Objective To investigate the prevalence of thyroid nodules in the physical examination population in Mianyang region and analyze its association with metabolic syndrome. Methods A retrospective study was conducted on 9 978 individuals who underwent health examinations at our hospital from January 2024 to May 2025. Thyroid examinations were performed using color Doppler ultrasound to analyze the prevalence of thyroid nodules in this population. Clinical data of all subjects were collected, and logistic regression analysis was employed to assess the association between metabolic syndrome and the risk of thyroid nodule development. Results The prevalence of thyroid nodules in the physical examination population of Mianyang region was 17.98% (1 794/9 978). The logistic regression results showed that after adjusting for gender, age, BMI, occupation, consumption of non-iodized salt, staying up late, daily sleep duration, anxiety, and depression, metabolic syndrome (OR=6.593, 95% CI: 3.961-10.975) was associated with thyroid nodules (P<0.05). Conclusion The prevalence of thyroid nodules among the physical examination population in the Mianyang area is 17.98%, and metabolic syndrome remains associated with the risk of thyroid nodules after effectively controlling for confounding factors.

Objective:To construct curved microgrooves with gradient surface roughness on polycaprolactone (PCL) members by regulating alkali etching time and to investigate the synergistic effect of surface roughness and curved microgrooves on the in vitro osteogenic differentiation of mouse pre-osteoblasts (MC3T3-E1), aiming to determine the optimal PCL surface modification strategy. Methods:Soft lithography and melt-casting techniques were used to fabricate PCL membranes with regularly arranged curved microgrooves (CMP). Alkali etching was performed for 24, 48, and 72 h. Groups: smooth PCL (control), CMP (curved microgrooves only), CMP-24 h, CMP-48 h, CMP-72 h (CMP etched for 24, 48, 72 h, respectively). Surface physicochemical properties were characterized: surface morphology was observed by scanning electron microscopy (SEM), surface roughness was measured by atomic force microscopy (AFM), and surface hydrophilicity was evaluated by contact angle measurement. MC3T3-E1 cells were cultured in vitro. Cell adhesion, proliferation, and osteogenic differentiation were assessed using cell counting (CCK-8), immunofluorescence staining, alkaline phosphatase (ALP) and Alizarin red staining with quantification. The mRNA expression levels of osteogenesis-related genes [ALP, collagen type Ⅰ (COL-1), Runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), osteopontin (OPN)] were detected by real-time fluorescence quantitative PCR (RT-qPCR). Results:Curved microgrooves were successfully fabricated on PCL membranes. Alkali treatment improved surface hydrophilicity and increased roughness. The CMP-72 h group exhibited the best hydrophilicity (contact angle: 33.2°±5.5°), with significant differences compared to all other groups (all P<0.05). The CMP-72 h group showed the highest roughness [(59.966±4.729) nm], which was significantly different from all other groups (all P<0.05). CCK-8 results on day 5 showed that both curved microgrooves and roughness promoted cell proliferation: CMP-24 h (0.292±0.003) and CMP-72 h (0.383±0.004) were significantly higher than those in the smooth group (0.270±0.005) (all P<0.05). Immunofluorescence staining revealed that curved microgrooves induced significant contact guidance of cells; this effect weakened with increasing etching time. ALP and Alizarin red staining indicated the deepest osteogenic staining in the CMP-48 h group. Both ALP activity (0.013 021±0.000 032) and Alizarin red quantification (0.290±0.003) were highest in the CMP-48 h group, significantly different from all other groups (all P<0.05). RUNX-2 expression in CMP-24 h and CMP-48 h groups (1.845±0.087 and 1.837±0.027, respectively) was significantly higher than in other groups (all P<0.05), with no significant difference between these two groups ( P>0.05). CMP-48 h group exhibited the highest mRNA expression of all osteogenic genes tested, specifically ALP (2.194±0.028), COL-1 (1.983±0.024), OCN (7.644±0.156), and OPN (2.648±0.031), all significantly greater than other groups (all P<0.05). Conclusions:Both curved microgrooves and surface roughness modification enhance the in vitro osteogenic differentiation of cells on PCL membranes. Among the tested strategies, alkali etching of curved microgrooves for 48 hours (CMP-48h) provided the optimal enhancement of osteogenic capability for MC3T3-E1 cells and represented a promising surface modification strategy for future PCL membranes.

