Empagliflozin ameliorates vascular calcification in diabetic mice through inhibiting Bhlhe40-dependent NLRP3 inflammasome activation.

Type 2 diabetes mellitus (T2DM) patients exhibit greater susceptibility to vascular calcification (VC), which has a higher risk of death and disability. However, there is no specific drug for VC therapy. NLRP3 inflammasome activation as a hallmark event of medial calcification leads to arterial stiffness, causing vasoconstrictive dysfunction in T2DM. Empagliflozin (EMPA), a sodium-glucose co-transporter 2 inhibitor (SGLT2i), restrains hyperglycemia with definite cardiovascular benefits. Given the anti-inflammatory activity of EMPA, herein we investigated whether EMPA protected against VC in the aorta of T2DM mice by inhibiting NLRP3 inflammasome activation. Since db/db mice receiving a normal diet developed VC at the age of about 20 weeks, we administered EMPA (5, 10, 20 mg·kg-1·d-1, i.g) to 8 week-old db/db mice for 12 weeks. We showed that EMPA intervention dose-dependently ameliorated the calcium deposition, accompanied by reduced expression of RUNX2 and BMP2 proteins in the aortas. We found that EMPA (10 mg·kg-1·d-1 for 6 weeks) also protected against VC in vitamin D3-overloaded mice, suggesting the protective effects independent of metabolism. We showed that EMPA (10 mg·kg-1·d-1) inhibited the abnormal activation of NLRP3 inflammasome in aortic smooth muscle layer of db/db mice. Knockout (KO) of NLRP3 significantly alleviated VC in STZ-induced diabetic mice. The protective effects of EMPA were verified in high glucose (HG)-treated mouse aortic smooth muscle cells (MOVASs). In HG-treated NLRP3 KO MOVASs, EMPA (1 µM) did not cause further improvement. Bioinformatics and Western blot analysis revealed that EMPA significantly increased the expression levels of basic helix-loop-helix family transcription factor e40 (Bhlhe40) in HG-treated MOVASs, which served as a negative transcription factor directly binding to the promotor of Nlrp3. We conclude that EMPA ameliorates VC by inhibiting Bhlhe40-dpendent NLRP3 inflammasome activation. These results might provide potential significance for EMPA in VC therapy of T2DM patients.

Lonicera japonica protects Pelodiscus sinensis by inhibiting the biofilm formation of Aeromonas hydrophila.

Aquaculture has suffered significant financial losses as a result of the infection of zoonotic Aeromonas hydrophila, which has a high level of resistance to classic antibiotics. In this study, we isolated an A. hydrophila strain B3 from diseased soft-shelled turtle (Pelodiscus sinensis), which is one of the most commercially significant freshwater farmed reptiles in East Asia, and found that A. hydrophila was its dominant pathogen. To better understand the inhibition effect and action mechanism of Chinese herbs on A. hydrophila, we conducted Chinese herbs screening and found that Lonicera japonica had a significant antibacterial effect on A. hydrophila B3. Experimental therapeutics of L. japonica on soft-shelled turtle showed that the supplement of 1% L. japonica to diet could significantly upregulate the immunity-related gene expression of soft-shelled turtle and protect soft-shelled turtle against A. hydrophila infection. Histopathological section results validated the protective effect of L. japonica. As the major effective component of L. japonica, chlorogenic acid demonstrated significant inhibitory effect on the growth of A. hydrophila with MIC at 6.4 mg/mL. The in vitro assay suggested that chlorogenic acid could inhibit the hemolysin/protease production and biofilm formation of A. hydrophila and significantly decrease the expression of quorum sensing, biofilm formation, and hemolysin-related genes in A. hydrophila. Our results showed that the Chinese herb L. japonica would be a promising candidate for the treatment of A. hydrophila infections in aquaculture, and it not only improves the immune response of aquatic animals but also inhibits the virulence factor (such as biofilm formation) expression of A. hydrophila. KEY POINTS: • A. hydrophila was the dominant pathogen of the diseased soft-shelled turtle. • L. japonica can protect soft-shelled turtle against A. hydrophila infection. • Chlorogenic acid inhibits the growth and biofilm formation of A. hydrophila.

Three chemosensory proteins enriched in antennae and tarsi of Rhaphuma horsfieldi differentially contribute to the binding of insecticides.

