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Purpose: To evaluate the response of human peri-implant soft tissue (PIST) to different healing abutment materials 24 hours after positioning by assessing the expression of genes related to the early connective tissue wound healing response. Materials and Methods: The following four materials were used to create experimental abutments that were mounted on implants placed in five patients (four different abutments in each patient): group A--grade 4 titanium (Ti), group B--grade 5 Ti, group C--zirconia (Zr), and group D--polyetheretherketone (PEEK). Before implant placement, a gingival biopsy (control, CT) sample was obtained using a 2-mm-diameter punch (T0). After 24 hours, PIST biopsy samples were collected using a specifically designed custom-made cutting device. Real-time polymerase chain reaction (PCR) was performed to analyze the expression of the following genes: COL-I, COL-III, MMP-1, TIMP-1, TGF-^1, FN, ITGA4, ITGA5, ITGB1, RAC-1, COL-IV, aSMA, IL-6, and CXCL-1. Results: Gene expression analysis showed some differences between CT and the experimental groups; however, no significant differences were detected when comparing the experimental groups. COL-I was significantly downregulated in groups A and C compared to CT. Expression of MMP-1 and TIMP-1 increased in all the experimental groups but to a lesser extent in group A. FN, RAC-1, COL-IV, and aSMA were downregulated, especially in group A, in which CXCL-1 and IL-6 showed the lowest expression. Conclusions:The results of grade 4 Ti and Zr abutments seem to be promising, because a lower expression of genes related to inflammation, myofibroblast activation, and extracellular matrix remodeling was observed when compared with grade 5 Ti and PEEK, without triggering a profibrotic response in the early phases of PIST repair. [ABSTRACT FROM AUTHOR]

Purpose: To examine the behavior and function of human gingival fibroblasts growing on healing abutments with or without laser-textured topography. Materials and Methods: Human primary gingival connective tissue fibroblasts were cultured on healing abutments with machined or laser-textured (Laser-Lok, BioHorizons) surfaces. Cellular and molecular responses were evaluated by a variety of tests, including cell density assay (WST-1), fluorescence microscopy, real-time quantitative reverse-transcription polymerase chain reaction (qRT-PCR), and detachment tests. Results: The machined surface showed monodirectional traces and scratches from milling, whereas the laser-textured surface showed a distinct morphology consisting of monodirectional mesoscale channels (15-µm pitch) and woven oblique microridges formed within the channels. There were no differences in initial fibroblast attachment, subsequent fibroblast proliferation, or collagen production between the machined and laser-textured surfaces. Fibroblasts growing on a laser-textured surface were found to spread in one direction along the mesochannels, while cells growing on machined surfaces tended to spread randomly. Fibroblasts on laser-textured surfaces were 1.8 times more resistant to detachment than those on machined surfaces. An adhesive glycoprotein (fibronectin) and transmembrane adhesion linker gene (integrin ß-1) were upregulated on laser-textured surfaces. Conclusions: The increased fibroblast retention, uniform growth, and increased transcription of cell adhesion proteins compellingly explain the enhanced tissue-level response to laser-created and hybrid-textured titanium surfaces. These results provide a cellular and molecular rationale for the tissue reaction to this unique surface; in addition, they support its extended use, from implants and healing abutments to diverse prosthetic components where enhanced soft tissue responses would be desirable. [ABSTRACT FROM AUTHOR]

The grapevine fleck virus (GFkV) is a ubiquitous grapevine-infecting virus found worldwide, is associated with the grapevine fleck complex, and is often found in mixed infections with viruses of the grapevine leafroll complex and/or vitiviruses. Although GFkV has been studied for a long time, limited sequence information is available in the public databases. In this study, the GFkV sequence data available in GenBank and data generated at the Foundation Plant Services, University of California, Davis, were used to perform nucleotide sequence comparisons, construct a phylogenetic tree, and develop a new RT-qPCR assay. Sequence comparisons showed high genetic diversity among the GFkV isolates, and the phylogenetic analyses revealed a new group comprised of GFkV isolates identified in the present study. A new assay, referred to as GFkV-CP, was designed and validated using an existing GFkV positive control together with 11 samples known to be infected with combinations of different marafiviruses and maculaviruses but not GFkV. In addition, the newly designed assay was used in a field survey to screen grapevines from diverse geographical locations that are maintained at the United States Department of Agriculture (USDA) National Clonal Germplasm Repository (NCGR) in Winters, CA.

Esophageal cancer is a grave malignant condition. While radiotherapy, often in conjunction with chemotherapy, serves as a cornerstone in the management of locally advanced or metastatic cases, patient tolerance and treatment resistance frequently hinder its efficacy. Cell-in-cell structures, prevalent in various tumors, have been linked to prognosis. Hence, investigating the prognostic significance and regulatory mechanisms of genes related to these intracellular structures in esophageal cancer is imperative. The Cancer Genome Atlas (TCGA) Esophageal Cancer (ESCA) dataset served as the training set for the analysis. Differentially expressed genes (DEGs) in ESCA samples were identified, with those related to intercellular structures designated cell-in-cell-related differential expression genes (CIC-related DEGs). Cox regression analysis was employed to identify prognostic genes, categorizing samples into high- and low-risk groups based on median risk scores. Validation was conducted using the GSE53624 risk model. Established methodologies included morphological mapping, enrichment analysis, immune infiltration analysis, prognostic gene expression validation, molecular docking, and Reverse Transcription Polymerase Chain Reaction (RT-PCR) validation. Thirty-eight intersecting genes were identified between the disease and normal groups in ESCA samples. Stepwise multivariate Cox analysis pinpointed three prognostic genes: androgen receptor (AR), C-X-C motif chemokine ligand 8 (CXCL8), and epidermal growth factor receptor (EGFR). The risk model's applicability was confirmed in the GSE53624 dataset, revealing eight significantly different immune-related gene sets. Prognostic gene expression validation demonstrated significant differences between the disease and normal groups in both datasets. The proteins corresponding to the three prognostic genes interacted with gefitinib and osimertinib. RT-PCR results corroborated the differential expression of prognostic genes in esophageal cancer tissues. This study identified AR, CXCL8, and EGFR as prognostic genes and demonstrated their molecular interactions with gefitinib and osimertinib, providing a foundation for ESCA diagnosis and treatment. [ABSTRACT FROM AUTHOR]

Introduction: Liver fibrosis is a globally prevalent chronic liver disease, often representing the advanced stage of various chronic liver conditions. Despite its widespread occurrence, there is currently no widely accepted or effective treatment for liver fibrosis. However, increasing evidence supports the efficacy of Traditional Chinese Medicine (TCM) in inhibiting the progression of fibrosis. In this study, we explored the effects and potential mechanisms of Qizhu-Ruogan-Granules (QZRG), a formulation from the Affiliated Hospital of the Chengdu University of TCM, on carbon tetrachloride (CCl4)-induced liver fibrosis in mice. Methods: A total of 40 male C57BL/6J mice were randomly divided into five groups (n = 8 per group), with liver fibrosis induced by injecting 10% CCl4 for 15 weeks. From the 7th week onward, QZRG granules were administered orally to the treatment groups at low, medium, and high doses. To assess liver function, serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were measured. Liver morphology and fibrosis were evaluated using hematoxylin-eosin (H&E) and Masson's trichrome staining, while gene and protein expression levels were analyzed through quantitative reverse transcription polymerase chain reaction (RT-PCR) and western blot techniques. Results: The results showed that QZRG granules significantly reduced serum levels of AST, ALT, and ALP in CCl4-treated mice, alleviated liver damage, and reduced collagen accumulation. Furthermore, QZRG granules inhibited the expression of apoptosis-related proteins BAX, Caspase9, Caspase8, and Caspase3, while reducing P2Y14 expression in fibrotic liver tissues. Additionally, QZRG granules suppressed the proliferation of activated hepatic stellate cells. Conclusion: Our findings suggest that QZRG granules may exert anti-fibrotic effects by downregulating P2Y14 expression and effectively slowing the progression of liver fibrosis. [ABSTRACT FROM AUTHOR]

Background: Disturbances in DNA damage repair may lead to cancer. SIRT1, an NAD+-dependent deacetylase, plays a crucial role in maintaining cellular homeostasis through the regulation of processes such as histone posttranslational modifications, DNA repair, and cellular metabolism. However, a comprehensive exploration of SIRT1's involvement in pan-cancer remains lacking. Our study aimed to analyze the role of SIRT1 in pan-cancer to gain a more comprehensive understanding of its role in multiple malignancies. Methods: We systematically examined the role of SIRT1 in pan-cancer by analyzing data from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases. Various tools, including R, Cytoscape, HPA, Archs4, TISIDB, cBioPortal, STRING, GSCALite, and CancerSEA, were used to integrate and analyze SIRT1 gene expression, prognosis, protein interactions, signaling pathways, immune infiltration, and other relevant information. Furthermore, we validated the differential expression of SIRT1 in normal human kidney cells and kidney cancer cell lines via experimental verification. Results: SIRT1 expression was significantly reduced in various cancers and was different across molecular and immune subtypes. SIRT1 is intricately linked to numerous cancer pathways. In most cancer types, increased SIRT1 expression is positively associated with eosinophils, helper T cells, central memory T cells, effector memory T cells, γδ T cells, and Th2 cells. SIRT1 expression is significantly correlated with immune regulatory factors across various cancer types. Quantitative reverse transcription polymerase chain reaction (qRT–PCR) and Western blot (WB) analyses confirmed that SIRT1 is differentially expressed in kidney renal clear cell carcinoma (KIRC). Conclusions: Using an integrative approach involving bioinformatics analysis and experimental validation, we clarified the potential roles and mechanisms of SIRT1 in pan-cancer, providing a theoretical basis for the development of SIRT1-targeted therapies in clinical applications. [ABSTRACT FROM AUTHOR]

Introduction: The deficiency of estrogen correlates with a range of diseases, notably Postmenopausal osteoporosis (PMO) and Parkinson's disease (PD). There is a possibility that PMO and PD may share underlying molecular mechanisms that are pivotal in their development and progression. The objective of this study was to identify critical genes and potential mechanisms associated with PMO by examining co-expressed genes linked to PD. Methods: Initially, pertinent data concerning PMO and PD were obtained from the GWAS database, followed by conducting a Bayesian colocalization analysis. Subsequently, co-expressed genes from the PMO dataset (GSE35956) and the PD dataset (GSE20164) were identified and cross-referenced with estrogen-related genes (ERGs). Differentially expressed genes (DEGs) among PMO, PD, and ERGs were subjected to an array of bioinformatics analyses, including Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analyses, in addition to protein-protein interaction (PPI) network analysis. The study also involved constructing TF-gene interactions, TF-microRNA coregulatory networks, interactions of hub genes with diseases, and validation through quantitative reverse transcription polymerase chain reaction (qRT-PCR). Results: The colocalization analysis uncovered shared genetic variants between PD and osteoporosis, with a posterior probability of colocalization (PPH4) measured at 0.967. Notably, rs3796661 was recognized as a shared genetic variant. A total of 11 genes were classified as DEGs across PMO, PD, and ERGs. Five principal KEGG pathways were identified, which include the p53 signaling pathway, TGF-beta signaling pathway, cell cycle, FoxO signaling pathway, and cellular senescence. Additionally, three hub genes—WT1, CCNB1, and SMAD7—were selected from the PPI network utilizing Cytoscape software. These three hub genes, which possess significant diagnostic value for PMO and PD, were further validated using GEO datasets. Interactions between transcription factors and genes, as well as between microRNAs and hub genes, were established. Ultimately, the expression trends of the identified hub genes were confirmed through qRT-PCR validation. Conclusions: This study is anticipated to offer innovative approaches for identifying potential biomarkers and important therapeutic targets for both PMO and PD. [ABSTRACT FROM AUTHOR]

