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Abstract Background CRISPR-Cas13a is renowned for its precise and potent RNA editing capabilities in cancer therapy. While various material systems have demonstrated efficacy in supporting CRISPR-Cas13a to execute cellular functions in vitro efficiently and specifically, the development of CRISPR-Cas13a-based therapeutic agents for intravesical instillation in bladder cancer (BCa) remains unexplored. Methods In this study, we introduce a CRISPR-Cas13a nanoplatform, which effectively inhibits PDL1 expression following intravesical instillation. This system utilizes a fusion protein CAST, created through the genetic fusion of CRISPR-Cas13 and the transmembrane peptide TAT. CAST acts as a potent transmembrane RNA editor and is assembled with the transepithelial delivery carrier fluorinated chitosan (FCS). Upon intravesical administration into the bladder, the CAST-crRNAa/FCS nanoparticles (NPs) exhibit remarkable transepithelial capabilities, significantly suppressing PDL1 expression in tumor tissues.To augment immune activation within the tumor microenvironment, we integrated a fenbendazole (FBZ) intravesical system (FBZ@BSA/FCS NPs). This system is formulated through BSA encapsulation followed by FCS coating, positioning FBZ as a powerful chemo-immunological agent. Results In an orthotropic BCa model, the FBZ@BSA/FCS NPs demonstrated pronounced tumor cell apoptosis, synergistically reduced PDL1 expression, and restructured the immune microenvironment. This culminated in an enhanced synergistic intravesical instillation approach for BCa. Consequently, our study unveils a novel RNA editor nanoagent formulation and proposes a potential synergistic therapeutic strategy. This approach significantly bolsters therapeutic efficacy, holding promise for the clinical translation of CRISPR-Cas13-based cancer perfusion treatments.

Abstract Rheumatoid arthritis (RA) involves chronic joint inflammation. Combining acupuncture and medication for RA treatment faces challenges like spatiotemporal variability, limited drug loading in acupuncture needles, and premature or untargeted drug release. Here, we designed a new type of tubular acupuncture needles, with an etched hollow honeycomb-like structure to enable the high loading of therapeutics, integrating the traditional acupuncture and drug repository into an all-in-one therapeutic platform. In these proof-of-concept experiments, we fabricated injectable hollow honeycomb electroacupuncture needles (HC-EA) loaded with melittin hydrogel (MLT-Gel), enabling the combination treatment of acupuncture stimulation and melittin therapy in a spatiotemporally synchronous manner. Since the RA microenvironment is mildly acidic, the acid-responsive chitosan (CS)/sodium beta-glycerophosphate (β-GP)/ hyaluronic acid (HA) composited hydrogel (CS/GP/HA) was utilized to perform acupuncture stimulation and achieve the targeted release of injected therapeutics into the specific lesion site. Testing our therapeutic platform involved a mouse model of RA and bioinformatics analysis. MLT-Gel@HC-EA treatment restored Th17/Treg-mediated immunity balance, reduced inflammatory factor release (TNF-α, IL-6, IL-1β), and alleviated inflammation at the lesion site. This novel combination of modified acupuncture needle and medication, specifically melittin hydrogel, holds promise as a therapeutic strategy for RA treatment. Graphical abstract

Oridonin (ORI), an ent-kaurane diterpenoid derived from Rabdosia rubescens (Hemsl.) H.Hara, serves as the primary bioactive component of this plant. It demonstrates a broad spectrum of therapeutic activities, including moderate to potent anticancer properties, alongside anti-inflammatory, antibacterial, antifibrotic, immunomodulatory, and neuromodulatory effects, thus influencing diverse biological processes. However, its clinical potential is significantly constrained by poor aqueous solubility and limited bioavailability. In alignment with the approach of developing drug candidates from natural compounds, various strategies, such as structural modification and nanocarrier systems, have been employed to address these challenges. This review provides an overview of ORI-based nano-delivery systems, emphasizing their potential to improve the clinical applicability of oridonin in oncology. Although some progress has been made in advancing ORI nano-delivery research, it remains insufficient for clinical implementation, necessitating further investigation.

