Chloranthus holostegius, a perennial herbaceous plant of the family Chloranthaceae, is predominantly distributed in southern China and exhibits significant value in both horticultural and medicinal applications. In this study, nine previously undescribed lindenane-type sesquiterpenoid homo/heterodimers with diverse skeletons, designated as chlorholidenes A-I (1-9), including two novel naphthalene-fused norlindenane sesquiterpenoid dimers (1-2), two lindenane-type sesquiterpenoid dimers with naphthalene skeleton and 18-membered macrocycle (3-4), a 8,9-seco lindenane-type sesquiterpenoid dimer with an 18-membered macrocycle (5), a 2,3-seco lindenane-type dimer with [6 + 6] cycloaddition-formed 12-membered ring (6), an eudesmane-lindenane heterodimer (7), an lindenane-type sesquiterpenoid dimer with an oxaspiro [4.5]decane skeleton (8), and a classical lindenane dimer (9), together with two known acorane-lindenane heterodimers (10-11), were isolated from C. holostegius. Their structures were elucidated using 1D/2D NMR, HRESIMS, and ECD calculations. In vitro antiinflammatory assays showed compounds 3, 4, and 11 exhibited significant inhibition of LPS-induced NO release in RAW 264.7 cells (IC50: 12.14-16.13 & micro;M), outperforming dexamethasone (IC50 = 18.43 +/- 3.17 & micro;M). Compound 3 also suppressed neutrophil recruitment and ROS accumulation in CuSO4-induced inflammatory zebrafish in vivo. Mechanistically, compound 3 regulated the NF-kappa B/JAK pathways by inhibiting phosphorylation of key proteins (IKK-alpha/beta, I kappa B-alpha, p65, JAK1-3, TYK2) and blocking NF-kappa B p65 nuclear translocation. This study enriches lindenane sesquiterpenoid dimers structural diversity and identifies compound 3 as a promising lead for antiinflammatory drug development.

Five new sesquiterpenoids, named torijapolides A‒E, including three germacrane-type (1‒3) and two cadinene-type sesquiterpenoids (4‒5), were isolated from the fruits of Torilis japonica. Their structures were elucidated through comprehensive spectroscopic analysis, including MS, UV, IR, and 1D/2D NMR and quantum chemical calculations of electronic circular dichroism (ECD). In bioactivity evaluation, all isolates were evaluated for their inhibition effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced RAW 264.7 macrophage cells. Among them, compounds 1 and 2 exhibited significant inhibition of NO release, with IC50 values of 8.38 ± 0.06 µM and 10.27 ± 0.68 µM, respectively.

Endometrial cancer (EC) is a prevalent malignancy in women. UBE2T, a member of the E2 ubiquitin-conjugating enzyme family, has emerged as a potential regulator of cancer progression. In this study, we conducted WGCNA and machine learning analysis using the GEO dataset to identify UBE2T as a key target. Various experimental techniques, including qPCR, WB, IHC, CCK8, EdU, LDH release assays, MeRIP-qPCR and CHX experiments were employed to investigate the expression and function of UBE2T in EC. Additionally, we investigated the mechanism by which UBE2T is regulated through m6A modification. UBE2T is highly expressed in EC and exhibits low m6A modification levels. Knockdown of UBE2T significantly inhibits EC cell proliferation. Both FTO knockdown and METTL3 overexpression increase m6A modification of UBE2T and reduce its expression, respectively. Cycloleucine (CYC) treatment also promoted EC cell proliferation, and overexpression of FTO increased UBE2T expression and enhanced EC cell proliferation. Rescue experiments demonstrated that CYC treatment can reverse the increased m6A modification and restored UBE2T expression following FTO knockdown. Additionally, CYC treatment also rescues the proliferation inhibition caused by UBE2T knockdown in EC cells. Furthermore, UBE2T accelerates P53 protein degradation in EC cell lines. UBE2T is regulated by m6A modification and plays a crucial role in the progression of endometrial cancer by modulating key cellular processes. Targeting its expression or activity may offer a promising therapeutic strategy for intervention.