Background: The laryngo-tracheal framework is a complex structure responsible for several functions, including phonation, airway formation for breathing, and preventing aspiration. These functions rely on precise neurological and structural coordination. The COVID-19 pandemic primarily affected the lungs and upper respiratory tract, and the patients were classified into mild, moderate, and severe categories to determine appropriate management strategies, including home isolation and hospital admission. Mild cases typically underwent home isolation, while moderate-to-severe cases required hospitalization. Moderate cases often necessitated oxygen supplementation through various means such as oxygen masks, non-rebreather masks, and high-flow nasal cannula, while severe cases required mechanical support and emergency procedures like intubation, continuous positive airway pressure, and tracheostomy. These interventions, along with other factors like viral neuritis and comorbid illnesses, result in various laryngo-tracheal sequelae; in this study, we have tried to look for the laryngo-tracheal sequelae in post-COVID-19 patients. Methods: A sample of 348 patients who were diagnosed with COVID-19 using reverse transcription polymerase chain reaction were identified retrospectively, and their demographics, comorbidities, intubation/tracheostomy, and management data were collected; then all the patients were subjected to video-laryngoscopy, and findings were noted and analysed. Type of study: Retrospective cross-sectional observational study. Objectives: Primary objective of the study was to assess the rates of laryngo-tracheal sequelae occurring in post-COVID-19 patients. Secondary objective was to assess the role of various factors along with COVID-19 that have led to the sequelae. Results: The study population included 348 participants, with an age distribution ranging from 21 to over 60 years. Post-COVID-19 symptoms were seen in 22.7% of all the patients included out of which hoarseness of voice 32.9%, difficulty in breathing 24.05%, and other related symptoms such as cough 18.98%, foreign body sensation 8.8%, throat irritation 7.5%, throat pain 5.06%, throat dryness 1.2%, and difficulty in swallowing 1.2% of the patients. Majority of our patients had history of hospital admission which was predominant (87.1%) over home isolation (10.3%). Most common comorbidity was diabetes mellitus (DM) (62.4%) along with hypertension. The various findings were noted in 10.1% of total patients included in the study including vocal cord polyps 19.9%, vocal cord paresis 19.9%, vocal cord granulomas 8.5%, vocal nodule 5.7%, and chronic congestion of laryngeal inlet 17.1% of the patients. Significant associations were observed between sequelae and factors such as hospital admission, intubation and tracheostomy, duration of O2 support, and diabetes mellitus. Conclusions: Laryngo-tracheal sequelae are commonly present and are persistent for months after recovery from COVID-19 in hospitalized and intubated/tracheostomized patients due to various factors related to the disease and its treatment. [ABSTRACT FROM AUTHOR]

Background: Newcastle disease significantly impacts the global poultry industry and is prevalent in many African countries, including Ethiopia. The objective of this research is to determine the humoral immune response to Newcastle Disease Virus (NDV), identify the circulating NDV genotype, and evaluate the correlation between the diagnostic tests used in backyard chickens in the Jimma Zone, southwest Ethiopia. Methods: A total of 90 swab and blood samples were purposively collected from symptomatic backyard chicken in the period between February and April 2022. Samples were collected from Jimma town, Seqa Chekorsa and Tiro Afeta districts of Jimma zone. Enzyme linked immunosorbent assay (ELISA) was conducted and seropositivity was determined from the collected serum samples. From the swab samples, total RNA was extracted and the viral genomic material was detected by amplifying the Fusion gene by Reverse transcription polymerase chain reaction (RT-PCR). The interconnection between ELISA and RT-PCR was also analyzed. Further, positive swab samples were nucleotide sequenced and genotyped. Results: Of the 90 serum samples, 62 (68.8%) were seropositive. From the 90 swab samples, 14 (15.5%) were RT-PCR positive. No statistically significant association between risk factors (breed, age, sex) and virus exposure was observed by RT-PCR (P = 0.41, 0.44, 0.67) or ELISA (P = 0.85, 0.19, 0.11). However, local breeds, young, and male birds were at higher risk according to RT-PCR results, while young and female birds were more likely to be seropositive. The antibody titer study showed that RT-PCR positive birds produced less than half the mean antibodies of negative birds (x̄=854 vs. 1885) and positive birds produced similar amount of antibody (σ = 626). Local (x̄=1978), adult (x̄=2558), and female (x̄=2620) birds had higher mean antibody titers than their counterparts. The agreement between RT-PCR and ELISA in identifying positive samples was minimal (k = 0.05). Nucleotide-sequenced isolates were nearly identical (99.7%) to each other and identified as velogenic based on the F-gene cleavage site (RRQKRF) with genotype VII.1.1. Conclusion: NDV was circulating in the study area, infecting birds with low antibody titers. High viral similarity between neighboring countries emphasizes the need for regional disease control strategy. [ABSTRACT FROM AUTHOR]

Sarcoma (SARC), a diverse group of stromal tumors arising from mesenchymal tissues, is often associated with a poor prognosis. Emerging evidence indicates that senescent cells within the tumor microenvironment (TME) significantly contribute to cancer progression and metastasis. Although the influence of senescence on SARC has been partially acknowledged, it has yet to be fully elucidated. In this study, we revealed that senescence level and age were associated with TME, immune treatment indicators, and prognosis in SARC. Utilizing the weighted gene co-expression network analysis and least absolute shrinkage and selection operator algorithm, we identified senescence-related genes and developed a senescence predictor. Three genes, RAD54, PIK3IP1, and TRIP13, were selected to construct a multiple linear regression model. Validation cohorts, immunohistochemistry, and quantitative reverse transcription polymerase chain reaction confirmed that the predictor derived from these three genes possessed prognostic and pathological relevance. Our senescence predictor provides comprehensive insights into the molecular mechanisms of SARC and identifies potential biomarkers for prognosis, paving the way for effective treatments. The results of this study hold promise for developing therapeutic strategies tailored to the unique characteristics of SARC. [ABSTRACT FROM AUTHOR]

Background: It is known that viral infections are associated with increased morbidity and mortality in immunocompromised children. Few reports describe the clinical manifestations and outcomes of COVID-19 infection in children and adolescents with benign and malignant hematological disease with a large sample size from our Egyptian population. Aim: As a preliminary step in developing a national registry for children and adolescents with hemato-oncological disease who were diagnosed with COVID-19 infection, in this study we aimed to determine (a) the frequency of COVID-19 infection in children with hematological disease, (b) the presenting clinical manifestations, and (c) the associated underlying disease outcome and its clinical determinants. Methods: We collected data from medical records of a retrospective cohort including all children and adolescent patients who were known to have a benign or malignant hematological disease attending the hematology and oncology outpatient clinic or were hospitalized between February 2022 and March 2023 in a university hospital in Cairo, Egypt, for tertiary care. All patients (n = 620) were tested for SARS-CoV-2 reverse transcription polymerase chain reaction (RT-PCR) test by nasopharyngeal swab. For all patients, the collected data included the demographic data, diagnosis, cause of hospital visit, cause and duration of admission if needed, presence of symptoms suggestive of COVID-19 infection, and concurrent medications. Baseline laboratory investigation were collected included complete blood count (CBC), liver and kidney functions, C-reactive protein (CRP), serum ferritin, lactate dehydrogenase, and coagulation test. For COVID-19-positive patients, the additional data collected included the duration of COVID-19 infection, its course, full examination during this illness, its severity, and the outcome of COVID-19 infection and the underlying disease. Results: Out of the 620 children and adolescents with benign (n = 271) and malignant (n = 349) hematological diseases in in-patients and out-patients' settings, 86 patients (13.8%) tested positive for SARS-CoV-2. COVID-19 infection was more common in patients with hematological malignancy (n = 56, 65.1%) than those with benign hematological disease (27 patients, 31.4%). Among COVID-19-positive patients, 20 patients (23.3%) required intensive care unit admission compared to 30 (5.6%) COVID-19-negative patients (p-value < 0.000). Death was reported in 5 COVID-19-positive patients (5.8%) compared to 19 in COVID-19-negative patients (3.6%) (p-value = 0.314). The factors associated with mortality in COVID-19-positive patients (3 patients with hematological malignancy and 2 patients with aplastic anemia) was abnormal neurological examination [OR = 17.3; p-value = 0.009]. Conclusion: The current study showed that COVID-19 was higher among patients with hematological malignancies than those with benign hematological disease. COVID-19 was tolerable among the studied patients with underlying hematological disease where 94.1% of the COVID-19-positive patients survived. Mortality was associated with abnormal neurological examination in COVID-19-positive patients with underlying hematological disease. [ABSTRACT FROM AUTHOR]

Background: Cytokeratins are intracellular proteins known as diagnostic biomarkers or prognostic factors for certain cancers. Cytokeratin 19 (CK-19) expression has been proven to have prognostic value for some cancers, but its relationship with others, such as prostate cancer (PCa), remains unclear. This systematic review article aimed to examine the relationship between CK-19 expression and prostate adenocarcinoma (PAC). Methods: To include the eligible studies that detected CK-19 expression in PAC, published articles since June 2024 were found using PubMed, Scopus, and Web of Science databases. The "prostate cancer" and "cytokeratin 19" keywords and their Mesh term were used for search databases. Data from the included articles were extracted and tabulated. This study was performed using the PRISMA guidelines, and the JBI checklist was used for the quality assessment. The study protocol was registered in PROSPERO under the "CRD42023472637" code. Results: Twenty-one studies were included. Eleven studies used reverse transcription polymerase chain reaction (RT-PCR) to investigate CK-19 expression, four used immunohistochemistry (IHC) staining, three used both one-step nucleic acid amplification (OSNA) and hematoxylin and eosin (H&E) methods, and three used the electrochemiluminescence (ECL) technique. CK-19 expression was detected in 301 patients among 619 patients. Additionally, only five out of 80 healthy donors were positive for CK-19 expression. Conclusion: Available evidence indicates a correlation between CK-19 expression and PAC progression, with higher CK-19 levels associated with advanced stages and worse prognosis. The overall evidence suggests that CK-19 could serve as a diagnostic and prognostic marker in PAC. [ABSTRACT FROM AUTHOR]

Background: LINC00312 has shown to play a suppressive role in the development and progression of non-small cell lung cancer (NSCLC). However, the expression pattern and diagnostic role of circulating LINC00312 in NSCLC remain to be confused. Methods: A total of 319 patients diagnosed with NSCLC and 180 healthy volunteers were enrolled from the First Affiliated Hospital of Huzhou University between January, 2022 and December, 2023. The participates were randomly assigned into the training and validation groups with a ratio of 6:4, while the remaining was named as the exosomal group. Reverse transcription-quantitative PCR (RT-qPCR) was employed to investigate the expression pattern of LINC00312 in NSCLC tissues, serum samples and cell lines. Receiver operating characteristic (ROC) curve analysis was carried out for distinguishing NSCLC from healthy volunteers. Results: Here, we revealed that LINC00312 was lowly expressed in NSCLC and low LINC00312 expression manifested a poor prognosis. Additionally, compared with the healthy volunteer group, a reduction of circulating LINC00312 in patients with NSCLC was observed in both the training and validation groups. Further correlation analysis indicated that circulating LINC00312 expression was tightly associated with lymph node metastasis, cancer thrombus, spread through air space (STAS) status and pathological type. Moreover, circulating LINC00312 showed a good performance to distinguish NSCLC from healthy volunteers with a higher sensitivity and specificity values. Lastly, exosomal LINC00312 level was also decreased in NSCLC compared with in healthy volunteers. Conclusions: Taken together, these data unveil that circulating LINC00312 was notably downregulated in NSCLC, offering a novel non-invasive marker for diagnosis of NSCLC. [ABSTRACT FROM AUTHOR]

