Melatonin Delays Arthritis Inflammation and Reduces Cartilage Matrix Degradation through the SIRT1-Mediated NF-κB/Nrf2/TGF-ß/BMPs Pathway.

Cartilage, a flexible and smooth connective tissue that envelops the surfaces of synovial joints, relies on chondrocytes for extracellular matrix (ECM) production and the maintenance of its structural and functional integrity. Melatonin (MT), renowned for its anti-inflammatory and antioxidant properties, holds the potential to modulate cartilage regeneration and degradation. Therefore, the present study was devoted to elucidating the mechanism of MT on chondrocytes. The in vivo experiment consisted of three groups: Sham (only the skin tissue was incised), Model (using the anterior cruciate ligament transection (ACLT) method), and MT (30 mg/kg), with sample extraction following 12 weeks of administration. Pathological alterations in articular cartilage, synovium, and subchondral bone were evaluated using Safranin O-fast green staining. Immunohistochemistry (ICH) analysis was employed to assess the expression of matrix degradation-related markers. The levels of serum cytokines were quantified via Enzyme-linked immunosorbent assay (ELISA) assays. In in vitro experiments, primary chondrocytes were divided into Control, Model, MT, negative control, and inhibitor groups. Western blotting (WB) and Quantitative RT-PCR (q-PCR) were used to detect Silent information regulator transcript-1 (SIRT1)/Nuclear factor kappa-B (NF-κB)/Nuclear factor erythroid-2-related factor 2 (Nrf2)/Transforming growth factor-beta (TGF-ß)/Bone morphogenetic proteins (BMPs)-related indicators. Immunofluorescence (IF) analysis was employed to examine the status of type II collagen (COL2A1), SIRT1, phosphorylated NF-κB p65 (p-p65), and phosphorylated mothers against decapentaplegic homolog 2 (p-Smad2). In vivo results revealed that the MT group exhibited a relatively smooth cartilage surface, modest chondrocyte loss, mild synovial hyperplasia, and increased subchondral bone thickness. ICH results showed that MT downregulated the expression of components related to matrix degradation. ELISA results showed that MT reduced serum inflammatory cytokine levels. In vitro experiments confirmed that MT upregulated the expression of SIRT1/Nrf2/TGF-ß/BMPs while inhibiting the NF-κB pathway and matrix degradation-related components. The introduction of the SIRT1 inhibitor Selisistat (EX527) reversed the effects of MT. Together, these findings suggest that MT has the potential to ameliorate inflammation, inhibit the release of matrix-degrading enzymes, and improve the cartilage condition. This study provides a new theoretical basis for understanding the role of MT in decelerating cartilage degradation and promoting chondrocyte repair in in vivo and in vitro cultured chondrocytes.

Nanomaterials for fluorescent assay of bilirubin.

The accumulation of bilirubin in blood is associated with many diseases. Sensitive and accurate detection of bilirubin is of great significance for personal health care. The rapid development of fluorescent nanomaterials promotes rapid development in the bilirubin assay. In this review, traditional methods for detection of bilirubin are briefly presented to compare with fluorescent nanosensors. Subsequently, the recent progress of different types of fluorescent nanomaterials for determination of bilirubin is summarized. Further, the performance of fluorescent nanosensors and conventional techniques for sensing bilirubin are compared. To this end, the challenges and prospects concerning the topics are discussed. This review will provide some introductory knowledge for researchers to understand the status and importance of fluorescent nanosensors for sensing bilirubin.

Facile one-pot synthesis of fluorescent aminoclay and the applications for fluorescence sensing Cr2 O7 2.

Here, we developed a facile one-pot strategy for the fabrication of fluorescent aminoclay (F-AC) through in situ solvothermal treatment of 3-aminopropyltrimethoxysilane, MgCl2 , and sodium ascorbate at 180°C for 6 h. The obtained F-AC exhibited blue emission, good water solubility, and satisfactory photostability. It was observed that Cr2 O7 2- could selectively quench the fluorescence of F-AC through the inner filter effect and static quenching process. As a result, a novel fluorescent F-AC-based nanosensor was constructed with good linearity in the range 0.1-75 µM. The nanosensor was successfully applied in real water samples with satisfactory results. This work not only provides a novel nanosensor for Cr2 O7 2- , but also highlights the F-AC's promising applications in wider fields due to the versatility and simplicity of the preparation strategy.