Antennae and legs (primarily the tarsal segments) of insects are the foremost sensory organs that contact a diverse range of toxic chemicals including insecticides. Binding proteins expressed in the two tissues are potential molecular candidates serving as the binding and sequestering of insecticides, like chemosensory proteins (CSPs). Insect CSPs endowed with multiple roles have been suggested to participate in insecticide resistance, focusing mainly on moths, aphids and mosquitos. Yet, the molecular underpinnings underlying the interactions of cerambycid CSPs and insecticides remain unexplored. Here, we present binding properties of three antenna- and tarsus-enriched RhorCSPs (RhorCSP1, CSP2 and CSP3) in Rhaphuma horsfieldi to eight insecticide classes totaling 15 chemicals. From the transcriptome of this beetle, totally 16 CSP-coding genes were found, with seven full-length sequences. In phylogeny, these RhorCSPs were distributed dispersedly in different clades. Expression profiles revealed the abundant expression of RhorCSP1, CSP2 and CSP3 in antennae and tarsi, thus as representatives for studying the protein-insecticide interactions. Binding assays showed that the three RhorCSPs were tuned differentially to insecticides but exhibited the highest affinities with hexaflumuron, chlorpyrifos and rotenone (dissociation constants <13 µM). In particular, RhorCSP3 could interact strongly with 10 of tested insecticides, of which four residues (Tyr25, Phe42, Val65 and Phe68) contributed significantly to the binding of six, four, three and four ligands, respectively. Of these, the binding of four mutated RhorCSP3s to a botanical insecticide rotenone was significantly weakened compared to the wildtype protein. Furthermore, we also evidenced that RhorCSP3 was a broadly-tuned carrier protein in response to a wide variety of plant odorants outside insecticides. Altogether, our findings shed light on different binding mechanisms and odorant-tuning profiles of three RhorCSPs in R. horsfieldi and identify key residues of the RhorCSP3-insecticide interactions.

CRISPR/Cas9-mediated ebony knockout causes melanin pigmentation and prevents moth Eclosion in Ectropis grisescens.

Ectropis grisescens (Lepidoptera: Geometridae) is a destructive tea pest in China. Mimesis, characterized by changing body color, is an important trait of E. grisescens larvae. Hence, identifying melanin pathway-related genes may contribute to developing new pest control strategies. In the present study, we cloned Egebony, a gene potentially involved in melanin pigmentation in E. grisescens, and subsequently conducted CRISPR/Cas9-mediated targeted mutagenesis of Egebony to analyze its role in pigmentation and development. At the larvae, prepupae, and pupae stages, Egebony-knockout individuals exhibited darker pigmentation than the wild-type. However, Egebony knockout did not impact the colors of sclerotized appendants, including ocelli, setae, and claws. While mutant pupae could successfully develop into moths, they were unable to emerge from the puparium. Notably, embryo hatchability and larval survival of mutants remained normal. Further investigation indicated that mutant pupae exhibited significantly stronger shearing force than the wild-type, with the pigmented layer of mutant pupae appearing darker and thicker. Collectively, these results suggest that the loss of Egebony might increase the rigidity of the puparium and prevent moth eclosion. This study provides new insights into understanding the function and diversification of ebony in insect development and identifies a lethal gene that can be manipulated for developing effective pest control strategies.

Neural network analysis of pharyngeal sounds can detect obstructive upper respiratory disease in brachycephalic dogs.

Brachycephalic obstructive airway syndrome (BOAS) is a highly prevalent respiratory disease affecting popular short-faced dog breeds such as Pugs and French bulldogs. BOAS causes significant morbidity, leading to poor exercise tolerance, sleep disorders and a shortened lifespan. Despite its severity, the disease is commonly missed by owners or disregarded by veterinary practitioners. A key clinical sign of BOAS is stertor, a low-frequency snoring sound. In recent years, a functional grading scheme has been introduced to semi-objectively grade BOAS based on the presence of stertor and other abnormal signs. However, correctly grading stertor requires significant experience and adding an objective component would aid accuracy and repeatability. This study proposes a recurrent neural network model to automatically detect and grade stertor in laryngeal electronic stethoscope recordings. The model is developed using a novel dataset of 665 labelled recordings taken from 341 dogs with diverse BOAS clinical signs. Evaluated via nested cross validation, the neural network predicts the presence of clinically significant BOAS with an area under the receiving operating characteristic of 0.85, an operating sensitivity of 71% and a specificity of 86%. The algorithm could enable widespread screening for BOAS to be conducted by both owners and veterinarians, improving treatment and breeding decisions.