Background: We examined chronic gadolinium retention impact on gene expression in the mouse central nervous system (CNS) after injection of linear or macrocyclic gadolinium-based contrast agents (GBCAs). Methods: From 05/2022 to 07/2023, 36 female mice underwent weekly intraperitoneal injections of gadodiamide (2.5 mmol/kg, linear), gadobutrol (2.5 mmol/kg, macrocyclic), or saline. Mice were sacrificed on day 29 or 391 after a 1-year washout. Assessments included magnetic resonance imaging (MRI), mechanical hyperalgesia tests, and inductively coupled plasma mass spectrometry to measure gadolinium levels. Ribonucleic acid (RNA) sequencing and bioinformatic analyses identified differentially expressed genes (DEGs), with validation by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB). Results: Post-gadodiamide, MRI showed increased signal intensity in the deep cerebellar nuclei (pre, 0.997 ± 0.006 versus post, 1.086 ± 0.013, p < 0.001). Mechanical hyperalgesia tests indicated transient sensory changes. After 1-year, gadolinium retention was noted in the brain (5.92 ± 0.32 nmol/kg) and spinal cord (1.23 ± 0.66 nmol/kg) with gadodiamide, compared to saline controls (0.06 ± 0.02 nmol/kg in brains and 0.28 ± 0.06 nmol/kg in spinal cords). RNA sequencing identified 17 shared DEGs between brain and spinal cord in the gadodiamide group on day 391, with altered Hmgb2 and Sgk1 expression confirmed by qRT-PCR and WB. Reactome pathway analysis showed enrichment in neuroinflammation pathways. No DEGs were detected in brains on day 29. Conclusion: Chronic gadolinium deposition from repeated linear GBCA but not macrocyclic administration causes significant gene expression alterations in the mouse CNS, particularly affecting neuroinflammation pathways. Relevance statement: This study examined the long-term impact of chronic gadolinium retention on gene expression in the mouse CNS, uncovering significant changes associated with neuroinflammation pathways after repeated administration of linear GBCA, but not with macrocyclic GBCA. These findings highlight the importance of further research on the long-term safety of linear GBCA in medical imaging. Key Points: Chronic gadolinium retention alters gene expression in the mouse central nervous system. Significant neuroinflammatory pathway changes were observed after linear gadodiamide exposure. MRI showed increased signal intensity in deep cerebellar nuclei after gadodiamide injection. [ABSTRACT FROM AUTHOR]

Background: Triple-negative breast cancer (TNBC) is among the most aggressive forms of breast cancer, characterized by a dismal prognosis. In the absence of drug-targetable receptors, chemotherapy remains the sole systemic treatment alternative. Recent advancements in immunotherapy, particularly immune checkpoint inhibitors (ICIs) that target programmed death 1/programmed death ligand 1 (PD-1/PD-L1) and cytotoxic T lymphocyte associated antigen 4 (CTLA-4), have provided renewed optimism for the treatment of patients with TNBC. Prior research has indicated that the expression level of the cell polarity protein discs large homolog 5 (DLG5) correlates with the malignant progression and prognosis of breast cancer; nevertheless, its influence on PD-L1 expression and its function in immunotherapy for TNBC require further investigation. Methods: The hypoxia cell model was established by simulating the cell hypoxic microenvironment in the human SUM159 and MDA-MB-231 cell lines using cobalt II chloride (CoCl2). A combination of PD-L1 inhibitors and DLG5 RNA interference techniques was used, along with various methods including cell counting kit-8 (CCK-8), colony formation, wound healing, transwell migration, reverse transcription-quantitative real-time PCR (RT-qPCR), immunofluorescence, immunohistochemical staining (IHC), expression analysis from datasets and western blotting. These methods were employed to evaluate changes in cell proliferation, migration, and the expression levels of PD-L1 and DLG5. Additionally, the correlation between the expression of PD-L1 and DLG5 in clinical samples was analyzed. Results: (1) In vitro experiments, a cellular hypoxia model was effectively established utilizing 150 µM CoCl₂. Under these conditions, cell clone formation, invasiveness, and migration rate were all significantly inhibited. (2) The expression levels of DLG5 and PD-L1 were significantly increased in both MDA-MB-231 and SUM159 cells following treatment with 150 µM CoCl₂. (3) Silencing DLG5 resulted in a considerable upregulation of PD-L1 expression in MDA-MB-231 and SUM159 cells under normoxic circumstances, but it was markedly downregulated under hypoxic settings. Inhibition of PD-L1 expression resulted in a considerable increase in DLG5 expression under normoxic conditions, but it decreased under hypoxic conditions. Correlation research demonstrated an inverse association between the expression of DLG5 and PD-L1 in TNBC tissues. Conclusion: This study provides new theoretical evidence and potential therapeutic targets for the immunotherapy strategies of TNBC, holding significant clinical application value. [ABSTRACT FROM AUTHOR]

Objective: Despite N6‐methyladenosine (m6A) being closely involved in various pathophysiological processes, its potential role in liver injury is largely unknown. We designed the current research to study the potential role of fat mass and obesity‐associated protein (FTO), an m6A demethylase, on hepatic ischemia‐reperfusion injury (IRI). Methods: Wild‐type mice injected with an adeno‐associated virus carrying fat mass and obesity‐associated protein (AAV‐FTO) or adeno‐associated virus carrying green fluorescent protein (GFP) (AAV‐GFP) were subjected to a hepatic IRI model in vivo. Hematoxylin–eosin staining was performed to observe IRI. Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining was used to observe the cell apoptosis. Reverse transcription polymerase chain reaction (RT‐PCR) was used to observe the expression of FTO. The protein levels of FTO, apoptosis, or autophagy‐associated signaling proteins were detected by western blot. Reactive oxygen species (ROS) levels were determined by flow cytometry, and immunohistochemistry was used to detect the FTO and LC3‐II expression. For in vitro experiments, cultured hepatocytes were subjected to hypoxia/reoxygenation (H/R) stimulation. Monodansylcadaverine (MDC) staining was used to visualize autophagic vesicles. Results: In the present study, we showed that FTO was involved in hepatic IRI, apoptosis, and autophagy. Specifically, the expression level of FTO was significantly reduced in the hepatic IRI. Besides, increasing FTO expression (AAV‐FTO) ameliorated the hepatic IRI in animal models, accompanied by decreased apoptosis and autophagy. Furthermore, the FTO inhibitor (FB23‐2) aggravated autophagy in hepatocytes upon H/R‐induced damage. Conclusion: FTO could act as a protective effector during hepatic IRI, associated with decreased apoptosis and autophagy. FTO‐mediated m6A demethylation modification may be an important therapeutic target for hepatic IRI. [ABSTRACT FROM AUTHOR]

Background: Low-intensity pulsed ultrasound (LIPUS) has been used as an effective noninvasive method for treating fractures and osteoarthrosis, but the application in the field of oral implantation is in its infancy. This study aimed to clarify the effect and mechanism of LIPUS on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and implant osseointegration, and to provide an experimental basis for future clinical applications. Methods: Dental implants were inserted into Wistar rat femurs, and LIPUS was performed for 4 weeks. Micro-CT and toluidine blue staining were used to assess implant osseointegration. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were used to identify enriched functional terms and signalling pathways for differentially expressed genes from LIPUS-treated rat BMSC RNAseq data obtained from the GEO database. The random forest method was used to identify key risk genes according to the mean decrease Gini (MDG) coefficient. Then, LIPUS was applied to treat rat BMSCs, and alkaline phosphatase (ALP) staining, alizarin red staining, RT-PCR and western blotting were used to determine whether LIPUS could promote BMSC osteogenic differentiation via integrin α11 (ITGA11) and the focal adhesion pathway. Results: Our in vivo experimentations verified that LIPUS significantly increased new bone formation and osseointegration around the implant in rats. Bioinformatics analysis of RNA-seq data revealed that the upregulated genes in BMSCs after LIPUS treatment were significantly enriched in osteoblast differentiation-related functions and focal adhesion-related pathways. Random forest analysis revealed that ITGA11 was the most significant factor affecting BMSC osteogenic differentiation among the differentially expressed genes. In addition, LIPUS significantly increased ALP expression and mineralized nodule formation in rat BMSCs by upregulating ITGA11 and increasing the activity of FAK/PI3K/AKT/GSK3β/β-catenin pathway. Conclusions: LIPUS can effectively promote implant osseointegration in rats and improve rat BMSC osteogenic differentiation by upregulating ITGA11 and increasing the activity of the downstream focal adhesion pathway. [ABSTRACT FROM AUTHOR]

Yellow nail syndrome is a rare disorder characterized by the triad of yellow dystrophic nails, lymphedema, and chronic lung disease. Most cases occur in adulthood, with only a few congenital or familial cases reported. The pathogenesis of yellow nail syndrome remains largely unknown. This case series, which includes both congenital and sporadic cases, aimed to elucidate the key mechanistic pathways underlying the development of this disease. Background: Yellow nail syndrome (YNS) is a rare disorder characterized by a triad of yellow dystrophic nails, lymphedema, and chronic lung disease. Most patients present in adulthood, with only a few congenital or familial cases described. The cause of YNS remains largely unknown, although defects in lymphatic vessel development are suggested to play a significant role. Objective: To elucidate the genetic mechanisms underlying YNS. Design: Analysis of genetic sequencing data and gene and protein expression studies. Setting: A tertiary care academic medical center. Patients: 6 patients with congenital YNS (cYNS) and 5 with sporadic YNS (sYNS). Measurements: Exome and genome sequencing were used to detect disease-causing variants, complemented by RNA analyses for intronic variants. Protein and gene expressions were studied by immunofluorescence staining and real-time reverse transcriptase quantitative polymerase chain reaction analyses. Results: Biallelic variants in CELSR1 (n  = 5) or likely FZD6 (n  = 1), both core molecules in the Wnt/planar cell polarity (PCP) pathway, were identified in all patients with cYNS; none of the patients with sYNS had candidate genetic variants. Immunofluorescence staining showed that CELSR1 colocalizes with lymphatic vessels in the skin but not in the lungs or the intestine. Moreover, levels of CELSR1 and FZD6 proteins were negligible to zero in patient tissues (n  = 2) compared with control tissues. Gene expression of Wnt/PCP–related genes was reduced in patients with cYNS (n = 3), and patients with sYNS (n  = 4) showed milder gene expression impairments. Limitation: Small cohort size and limited sample availability. Conclusion: Defects in PCP organization may play a major role in the pathogenesis of YNS. To the authors' knowledge, this is the first demonstration of a mechanism explaining YNS development, mainly in its congenital form but also in patients with sporadic disease. Primary Funding Source: The Prof. Baum Research Fund of Israel Lung Association. [ABSTRACT FROM AUTHOR]

