Intervertebral disc degeneration (IVDD) is characterized by progressive nucleus pulposus cell loss and extracellular matrix degradation, in which persistent oxidative stress plays a critical pathogenic role. Transplantation of nucleus pulposus–derived stem cells (NPSCs) is a promising therapeutic strategy, yet the hostile oxidative microenvironment severely compromises cell survival. Although cellular quiescence has been suggested to enhance stress tolerance, its regulatory mechanisms and relevance in NPSCs remain largely unexplored. Oxidative stress was evaluated in human degenerated disc tissues, a rat needle-puncture degeneration model, and tert-butyl hydroperoxide–treated NPSCs in vitro. Proliferating and quiescent NPSCs were compared for reactive oxygen species (ROS) levels, apoptosis, viability, and transcriptomic profiles. Pathway enrichment analyses were performed to identify critical signaling mechanisms. Recombinant transforming growth factor-beta 3 (rhTGF-β3) was used to activate the pathway, while small interfering RNA targeting the transforming growth factor-beta receptor type 2 (Tgfbr2) and the pharmacological inhibitor SB431542 were applied for pathway suppression. Functional assays, organ culture, and in vivo transplantation were conducted to assess cell survival and regenerative effects. Elevated oxidative stress was consistently observed across clinical, animal, and cellular models of disc degeneration. Quiescent NPSCs demonstrated enhanced resistance to oxidative injury, with reduced ROS accumulation, decreased apoptosis, and improved survival. Transcriptomic analyses revealed suppression of metabolic and P53-mediated apoptotic pathways, alongside marked activation of TGF-β/SMAD signaling. Activation of this pathway induced quiescence, reduced ROS levels, inhibited mitochondrial apoptotic signaling, and protected NPSCs from oxidative injury, whereas pathway inhibition abolished these protective effects. In both organ culture and in vivo transplantation models, quiescent and TGF-β–activated NPSCs exhibited superior survival and significantly improved disc structural preservation compared with proliferating or pathway-blocked cells. Activation of the TGF-β/SMAD pathway induces NPSC quiescence and enhances oxidative stress tolerance by suppressing P53-dependent mitochondrial apoptosis. Pharmacological induction of quiescence represents a potential strategy to improve stem cell–based therapies for intervertebral disc degeneration.

Non-alcoholic fatty liver disease (NAFLD) is the leading cause of chronic liver diseases worldwide, necessitating urgent novel oral treatments. In this study, beta-cell expansion factor A (BefA) was evaluated in a murine NAFLD model induced by high-fat diet (HFD). Our results revealed that BefA significantly reduced body weight (36.58 f 1.55 g vs. 42.30 f 1.96 g, p < 0.01), fat mass-to-body weight ratio (0.023 f 0.019 vs. 0.300 f 0.019, p < 0.05), liver weight (1.90 f 0.07 g vs. 2.31 f 0.21 g, p < 0.05), and liver function parameters (ALT, AST, ALP levels reduced, p < 0.05). Notably, BefA reversed the pathological features of NAFLD, decreasing hepatic steatosis score from 3.67 f 0.47 to 1.67 f 0.47 (p < 0.01). Mechanistically, BefA activated the AMPK signaling pathway, resulting in the suppression of lipogenic gene transcription (ACC, FASN, SREBP-1c) and the enhancement of fatty acid oxidation (CPT-1, PPAR-alpha). However, AMPK inhibitor and broad-spectrum antibiotics significantly attenuated the benefits observed with BefA treatment, increasing body weight, fat-to-body weight ratio, and liver weight (p < 0.05). Similar detrimental effects were also observed in liver function indices and histopathological characteristics. These findings underscore the pivotal role of both gut microbiota modulation and AMPK signaling in BefA's therapeutic efficacy, making it a promising multitargeted approach for NAFLD treatment.

