Establishment of a Secretory Protein-Inducible CRISPR/Cas9 System for Nosema bombycis in Insect Cells.

Gene editing techniques are widely and effectively used for the control of pathogens, but it is difficult to directly edit the genes of Microsporidia due to its unique spore wall structure. Innovative technologies and methods are urgently needed to break through this limitation of microsporidia therapies. Here, we establish a microsporidia-inducible gene editing system through core components of microsporidia secreted proteins, which could edit target genes after infection with microsporidia. We identified that Nosema bombycis NB29 is a secretory protein and found to interact with itself. The NB29-N3, which lacked the nuclear localization signal, was localized in the cytoplasm, and could be tracked into the nucleus after interacting with NB29-B. Furthermore, the gene editing system was constructed with the Cas9 protein expressed in fusion with the NB29-N3. The system could edit the exogenous gene EGFP and the endogenous gene BmRpn3 after overexpression of NB29 or infection with N. bombycis.

Ilicicolin C suppresses the progression of prostate cancer by inhibiting PI3K/AKT/mTOR pathway.

Aberrant activation of the PI3K/AKT pathway is a driving factor in the development of prostate cancer. Therefore, inhibiting the function of the PI3K/AKT signaling pathway is a strategy for the treatment of prostate cancer. Ilicicolin C is an ascochlorin derivative isolated from the coral-derived fungus Acremonium sclerotigenum GXIMD 02501. Which has anti-inflammatory activity, but its activity against prostate cancer has not yet been elucidated. MTT assay, plate clone-formation assay, flow cytometry and real-time cell analysis technology were used to detect the effects of ilicicolin C on cell viability, proliferation, apoptosis and migration of prostate cancer cells. Molecular docking software and surface plasmon resonance technology were used to analyze the interaction between ilicicolin C and PI3K/AKT proteins. Western blot assay was performed to examine the changes in protein expression. Finally, QikProp software was used to simulate the process of ilicicolin C in vivo, and a zebrafish xenograft model was used to further verify the anti-prostate cancer activity of ilicicolin C in vivo. Ilicicolin C showed cytotoxic effects on prostate cancer cells, with the most significant effect on PC-3 cells. Ilicicolin C inhibited proliferation and migration of PC-3 cells. It could also block the cell cycle and induce apoptosis in PC-3 cells. In addition, ilicicolin C could bind to PI3K/AKT proteins. Furthermore, ilicicolin C inhibited the expression of PI3K, AKT and mTOR proteins and could also regulate the expression of downstream proteins in the PI3K/AKT/mTOR signaling pathway. Moreover, the calculations speculated that ilicicolin C was well absorbed orally, and the zebrafish xenograft model confirmed the in vivo anti-prostate cancer effect of ilicicolin C. Ilicicolin C emerges as a promising marine compound capable of inducing apoptosis of prostate cancer cells by counteracting the aberrant activation of PI3K/AKT/mTOR, suggesting that ilicicolin C may be a viable candidate for anti-prostate cancer drug development. These findings highlight the potential of ilicicolin C against prostate cancer and shed light on its mechanism of action.

The Impact of Skill Level on the Integration of Information and Post-Error Adjustment during Action Anticipation in Basketball.

The present study examined the impact of skill level on the integration of contextual prior information and kinematic information alongside post-error adjustment during action anticipation in basketball. Twenty-three collegiate basketball players and twenty-three control participants engaged in anticipating as quickly and accurately as possible the outcomes of free throws, utilizing video clips depicting basketball players' actions, both with and without contextual prior information. Anticipatory performance and the difference in anticipatory performance following errors and correct responses were analyzed based on skill level and the congruency of contextual prior information (none, congruent, and incongruent). The findings revealed that the congruency of contextual prior information significantly affects action anticipation, with skill level moderating this effect. Moreover, skill level influenced the congruency effect on accuracy discrepancies between post-error and post-correct trials during action anticipation, with controls showing greater sensitivity to previous trial performance compared to experts. These results provide further evidence for the notion that individuals employ Bayesian reliability-based strategies to integrate different information sources and underscore the role of skill level in adjusting anticipatory judgments following errors during action anticipation. These insights contribute to a deeper understanding of the cognitive and behavioral mechanisms that differentiate skill levels in action anticipation, potentially guiding the development of targeted training interventions.