Purpose: The present study investigated the role of resistance-nodulation-cell division (RND) efflux pumps in tigecycline resistance of Acinetobacter baumannii clinical isolates recovered from three Western Balkan countries (Serbia, Bosnia and Herzegovina and Montenegro). Methods: A total of 37 A. baumannii isolates recovered from seven tertiary care hospitals in 2016 and 2022 were tested against tigecycline using broth microdilution method. Then, efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone (CCCP) was used to determine the involvement of efflux pumps in tigecycline resistance. Molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multiplex PCR-based determination of clonal lineage. Regulators of efflux pumps were analyzed for amino acid substitutions, while reverse transcription-quantitative PCR (RT-qPCR) enabled quantification of RND efflux pumps expression. Results: All tested isolates were interpreted as resistant to tigecycline and showed reduced tigecycline minimum inhibitory concentration (MIC) values in the presence of CCCP. PFGE analysis showed significant diversity among isolates grouped in cluster I including IC2 (n = 32) and IC3 (n = 1) isolates, while cluster II was comprised of four IC1 isolates. The most prevalent substitutions in AdeR were V120I and A136V and in AdeS G186V and N268H (n = 33). The Q262R substitution was detected in AdeL proteins of IC1 isolates, whereas no alterations were observed within AdeN. The expression of the adeB, adeG, and adeJ genes in selected isolates was upregulated in five (1.16- to 3-fold), sixteen (1.35- to 2.82-fold), and twelve isolates (1.62- to 4-fold) compared to ATCC19606, respectively. Conclusion: This study revealed that overexpression of RND efflux pumps underlies tigecycline resistance in A. baumannii clinical isolates from the Western Balkans. [ABSTRACT FROM AUTHOR]

Objectives: An increasing body of evidence suggests that N6-methyladenosine (m6A) plays a crucial role in the etiology of SLE. We focused on YTHDF2 expression in neutrophils and the relationship between YTHDF2 and the pathogenesis of SLE. Methods: Sixty-one patients with SLE and 48 healthy controls (HC) were recruited, and their clinical characteristics were recorded. The mRNA levels of m6A "writers" (METTL3, METTL14, WTAP), "erasers" (FTO and ALKBH5), "readers" (YTHDF2), and inflammatory factors (interleukin-1β, interleukin-6, interleukin-8, and TNF-α) in neutrophils were determined by reverse transcription-quantitative PCR. The protein of YTHDF2 was determined by Western blotting. The correlations between the YTHDF2 in neutrophils of SLE patients and clinical features were examined by Spearman's method. YTHDF2 and TNF-α expression in neutrophils were examined after stimulation by SLE serums. Results: The mRNA expression of YTHDF2 in neutrophils was significantly decreased, and the protein level of YTHDF2 in neutrophils was decreased. The mRNA expression of YTHDF2 in neutrophils correlated with L% (rs = 0.5796, P < 0.0001), LMR (rs = 0.3524, P = 0.0062), WBC (rs = − 0.3186, P = 0.0123), N (rs = − 0.4141, P = 0.0010), N% (rs = − 0.4813, P < 0.0001), NLR (rs = − 0.5301, P < 0.0001), dNLR (rs = − 0.4945, P < 0.0001), and SII (rs = − 0.3930, P = 0.0019), which were suggested as the inflammatory conditions of SLE. In addition, the mRNA expression of TNF-α in neutrophils was significantly increased in SLE patients. Further analysis revealed that the mRNA expression of YTHDF2 in neutrophils was inversely correlated with TNF-α in SLE. Neutrophils from health control were significantly downregulated in their YTHDF2 expression and upregulated in their TNF-α expression following stimulation by serum from SLE patients. Conclusion: This study indicates that the levels of YTHDF2 in peripheral blood neutrophils may be involved in the pathogenesis of SLE and could be a novel target for diagnosis and therapy. Key points • The mRNA expression of YTHDF2 in neutrophils of SLE and described that decreased mRNA of YTHDF2 in neutrophils correlated with L%, LMR, WBC, N, N%, NLR, dNLR, and SII. • The mRNA expression of TNF-α in neutrophils was significantly increased and correlated with YTHDF2 in SLE patients. • Neutrophils from health control were significantly downregulated in their YTHDF2 expression and upregulated in their TNF-α expression following stimulation by serum from SLE patients. [ABSTRACT FROM AUTHOR]

Previous studies have shown that oral whole honey reduces weight gain in rats on a normal diet (ND) or high-fat diet (HFD) and suppresses inflammation by modulating immunological cytokines in human neutrophils and macrophages. We hypothesize that the honey proteins (HP) are responsible for the reduced weight gain in rats on ND and HFD and that HP would alleviate obesity parameters. To test this, proteins were isolated from acacia honey through the salting-out method. Wistar rats (N = 24) were randomized to get ND or HFD for 4 weeks, then further randomized to four groups and treated with HP or saline for another 4 weeks. Energy intake (EI), body weight gain, EI per gram body weight gain, serum glucose, and lipids were measured. Expression of adipose tissue genes fatty acid binding protein (FABP), lipase C (LIPC), and apolipoprotein A-1 (APOA1) was evaluated through the quantitative polymerase chain reaction. HFD increased the body weight versus ND in weeks 1–4. HP for the next 4 weeks reduced weight gain in ND-HP and HFD-HP groups versus saline controls (P <.01). EI was not significantly different among groups. However, EI per gram body weight gain among groups was markedly different (P <.01), demonstrating reduced weight gain efficiency by HP (P <.01). HP reduced glucose in ND but not in HFD groups. Triglycerides were lower in both HP groups. The expressions of FABP, LIPC, and APOA1 genes were significantly increased (P <.05) in HP-treated HFD rats. Collectively, weight gain efficiency was remarkably reduced without altering EI in rats following the HP treatment, suggesting HP increased metabolic rate or substrate partitioning. Studies of HP are suggested in humans. [ABSTRACT FROM AUTHOR]

Background: Magnolia kobus DC (MO), as a plant medicine, has been reported to have various physiological activities, including neuroprotective, anti-inflammatory, and anti-diabetic effects. However, vascular protective effects of MO remain incompletely understood. In this study, we evaluated the vascular protective effect of MO against ferroptosis in a carotid artery ligation (CAL)-induced neointimal hyperplasia mouse model and in aortic thoracic smooth muscle A7r5 cells. Methods: This study was conducted to estimate the vascular protective effects of MO by systematically measuring histopathological analysis and western blot analysis in CAL animal model. In vitro protective effects of MO were evaluated by estimating cell viability, reactive oxygen species (ROS) content, glutathione (GSH) levels, lipid peroxidation, mitochondrial morphological change, cell proliferation, migration, western blot analysis, and qRT-PCR against erastin (Era)-induced A7r5 cells. Results: MO intake significantly improved neointimal formation, inhibited ferroptosis and vascular smooth muscle cell (VSMC) phenotypes, and ameliorated the antioxidant system of carotid artery tissues. In addition, MO treatment effectively ameliorated Era-induced ferroptotic cytotoxicity, including cellular death, ROS production, and cell migration status. MO treatment also suppressed proliferation and migration in Era-induced A7r5 cells. MO considerably regulated Era-induced abnormal mechanisms related to ferroptotic changes, VSMC phenotype switching, and the ROS scavenging system in A7r5 cells. Conclusion: MO has the potential for use as a functional food supplement, nutraceutical, or medicinal food, with protective effects on vascular health by regulating ferroptosis and VSMC phenotypic switching. [ABSTRACT FROM AUTHOR]

The article in the International Journal of Laboratory Hematology discusses a case of acute myeloid leukemia with a cryptic KMT2A::MLLT10 rearrangement, which was not initially detected by standard testing methods. The patient had an aggressive clinical course with unfavorable abnormalities. Molecular techniques, such as RT-MLPA and NGS on RNA, were crucial in identifying the rearrangement. The study emphasizes the importance of accurate genomic testing for proper disease classification and patient risk assessment in cases of AML with complex genetic abnormalities. [Extracted from the article]

Background: Conflicting guidance exists regarding the characterization of nebulization as an aerosol-generating procedure and subsequent risk of transmission of SARS-CoV-2 to healthcare workers. Aim/Objective: This study sought to address whether SARS-CoV-2 viral load distribution was impacted by bronchodilator delivery via nebulizer versus metered-dose inhaler (MDI). Methods: Adults infected with COVID-19 were enrolled and received a single dose of albuterol sulfate nebulized solution (2.5 mg/3 mL via breath-actuated nebulizer with filtered mouthpiece) or albuterol sulfate hydrofluoroalkane inhalation aerosol (90 µg/actuation; two puffs via MDI with spacer) in a randomized crossover sequence. Air and surfaces were sampled at predefined locations within patients' hospital rooms to assess SARS-CoV-2 dispersion over three periods (baseline, MDI, and nebulizer). Findings/Results: Eleven patients received crossover therapy. Six patients had detectable SARS-CoV-2 RNA during one treatment period (n = 3 each for MDI and nebulizer) and one during both treatment periods. No viral RNA was found in the rooms of four (36.4%) patients. Overall, few environmental samples (17/397; 4.3%) contained detectable viral RNA, with no meaningful differences in positivity rate across periods; RNA genome copy numbers were low in positive samples. No correlation between dispersion and patient clinical status or environmental parameters was observed. Discussion: In this first prospective trial evaluating viral load distribution following use of nebulizer versus MDI in hospitalized patients with COVID-19, low environmental contamination was found regardless of administration method. Findings support the use of either device when needed to treat patients with COVID-19. [ABSTRACT FROM AUTHOR]

Background: Breast cancer (BC) is the most prevalent solid cancer affecting women's health globally. Matrine (MAT), a traditional Chinese herb, has exhibited antitumor effects against BC. However, its mechanism of action, particularly whether it involves the control of cell proliferation and epithelial-mesenchymal transition (EMT), remains unknown. Aims: To explore MAT's role in BC and its regulatory mechanisms, as well as to identify targets for the development of novel medicines and improvement of BC treatment modalities. Study Design: Experimental study. Methods: The UALCAN and Lnc2Cancer 3.0 databases were used to predict the expression of LINC01116 in BC. The BC cells (MDA-MB-231 and MCF-7) were treated with various concentrations of MAT, and the optimal dose and timing of MAT action were determined using CCK-8 and quantitative real-time polymerase chain reaction assays. Functional assays such as CCK-8, EdU, Transwell, Western blot, and flow cytometry assays were performed on the BC cells, and the impacts of LINC01116, miR-9-5p, and ITGB1 expression levels on MAT's mechanism of action were assessed. The association between LINC01116, miR-9-5p, and ITGB1 was evaluated using dual luciferase and RNA immunoprecipitation assays. Furthermore, the size and weight of the subcutaneous tumors in mice model were assessed. The effect of LINC01116 overexpression on the in vivo action of MAT and histopathological staining (TUNEL immunofluorescence, hematoxylin & eosin staining, immunohistochemistry staining for Ki67 and Bax) were also assessed. Results: The optimal dose and duration of MAT administration were 8 µm and 24 h, respectively. MAT effectively inhibited BC cell proliferation, EMT progression, and biological functions, while promoting BC cell apoptosis. The animal model experiments also demonstrated that MAT inhibited BC tumor growth in vivo. Furthermore, MAT inhibited LINC01116, which acted as a sponge for miR-9-5p, increasing the ITGB1 level. Conclusion: MAT suppresses BC cell and EMT proliferation via the LINC01116/miR-9-5p/ITGB1 pathway. Thus, MAT may be a promising target for adjuvant anti-BC therapy. [ABSTRACT FROM AUTHOR]

Continued heat exposure can cause physiological and cellular responses. This study investigated the association between physiological responses and heat shock protein 70 (HSP70) expressions in Kuala Lumpur's urban vulnerable population. We conducted a cross-sectional study involving 54 participants from four areas classified as experiencing moderate to strong heat stress. Physiological measurements included core body temperature, heart rate, and diastolic and systolic blood pressure. RT-qPCR and ELISA were also performed on blood samples to assess HSP70 gene and protein expressions. Despite indoor heat stress, participants maintained normal physiological parameters while there were significant indications of HSP70 expression at both the gene and protein levels. However, our study found no significant correlation (p > 0.05) between physiological responses and HSP70 expressions. This study shows no interaction between physiological responses and HSP70 expressions in the study population, revealing the complex mechanisms of indoor heat stress in vulnerable individuals. [ABSTRACT FROM AUTHOR]