The emergence of myelinating oligodendrocytes represents a pivotal developmental milestone in vertebrates, given their capacity to ensheath axons and facilitate the swift conduction of action potentials. It is widely accepted that cortical oligodendrocyte progenitor cells (OPCs) arise from medial ganglionic eminence (MGE), lateral/caudal ganglionic eminence (LGE/CGE), and cortical radial glial cells (RGCs). Here, we used two different fate mapping strategies to challenge the established notion that the LGE generates cortical OPCs. Furthermore, we used a Cre/loxP-dependent exclusion strategy to reveal that the LGE/CGE does not give rise to cortical OPCs. Additionally, we showed that specifically eliminating MGE-derived OPCs leads to a significant reduction of cortical OPCs. Together, our findings indicate that the LGE does not generate cortical OPCs, contrary to previous beliefs. These findings provide a new view of the developmental origins of cortical OPCs and a valuable foundation for future research on both normal development and oligodendrocyte-related disease.

Operation of short food supply chains (abbr. SFSCs) during the period of the pandemic caused by the SARS-CoV-2 virus provided a lesson which might be useful for perceiving the behaviour of farmers and local food systems in potential future crises caused by various endogenous or exogenous factors. A study based on a sample of 1081 beekeepers in Western Balkan countries shows how beekeepers involved in SFSCs perceived the influence of the pandemic on the growth of demand and sale of honey through these marketing channels. On the scale from 1 (no influence) to 7 (very strong influence), the average rating of 3.53 indicated a weak to moderate influence, which was fairly consistent across the countries of this region. The Kruskal-Wallis test and Mann-Whitney test determined that large-scale beekeepers, those in mountainous areas, those adding value to honey and beekeepers producing other bee products apart from honey stated that the pandemic had a stronger influence on the growth of demand and sale of honey through SFSCs. At the same time, a higher rating of the pandemic’s influence on sales growth was accompanied by greater limitations of beekeepers regarding the availability of family labour for selling honey, as well as by greater needs for digital marketing knowledge and skills. Binary logistic regression showed that the increase in beekeepers’ age led to the decreased rating of the influence of the pandemic on the growth of demand and sale of honey through SFSCs. It also showed that the influence rating rose with the increase in production capacity, adding value to honey and beekeepers’ needs for digital marketing knowledge and skills. The obtained results provide valuable knowledge about the potential response of SFSCs and involved producers to future crises and disruptions. It is primarily intended for policy makers, but also to practitioners and scientific and expert communities. All of them should respond proactively on behalf of society and prepare themselves for future challenges.

Porcine parvoviruses one through eight (PPV1-PPV8) are prevalent in Chinese swine herds. However, the infection status of all these PPVs in slaughtered pigs is still unclarified. In this study, we detected PPV1-PPV8 in 353 tissue samples collected from slaughtered pigs from six regions of China in 2023. At least one species of PPV was detected in 79.32% of the samples (280 out of 353). Six PPV species were detected, except for PPV4 and PPV8, in slaughtered pigs, within which PPV3 (49.86%), PPV2 (42.49%), and PPV7 (42.21%) were predominant, followed by PPV1 (13.31%), PPV6 (13.31%), and PPV5 (8.22%). Noticeably, co-infection was frequently detected, with 67.50% of PPV-positive samples (189 out of 280) co-infecting with two to six PPVs. In addition, one representative genome for each detected PPV was determined. Multiple sequence alignment determined a large number of substitutions in capsid proteins of PPVs. Genome-based phylogenetic analysis confirmed the PCR detection results. Recombination detection identified two potential recombinants (PPV2 GDCZ2023-2088 strain and PPV3 HLJSYS2023-1654 strain) in slaughtered pigs. Overall, this study provides new insights into the prevalence and evolution of PPVs, particularly in slaughtered pigs in China.

Introduction Mental health-related stigma occurs not only within the public community but is also an issue among healthcare professionals. The relationship between national culture and provider stigma remains yet to be empirically attested. Objectives We performed a cross-sectional multicentre study across 32 European countries to investigate the attitudes of psychiatrists towards patients with mental health problems. We aimed to examine the relationship of attitude with country-specific indicators. Methods We measured stigmatizing attitudes using the Opening Minds Stigma Scale for Health Care Providers (OMS-HC) within an online survey among specialists and trainees in general adult, child and adolescent psychiatry. Its total score was correlated with the Human Development Index (HDI), the Democracy Index (DI), the Social Progress Index (SPI), the number of psychiatrists per 100,000 people, and the Hofstede dimensions. Latent class analysis was done to find subgroups of countries according to the stigmatizing attitudes of psychiatrists and the six Hofstede dimensions. Results Altogether, n=4245 participants completed the survey. The total score of the OMS-HC significantly correlated with the long-term orientation (r=0.453, p=0.015) and indulgence dimensions (r=-0.629, p

