PURPOSE:Phosphaturic mesenchymal tumor (PMT) is an ultrarare neoplasm responsible for tumor-induced osteomalacia (TIO). Spinal involvement is exceptionally uncommon, and the diagnostic and therapeutic strategies for spinal PMT remain inadequately defined. This report aims to present a case of thoracic vertebral PMT and review the clinical features, diagnostic challenges, and surgical management of spinal PMTs. METHODS:We reported a 60-year-old male patient presenting with recurrent low back pain and persistent hypophosphatemia. Comprehensive imaging, including CT, MRI, and 18F-AlF-NOTA-octreotide PET/CT, was performed. Given the hypervascularity of the lesion, preoperative segmental artery embolization was conducted to reduce the risk of massive intraoperative hemorrhage. En-bloc tumor resection combined with posterior spinal stabilization was subsequently performed. The literature of previously reported spinal PMT cases was also reviewed. RESULTS:Imaging revealed an osteolytic lesion involving the T11 vertebral body and right pedicle with significant contrast enhancement on CT and MRI. 18F-AlF-NOTA-octreotide PET/CT demonstrated intense uptake in the tumor lesion, and histopathology confirmed the diagnosis of classical type PMT. Preoperative embolization effectively reduced intraoperative blood loss, enabling complete tumor resection. Postoperatively, serum phosphorus level normalized within one week, and his symptoms resolved completely. No recurrence was observed during six months of follow-up. CONCLUSION:Spinal PMT, though rare, should be considered in patients with unexplained hypophosphatemia and osteopenia. 18F-AlF-NOTA-octreotide PET/CT is a valuable tool for tumor localization. Preoperative embolization is an effective adjunctive strategy to reduce intraoperative hemorrhage for hypervascular spinal PMTs. Complete surgical resection remains the definitive treatment, offering excellent biochemical and clinical outcomes.

Neoadjuvant chemotherapy has improved survival rates in osteosarcoma patients. However, reliable pretreatment prognostic markers for predicting treatment outcomes remain limited and underexplored. In this multicenter retrospective study, 308 osteosarcoma patients from ZJU hospital (training cohort) and 141 patients from RJ hospital (testing cohort) who underwent neoadjuvant chemotherapy followed by tumor resection were analyzed. Tumor necrosis rates, complete blood count (CBC) parameters, and various clinical features were evaluated. The training cohort was used to develop a predictive model, which was subsequently validated in the testing cohort. Prognostic factors were evaluated using Chi-square, Mann–Whitney U, Cox regression, and log-rank tests. Most tumors (95.5

Endometrial cancer (EC) is a significant health threat to women, with recurrence after treatment posing a major challenge. While abnormal cholesterol metabolism has been implicated in EC progression, the underlying mechanisms remain unclear. In this study, we identified lanosterol synthase (LSS) as a key mediator in cholesterol metabolism associated with EC. We found that LSS is significantly upregulated in EC tissues. Functional assays revealed that LSS promotes cell proliferation and migration, inhibits apoptosis, and drives tumor growth in vivo. Mechanistically, LSS exerts dual effects by accumulating cholesterol esters, thereby enhancing EC cell growth, and activating the MAPK/JNK signaling pathway. Importantly, inhibition of LSS with the specific inhibitor Ro 48-8071 not only reduced EC cell proliferation and suppressed xenograft tumor growth but also inhibited the growth of patient-derived tumor-like cell clusters (PTCs). These findings establish LSS as a novel oncogene in EC, promoting tumor progression through MAPK/JNK signaling activation and cholesterol ester accumulation, and highlight the therapeutic potential of targeting LSS in EC treatment.