Background: Osteoarthritis (OA) is a leading cause of pain, disability, and reduced mobility worldwide, characterized by metabolic imbalances in chondrocytes, extracellular matrix (ECM), and subchondral bone. Emerging evidence highlights the critical role of long non-coding RNAs (lncRNAs) in OA pathogenesis. This study focuses on lncRNA PTS-1, a novel lncRNA, to explore its function and regulatory mechanisms in OA progression. Methods: The expression profile of lncRNAs was assessed using RNA sequencing and qRT-PCR. The expression of lnc-PTS-1 was further validated by qRT-PCR in degenerated cartilage tissues, degenerative primary chondrocytes, and IL-1(3-treated C28/I2 cells. Cell viability, proliferation, and apoptosis rates, along with the mRNA and protein levels of apoptosis-related markers (cleaved Caspase 3, cleaved Caspase 9, Bcl-2, Bax), ECM metabolism markers (MMP-3, MMP-13, aggrecan, collagen II), and inflammation-related markers (IL-1(3, IL-6, TNF-alpha) were evaluated using Cell Counting Kit-8, Toluidine Blue staining, Alcian Blue staining, flow cytometry, qRT-PCR, immunofluorescence, and Western Blot. The interaction between miR-8085 and lnc-PTS-1 or E2F2 was investigated through dual luciferase reporter assays and RNA immunoprecipitation (RIP) analyses. Results: Lnc-PTS-1 expression was significantly downregulated in degenerated cartilage tissues, IL-1(3-induced degenerative primary chondrocytes and C28/I2 cells. Functional experiments showed that lnc-PTS-1 knockdown aggravated IL-1(3-induced ECM degradation, chondrocyte apoptosis, and inflammation, while its overexpression provided protective effects. Mechanistically, lnc-PTS-1 acted as a competing endogenous RNA (ceRNA) by sponging miR-8085, thereby upregulating E2F2 expression. Notably, miR-8085 upregulation diminished the protective effects of lnc-PTS-1 on ECM degradation, apoptosis, and inflammation, while E2F2 upregulation partially alleviated IL-1(3-induced damage. However, these mitigating effects were reversed by miR-8085 overexpression. Conclusion: These findings identify lnc-PTS-1/miR-8085/E2F2 axis as a novel regulatory mechanism in OA pathogenesis, providing theoretical basis and experimental evidence for the potential clinical application of new lncRNA molecules in the treatment of OA.

Chronic cerebral hypoperfusion (CCH) is a major contributor to vascular dementia, with neuroinflammation playing a central role in its pathogenesis. Sinomenine (SINO), a natural alkaloid derived from traditional Chinese medicine, has shown significant anti-inflammatory and neuroprotective properties. However, its efficacy and mechanism of action in CCH remain unclear. In this study, we established a CCH rat model through bilateral common carotid artery occlusion and administered 10 mg/kg of SINO daily. Behavioral tests demonstrated that SINO significantly improved cognitive and memory functions in CCH rats. Histological analysis revealed that SINO effectively reduced neuroinflammation and damage in the hippocampal CA1, CA3, and DG regions. Mechanistically, SINO promoted microglial polarization from the M1 to M2 phenotype, markedly inhibiting the release of pro-inflammatory cytokines, including IL-1β, IL-6, and TNF-α. Further exploration of its neuroprotective mechanism showed that exosomes from SINO-treated microglia were enriched with miRNA-223-3p, which suppressed NLRP3-mediated pyroptosis in neurons. While our findings highlight the therapeutic potential of SINO, further studies are needed to validate its safety and efficacy in diverse populations and chronic settings. In summary, this study not only demonstrates SINO’s regulatory effect on microglial polarization in CCH but also unveils a novel neuroprotective mechanism through exosomal miRNA-223-3p delivery, providing a solid theoretical foundation for SINO’s potential as a treatment for CCH.

