Bile acids promote the caveolae-associated entry of swine acute diarrhea syndrome coronavirus in porcine intestinal enteroids.

Intestinal microbial metabolites have been increasingly recognized as important regulators of enteric viral infection. However, very little information is available about which specific microbiota-derived metabolites are crucial for swine enteric coronavirus (SECoV) infection in vivo. Using swine acute diarrhea syndrome (SADS)-CoV as a model, we were able to identify a greatly altered bile acid (BA) profile in the small intestine of infected piglets by untargeted metabolomic analysis. Using a newly established ex vivo model-the stem cell-derived porcine intestinal enteroid (PIE) culture-we demonstrated that certain BAs, cholic acid (CA) in particular, enhance SADS-CoV replication by acting on PIEs at the early phase of infection. We ruled out the possibility that CA exerts an augmenting effect on viral replication through classic farnesoid X receptor or Takeda G protein-coupled receptor 5 signaling, innate immune suppression or viral attachment. BA induced multiple cellular responses including rapid changes in caveolae-mediated endocytosis, endosomal acidification and dynamics of the endosomal/lysosomal system that are critical for SADS-CoV replication. Thus, our findings shed light on how SECoVs exploit microbiome-derived metabolite BAs to swiftly establish viral infection and accelerate replication within the intestinal microenvironment.

A One-Step RPA-CRISPR Assay Using crRNA Based on Suboptimal Protospacer Adjacent Motif for Vibrio vulnificus Detection.

Vibrio vulnificus is a hazardous foodborne pathogen responsible for approximately 95% of seafood-related deaths. This highlights the urgent requirement for specialized detection tools to be developed and used by food enterprises and food safety authorities. The DETECTR (DNA endonuclease targeted CRISPR trans reporter) system that combines CRISPR/Cas and recombinase polymerase amplification (RPA) has been utilized to develop a molecular detection assay for V. vulnificus. However, because the incompatibility between RPA and Cas12a cleavage has not been addressed, it is a two-step assay that lacks convenience and presents contamination risk. Here, we developed a one-step RPA-CRISPR assay for V. vulnificus using a special crRNA targeting a sequence with a suboptimal protospacer adjacent motif (PAM). The entire assay, conducted at 37°C, takes only 40-60 min, yields results visualized under blue light, and exhibits exceptional specificity and sensitivity (detecting 4 pathogen genome copies per reaction). This study offers a valuable tool for detecting V. vulnificus, aiding in foodborne infection prevention, and exemplifies one-step RPA-CRISPR assays managing Cas-cleavage activity through PAM adjustments.

Advances in preparation and application of antibacterial hydrogels.

Bacterial infections, especially those caused by drug-resistant bacteria, have seriously threatened human life and health. There is urgent to develop new antibacterial agents to reduce the problem of antibiotics. Biomedical materials with good antimicrobial properties have been widely used in antibacterial applications. Among them, hydrogels have become the focus of research in the field of biomedical materials due to their unique three-dimensional network structure, high hydrophilicity, and good biocompatibility. In this review, the latest research progresses about hydrogels in recent years were summarized, mainly including the preparation methods of hydrogels and their antibacterial applications. According to their different antibacterial mechanisms, several representative antibacterial hydrogels were introduced, such as antibiotics loaded hydrogels, antibiotic-free hydrogels including metal-based hydrogels, antibacterial peptide and antibacterial polymers, stimuli-responsive smart hydrogels, and light-mediated hydrogels. In addition, we also discussed the applications and challenges of antibacterial hydrogels in biomedicine, which are expected to provide new directions and ideas for the application of hydrogels in clinical antibacterial therapy.

A recombinase polymerase amplification and Pyrococcus furiosus Argonaute combined method for ultra-sensitive detection of white spot syndrome virus in shrimp.

White spot disease (WSD) in shrimp is an acute infectious disease caused by white spot syndrome virus (WSSV). WSD has seriously threatened the security of shrimp farming, causing huge economic losses worldwide. As there is currently no effective treatment for WSD, developing early detection technologies for WSSV is of great significance for the prevention. In this study, we have established a detection method for WSSV using a combination of recombinase polymerase amplification (RPA) and Pyrococcus furiosus Argonaute (PfAgo). We have achieved a detection sensitivity of single copy per reaction, which is more sensitive than the previously reported detection methods. Additionally, we have demonstrated high specificity. The entire detection process can be completed within 75 min without the need for precise thermal cyclers, making it suitable for on-site testing. The fluorescence signal generated by the reaction can be quantified using a portable fluorescence detector or observed with the naked eye under a blue light background. This study provides an ultrasensitive on-site detection method for WSSV in shrimp aquaculture and expands the application of PfAgo in the field of aquatic disease diagnosis.