BACKGROUND:Mental disorders frequently co-occur with pain, yet pain mechanisms in non-peripheral etiologies (e.g., chronic psychological stress) remain underexplored. The ventral anterior-lateral thalamic complex (VAL) is implicated in emotional processing, but its role in chronic stress-induced pain hypersensitivity is unclear. Electroacupuncture (EA) is clinically used for pain management, but its efficacy and mechanisms in chronic stress-driven pain hypersensitivity require validation. METHODS:A chronic restraint stress (CRS) model was established in male mice. Behavioral assessments were performed to quantify mechanical sensitivity (hindpaws and abdomen using von Frey filaments), thermal sensitivity (hot plate test), and spontaneous pain-like behaviors. Bidirectional chemogenetic approaches targeted VAL CaMKIIα-positive neurons. EA was applied at Zusanli (ST36) and Sanyinjiao (SP6) acupoints. RESULTS:CRS stably induced pain hypersensitivity phenotypes, including mechanical allodynia (hindpaws/abdomen), thermal hyperalgesia, and spontaneous pain-like behaviors. Chemogenetic inhibition of VAL CaMKIIα-positive neurons reversed these CRS-induced hypersensitivity responses. Conversely, activating these neurons in naive mice recapitulated the full spectrum of hyperalgesia phenotypes. EA alleviated CRS-induced hindpaw mechanical/thermal hyperalgesia, abdominal allodynia, and spontaneous pain. EA's effects on hindpaw mechanical/thermal hyperalgesia were mediated by suppression of VAL CaMKIIα-positive neurons. In contrast, its amelioration of abdominal allodynia and spontaneous pain persisted despite chemogenetic activation of VAL CaMKIIα-positive neurons, indicating possible distinct pathways. CONCLUSION:This study reveals the pivotal role of thalamic VAL CaMKIIα-positive neurons in chronic stress-associated pain hypersensitivity and elucidates EA's analgesic mechanisms, providing novel therapeutic strategies for emotion-pain comorbidity.

Osteosarcoma (OS) is considered a sex steroid hormone-dependent bone tumor. The development and progression of OS are regulated by 17β-estradiol (E2). However, the detailed mechanisms of E2-modulated OS progression remained to be elucidated. Here, we found that E2-activated mammalian target of rapamycin (mTOR) signaling promoted N6-methyladenosine (m6A) modification through regulating WTAP. Inhibition of mTOR complex 1 (mTORC1) reversed E2-activated WTAP expression. Meanwhile, inhibition of mTORC1 suppressed OS cell proliferation and migration. Deficiency of TSC2 activated mTORC1 signaling and enhanced OS cell proliferation and migration, while abrogated by Rapamycin. Interestingly, mTOMC1 promoted mRNA stability of ubiquitin-specific protease 7 (USP7) through m6A modification. Loss of USP7 suppressed the proliferation, migration, and ASC specks, while promoted apoptosis of OS cells. USP7 interacted with NLRP3 and deubiquitinated NLRP3 through K48-ubiquitination. USP7 was upregulated and positive correlation with NLRP3 in OS patients with high level of E2. Loss of USP7 suppressed the progression of OS via inhibiting NLRP3 inflammasome signaling pathway. Our results demonstrated that E2-activtated mTORC1 promoted USP7 stability, which promoted OS cell proliferation and migration via upregulating NLRP3 expression and enhancing NLRP3 inflammasome signaling pathway. These results discover a novel mechanism of E2 regulating OS progression and provide a promising therapeutic target for OS progression.

Background: Diabetic kidney disease (DKD), a common complication of diabetes, increasingly threatens patients. Approximately 20%-40% of diabetic patients develop DKD, with 30%-50% progressing to end-stage renal disease. QingReYiQiHuoXue Formula (QRF) has demonstrated clinical efficacy in treating DKD, yet its underlying mechanisms remain unclear. Purpose: This study aimed to explore the mechanisms of QRF in treating DKD and to validate its effects on lipid abnormalities and ferroptosis in a DKD mouse model. Methods: An integrated approach, encompassing network pharmacology, molecular biology, pharmacodynamics, and lipidomics, was employed to elucidate the mechanisms of QRF in treating DKD. Network pharmacology was initially used to predict potential pathways and targets of QRF in treating DKD, along with potential targets for ferroptosis from the ferroptosis database. The efficacy was then validated in a DKD mouse model induced by HFD and STZ injection, including blood and urine tests, histopathological analysis, ferroptosis level measurement, and lipidomic analysis. Results: Through network pharmacology, we discovered that QRF may regulate both the PI3K/AKT/GSK3β signaling pathway and ferroptosis. In vivo experiments confirmed QRF's renal protective effects in HFD/STZ-induced DKD mice, significantly reducing blood glucose levels, ameliorating blood lipid abnormalities, and reducing renal tissue damage. Lipidomic analysis revealed QRF's regulatory effects on renal lipid metabolism, particularly in reducing levels of polyunsaturated fatty acids (PUFAs) associated with ferroptosis. Further validation showed QRF's potential in modulating ferroptosis and the PI3K/AKT/GSK3β signaling pathway. Conclusion: QRF alleviates renal damage in a diabetic mouse model by modulating the PI3K/AKT/GSK3β signaling pathway, improving lipid metabolism disorders, and suppressing ferroptosis. This study offers preliminary insights into the mechanisms of QRF in treating DKD. Disclosure Y. Wei: None. L. Zhang: None.

