Craniofacial bone regeneration remains a major clinical challenge, yet the identity of orofacial mesenchymal stem/stromal cells (OMSCs) has not been fully elucidated. Here, we performed single-cell RNA sequencing (scRNA-seq) on mouse orofacial bone and identified multiple stromal cell clusters. Cell-cell communication mapping and trajectory inference uncovered the heterogeneity of OMSCs and functional divergence among subpopulations. We identified a previously unrecognized population, Smmhc-expressing mesenchymal stem/stromal cells (MSCs), at the earliest stage of the progenitor lineage trajectory. In vivo lineage tracing demonstrated that Smmhc+ MSCs are multipotent, giving rise to osteoblasts, osteocytes, periodontal ligament (PDL) cells, and dental pulp cells. Targeted ablation of Smmhc+ MSCs using SmmhcCreER;iDTR mouse model led to impaired orofacial bone development and disrupted orofacial tissue homeostasis, characterized by reduced osteogenic differentiation and non-cell autonomous reduction of bone resorption. Collectively, this study establishes a cellular atlas of OMSCs and identifies Smmhc+ MSCs as a functionally indispensable subset for craniofacial bone homeostasis, orchestrating the dynamic balance between osteogenesis and bone resorption within the orofacial skeletal niche.

Patients with relapsed or refractory (R/R) large B-cell lymphoma (LBCL) who experience disease progression following anti-CD19 chimeric antigen receptor T-cell (CAR-T) therapy face poor prognoses and limited therapeutic options. Bispecific antibodies (BsAbs) have emerged as a promising salvage strategy. This meta-analysis was conducted to evaluate the efficacy and safety of CD3×CD20 BsAbs in R/R LBCL patients after CAR-T failure. Clinical studies published between 2021 and 2025 were systematically reviewed, and a random-effects model was applied for pooled and subgroup analyses. A total of fifteen studies involving 1,169 patients were included. The pooled overall response rate (ORR) was 45

Aim or purpose: Glucose metabolism plays a pivotal role in regulating a wide array of cellular physiological activities, encompassing osteogenic differentiation. Despite this, the potential benefits of targeting energy metabolism for enhancing bone regeneration remain largely unexplored. In this study, we delved into the impact and underlying mechanisms of the salt-induced kinase 2/3 (SIK 2/3) inhibitor, YKL-05-099 (YKL), on mandibular bone defects. Materials and methods: We generated mandibular defect models with YKL local application and analyzed the new bone formation. Orofacial bone-marrow-derived mesenchymal stem cells (OMSCs) were cultured to evaluate the function and mechanism of YKL in osteogenic differentiation. Results: YKL successfully enhanced new bone formation within the mandibular defect area without causing any organ damage. Furthermore, it promoted osteogenic differentiation of OMSCs by inhibiting SIK 2/3, which was accompanied by an increase in glucose uptake and ATP production. Mechanistically, glucose transporter 4 (Glut4) acted as a modulator of osteogenic differentiation by limiting AMPK pathway and preventing RUNX2 ubiquitinated degradation, and SIK 2/3 suppresses Glut4’s expression through the HDAC4/PGC-1α pathway. Conclusions: Collectively, our findings demonstrated YKL can promote osteogenic ability of OMSCs through Glut4-dependent glucose uptake, and SIK 2/3 negatively regulated level of Glut4 via HDAC4/PGC-1α pathway. These insights not only uncover a novel mechanism but also provide a solid foundation for the application of YKL in the context of mandibular bone defects.