Image 1.

Objective To investigate the mechanism by which Biejiajian Pill(BJJP)regulates ferroptosis in hepatocellular carcinoma(HCC)cells through the p62/Keap1/NRF2 pathway and to provide an experimental basis for its application in the prevention and treatment of HCC.Methods Huh7 HCC cells were divided into a normal control group,a BJJP drug serum group,an erastin(a ferroptosis inducer)group,a BJJP drug serum+erastin group,and BJJP drug serum+ferrostatin-1(Fer-1)(a ferroptosis inhibitor)group.BJJP drug serum was prepared with animals treated with BJJP and CCK-8 assay was performed to determine the optimal concentration and duration of BJJP intervention.The levels of intracellular iron(Fe),reduced glutathione(GSH),lipid peroxides(MDA),and reactive oxygen species(ROS)were measured.Western blot was performed to determine the expression levels of FTH1,GPX4,xCT,SLC40A1,Keapl,p62,and NRF2.JC-1 staining was performed to measure mitochondrial membrane potential,and cell immunofluorescence was performed to determine the expression of p62 and Keap1.Results According to the CCK-8 assay results,the cell inhibition rate was highest when BJJP was administered at a high dose of 2.2 g/kg(P＜0.001).Furthermore,the inhibition rate of Huh7 cells was highest when Huh7 cells were treated with high-dose BJJP drug serum for 48 h.Therefore,the serum concentration of high-dose BJJP and 48 h were selected as the treatment dose and duration for the subsequent experiment.Compared with the control group,the BJJP drug serum group,the erastin group,and the BJJP drug serum+erastin group showed increased iron content,decreased GSH content,increased MDA levels,increased ROS aggregation,decreased FTH1,GPX4,xCT,SLC40A1,p62,and NRF2 contents,increased Keap1 content,and decreased mitochondrial membrane potential(P＜0.05).Conclusion BJJP regulates ferroptosis in Huh7 HCC cells by inhibiting the p62/Keap1/NRF2 pathway,demonstrating potentials as a therapeutic agent for HCC.

Osteosarcoma (OS), chondrosarcoma (CS), and Ewing sarcoma (ES) represent primary malignant bone tumors and pose significant challenges in oncology research and clinical management. Conventional research methods, such as two-dimensional (2D) cultured tumor cells and animal models, have limitations in recapitulating the complex tumor microenvironment (TME) and often fail to translate into effective clinical treatments. The advancement of three-dimensional (3D) culture technology has revolutionized the field by enabling the development of in vitro constructed bone tumor models that closely mimic the in vivo TME. These models provide powerful tools for investigating tumor biology, assessing therapeutic responses, and advancing personalized medicine. This comprehensive review summarizes the recent advancements in research on 3D tumor models constructed in vitro for OS, CS, and ES. We discuss the various techniques employed in model construction, their applications, and the challenges and future directions in this field. The integration of advanced technologies and the incorporation of additional cell types hold promise for the development of more sophisticated and physiologically relevant models. As research in this field continues to evolve, we anticipate that these models will play an increasingly crucial role in unraveling the complexities of malignant bone tumors and accelerating the development of novel therapeutic strategies.
Bone Neoplasms/pathology* ; Humans ; Osteosarcoma/pathology* ; Tumor Microenvironment ; Sarcoma, Ewing/pathology* ; Chondrosarcoma/pathology* ; Animals ; Cell Culture Techniques/methods* ; Cell Culture Techniques, Three Dimensional/methods* ; Cell Line, Tumor