Recent advances in flavonoid compounds for the treatment of prostate cancer.

Prostate cancer is a malignant epithelial tumor of the prostate gland and is the most common malignant tumor of the male genitourinary system. Pharmacological therapies, including chemotherapy and androgen deprivation therapy, play a key role in the treatment of prostate cancer. However, drug resistance and side effects limit the use of these drugs and so there is a need for new drug therapies for prostate cancer patients. Flavonoids, with their wide range of sources and diverse biological activities, have attracted much attention in the field of anti-tumor drug screening. In 2016, at least 58 flavonoids were reported to have anti-prostate cancer activity. In recent years, six additional flavonoid compounds have been found to have anti-prostate cancer potential. In this review, we have collected a large amount of evidence on the anti-prostate cancer effects of these six flavonoids, including a large number of cellular experiments and a small number of preclinical animal experiments. In addition, we predicted their drug-forming properties using Schrödinger's QikProp software and ADMETlab due to the lack of in vivo pharmacokinetic data for the six compounds. In conclusion, this review has fully confirmed the anti-prostate cancer effects of these six flavonoids, summarized their mechanisms of action and predicted their druggability. It provides a reference for the further development of these compounds into anti-prostate cancer drugs.

Midbrain node for context-specific vocalisation in fish.

Vocalizations communicate information indicative of behavioural state across divergent social contexts. Yet, how brain regions actively pattern the acoustic features of context-specific vocal signals remains largely unexplored. The midbrain periaqueductal gray (PAG) is a major site for initiating vocalization among mammals, including primates. We show that PAG neurons in a highly vocal fish species (Porichthys notatus) are activated in distinct patterns during agonistic versus courtship calling by males, with few co-activated during a non-vocal behaviour, foraging. Pharmacological manipulations within vocally active PAG, but not hindbrain, sites evoke vocal network output to sonic muscles matching the temporal features of courtship and agonistic calls, showing that a balance of inhibitory and excitatory dynamics is likely necessary for patterning different call types. Collectively, these findings support the hypothesis that vocal species of fish and mammals share functionally comparable PAG nodes that in some species can influence the acoustic structure of social context-specific vocal signals.

Analysis of regulating activities of 5'-epiequisetin on proliferation, apoptosis, and migration of prostate cancer cells in vitro and in vivo.

Advanced prostate cancer has a poor prognosis, and it is urgent to develop new effective drugs. 5'-Epiequisetin is a tetramic acid derivative which was isolated from a marine sponge-derived fungus Fusarium equiseti in our previous study. In this study, 5'-epiequisetin showed cytotoxicity against four prostate cancer cell lines, namely, LNCaP, 22Rv1, DU145, and PC-3 cells, with the lowest IC50 value of 4.43 ± 0.24 µM in PC-3 cells. Further studies showed that it could dramatically regulate the clonal colony formation, apoptosis, and migration of PC-3 cells. In addition, flow cytometry data showed that 5'-epiequisetin could block the cell cycle at the G1 phase. Proteome profiler array and Western blot revealed that 5'-epiequisetin could regulate the expression of proteins responsible for cell proliferation, apoptosis, and migration. 5'-Epiequisetin regulated the expression of PI3K, Akt, phosphorylated Akt, and proteins which control the cell cycle. Meanwhile, 5'-epiequisetin upregulated expression of DR5 and cleave-caspase 3, which play important roles in the process of apoptosis. Moreover, when DR5 was silenced by small interfering RNA, the proportion of apoptotic cells induced by 5'-epiequisetin remarkably declined. In addition, 5'-epiequisetin downregulated the expression of survivin which plays a key role in the process of survival and apoptosis. 5'-Epiequisetin also impacted beta-catenin and cadherins, which were associated with cell migration. In addition, 5'-Epiequisetin significantly inhibited the progression of prostate cancer in mice, accompanied by regulating the protein expression of DR5, caspase 8, survivin, and cadherins in vivo. Taken together, these findings indicated that 5'-epiequisetin showed an anti-prostate cancer effect by inducing apoptosis and inhibiting cell proliferation and migration both in vitro and in vivo, suggesting a promising lead compound for the pharmacotherapy of prostate cancer.