Association among biofilm formation, virulence gene expression, and antibiotic resistance in Proteus mirabilis isolates from diarrhetic animals in Northeast China.

BACKGROUND: The aim of this study was to investigate the association among biofilm formation, virulence gene expression, and antibiotic resistance in P. mirabilis isolates collected from diarrhetic animals (n = 176) in northeast China between September 2014 and October 2016. RESULTS: Approximately 92.05% of the isolates were biofilm producers, whereas 7.95% of the isolates were non-producers. The prevalence of virulence genes in the biofilm producer group was significantly higher than that in the non-producer group. Biofilm production was significantly associated with the expression of ureC, zapA, rsmA, hmpA, mrpA, atfA, and pmfA (P < 0.05). The results of drug susceptibility tests revealed that approximately 76.7% of the isolates were multidrug-resistant (MDR) and extensively drug-resistant (XDR). Biofilm production was significantly associated with resistance to doxycycline, tetracycline, sulfamethoxazole, kanamycin, and cephalothin (P < 0.05). Although the pathogenicity of the biofilm producers was stronger than that of the non-producers, the biofilm-forming ability of the isolates was not significantly associated with morbidity and mortality in mice (P > 0.05). CONCLUSION: Our findings suggested that a high level of multidrug resistance in P. mirabilis isolates obtained from diarrhetic animals in northeast China. The results of this study indicated that the positive rates of the genes expressed by biofilm-producing P. mirabilis isolates were significantly higher than those expressed by non-producing isolates.

Development and application of an indirect enzyme-linked immunosorbent assay based on recombinant capsid protein for the detection of mink circovirus infection.

BACKGROUND: Mink circovirus (MiCV) is a newly discovered pathogen associated with mink diarrhea. The prevalence and economic importance of this virus remain poorly understood, and no specific serological assay has been developed for the diagnosis of MiCV infection. RESULTS: In this study, a recombinant capsid protein antigen expressed in Escherichia coli was utilized to establish an indirect enzyme-linked immunosorbent assay (iELISA). Results revealed that the assay had no cross-reactivity with other related pathogens, and the respective sensitivity and specificity of the proposed iELISA were 92.31% and 91.67% compared with those obtained of Western blot on 138 serum samples from minks. The correlation coefficient between iELISA and Western blot was 0.838 (p > 0.05). iELISA was applied to detect MiCV antibodies in 683 clinical serum samples from different farms from the major mink industry province in China, and 21 of 24 farms with 163 of 683 (23.87%) individuals were tested positive for MiCV antibodies. The positive rates of each of the 21 flocks ranged from 2.33% to 73.68%. CONCLUSIONS: These results indicated that iELISA was a sensitive and specific method suitable for the large-scale detection of MiCV infections in mink. This study provided an effective method for the serological diagnosis and positive rate investigation of MiCV infection.

Unsupervised underwater shipwreck detection in side-scan sonar images based on domain-adaptive techniques.

Underwater object detection based on side-scan sonar (SSS) suffers from a lack of finely annotated data. This study aims to avoid the laborious task of annotation by achieving unsupervised underwater object detection through domain-adaptive object detection (DAOD). In DAOD, there exists a conflict between feature transferability and discriminability, suppressing the detection performance. To address this challenge, a domain collaborative bridging detector (DCBD) including intra-domain consistency constraint (IDCC) and domain collaborative bridging (DCB), is proposed. On one hand, previous static domain labels in adversarial-based methods hinder the domain discriminator from discerning subtle intra-domain discrepancies, thus decreasing feature transferability. IDCC addresses this by introducing contrastive learning to refine intra-domain similarity. On the other hand, DAOD encourages the feature extractor to extract domain-invariant features, overlooking potential discriminative signals embedded within domain attributes. DCB addresses this by complementing domain-invariant features with domain-relevant information, thereby bolstering feature discriminability. The feasibility of DCBD is validated using unlabeled underwater shipwrecks as a case study. Experiments show that our method achieves accuracy comparable to fully supervised methods in unsupervised SSS detection (92.16% AP50 and 98.50% recall), and achieves 52.6% AP50 on the famous benchmark dataset Foggy Cityscapes, exceeding the original state-of-the-art by 4.5%.