BACKGROUND:Breast cancer (BRCA) remains a significant global health concern, with the need for novel therapeutic targets to improve patient outcomes. The role of the SENP family of de-SUMOylating enzymes in BRCA is not yet fully understood. METHODS:The expression and prognostic value of SENP family in BRCA were analyzed using the TCGA database. GSEA was conducted to identify correlations between SENP5 expression and cell cycle pathways. Experiments including Western blotting, RT-qPCR, CCK8 assays, colony formation assays, EdU staining, wound healing assays, and transwell assays were used to assess the impact of SENP5 knockdown on BRCA cell proliferation, migration, and invasion. Co-immunoprecipitation and fluorescence co-localization studies were employed to investigate the interaction between SENP5 and CDK1. The effects of combining SENP5 knockdown with CDK1 inhibition were evaluated in MDA-MB-231 xenograft mouse model. RESULTS:SENP5 was found to be overexpressed in BRCA and associated with poor prognosis. Knockdown of SENP5 significantly inhibited BRCA cell proliferation and migration. GSEA revealed a strong correlation between SENP5 and the cell cycle, particularly the G2M checkpoint and E2F target pathways. SENP5 was shown to promote cell cycle progression by upregulating CDK1. Mechanistically, SENP5 mediates the de-SUMOylation of CDK1, reducing its degradation via the ubiquitin-proteasome pathway and increasing CDK1 expression. In vivo, the combination of SENP5 knockdown and CDK1 inhibition significantly suppressed BRCA tumor growth. CONCLUSION:Our research identifies the SENP5/CDK1 axis as a key player in BRCA progression, highlighting its potential as a therapeutic target.

Targeting CD3 × CD20 bispecific antibodies (BsAbs) represents a new milestone in the salvage therapy of relapsed/relapsed large B cell lymphoma and follicular lymphoma. However, cytokine release syndrome (CRS) remains one of the major concerns in clinical practice of CD3 × CD20 BsAbs. This study aimed to identify the potential predictive factors and to construct a nomogram of grade ≥ 2 CRS in CD3 × CD20 BsAbs in Chinese patients. A total of 87 consecutive patients with B-NHL who received CD3 × CD20 BsAbs at Sun Yat-sen University Cancer Center from January 2021 to December 2023 were included and analyzed. Clinical data were collected, and various methods including machine learning algorithms were introduced. The median age of the patients was 55 years, and the median number of previous treatment lines was 2. CRS occurred in 42 patients (48.3

Hepatocellular carcinoma (HCC) remains a malignant and life-threatening tumor with an extremely poor prognosis, posing a significant global health challenge. Despite the continuous emergence of novel therapeutic agents, patients exhibit substantial heterogeneity in their responses to anti-tumor drugs and overall prognosis. The pentose phosphate pathway (PPP) is highly activated in various tumor cells and plays a pivotal role in tumor metabolic reprogramming. This study aimed to construct a model based on PPP-related Genes for risk assessment and prognosis prediction in HCC patients. We integrated RNA-seq and microarray data from TCGA, GEO, and ICGC databases, along with single-cell RNA sequencing (scRNA-seq) data obtained from HCC patients via GEO. Based on the "Seurat" R package, we identified distinct gene clusters related to the PPP within the scRNA-seq data. Using a penalized Cox regression model with least absolute shrinkage and selection operator (LASSO) penalties, we constructed a risk prognosis model. The validity of our risk prognosis model was further confirmed in external cohorts. Additionally, we developed a nomogram capable of accurately predicting overall survival in HCC patients. Furthermore, we explored the predictive potential of our risk model within the immune microenvironment and assessed its relevance to biological function, particularly in the context of immunotherapy. Subsequently, we performed in vitro functional validation of the key genes (ATAD2 and SPP1) in our model. A ten-gene signature associated with the PPP was formulated to enhance the prediction of HCC prognosis and anti-tumor treatment response. Following this, the ROC curve, nomogram, and calibration curve outcomes corroborated the model's robust clinical predictive capability. Functional enrichment analysis unveiled the engagement of the immune system and notable variances in the immune infiltration landscape across the high and low-risk groups. Additionally, tumor mutation frequencies were observed to be elevated in the high-risk group. Based on our analyses, the IC50 values of most identified anticancer agents demonstrated a correlation with the RiskScore. Additionally, the high-risk and low-risk groups exhibited differential sensitivity to various drugs. Cytological experiments revealed that silencing ATAD2 or SPP1 suppresses malignant phenotypes, including viability and migration, in liver cancer cells. In this study, a novel gene signature related to the PPP was developed, demonstrating favorable predictive performance. This signature holds significant guiding value for assessing the prognosis of HCC patients and directing individualized treatment strategies.

