Enantioselective toxicity of the neonicotinoid dinotefuran on honeybee (Apis mellifera) larvae.

The threat of neonicotinoids to insect pollinators, particularly honeybees (Apis mellifera), is a global concern, but the risk of chiral neonicotinoids to insect larvae remains poorly understood. In the current study, we evaluated the acute and chronic toxicity of dinotefuran enantiomers to honeybee larvae in vitro and explored the mechanism of toxicity. The results showed that the acute median lethal dose (LD50) of S-dinotefuran to honeybee larvae was 30.0 µg/larva after oral exposure for 72 h, which was more toxic than rac-dinotefuran (92.7 µg/larva) and R-dinotefuran (183.6 µg/larva). Although the acute toxicity of the three forms of dinotefuran to larvae was lower than that to adults, chronic exposure significantly reduced larval survival, larval weight, and weight of newly emerged adults. Analysis of gene expression and hormone titer indicated that dinotefuran affects larval growth and development by interfering with nutrient digestion and absorption and the molting system. Analysis of hemolymph metabolome further revealed that disturbances in the neuroactive ligand-receptor interaction pathway and energy metabolism are the key mechanisms of dinotefuran toxicity to bee larvae. In addition, melatonin and vitellogenin are used by larvae to cope with dinotefuran-induced oxidative stress. Our results contribute to a comprehensive understanding of dinotefuran damage to bees and provide new insights into the mechanism of enantioselective toxicity of insecticides to insect larvae.

Stem Cell-Derived Nanovesicles Embedded in Dual-Layered Hydrogel for Programmed ROS Regulation and Comprehensive Tissue Regeneration in Burn Wound Healing.

Burn wounds often bring high risks of delayed healing process and even death. Reactive oxygen species (ROS) play a crucial role in burn wound repair. However, the dynamic process in wound healing requires both the generation of ROS to inhibit bacteria and the subsequent reduction of ROS levels to initiate and promote tissue regeneration, which calls for a more intelligent ROS regulation dressing system. Hence, a dual-layered hydrogel (Dual-Gel) tailored to the process of burn wound repair is designed: the inner layer hydrogel (Gel 2) first responds to bacterial hyaluronidase (Hyal) to deliver aggregation-induced emission photosensitizer functionalized adipose-derived stem cell nanovesicles, which generate ROS upon light irradiation to eliminate bacteria; then the outer layer hydrogel (Gel 1) continuously starts a long-lasting consumption of excess ROS at the wound site to accelerate tissue regeneration. Simultaneously, the stem cell nanovesicles trapped in the burns wound also provide nutrients and mobilize neighboring tissues to thoroughly assist in inflammation regulation, cell proliferation, migration, and angiogenesis. In summary, this study develops an intelligent treatment approach on burn wounds by programmatically regulating ROS and facilitating comprehensive wound tissue repair.

Toxic effects of acaricide fenazaquin on development, hemolymph metabolome, and gut microbiome of honeybee (Apis mellifera) larvae.

Fenazaquin, a potent insecticide widely used to control phytophagous mites, has recently emerged as a potential solution for managing Varroa destructor mites in honeybees. However, the comprehensive impact of fenazaquin on honeybee health remains insufficiently understood. Our current study investigated the acute and chronic toxicity of fenazaquin to honeybee larvae, along with its influence on larval hemolymph metabolism and gut microbiota. Results showed that the acute median lethal dose (LD50) of fenazaquin for honeybee larvae was 1.786 µg/larva, and the chronic LD50 was 1.213 µg/larva. Although chronic exposure to low doses of fenazaquin exhibited no significant effect on larval development, increasing doses of fenazaquin resulted in significant increases in larval mortality, developmental time, and deformity rates. At the metabolic level, high doses of fenazaquin inhibited nucleotide, purine, and lipid metabolism pathways in the larval hemolymph, leading to energy metabolism disorders and physiological dysfunction. Furthermore, high doses of fenazaquin reduced gut microbial diversity and abundance, characterized by decreased relative abundance of functional gut bacterium Lactobacillus kunkeei and increased pathogenic bacterium Melissococcus plutonius. The disrupted gut microbiota, combined with the observed gut tissue damage, could potentially impair food digestion and nutrient absorption in the larvae. Our results provide valuable insights into the complex and diverse effects of fenazaquin on honeybee larvae, establishing an important theoretical basis for applying fenazaquin in beekeeping.

