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Bisphenol A (BPA) is widely exposed in populations worldwide and has negative effects on spermatogenesis both in animals and humans. The homeostasis of the actin cytoskeleton in the spermatogenic epithelium is crucial for spermatogenesis. Actin cytoskeleton destruction in the seminiferous epithelium is one of the important reasons for BPA‐induced spermatogenesis disorder. However, the underlying molecular mechanisms remain largely unexplored. Herein, we explored the role and mechanism of Rsad2, an interferon‐stimulated gene in BPA‐induced actin cytoskeleton disorder in mouse GC‐2 spermatocyte cell lines. After BPA exposure, the actin cytoskeleton was dramatically disrupted and the cell morphology was markedly altered accompanied by a significant increase in Rsad2 expression both in mRNA and protein levels in GC‐2 cells. Furthermore, the phalloidin intensities and cell morphology were restored obviously when interfering with the expression of Rsad2 in BPA‐treated GC‐2 cells. In addition, we observed a significant decrease in intracellular ATP levels after BPA treatment, while the ATP level was obviously upregulated when knocking down the expression of Rsad2 in BPA‐treated cells compared to cells treated with BPA alone. Moreover, Rsad2 relocated to mitochondria after BPA exposure in GC‐2 cells. BPA promoted Rsad2 expression by activating type I IFN‐signaling in GC‐2 cells. In summary, Rsad2 mediated BPA‐induced actin cytoskeletal disruption in GC‐2 cells, which provided data to reveal the mechanism of BPA‐induced male reproductive toxicity. BPA induces actin cytoskeleton disruption in the seminiferous epithelium. However, the underlying molecular mechanisms remain unclear. Herein, we found that Rsad2 was required for BPA‐induced cytoskeletal abnormality in GC‐2 cells. Mechanistically, BPA promoted Rsad2 expression by activating type I IFN‐signaling. Afterward, Rsad2 relocated to mitochondria, leading to a decrease in intracellular ATP levels and subsequently resulting in actin cytoskeletal disruption. Taken together, Rsad2 mediated BPA‐induced actin cytoskeleton disorder in GC‐2 cells. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV) infection, has been a serious threat to the pork industry worldwide and continues to bring significant economic loss. Current vaccination strategies offer limited protection against PRRSV transmission, highlighting the urgent need for novel antiviral approaches. In the present study, we reported for the first time that betulonic acid (BA), a widely available pentacyclic triterpenoids throughout the plant kingdom, exhibited potent inhibition on PRRSV infections in both Marc-145 cells and primary porcine alveolar macrophages (PAMs), with IC50 values ranging from 3.3 µM to 3.7 µM against three different type-2 PRRSV strains. Mechanistically, we showed that PRRSV replication relies on energy supply from cellular ATP production, and BA inhibits PRRSV infection by reducing cellular ATP production. Our findings indicate that controlling host ATP production could be a potential strategy to combat PRRSV infections, and that BA might be a promising therapeutic agent against PRRSV epidemics. [ABSTRACT FROM AUTHOR]

The discovery of a lead compound against Candida albicans is urgently needed because of the lack of clinically available antifungal drugs and the increase in drug resistance. Herein, a β-carboline alkaloid methylaervine (MET) exhibited potential activity against C. albicans (MIC = 16–128 μg/mL), no hemolytic toxicity, and a low tendency to induce drug resistance. An antifungal mechanism study indicated that MET effectively inhibited the biofilm formation and disrupted the mature biofilm. Moreover, filamentation formation and spore germination were also weakened. The electron microscopy analysis revealed that MET could damage the cell structure, including the cell wall, membrane, and cytoplasm. In particular, the permeability and integrity of the cell membrane were destroyed. When it entered the fungi cell, it interfered with the redox homeostasis and DNA function. Overall, MET can inhibit the growth of C. albicans from multiple channels, such as biofilm, filamentation, cell structure, and intracellular targets, which are difficult to mutate at the same time to generate drug resistance. This work provides a promising lead compound for the creation of new antifungal agents against C. albicans. [ABSTRACT FROM AUTHOR]