Chemotherapy resistance drives bladder cancer (BC) recurrence and metastasis, but the biomarkers and mechanisms of chemotherapy sensitivity are not fully known. We identified differentially expressed genes (DEGs) in chemo-resistant and -sensitive BC patients from TCGA and GEO databases. Analyses like GO, KEGG, random survival forest were conducted. We studied the relationships of hub genes with immune cell infiltration, pathways, drug sensitivity, prognosis, regulation, and cellular heterogeneity using multiple methods. A total of 4042 up-regulated and 1355 down-regulated DEGs were included in the analysis. Four hubs, RNF19A, PCGF5, UNC5CL, and CCDC146, were identified and linked to tumor immune infiltration, immune-related genes, sensitivity to chemotherapeutic drugs, and the expression of disease-related genes like APC and EGFR. GSVA and GSEA analysis revealed varying expression levels of these genes impacting cancer-related signaling pathways. A nomogram and calibration curves based on these hub genes showed excellent prognosis predictive performance. We identified key binding motifs and transcription factors for hub genes using RcisTarget. Our mRNA-miRNA regulatory network and single cell analysis revealed cellular heterogeneity in hub gene expression. Therefore, up-regulation of RNF19A, PCGF5, UNC5CL, and CCDC146 in BC is associated with chemotherapy response and various cellular functions, making them potential predictive biomarkers for chemotherapy sensitivity and prognosis.

Immune checkpoint blockade therapy targeting the programmed death-1(PD-1) pathway has shown remarkable efficacy and durable response in patients with various cancer types. Early prediction of therapeutic efficacy is important for optimizing treatment plans and avoiding potential side effects. In this work, we developed an efficient machine learning prediction method using routine hematologic and biochemical parameters to predict the efficacy of PD-1 combination treatment in Pan-Cancer patients. A total of 431 patients with nasopharyngeal carcinoma, esophageal cancer and lung cancer who underwent PD-1 checkpoint inhibitor combination therapy were included in this study. Patients were divided into two groups: progressive disease (PD) and disease control (DC) groups. Hematologic and biochemical parameters were collected before and at the third week of PD-1 therapy. Six machine learning models were developed and trained to predict the efficacy of PD-1 combination therapy at 8–12 weeks. Analysis of 57 blood biomarkers before and after three weeks of PD-1 combination therapy through statistical analysis, heatmaps, and principal component analysis did not accurately predict treatment outcome. However, with machine learning models, both the AdaBoost classifier and GBDT demonstrated high levels of prediction efficiency, with clinically acceptable AUC values exceeding 0.7. The AdaBoost classifier exhibited the highest performance among the 6 machine learning models, with a sensitivity of 0.85 and a specificity of 0.79. Our study demonstrated the potential of machine learning to predict the efficacy of PD-1 combination therapy based on changes in hematologic and biochemical parameters.

Oral inflammatory diseases such as apical periodontitis are common bacterial infectious diseases that may affect the periapical alveolar bone tissues. A protective process occurs simultaneously with the inflammatory tissue destruction, in which mesenchymal stem cells (MSCs) play a primary role. However, a systematic and precise description of the cellular and molecular composition of the microenvironment of bone affected by inflammation is lacking. In this study, we created a single-cell atlas of cell populations that compose alveolar bone in healthy and inflammatory disease states. We investigated changes in expression frequency and patterns related to apical periodontitis, as well as the interactions between MSCs and immunocytes. Our results highlight an enhanced self-supporting network and osteogenic potential within MSCs during apical periodontitis-associated inflammation. MSCs not only differentiated toward osteoblast lineage cells but also expressed higher levels of osteogenic-related markers, including Sparc and Col1a1. This was confirmed by lineage tracing in transgenic mouse models and human samples from oral inflammatory-related alveolar bone lesions. In summary, the current study provides an in-depth description of the microenvironment of MSCs and immunocytes in both healthy and disease states. We also identified key apical periodontitis-associated MSC subclusters and their biomarkers, which could further our understanding of the protective process and the underlying mechanisms of oral inflammatory-related bone disease. Taken together, these results enhance our understanding of heterogeneity and cellular interactions of alveolar bone cells under pathogenic and inflammatory conditions. We provide these data as a tool for investigators not only to better appreciate the repertoire of progenitors that are stress responsive but importantly to help design new therapeutic targets to restore bone lesions caused by apical periodontitis and other inflammatory-related bone diseases.