Proteomics profiling of the honeybee parasite Tropilaelaps mercedesae across post-embryonic development.

Tropilaelaps mercedesae, an ectoparasitic mite of honeybees, is currently a severe health risk to Apis mellifera colonies in Asia and a potential threat to the global apiculture industry. However, our understanding of the physiological and developmental regulation of this pest remains significantly insufficient. Using ultra-high resolution mass spectrometry, we provide the first comprehensive proteomic profile of T. mercedesae spanning its entire post-embryonic ontogeny, including protonymphs, deutonymphs, mature adults, and reproductive mites. Consequently, a total of 4,422 T. mercedesae proteins were identified, of which 2,189 proteins were significantly differentially expressed (FDR < 0.05) throughout development and maturation. Our proteomic data provide an important resource for understanding the biology of T. mercedesae, and will contribute to further research and effective control of this devastating honeybee pest.

Virulence and Adhesion of the Treponema pallidum Nichols Strain Simultaneously Decrease in a Continuous-Infection New Zealand White Rabbit Model.

Syphilis is a sexually transmitted disease caused by T. pallidum, and the T. pallidum Nichols strain is widely used with the New Zealand white rabbit model for evaluating drug and vaccine protection. However, changes in the virulence of T. pallidum during transmission are still unknown. Herein, we explored the virulence of T. pallidum in the rabbit model of continuous infection through phenotype observation and further investigated the relationship between virulence and adhesion. During the construction of the syphilis rabbit model, the optimal dose of 104/site of T. pallidum was determined to effectively observe the depiction of syphilis lesions and immune responses for further virulence evaluation. Its virulence was gradually weakened during the interaction with host cells or the testicular passage, which was also proven using the pathological phenotype of the syphilis rabbit model. In addition, the adhesive ability of T. pallidum was reduced with increasing generation, which was verified via the co-incubation of the pathogen with Sf1Ep cells. This study provides insight into the relationship by which the virulence and adhesion of T. pallidum were decreased in a New Zealand white rabbit model of continuous infection and contributes to our knowledge regarding the development of syphilis.

A tunable fluorescent probe for superoxide anion detection during inflammation caused by Treponema pallidum.

Syphilis, caused by Treponema pallidum (T. pallidum), is associated with the oxidative stress due to its inflammation-like symptom, and detecting the reactive oxygen species (ROS) is crucial for monitoring the infectious process. Herein, we design and synthesize a perylene-based tunable fluorescent probe, PerqdOH, which can detect endogenous O2˙- during T. pallidum infection. The fluorescence peak shifted from 540 nm to 750 nm with increasing O2˙- levels. Besides, both decreased green fluorescence and enhanced red fluorescence could be observed simultaneously during the in vitro infection, providing the real-time monitoring of intracellular O2˙- caused by T. pallidum. Furthermore, the probe exhibited a remarkable signal in the treponemal lesions on the back of a rabbit model. Taken together, our synthesized PerqdOH holds great potential for application in clarifying the infectious process caused by T. pallidum in real time.

Combination therapy with DHA and BMSCs suppressed podocyte injury and attenuated renal fibrosis by modulating the TGF-ß1/Smad pathway in MN mice.

Artemisinin has immunomodulatory, anti-inflammatory, and antifibrotic effects. Some studies have demonstrated that artemisinins have a protective effect on the kidney. DHA is a derivative of artemisinin and has effects similar to those of artemisinin. Human bone marrow-derived mesenchymal stem cells (BMSCs) accelerate renal repair following acute injury. In the study, we investigated the effects of combination therapy with DHA and BMSCs on membranous nephropathy (MN) mice. The 24-h urinary protein, serum total cholesterol (TC) and triglyceride (TG) levels, and renal histopathology, were measured to evaluate kidney damage. Anti-PLA2R, IgG, and complement 3 (C3) were detected by ELISA. The expression levels of the podocyte injury-related proteins were analyzed by immunohistochemistry. The protein expression levels of α-SMA, ED-1, TGF-ß1, p-Smad2, and p-Smad3 were detected by western blot to analyze renal fibrosis and its regulatory mechanism. Results showed that combination therapy with DHA and BMSCs significantly ameliorated kidney damage in MN model mice by decreasing the levels of 24 h urinary protein, TC and TG. This combination therapy also improved renal histology and reduced the expression of IgG and C3 in the glomerulus. In addition, this combination therapy decreased the expression of podocin and nephrin and relieved renal fibrosis by downregulating α-SMA and ED-1. Furthermore, this combination therapy suppressed TGF-ß1 expression and Smad2/3 phosphorylation. This result (i.e., this combination therapy inhibited the TGF-ß1/Smad pathway) was also supported in vitro. Taken together, combination therapy with DHA and BMSCs ameliorated podocyte injury and renal fibrosis in MN mice by downregulating the TGFß1/Smad pathway.

