacitation, spermatid nucleus differentiation and male meiotic nuclear differentiation. The top 10 hub genes were identified including CCNB2, BUB1, TOP2A, BIRC5, CENPF, PBK, NCAPG, DLGAP5, NUF2 and CEP55. In the miRNAs prediction, the hsa-miRNA-449a, hsa-miRNA-34c-5p and hsa-miRNA-34b-5p may be implicated in NOA. In the validation stage, the expression level of CEP55 was significantly decreased in patients with NOA compared to patients with OA. ROC analysis showed that CEP55 had a good diagnostic value for NOA and the combination of CEP55, FSH and mean testicular volume enhanced the prediction performance. CONCLUSIONS: This study identified key genes associated with NOA and their biological functions. Furthermore, CEP55 might play an important role in the pathogenesis of NOA, which will provide novel insights into the targeting therapy of NOA.

Corrigendum: Genomic Investigation Reveals a Community Typhoid Outbreak Caused by Contaminated Drinking Water in China, 2016.

[This corrects the article DOI: 10.3389/fmed.2022.753085.].

AroC, a chorismate synthase, is required for the formation of Edwardsiella tarda biofilms.

Biofilms contribute to the resistance of Edwardsiella tarda to antibiotics and host immunity. AroC in the shikimate pathway produces chorismate to synthesize crucial intermediates such as indole. In this study, the differences between biofilms produced by aroC mutants (△aroC), wild-type (WT) strains, and △aroC complementary strains (C△aroC) were detected both in vitro with 96-well plates, tubes, or coverslips and in vivo using a mouse model of subcutaneous implants. When examining potential mechanisms, we found that the diameters of the movement rings in soft agar plates and the flagellar sizes and numbers determined by silver staining were all lower for △aroC than for WT and C△aroC. Moreover, qRT-PCR showed that the transcription levels of flagellar synthesis genes, fliA and fliC, were reduced in △aroC. AroC, FliC, or FliA may accompany the motility of △aroC strains. In addition, compared with the WT and C△aroC, the amounts of indole in △aroC were significantly decreased. Notably, the formation of biofilms by these strains could be promoted by exogenous indole. Therefore, the aroC gene could affect the biofilm formation of E. tarda concerning its impact on flagella and indole.

Genomic Investigation Reveals a Community Typhoid Outbreak Caused by Contaminated Drinking Water in China, 2016.

Typhoid fever is a life-threatening disease caused by Salmonella enterica serovar Typhi (S. Typhi) and remains a significant public health burden in developing countries. In China, typhoid fever is endemic with a limited number of reported outbreaks. Recently, Chinese local Center for Disease Prevention and Control is starting to apply whole genome sequencing for tracking the source of outbreak isolates. In this study, we conducted a retrospective investigation into a community outbreak of typhoid fever in Lanling, China, in 2016. A total of 26 S. Typhi isolates were recovered from the drinking water (n = 1) and patients' blood (n = 24) and stool (n = 1). Phylogenetic analysis indicated the persistence of the outbreak isolates in drinking water for more than 3 months. The genomic comparison demonstrated a high similarity between the isolate from water and isolates from patients in their genomic content, virulence gene profiles, and antimicrobial resistance gene profile, indicating the S. Typhi isolate from drinking water was responsible for the examined outbreak. The result of pulsed-field gel electrophoresis (PFGE) revealed these isolates had identical PFGE pattern, indicating they are clonal variants. Additionally, phylogeographical analysis of global S. Typhi isolates suggested the outbreak isolates are evolutionarily linked to the isolates from the United Kingdom and Vietnam. Taken together, this study highlights the drinking water and international travel as critical control points of mitigating the outbreak, emphasizing the necessity of regular monitoring of this pathogen in China.

Long non­coding RNA CCAT1 enhances human non­small cell lung cancer growth through downregulation of microRNA­218.

Long non­coding RNAs (lncRNAs) have critical functions in non­small cell lung cancer (NSCLC) growth. In the present study, we showed that lncRNA­CCAT1 was upregulated in NSCLC tissues. High expression of lncRNA­CCAT1 was related to tumor growth and reduced survival rate. We used short hairpin RNAs (shRNAs) to inhibit the expression of lncRNA­CCAT1 in NSCLC cells. In vitro and in vivo results demonstrated that lncRNA­CCAT1 knockdown suppressed tumor proliferation and induced apoptosis. Furthermore, microRNA­218 (miR­218) was confirmed as an effective target of lncRNA­CCAT1 in NSCLC. B lymphoma Mo­MLV insertion region 1 homolog (BMI­1), which served as a downstream target of miR­218, was also inhibited by lncRNA­CCAT1 knockdown. In conclusion, the present study indicated that upregulation of lncRNA­CCAT1 in NSCLC is associated with tumor malignant potential. lncRNA­CCAT1 enhances tumor growth in NSCLC by directly inhibiting miR­218 and indirectly increasing BMI­1 expression.

Target genes directly regulated by Eha are required for Edwardsiella tarda survival within macrophages.