Candidate membrane protein gene families related to chemoreception in a wood-boring beetle, Pharsalia antennata Gahan (Coleoptera: Cerambycidae).

The longhorned beetles are key players for the maintenance of biodiversity in the terrestrial ecosystem. As xylophagous cerambycid insects in Coleoptera, the beetles have evolved specialized olfactory and gustatory systems to recognize chemical cues in the surrounding habitats. Despite over 36,000 described species in the Cerambycidae family including a wood-boring pest Pharsalia antennata, only a limited number of them (<1 %) have been characterized regarding their chemical ecology at the molecular level. Here, we surveyed four membrane protein gene families in P. antennata related to chemoreception through transcriptomics, phylogenetics and expression profiling analyses. In total, 144 genes encoding 72 odorant receptors (ORs), 33 gustatory receptors (GRs), 23 ionotropic receptors (IRs), four sensory neuron membrane proteins (SNMPs) and 12 ionotropic glutamate receptors (iGluRs) were harvested from the transcriptome of multiple tissues including antennae and legs of both sexes. The lineage-specific expansion of PantORs possibly implied a diverse range of host plants in this beetle, supporting this correlation between the host range and olfactory receptor repertoire sizes across cerambycid species. Further phylogenetic analysis revealed that Group 2 was contributed mainly to the large OR gene repertoire in P. antennata, representing 18 genes in Group 2A and eight in Group 2B. On the other hand, some key chemosensory genes were identified by applying a phylogenetics approach, such as PantOR21 close to the 2-phenylethanol receptor in Megacyllene caryae, three carbon dioxide GRs and seven Antennal IRs (A-IRs) clades. We also determined sex- and tissue-specific expression profiles of 69 chemosensory genes, revealing the high expression of most PantORs in antennae. Noticeably, 10 sex-biased genes (six PantORs, three PantIRs and PantSNMP1a) were presented in antennae, five sex-biased PantGRs in legs and 39 sex-biased genes (15 PantORs, 13 PantGRs, eight PantIRs and three PantSNMPs) in abdomens. These findings have greatly enhanced our knowledge about the chemical ecology of P. antennata and identify candidate molecular targets for mediating smell and taste of this beetle.

The supplementation of Rothia as a potential preventive approach for bone loss in mice with ovariectomy-induced osteoporosis.

There is an inseparable link between bone metabolism and gut microbiota, and the supplementation of probiotics exhibits a significant role in maintaining the homeostasis of gut microbiota and inhibiting bone loss. This study aims to explore the preventive and therapeutic potentials and the specific mechanisms of Rothia on osteoporosis. The mice models of osteoporosis induced by ovariectomy (OVX) were built, and the regular (once a day) and quantitative (200 µL/d) gavage of Rothia was performed for 8 weeks starting from 1 week after OVX. Microcomputed tomography was used to analyze the bone mass and bone microstructure of mice in each group after sacrifice. Histological staining and immunohistochemistry were then applied to identify the expression of pro-inflammatory cytokines, intestinal permeability, and osteogenic and osteoclastic activities of mice. The collected feces of mice in each group were used for 16S rRNA high-throughput sequencing to detect the alterations in composition, abundance, and diversity of gut microbiota. This study demonstrated that the gavage of Rothia alleviated bone loss in mice with OVX-induced osteoporosis, improved OVX-induced intestinal mucosal barrier injury, optimized intestinal permeability (zonula occludens protein 1 and occludin), reduced intestinal inflammation (tumor necrosis factor-α and interleukin-1ß), and regulated imbalance of gut microbiota. Based on "gut-bone" axis, this study revealed that regular and quantitative gavage of Rothia can relieve bone loss in mice with OVX-induced osteoporosis by repairing the intestinal mucosal barrier injury, optimizing the intestinal permeability, inhibiting the release of pro-inflammatory cytokines, and improving the disorder of gut microbiota.