Acoustic and postural displays in a miniature and transparent teleost fish, Danionella dracula.

Acoustic behavior is widespread across vertebrates, including fishes. We report robust acoustic displays during aggressive interactions for a laboratory colony of Danionella dracula, a miniature and transparent species of teleost fish closely related to zebrafish (Danio rerio), which are hypothesized to be sonic based on the presence of a hypertrophied muscle associated with the male swim bladder. Males produce bursts of pulsatile sounds and a distinct postural display - extension of a hypertrophied lower jaw, a morphological trait not present in other Danionella species - during aggressive but not courtship interactions. Females show no evidence of sound production or jaw extension in such contexts. Novel pairs of size-matched or -mismatched males were combined in resident-intruder assays where sound production and jaw extension could be linked to individuals. In both dyad contexts, resident males produced significantly more sound pulses than intruders. During heightened sonic activity, the majority of the highest sound producers also showed increased jaw extension. Residents extended their jaw more than intruders in size-matched but not -mismatched contexts. Larger males in size-mismatched dyads produced more sounds and jaw extensions compared with their smaller counterparts, and sounds and jaw extensions increased with increasing absolute body size. These studies establish D. dracula as a sonic species that modulates putatively acoustic and postural displays during aggressive interactions based on residency and body size, providing a foundation for further investigating the role of multimodal displays in a new model clade for neurogenomic and neuroimaging studies of aggression, courtship and other social interactions.

Analysis of Silver Nanoparticles for the Treatment and Prevention of Nucleopolyhedrovirus Affecting Bombyx mori.

Bombyx mori nucleopolyhedrovirus (BmNPV) causes major economic losses in sericulture. A number of agents have been employed to treat viral diseases. Silver nanoparticles (AgNPs) have wide applications in biomedical fields due to their unique properties. The anti-BmNPV effect of AgNPs has been evaluated, however, there are insufficient studies concerning its toxicity to other organisms and the environment. We chemically synthesized biocompatible BSA-AgNPs with a diameter range of 2-4 nm and characterized their physical properties. The toxicity of AgNPs towards cells and larvae with different concentrations was examined; the results indicated a biofriendly effect on cells and larvae within specific concentration ranges. The SEM observation of the surface of BmNPV after treatment with AgNPs suggested that AgNPs could destroy the polyhedral structure, and the same result was obtained by Coomassie blue staining. Further assays confirmed the weakened virulence of AgNPs-treated BmNPV toward cells and larvae. AgNPs also could effectively inhibit the replication of BmNPV in infected cells and larvae. In summary, our research provides valuable data for the further development of AgNPs as an antiviral drug for sericulture.

Kaempferol suppresses androgen-dependent and androgen-independent prostate cancer by regulating Ki67 expression.

PURPOSE: Kaempferol is a natural flavonoid that has been reported to be active against many cancers, including prostate cancer, breast cancer and colon cancer. In our previous study, we found kaempferol could act as a selective androgen receptor modulator, thereby suppress development of benign prostatic hyperplasia. This finding inspired us to further explore the effect and the mechanism of action of kaempferol on prostate cancer. METHODS: Plate clone formation assay was performed to detect the effect of kaempferol on cell proliferation. Flow cytometry was used to detect the impact of kaempferol on cell apoptosis and cell cycle distribution. qPCR, immunofluorescence staining, and enzyme-linked immunosorbent assay were used to detect the expression of gene and protein of Ki67 which is a biomarker of cell proliferation. RESULTS: In the present study, we found kaempferol could dramatically suppress androgen-dependent and androgen-independent prostate cancer cells proliferation and induce their apoptosis. Furthermore, we found that kaempferol induced cell cycle to be arrested at G1 phase in 22Rv1 cells but at S and G2 phase in PC-3 cells. In addition, we detected the mRNA and protein of Ki67 which is corresponding to the cell proliferation and found that kaempferol could significantly inhibit Ki67 expression at mRNA level but increase its expression at protein levels in both androgen-dependent and androgen-independent prostate cancer cells. CONCLUSION: Taken together, kaempferol inhibited the proliferation of androgen-dependent and androgen-independent prostate cancer cells by regulating the expression of Ki67. These findings further shed light on the mechanism of action of kaempferol on anti-prostate cancer.