BackgroundOsteosarcoma, an aggressive bone malignancy predominantly affecting children and adolescents, presents significant therapeutic challenges with a 5-year survival rate below 30% in metastatic cases. T-cell exhaustion, characterized by the overexpression of immune checkpoint molecules, contributes to osteosarcoma progression and immune evasion. Although targeting these inhibitory pathways has shown potential in restoring T-cell activity, the molecular regulators of T-cell depletion in osteosarcoma are poorly understood.MethodsThis study employed comprehensive bioinformatics analyses on osteosarcoma samples from the TARGET database, combined with normal tissue data from the GTEx database, to identify T-cell exhaustion-associated genes and their co-expressed long non-coding RNAs (lncRNAs). Gene ontology and KEGG pathway analyses were used to elucidate immune-related pathway enrichments. A six-lncRNA prognostic model was established using LASSO regression and validated in separate cohorts. Functional assays evaluated the impact of the lncRNA AL031775.1 on osteosarcoma cell behavior and T-cell function.ResultsTwenty-four key T-cell exhaustion-related genes were identified and significantly enriched in immune-related pathways, indicating their importance in the osteosarcoma immune microenvironment. The constructed six-lncRNA model stratified patients by survival prognosis, showing robust predictive performance across cohorts. Among the six identified lncRNAs, AL031775.1 is notably downregulated in osteosarcoma patients and significantly promotes osteosarcoma cell proliferation, migration, and invasion while contributing to T-cell exhaustion. In T cells, downregulation of AL031775.1 impairs antitumor immunity, upregulates immune checkpoint molecules LAG3, PD1, and CTLA4, and diminishes T-cell cytotoxic activity against tumor cells.ConclusionThis study identifies a novel six-lncRNA prognostic model and highlights the therapeutic potential of AL031775.1 in managing osteosarcoma by enhancing T-cell immunity and counteracting tumor progression. Targeting AL031775.1 represents a promising approach to improve immunotherapy efficacy in osteosarcoma. These findings provide critical insights into the molecular regulation of T-cell exhaustion and suggest a new avenue for therapeutic intervention.

Osteosarcoma (OS) is a prevalent form of bone cancer among younger people, particularly children and adolescents. Ferroptosis is a non-apoptotic cell death identified by increased levels of iron-dependent lipid peroxidation. This study was designed to develop a prognostic model based on differentially expressed genes (DEGs) associated with ferroptosis and examined the functions of ferroptosis-related genes (FRGs) in OS cells. Gene expression profiles in OS were retrieved from TARGET and GEO databases, while GTEx provided data for healthy tissues. Prognostic genes were identified through bioinformatics analysis and data integration. In vitro experiments, cell cultures, qRT-PCR, immunohistochemistry (IHC), cell transfection, Edu assays, DHE assays, migration, and invasion assays validated the prognostic model and explored the functional role of FRGs in OS cells. Univariate Cox regression analysis demonstrated that 12 DEGs were differentially expressed. Based on four FRGs in OS constructed a risk-scoring model. The high-risk (HR) group showed a considerably lower OS rate than the low-risk (LR) group (p < 0.001 in the TARGET and p < 0.05 in the GSE21257 cohorts). A risk score was validated as an independent predictive factor for OS via multivariate Cox regression. Functional analysis shows that these FRGs affect the occurrence of ferroptosis by influencing the intracellular ROS levels and play a regulatory role in the proliferation, migration, and infiltration of OS cells. The findings suggested that four FRGs demonstrate significant prognostic value in OS, offering potential insights into novel therapeutic targets for OS treatment.

This study aimed to evaluate the effects of MAT2A silencing on the proliferation, migration, and invasion of gastric cancer (GC) cells and to investigate the underlying molecular mechanisms. MAT2A expression in GC tissues and adjacent normal tissues was assessed using immunohistochemistry and western blotting. Two GC cell lines with elevated MAT2A expression underwent gene silencing. Cellular proliferation, migration, and invasion were evaluated using EdU incorporation, Transwell, wound healing, and flow cytometry assays. Transcriptomic profiling was conducted to identify downstream pathways affected by MAT2A silencing, demonstrating significant enrichment of the p53 signaling pathway. To further clarify this mechanism, the p53 pathway was inhibited in MAT2A-silenced cells, and changes in p53 and p21 protein expression, along with alterations in proliferation, migration, and invasion, were reassessed. MAT2A expression was significantly higher in GC tissues and cell lines compared with adjacent normal controls. Silencing of MAT2A induced cell cycle arrest, suppressed proliferation and metastatic capacity, and enhanced apoptosis, accompanied by an increased expression of p53 and p21 proteins. Inhibition of p53 with PFT-α reduced both p53 and p21 levels and reversed the suppressive effects of MAT2A silencing on GC cell proliferation, migration, and invasion. MAT2A silencing induces cell cycle arrest, enhances apoptosis, and inhibits malignant phenotypes of GC cells, including proliferation, migration, and invasion, through activation of the p53 signaling pathway.