Osteoarthritis (OA) is a degenerative joint disease characterised by cartilage breakdown, leading to pain and reduced quality of life. This study aims to investigate the therapeutic potential of Puerarin (PUE), a natural compound derived from Pueraria lobata , in modulating OA progression. Employing a multifaceted approach that included bioinformatics analysis, molecular docking, in vitro assays, and in vivo experiments, we identified 57 overlapping targets between PUE and OA-related genes, suggesting a multi-target interaction model. Our findings revealed that PUE effectively inhibited inflammatory cytokine production and protected chondrocytes from apoptosis at non-cytotoxic concentrations (5 and 10 μM), promoting extracellular matrix synthesis by reversing IL-1β-induced degradation of Aggrecan and Collagen II while reducing MMP-13 and ADAMTS5 expression. Furthermore, PUE was shown to attenuate IL-1β-induced apoptosis by restoring BCL-2 levels and decreasing cleaved caspase-3 levels. In vivo, PUE administration in a destabilisation of the medial meniscus (DMM) mouse model significantly slowed OA progression, preserving cartilage structure and reducing osteophyte formation. Moreover, PUE activated the PI3K-AKT signalling pathway, underscoring its anti-inflammatory and anti-apoptotic mechanisms. Collectively, these results support Puerarin's potential as a disease-modifying agent in OA treatment, warranting further clinical exploration and consideration of its application in combination therapies to enhance cartilage protection and repair.

Pain hypersensitivity is a hallmark of rheumatoid arthritis (RA); however, the underlying mechanisms and effective therapies remain largely undefined. Emerging studies suggest that neutrophils play a significant role in the pathology of RA, yet their involvement in RA-associated pain is still unclear. The present study investigates whether neutrophil activity contributes to pain pathogenesis in RA. Our flow cytometry analysis reveals that the accumulation and N1 polarization (indicated by the ratio of CD45+CD66b+CD95+ subset) of neutrophils occur in synovial fluid samples from RA patients, positively correlating with pain scores. In the collagen-induced rheumatoid arthritis (CIA) model, mice demonstrate neutrophil accumulation, N1 polarization (indicated by the ratio of CD45+Ly-6G+CD95+ subset), and reactive oxygen species (ROS) production in affected paw tissues. Geranyl hydroquinone (GHQ), a natural meroterpenoid with antioxidative properties, reverses N1 polarization and ROS production in synovial neutrophils from RA patients in vitro. Moreover, a 10-day oral administration of GHQ alleviates pain hypersensitivity and reduces neutrophil accumulation, N1 polarization, and ROS production in CIA mice. Notably, GHQ treatment reverses TNF-α-evoked ROS production in neutrophils in vitro through downregulating gene expression associated with the ROS pathway. Further, liquid chromatography-tandem mass spectrometry and biochemical analyses indicate that GHQ binds to microsomal glutathione S-transferase 3 (MGST3) in neutrophils. In vitro and in vivo evidence demonstrates that the RA-specific analgesic and antioxidative effects of GHQ require MGST3. Lastly, GHQ administration exhibits superior therapeutic effects compared to methotrexate, a first-line disease-modifying antirheumatic drug, in CIA mice. Collectively, our findings indicate that neutrophil accumulation, N1 polarization and ROS production contribute to RA-associated pain, suggesting that targeting these pathways, such as with GHQ, could be a viable strategy for RA treatment.

OBJECTIVE:This study aimed to elucidate how DHA enhances the radiosensitivity of BC and to explain its potential mechanisms of action. METHODS:The circular structure of hsa_circ_0001610 was confirmed by Sanger sequencing, RNase R treatment, RT-PCR analysis using gDNA or cDNA. Cellular localization of hsa_circ_0001610 and microRNA-139-5p (miR-139-5p) was detected by fluorescence in situ hybridization. Cell counting kit-8 assay, wound healing and colony formation tests for assessing cell proliferation, while flow cytometry was utilized to estimate cell cycle progression and apoptosis. Reactive oxygen species and malondialdehyde experiments were conducted to validate ferroptosis of BC cells. The expression of ncRNAs and mRNAs was quantified via qRT-PCR, and protein expression was analyzed using Western blot. The effects of hsa_circ_0001610 and DHA on radiosensitivity of BC in vivo were studied by establishing BC mice model. RESULTS:In vivo and in vitro experimental results indicate that DHA promotes ferroptosis of BC cells at least partly by inhibiting hsa_circ_0001610/miR-139-5p/SLC7A11 pathway, thereby enhancing the radiosensitivity of BC cells. CONCLUSIONS:Our findings showed that DHA can induce ferroptosis of BC cells by down-regulation of hsa_circ_0001610, thus enhancing radiosensitivity, suggesting a promising therapeutic strategy for enhancing BC radiosensitivity that is worthy of further exploration.