Digestive tract tumors are heterogeneous and involve the dysregulation of multiple signaling pathways. The Janus kinase-signal transducer and activator of transcription (JAK–STAT) pathway plays a notable role in the oncogenesis of digestive tract tumors. Typically activated by pro-inflammatory cytokines, it regulates important biological processes, such as cell growth, differentiation, apoptosis, immune responses, and inflammation. The aberrant activation of this pathway manifests in different forms, including mutations in JAKs, overexpression of cytokine receptors, and sustained STAT activation, and contributes to promoting the malignant characteristics of cancer cells, including uncontrolled proliferation, resistance to apoptosis, enhanced invasion and metastasis, angiogenesis, acquisition of stem-like properties, and drug resistance. Numerous studies have shown that aberrant activation of the JAK-STAT pathway is closely related to the development and progression of digestive tract tumors, contributing to tumor survival, angiogenesis, changes in the tumor microenvironment, and even immune escape processes. In addition, this signaling pathway also affects the sensitivity of digestive tract tumors to chemotherapy and targeted therapy. Therefore, it is crucial to comprehensively understand the oncogenic mechanisms underlying the JAK-STAT pathway in order to develop effective therapeutic strategies against digestive tract tumors. Currently, several JAK–STAT inhibitors are undergoing clinical and preclinical trials as potential treatments for various human diseases. However, further investigation is required to determine the role of this pathway, as well as the effectiveness and safety of its inhibitors, especially in the context of digestive tract tumors. In this review, we provide an overview of the structure, classic activation, and negative regulation of the JAK-STAT pathway. Furthermore, we discuss the pathogenic mechanisms of JAK-STAT signaling in different digestive tract tumors, with the aim of identifying potential novel therapeutic targets.

Pain-related aversive memory is common in chronic pain patients. Electroacupuncture has been demonstrated to block pain-related aversive memory. The insular cortex is a key region closely related to aversive behaviors. In our study, a potential mechanism underlying the effect of electroacupuncture treatment on pain-related aversive memory behaviors relative to the insular cortex was investigated. Our study used the chemogenetic method, pharmacological method, electroacupuncture intervention, and behavioral detection. Our study showed that both inhibition of gamma-aminobutyric acidergic neurons and activation of the kappa opioid receptor in the insular cortex blocked the pain-related aversive memory behaviors induced by 2 crossover injections of carrageenan in mice; conversely, both the activation of gamma-aminobutyric acidergic neurons and inhibition of kappa opioid receptor in the insular cortex play similar roles in inducing pain-related aversive memory behaviors following 2 crossover injections of carrageenan. In addition, activation of gamma-aminobutyric acidergic neurons in the insular cortex reversed the effect of kappa opioid receptor activation in the insular cortex. Moreover, electroacupuncture effectively blocked pain-related aversive memory behaviors in model mice, which was reversed by both activation of gamma-aminobutyric acidergic neurons and inhibition of kappa opioid receptor in the insular cortex. The effect of electroacupuncture on blocking pain-related aversive memory behaviors may be related to the activation of the kappa opioid receptor and inhibition of gamma-aminobutyric acidergic neurons in the insular cortex.