In response to the complex working conditions and poor driving stability of high-clearance self-propelled sprayers, a nonlinear model of the chassis power system was established based on the independently controllable torque of each wheel of the developed electric sprayer. A layered-architecture chassis drive control strategy was formulated, and a stability control framework comprising an instability judgment module, an upper controller, and a lower controller was constructed based on the analysis of the impact of the centroid slip angle, the yaw rate, and the wheel slip rate on driving stability. An ideal reference model was established based on the seven-degree-of-freedom model of the sprayer, and the current state of the sprayer body was determined using the instability judgment module. A drive anti-slip controller and a yaw moment controller based on fuzzy PID theory and sliding mode control theory were designed. Additionally, an optimal torque distribution algorithm was developed based on tire characteristics to rationally allocate drive torque to each wheel, ensuring the stability of the sprayer during operation. Simulation tests were conducted using MATLAB/Simulink to evaluate the sprayer under four different driving conditions during transport and field operations. The test results showed that the "SMC + optimal distribution" control method in the chassis stability control strategy reduced the maximum deviations of the yaw rate and centroid slip angle by an average of 89.5% and 13.6%, respectively, compared to no control. The wheel slip rate during straight driving was well maintained at around 15%, enhancing the driving stability of the sprayer. [ABSTRACT FROM AUTHOR]

Background: Plastic-based products are ubiquitous due to their tremendous utility in our daily lives. Nanoplastic (NP) and microplastic (MP) pollution has become a severe threat to the planet and is a growing concern. It has been widely reported that polystyrene (PS) MPs are severely toxic to the male reproduction system, with effects including decreased sperm parameters, impaired spermatogenesis, and damaged testicular structures. However, the molecular mechanisms for impaired spermatogenesis remain poorly understood. Methods: C57BL/6 male mice were treated with PS-NPs (80 nm) and PS-MPs (5 μm) by oral gavage every day for 60 days. A series of morphological analyses were completed to explore the influence of PS-NP and PS-MP exposure on the testes. Compared to other cell types in the seminiferous tubule, PS-NP and PS-MP exposure can lead to decreased spermatocytes. Then, more refined molecular typing was further performed based on gene expression profiles to better understand the common and specific molecular characteristics after exposure to PS-NPs and PS-MPs. Results: There were 1794 common DEGs across the PS-NP groups at three different doses and 1433 common DEGs across the PS-MP groups at three different doses. GO and KEGG analyses of the common DEGs in the PS-NP and PS-MP groups were performed to enrich the common and specific functional progress and signaling pathways, including 349 co-enriched GO entries and 13 co-enriched pathways. Moreover, 348 GO entries and 33 pathways were specifically enriched in the PS-NP group, while 526 GO entries and 15 pathways were specifically enriched in the PS-MPs group. Conclusions: PS-NPs were predominantly involved in regulating retinoic acid metabolism, whereas PS-MPs primarily influenced pyruvate metabolism and thyroid hormone metabolism. Our results highlight the different molecular mechanisms of PS-NPs and PS-MPs in the impairment of spermatogenesis in male mammals for the first time, providing valuable insights into the precise mechanisms of PS-NPs and PS-MPs in male reproduction. [ABSTRACT FROM AUTHOR]