Idiopathic short stature (ISS) remains a major pediatric challenge with unclear causes and inconsistent responses to growth hormone therapy. Here we show that plasma exosomes from children with ISS contain elevated hsa-miR-17-3p that disrupts growth signaling and impairs cartilage cell proliferation. Elevated miR-17-3p suppresses ZNF148/SOS1 signaling, linking molecular dysfunction to dietary exposure in ISS. To investigate environmental triggers, we developed a capsaicin-rich diet rat model that recapitulates ISS, showing normal Gh/Igf-1 levels but elevated plasma miR-17-3p. The diet induced mild gut inflammation, increasing miR-17-3p in intestinal and plasma exosomes. Fecal samples from ISS children exhibited similar elevations in miR-17-3p and inflammatory markers, linking spicy diets to ISS pathogenesis. Finally, engineered exosomes designed to silence miR-17-3p, combined with localized growth hormone therapy, restored growth plate function. These findings uncover a diet-driven exosome axis underlying ISS and suggest new therapeutic strategies for children in high-capsaicin regions.

Dendritic cells (DCs) are crucial for the initiation and regulation of innate and adaptive immunity. Their maturity and infiltration in the tumor largely determine the efficiency of antigen presentation, the CTL responses, and the prognosis of tumors. However, the application of common immunoregulatory plant polysaccharides to DCs in vivo still represents major challenges due to the off-target effect and short biological lifespan. Lonicera japonica Thunb. polysaccharides (LJP) were found to exert benign immunoregulatory ability, but the effectiveness of utilizing LJP alone is unsatisfactory. As a result, we innovatively encapsulated LJP in into the exosomes derived from mouse bone mesenchymal stem cells (BMSCs) to form a DC-activated inducer (LJP-exosome). LJP-exosomes possessed a profound ability to target lymph nodes and the co-stimulatory capability of DCs compared with the application of LJP alone. Adequate results have shown that DCs primed by LJP-exosomes enhanced the tumor-reactive CD8+ T cell responses, leading to prophylactic tumor inhibition in an immunologically ignorant tumor model. The study proposed offers a promising strategy for enhancing the immune activation efficacy of extracted polysaccharides of traditional Chinese medicine by building the patients’ immunity, thus consolidating the overall prognosis.

Osteosarcoma, the most common primary bone cancer in children and adolescents, presents significant challenges, particularly in metastasis and recurrence, resulting in poor survival rates. This study explores the role of PANoptosis—a complex cell death mechanism involving pyroptosis, apoptosis, and necroptosis—in osteosarcoma by identifying relevant long non-coding RNAs (lncRNAs) and their prognostic significance. Bioinformatics analyses used RNA expression data from the GEO and TARGET databases to identify differentially expressed genes (DEGs) and PANoptosis-related genes (PRGs). Co-expression analysis revealed lncRNAs linked to PRGs, forming a risk prognostic model. Five PRGs and two lncRNAs were significantly associated with prognosis, with the model showing high predictive accuracy (AUC 0.876, 0.787, and 0.794 for 1, 3, and 5 years). Notably, lncRNA AC133552.2 was downregulated in osteosarcoma tissues, correlating with poor survival and reduced immune infiltration. Silencing AC133552.2 promoted cell proliferation and migration, while overexpression inhibited tumor growth and metastasis, confirmed in xenograft models. AC133552.2 emerges as a potential biomarker and therapeutic target, with future research needed to explore its molecular mechanisms and clinical application.

The objective of this study is to observe the antitumor efficacy of the second mitochondria-derived activator of caspases (SMAC) mimetic bivalent smac mimetic (BV6) in combination with target of rapamycin (mTOR) inhibitor on DDP (cisplatin) sensitivity. Ovarian cancer cells were exposed to cisplatin, BV6, DDP + BV6, and DDP + BV6 + mTOR inhibitor Rapamycin. Using proteomics and bioinformatics, protein expression profiles in ovarian cancer were determined. Bagg Albino color nude mice were treated with DDP or BV6 alone or in combination, or BV6 + DDP + Rapamycin. The effects of different treatments on ovarian cancer cells and tumor growth were evaluated in vivo and in vitro. Proteomics and bioinformatics analysis revealed significant changes of protein kinase (AKT)/mTOR pathway. Consistently, western blot data indicated that AKT/mTOR axis was gradually activated in BV6-treated ovarian cancer cells and attenuated the cytotoxic effect of BV6. Functional assays showed that DDP or BV6 treatment alone significantly enhanced the sensitivity and inhibited the migration of ovarian cancer cells, but without any synergistic effects. In addition, combination with BV6 and mTOR inhibitor Rapamycin significantly decreased cell viability and inhibited migration of ovarian cancer cells exposed to DDP. Consistently, the xenograft model showed that co-treatment with Rapamycin with BV6 had significantly suppressed tumor growth and metastasis. Our study demonstrated that SMAC analogue BV6 exhibits a strong anticancer effect on ovarian cancer in vitro and in vivo. Combination with Rapamycin overcomes the activation of mTOR pathway by BV6 and increases the chemosensitivity to DDP. These data suggest a potential application of triple combination with DDP + BV6 + Rapamycin in clinical management of ovarian cancer.