The Clinical Impact of Different Types of Preoperative Biliary Intervention on Postoperative Biliary Tract Infection of Patients Undergoing Pancreaticoduodenectomy.

Background: For patients with obstructive jaundice and who are indicated for pancreaticoduodenectomy (PD) or biliary intervention, either endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic cholangiography and drainage (PTCD) may be indicated preoperatively. However, the possibility of procedure-related postoperative biliary tract infection (BTI) should be a concern. We tried to evaluate the impact of ERCP and PTCD on postoperative BTI. Methods: Patients diagnosed from June 2013 to March 2022 with periampullary lesions and with PD indicated were enrolled in this cohort. Patients without intraoperative bile culture and non-neoplastic lesions were excluded. Clinical information, including demographic and laboratory data, pathologic diagnosis, results of microbiologic tests, and relevant infectious outcomes, was extracted from medical records for analysis. Results: One-hundred-and-sixty-four patients from the cohort (164/689) underwent preoperative biliary intervention, either ERCP (n = 125) or PTCD (n = 39). The positive yield of intraoperative biliary culture was significantly higher in patients who underwent ERCP than in PTCD (90.4% vs. 41.0%, p < 0.001). Although there was no significance, a trend of higher postoperative BTI (13.8% vs. 2.7%) and BTI-related septic shock (5 vs. 0, 4.0% vs. 0%) in the ERCP group was noticed. While the risk factors for postoperative BTI have not been confirmed, a trend suggesting a higher incidence of BTI associated with ERCP procedures was observed, with a borderline p-value (p = 0.05, regarding ERCP biopsy). Conclusions: ERCP in patients undergoing PD increases the positive yield of intraoperative biliary culture. PTCD may be the favorable option if preoperative biliary intervention is indicated.

Computed tomography medical image analysis of basketball players' bone geometry / Análise de imagens médicas por tomografia computadorizada da geometria óssea de jogadores de basquete / Análisis de imágenes médicas por tomografía computarizada de la geometría ósea de jugadores de baloncesto

ABSTRACT Introduction: Basketball sports will effect on the morphology and structure of the hand bones and joints. Objective: The article selected nine professional basketball players (basketball group) and 11 non-basketball players (control group) in the provincial youth team. A 64-row spiral computed tomography scan was used to scan the wrist and three-dimensional reconstruction. The volume of each carpal bone and the computed tomography value (bone density) were measured. Methods: To explore the influence of basketball sports on the hand bones and joints' morphological structure, the paper analyzes the structural characteristics of the computed tomography images of young male basketball players' wrist bones. Results: Compared with the carpal bones in the control group, the volume of the right navicular bone and the small polygonal bone, the left-hand navicular bone, the large triangular bone, and the small polygonal bone in the basketball group increased significantly (P<0.05). Conclusions: Basketball can increase the volume of the part of the wrist bones of adolescent male athletes and reduce the bone density; the morphological structure of the non-smashing wrist bones of basketball players has similar changes to that of the spikers. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: O basquetebol afetará a morfologia e a estrutura dos ossos e articulações da mão. Objetivo: o artigo selecionou nove jogadores profissionais de basquete (grupo de basquete) e 11 não jogadores de basquete (grupo de controle) da equipe juvenil da província. Uma tomografia computadorizada espiral de 64 linhas foi usada para digitalizar o punho e a reconstrução tridimensional. O volume de cada osso do carpo e o valor da tomografia computadorizada (densidade óssea) foram medidos. Métodos: Para explorar a influência dos esportes de basquete sobre os ossos da mão e a estrutura morfológica das articulações, o artigo analisa as características estruturais das imagens de tomografia computadorizada de ossos do punho de jovens jogadores de basquete do sexo masculino. Resultados: Em comparação com os ossos do carpo no grupo de controle, o volume do osso navicular direito e o osso poligonal pequeno, o osso navicular esquerdo, o osso triangular grande e o osso poligonal pequeno no grupo de basquete aumentaram significativamente (P <0,05). Conclusões: O basquete pode aumentar o volume dos ossos do punho de atletas adolescentes do sexo masculino e reduzir a densidade óssea; a estrutura morfológica dos ossos do pulso que não se quebram em jogadores de basquete tem mudanças semelhantes às dos espigões. Nível de evidência II; Estudos terapêuticos- investigação dos resultados do tratamento.