High-fat diet disrupts the gut microbiome, leading to inflammation, damage to tight junctions, and apoptosis and necrosis in Nyctereutes procyonoides intestines.

Given the burgeoning Nyctereutes procyonoides breeding industry and its growing scale, it is imperative to investigate the impact of high-fat diets on the health of these animals. This study involved 30 male Nyctereutes procyonoides of comparable weights (3 kg ±0.5), randomly assigned to either a control group or a high-fat diet group (n = 15 each). The latter group was fed a mixture of lard and basal diet in a 2:5 ratio, establishing a high-fat diet model in Nyctereutes procyonoides. This diet induced diarrhea and histopathological changes in the Nyctereutes procyonoides. Analysis of the small intestine contents using 16S rRNA sequencing revealed a high-fat diet-induced disruption in the gut microbiota. Specifically, Escherichia-Shigella emerged as the biomarker in the high-fat diet group (P = 0.049), while Vagococcus was prevalent in the control group (P = 0.049), indicating a significant increase in harmful bacteria in the high-fat diet group. Furthermore, this disrupted gut flora correlated with inflammation and oxidative stress, as evidenced by marked increases in TNF-α (P < 0.01), IL-1ß (P < 0.05), and IL-6 (P < 0.05) levels, measured via q-PCR, Western blot, and oxidative stress assays. In addition, q-PCR analysis revealed significant upregulation of apoptosis and necrosis markers, including Bax, Caspase3, Caspase9, Caspase12, RIPK3, and RIPK1 (P < 0.01 to P < 0.001), and a concurrent downregulation of the anti-apoptotic gene Bcl-2 (P < 0.01) in the high-fat diet group, consistent with protein expression trends. These findings suggest that a high-fat diet alters the gut microbiome toward a more harmful bacterial composition, escalating inflammatory responses and intestinal tissue permeability, culminating in intestinal cell apoptosis and necrosis.IMPORTANCEThis study examines the impact of high-fat diets on Nyctereutes procyonoides. Our research established a Nyctereutes procyonoides model on a high-fat diet, revealing significant health impacts, such as diarrhea, histological anomalies, and alterations in the gut microbiota. These findings emphasize the importance of preventing health issues and promoting sustainable industry growth. They highlight the significant impact of diet on gut microbiota and overall animal health.

Density functional theory study of neutral and oxidized thiophene oligomers.

The effect of oxidation on the energetics and structure of thiophene (Th) oligomers is studied with density functional theory at the B3PW91∕6-311++G(d,p) level. Neutral n-Th oligomers (2 < n < 13) are gently curved planar chains. Ionization potential and electron affinity results show that n-Th oligomers are easier to be oxidized as their chain length increases. Oxidation states +2, +4, +6, and +8 are energetically stable in 12-Th. Upon oxidation the conjugated backbone of 12-Th switches from extended benzenoid phase to quinoid phase localized on groups of monomers regularly spaced along the chain. Oxidized states +2, +4, +6, and +8 of 12-Th display two +1e localized at the ends of their chains only because of the finite size of the chains. In 12-Th this end-effect extends over the two terminal monomers forming a positive-negative charge duet. This peculiar charge localization makes n-Th oligomers different from other conducting polymers with similar structure, such as polypyrrole. The spectrum of single-electron molecular states of oxidized 12-Th displays two localized single-electron states in the HOMO-LUMO energy gap per +2 oxidation state. Oligothiophene 12-Th doped with F atoms at 1:2 concentration presents a charge transfer of 3.4 e from oligomer to dopants that increases to 4.8 e in the presence of solvent. The charge distribution in these F-doped oligomers is similar to the +4 oxidation state of 12-Th. It is predicted that dopants produce an enhanced charge transfer localized in the proximity of their locations enhancing the formation of bipolarons in the central part of the oligomer chain.