Zika fever is commonly perceived as a mild illness; however, often is linked to congenital Zika syndrome in newborns and Guillain–Barré syndrome in adults. The etiological agent, Zika virus, is a globally distributed arbovirus, frequently prevalent in tropical and humid climates. Conventional laboratory identification relies on real-time reverse transcription polymerase chain reaction (RT-PCR), a standard method requiring specialized infrastructure and technical training. To address these challenges, this research introduces the development of a nano immunosensor based on gold nanoparticles (AuNPs), for Zika virus biorecognition, assessed through Fourier-transform infrared spectroscopy (FTIR). The study further identifies signals associated with specific functional groups of viral envelopes. Ultra-high-resolution scanning electron microscopy (SEM) demonstrates the biorecognition of viral particles. Given the morphological, biochemical, and immunogenic resemblance between Zika and dengue viruses, this nano immunosensor is proficient in detecting dengue virus as well. Moving to the analysis phase, FTIR signals obtained after nano immunosensor–virus interaction were processed by principal component analysis (PCA). This achieved adequate discrimination between both viruses in key spectral regions: phospholipids, esters, and glycerides (1755–1726 cm−1), proteins (1683–1500 cm−1, 1328–1275 cm−1), and carbohydrates (1134–993 cm−1). These results underscore the utility of the AuNP-based nano immunosensor for identifying and discriminating between Zika and dengue viruses, offering a viable alternative in the diagnosis of these pathogenic agents. [ABSTRACT FROM AUTHOR]

Pyoderma gangrenosum (PG) is a rare, chronic, ulcerative disease characterized by non-healing wounds that worsen with debridement, a phenomenon called pathergy. No consensus regarding pathogenesis, diagnosis, or treatment exists for PG. A previous pilot study using dehydrated human amniotic/chorionic membrane (dHACM), following excisional debridement, augmented PG wound healing and allowed for subsequent wound closure through split-thickness skin grafting (STSG). In this clinical trial (NCT05120726), four patients with an established PG diagnosis were enrolled to undergo treatment with dHACM and characterize the pre- and post-treatment transcriptome profiles. RNA sequencing was used to isolate the total RNA from specimens. Genes of particular interest were quantified through real-time quantitative reverse transcription polymerase chain reaction. We observed varied changes to the local expression of inflammatory response, positive regulators of cellular proliferation, and extracellular matrix disassembly cytokines. All PG wounds produced granulation tissue following treatment and were closed using split-thickness skin grafts. [ABSTRACT FROM AUTHOR]

Regulatory T cells (Tregs) ameliorate inflammatory bowel diseases. However, their plasticity is not completely understood. In this study using a mouse colitis model, Tregs and T helper 17 (Th17)-like Tregs were detected and sorted using flow cytometry, followed by transcriptome sequencing, real-time reverse transcription polymerase chain reaction, and flow cytometry to analyze the mRNA profiles of these cells. Treg plasticity was evaluated by in vitro differentiation assays. The immunosuppressive activities of Tregs and Th17-like Tregs were assessed in an adoptive transfer assay. We found Treg-derived Th17-like Tregs in inflamed colonic lamina propria (LP). LP Th17-like Tregs expressed higher Th17-related cytokines and lower immunosuppressive cytokines compared with LP Tregs. Notably, Tregs expressed higher Yes-associated protein 1 (YAP1) but lower transcriptional coactivator with PDZ-binding motif (TAZ) than Th17-like Tregs. Verteporfin-mediated inhibition of YAP1 activity enhanced Th17-like Treg generation, whereas IBS008739-induced TAZ activation did not affect Th17-like Treg generation. Besides, verteporfin enhanced while IBS008739 suppressed the differentiation of Th17-like Tregs into Th17 cells. Furthermore, YAP1 activated STAT5 signaling in Tregs, whereas YAP1 and TAZ activated STAT3 and STAT5 signaling in Th17-like Tregs. Compared with Tregs, Th17-like Tregs were less efficacious in ameliorating colitis. Therefore, YAP1 suppressed Treg differentiation into Th17-like Tregs. Both YAP1 and TAZ inhibited the differentiation of Th17-like Tregs into Th17 cells. Therefore, YAP1 and TAZ probably maintain the immunosuppressive activities of Tregs and Th17-like Tregs in colitis. [ABSTRACT FROM AUTHOR]

Deranged lipid homeostasis has been implicated in neurodegenerative diseases. Cholesterol reducing compounds such as statins have received special attention for the possibility that they may be able to ameliorate or prevent cognitive loss associated with neurodegeneration. However, there is much dissension concerning the actual effect of statins on cognitive function. The aim of this study is to investigate the effects of pitavastatin on hippocampal synaptogenesis because the hippocampus is crucial for memory formation. We also evaluated the effects of pitavastatin on local hippocampal estrogen synthesized in the hippocampus itself and its effect on Brain-Derived Neurotrophic Factor (BDNF). Using a hippocampal cell line, H19-7, we found that hippocampal neurons exposed to pitavastatin demonstrate a significant reduction in the synaptic marker postsynaptic density protein 95 (psd-95). The pitavastatin treated neurons also exhibited decreased production of local estrogen and their expression of BDNF mRNA was decreased. These results suggest that statins reduce the ability of hippocampal neurons to form synapses by restricting the production of local estrogen. Because neural connections in the hippocampus are crucial for memory formation, our findings implicate statins as medications that may compromise cognitive function. [ABSTRACT FROM AUTHOR]

This study examines the role of N-acetylglucosamine kinase (NAGK) in initiating puberty in female mice. We employed real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and immunofluorescence to measure NAGK expression in the hypothalamic-pituitary-ovarian axis across various developmental stages: infant, prepuberty, puberty, and adult. We further investigated the impact of Nagk gene knockdown on puberty in female mice. This included assessing the expression of puberty-related genes both in vivo and in vitro , GT1-7 cells proliferation and apoptosis, concentrations of GnRH and Kisspeptin, puberty onset timing, serum levels of progesterone (P 4) and estradiol (E 2), and ovarian morphology. Results revealed that Nagk mRNA is present in the hypothalamus, pituitary, and ovaries throughout different developmental stages in female mice. In the hypothalamus, Nagk mRNA levels were comparable during infant and prepuberty, lowest during puberty, and highest in adult. In the pituitary, Nagk mRNA peaked in adult, with no significant variation between infant, prepuberty, and puberty. In the ovaries, Nagk mRNA levels increased during puberty and peaked in adult. NAGK is predominantly located in the arcuate nucleus (ARC), periventricular nucleus (PeN), dorsomedial hypothalamic nucleus (DMH), paraventricular nucleus (PVN), adenohypophysis, and in the ovarian oocytes, interstitium, and granulosa cells across all developmental stages in female mice. Nagk knockdown in GT1-7 cells decreased the transcriptional level of Gnrh , Kiss1 , Gpr54 , Igf1 and Mapk1 4 mRNA and cell proliferation but increased the level of β-catenin mRNA and cell apoptosis, while reducing GnRH secretion. Following ICV injection, Nagk gene knockdown mice exhibited delayed the timing of vaginal opening (VO) and reduced hypothalamic levels of Gnrh , Kiss1 , Gpr54 , Igf1 , Mapk14 , and β-catenin mRNA. Additionally, serum concentrations of E 2 in Nagk gene knockdown mice were significantly lower compared to the control group. These findings indicate that Nagk regulates the expression of Gnrh and Kiss1 mRNA in GT1-7 cells, affects hypothalamus Gnrh mRNA levels and serum E 2 concentration, and that its knockdown can delay puberty onset in female mice. • NAGK was expressed in the reproductive axis of female mice, with mRNA and protein levels varying by developmental stages. • Knockdown of Nagk in GT1-7 cells can reduce the transcription levels of puberty-related genes. • Knockdown of Nagk in GT1-7 cells can inhibit GnRH secretion and cell proliferation, and promote apoptosis. • Knockdown of Nagk delayed the age at vaginal opening and reduced serum E 2 level in mice. [ABSTRACT FROM AUTHOR]

Bovine viral diarrhea virus (BVDV) is a productive and reproductive virus with a high global economic impact on dairy production systems. We investigated the prevalence of BVDV at the herd and individual levels in most dairy-producing regions of Brazil. The frequency of BVDV at the herd level was investigated using quantitative reverse transcription polymerase chain reaction (RT-qPCR) in bulk milk tanks monitored at 289 dairy farms between August 2020 and January 2022. Among these farms, 68 production systems were selected to investigate the prevalence of persistently infected (PI) animals using two antigen-enzyme-linked immunosorbent assays (AgELISAs), at 21-d intervals, from ear-notch samples. In total, 2,902 RT-qPCR and 23,466 Ag-ELISAs were performed. At the herd level, 23.87% (69/289) of dairy farms were considered infected, presenting at least one qPCR test positive. At the individual level, 41.2% (28/68) of the subgroup of selected farms had at least one animal positive in the Ag-ELISA test. Association between tests allowed the classification of farms into the following four categories: level 0, negative for both tests (41.2%, 28/68); level 1, RT-qPCR positive and Ag-ELISA negative (17.6%, 12/68); level 2, RT-qPCR negative and Ag-ELISA positive (13.2%, 9/68); and level 3, positive for both tests (27.9%, 19/68). Multiple correspondence analysis (MCA) suggested a possible association between BVDV positivity and large farms, average daily milk production (herd), breed, and somatic cell counts. The confinement and intensification of animals from different categories, and use of artificial bedding are associated with BVDV infection. Using waste or bulk tank milk to fed calves was also a risk factor for BVDV positivity in RT-qPCR and Ag-ELISA. Despite the use of reproductive vaccines by most producers, their use seems to be associated with BVDV-positive farms. This study presented the epidemiological frequencies of BVDV at the individual and herd levels in the Campos Gerais Paranaense region. The region ranks among the top milk-producing areas in Brazil. Additionally, the association between BVDV tests and farm characteristics indicated the farm risk for BVDV and guides specific control programs. [ABSTRACT FROM AUTHOR]

Heat stress is essential for improving the efficacy of mesenchymal stem cell (MSC)-based regeneration medicine. However, it is still unclear whether and how heat stress influences the differentiation of stem cells from apical papilla (SCAPs). This research aimed to explore the potential mechanism of glucose-regulated protein 78 (GRP78) in regulating differentiation under heat stress in SCAPs. The proliferation ability was assessed using the 5-Ethynyl-2'- deoxyuridine (EdU) assay, cell counting kit assay (CCK-8), and flow cytometry (FCM). The osteogenic and odontogenic differentiation capacities were investigated through Western blot, quantitative reverse transcription polymerase chain reaction (qRT-PCR), alkaline phosphatase (ALP) staining and activity assay, alizarin red S (ARS) staining, as well as immunofluorescence staining. Western blot and transmission electron microscopy (TEM) were used to detect autophagy. Heat stress enhanced the osteogenic and odontogenic differentiation of SCAPs, but it did not significantly affect proliferation. Besides, GRP78 has been confirmed to modulate the differentiation induced by heat stress. Autophagy triggered by GRP78 enhanced osteogenic and odontogenic differentiation of SCAPs, while the knockdown of GRP78 or the inhibitor of autophagy suppressed the differentiation. Heat stress induces osteogenic and odontogenic differentiation of SCAPs through GRP78-mediated autophagy. [ABSTRACT FROM AUTHOR]