[Effect of moxibustion on renal function and hypercoagulable state in patients with idiopathic membranous nephropathy of low to medium risk with spleen-kidney deficiency and blood stasis].

OBJECTIVE: To compare the effect of moxibustion combined with basic treatment and simple basic treatment on the clinical symptoms, renal function and hypercoagulable state in patients with idiopathic membranous nephropathy (IMN) of low to medium risk with spleen-kidney deficiency and blood stasis. METHODS: A total of 60 patients with IMN of low to medium risk with spleen-kidney deficiency and blood stasis were randomized into an observation group (30 cases, 2 cases dropped off) and a control group (30 cases, 1 case dropped off). In the control group, the conventional basic treatment of anti-hypertension, regulating blood lipid and anti-coagulation was adopted. On the basis of the control group, moxibustion was applied at Shenshu (BL 23), Pishu (BL 20), Guanyuan (CV 4), Zusanli (ST 36) and Sanyinjiao (SP 6) in the observation group, once a day, 5 days a week continuously with 2 day interval. The treatment of 6 months was required in the both groups. Before treatment and 3 and 6 months into treatment, the total TCM syndrome score, the renal function indexes (24-hour urinary protein quantity [UTP], albumin [ALB], urea nitrogen [BUN] and creatinine [Scr]), the blood coagulation indexes (fibrinogen [FIB], D-Dimer [D-D], p-selection and von Willebrand factor [vWF]), total cholesterol (TC) and triacylglycerol (TG) levels were observed, and the therapeutic efficacy was evaluated on 3 and 6 months into treatment in the two groups. RESULTS: The effective rates of 3 and 6 months into treatment were 78.6% (22/28) and 89.3% (25/28) in the observation group, which were higher than 62.1% (18/29) and 75.9% (22/29) in the control group respectively (P<0.05). On 3 and 6 months into treatment, the total TCM syndrome scores were decreased compared before treatment in the both groups (P<0.05), and those in the observation group were lower than the control group (P<0.05). On 3 months into treatment, the levels of UTP, FIB, D-D, P-selection and vWF were decreased (P<0.05), the level of ALB was increased (P<0.05) compared before treatment in the observation group; the levels of UTP and FIB were decreased compared before treatment in the control group (P<0.05); the level of ALB in the observation group was higher than that in the control group (P<0.05), the levels of FIB and vWF in the observation group were lower than those in the control group (P<0.05). On 6 months into treatment, the levels of UTP, FIB, D-D, P-selection, vWF, TC and TG were decreased (P<0.05), the levels of ALB were increased (P<0.05) compared before treatment in the both groups (P<0.05); the levels of UTP, FIB, D-D, P-selection, vWF, TC and TG in the observation group were lower than those in the control group, the level of ALB in the observation group was higher than that in the control group (P<0.05). CONCLUSION: Moxibustion combined with basic treatment can effectively improve the clinical symptoms, renal function and renal microcirculation in patients with idiopathic membranous nephropathy of low to medium risk with spleen-kidney deficiency and blood stasis, the therapeutic effect is superior to the simple basic treatment.

Circular RNA ABCB10 contributes to laryngeal squamous cell carcinoma (LSCC) progression by modulating the miR-588/CXCR4 axis.