There is growing evidence that programmed death ligand-1 (PD-L1) has exciting therapeutic efficacy in hematological malignancy and partial solid tumors. However, many patients still face failure with the treatment of immune checkpoint blockade because of PD-L1 expression regulation during transcription and post-transcription processes, including N6-methyladenosine (m6A). Similar to the epigenetic regulation in DNA and histones, recent research has revealed the essential regulation of m6A modification in RNA nuclear export, metabolism and translation. Recent studies have shown that m6A-induced PD-L1 expression emerges as one of the main reasons for the immunological alteration in this process and contributes to the failure of T cell-induced anti-tumor immunity. The results of preclinical studies demonstrate the potential of m6A-targeted therapy in combination with immune checkpoint blockade. The comprehensive expression of m6A-related genes also provided the possibility to indicate the prognosis and to optimize the treatment for patients of various cancer types. In this review, we focus on the m6A modification in PD-L1 mRNA as well as the regulation of PD-L1 expression in cancer cells and summarize its clinical value in anti-PD-L1 cancer immune therapy.

BackgroundThe prognosis of colorectal cancer with atypical metastasis is poor. However, atypical metastasis was less common and under-appreciated. MethodsIn this study we attempted to present the first machine learning models to predict the risk of atypical metastasis in colorectal cancer patients. We evaluated the differences between metastasis and non-metastasis groups, assessed factors associated with atypical metastasis using univariate and multivariate logistic regression analyses, and preliminarily developed the multiple machine learning models to predict atypical metastasis. Results168 patients were included. Prognostic Nutritional Index (PNI) [OR = 0.998; P = 0.030], Cancer antigen 19-9 (CA19-9) [OR = 1.011; P = 0.043] and MR-Distance [-mid OR = 0.289; P = 0.009] [-high OR = 0.248; P = 0.021] were shown to be independent risk factors for the atypical metastasis via multivariate analysis. Furthermore, the machine learning model based on AdaBoost algorithm (AUC: 0736) has better predictive performance comparing to Logistic Regression (AUC: 0.671) and KNeighbors Classifier (AUC: 0.618) by area under the curve (AUC) in the validation cohorts. The accuracy, sensitivity, and specificity of the model trained using the Adaboost method in the validation set are 0.786, 0.776 and 0.700, while 0.601, 0.933, 0.508 using Logistic Regression and 0.743, 0.390, 0.831 using KNeighbors Classifier. ConclusionMachine-learning approaches containing PNI, CA19-9 and MR-Distance show great potentials in atypical metastasis prediction.

ABSTRACT The poor progress of immunotherapy on osteosarcoma patients requires deeper delineation of immune tolerance mechanisms in the osteosarcoma microenvironment and a new therapeutic strategy. Clearance of apoptotic cells by phagocytes, a process termed “efferocytosis,” is ubiquitous in tumors and mediates the suppression of innate immune inflammatory response. Considering the massive infiltrated macrophages in osteosarcoma, efferocytosis probably serves as a potential target, but is rarely studied in osteosarcoma. Here, we verified M2 polarization and PD-L1 expression of macrophages following efferocytosis. Pharmacological inhibition and genetic knockdown were used to explore the underlying pathway. Moreover, tumor progression and immune landscape were evaluated following inhibition of efferocytosis in osteosarcoma model. Our study indicated that efferocytosis promoted PD-L1 expression and M2 polarization of macrophages. Ëfferocytosis was mediated by MerTK receptor in osteosarcoma and regulated the phenotypes of macrophages through the p38/STAT3 pathway. By establishing the murine osteosarcoma model, we emphasized that inhibition of MerTK suppressed tumor growth and enhanced the T cell cytotoxic function by increasing the infiltration of CD8+ T cells and decreasing their exhaustion. Our findings demonstrate that MerTK-mediated efferocytosis promotes osteosarcoma progression by enhancing M2 polarization of macrophages and PD-L1-induced immune tolerance, which were regulated through the p38/STAT3 pathway.