The early diagnosis of endometriosis is delayed and the clinical treatment is difficult, which causes severe economic burden to the patients. A comprehensive study of the cell classification and composition of endometriosis, is essential for early diagnosis and pathogenesis. This study utilized single-cell data as a reference to accurately deconvolute cell types within bulk transcriptomics data. Five main cell types in endometriosis (epithelial cells, mesenchymal cells, endothelial cells, lymphocytes, and myeloid cells) revealed varying degrees of change compared to healthy controls. Notably, epithelial cells significantly decreased while mesenchymal cells increased. However, MUC5B epithelial cells, showed an increasing trend, consistent with the increasing trend of dStromal-late mesenchymal cells and eM2 macrophages.Pathway enrichment analysis revealed the EMT signalling pathway dominated in endometriosis. Additionally, a random forest method based on cell types could successfully distinguish endometriosis patients from healthy control, which illustrates the potential value of cell types in early diagnosis.

BACKGROUND:Diabetic cardiomyopathy (DCM) stands as the primary cause of heart failure and mortality among patients with diabetes. Nevertheless, conventional treatment approaches are limited in their ability to effectively prevent myocardial tissue damage itself. Mesenchymal stem cell (MSC) therapy exhibits immense potential for treating DCM; however, the precise mechanisms involved in regulating inflammatory responses and pyroptosis processes, an emerging form of cellular death, within myocardial cells remain elusive. Hence, it is imperative to further elucidate the precise underlying mechanisms to facilitate the clinical implementation of MSC therapy. METHODS:In vivo, we established a DCM mouse model by administering streptozotocin and fed the mice a high-glucose and high-fat diet, followed by MSC therapy. Cardiac function and myocardial injury were evaluated through echocardiography and histological analysis. Furthermore, the levels of inflammation and pyroptosis were assessed using ELISA, Western blotting, and qRT-PCR. In vitro experiments involved inducing H9C2 myocardial cell damage with high glucose treatment, followed by coculture with MSCs to investigate their role in modulating inflammation and pyroptosis mechanisms. RESULTS:MSCs can maintain cardiac function and alleviate myocardial injury in mice with DCM. Moreover, they effectively suppress the activation of NLRP3 and reduce the release of inflammatory factors (such as IL-1β and ROS), thereby further downregulating the expression of pyroptosis-related proteins including NLRP3, Caspase-1, and GSDMD. Additionally, we experimentally validated that MSCs exert their therapeutic effects by promoting the expression of miR-223-3p in cardiac myocytes; however, this effect can be reversed by an miR-223-3p inhibitor. CONCLUSION:MSCs effectively mitigate the release of inflammatory factors and cell lysis caused by pyroptosis through the regulation of the miR-223-3p/NLRP3 pathway, thereby safeguarding cardiomyocytes against damage in DCM. This mechanism establishes a novel theoretical foundation for the clinical treatment of cardiac conditions utilizing MSCs.

Intervertebral disc degeneration (IDD) is a common degenerative disease of the spine that has a significant impact on both society and human health. Many studies have confirmed that there is a close relationship between IDD and senescence and apoptosis, and autophagy can combat apoptosis and senescence. Spatholobi caulis (SC) is an herb that contains various active compounds that are effective in tissue repair and regeneration, but it has not been explored in field of IDD. In this study, it was first found that SC can boost autophagy and reduce the apoptosis and senescence of Nucleus pulposus cell (NPCs). However, our animal studies revealed limited absorption of SC. To improve the bioavailability and efficacy of SC, we developed a hydrogel incorporating quaternary ammonium chitosan (QCS) and oxidized starch (OST) as carriers for SC. The QCS-OST/SC hydrogel exhibits excellent compatibility with cells, can be easily injected, and can release SC durably. At the cellular level, the QCS-OST/SC hydrogel enhances cell viability, initiates autophagy and release of the extracellular matrix (ECM), and inhibits cellular senescence and apoptosis. The injection of the QCS-OST/SC hydrogel via microneedles (MNs) into discs had successfully diminished disc degeneration in rats, which shows that this hydrogel has broad potential in the treatment of IDD.