Nosema bombycis microRNA-like RNA 8 (Nb-milR8) Increases Fungal Pathogenicity by Modulating BmPEX16 Gene Expression in Its Host, Bombyx mori.

The fungus Nosema bombycis causes significant economic losses via parasitism of an economically important insect. MicroRNAs (miRNAs) play important roles in regulating host and parasite gene expression via mRNA degradation or by inhibiting protein translation. To investigate whether microRNA-like RNAs (milRNAs) regulate N. bombycis pathogenesis and to better understand the regulatory mechanisms underlying infection, we constructed small RNA libraries from N. bombycis hyphae during the schizont proliferation period. Eleven novel milRNAs were determined by RNA sequencing and stem-loop reverse transcriptase PCR (RT-PCR) assays. Moreover, a virulence-associated milRNA, Nb-milR8, was identified as critical for N. bombycis proliferation by binding and downregulating expression of its target gene, BmPEX16, in the host during infection. Silencing of Nb-milR8 or overexpression of the target BmPEX16 gene resulted in increased susceptibility of Bombyx mori to N. bombycis infection. Taken together, these results suggest that Nb-milR8 is an important virulence factor that acts as an effector to suppress host peroxidase metabolism, thereby facilitating N. bombycis proliferation. These results provide important novel insights into interactions between pathogenic fungi and their hosts. IMPORTANCE A thorough understanding of fungal pathogen adaptations is essential for treating fungal infections. Recent studies have suggested that the role of small RNAs expressed in fungal microsporidia genomes are important for elucidating the mechanisms of fungal infections. Here, we report 11 novel microRNA-like RNAs (milRNAs) from the fungal microsporidium Nosema bombycis and identified NB-milRNAs that adaptively regulate N. bombycis proliferation. In addition, we demonstrate that N. bombycis modulates small RNA (sRNA)-mediated infection by encoding an Nb-miR8 that downregulates the expression of the host peroxidase metabolism protein BmPEX16, which is essential for peroxisome membrane biogenesis and peroxisome assembly. These results significantly contribute to our understanding of the pathogenic mechanisms of fungi, and especially microsporidia, while providing important targets for genetical engineering-based treatment of microsporidia.

The Biomechanical Response of the Cornea in Orthokeratology.

Orthokeratology has been widely used to control myopia, but the mechanism is still unknown. To further investigate the underlying mechanism of corneal reshaping using orthokeratology lenses via the finite element method, numerical models with different corneal curvatures, corneal thicknesses, and myopia reduction degrees had been developed and validated to simulate the corneal response and quantify the changes in maximum stress in the central and peripheral corneal areas during orthokeratology. The influence of the factors on corneal response had been analyzed by using median quantile regression. A partial eta squared value in analysis of variance models was established to compare the effect size of these factors. The results showed central and peripheral corneal stress responses changed significantly with increased myopia reduction, corneal curvature, and corneal thickness. The target myopia reduction had the greatest effect on the central corneal stress value (partial eta square = 0.9382), followed by corneal curvature (partial eta square = 0.5650) and corneal thickness (partial eta square = 0.1975). The corneal curvature had the greatest effect on the peripheral corneal stress value (partial eta square = 0.5220), followed by myopia reduction (partial eta square = 0.2375) and corneal thickness (partial eta square = 0.1972). In summary, the biomechanical response of the cornea varies significantly with the change in corneal conditions and lens designs. Therefore, the orthokeratology lens design and the lens fitting process should be taken into consideration in clinical practice, especially for patients with high myopia and steep corneas.

Modeling the Keys to Team's Success in the Women's Chinese Basketball Association.