Objective To investigate the changes in morphological characteristics of sperm flagellum and in sperm motility parameters in Fscb/Cabyr double gene knockout (Fscb/Cabyr--/--) mice. Methods The testis and epididymis tissues of male mice with different genotypes were collected and the organ-to-body weight ratio was calculated. The changes of spermatogenesis function in the testicular tissues of the mice were observed using HE staining. The parameters of sperm motility were analyzed using a computer-aided sperm analysis (CASA) system, and sperm capacitation and acrosome reaction were observed with in vitro culture. The morphological characteristics of the sperms under optical microscope and electron microscope were compared among the mice with different genotypes. Results No significant difference was found in sperm density in the epididymis among the mice with 5 different genotypes. The progressive rate (PR) and motility rate (PR+NP), mean curvilinear velocity (VCL), mean straight-line velocity (VSL), mean path velocity (VAP), beat cross frequency (BCF) and amplitude of lateral head displacement (ALH) of the sperms were significantly lower in Fscb/Cabyr--/-- and Fscb/Cabyr++/-- mice than in the wild-type mice (P < 0.01). The Fscb/Cabyr--/-- and Fscb/Cabyr++/-- mice showed significantly lowered percentages of normal sperms 1.45)% and (53.22±3.51)%] as compared with the wild-type mice and significantly higher rates of sperm tail folding] than the wild-type mice. Ultrastructural observation of the sperm flagellum in Fscb/Cabyr--/-- and Fscb/Cabyr++/-- mice revealed that the longitudinal columns of the fibrous sheath disappeared in most of the sperms with disorganized circumferential ribs and heterogeneous reduction of the electron density in the sperms; some sperms showed disruption of the axoneme structure in the flagella with occasional disappearance and dissociation of the doublet microtubules at the center. Conclusion Both Cabyr single gene knockout and Fscb/Cabyr double gene knockout can cause significant abnormalities in flagellum morphology and motility of the sperms in male mice. CABYR protein is essential for maintaining normal fibrous sheath structure and normal motility of the sperm flagella.

Objective To construct a mouse model of targeted knockout of Fscb/Cabyr dual genes by using the CRISPR/Cas9 system, in order to found a basis for the study of the role of FSCB and CABYR protein complexe in sperm flagellum movement and capacitation. Methods The targeting sites for CRISPR/Cas9 system were designed according to the full-length sequence of Cabyr gene. Two pairs of single-guide RNA (sgRNA) were chemically synthesized, and then inserted into the linearized plasmid pUC57-T7-gRNA to express sgRNA. The Cas9 RNA and sgRNA were transcribed in vitro by T7 RNA polymerase, and the obtained products were mixed in proper proportion and microinjected into the fertilized eggs of C57BL/6J mice to obtain the primary mouse (F0) with Cabyr gene knockout. After the identification of gene mutation by PCR, 2 chimeric females mice and background mice were selected for backcross breeding, and the Cabyr gene knockout F1 heterozygous mice were obtained. Then they were cross bred with the previously established Fscb heterozygous mice, and finally the Fscb/Cabyr (--/--) double gene knockout homozygous mice were obtained. The deletions of Fscb/Cabyr proteins in double gene knockout homozygous mice were detected by Western blotting and immunofluorescence assay. Results Seven positive F1 mice (3 males and 4 females) with deletion of the Cabyr gene fragment and frame shifting were obtained. The homozygous Cabyr knockout mice were obtained by mating the heterozygous male mice with the female mice. Fscb/Cabyr (--/--) double gene knockout homozygous mice were obtained by repeated cross breeding of Fscb and Cabyr heterozygous mice. Western blot analysis indicated that the expression of FSCB and CABYR protein disappeared in the testis and spermatozoa of the male mice. Immunofluorescence assay showed that there was no fluorescence expression of FSCB and CABYR protein in the principle piece of the sperm flagella. Conclusion Fscb/Cabyr (--/--) double gene knockout homozygous mice are obtained by using the CRISPR/Cas9 system and cross breeding, which laid the foundation of animal model for further study on the function of FSCB and CABYR protein complex.