RESUMEN Introducción: el baloncesto afectará la morfología y la estructura de los huesos y articulaciones de la mano. Objetivo: El artículo seleccionó nueve jugadores de baloncesto profesionales (grupo de baloncesto) y 11 jugadores no baloncesto (grupo de control) en el equipo juvenil provincial. Se utilizó una tomografía computarizada en espiral de 64 filas para escanear la muñeca y la reconstrucción tridimensional. Se midieron el volumen de cada hueso del carpo y el valor de la tomografía computarizada (densidad ósea). Métodos: Para explorar la influencia de los deportes de baloncesto en la estructura morfológica de los huesos de la mano y las articulaciones, el artículo analiza las características estructurales de las imágenes de tomografía computarizada de los huesos de la muñeca de los jóvenes jugadores de baloncesto. Resultados: En comparación con los huesos del carpo en el grupo de control, el volumen del hueso navicular derecho y el hueso poligonal pequeño, el hueso navicular izquierdo, el hueso triangular grande y el hueso poligonal pequeño en el grupo de baloncesto aumentaron significativamente (P <0,05). Conclusiones: El baloncesto puede aumentar el volumen de los huesos de la muñeca de los deportistas varones adolescentes y reducir la densidad ósea; la estructura morfológica de los huesos de las muñecas que no se rompen de los jugadores de baloncesto tiene cambios similares a los de los atacantes. Nivel de evidencia II; Estudios terapéuticos- investigación de los resultados del tratamiento.

Research on knee sports injury classification based on medical images and kwon3d software / Pesquisa sobre classificação de lesões esportivas no joelho com base em imagens médicas e software kwon3d / Investigación sobre clasificación de lesiones deportivas de rodilla basada en imágenes médicas y software kwon3d

ABSTRACT Introduction: The integrity of articular cartilage determines the functional state of the joint. In recent years, the development of MRI sequences of various articular cartilage has become the focus of many research topics. Objective: The accuracy of diagnosis of knee cartilage injury caused by motion injury was studied retrospectively by meta-three-dimensional software. Methods: Forty-six knee joints of 45 patients with sports injuries, multi-sequence MRI was performed before surgery, including conventional knee MRI (SET1WI, FSEPD/T2WI), 3D SPGR, and 3D FIESTA sequences. Results: According to the operation results, the sensitivity, specificity, positive predictive value, and negative predictive value of 3D SPGR combined with conventional MRI sequence evaluation of cartilage damage are the highest, 73%, 98%, 95%, and 90%. Conclusions: 3D SPGR combined with conventional MRI sequences can improve accurate evaluation and diagnosis of cartilage disease over a reasonable scan time. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: A integridade da cartilagem articular determina o estado funcional da articulação. Nos últimos anos, o desenvolvimento de sequências de ressonância magnética de várias cartilagens articulares se tornou o foco de muitos tópicos de pesquisa. Objetivo: A precisão do diagnóstico de lesão da cartilagem do joelho causada por lesão de movimento foi estudada retrospectivamente por software meta-tridimensional. Métodos: Quarenta e seis articulações de joelho de 45 pacientes com lesões esportivas, várias sequências de ressonância magnética foram realizadas antes da cirurgia, incluindo ressonância magnética de joelho convencional (SET1WI, FSEPD / T2WI), 3D SPGR e sequências 3D FIESTA. Resultados: De acordo com os resultados da operação, a sensibilidade, especificidade, valor preditivo positivo e valor preditivo negativo de 3D SPGR combinado com avaliação de sequência de ressonância magnética convencional de danos na cartilagem são os mais altos, 73%, 98%, 95% e 90%. Conclusões: 3D SPGR combinado com sequências convencionais de ressonância magnética pode melhorar a avaliação precisa e diagnóstico de doença da cartilagem em um tempo de varredura razoável. Nível de evidência II; Estudos terapêuticos- investigação dos resultados do tratamento.