Abstract Background: Cancer-induced bone paincaused by advanced tumor bone metastasis remains a clinical challenge, and the underlying mechanisms of BCP remain unknown. This study aimed to screen the expression profile of circular RNAs in a BCP rat model and provide a new theoretical basis for the role of circular RNA in the occurrence and development of BCP. Methods: We established a BCP rat model. The top four differentially expressed circRNAs (DECs) in the model were validated by agarose gel electrophoresis and Sanger sequencing between the BCP group and sham group. A circRNA-miRNA-mRNA network was constructed based on the interactions among circRNAs, microRNA (miRNA), and mRNA, which were predicted by TargetScan. mRNA and circRNA expression levels were detected by quantitative RT-PCR. In addition, Western Blot was performed to identify the protein levels of p-ERK, ERK, and Col8a1. Results: CircRNA parent genes were mainly enriched in MAPK and neurodevelopmental signalling pathways. CircAkt3 and circMap4k1 were significantly up-regulated in the BCP group. CircaAkt3 may increase p-ERK expression by upregulating Col8a1, which may further activate the MAPK pathway. Conclusions: The circAkt3 pathway may influence the development of bone cancer pain by activating the MAPK signaling pathway. This study provided important targets for the development of therapeutic strategy against BCP.

Circular ribonucleic acids (circRNAs) are a class of long non-coding RNA that were once regarded as non-functional transcription byproducts. However, recent studies suggested that circRNAs may exhibit important regulatory roles in many critical biological pathways and disease pathologies. These studies have identified significantly differential expression profiles of circRNAs upon changes in physiological and pathological conditions of eukaryotic cells. Importantly, a substantial number of studies have suggested that circRNAs may play critical roles in organ injuries. This review aims to provide a summary of recent studies on circRNAs in organ injuries with respect to (1) changes in circRNAs expression patterns, (2) main mechanism axi(e)s, (3) therapeutic implications and (4) future study prospective. With the increasing attention to this research area and the advancement in high-throughput nucleic acid sequencing techniques, our knowledge of circRNAs may bring fruitful outcomes from basic and clinical research.

Treatment of bone cancer pain (BCP) caused by bone metastasis in advanced cancers remains a challenge in clinical oncology, and the underlying mechanisms of BCP are poorly understood. This study aimed to investigate the pathogenic roles of circular RNAs (circRNAs) in regulating cancer cell proliferation and BCP development. Eight differentially expressed circRNAs in the rat spinal cord were validated by agarose gel electrophoresis and Sanger sequencing. Expression of circRNAs and mRNAs was detected by quantitative RT-PCR. MTS assay and flow cytometry were performed to analyze cell proliferation and apoptosis, respectively. Differentially expressed mRNA profiles were characterized by deep RNA sequencing, hierarchical clustering, and functional categorization. The interactions among circRNAs, microRNAs (miRNAs), and mRNAs were predicted using TargetScan. Additionally, western blot was performed to determine the protein levels of Pax8, Isg15, and Cxcl10. Multiple circRNAs were differentially expressed in the spinal cords of BCP model rats; of these, circSlc7a11 showed the greatest increase in expression. The overexpression of circSlc7a11 significantly promoted cell proliferation and repressed apoptosis of LLC-WRC 256 and UMR-106 cells, whereas circSlc7a11 silencing produced the opposite effects. Altered expression of circSlc7a11 also induced substantial changes in the mRNA expression profiles of LLC-WRC 256 cells; these changes were linked to multiple apoptotic processes and signaling pathways, such as the chemokine signaling pathway, and formed a complex circRNA/miRNA/mRNA network. Additionally, Pax8, Isg15, and Cxc110 protein level in LLC-WRC 256 cells was consistent with the mRNA results. The circRNA circSlc7a11 regulates rat BCP development by modulating LLC-WRC 256 cell proliferation and apoptosis through multiple-signaling mechanisms.