Objective To explore the effect of ten-eleven translocation family 1 (TET1) gene expression on the migration and invasion of lung cancer cells and investigate its underlying mechanism. Methods The expression level of TET1 gene in normal human bronchial epithelial (HBE) cells and lung cancer cell lines (H460, H358, 95D and LTEP-a-2) was detected by quantitative RT-PCR. The effects of different expression level (overexpression and knockdown) of TET1 gene on the migration and invasion ability in lung cancer cells was analyzed by wound healing experiment and Transwell assay. The mRNA and protein expression levels of downstream regulatory gene β-catenin (CTNNB1) and MMP7 were detected by quantitative RT-PCR and Western blotting to determine the possible signal pathway. Results The expression level of TET1 gene was decreased in lung cancer cells compared with normal HBE cells (P < 0.01). Overexpression of TET1 gene significantly inhibited the migration and invasion of lung cancer cells, while its knockdown enhanced the invasion and migration abilities of lung cancer cells (P < 0.01). The mRNA and protein expression levels of CTNNB1 and MMP7 were decreased after TET1 overexpression, while the mRNA and protein levels of the 2 molecules were increased after knockdown (P < 0.01). Blockade of CTNNB1 by its inhibitor significantly diminished the effect of TET1 on the migration and invasion of lung cancer cell lines. Conclusion The expression level of TET1 is down-regulated in lung cancer cell lines. TET1 gene may inhibit the migration and invasion of lung cancer cells mainly through Wnt/β-catenin signaling pathway and its key downstream genes CTNNB1 and MMP7.

Objective To investigate the mRNA expression of SMAD9 in lung adenocarcinoma, and its relationship with clinicopathological parameters, survival and prognosis, so as to provide some valuable references for the screening and postoperative survival. Methods The expression of SMAD9 in tumor tissues (510 cases) and adjacent normal tissues (59 cases) of lung adenocarcinoma patients and its relationship with clinicopathological parameters and postoperative survival were analyzed based on the Cancer Genome Atlas (TCGA) database. The expression of SMAD9 in tumor tissues and adjacent normal tissues of lung adenocarcinoma patients, HBE malignant transformed cells induced by 3-MCA and lung adenocarcinoma cells were detected by quantitative real-time RT-PCR (qRT-PCR) and Western blotting. The effect of SMAD9 overexpression and interference on cell proliferation was detected by CCK-8 assay. Results The mRNA expression analysis of SMAD9 in TCGA database showed that the expression of SMAD9 was significantly lower in lung adenocarcinoma than adjacent normal tissues (P < 0.01). The results of qRT-PCR and Western blotting also showed that the expression of SMAD9 was significantly down-regulated in 3-MCA-induced HBE malignant transformed cells and lung adenocarcinoma tissues and cell lines (P < 0.05). The down-regulation of SMAD9 expression was closely correlated with N grade and tumor grade (P < 0.05). ROC curve analysis indicated that the mRNA expression of SMAD9 was a sensitive indicator of lung adenocarcinoma (AUC=0.903, 95%CI=0.868~0.938, P < 0.001). Kaplan-Meier analysis displayed that the survival time of patients with low expression of SMAD9 was significantly shorter than those with high expression (P < 0.05). Overexpression of SMAD9 inhibited the proliferation of lung adenocarcinoma cells (P < 0.01), and the proliferation of HBE cells was significantly enhanced after interfering with SMAD9 expression as compared with the control cells (P < 0.01). Conclusion SMAD9 gene expression is down-regulated in the incidence and development of lung adenocarcinoma. The expression of SMAD9 may be of great value for the screening and postoperative survival of lung adenocarcinoma patients.

Bisphenol A (BPA), a typical endocrine disruptor, is known to have various adverse effects on the male reproductive system. However, the toxic effects and mechanisms of low-dose BPA have not yet been fully explored. In this study, male Kunming mice were orally administered low-dose BPA (0.03, 0.3 and 3 mg/kg/d) for ten consecutive weeks. Pathological sections of testicular tissue showed no significant morphological differences after BPA exposure. An analysis of the functional parameters of sperm revealed that exposure to low-dose BPA significantly decreased sperm motility, chemotaxis, and the acrosome reaction. An in vitro BPA exposure model combined with an omics data analysis showed that the olfactory receptor-related pathway was significantly enriched after BPA treatment. Subsequent experiments verified the reduced mRNA level of a novel olfactory receptor gene, Olfr25, in vivo and in vitro exposure models. Meanwhile, exposure to low-dose BPA reduced the intracellular calcium ion concentration and the mRNA levels of pore-forming subunits of the CatSper channel in sperm. Importantly, the knockdown of Olfr25 inhibited calcium ion levels and CatSper subunit expression in GC-2 cells. Olfr25 overexpression attenuated the BPA-induced downregulation of CatSper subunit expression in GC-2 cells. These findings indicate that Olfr25 might participate in low-dose BPA-induced sperm dysfunction by affecting the CatSper-Ca2+ signaling pathway. This study reveals a new mechanism underlying the effects of low-dose BPA on sperm function and provides a reference for assessing the safety of low-dose BPA exposure. [ABSTRACT FROM AUTHOR]