BackgroundDuring the use of electronic cigarettes (e-cigarettes), users are still exposed to carcinogens similar to those found in tobacco products. Since these carcinogens are metabolized and excreted in urine, they may have carcinogenic effects on the bladder urinary tract epithelium. This meta-analysis aimed to compare bladder cancer carcinogens in the urine of tobacco users and e-cigarette users using a large number of samples.MethodsA systematic meta-analysis was performed using data obtained from several scientific databases (up to November 2023). This cumulative analysis was performed following the Preferred Reporting Items for Systematic Evaluation and Meta-Analysis (PRISMA) and Assessing the Methodological Quality of Systematic Evaluations (AMSTAR) guidelines, according to a protocol registered with PROSPERO. This study was registered on PROSPERO and obtained the unique number: CRD42023455600.ResultsThe analysis included 10 high-quality studies that considered polycyclic aromatic hydrocarbons (PAHs), volatile organic compounds (VOCs) and tobacco-specific nitrosamines (TSNAs). Statistical indicators show that there is a difference between the tobacco user group and the e-cigarette user group in terms of 1-Hydroxynaphthalene (1-NAP) [weighted mean difference (WMD)10.14, 95% confidence interval (CI) (8.41 to 11.88), p < 0.05], 1-Hydroxyphenanthrene (1-PHE) [WMD 0.08, 95% CI (−0.14 to 0.31), p > 0.05], 1-Hydroxypyrene (1-PYR) [WMD 0.16, 95% CI (0.12 to 0.20), p < 0.05], 2-Hydroxyfluorene (2-FLU) [WMD 0.69, 95% CI (0.58 to 0.80), p < 0.05], 2-Hydroxynaphthalene (2-NAP) [WMD 7.48, 95% CI (4.15 to 10.80), p < 0.05], 3-Hydroxyfluorene (3-FLU) [WMD 0.57, 95% CI (0.48 to 0.66), p < 0.05], 2-Carbamoylethylmercapturic acid (AAMA) [WMD 66.47, 95% CI (27.49 to 105.46), p < 0.05], 4-Hydroxy-2-buten-1-yl-mercapturic acid (MHBMA) [WMD 287.79, 95% CI (−54.47 to 630.04), p > 0.05], 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNAL) [WMD 189.37, 95% CI (78.45 to 300.29), p < 0.05], or N0-nitrosonornicotine (NNN) [WMD 11.66, 95% CI (7.32 to 16.00), p < 0.05].ConclusionUrinary bladder cancer markers were significantly higher in traditional tobacco users than in e-cigarette users.Systematic review registration: PROSPERO (CRD42023455600: https://www.crd.york.ac.uk/PROSPERO/).

Low-temperature bonding of nanotwinned (NT-) metals holds great potential for enhancing the performance and reliability of electronic interconnections. In this study, we performed low-temperature bonding of (111)-oriented nanotwinned silver (NT-Ag) films and thoroughly investigated the influence of (111) texture on the interfacial microstructure and bonding strength of the joints. The results demonstrated that high-quality bonding of highly (111)-oriented NT-Ag films was achieved even at 200 °C in air. The defect-free, coherent Ag(111)Ag(111) bonding interface was formed via face-to-face bonding of the highly (111)-oriented columnar grains. The shear strength of the joints significantly increased with the rise in (111) texture intensity. Moreover, we developed the void closure model to quantitatively elucidate the contribution of (111) texture to void closure and bonding ratio. Modeling results revealed that the (111) texture, with the highest surface diffusion coefficient, greatly increased the interfacial bonding ratio by accelerating surface source diffusion, thereby enabling high-strength bonding at low temperatures. The experimental validation and modeling results presented here contribute a novel perspective for optimizing interfacial microstructure and bonding strength by regulating the surface texture in the solid-state bonding of metals.