Optimized DOX Drug Deliveries via Chitosan-Mediated Nanoparticles and Stimuli Responses in Cancer Chemotherapy: A Review.

Chitosan nanoparticles (NPs) serve as useful multidrug delivery carriers in cancer chemotherapy. Chitosan has considerable potential in drug delivery systems (DDSs) for targeting tumor cells. Doxorubicin (DOX) has limited application due to its resistance and lack of specificity. Chitosan NPs have been used for DOX delivery because of their biocompatibility, biodegradability, drug encapsulation efficiency, and target specificity. In this review, various types of chitosan derivatives are discussed in DDSs to enhance the effectiveness of cancer treatments. Modified chitosan-DOX NP drug deliveries with other compounds also increase the penetration and efficiency of DOX against tumor cells. We also highlight the endogenous stimuli (pH, redox, enzyme) and exogenous stimuli (light, magnetic, ultrasound), and their positive effect on DOX drug delivery via chitosan NPs. Our study sheds light on the importance of chitosan NPs for DOX drug delivery in cancer treatment and may inspire the development of more effective approaches for cancer chemotherapy.

Rapid and Sensitive Genotyping of SARS-CoV-2 Key Mutation L452R with an RPA-PfAgo Method.

In the two years of COVID-19 pandemic, the SARS-CoV-2 variants have caused waves of infections one after another, and the pandemic is not ending. The key mutations on the S protein enable the variants with enhanced viral infectivity, immune evasion, and/or antibody neutralization resistance, bringing difficulties to epidemic prevention and control. In support of precise epidemic control and precision medicine of the virus, a fast and simple genotyping method for the key mutations of SARS-CoV-2 variants needs to be developed. By utilizing the specific recognition and cleavage property of the nuclease Argonaute from Pyrococcus furiosus (PfAgo), we developed a recombinase polymerase amplification (RPA) and PfAgo combined method for a rapid and sensitive genotyping of SARS-CoV-2 key mutation L452R. With a delicate design of the strategy, careful screening of the RPA primers and PfAgo gDNA, and optimization of the reaction, the method achieves a high sensitivity of a single copy per reaction, which is validated with the pseudovirus. This is the highest sensitivity that can be achieved theoretically and the highest sensitivity as compared to the available SARS-CoV-2 genotyping assays. Using RPA, the procedure of the method is finished within 1.5 h and only needs a minimum laboratorial support, suggesting that the method can be easily applied locally or on-site. The RPA-PfAgo method established in this study provides a strong support to the precise epidemic control and precision medicine of SARS-CoV-2 variants and can be readily developed for the simultaneous genotyping of multiple SARS-CoV-2 mutations.

H2S inhibits atrial fibrillation-induced atrial fibrosis through miR-133a/CTGF axis.

AIM: Atrial fibrillation (AF) is a common clinical arrhythmia and can cause a variety of complications. To study the therapeutic effect of H2S in atrial fibrosis and explore the important role of miR-133a, in vitro experiments in human atrial fibroblasts (HAFs) were conducted. METHODS: The fibrosis in HAFs was induced by Ang II. The expression levels of miR-133a and CTGF in HAFs were examined by qRT-PCR. The proliferation and migration of HAFs were detected by CCK-8 and cell scratch assays. The protein expressions of CTGF, collagen I, collagen III and α-SMA were detected by western blotting. The dual-luciferase reporter gene was used to detect the interaction between miR-133a and CTGF. RESULTS: The proliferation and migration of HAFs stimulated by Ang II were enhanced, the expression of miR-133a was reduced, and the levels of CTGF and fibrosis markers (collagen I, collagen III and α-SMA) were increased. Furthermore, H2S reduced fibrosis, proliferation and migration of HAFs induced by Ang II. Accordingly, overexpression of miR-133a inhibited the proliferation and migration ability on Ang II-induced HAFs, and decreased the protein expressions of related fibrosis markers and CTGF. Meanwhile, miR-133a inhibitor could reverse the inhibition effect of H2S on proliferation and migration in HAFs by Ang II-induced. By targeting CTGF, miR-133a inhibited the expression of CTGF. CONCLUSION: H2S improved myocardial cell fibrosis by significantly increasing the expression of miR-133a, and CTGF might be a potential target for miR-133a to play an important role in myocardial fibrosis.

p22phox promotes Ang-II-induced vascular smooth muscle cell phenotypic switch by regulating KLF4 expression.