SUMMARY: The petals of rose (Rosa sp.) flowers determine the ornamental and industrial worth of this species. The number of petals in roses was previously shown to be subject to fluctuations in ambient temperature. However, the mechanisms by which rose detects and responds to temperature changes are not entirely understood. In this study, we identified short interstitial telomere motifs (telo boxes) in the second intron of AGAMOUS (RcAG) from China rose (Rosa chinensis) that play an essential role in precise temperature perception. The second intron of RcAG harbors two telo boxes that recruit telomere repeat binding factors (RcTRBs), which interact with CURLY LEAF (RcCLF) to compose a repressor complex. We show that this complex suppresses RcAG expression when plants are subjected to low temperatures via depositing H3K27me3 marks (trimethylation of lysine 27 on histone H3) over the RcAG gene body. This regulatory mechanism explains the low‐temperature‐dependent decrease in RcAG transcript levels, leading to the production of more petals under these conditions. Our results underscore an interesting intron‐mediated regulatory mechanism governing RcAG expression, enabling rose plants to perceive temperature cues and establish petal numbers. Significance Statement: Our research underscores a new intron‐mediated regulatory mechanism governing RcAG expression, enabling rose plants to perceive temperature cues and establish petal numbers. It might be of great interest to a broad range of readers including environmental response, epigenetic modification, floral organ development, and other researchers in related fields. [ABSTRACT FROM AUTHOR]

The structural, elastic, piezoelectric, and electronic properties of Li-doped K0.5Na0.5NbO3 (K0.5−xNa0.5−yLix+yNbO3, KNN-L) are calculated. The properties of KNN-L are related to the Li-doping content and the replaced K or Na atoms. The bulk modulus, the shear modulus, and Young's modulus of KNN-L are mostly higher than those of KNN, and the hardness value increases. The Poisson ratio of KNN-L is lower than that of most KNN, and the ductility is reduced. All doped structures are direct band gap semiconductors. K0.5Na0.375Li0.125NbO3 has the largest piezoelectric charge constant, d33 = 44.72 pC/N, in the respective structures, which is 1.5 fold that of K0.5Na0.5NbO3 (29.15 pC/N). The excellent piezoelectric performance of Li-doping KNN-L was analyzed from the insights of elastic and electronic properties. [ABSTRACT FROM AUTHOR]

In the digital transformation era, digitalization integrates deeply into production, bolstering output efficiency and economic value. Through stochastic frontier analysis (SFA), this research positions digitalization as an input in the production function, dissecting its elasticity impact on capital, labor, and output. The effect of digitalization on total factor productivity change (TFPC) is explained by comparing TFPC with and without digitalization. Findings reveal that digitalization's integration into economic growth displays a U-shaped trajectory, with initial productivity setbacks transitioning to long-term benefits as industries adapt. The periodic complementarity and substitution between digitalization and labor, along with a weak substitution relationship with capital, illustrate that, as a production factor, digitalization dynamically interacts with other factors, both complementing and substituting them. This dynamic interplay highlights the intricate role that digitalization plays within the production function. Furthermore, digitalization has played a crucial role in China's TFP growth, which also highlights the lack of other technological progress. Meanwhile, the pace of digital transformation presents scalability challenges, evident in the fluctuating scale efficiency change (SEC). Policymakers are advised to address these early stage challenges through supportive measures, ensuring smoother digital transitions. Concurrently, industries should embrace this non-linear transformation, emphasizing adaptability to maximize digitalization's long-term advantages. [ABSTRACT FROM AUTHOR]