Hypoparathyroidism (HypoPT) is a rare disease involving the parathyroid glands that is characterized by a reduced secretion or potency of the parathyroid hormone (PTH), which leads to high serum phosphorus levels and low serum calcium levels. HypoPT most commonly results from accidental damage to the glands or their removal during thyroid or other anterior neck surgery. Parathyroid/thyroid surgery has become more common in recent years, with a corresponding rise in the occurrence of HypoPT as a postoperative complication. There is a critical need for a HypoPT animal model to better understand the mechanisms underlying the effects of HypoPT on mineral ion homeostasis and to verify the therapeutic effectiveness of novel treatments. Here, a technique is reported to create acquired HypoPT in male rats by performing parathyroidectomy (PTX) using carbon nanoparticles. The rat model shows great promise over the mouse models of hypoparathyroidism. Importantly, the human PTH receptor binding region has an 84.2% sequence similarity with that of the rat, which is higher than the 73.7% similarity shared with mice. Moreover, the effects of estrogen, which can affect the PTH/PTHrP receptor signaling pathway, have not been fully investigated in male rats. Carbon nanoparticles are lymphatic tracers that stain the thyroid lymph nodes black without affecting their function, but they do not stain the parathyroid glands, which makes them easy to identify and remove. In this study, serum PTH levels were undetectable after PTX, and this resulted in significant hypocalcemia and hyperphosphatemia. Thus, the clinical state of postoperative HypoPT can be remarkably represented in the rat model. Carbon-nanoparticle-assisted PTX can, therefore, serve as an extraordinarily effective and readily implementable model for studying the pathogenesis, treatment, and prognosis of HypoPT.

Figure S2. PITX2 is a direct target of miR-644a in ESCC. A, miR-644a and its putative binding sequence in the 3'-UTR of PITX2. The mutant miR-644a-binding site was generated in the complementary site for the seed region of miR-644a (wt: wild type; mt: mutant type). B, miR-644a significantly suppressed the luciferase activity that carried wt but not mt 3'-UTR of PITX2. C and D, up-regulation of miR-644a significantly reduced PITX2 protein (C) and mRNA levels (D) of KYSE-140 and Eca-109 cells. E, PITX2 expression in subcutaneous implantation models of KYSE-140 and Eca-109 cells detected by IHC staining. **P<0.01. *P<0.05.

Dysregulation of non-coding RNAs, including miRNAs and lncRNAs has been reported to play vital roles in gastric cancer (GC) carcinogenesis, but the mechanism involved is largely unknown. Using the cancer genome atlas (TCGA) data set and bioinformatics analyses, we identified miR-532-5p as a potential tumor suppressor in GC, and found that lncRNA LINC01410 might be a negative regulator of miR-532-5p. We then conducted a series of in vivo and in vitro assays to explore the effect of LINC01410 on miR-532-5p-mediated GC malignancy and the underlying mechanism involved. MiR-532-5p overexpression inhibited GC metastasis and angiogenesis in vitro and in vivo, whereas miR-532-5p silencing had the opposite effect. Further study showed that miR-532-5p attenuated NF-κB signaling by directly inhibiting NCF2 expression, while miR-532-5p silencing in GC enhanced NF-κB activity. Furthermore, we demonstrated miR-532-5p down-regulation was caused by aberrantly high expression of LINC01410 in GC. Mechanistically, overexpression of LINC01410 promoted GC angiogenesis and metastasis by binding to and suppressing miR-532-5p, which resulted in up-regulation of NCF2 and sustained NF-κB pathway activation. Interestingly, NCF2 could in turn increase the promoter activity and expression of LINC01410 via NF-κB, thus forming a positive feedback loop that drives the malignant behavior of GC. Finally, high expression of LINC01410, along with low expression of miR-532-5p, was associated with poor survival outcome in GC patients. Our studies uncover a mechanism for constitutive LINC1410-miR-532-5p-NCF2-NF-κB feedback loop activation in GC, and consequently, as a potential therapeutic target in GC treatment.