PANoptosis, a programmed cell death, shares key characteristics of apoptosis, pyroptosis, and necroptosis. Accumulating evidence suggests that PANoptosis plays a crucial role in tumorigenesis. However, the respective regulation mechanisms in cancer are so far unclear. Using various bioinformatic approaches, we comprehensively analyzed the expression patterns, genetic alterations, prognostic value, and immunological role of PANoptosis genes in pan-cancer. Expression of the PANoptosis gene, PYCARD, was validated based on the Human Protein Atlas database and real-time quantitative reverse transcription polymerase chain reaction (RT-PCR). We found that PANoptosis genes were aberrantly expressed in most cancer types, which was consistent with the validation of PYCARD expression. Concurrently, PANoptosis genes and PANoptosis scores were significantly associated with patient survival in 21 and 14 cancer types, respectively. Pathway analysis showed that PANoptosis score was positively correlated with pathways linked to immune and inflammatory responses in pan-cancer, such as IL6-JAK-STAT3 signaling, the interferon-gamma response, and IL2-STAT5 signaling. In addition, the PANoptosis score was significantly correlated with the tumor microenvironment, the infiltration levels of most immune cells (i.e.NK cells, CD8+ T cells, CD4+ T cells, DC cells), and immune-related genes. Furthermore, it was a predictive indicator of immunotherapy response in patients with tumors. These insights substantially improve our understanding of PANoptosis components in cancers and may inspire the discovery of novel prognostic and immunotherapy response biomarkers.

Background:Arthroscopic suture repair is the main treatment option for hip labral tears; however, anchor insertion and placement from arthroscopic portals is difficult. Purpose:To quantitatively evaluate the safety of various arthroscopic portals for suture anchor placement during hip labral repair. Study Design:Descriptive laboratory study. Methods:The computed tomography scans of 20 patients with normally developed hip joints were used to create 3-dimensional models. The distances from the anchor to the articular cartilage (DAC) and from the acetabular insertion point to the cortical bone (DCB) were measured in the anterolateral portal (AL), posterolateral portal (PL), midanterior portal (MAP), medial MAP, and 3 distal anterolateral accessory portals (DALAs): DALA-proximal, DALA-middle, and DALA-distal. Labral tears were divided into anterior (4, 3, and 2 o'clock), lateral (1, 12, and 11 o'clock), and posterior (10, 9, and 8 o'clock) acetabular zones, and the Kruskal-Wallis and Mann-Whitney U test were used to compare DAC and DCB in the zones. The success rate was defined as anchors placed with DAC ≥1 mm and DCB ≥15 mm. Results:The DAC was significantly smaller in the AL at 1 o'clock (0.68 ± 0.32 mm; P < .001) and 12 o'clock (0.37 ± 0.30 mm; P < .001), and in the PL at 12 o'clock (-0.35 ± 0.38 mm; P < .001) and 11 o'clock (0.60 ± 0.24 mm; P < .001). The DCB was significantly smaller in the DALA-P at 3 o'clock (8.93 ± 2.12 mm; P < .001) and 11 o'clock (9.59 ± 2.84 mm; P < .001), the MAP at 12 o'clock (13.76 ± 3.89 mm; P < .001) and 11 o'clock (0.27 ± 0.27 mm; P < .001), and the MMA at 12 o'clock (5.96 ± 2.31 mm; P < .001) and 11 o'clock (0 mm; P < .001). Success rates were high for MAP and MMA between 4 o'clock and 1 o'clock, for DALA-proximal at 12 o'clock, for AL at 11 o'clock, and for PL between 10 o'clock and 8-o'clock. Conclusion:There were significant differences in the success rate of anchor placement using different portals during hip arthroscopic labral repair. Clinical Relevance:MAP is recommended for labral repair between 4 o'clock and 1 o'clock, DALA-P is recommended between 2 o'clock and 12 o'clock, AL is suitable at 11 o'clock, and PL is suitable between 10 o'clock and 8 o'clock.