NADPH oxidase (Nox) is the main source of reactive oxygen species in vascular diseases, which have been implicated in promoting VSMCs phenotypic switch. P22phox, the indispensable component of the complex Nox, is required for their activity and stability. Krüppel-like factor 4 (KLF4) is an important transcriptional regulator of VSMCs phenotypic switch. Both KLF4 and p22phox are involved in the proliferation, migration and differentiation of VSMC. This study aims to determine whether and how p22phox regulates KLF4 expression in phenotypic switching of VSMCs. In cultured primary rat VSMCs, we noticed that the expression of P22phox was significantly increased in combination with VSMCs phenotypic switch and up-regulated KLF4 expression in Ang-II-treated cells. Ang-II-induced VSMC dedifferentiation, proliferation, migration, KLF4 expression, H2O2 production and the phosphorylation of AKT, ERK1/2 were all inhibited by knockdown of P22phox. Furthermore, H2O2 treatment effectively enhanced the phosphorylation of AKT, ERK1/2 and the expression of KLF4, whereas LY294002 (a specific inhibitor of PI3K), U0126 (a specific inhibitor of ERK1/2) significantly attenuated the H2O2-induced up-regulation of KLF4. In conclusion, these results demonstrated that p22phox promotes Ang-II-induced VSMC phenotypic switch via the H2O2-ERK1/2/AKT-KLF4 signaling pathway.

Cupriavidus sp. strain Cd02-mediated pH increase favoring bioprecipitation of Cd2+ in medium and reduction of cadmium bioavailability in paddy soil.

This study investigated the effects of Cupriavidus sp. strain Cd02-mediated increase on biosorption and bioprecipitation of Cd2+ during the 144-h cultivation time as well as evaluated effectivenesses of changing soil pH and bioavailability of cadmium after bioaugmentation of strain Cd02 into Cd-contaminated paddy soil for 15 days. Results showed that strain Cd02-induced pH increase of the culture medium (from 7.40 to 8.68) facilitated biosorption of Cd2+ on Cd02 cell surface (4.82 mg/mg) and extracellular bioprecipitation in form of cadmium carbonate (3.07 mg/mg). Also, the pH values of Cd-contaminated paddy soil increased by 1.41 units after strain Cd02 was applied for 15 days, which thereby promoted the decrease of exchangeable fraction of Cd2+ by 6.5% in the tested paddy soil. Meanwhile, strain Cd02 could prosperously live in paddy soils after bioaugmentation. These results suggest that strain Cd02 may be applicable for bioremediation of the heavy metal-contaminated soils by bioaugmentation.

Intrapersonal and interpersonal facilitators of forgiveness following spousal infidelity: A stress and coping perspective.

OBJECTIVE: Forgiveness includes processes that involve a decision to stop bitterness and thoughts of revenge (i.e., decisional forgiveness), which further motivates the forgiver towards the restoration of positive emotions (i.e., emotional forgiveness). Using stress and coping framework, this study investigated intrapersonal and interpersonal facilitators of decisional and emotional forgiveness in a Chinese marital context. METHOD: Participants were 154 respondents who had experienced or were experiencing spousal infidelity. RESULTS: Solidarity-oriented personality and perceived partner's reconciliation motivation facilitated benign attributions and empathy, then facilitated higher levels of decisional forgiveness, which promoted emotional forgiveness. Strength of marital bond before the infidelity directly predicted higher levels of emotional forgiveness. CONCLUSIONS: Our findings provide evidence for the differentiated decisional and emotional forgiveness processes after spousal infidelity and delineate different coping mechanism that triggers them, thus lending culturally appropriate evidence for clinicians who work with clients facing spousal infidelity.

Low vascularity predicts favourable outcomes in leiomyoma patients treated with uterine artery embolization.

OBJECTIVES: To investigate the clinical factors predicting outcomes of leiomyoma treated with uterine artery embolization (UAE). METHODS: A total of 183 uterine leiomyoma patients undergoing UAE were retrospectively analyzed. Patient age, characteristics of vascular supply in magnetic resonance imaging (MRI)/digital subtraction angiography (DSA), number, size and location of leiomyoma were recorded. Leiomyoma regrowth, new leiomyoma appearance and recurrence of any previously reported symptoms were carefully monitored over a mean follow-up of 30 months (median 32 months, range 12-80). Potential recurrence risk factors were analyzed by univariate and multivariate cox regression analysis. RESULTS: Twenty-three recurrences were recorded. The difference in the vascularity classification systems between MRI and DSA was not statistically significant (P = 0.059). High vascularity in MRI, high vascularity in DSA and multiple leiomyoma showed a significant risk of recurrence using univariate and multivariate analysis (P = 0.004, P < 0.001 and P = 0.023, respectively). The other factors were not significantly associated with leiomyoma recurrence (P > 0.05). CONCLUSION: Low vascularity and solitary leiomyoma indicated favourable outcomes in patients treated with UAE. KEY POINTS: • Low vascularity and solitary mass predicted favourable outcomes in UAE-treated patients. • MRI might provide information on vascularity in leiomyoma before UAE. • Variations in vascular supply, age, size, location were not associated with recurrence.