The aim of this study was to explore the neurocognitive effects of dexmedetomidine-loaded gold nanoparticles (AuNPs-dexmedetomidine) on anesthetized rats. Sixty Sprague Dawley rats (age, 2-3 weeks; weight, 250-280 g) were randomly divided into three groups (n = 20): the control group and two groups that received intraperitoneal injection of AuNPs-dexmedetomidine at 50 and 100 μg/kg each. Western blotting and RT-PCR were used to determine the protein and mRNA expression of GSK-3β, respectively. Compared with that in the control group, GSK-3β expression in AuNP-dexmedetomidine groups increased (P < 0.05). The protein expression of GSK-3β was higher and mRNA expression was significantly lower in the 100 μg/kg AuNP-dexmedetomidine group (P < 0.05). AuNPs-dexmedetomidine reduced the neurocognitive effect on anesthetized rats through the regulation of the GSK-3β signaling pathway.

The original version of this article unfortunately contained a mistake in Fig. 2.

The authors have retracted this article [1] because in further experiments, they found that some experimental data cannot be verified repeatedly.

The original version of this article unfortunately contained a mistake in Fig. 2.

Bone cancer pain (BCP) is a common chronic pain that is caused by a primary or metastatic bone tumor. More detailed molecular mechanisms of BCP are warranted. In this study, we established a BCP rat model. The von Frey hair test, body weight, and hematoxylin and eosin staining were employed. We screened differentially expressed circRNAs (DECs) between the BCP group and sham group. The results revealed that 850 DECs were significantly up-regulated and 644 DECs were significantly down-regulated in the BCP group. Furthermore, we identified 1177 differentially expressed genes (DEGs) significantly up-regulated and 565 DEGs significantly down-regulated in the BCP group. Gene Ontology annotation of all 1742 DEGs revealed that biological regulation of metabolic processes, cellular processes, and binding were the top enriched terms. For Kyoto Encyclopedia of Genes and Genomes analysis, phagosome, HTLV-I infection, proteoglycans in cancer, and herpes simplex infection were significantly enriched in this study. In addition, we identified four selected circRNAs, chr6:72418120|72430205, chr20:7561057|7573740, chr18:69943105|69944476, and chr5:167516581|167558250, by quantitative real time PCR. chr6:72418120|72430205 (circStrn3) was selected for further study based on expression level and the circRNA-miRNA-mRNA network table. Western blot analysis suggested that knockdown of circStrn3 could effectively induce Walker 256 cell apoptosis. In summary, our study provided a more in-depth understanding of the molecular mechanisms of BCP.

Background: Bone cancer pain (BCP) is a common symptom occurring among patients with cancer and has a detrimental effect on their quality of life. Growing evidence has implicated microRNA-329 (miR-329) in the progression of bone diseases. In the present study, we aimed to elucidate the potential effects of miR-329 on BCP in a BCP mouse model via binding to lysophosphatidic acid receptor 1 (LPAR1) through the LPAR1/extracellular signal-regulated kinase (ERK) signaling pathway. Methods: Initially, a BCP mouse model was established via injection of 4 × 10 4 murine breast tumor (4T1 cell) cells (4 μl). The interaction between miR-329 and LPAR1 was identified using a bioinformatics website and dual luciferase reporter gene assay. The modeled mice were subsequently treated with miR-329 mimic, LPAR1 shRNA, or both, in order to examine the effect of miR-329 on the paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) of mice, the expression of LPAR1/ERK signaling pathway-related genes. Results: The positive expression rate of LPAR1 protein and extent of ERK1/2 phosphorylation were increased in BCP mouse models. LPAR1 is a target gene of miR-329, which can inhibit the expression of LPAR1. In response to miR-329 overexpression and LPAR1 silencing, BCP mice showed increased PWT and PWL, along with decreased LPAR1 expression and ratio of p-ERK/ERK. Conclusions: Altogether, the results obtained indicated that miR-329 can potentially alleviate BCP in mice via the inhibition of LPAR1 and blockade of the LPAR1/ERK signaling pathway, highlighting that upregulation of miR-329 could serve as a therapeutic target for BCP treatment.

Morphine is widely used in clinical practice for a class of analgesic drugs, long-term use of morphine will cause the action of tolerance. MicroRNAs have been reported to be involved in morphine analgesic tolerance..