Ultrasound-assisted Kovar/SnSb10 assemblies are promising candidates for high-precision electronic devices; however, limited research has been conducted on the effect of ultrasonication on its interfacial evolution and mechanical properties. In this study, the microstructure evolution, orientation relationships (ORs), interinhibitive dissolution-precipitation mechanism, and mechanical properties of a Kovar/SnSb10 assembly with/without ultrasonication were systematically investigated. The morphology of the intermetallics (IMCs) evolved from a newly-formed, lash-type to a block-type morphology with an increasing reflow duration, and dentation-type IMCs appeared after ultrasonication. The interfacial IMC without ultrasonication mainly contained FeSn2 and a minority of Ni3Sn2, whereas the strong disturbance caused by ultrasound led to the formation of FeSn in the peripheral FeSn3 and NiSn IMCs. Furthermore, high-resolution transmission electron microscopy (HRTEM) analysis revealed that the ORs of IMC/solder and IMC/Kovar transformed from coherent and semi-coherent to incoherent relationships after ultrasonic treatment. The ORs of the IMC/solder growth front after ultrasonication were [0 1¯ 1]Sn∥[1 1 0]FeSn3, (0 1 1)Sn∥(0 0 1¯)FeSn3 and [0 1¯ 1]Sn∥[5¯ 1 0]NiSn, (0 1 1)Sn∥(0 1¯ 0)NiSn. Additionally, 60 s ultrasonic treatment promoted the growth of IMCs, and increased the ultimate shear strength by approximately 20 MPa, despite the emergence of incoherent interfaces. This phenomenon was primarily attributed to the dispersion strengthening effect of the abundant ultrasound-induced nano-Ni3Sn2 precipitates. This study substantiates the feasibility of ultrasonication-assisted Kovar/SnSb10 assemblies and introduces a novel concept for low-cost, high-efficiency, and high-performance packaging.

In the genesis and progression of cardiovascular diseases involving both innate and adaptive immune responses, inflammation plays a pivotal and dual role. Studies in experimental animals indicate that certain immune responses are protective, while others exacerbate the disease. T-helper (Th) 1 cell immune responses are recognized as key drivers of inflammatory progression in cardiovascular diseases. Consequently, the CD4+CD25+FOXP3+ regulatory T cells (Tregs) are gaining increasing attention for their roles in inflammation and immune regulation. Given the critical role of Tregs in maintaining immune-inflammatory balance and homeostasis, abnormalities in their generation or function might lead to aberrant immune responses, thereby initiating pathological changes. Numerous preclinical studies and clinical trials have unveiled the central role of Tregs in cardiovascular diseases, such as atherosclerosis. Here, we review the roles and mechanisms of Treg subsets in cardiovascular conditions like atherosclerosis, hypertension, myocardial infarction and remodeling, myocarditis, dilated cardiomyopathy, and heart failure. While the precise molecular mechanisms of Tregs in cardiac protection remain elusive, therapeutic strategies targeting Tregs present a promising new direction for the prevention and treatment of cardiovascular diseases.

BackgroundPrevious observational researchers have found an inverse bidirectional link between Alzheimer’s disease (AD) and prostate cancer (PCa); yet, the causative nature of this link remains unclear. To investigate the causal interactions between AD and PCa, a bidirectional Mendelian randomization (MR) analysis was conducted.MethodsThis study comprised two Genome-Wide Association Study (GWAS) summary statistics for AD (17,008 cases and 37,154 controls) and PCa (79,148 cases and 61,106 controls) in individuals of European ancestry. The inverse-variance weighted (IVW) method was employed as the primary approach, while MR-Egger, weighted median, weighted mode, and simple mode served as supplementary methods for estimating the causal effect. To assess pleiotropy, the MR-PRESSO global test and MR-Egger regression were used. Cochran’s Q test was adopted to check heterogeneity, MR Steiger test and the leave-one-out analysis was performed to confirm the robustness and reliability of the results.ResultsThe causal association genetically inferred of AD on PCa was found using IVW (OR = 0.974, 95% CI = 0.958-0.991, p = 0.003) in forward MR analysis and the causal association genetically inferred of PCa on AD was not found using IVW (OR = 1.000, 95% CI: 0.954-1.049, P = 0.988) in reverse MR analysis. The sensitivity analysis showed that no pleiotropy and heterogeneity was observed. The leave-one-out analysis showed that the findings were not inordinately affected by any instrumental variables.ConclusionThe results of this study demonstrated an absence of bidirectional causality between AD and PCa among the European population, suggested that a genetically predicted possibility of decreased PCa risk in AD patients, and no significant genetically predicted causal effect of PCa on AD.