As the main immune checkpoint, PD-L1-PD-1 interaction plays a critical role in the dysregulation of effector T cells, which contributes to the failure of Chimeric Antigen Receptor T-cell (CAR-T) and other immunotherapies. Presently, most research focuses on the extracellular function of PD-L1. Membrane PD-L1 can interact with its receptor PD-1 and decrease T cell-induced cancer immunity. However, the function of PD-L1 in cancer cells is still unclear. Recent studies have shown the separated clinical significance of PD-L1 expression in various cancer types, showing the complexity of PD-L1 in cancer cell regulation. As a novel regulatory pathway, the nuclear translocation of PD-L1 in cancer cells receives more attention. Results of these preclinical studies demonstrated that nuclear PD-L1 has an essential role in cancer development and other immune checkpoint molecules transcription. Herein, we summarized the mechanisms involved in PD-L1 nuclear transportation and identify the key regulatory factors in this process. Furthermore, we also summarize the function of nuclear PD-L1 in cancer immunity. These findings suggested the novel PD-L1 regulation in cancer development, which showed that nuclear PD-L1 is a potential therapeutic target in future cancer therapy.

Research shows that across life, the incidence of mental illness is highest in the young. In the context of the COVID-19 pandemic, mental health issues of the young in particular have received global attention. The rostral anterior cingulate cortex (rACC) plays an important role in psychiatric disorders and chronic pain-psychiatric comorbidities. However, it remains unknown whether or how the afferent and efferent circuits of the rACC change with aging. In this study, we microinjected a retrograde tracer virus and an anterograde trans-monosynaptic virus into the rACC of young and middle-aged mice (both male and female), and systematically and quantitatively analyzed the whole-brain afferent and efferent connections of rACC at different ages and sexes. Notably, in young and middle-aged mice, afferents of the rACC belong to four groups of brain structures arising mainly from the amygdala [mainly basolateral amygdaloid nucleus (BLA)] and cerebral cortex (mainly orbital cortex), with a small part originating from the basal forebrain and thalamus. In contrast, efferents of the rACC belong to four groups of brain structures mainly projecting to the thalamus (mainly ventral anterior-lateral/ventromedial thalamic nucleus (VAL/VM)], with a very small part projecting to the amygdala, basal forebrain, and cerebral cortex. Compared with young mice, the BLA-rACC circuit in middle-aged mice (male and female) did not change significantly, while the rACC-VAL/VM circuit in middle-aged mice (male and female) decreased significantly. In conclusion, this study comprehensively analyzed the input-output neural projections of rACC in mice of different ages and sexes and provided preliminary evidence for further targeted research.

Background: Ferroptosis was reported to have tremendous promise in the treatment and prognosis of hepatocellular carcinoma (HCC). Here, we identified a novel ferroptosis-related prognostic signature incorporating epigenetic and transcriptional biomarkers could help predicting survival of patients with HCC. Methods: We employed multi-omics and clinical data from The Cancer Genome Atlas (TCGA) database to identify the ferroptosis-associated methylation CpG sites associated with HCC survival using sure independence screening (SIS). Then we utilized Kaplan-Meier curves to evaluate the prognostic significance of gene expression and DNA methylation. Receiver operating characteristic (ROC) curve was used predicting the 3- and 5- year survival. Mediation analysis of ferroptosis-related methylation and transcriptional score was performed. Results: We firstly identified 114 significant CpG sites under the criteria of false discovery rate (FDR) <0.05 in training set. Then we screened out 5 candidate CpG sites in validation set for multivariate screening and stepwise regression. We found that the high-risk group had significantly shorter survival time than the low-risk group in the prognostic signature combined with epigenetic and transcriptional scores (HR =2.72 95% CI: 2.01-3.68, P=8.75E-11). And the predictive model involving clinical information, gene expression, and methylation data performed best for 3-year survival prediction (AUC = 0.672) and 5-year survival prediction (AUC =0.742). Conclusions: Our results suggested a signature combining clinical information, ferroptosis-related gene expression, and methylation presented a superior ability for prognostic prediction in HCC, which may bring us novel tool and targets in the treatment of HCC.