Immune suppression in the tumor microenvironment (TME) is closely related to abnormal glycolysis. Tumor cells gain metabolic advantages and suppress immune responses through the "Warburg effect". Traditional Chinese medicine (TCM) has been shown to regulate key glycolysis enzymes (such as HK2 and PKM2), metabolic signaling pathways (such as PI3K/AKT/mTOR, HIF-1α) and non-coding RNAs at multiple targets, thus synergistically inhibiting lactate accumulation, improving vascular abnormalities, and relieving metabolic inhibition of immune cells. Studies have shown that TCM monomers and formulas can promote immune cell infiltration and functions, improve metabolic microenvironment, and with the assistance by the nano-delivery system, enhance the precision of treatment. However, the dynamic mechanism of the interaction between TCM-regulated glycolysis and TME has not been fully elucidated, for which single-cell sequencing and other technologies provide important technical support to facilitate in-depth analysis and clinical translational research. Future studies should be focused on the synergistic strategy of "metabolic reprogramming-immune activation" to provide new insights into the mechanisms of tumor immunotherapy.
Humans ; Tumor Microenvironment/immunology* ; Glycolysis/drug effects* ; Neoplasms/drug therapy* ; Medicine, Chinese Traditional ; Signal Transduction ; Drugs, Chinese Herbal/pharmacology*

Recent studies have shown that the physiological homeostasis of oral mucosal cells is regulated by the circadian clock.Dis-ruption or dysfunction of the circadian clock is closely associated with the development of oral squamous cell carcinoma(OSCC).Research based on the circadian clock offers a novel perspective on the pathogenesis and therapeutic strategies for OSCC.However,there is current-ly limited research on this topic,and people generally have insufficient understanding and recognition of the circadian clock.Given the complexity and challenges of circadian clock which is the fourth dimension of medical research,we organize relevant experts based on summarizing the current research results of circadian clock in the pathogenesis and precision diagnosis and treatment of OSCC,combining the scientific principles of the circadian clock's role and their long-term research experience,then summarizes and recommends the con-sensus opinions for the research of circadian clock in the pathogenesis mechanism and precision diagnosis and treatment of human OSCC,with the hope of providing guidance for the basic research and clinical application of circadian clock or circadian rhythm in the pathogene-sis mechanism and precision diagnosis and treatment of oral and maxillofacial squamous cell carcinoma.

Recent studies have shown that the physiological homeostasis of oral mucosal cells is regulated by the circadian clock.Dis-ruption or dysfunction of the circadian clock is closely associated with the development of oral squamous cell carcinoma(OSCC).Research based on the circadian clock offers a novel perspective on the pathogenesis and therapeutic strategies for OSCC.However,there is current-ly limited research on this topic,and people generally have insufficient understanding and recognition of the circadian clock.Given the complexity and challenges of circadian clock which is the fourth dimension of medical research,we organize relevant experts based on summarizing the current research results of circadian clock in the pathogenesis and precision diagnosis and treatment of OSCC,combining the scientific principles of the circadian clock's role and their long-term research experience,then summarizes and recommends the con-sensus opinions for the research of circadian clock in the pathogenesis mechanism and precision diagnosis and treatment of human OSCC,with the hope of providing guidance for the basic research and clinical application of circadian clock or circadian rhythm in the pathogene-sis mechanism and precision diagnosis and treatment of oral and maxillofacial squamous cell carcinoma.