The technical characteristics of women's basketball may differ from men's basketball, and there is a need to identify the key performance indicators (KPIs) that contribute to the success of women's teams. The aim of the current study was to examine and quantify the relationships between technical performance indicators and match outcome in elite women's basketball using both linear and non-linear statistical methods, the effectiveness of the two methods was compared as well. A total of 136 matches (n = 272 teams' observations) in the regular season of Women's Chinese Basketball Association (WCBA; season 2020-2021) were analyzed using multiple linear regression (MLR) and quantile regression (QR). Results showed that two-point percentage, offensive rebounds, assists and turnovers had significant effects on the match outcome for both MLR and QR analysis. No significant relationships were observed between match outcome and three-point percentage, steals, and fouls. The results between MLR and QR analysis were different in free-throw percentage, defensive rebounds and blocks. Current results highlighted QR analysis is an advanced statistical model more powerful than the traditional linear method for the identification of KPIs. The identified KPIs may help coaches to develop more specific training interventions and match strategies during match play.

Performance of Silicone Rubber Composites Filled with Aluminum Nitride and Alumina Tri-Hydrate.

In this study, silicone rubber (SR) composites were prepared with various amounts of aluminum nitride (AlN) and alumina tri-hydrate (ATH), and vinyl tri-methoxysilane (VTMS) was also introduced to prepare SR/ATH/AlN-VTMS composites for comparison. Compared to the SR/ATH composites, the SR/ATH/AlN composites with higher AlN loading exhibited higher breakdown strength and thermal conductivity, which were further improved by the addition of VTMS. Such results were related to the enhanced rubber-filler interfacial interactions from VTMS coupling, as demonstrated by scanning electron microscopy (SEM) analysis and the curing behaviors of the SR composites. Moreover, by replacing ATH with VTMS-coupled AlN, the SR/ATH/AlN-VTMS composites also exhibited lower dielectric loss along with an increased dielectric constant, suggesting the promising application of VTMS-coupled AlN as a filler for the preparation of the SR composites as high-voltage insulators.

Morphometric and hemodynamic parameter dataset for coronary artery aneurysms caused by atherosclerosis.

In comparison with intracranial aneurysm, there are relatively few investigations of coronary artery aneurysms (CAA). Coronary atherosclerosis is the first cause of CAA; therefore, it is necessary to providing as many details of clinical CAA caused by atherosclerosis as possible. The aim of the data is to provide morphometric and hemodynamic parameters of CAAs caused by atherosclerosis, as well as the demographics of patients with CAAs. Various morphometric parameters were obtained from the reconstructed epicardial coronary arterial trees of 61 patients while multiple hemodynamic parameters were determined from their computed flow fields. The data classified the CAAs into 4 types. All subjects in each group are listed in this data article. This data set support the main findings presented in the research article (Fan et al., 2019).

Morphometry and hemodynamics of coronary artery aneurysms caused by atherosclerosis.

BACKGROUND AND AIMS: Atherosclerosis is the first etiology of coronary artery aneurysm (CAA). It is, however, challenging to inhibit the development of CAA. The aim of the study is to carry out morphometric and hemodynamic analyses in epicardial coronary arteries of patients with CAAs caused by atherosclerosis. METHODS: Various morphometric parameters were obtained from the reconstructed epicardial coronary arterial trees of 61 patients and multiple hemodynamic parameters were determined from the computed flow fields. A two-step analysis method was proposed to enhance the risk assessment of CAAs, i.e., coronary artery bifurcation is the major risk factor for CAA followed by high aneurysm shape index (L/W, where L and W refer to the aneurysm length and maximum diameter, respectively). RESULTS: Patients with CAA covering a bifurcation have the higher occurrence (71% and 55% for L/W ≥ 2 and L/W < 2, respectively) of myocardial ischemia relevant to abnormal hemodynamic parameters in comparison with those with CAA in one vessel (43% and 40% for L/W ≥ 2 and L/W < 2, respectively). Patients with CAA of L/W ≥ 2 covering a bifurcation show the worst hemodynamic environment. CONCLUSIONS: Morphometric and hemodynamic studies support the two-step analysis method, which provides a clinical rationale for the noninvasive assessment of CAAs.