Carbon quantum dots for fluorescent detection of nitrite: A review.

NO2- is commonly found in foods and the environment, and excessive intake of NO2- poses serious hazards to human health. Thus, rapid and accurate assay of NO2- is of considerable significance. Traditional instrumental approaches for detection of NO2- faced with limitations of expensive instruments and complicated operations. Current gold standards for sensing NO2- are Griess assay and 2,3-diaminonaphthalene assay, which suffer from slow detection kinetics and bad water solubility. The newly emerged carbon quantum dots (CQDs) exhibit integrated merits including easy fabrication, low-cost, high quantum yield, excellent photostability, tunable emission behavior, good water solubility and low toxicity, which make CQDs be widely applied to fluorescent assay of NO2-. In this review, synthetic strategies of CQDs are briefly presented. Advances of CQDs for fluorescent detection of NO2- are systematically highlighted. Lastly, the challenges and perspectives in the field are discussed.

Oleanolic acid, a small-molecule natural product, inhibits ECM degeneration in osteoarthritis by regulating the Hippo/YAP and Wnt/ß-catenin pathways.

Over the past few decades, osteoarthritis (OA) has been a major health problem worldwide. It is urgent to develop new, effective, and safe drugs to treat OA. There are many pentacyclic triterpenoids in nature that are safe and have health benefits. Oleanolic acid (OLA), one of the pentacyclic triterpenoids, is a potential novel compound for treating OA; however, its mechanism of action is still unclear. In this study, the mechanism of resistance to extracellular matrix (ECM) degradation of OLA and its protective role in the amelioration of OA were investigated by in vivo and in vitro experiments. We found that OLA promoted interleukin-1ß (IL-1ß)-induced production of type II collagen (collagen II) in rat chondrocytes, decreased the expression of matrix metalloproteinase (MMP)-3 and MMP-13, and inhibited inflammatory cytokine (IL-1ß and TNF-α) and cartilage marker (CTX-II and COMP) levels, thereby hindering the pathological process of cartilage. Mechanistically, OLA inhibited the Wnt/ß-catenin pathway, activated the Hippo/YAP pathway, and hampered the ECM degradation process by inhibiting the nuclear translocation of ß-catenin and YAP. When we knocked down ß-catenin, OLA lost its stimulatory effect on the Hippo pathway. These findings confirm that OLA plays an anti-ECM degradation role by regulating the Wnt/ß-catenin and Hippo/YAP pathways. Overall, this study provides a theoretical basis for developing highly effective and low-toxic natural products for the prevention and treatment of OA.

Application of Real-Time Quantitative PCR to Detect Mink Circovirus in Naturally and Experimentally Infected Minks.

The mink circovirus (MiCV), a newly discovered pathogen, is associated with diarrhea in farmed minks. The prevalence and economic importance of this virus remain poorly understood, and a quantitative method for diagnosis of MiCV infection has not been established. This research aims to develop a highly specific, sensitive, and quantitative assay for MiCV. A Real-Time quantitative polymerase chain reaction (qPCR) assay was developed to detect different isolates of the MiCV in mink samples. The qPCR system is highly sensitive with a detection limit of as low as 10 viral DNA copies. The specificity of this qPCR assay was supported by the absence of cross-reaction with other pathogens. The coefficients of variation were low for both inter-assay and intra-assay variabilities. In addition, the results also expressed the distribution of MiCV in infectious mink tissues with high levels of virus in the skeletal muscle and heart. The heart occupied a higher proportion than other tissues, which can be considered the primary source of test material. This qPCR method could be a useful tool for epidemiological studies and disease management. This method for MiCV is highly specific, sensitive, repeatable, quantitative, and can rapidly determine viral load levels in different tissues samples.