RESUMEN Introducción: La integridad del cartílago articular determina el estado funcional de la articulación. En los últimos años, el desarrollo de secuencias de resonancia magnética de varios cartílagos articulares se ha convertido en el foco de muchos temas de investigación. Objetivo: La precisión del diagnóstico de la lesión del cartílago de la rodilla causada por una lesión por movimiento se estudió retrospectivamente mediante un software meta-tridimensional. Métodos: Cuarenta y seis articulaciones de rodilla de 45 pacientes con lesiones deportivas, se realizó una resonancia magnética de secuencia múltiple antes de la cirugía, incluida la resonancia magnética de rodilla convencional (SET1WI, FSEPD/T2WI), secuencias 3D SPGR y 3D FIESTA. Resultados: De acuerdo con los resultados de la operación, la sensibilidad, la especificidad, el valor predictivo positivo y el valor predictivo negativo de 3D SPGR combinados con la evaluación de la secuencia de resonancia magnética convencional del daño del cartílago son los más altos, 73%, 98%, 95% y 90%. Conclusiones: 3D SPGR combinado con secuencias de resonancia magnética convencionales puede mejorar la evaluación y el diagnóstico precisos de la enfermedad del cartílago en un tiempo de exploración razonable. Nivel de evidencia II; Estudios terapéuticos- investigación de los resultados del tratamiento.

Enhancement of motor functional recovery in thoracic spinal cord injury: voluntary wheel running versus forced treadmill exercise.

JOURNAL/nrgr/04.03/01300535-202503000-00028/figure1/v/2024-06-17T092413Z/r/image-tiff Spinal cord injury necessitates effective rehabilitation strategies, with exercise therapies showing promise in promoting recovery. This study investigated the impact of rehabilitation exercise on functional recovery and morphological changes following thoracic contusive spinal cord injury. After a 7-day recovery period after spinal cord injury, mice were assigned to either a trained group (10 weeks of voluntary running wheel or forced treadmill exercise) or an untrained group. Bi-weekly assessments revealed that the exercise-trained group, particularly the voluntary wheel exercise subgroup, displayed significantly improved locomotor recovery, more plasticity of dopaminergic and serotonin modulation compared with the untrained group. Additionally, exercise interventions led to gait pattern restoration and enhanced transcranial magnetic motor-evoked potentials. Despite consistent injury areas across groups, exercise training promoted terminal innervation of descending axons. In summary, voluntary wheel exercise shows promise for enhancing outcomes after thoracic contusive spinal cord injury, emphasizing the role of exercise modality in promoting recovery and morphological changes in spinal cord injuries. Our findings will influence future strategies for rehabilitation exercises, restoring functional movement after spinal cord injury.

Chondroitinase ABC combined with Schwann cell transplantation enhances restoration of neural connection and functional recovery following acute and chronic spinal cord injury.

JOURNAL/nrgr/04.03/01300535-202505000-00029/figure1/v/2024-07-28T173839Z/r/image-tiff Schwann cell transplantation is considered one of the most promising cell-based therapy to repair injured spinal cord due to its unique growth-promoting and myelin-forming properties. A the Food and Drug Administration-approved Phase I clinical trial has been conducted to evaluate the safety of transplanted human autologous Schwann cells to treat patients with spinal cord injury. A major challenge for Schwann cell transplantation is that grafted Schwann cells are confined within the lesion cavity, and they do not migrate into the host environment due to the inhibitory barrier formed by injury-induced glial scar, thus limiting axonal reentry into the host spinal cord. Here we introduce a combinatorial strategy by suppressing the inhibitory extracellular environment with injection of lentivirus-mediated transfection of chondroitinase ABC gene at the rostral and caudal borders of the lesion site and simultaneously leveraging the repair capacity of transplanted Schwann cells in adult rats following a mid-thoracic contusive spinal cord injury. We report that when the glial scar was degraded by chondroitinase ABC at the rostral and caudal lesion borders, Schwann cells migrated for considerable distances in both rostral and caudal directions. Such Schwann cell migration led to enhanced axonal regrowth, including the serotonergic and dopaminergic axons originating from supraspinal regions, and promoted recovery of locomotor and urinary bladder functions. Importantly, the Schwann cell survival and axonal regrowth persisted up to 6 months after the injury, even when treatment was delayed for 3 months to mimic chronic spinal cord injury. These findings collectively show promising evidence for a combinatorial strategy with chondroitinase ABC and Schwann cells in promoting remodeling and recovery of function following spinal cord injury.