[Research progress in the regulation of endothelial nitric oxide synthase activity and cardiovascular diseases].

Endothelial nitric oxide synthase (eNOS) is a major enzyme responsible for nitric oxide (NO) production. Both eNOS and NO play an important role in regulating vessel structure and function through participating in the physiological and pathological process. Generally, the phosphorylation of eNOS has been recognized as a common mechanism for regulation of eNOS function. However, recent studies have demonstrated several other new mechanisms, such as acetylation, S-glutathionylation, and protein-protein interaction. This review highlights the basic structure and functions of eNOS as well as the relationship between regulation of eNOS activity and cardiovascular diseases.

Lithology and niche habitat have significant effect on arbuscular mycorrhizal fungal abundance and their interspecific interactions.

The chemical properties of bedrock play a crucial role in shaping the communities of soil and root-associated arbuscular mycorrhizal fungi (AMF). We investigate AMF community composition and diversity in bulk soil, rhizosphere soil, and roots in karst and non-karst forests. Chemical properties of bedrock of the calcium oxide (CaO) and ratio of calcium oxide and magnesium oxide (Ca/Mg), soil pH, and exchangeable Ca2+ were higher in karst carbonate rocks compared to non-karst clastic rocks. Conversely, bedrock phosphorus content (P-rock), silicon dioxide (SiO2) content, and tree diversity exhibited an opposing trend. AMF abundance was higher in non-karst clastic rocks than in karst carbonate rocks. Stronger interspecific interactions among AMF taxa occurred in the bulk soil and rhizosphere soil of non-karst clastic rocks compared to karst carbonate rocks. AMF abundance and diversity were higher in rhizosphere soil and roots, attributed to increasing nutrient availability when compared to the bulk soil. A more complex network within AMF taxa was observed in rhizosphere soil and roots compared to bulk soil due to an increase in AMF abundance and diversity in rhizosphere soil and roots. Comparing non-karst clastic rocks, karst carbonate rocks increased soil nitrogen (N) and P levels, which can be attributed to the elevated content of soil Ca2+ and Mg2+ content, facilitated by the high CaO content and Ca/Mg ratio in the bedrock of karst forests. However, the thicker soil layer exhibited higher soil nutrient storage, resulting in greater tree diversity in non-karst forests. These findings suggest that high tree richness may increase root biomass and secretion of root exudates in non-karst regions, thereby enhancing the abundance of AMF and their interspecies interactions. Consequently, the diverse bedrock properties that drive variations in soil properties, nutrients, and plant diversity can impact AMF communities, ultimately promoting plant growth and contributing to vegetation recovery.

Supramolecular channels via crown ether functionalized polyaniline for proton-self-doped cathode in aqueous zinc-ion battery.

Polyaniline (PANI) has been widely used as a cathode in aqueous zinc-ion batteries (AZIBs) because of its attractive conductivity and energy storage capability. However, the extensive application of PANI is limited by spontaneous deprotonation and slow diffusion kinetics. Herein, an 18-crown-6-functionalised PANI pseudorotaxane (18C6@PANI) cathode is successfully developed through a facile template-directed polymerisation reaction. The 18C6@PANI cathode exhibits a high specific capacity of 256 mAh g-1 at 0.2 A/g, excellent rate performance of 134 mAh g-1 at 6 A/g and outstanding cycle stability at a high current density of 3 A/g over 10,000 cycles. Experimental and theoretical analyses demonstrate the formation of the -N-Zn-O- structure. The abundant supramolecular channels in pseudorotaxane, induced by crown ether functional groups, are beneficial for achieving superior cyclability and rate capability. These encouraging results highlight the potential for designing more efficient PANI-based cathodes for high-performance AZIBs.

Parental psychological control and children's self-esteem: A longitudinal investigation in children with and without oppositional defiant problems.