Laryngeal squamous cell carcinoma (LSCC) is a common head and neck cancer with a high metastasis and poor prognosis. Circular RNAs (circRNAs) are a type of non-coding RNAs (ncRNAs) with regulatory function and broadly participate in cancer development. However, the correlation of circular RNA ABCB10 (circABCB10) with LSCC remains unclear. Here, we were interested in the role of circABCB10 in the modulation of LSCC progression. Our data demonstrated that the depletion of circABCB10 significantly inhibited the proliferation and induced the apoptosis of LSCC cells. Meanwhile, circABCB10 knockdown was able to remarkably reduce the invasion and migration of LSCC cells. Mechanically, circABCB10 served as a sponge for microRNAs-588 (miR-588) and miR-588 could target and down-regulated chemokine receptor 4 (CXCR4) expression in LSCC cells. The overexpression of CXCR4 or miR-588 inhibitor could reverse circABCB10 depletion-attenuated malignant phenotypes of LSCC cells. Functionally, the depletion of circABCB10 alleviated the tumor growth of LSCC cells in the tumorigenicity analysis of nude mice. The CXCR4 expression was decreased while the miR-588 expression was enhanced by circABCB10 depletion in vivo. Thus, we concluded that circABCB10 was involved in the malignant progression of LSCC by regulating miR-588/CXCR4 axis. Our finding provides new insights into the mechanism of circRHOT1 contributing to the development of LSCC. CircABCB10 and miR-588 may be used as potential targets for the treatment of LSCC.

Mitochondria-Targeted BODIPY Nanoparticles for Enhanced Photothermal and Photoacoustic Imaging In Vivo.

Short-wavelength absorption and emission (<600 nm), hydrophobicity, and low selectivity have greatly restricted the biomedical applications of BODIPY. Herein, a series of mitochondria-targeted BODIPY nanoparticles with a cationic triphenylphosphine (TPP) group (Mito-BDP1-5 NPs) bearing different lengths of ethylene glycol (0-4 units), along with HO-BDP5 without a cationic TPP group for comparison, have been rationally designed and prepared to investigate the interplay between their structures and the related properties. Our studies found that Mito-BDP1-4 NPs showed a tendency of aggregation and precipitation while Mito-BDP5 NPs could be stable in aqueous solutions. Compared with HO-BDP5, tailor-made Mito-BDP5 possessed a high photothermal conversion efficiency (PCE) of 76.6 vs 9.0% and exhibited the highest photoinduced cytotoxicity. Upon NIR irradiation, the temperature induced by Mito-BDP5 NPs increased rapidly from room temperature to 76.0 °C in vitro and 50.0 °C at the tumor site in vivo within 5 min. Furthermore, effective mitochondrial imaging in vitro, photothermal imaging (PTI), and photoacoustic imaging (PAI) in vivo were achieved. In this paper, we developed tailor-made photothermal agents for targeting mitochondria and enhancing the PTI and PAI performances, which could be potentially applied in clinical precision theranostics.

Anomalous Proton Transport across Silica Nanochannel Membranes Investigated by Ion Conductance Measurements.

Proton transport plays an important role in many biological and technological processes. Numerous experiments and molecular dynamics simulations have proved the increase of proton mobility in confined nanostructures. In this work, we studied the proton transport across flow-through silica nanochannel membranes (SNMs) with vertically aligned channels, uniform diameter (∼2.3 nm), high porosity (16.7%), and ultrasmall thickness (88 nm). Taking into account both the mutual interaction between nanochannels and the contribution of surface conductance, a new theoretical model of ion conductance for SNMs was derived by modifying the conductance model reported previously with a correction factor. The correction factor was estimated by closely matching the experimental conductance of SNMs in KCl and NaCl solutions with the theoretical one calculated by the model. Then the measured conductance of SNMs in HCl solutions was found to be at least four times higher than the calculated value by the model. Given the total conductance across SNMs is dominated by the access conductance instead of channel conductance, the difference between experimental and theoretical conductance values implies either that the theoretical model does not capture the real physics of access conductance or that the two-dimensional nanoconfinement effect exists at the nanochannel entrances. The latter effect likely arises from mutual interaction of neighboring nanochannel entrances.

Highly Efficient Desalting by Silica Isoporous Membrane-Based Microfluidic Chip for Electrospray Ionization Mass Spectrometry.