A machine learning‐based approach was utilized to develop a device for groundnut bud necrosis virus (GBNV) disease severity detection and estimation in tomato plants (Solanum lycopersicum L.). The study involved inoculating tomato plants with GBNV, monitoring changes in morphological and spectral characteristics, evaluating machine learning algorithms (decision tree [DT] classifier) for analysis and classification of disease severity, and developing and validating a device for disease detection and severity estimation. Spectral data analysis revealed distinct patterns in reflectance, with notable peaks observed in the 680 and 760 nm bands, while reflectance remained low and constant beyond 900 nm. Machine learning techniques, specifically a DT model, were employed to classify disease severity based on spectral data with high accuracy (95.01% training accuracy and 93.65% testing accuracy). The model identified the near‐infrared band as highly correlated (correlation coefficient of 0.82) with disease severity. Furthermore, a compact handheld device integrating a spectral sensor, organic light‐emitting diode display, and Raspberry Pi 3B was developed for real‐time disease severity estimation. The device demonstrated robust performance, accurately predicting disease severity at different growth stages, even in the absence of visible symptoms. Additionally, disease severity percentages obtained via reverse transcription polymerase chain reaction were used to validate the accuracy of the device's estimations. Its responsive nature, with estimated response times ranging from milliseconds to seconds, facilitates timely interventions in agricultural settings. Overall, this interdisciplinary approach, combining spectral analysis, machine learning, and device development, presents a promising solution for efficient disease monitoring and management in agriculture, contributing to enhanced crop health and food security. [ABSTRACT FROM AUTHOR]

Objective: To investigate the therapeutic effect of Xiangshao Granules (XSG) on post-stroke depression (PSD) and explore the underlying mechanisms. Methods: Forty-three C57BL/6J mice were divided into 3 groups: sham (n=15), PSD+vehicle (n=14), and PSD+XSG (n=14) groups according to a random number table. The PSD models were constructed using chronic unpredictable mild stress (CUMS) after middle cerebral artery occlusion (MCAO). The sham group only experienced the same surgical operation, but without MACO and CUMS stimulation. The XSG group received XSG (60 mg/kg per day) by gavage for 4 weeks. The mice in the sham and vehicle groups were given the same volume of 0.9% saline at the same time. The body weight and behavior tests including open field test, sucrose preference test, tail suspension test, and elevated plus-maze test, were used to validate the PSD mouse model. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay (ELISA), and immunofluorescence staining were used to evaluate the anti-inflammatory effects of XSG. The potential molecular mechanisms were explored and verified through network pharmacology analysis, Nissl staining, Western blot, ELISA, and RT-qPCR, respectively. Results: The body weight and behavior tests showed that MCAO combined with CUMS successfully established the PSD models. XSG alleviated neuronal damage, reduced the expressions of pro-apoptotic proteins Caspase-3 and B-cell lymphoma-2 (BCL-2)-associated X (BAX), and increased the expression of anti-apoptotic protein BCL-2 in PSD mice (P<0.05 or P<0.01). XSG inhibited microglial activation and the expressions of pro-inflammatory cytokines including tumor necrosis factor-α, interleukin (IL)-1 β, and IL-6 via the toll-like receptor 4/nuclear factor kappa-B signaling pathway in PSD mice (P<0.05 or P<0.01). Furthermore, XSG decreased the expression of indoleamine 2,3-dioxygenase1 (IDO1) and increased the concentration of 5-hydroxytryptamine in PSD mice (P<0.05 or P<0.01). Conclusion: XSG could reverse the anxiety/depressionlike behaviors and reduce the neuronal injury in the hippocampus and prefrontal cortex of PSD mice, which may be a potential therapeutic agent for PSD. [ABSTRACT FROM AUTHOR]

Background: Lovastatin, the main lipid-lowering component in red yeast rice, is a golden anti-lipid drug, but its long-term application is continuously challenged by potential skeletal muscle atrophy. Daidzein, an isoflavone derived from soybeans and many Chinese medicines, shows therapeutic potential in treating muscle-related diseases and metabolic disorders. However, whether daidzein can improve lovastatin-induced muscle atrophy and the specific mechanism needs to further study. Methods: Lovastatin-induced mice and zebrafish muscle atrophy models were used to validate the protective effect of daidzein in vivo. And the lovastatin-induced C2C12 myotube atrophy model was employed to validate the therapeutic efficacy and investigate the specific mechanism of daidzein in vitro. We combined specific siRNA targeting FOXO3a and AMPK-selective inhibitor, agonist to elucidate AMPK/FOXO3a-dependent muscle-protective mechanism of daidzein. The anti-atrophy effects of daidzein through blockage of abnormal activation of AMPK/FOXO3a was presented in Immunofluorescence, H&E staining, Western blot, qRT-PCR. Serum creatine kinase level was detected by ELISA and we used mouse muscle grip instrument to detect the strength of mouse muscles. Results: In this study, we demonstrated that daidzein could dose-dependently alleviate lovastatin-induced mice skeletal muscle atrophy, reduce serum creatine kinase, and improve muscle grip strength in mice. Mechanistically, daidzein inhibited lovastatin-induced FOXO3a phosphorylation caused by AMPK activation, thereby inhibiting FOXO3a nuclear translocation to restrain the expression of muscle-related proteins Atrogin-1 and MuRF-1. In C2C12 myotube, administration of AMPK-selective inhibitor Compound C recapitulated the therapeutic effects of daidzein against lovastatin-induced myotubes atrophy, while the anti-atrophy effects of daidzein were lost in the presence of AMPK-selective agonist MK-3903. In lovastatin-induced mice muscle atrophy models, Compound C elicited similar anti-atrophy effects as daidzein, but this effect was not potentiated when it was applied in combination with daidzein, suggesting that daidzein exerted therapeutic efficacy dependent on blockage of AMPK activity. Conclusions: Our study identified daidzein as an effective component that ameliorated lovastatin-induced skeletal muscle atrophy through blockage of abnormal activation of AMPK/FOXO3a and transcriptional activation of genes encoding downstream muscle-related proteins. Our results also highlighted the therapeutic potential by regulating the AMPK/FOXO3a axis in management of statin-induced myotoxicity. [ABSTRACT FROM AUTHOR]

Physiological changes of trophoblast cells are associated with gestational diabetes mellitus (GDM), and miR-184 involved in the development of GDM can be a potential therapeutic target. Asiaticoside (AS) has potential therapeutic effects on GDM, but its effect on miR-184 in HTR-8/Svneo cells is a relatively unworked area. The present study aimed to explore the effect of AS and miR-184 on physiological changes of HTR-8/Svneo cells.After the cytotoxicity assay of AS, HTR-8/Svneo cells were transfected with miR-184 mimics and/or treated with AS. The mRNA level of miR-184 in different groups was determined by real-time reverse transcription polymerase chain reaction, the cell viability was detected by cell counting kit-8 assay, the cell migration was measured by scratch test, the protein expressions of matrix metalloproteinase-2 (MMP2), MMP9, protein kinase B (AKT) and p-AKT were determined by western blot.80 μM AS with a treatment time of 48 h had no cytotoxicity and even promoted cell viability of HTR-8/Svneo cells. AS could significantly inhibit the mRNA level of miR-184 in HTR-8/Svneo cells (p<0.05). The overexpression of miR-184 significantly suppressed the cell viability, migration, and protein expressions of MMP2, MMP9 and p-AKT/AKT; however, AS was able to reverse the inhibition effect of miR-184 to increase the cell viability, migration and protein expressions of MMP2, MMP9 and p-AKT/AKT in HTR-8/Svneo cells (p<0.05).AS can reverse the inhibition effect of miR-184 on HTR-8/Svneo cells to facilitate cell proliferation, migration and AKT phosphorylation. [ABSTRACT FROM AUTHOR]

Objective: Geniposide (GP) provides myocardial cells with protection against pyroptosis-induced damage. However, the mechanisms governing GP's effect on the thioredoxin-interacting protein (TXNIP)/nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) signaling pathway remain unclear. This study aimed to explore how GP alleviates post-myocardial infarction (MI)-induced pyroptosis through regulation of the TXNIP/NLRP3 pathway. Material and Methods: In vivo studies: MI models were established, mouse body weight, heart rate, and blood glucose levels were monitored, and methods, such as cardiac ultrasound, hematoxylin–eosin staining, triphenyltetrazolium chloride staining, terminal deoxynucleotidyl transferase 2'-deoxyuridine 5'-triphosphate nick end labeling staining, quantitative polymerase chain reaction (qPCR), and Western blot (WB), were used to explore the effect of GP on myocardial cell pyroptosis. We explored the role of NLRP3 in GP's antimyocardial cell pyroptosis through qPCR, WB, immunofluorescence, enzyme-linked immunosorbent assay (ELISA), and other methods. In vitro studies: A chronic hypoxia (CH) cell model was established, and detection methods, such as cell counting kit-8 assay, transmission electron microscopy, ELISA, and immunological assays, were used to explore the effects of GP on CH myocardial cell pyroptosis and GP's inhibition of the TXNIP/NLRP3 signaling pathway to resist CH myocardial cell pyroptosis. Results: In vivo studies revealed that after the treatment with GP, the infarct area of mice's hearts significantly decreased, cardiac structure and function notably improved, fibroblast proliferation in cardiac tissues decreased significantly, and the pyroptosis level of myocardial cells decreased. GP treatment significantly downregulated the expression levels of type I collagen (Col I), Col III, TXNIP NLRP3, caspase-1, and gasdermin D N-terminal (GSDMD-N). The inhibition of NLRP3 also reduced the expressions of NLRP3, TXNIP, caspase-1, and GSDMD-N in the cardiac tissue, which is concomitant with a decline in reactive oxygen species (ROS) production. In addition, in vitro studies unveiled that GP effectively alleviated pyroptosis in CH myocardial cells, reducing pyroptosis rates, interleukin (IL)-1β, IL-18, lactate dehydrogenase, and creatine kinase-muscle/brain levels. This protective effect was achieved by inhibiting the TXNIP/NLRP3 signaling pathway. Conclusion: GP greatly diminishes the extent of infarcted myocardial tissue and mitigates pyroptosis, which improves cardiac structure and function through modulation of the TXNIP/NLRP3 pathway. Furthermore, the inhibition of NLRP3 lowers the expressions of factors associated with pyroptosis in the cardiac tissue and reduces ROS production. [ABSTRACT FROM AUTHOR]

Objective: Sepsis-induced acute kidney injury (AKI) is considered as a life-threatening complication of sepsis. The purpose of this study is to clarify the involvement of annexin A3 (ANXA3) in sepsis-related AKI. Material and Methods: Lipopolysaccharide (LPS) was used to establish a cell model based on HK2 cells. ANXA3 expression was quantified through quantitative real-time polymerase chain reaction. Cell proliferative capacities were assessed through 5-ethynyl-2'-deoxyuridine proliferation, cell counting kit-8, and colony formation experiments. Flow cytometry was utilized to analyze apoptotic cells. Inflammatory and oxidative stress indicators were measured by employing corresponding commercial assay kits. Endoplasmic reticulum (ER) stress markers were quantified through western blot analysis. Results: ANXA3 levels were significantly elevated in HK2 cells treated with LPS and in serum samples obtained from patients with AKI and sepsis (P < 0.001). LPS treatment exacerbated cellular damage, leading to increased ER and oxidative stresses, apoptosis, and inflammation, whereas knocking down ANXA3 significantly reversed these changes (P < 0.001). Conclusion: Interference with ANXA3 protected HK2 cells from LPS-induced cell injury through inhibiting inflammation, oxidative stress, apoptosis, and ER stress. [ABSTRACT FROM AUTHOR]