MiR-644a inhibits the stemness of Eca-109 cells by regulating PITX2 expression. A-D, enforced overexpression of miR-644a in Eca-109 cells substantially down-regulated the levels of stemness-associated genes (Nanog, Oct-4, Bmi-1, Notch-1 and Smo), multiple drug-resistance transporter genes (ABCC2, ABCG2) and surface antigens associated with cancer stem cells (CD24, CD44, CD133, CD105, and CD166) (A), reduced phere-forming ability (B) and proportion of side-population cells (C), and also largely increased chemosensitivity to cisplatin or radiosensitivity to IR (D). Restoration of PITX2 in miR-644a-overexpressing Eca-109 cells largely rescued the cells' stemness, while knockdown of PITX2 by shPITX2 decreased Eca-109 cells' stemness (A-D). E, tumor formation in nude mice shows reduced tumorigenicity in 3 groups of miR-644a-overexpressing Eca-109 cells indicated, as compared to that in matched control groups. *P<0.05, **P<0.01.

Background: Dexmedetomidine is a widely used anaesthetic adjuvant for cancer resection surgeries. However, recent reports suggest that it may promote tumour growth or metastasis, so it is essential to clarify its tumour-related effects.Methods: Seven syngeneic murine tumour models were used to assess the impact of dexmedetomidine on primary tumour growth, spontaneous tumour metastasis, and surgical resection-associated metastasis. Cancer cell proliferation and apoptosis experiments, terminal deoxynucleotidyl transferase dUTP nick-end labelling assays, immune cell analysis, specific T-cell depletion experiments, and gene transcription analysis were conducted to identify the underlying mechanisms.Results: Dexmedetomidine did not affect growth of EO771 or 4T1 breast tumours, LAP0297 or LLC lung tumours, MCA205 fibrosarcoma, or their spontaneous lung metastases. It did not promote lung metastasis after breast cancer resection. Dexmedetomidine significantly suppressed MCA38 and CT26 colorectal tumour growth (P<0.01) and promoted apoptosis in MCA38 tumour tissues (P<0.05) without affecting proliferation and apoptosis of MCA38 tumour cells in vitro, suggesting indirect anti-tumour effects. Dexmedetomidine increased the proportions of intratumour CD4+ T (P<0.01), CD8+ T (P<0.001), and natural killer cells (P<0.01), and it upregulated transcription of the cytotoxicity-related genes Infg, Tnfa, and Cxcl9 (P<0.05) in MCA38 tumours. Either CD8+ or CD4+ T-cell depletion reversed the anti-tumour effects of dexmede-tomidine on MCA38 tumours (P<0.05).Conclusions: Dexmedetomidine conferred colorectal tumour-type specific suppression by modulation of tumour CD4+ and CD8+ T cells without tumour-enhancing effects.

Correlation between the expressions of miR-644a and pluripotency-associated factors in 20 fresh ESCC tissues. A, real-time PCR analysis of the mRNA expression of pluripotency-associated factors, including ABCG2, SOX2, OCT4, NANOG, BMI1, and CD44 in 20 ESCC samples. B, the levels of miR-644a were negatively correlated with the expression levels of pluripotency-associated factors in 20 ESCC tissues. **P<0.01, *P<0.05.