Oppositional defiant problems are among the most prevalent psychological problems among children and adolescents from China and across the world. Still little is understood about how self-esteem, in conjunction with parenting experiences, develops in children with oppositional defiant problems. We addressed this gap of knowledge in a two-year longitudinal study. Specifically, we explored how parental psychological control predicts children's self-esteem levels over time, and in turn, how children's self-esteem levels predict parental psychological control. We collected data in Chinese children (ages 8 to 13 at T1) with (N = 224) and without (N = 217) oppositional defiant problems, and tested three-wave cross-lagged panel models. Multigroup analyses showed that the associations between parental psychological control and children's self-esteem were the same for children with and without oppositional defiant problems. Results for the total sample revealed bi-directional associations between maternal psychological control and children's self-esteem. Children who perceived more psychological control from their mothers were likely to exhibit lower self-esteem over time, and vice versa, children with lower self-esteem were likely to perceive more maternal psychological control over time. Conversely, a unidirectional paternal effect was observed in father-child dyads. Our findings help understand the parent-child dynamics that shape the psychological development of children with oppositional defiant problems.

Comparative Study of 5-Aminolevulinic Acid-Mediated Photodynamic Therapy and the Loop Electrosurgical Excision Procedure for the Treatment of Cervical High-Grade Squamous Intraepithelial Lesions.

The loop electrosurgical excision procedure (LEEP) is a common treatment for cervical intraepithelial neoplasia (CIN). Photodynamic therapy (PDT) mediated by 5-aminolevulinic acid (ALA) is a non-invasive modality that has been used for treating precancerous diseases and HPV infections. This comparative study evaluated the efficacy and safety of ALA PDT and the LEEP in the treatment of cervical high-grade squamous intraepithelial lesions (HSILs). Patient records were reviewed and HSIL patients with HPV infections (24-51 years old) who underwent PDT or LEEP treatment were selected. Efficacy was analyzed blindly based on HPV-DNA, cytology, and colposcopy-directed biopsy obtained at 6 months after treatment. Treatment-related discomfort and side effects were also analyzed. Cure rates of 88.1% and 70.0% were achieved for the PDT group and LEEP group (p < 0.05), respectively. HPV-negative conversion rates of 81.0% and 62.0% were achieved for the PDT group and LEEP group (p < 0.05), respectively. The overall lesion remission rate of the PDT group was 19% higher than that of the LEEP group. The incidence of side effects was much lower in the PDT group. These results show that ALA PDT is a feasible non-invasive treatment for cervical HSIL.

Ultrasensitive one-pot detection of monkeypox virus with RPA and CRISPR in a sucrose-aided multiphase aqueous system.

IMPORTANCE: The monkeypox virus was declared as a Public Health Emergency of International Concern (PHEIC) by the World Health Organization (WHO) and continues to cause infection cases worldwide. Given the risk of virus evolution, it is essential to identify monkeypox virus infection in a timely manner to prevent outbreaks. This study establishes a novel one-pot recombinase polymerase amplification-Clustered Regularly Interspaced Short Palindromic Repeats (RPA-CRISPR) assay for monkeypox virus with an ultra-high sensitivity. The assay shows good specificity, accuracy, and the rapidness and convenience important for point-of-care testing. It provides an effective tool for the early diagnosis of monkeypox, which is useful for the prevention of an epidemic.

Therapeutic effect of ZnO NPs-polyhexanide-hydrogel on Staphylococcus aureus-induced skin wound infection in mice.

Nanometer zinc oxide (ZnONPs) offers strong antibacterial, wound healing, hemostatic benefits, and UV protection. Additionally, poly(hexamethylene biguanide)hydrochloride (PHMB) is an environmentally friendly polymer with strong bactericidal properties. However, the synergistic effect of the combination of ZnONPs and PHMB has not been previously explored. The purpose of this study is to explore the synergies of ZnONPs and PHMB and the healing efficacy of ZnO NPs-PHMB-hydrogel on skin wounds in mice infected with Staphylococcus aureus. Therefore, the mice were subjected to skin trauma to create a wound model and were subsequently infected with S. aureus, and then divided into various experimental groups. The repair effect was evaluated by assessing the healing rate, as well as measuring the levels of TNF-α, IL-2, EGF, and TGF-ß1 contents in the tissue. On the 4th and 9th days post-modeling, the Z-P group exhibited notably higher healing rates compared to the control group. However, on the 15th day, both the Z-P and AC groups achieved healing rates exceeding 99%. ZnO NPs-PHMB-hydrogel promoted the formation of a fully restored epithelium, increased new hair follicles and sebaceous glands beneath the epidermis, and markedly reduced inflammatory cell infiltration, which was markedly distinct from the control group. On the 7th day, the Z-P group exhibited significantly higher levels of EGF and TGF-ß1, along with a considerable reduction in the TNF-α levels as compared with the control group. These results affirmed that ZnO NPs-PHMB-hydrogel effectively inhibits S. aureus infection and accelerates skin wound healing.