Objective: Nasopharyngeal carcinoma (NPC), a highly invasive form of head and neck cancer, carries a significant risk of distant metastasis. NPC is particularly prevalent in Asia and has a high incidence in southern China. Cisplatin-diamminedichloroplatinum (DDP), a chemotherapy agent, is commonly employed in NPC treatment. Despite DDP's efficacy, many patients eventually develop resistance to it over the course of their therapy, which significantly hinders treatment outcomes. Cancer-associated fibroblasts (CAFs) are key components of the tumor micro-environment and contribute to tumor progression and chemotherapy resistance. Exosomes secreted by CAFs serve as crucial mediators of intercellular communication and participate in modulating diverse biological processes. This study aimed to explore how exosomes derived from CAFs contribute to DDP resistance in NPC. Material and Methods: An in vitro coculture system was used to simulate the interaction between CAFs and NPC cells, and exosomes secreted by CAFs were isolated and characterized. The expression of autophagy hallmark proteins was detected by Western blot and quantitative real-time polymerase chain reaction. Autophagy intensity was quantified using monodansylcadaverine staining, and cell proliferation was assessed by colony formation assays and methylthiazolyldiphenyl–tetrazolium assays. NPC cells were treated with autophagy inducers (rapamycin), and the expression of Ras homologue enriched in brain (Rheb), mammalian target of rapamycin complex (mTORC1), and UNC51-like kinase was detected. Immunofluorescence was used to determine the cellular localization and expression intensity of mTORC1, and the effect on DDP sensitivity was evaluated through cell proliferation rates. In addition, the exosome-mediated resistance mechanism was further validated using an in vivo xenograft tumor model. Results: Coculture of CAFs with NPC cells significantly promoted the proliferation of NPC cells (P < 0.01), significantly elevated the IC50 value of DDP (P < 0.01), and elevated the resistance of NPC cells to DDP. CAF-derived exosomes elevated autophagy hallmark proteins light chain 3B-II, Beclin, and increased the autophagy intensity (P < 0.01). CAF-derived exosomes promoted autophagy by inhibiting mTORC1 (P < 0.01). In the in vitro model, exosomes promoted the growth of tumor tissues (P < 0.01), and the inhibition of exosome secretion reversed the promotion effect of autophagy (P < 0.01) and elevated the sensitivity of NPC cells to DDP. Conclusion: CAF-derived exosomes promote protective autophagy in NPC cells through the Rheb/mTOR axis, and result in DDP resistance in NPC. [ABSTRACT FROM AUTHOR]

Objective: Septic cardiomyopathy (SCM), as a complication of the septic process, severely affects the myocardial function of patients, but its pathogenesis remains unclear. The article aims to explore the mechanism of tripartite motif 10 (TRIM10) in rats with SCM and provide animal experimental basis for the treatment and prevention of SCM. Material and Methods: An SCM rat model was constructed by intraperitoneal injection of lipopolysaccharide (LPS). Sh-NC and sh-TRIM10 groups were injected with sh-NC and sh-TRIM10 in the tail vein for 3 consecutive days before SCM modeling. The expression of TRIM10 was detected by Western blot and reverse transcription–polymerase chain reaction analyses. Hematoxylin–eosin staining was performed to observe pathological changes in myocardium. Cardiomyocyte apoptosis was detected by flow cytometry. Serum levels of cardiac troponin I, myohemoglobin, creatine kinase-MB, interleukin-18 (IL-18), interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α), superoxide dismutase, and glutathione peroxidase (GSH-Px) were detected by enzyme-linked immunosorbent assay. Apoptosis-related proteins and toll-like receptor 4 (TLR4)/nuclear transcription factor-κB (NF-κB) pathway-related proteins were explored by Western blot assay. Results: TRIM10 expression increased in the LPS group (P < 0.0001). Myocardial tissue injury in SCM rats was improved after TRIM10 reduction compared with that in the LPS group. Knockdown of TRIM10 decreased the levels of MDA (P < 0.01), IL-18 (P < 0.0001), IL-1β (P < 0.0001), and TNF-α (P < 0.0001) and increased the contents of SOD (P < 0.001) and GSH-Px (P < 0.001) compared with those in the LPS group. TRIM10 reduced the apoptosis of H9C2 cells (P < 0.0001). After TRIM10 interference, the expression of p-P65/P65 (P < 0.0001) and TLR4 (P < 0.0001) was decreased. Conclusion: TRIM10 knockdown can reduce inflammation, oxidative stress, and apoptosis in SCM rats and has a protective effect on cardiomyocytes, which may be attributed to the regulation of the TLR4/NF-κB pathway. [ABSTRACT FROM AUTHOR]

Type 2 diabetes mellitus (T2DM), a metabolic disorder, has the hallmarks of persistent hyperglycemia, insulin resistance, and dyslipidemia. Protein‐tyrosine phosphatase 1B (PTP1B) was found to be overexpressed in many tissues in the case of T2DM and involved in the negative regulation of insulin signaling. So, PTP1B inhibition can act as a therapeutic target for T2DM. Numerous studies claimed the anti‐inflammatory, hypoglycemic, hepatoprotective, and hypolipidemic activities of Dodonaea viscosa. Previously, we generated the high‐fat diet (HFD)‐low dose streptozotocin (STZ)–induced diabetic male mice model and treated it with a PTP1B inhibitor (5, 7‐dihydroxy‐3, 6‐dimethoxy‐2‐ (4‐methoxy‐3‐ (3‐methyl‐2‐enyl) phenyl)‐4H‐chromen‐4‐one), isolated from Dodonaea viscosa. In the current study, we aimed to investigate the De novo lipogenesis, adipocyte differentiation, augmentation of lipoproteins clearance, fatty acid uptake, antilipolysis activity, and hepatic steatosis of PTP1B inhibition in adipose and liver tissues of the HFD‐STZ–induced diabetic mice model. We found the retrieval of normal morphology of adipocytes and hepatocytes in the compound‐treated group. The biochemical parameters showed the gradual reduction of LDL, VLDL, TC, and TG in the serum of the compound‐treated group. To further test our hypothesis, real‐time PCR was performed, and data revealed the reduction of PTP1B and other inflammatory markers in both tissues, showing enhanced expression of insulin signaling markers (INSR, IRS1, IRS2, and PI3K). Our compound upregulated the adipogenic (PPARγ), lipogenic (SREBP1c, FAS, ACC, and DGAT2), lipoprotein clearance (LPL, LDLR, and VLDLR), fatty acid uptake (CD36 and FATP1), and lipid droplet forming (FSP27 and perilipin‐1) markers expressions in adipocytes and downregulated in hepatocytes. Furthermore, we found elevated cholesterol efflux (in adipose and liver) and decreased lipolysis in adipocytes and elevated in hepatocytes. Hence, we can conclude that our compound protects the adipocytes from abrupt lipolysis and stimulates adipocyte differentiation. In addition, it plays a hepatic protective role by shifting clearance and uptake of lipoproteins and fatty acids to the peripheral tissues and retrieving the fatty liver condition. [ABSTRACT FROM AUTHOR]

Background: Despite declining COVID-19 incidence, healthcare workers (HCWs) still face an elevated risk of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection. We developed a diagnostic multivariate model to predict positive reverse transcription polymerase chain reaction (RT-PCR) results in HCWs with suspected SARS-CoV-2 infection. Methods: We conducted a cross-sectional study on episodes involving suspected SARS-CoV-2 symptoms or close contact among HCWs in Bogotá, Colombia. Potential predictors were chosen based on clinical relevance, expert knowledge, and literature review. Logistic regression was used, and the best model was selected by evaluating model fit with Akaike Information Criterion (AIC), deviance, and maximum likelihood. Results: The study included 2498 episodes occurring between March 6, 2020, to February 2, 2022. The selected variables were age, socioeconomic status, occupation, service, symptoms (fever, cough, fatigue/weakness, diarrhea, anosmia or dysgeusia), asthma, history of SARS-CoV-2, vaccination status, and population-level RT-PCR positivity. The model achieved an AUC of 0.79 (95% CI 0.77–0.81), with 93% specificity, 36% sensitivity, and satisfactory calibration. Conclusions: We present an innovative diagnostic prediction model that as a special feature includes a variable that represents SARS-CoV-2 epidemiological situation. Given its performance, we suggest using the model differently based on the level of viral circulation in the population. In low SARS-CoV-2 circulation periods, the model could serve as a replacement diagnostic test to classify HCWs as infected or not, potentially reducing the need for RT-PCR. Conversely, in high viral circulation periods, the model could be used as a triage test due to its high specificity. If the model predicts a high probability of a positive RT-PCR result, the HCW may be considered infected, and no further testing is performed. If the model indicates a low probability, the HCW should undergo a COVID-19 test. In resource-limited settings, this model can help prioritize testing and reduce expenses. [ABSTRACT FROM AUTHOR]

Objective: To compare the effects of titanium-prepared platelet-rich fibrin (T-PRF) and leukocyte platelet-rich fibrin (L-PRF) on osteoblasts. Methods: Venous blood samples were collected from ten volunteer patients to obtain T-PRF and L-PRF. The T-PRF group was labelled as Group T, the L-PRF group as Group L, and the control group, which includes only osteoblasts, was Group K. PRF samples were added to cultured osteoblast cells and cell proliferation was assessed using an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide) assay. The effect of different groups on osteoblast proliferation was observed for 72 h and the results were analysed statistically. Additionally, real-time polymerase chain reaction (RT-PCR) was conducted to evaluate gene expression levels of COL1A1, ALP, OCN, and RUNX2 within 48 h. Results: We found that the MTT results at 24 h were significantly lower than those at 48 and 72 h (p = 0.036 and p < 0.001, respectively). L-PRF levels showed an increase from 24 to 48 h followed by a decrease from 48 to 72 h. T-PRF levels were seen to increase at both the 24–48 h and 48–72-hour intervals. The changes in the COLA1, OCN, ALP, and RUNX2 genes at 24 h and 48 h did not significantly differ among the groups (p > 0.05). Conclusions: In this study, we investigated the effects of T-PRF and L-PRF on osteoblast proliferation over a 72-hour period. Both groups improved osteoblast proliferation, however T-PRF group showed a consistent increase in osteoblast proliferation up to 72 h, in contrast to the L-PRF group. No differences in gene expression were found. However, osteoblastic marker genes can be significantly expressed over longer time periods. Therefore, long-term studies are needed. [ABSTRACT FROM AUTHOR]

Porcine hemagglutinating encephalomyelitis virus (PHEV), porcine pseudorabies virus (PRV), and classical swine fever virus (CSFV) are currently prevalent worldwide and cause similar neurological symptoms in infected pigs. It is very important to establish a detection method that can rapidly and accurately detect and differentiate these three viruses. Targeting the PHEV N gene, PRV gB gene, and CSFV 5′ untranslated region (5′UTR), three pairs of specific primers and probes were designed, and a triplex crystal digital reverse transcription-PCR (cdRT-PCR) was developed to detect PHEV, PRV, and CSFV. The results indicated that this assay had high sensitivity, and the limitation of detection (LODs) for PHEV, PRV, and CSFV were 4.812, 4.047, and 5.243 copies/reaction, respectively, which was about 50 times higher than that of multiplex real-time quantitative RT-PCR (RT-qPCR). This assay showed good specificity, without cross-reaction with other important swine pathogens, i.e., FMDV, PRRSV, PEDV, SIV, TGEV, PoRV, and PCV2. This assay had high repeatability, with intra-assay coefficients of variation (CVs) of 0.73–1.87%, and inter-assay CVs of 0.57–2.95%. The developed assay was used to test 1,367 clinical tissue samples from Guangxi province in China, and the positive rates of PHEV, PRV, and CSFV were 3.44% (47/1,367), 1.24% (17/1,367), and 1.90% (26/1,367), respectively, with a coincidence rate of 98.98% and a Kappa value of 0.94 to the reference multiplex RT-qPCR. The established triplex cdRT-PCR was a highly rapid, sensitive, and accurate assay to detect and differentiate PHEV, PRV, and CSFV. [ABSTRACT FROM AUTHOR]