Vascular endothelial cells (VECs) injury is the first step in the pathogenesis of atherosclerosis (AS). Mitochondrial dysfunction plays a significant role in VECs injury, but the underlying mechanisms are still unclear. Here, the human umbilical vein endothelial cells were exposed to 100 μg/mL oxidized low-density lipoprotein for 24 h to establish AS model in vitro. We reported that mitochondrial dynamics disorder is a prominent feature of VECs in AS models and associated with mitochondrial dysfunction. Moreover, the knockdown of dynamin-related protein 1 (DRP1) in AS model significantly alleviated the mitochondrial dynamics disorder and VECs injury. On the contrary, DRP1 overexpression significantly aggravated this injury. Interestingly, atorvastatin (ATV), a classical anti-atherosclerotic drug, prominently inhibited the expression of DRP1 in AS models and similarly alleviated the mitochondrial dynamics disorder and VECs injury in vitro and in vivo. At the same time, we found that ATV alleviated VECs damage but did not significantly reduce lipid concentration in vivo. Our findings provide a potential therapeutic target of AS and a new mechanism of the anti-atherosclerotic effect of ATV.

Supplementary Methods, Supplementary Figure legends and Supplementary Table S1 to S4.

The effect of miR-644a on cell growth, migration and invasion of KYSE-410 cell line in vitro. A and B, down-regulation of miR-644a significantly promoted cell growth (A) and colony formation (B) compared with that of control cells. C, the wound healing rate in anti-miR-644a-transfected cells was significantly increased. D, the number of invaded cell in anti-miR-644a-transfected group was significantly increased compared to the control group. **P<0.01, *P<0.05.

Asparaginase/pegaspargase containing regimens combined with radiotherapy are highly effective and considered the cornerstone of localized Natural killer/T-cell lymphoma (NKTL) treatment. However, these chemotherapy regimens inevitably cause relatively high incidence of treatment-related adverse events (TRAEs). Herein we retrospectively evaluated the efficacy and safety of the combined regimen of anti-PD-1 antibody, anlotinib and pegaspargase "sandwich" with radiotherapy in localized NKTL. Anti-PD-1 antibody and pegaspargase at 2500 U/m(2) were administered on day 1, while anlotinib (12 mg once a day) was orally administered on days 1-14. The treatment was repeated every 3 weeks. All the eight patients included received 3 cycles of the regimen followed by radiotherapy and an additional 3 cycles. The overall response rate was 100%, and the complete response rate was 87.5%. With a median follow-up time of 35.5 months (range, 34.03-40.90 months), median PFS and OS times were not reached. The 3-year PFS and OS rates were 100% and 100%, respectively. All patients were alive at the last follow-up. No treatment-related death and no grade 4 TRAE was reported. No grade 3/4 hematological toxicity was detected, and half of the patients didn't report any hematological toxicity. This study indicates that anti-PD-1 antibody combined with anlotinib and pegaspargase is a promising chemoradiotherapy regimen for localized NTKL, with mild toxicity and good tolerance.

Background This study aimed to explore the expression and clinical implication of guanine nucleotide-binding protein alpha 13 (GNA13) in esophageal squamous cell carcinoma (ESCC). Methods We first employed western blot analysis to test the GNA13 protein expression level in ESCC tissues. Subsequently, we used immunohistochemistry assays to detect the GNA13 in ESCC specimens from 173 patients who underwent esophagectomy. Survival analysis was performed to define the impact of GNA13 expressions on the prognosis of the ESCC patients based on the clinical and follow-up data. Results The GNA13 protein was shown to be considerably higher in ESCC tissues than in normal esophageal tissues. The level of expression was closely related to the tumor, node, TNM stage, and tumor size. More importantly, ESCC patients with high GNA13 expression carried an increased risk of tumor recurrence compared to those with low GNA13 expression. In addition, a high GNA13 expression level could independently predict worse overall survival and disease-free survival in ESCC. Conclusions GNA13 could be a novel prognostic biomarker for ESCC patients after esophagectomy.