ABSTRACT Epithelial ovarian cancer (EOC) is the most lethal gynecological cancer, which remains a threat to female health at all ages. Hypotheses for EOC development include the continuous presence of inflammation, in which microbiota and inflammatory cytokines participate in cancer-related signaling pathway activation. Hedgehog (Hh) signaling is prominent for EOC progression, and interacts with inflammation response related to gut microbiota (GM). However, the precise roles of GM during this process are unknown. Here, we showed that the GM from patients with EOC differed from that of healthy women and had GM dysbiosis. We found that EOC modeling may lead to GM changes in mice, and it restored after the administration of GM from healthy controls, while GM from patients with EOC further exacerbated GM dysbiosis. Furthermore, we found that GM from EOC markedly promoted tumor progression and activated Hh signaling; meanwhile, it increased the extent of inflammation and activated NF-κB signaling, but GM from healthy controls improved them. Our results demonstrate how GM dysbiosis promoted EOC progression by activating Hh signaling mediated by TLR4/NF-κB signaling. We anticipate our assay to be a new thought for exploring the role of GM in EOC development. Furthermore, improving GM dysbiosis is a novel therapeutic approach for delaying EOC development.

Epithelial ovarian cancer (EOC), the most predominant subtype of ovarian cancer (OC), involves poor prognosis and exhibits high aggression. Triptolide (TPL), like other Chinese herbs, has historically played a significant role in modern medicine. The screening system based on Gli-dependent luciferase reporter activity assessed the effects of over 800 natural medicinal materials on hedgehog (Hh) signaling pathway activity and discovered that TPL had an excellent inhibitory effect on Hh signaling pathway activity. However, the significance and mechanism of TPL involvement in regulating the Hh pathway have not been well explored. Thus, this work aimed to understand better how TPL affects the Hh pathway activity, which, in turn, influences the biological behavior of EOC. Our findings observed that Smo agonist SAG-induced EOC cell proliferation, migration, and invasion were drastically reversed by TPL in a concentration-dependent pattern. Further evidence suggested that TPL promotes the degradation of Gli1 and Gli2 to inhibit the activity of the Hh signaling pathway by relying on Gli1 and Gli2 ubiquitination. Our in vivo studies also confirmed that TPL could significantly inhibit the tumor growth of EOC. Taken together, our results revealed that one of the antitumor mechanisms of TPL was the targeted inhibition of the Hh/Gli pathway.

Spinal cord injury (SCI) is a serious disorder of the central nervous system with a high disability rate. Long noncoding RNAs (lncRNAs) are reported to mediate many biological processes. The aim of this study was to explore lncRNA and mRNA expression profiles and functional networks after SCI. Differentially expressed genes between SCI model rats and sham controls were identified by microarray assays and analyzed by functional enrichment. Key lncRNAs were identified using a support vector machine- (SVM-) recursive feature elimination (RFE) algorithm. A trans and cis regulation model was used to analyze the regulatory relationships between lncRNAs and their targets. An lncRNA-related ceRNA network was established. We identified 5465 differentially expressed lncRNAs (DE lncRNAs) and 8366 differentially expressed mRNAs (DE mRNAs) in the SCI group compared with the sham group (fold change > 2.0, p < 0.05). Four genes were confirmed by qRT-PCR which were consistent with the microarray data. GSEA analysis showed that most marked changes occurred in pathways related to immune inflammation and nerve cell function, including cytokine-cytokine receptor interaction, neuroactive ligand-receptor interaction, and GABAergic synapse. Enrichment analysis identified 30 signaling pathways, including those associated with immune inflammation response. A total of 40 key lncRNAs were identified using the SVM-RFE algorithm. A key lncRNA-mRNAs coexpression network was generated for 230 951 lncRNA-mRNA pairs with half showing positive correlations. Several key DE lncRNAs were predicted to have "cis"- or "trans"-regulated target genes. The transcription factors, Sp1, JUN, and SOX10, may regulate the interaction between XR_001837123.1 and ETS 1. In addition, five pairs of ceRNA regulatory sequences were constructed. Many mRNAs and lncRNAs were found to be dysregulated after SCI. Bioinformatic analysis showed that DE lncRNAs may play crucial roles in SCI. It is anticipated that these findings will provide new insights into the underlying mechanisms and potential therapeutic targets for SCI.