Eha is a virulence regulator in Edwardsiella tarda (E. tarda). The present study examined how Eha regulated its target genes to affect the bacterial survival within the cells. We constructed the reporter a pGEX-4T-ehaflag plasmid expressing Eha tagged at its C terminus with the flag epitope, and introduced the plasmid into an eha mutant ET13 strain, and obtained a Cehaflag strain. The expression and activity of an EhaFlag fusion protein restored the survival of the Cehaflag as the wild type in macrophages by Western blotting and intracellular survival experiments. We used a monoclonal anti-Flag antibody to precipitate EhaFlag-DNA complexes using chromatic immunoprecipitation (ChIP). We then designed primers based on the differentially-expressed genes identified from RNA-sequencing, and identified ten Eha-interacting genes by qPCR. We amplified the promoter regions of the ten genes and the eha gene from ET13 strain by PCR, constructed pBD-PtargetlacZ and pBD-PehalacZ plasmids. The eha gene directly and positively regulated these target genes, and be negatively auto-regulated by Eha in E. tarda, as determined by comparing their ß-Galactosidase activities. These target genes were distributed in the categories involved in the bacterial growth, movement and resistance to H2O2 or acid. We further constructed a ETATCC_RS15225 mutant (△dcuA1), a ETATCC_ RS14855 mutant (△flgK) anda ETATCC_RS07650 mutant (ΔtnaA), and a partial complementary strains of △eha-tnaA and △eha-flgK and the complementary strains of CΔflgK and CΔtnaA. The ETATCC_RS15225 gene probably encoded a transporter protein DcuA1 at outer membrane with SDS-PAGE and RT-PCR. The ETATCC _RS14855 gene probably encoded FlgK protein and affected the bacterial motility. The ETATCC_RS07650 gene encoded Tryptophanase, which affected the bacterial survival within macrophages. With the assistance of these above strains, our results showed that the eha gene was able to regulate the ETATCC_RS15225 gene to express its outer membrane protein DcuA1, the ETATCC _RS14855 gene to control the flagellar motility and the ETATCC_RS07650 to affect the bacterial survival within macrophages. With the combination of other functions of above three genes, our results suggested that Eha directly regulates the target genes to affect E. tarda to survive within the cells.

Guajadial inhibits NSCLC growth and migration following activation of the VEGF receptor-2.

Guajadial, one of natural dialdehyde meroterpenoids, demonstrated significant antineoplasmic activity. The present research was to investigate the inhibitory effects of guajadial by using two NSCLC cells (A549 and H1650) proliferation and migration. Western blotting was employed to explore the underlying mechanisms of VEGF receptor (VEGFR)2-mediated. This research indicated that guajadial not only inhibited endothelial cell proliferation and migration but also suppress tumor growth in human NSCLC xenograft mouse models. It is also suggested that guajadial inhibited A549 proliferation via blocking the Ras/MAPK pathway.

miR-149 suppresses human non-small cell lung cancer growth and metastasis by inhibiting the FOXM1/cyclin D1/MMP2 axis.

Valid evidence has demonstrated that microRNAs have critical functions in cancer genesis and tumor progression. In the present study, aberrant expression of microRNA-149 (miR-149) was confirmed in non-small cell lung cancer (NSCLC) tissues. Low expression of miR-149 was associated with malignant clinical features and poor survival. Gain- and loss-of-function experiments demonstrated that miR-149 inhibited NSCLC tumor growth and metastasis in vitro and in vivo. Furthermore, oncogenic transcription factor FOXM1 was confirmed as a direct target of miR-149 in NSCLC. Cyclin D1 and MMP2 served as downstream targets of FOXM1 and were also inhibited by miR-149. In summary, the present study indicated that downregulation of miR-149 in NSCLC predicted poor clinical outcomes. miR-149 suppresses tumor growth and metastasis in NSCLC by inhibiting the FOXM1/cyclin D1/MMP2 signaling pathway.

Overexpression of TSPAN8 Promotes Tumor Cell Viability and Proliferation in Nonsmall Cell Lung Cancer.

BACKGROUND: Overexpression of TSPAN8 has been involved in several epithelial cancers and TSPAN8 can form a complex with a variety of proteins to participate in several import cellular functions. However, the effects of TSPAN8 in nonsmall cell lung cancer (NSCLC) remain unclear. MATERIALS AND METHODS: In this study, the authors determined the expression of TSPAN in several NSCLC cell lines (95C, A549, H1299, and 95D) and human bronchial epithelial (HBE) cells. Furthermore, the authors investigated the biological function of TSPAN8 in NSCLC cell lines using gain-of-function and loss-of-function assays, as well as the underlying mechanisms. RESULTS: TSPAN8 was found to be overexpressed in NSCLC cells compared with normal HBE cells, of which the expression in H1299 is the highest and, in 95C, it is relatively lowest. Functional assays indicated that knockdown of TSPAN8 in H1299 remarkably reduced cell viability and proliferation, while overexpression of TSPAN8 in 95C dramatically enhanced cell viability and proliferation. In addition, TSPAN8 knockdown led to G1 phase arrest and apoptosis by downregulating CDK2, CDK4, and Cyclin D1 and upregulating Bax and PARP. CONCLUSIONS: These results provide evidence that TSPAN8 may contribute to the pathogenesis of lung cancer by promoting cell viability and proliferation. TSPAN8 silencing may provide a potential therapeutic intervention for the treatment of NSCLC.

Explore the reaction mechanism of the Maillard reaction: a density functional theory study.

The mechanism of Maillard reaction has been investigated by means of density functional theory calculations in the gaseous phase and aqueous solution. The Maillard reaction is a cascade of consecutive and parallel reaction. In the present model system study, glucose and glycine were taken as the initial reactants. On the basis of previous experimental results, the mechanisms of Maillard reaction have been proposed, and the possibility for the formation of different compounds have been evaluated through calculating the relative energy changes for different steps of reaction under different pH conditions. Our calculations reveal that the TS3 in Amadori rearrangement reaction is the rate-determining step of Maillard reaction with the activation barriers of about 66.7 and 68.8 kcal mol(-1) in the gaseous phase and aqueous solution, respectively. The calculation results are in good agreement with previous studies and could provide insights into the reaction mechanism of Maillard reaction, since experimental evaluation of the role of intermediates in the Maillard reaction is quite complicated.