BackgroundAspirin, as one of the most commonly used drugs, possesses a broad spectrum of therapeutic applications. Presently, the potential association between aspirin usage and the risk elevation of erectile dysfunction (ED) remains inconclusive. The objective of this study employing two-sample Mendelian randomization (MR) was to clarify the causal impact of aspirin use on the risk of ED.MethodsThis study incorporated two sets of Genome-Wide Association Study (GWAS) summary statistics, one for aspirin use (46,946 cases and 286,635 controls) and another for ED (6,175 cases and 217,630 controls) in individuals of European ancestry. The inverse-variance weighted (IVW) method was employed as the primary approach, supplemented by MR-Egger, weighted median, weighted mode, and simple mode to estimate the causal effect of aspirin usage on the risk of ED development. To assess pleiotropy, the MR-PRESSO global test and MR-Egger regression were used. Cochran’s Q test was adopted to check heterogeneity, and the leave-one-out analysis was performed to confirm the robustness and reliability of the results.ResultsThe causal association between genetically inferred aspirin use and ED was found by using inverse variance weighted (OR = 20.896, 95% confidence interval = 2.077-2.102E+2, P = 0.010). The sensitivity analysis showed that no pleiotropy and heterogeneity was observed. Furthermore, the leave-one-out analysis demonstrated that the findings were not significantly affected by any instrumental variables.ConclusionThe results of this study highlighted the significance of aspirin use as a predisposing factor for ED and provided further evidence supporting the causal association between aspirin utilization and ED within European populations.

Introduction: Lipoma is a common benign, slow-growing soft tissue neoplasm. Periosteal lipomas of the proximal radius causing posterior interosseus nerve (PIN) palsy are the rarest. Due to specific anatomical relationships, proximal antebrachial lipomas can easily compress PIN. We present a case report and report on the related literature. Material and Methods: A 55-year-old female was admitted complaining of a progressively growing lump in the anterior-lateral antebrachial region near the left elbow. The recent onset of weakness in finger extension was more evident during manual work. MRI showed a well-defined oval lesion in the dorsal aspect of the proximal radius bone with inter-muscular laying between m. supinator et extensor carpi radialis brevis of 7x5x3 cm in diameter. EMG study confirmed PIN compression syndrome. An en-block extirpation was performed through extensor muscles. PIN and its muscular branches were well preserved. Histological examination confirmed lipoma. The postoperative course was uneventful, and a good recovery was seen within 12 days. We revised three other similar cases operated for the same Clinical diagnosis by our team. Pertinent literature was reported. Discussion: Periosteal radius lipoma related to compression of PIN has rarely been reported in the literature. We say our operative series and the surgical technique. MRI and EMG are the standard diagnostic methods. The intermuscular approach is safe for total tumor removal in experienced hands. Conclusion: Periosteal lipoma compressing PIN is a rare clinical finding. Total removal may be obtained through an intermuscular approach. Intraoperative monitoring can assist in preserving tiny PIN branches

Background: Monitoring SARS-CoV-2 seroprevalence dynamics during the COVID-19 pandemic is crucial for understanding population immunity and providing insights into public health policies. Limited data exist on this from Albania and other Eastern European countries. This study aimed to investigate SARS-CoV-2 seroprevalence in Albania, comparing August 2021 and August 2022 data from two representative samples of the general population. The objective was to understand the temporal dynamics of SARS-CoV-2 antibodies across age groups and assess the impacts of natural infection and vaccination on population immunity. Methods: This longitudinal study was conducted in two consecutive cross-sectional assessments 12 months apart in Albania's urban all-ages population. IgG anti-Spike-1 and anti-Nucleoprotein SARS-CoV-2 antibodies were measured using ELISA, focusing on seropositivity rates and antibody levels. Methods: The study encompassed 2143 and 2183 individuals in August 2021 and 2022, respectively, with the anti-S1-IgG seropositivity rate escalating from 70.9 ​% to 92.1 ​%. In 2021, seroprevalence ranged from 49.6 ​% (0–15 years) to 82 ​% (>60 years). By August 2022, it surpassed 90 ​% in most age groups, except 0–15 years (73.8 ​%). ''Hybrid'' immunity (COVID-19+ and Vaccine+) reached 56.6 ​% in 2022, or 2.8 times higher than in 2021, exhibiting the highest antibody levels compared to the only vaccinated or previously COVID-19-infected individuals. Conclusion: This study highlights an overall 94 % seroprevalence in the Albanian population in August 2022 and robust ''hybrid'' immunity, suggesting substantial protective immunity against SARS-CoV-2. The lower immunity in the 0–15 age group underscores the necessity for youth-targeted vaccine campaigns. These findings provide valuable insights for shaping healthcare measures and vaccination policies.