Background: A precise observation is that the cervix's solid tumors possess hypoxic regions where the oxygen concentration drops below 1.5%. Hypoxia negatively impacts the host's immune system and significantly diminishes the effectiveness of several treatments, including radiotherapy and chemotherapy. Utilizing oncolytic spores of Clostridium novyi-NT to target the hypoxic regions of solid tumors has emerged as a noteworthy treatment strategy. Methods: The transplantation procedure involved injecting TC-1 cells, capable of expressing HPV-16 E6/7 oncoproteins, into the subcutaneous layer of 6-8-week-old female C57/BL6 mice. The TC-1 cell line, was subcutaneously transplanted into 6-8-week-old female C57/BL6 mice. The tumor-bearing mice were randomly divided into 4 groups, and after selecting the control group, they were treated with different methods. Group 1- control without treatment (0.1 ml sterile PBS intratumor) Group 2- received cisplatin intraperitoneally (10 mg/kg) Group 3- received 107Clostridium novyi-NT spores systemically through the tail vein Group 4-tumor mice received 107Clostridium novyi-NT spores intratumorally. 20 days after the start of treatment, the mice were sacrificed and tumor tissues were isolated. In order to clarify the mechanism of the therapeutic effect with spores, the amount of ROS and ceramide was measured by ELISA technique, and the expression level of cytochrome c, cleaved caspase- 3, Bax, Bcl-2, HIF-1α, and VEGF proteins was measured by western blotting. Results: Our results clearly showed that the injection of Clostridium novyi-NT spores (either intratumorally or intravenously) causes the regression of mouse cervical tumors. Spore germination induces internal apoptosis in cancer cells by inducing ROS production and increasing total cell ceramide, releasing cytochrome c and damaging mitochondria. Additionally, the results provided clear evidence of a significant decrease in the expression of HIF-1 alpha and VEGF proteins among the tumor groups that received spores, when compared to both the cisplatin-treated group and the control group. Conclusions: The study's outcomes demonstrated that the introduction of Clostridium novyi-NT spores triggered apoptosis in cervical cancer cells (derived from the TC-1 cell line) via the mitochondrial pathway, subsequently resulting in tumor regression in a mouse model. [ABSTRACT FROM AUTHOR]

Background: Increased ribosome biogenesis is required for tumor growth. In this study, we investigated the function and underlying molecular mechanism of ribosome biogenesis factor (RBIS) in the progression of non-small cell lung cancer (NSCLC). Methods: In our study, we conducted a comprehensive analysis to identify key genes implicated in ribosome biogenesis by leveraging a Gene Set Enrichment Analysis (GSEA) dataset. Subsequently, we performed a comparative analysis of gene expression profiles by utilizing data from the Gene Expression Omnibus (GEO) datasets to ascertain differentially expressed genes (DEGs) between cancerous and adjacent non-cancerous tissues. Through the intersection of gene sets derived from GSEA and GEO, we identified a cohort of ribosome-associated genes that might exert a substantial influence on the progression of lung adenocarcinoma. Following an extensive literature review, we have identified the RBIS gene as an interesting candidate for further investigation. To elucidate the in vitro functional role of RBIS, several assays was employed, including the Transwell migration and invasion assay, wound healing assay, Cell Counting Kit-8 (CCK-8) proliferation assay, and colony formation assay. Subcutaneous and tail vein injection-based lung metastasis xenograft tumor models were used in evaluating the tumorigenic potential, growth, and metastatic spread of lung cancer cells. Flow cytometry analysis was employed to investigate cell cycle distribution and apoptotic rates. Additionally, real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was utilized to quantify the mRNA expression levels of genes. To comprehensively assess the translational efficiency of nascent proteins, we employed polysome profiling analysis to provide insights into the cellular translational landscape. Furthermore, we quantified global protein synthesis using a fluorescence-based assay to measure protein synthesis rates. The immunofluorescence technology was utilized to study the subcellular reorganization of the nucleolus. We conducted co-immunoprecipitation (Co-IP) assays followed by Western blot analysis to identify potential proteins interacted with RBIS. The half maximal inhibitory concentration (IC50) was used for evaluating the chemosensitivity of lung cancer cells to gemcitabine. Additionally, the colony formation assay was employed to assess the survival and proliferative capacity post-treatment of gemcitabine. Results: The database analysis showed that RBIS was upregulated in lung adenocarcinoma, and its high expression was associated with poor prognosis; Knockdown of RBIS significantly inhibited NSCLC cell migration, invasion and proliferation in vitro and xenograft tumor growth and metastasis in vivo. Additionally, knockdown of RBIS led to G0/G1 phase arrest and significantly increased apoptosis in lung adenocarcinoma cells. Mechanistically, downregulation of RBIS significantly decreased the expression of 47S ribosomal RNA (rRNA), a component associated with ribosome assembly. Polysome profiling analysis indicated that RBIS knockdown affected protein translation efficiency, and global protein synthesis assay further verified that RBIS knockdown inhibited synthesis of newborn proteins. Additionally, the ribosomal biogenesis-targeting drugs CX-5461 and the loss of RBIS exhibited synergistic effects in inhibiting cell cycle progression and inducing apoptosis. Furthermore, the ribosomal maturation factor GNL2 was identified as the key downstream regulator of RBIS in ribosome biogenesis. Notably, knockdown of RBIS substantially increased the sensitivity of lung adenocarcinoma cells to the chemotherapeutic drug gemcitabine, highlighting its l role in chemotherapy. Conclusions: Collectively, these studies suggested the close involvement of RBIS in the progression of lung adenocarcinoma, providing new insights for targeted therapeutic interventions involving ribosomes. [ABSTRACT FROM AUTHOR]

Summary Virus‐derived small interfering RNAs (vsiRNAs) play an important role in viral infection by regulating the expression of host genes. At present, research on the regulation of plant primary metabolic pathways by vsiRNAs is very limited. TvsiRNA24 derived from tobacco curly shoot virus (TbCSV) was amplified by reverse transcription polymerase chain reaction, and its target gene NbTPI (triosephosphate isomerase) was verified using reverse transcription quantitative polymerase chain reaction and GFP fluorescence observation. The effect of the interaction between TvsiRNA24 and NbTPI on TbCSV infection was analyzed by virus mediated, genetic transformation, western blotting, and quantitative detection. The expression of TvsiRNA24 retards the growth of Nicotiana benthamiana and enhances TbCSV accumulation within N. benthamiana. The overexpression of NbTPI attenuates the accumulation of TbCSV, and the silencing of NbTPI leads to the growth retardation of N. benthamiana and intensifies symptoms post‐TbCSV infection. Moreover, the expression of some genes related to photosynthesis, primary metabolism and immune response is regulated by NbTPI. Our results unveil the specific role of TvsiRNA24‐NbTPI in the pathogenicity of TbCSV, resulting in hindrance to plant growth and facilitation of viral infection. The identification of this regulatory pathway provides valuable insights that can be utilized to devise novel antiviral approaches targeting the reduction of viral pathogenicity. [ABSTRACT FROM AUTHOR]

ABSTRACT This study attempted to identify the relevant pathways involved in autophagy activation of pancreatic cancer and explore the mechanisms underlying immune evasion. Western blot (WB) was used to detect the expression of ITGB4, BNIP3, autophagy‐related proteins and MHC‐I. Co‐immunoprecipitation (Co‐IP) was used to verify the binding mode of ITGB4 and BNIP3. Flow cytometry was used to detect the expression of MHC‐I on the cell membrane. Transmission electron microscope (TEM) was used to observe cell autophagy. Confocal microscopy was used to observe the co‐localisation relationship between MHC‐I and autophagosomes in cells. ELISA was used to detect the level of lactate dehydrogenase and granzyme B in a tumour cell‐CD8+ T‐cell co‐culture system. Mouse syngeneic transplant tumour model and orthotopic tumour model were constructed and treated with PD‐1 monoclonal antibody to observe tumour growth. Quantitative reverse transcription polymerase chain reaction (qRT‐PCR) was used to detect the mRNA expression of ITGB4 and BNIP3 in tumour tissues. WB was used to determine the expression of autophagy‐related proteins. Flow cytometry was used to detect the expression of MHC‐I on cell membranes and the proportion of CD3+ and CD8+ cells. The results of Co‐IP experiments showed that ITGB4 could bind to BNIP3. It was observed under confocal microscopy that activating ITGB4/BNIP3 could promote the phagocytosis of MHC‐I by autophagosomes. Finally, the subcutaneous tumour transplantation and orthotopic tumour experiments in mice demonstrated the downregulation of ITGB4 significantly improved the therapeutic effect of PD‐1 antibodies on pancreatic cancer. In pancreatic cancer cells, autophagy is positively correlated with the ITGB4‐BNIP3 complex protein expression level. Autophagy diminishes the protein expression of MHC‐I, thereby promoting immune escape in pancreatic cancer cells. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSV) has emerged as a significant threat to the pig farming industry worldwide, resulting in considerable economic losses. However, few reports detail its economic impact on the pig farming sector. A study was conducted on 23 breeding pig farms in Hubei Province from January 2021 to December 2023, and the PRRSV infection status and associated economic losses were monitored to address this gap. PRRSV antigens and antibodies were identified through enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Additional monthly production data and weaning costs were gathered. The Kruskal-Wallis nonparametric test was used to assess the differences in production efficiency and weaning costs across various PRRSV infection statuses. Dunn's test was used to compare multiple groups. The parameter distributions of various variables were determined via @RISK (V. 8.5.2) software. Models were developed to evaluate the economic impact of PRRSV infection status on breeding pig farms and assess the losses from a PRRSV outbreak in either provisional PRRSV-negative or PRRSV-positive stable farms. A total of 754 months of monitoring was conducted across 23 breeding pig farms, which included 131 months (17.37%) classified as PRRSV provisional negative, 298 months (39.52%) as PRRSV-positive stable, and 325 months (43.11%) as PRRSV-positive unstable. The production efficiency and weaning costs were similar between provisional PRRSV-negative farms and PRRSV-positive stable farms, revealing no significant differences. However, these metrics varied significantly compared with those of PRRSV-positive unstable farms. With respect to provisional PRRSV-negative farms, PRRSV-positive stable farms and unstable farms faced additional annual losses of ¥3,135.17 and ¥4,898.79 per sow, respectively. Compared with PRRSV-positive stable farms, PRRSV-positive unstable farms incurred an extra annual loss of ¥1,763.62 per sow. Upon a PRRSV outbreak on a swine farm, followed by a return to preoutbreak conditions, the average economic loss per sow on provisional PRRSV-negative farms is approximately ¥3,061.21. Conversely, PRRSV-positive stable farms face an average loss of approximately ¥508.42 per sow. This study provides a systematic evaluation of the economic impact of PRRSV on Chinese pig farms, offering data to support the quantitative assessment of economic losses stemming from PRRSV within the domestic pig farming industry. [ABSTRACT FROM AUTHOR]