Redundancy of multinucleated mature osteoclasts, which results from the excessive fusion of mononucleated preosteoclasts (pOCs), leads to osteolytic diseases such as osteoporosis. Unfortunately, the currently available clinical drugs completely inhibit osteoclasts, thus interfering with normal physiological bone turnover. pOC-specific regulation may be more suitable for maintaining bone homeostasis. Here, circBBS9, a previously unidentified circular RNA, was found to exert regulatory effects via the circBBS9/miR-423-3p/Traf6 axis in pOCs. To overcome the long-standing challenge of spatiotemporal RNA delivery to cells, we constructed biomimetic nanoparticles to achieve the pOC-specific targeted delivery of circBBS9. pOC membranes (POCMs) were extracted to camouflage cationic polymer for RNA interference with circBBS9 (POCM-NPs@siRNA/shRNAcircBBS9). POCM-NPs endowed the nanocarriers with improved stability, accurate pOC targeting, fusogenic uptake, and reactive oxygen species-responsive release. In summary, our findings may provide an alternative strategy for multinucleated cell-related diseases that involves restriction of mononucleated cell multinucleation through a spatiotemporally selective delivery system.

MicroRNAs (miRs) have been implicated in the development and progression of osteosarcoma (OS). However, the underlying mechanism of miR-101 in regulating of the proliferation, migration and invasion of OS cells remains to be elucidated. In the present study, reverse transcription-quantitative polymerase chain reaction data revealed that miR-101 was frequently downregulated in the tissue samples of 12 patients with OS compared with their matched adjacent non-tumor tissues. Furthermore, miR-101 was significantly downregulated in three common OS cell lines, Saos-2, MG63 and U2OS, compared with the human osteoblast cell line, hFOB1.19 (P<0.01). A luciferase reporter assay was also performed and identified c-FOS as a novel target of miR-101 in U2OS cells; overexpression of miR-101 significantly suppressed the protein expression levels of c-FOS, while knockdown of miR-101 significantly enhanced the formers' expression levels in U2OS cells (P<0.01). Independent inhibition of c-FOS and overexpression of miR-101 expression levels significantly suppressed U2OS cell proliferation, migration and invasion (P<0.01). However, overexpression of c-FOS reversed the inhibitory effect of miR-101 upregulation on proliferation, migration and invasion of U2OS cells, suggesting that miR-101 acts as a tumor suppressor in OS cells via targeting of c-FOS. Thus, we propose that the miR-101/c-FOS axis may be a potential therapeutic target for OS.

This study aims to investigate the differentiation trajectory of osteosarcoma cells and to construct molecular subtypes with their respective characteristics and generate a multi-gene signature for predicting prognosis. Integrated single-cell RNA-sequencing (scRNA-seq) data, bulk RNA-seq data and microarray data from osteosarcoma samples were used for analysis. Via scRNA-seq data, time-related as well as differentiation-related genes were recognized as osteosarcoma tumor stem cell-related genes (OSCGs). In Gene Expression Omnibus (GEO) cohort, osteosarcoma patients were classified into two subtypes based on prognostic OSCGs and it was found that molecular typing successfully predicted overall survival, tumor microenvironment and immune infiltration status. Further, available drugs for influencing osteosarcoma via prognostic OSCGs were revealed. A 3-OSCG-based prognostic risk score signature was generated and by combining other clinic-pathological independent prognostic factor, stage at diagnosis, a nomogram was established to predict individual survival probability. In external independent TARGET cohort, the molecular types, the 3-gene signature as well as nomogram were validated. In conclusion, osteosarcoma cell differentiation occupies a crucial position in many facets, such as tumor prognosis and microenvironment, suggesting promising therapeutic targets for this disease.

Tumor microenvironment (TME) formation is a major cause of immunosuppression. The TME consists of a considerable number of macrophages and stromal cells that have been identified in multiple tumor types. CCL2 is the strongest chemoattractant involved in macrophage recruitment and a powerful initiator of inflammation. Evidence indicates that CCL2 can attract other host cells in the TME and direct their differentiation in cooperation with other cytokines. Overall, CCL2 has an unfavorable effect on prognosis in tumor patients because of the accumulation of immunosuppressive cell subtypes. However, there is also evidence demonstrating that CCL2 enhances the anti-tumor capability of specific cell types such as inflammatory monocytes and neutrophils. The inflammation state of the tumor seems to have a bi-lateral role in tumor progression. Here, we review works focusing on the interactions between cancer cells and host cells, and on the biological role of CCL2 in these processes.