Objective:To construct curved microgrooves with gradient surface roughness on polycaprolactone (PCL) members by regulating alkali etching time and to investigate the synergistic effect of surface roughness and curved microgrooves on the in vitro osteogenic differentiation of mouse pre-osteoblasts (MC3T3-E1), aiming to determine the optimal PCL surface modification strategy. Methods:Soft lithography and melt-casting techniques were used to fabricate PCL membranes with regularly arranged curved microgrooves (CMP). Alkali etching was performed for 24, 48, and 72 h. Groups: smooth PCL (control), CMP (curved microgrooves only), CMP-24 h, CMP-48 h, CMP-72 h (CMP etched for 24, 48, 72 h, respectively). Surface physicochemical properties were characterized: surface morphology was observed by scanning electron microscopy (SEM), surface roughness was measured by atomic force microscopy (AFM), and surface hydrophilicity was evaluated by contact angle measurement. MC3T3-E1 cells were cultured in vitro. Cell adhesion, proliferation, and osteogenic differentiation were assessed using cell counting (CCK-8), immunofluorescence staining, alkaline phosphatase (ALP) and Alizarin red staining with quantification. The mRNA expression levels of osteogenesis-related genes [ALP, collagen type Ⅰ (COL-1), Runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), osteopontin (OPN)] were detected by real-time fluorescence quantitative PCR (RT-qPCR). Results:Curved microgrooves were successfully fabricated on PCL membranes. Alkali treatment improved surface hydrophilicity and increased roughness. The CMP-72 h group exhibited the best hydrophilicity (contact angle: 33.2°±5.5°), with significant differences compared to all other groups (all P<0.05). The CMP-72 h group showed the highest roughness [(59.966±4.729) nm], which was significantly different from all other groups (all P<0.05). CCK-8 results on day 5 showed that both curved microgrooves and roughness promoted cell proliferation: CMP-24 h (0.292±0.003) and CMP-72 h (0.383±0.004) were significantly higher than those in the smooth group (0.270±0.005) (all P<0.05). Immunofluorescence staining revealed that curved microgrooves induced significant contact guidance of cells; this effect weakened with increasing etching time. ALP and Alizarin red staining indicated the deepest osteogenic staining in the CMP-48 h group. Both ALP activity (0.013 021±0.000 032) and Alizarin red quantification (0.290±0.003) were highest in the CMP-48 h group, significantly different from all other groups (all P<0.05). RUNX-2 expression in CMP-24 h and CMP-48 h groups (1.845±0.087 and 1.837±0.027, respectively) was significantly higher than in other groups (all P<0.05), with no significant difference between these two groups ( P>0.05). CMP-48 h group exhibited the highest mRNA expression of all osteogenic genes tested, specifically ALP (2.194±0.028), COL-1 (1.983±0.024), OCN (7.644±0.156), and OPN (2.648±0.031), all significantly greater than other groups (all P<0.05). Conclusions:Both curved microgrooves and surface roughness modification enhance the in vitro osteogenic differentiation of cells on PCL membranes. Among the tested strategies, alkali etching of curved microgrooves for 48 hours (CMP-48h) provided the optimal enhancement of osteogenic capability for MC3T3-E1 cells and represented a promising surface modification strategy for future PCL membranes.

Objective:To investigate the effect of IL-37 on oxidized low-density lipoprotein(ox-LDL)-induced calcification in human aortic vascular smooth muscle cells(HA-SMCs)and the influence of toll-like receptor(TLR)/nuclear transcription factor(NF-κB)pathway and osteogenic transcription factors,to demonstrate the mechanism of IL-37 in calcification.Methods:The calcifica-tion model of HA-SMCs was treated with ox-LDL,and the model group was pretreated with 100 ng/ml IL-37 or 100 μmol/L PDTC.CCK8 was used to detect cell proliferation activity;flow cytometry was used to detect apoptosis.Alizarin red staining was used to detect calcified nodules.The mRNA levels of smooth muscle actin(SMA),bone morphogenetic protein(BMP2)and IL-37 were detected by qPCR;mRNA and protein expression levels of TLR,NF-κB,p-NF-κB,RUNX-associated transcription factor(RUNX2),alkaline phosphatase(ALP),SMA and smooth muscle 22α(SM22α)were detected by qPCR and Western blot.Results:Compared with the control group,ox-LDL treatment enhanced cellular proliferation activity and was concentration-dependent(P<0.05),and apoptosis rate was increased,while the model group produced significant orange-red calcified nodules,up-regulated mRNA and protein expres-sion of inflammatory factors such as TLR and NF-κB,up-regulated mRNA expression of IL-37,RUNX2,BMP2,ALP,while SMA and SM22α mRNA expressions were decreased,and the protein expressions of p-NF-κB and RUNX2 were increased significantly;compared with the modeling group,the IL-37 pretreatment group showed lower viability and decreased apoptosis rate(P<0.05),sig-nificantly reduced calcified nodules,and the mRNA and protein expression of inflammatory factors such as TLR and NF-κB,and mRNA of RUNX2,BMP2,ALP expressions were downregulated,mRNA expression of SMA and SM22α were elevated,and protein expressions of p-NF-κB and RUNX2 were significantly decreased,showing the same trend as the inhibitor PDTC group in cytokines.Conclusion:IL-37 inhibits calcification of HA-SMCs,which may be related to the inhibition of NF-κB pathway and osteogenic pheno-type transformation.