Nonvolatile buffers and inorganic salts used for isolation and stabilization of biological samples are essential to be cleaned up prior to mass spectrometry (MS) analysis because of their deleterious effects such as ion suppression and instrumental pollution. In this work, a centimeter-scale continuous silica isoporous membrane (SIM) was prepared and integrated into a facile microfluidic chip for the desalting of protein samples based on dialysis principle. Thanks to the uniform pore size (∼2.3 nm in diameter), ultrasmall thickness (90 nm) and high pore density (4.0 × 1012 pores cm-2, corresponding to a porosity of 16.7%) of SIM, the device achieved ∼99% desalting efficiency for the sample with 154 mM NaCl (isotonic saline) at a flow rate of 1 µL min-1, while protein loss was only 5%. High-quality electrospray ionization (ESI)-MS spectra of cytochrome c dissolved in isotonic saline was obtained after the desalting treatment. In addition, the SIM-based microfluidic device was successfully online-coupled with microchip ESI-MS for real-time desalting and characterization of proteins.

pH-Controlled Drug Release by Diffusion through Silica Nanochannel Membranes.

We report in this work the fabrication of a flow-through silica nanochannel membrane (SNM) for controlled drug release applications. The ultrathin SNM consists of parallel nanochannels with a uniform diameter of ∼2.3 nm and a density of 4 × 1012 cm-2, which provide simultaneously high permeability and size selectivity toward small molecules. The track-etched porous polyethylene terephthalate film premodified with silane on its surface was used to support the ultrathin SNM via irreversible covalent bond formation, thus offering mechanical strength, flexibility, and stability to the ultrathin SNM for continuous and long-term use. Alkylamines were subsequently grafted onto the SNM surface to modulate the "on" and "off" state of nanochannels by medium pH for controlled drug release. Thiamphenicol glycinate hydrochloride (TPG), an intestinal drug, was studied as a model to permeate through an ultrathin SNM in both simulated gastric fluid (pH = 1.2) and simulated intestinal fluid (pH = 7.5). The release in the latter case was 178 times faster than that in the former. Moreover, a nearly zero-order constant release of TPG via single-file diffusion was achieved up to 24 h, demonstrating the feasibility of sustained and continuous release of small-molecule drugs in a pH-controlled manner.

Silica-Nanochannel-Based Interferometric Sensor for Selective Detection of Polar and Aromatic Volatile Organic Compounds.

Public awareness of the toxicity of volatile organic compounds (VOCs) has led to increased requirements for direct measurement of these substances. This work reports the sensitive detection of VOCs at the ppb level by an interferometric sensor based on the multilayer silica-nanochannel membrane (MSNM). The MSNM is fabricated by layer-by-layer stacking of a free-standing ultrathin SNM composed of regularly ordered channels with an ultrasmall diameter of about 2.3 nm and an ultrahigh density of about 4 × 1012 cm-2. Light reflected from parallel interfaces of the MSNM gives rise to the interferometric pattern with constructive and destructive fringes. The adsorption of VOCs to a highly porous MSNM varies the refractive index of the MSNM, resulting in the shift of the reflectometric interference spectrum (RIS) and thus yielding highly sensitive responses with a limit of detection (LOD) at the ppb level. Moreover, the sensor selectively responds to polar ethanol and acetone, as well as aromatic benzene, toluene, and chlorobenzene, but is insensitive to nonpolar ethane or hexane. The selectivity most likely arises from hydrogen bonding and dipole interaction of VOCs with silica surface.

Numerical Simulation and Clinical Verification of the Minimally Invasive Repair of Pectus Excavatum.

OBJECTIVE: In this article we proposed a modeling method by building an assembled model to simulate the orthopedic process of minimally invasive surgery for pectus excavatum and got the clinical verification, which aims to provide some references for clinic diagnoses, treatment, and surgery planning. METHODS: The anterior chest model of a 15-year-old patient was built based on his CT images; and his finite element model and the Nuss bar were created. Coupling of nodal displacement was used to connect bones with cartilages of the anterior chest. Turning the Nuss bar over is completed by rotating displacement of it. By comparing the numerical simulation outcomes with clinical surgery results, the numerical simulation results were verified. RESULTS: The orthopedic process of minimally invasive surgery of pectus excavatum was simulated by model construction and numerical analysis. The stress, displacement fields and distribution of the contact pressure between the Nuss bar and costal cartilages were analyzed. The relationship between correcting force and displacement was obtained. Compared with the of clinical results, the numerical simulation results were close to that of the actual clinical surgery in displacement field, and the final contact position of the Nuss bar and the costal cartilages. CONCLUSION: Compared with the rigid model, the assembled simulation model is in more conformity with the actual clinical practice. The larger curvature results in the maximum equivalent stress, which is the main reason for clinical pain. Soft tissues and muscles should be taken into account in the numerical simulation process.