Objective:To observe the effect of hyperthermia combined with iron overload on the regulation of tumor-associated macrophage polarization and the migration ability of CAL-27 cells and explore its mechanism.Methods:Human monocytic leukemia cell THP-1 was induced and polarized into M2 macrophages. M2 tumor-associated macrophages and CAL-27 cells were divided into the control group (no intervention), hyperthermia group (incubated at 42 ℃ for 1 h, and then incubated at 37 ℃ for 24 h), ferric citrate group (added with 2.5 mg/ml ferric citrate, and cultured in an incubator at 37 ℃for 24 h) and hyperthermia + ferric citrate group (added with 2.5 mg/ml ferric citrate for 1 h, cultured in an incubator at 42℃ for 1 h, and then incubated at 37℃ for 24 h). For M2 macrophage groups, the mRNA relative expression levels of surface markers of M1 macrophage polarization including interleukin (IL)-1β, tumor necrosis factor-α(TNF-α), and those of M2 macrophage polarization including IL-10 and transforming growth factor-β(TGF-β) were detected by real-time reverse transcription polymerase chain reaction. The expression levels of IL-1β and IL-10 were detected by ELISA. The expression levels of CD86 (surface marker of M1 macrophage polarization) and CD206 (surface marker of M2 macrophage polarization) were measured by flow cytometry. The expression levels of signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB) and phosphorylated (p)-NF-κB were detected by Western blot (WB). The migration ability of CAL-27 cells was assessed by scratch assay.Results:Compared with the control group, the expression levels of IL-1βand TNF-α mRNA, and IL-1βand CD86 proteins were up-regulated, whereas those of IL-10 and TGF-β mRNA, and IL-10 and CD206 proteins were down-regulated in the hyperthermia, ferric citrate and hyperthermia + ferric citrate groups, respectively (all P<0.05). In addition, the changes in the hyperthermia+ferric citrate group were significantly larger than those in the hyperthermia and ferric citrate groups (all P<0.05). WB showed that the expression level of STAT3 protein was down-regulated and those of TLR4, NF-κB and p-NF-κB expression were up-regulated in the hyperthermia + ferric citrate group (all P<0.05). Scratch assay showed that the migration ability of CAL-27 cells was inhibited in the hyperthermia, ferric citrate and hyperthermia + ferric citrate groups ( P<0.001), and the changes in the hyperthermia + ferric citrate group were significantly more pronounced than those in the hyperthermia and ferric citrate groups (both P<0.001). Conclusions:Hyperthermia and iron overload can promote the polarization of M1 macrophages and inhibit the polarization of M1 macrophages into M2 macrophages, thereby suppressing the migration of oral squamous cell carcinoma. The mechanism may be related to inhibiting the expression of STAT3 and activating the TLR4/NF-κB signaling pathway.

Objective:To investigate the efficacy of blepharoplasty using the "double-line technique" in addressing upper eyelid skin laxity in middle-aged and elderly patients.Methods:A retrospective analysis was conducted on the clinical data of middle-aged and elderly patients with moderate to severe upper eyelid skin laxity who were admitted to the Plastic Surgery Department of Shaanxi Provincial People’s Hospital between January 2020 and January 2023. The same surgeon performed the "double-line" blepharoplasty to excise excess upper eyelid skin for laxity correction. Patients were positioned flat with their eyes closed; an arcuate line a (5-7 mm above the pupil) and a second line b (aligned with the eyelid margin in an upright position) were marked. The skin between these two lines was excised. Postoperatively, the double eyelid shape, improvement in skin laxity, adverse reactions, and patient satisfaction (categorized as satisfied, basically satisfied, unsatisfied) were assessed.Results:A total of 145 patients were included in the study, comprising 27 males and 118 females. Their ages ranged from 38 to 78 years, with an average age of 52 years. All patients’ incisions healed primarily without complications such as hematoma and infection. During the 3 to 12 months of follow-up after surgery, the double eyelid arcs on both sides appeared symmetrical and natural. The upper eyelid laxity was effectively corrected, obscured vision was improved, crow’s feet were reduced, and the periorbital area was rejuvenated. Patient satisfaction with the surgical results was high, with 112 patients reporting satisfaction, 25 reporting basically satisfied, and 8 reporting dissatisfaction. The overall satisfaction rate reached 94.48%(137/145). Among those dissatisfied, 6 cases cited the double eyelid crease as being too narrow, and 2 cases noted asymmetry in the double eyelid crease.Conclusion:The "double-line method" for blepharoplasty is straightforward to perform and effectively removes excess upper eyelid skin. It is suitable for middle-aged and elderly patients with moderate to severe upper eyelid skin laxity, resulting in favorable surgical outcomes.

Objective:To investigate the efficacy of blepharoplasty using the "double-line technique" in addressing upper eyelid skin laxity in middle-aged and elderly patients.Methods:A retrospective analysis was conducted on the clinical data of middle-aged and elderly patients with moderate to severe upper eyelid skin laxity who were admitted to the Plastic Surgery Department of Shaanxi Provincial People’s Hospital between January 2020 and January 2023. The same surgeon performed the "double-line" blepharoplasty to excise excess upper eyelid skin for laxity correction. Patients were positioned flat with their eyes closed; an arcuate line a (5-7 mm above the pupil) and a second line b (aligned with the eyelid margin in an upright position) were marked. The skin between these two lines was excised. Postoperatively, the double eyelid shape, improvement in skin laxity, adverse reactions, and patient satisfaction (categorized as satisfied, basically satisfied, unsatisfied) were assessed.Results:A total of 145 patients were included in the study, comprising 27 males and 118 females. Their ages ranged from 38 to 78 years, with an average age of 52 years. All patients’ incisions healed primarily without complications such as hematoma and infection. During the 3 to 12 months of follow-up after surgery, the double eyelid arcs on both sides appeared symmetrical and natural. The upper eyelid laxity was effectively corrected, obscured vision was improved, crow’s feet were reduced, and the periorbital area was rejuvenated. Patient satisfaction with the surgical results was high, with 112 patients reporting satisfaction, 25 reporting basically satisfied, and 8 reporting dissatisfaction. The overall satisfaction rate reached 94.48%(137/145). Among those dissatisfied, 6 cases cited the double eyelid crease as being too narrow, and 2 cases noted asymmetry in the double eyelid crease.Conclusion:The "double-line method" for blepharoplasty is straightforward to perform and effectively removes excess upper eyelid skin. It is suitable for middle-aged and elderly patients with moderate to severe upper eyelid skin laxity, resulting in favorable surgical outcomes.

Objective:To observe the effect of hyperthermia combined with iron overload on the regulation of tumor-associated macrophage polarization and the migration ability of CAL-27 cells and explore its mechanism.Methods:Human monocytic leukemia cell THP-1 was induced and polarized into M2 macrophages. M2 tumor-associated macrophages and CAL-27 cells were divided into the control group (no intervention), hyperthermia group (incubated at 42 ℃ for 1 h, and then incubated at 37 ℃ for 24 h), ferric citrate group (added with 2.5 mg/ml ferric citrate, and cultured in an incubator at 37 ℃for 24 h) and hyperthermia + ferric citrate group (added with 2.5 mg/ml ferric citrate for 1 h, cultured in an incubator at 42℃ for 1 h, and then incubated at 37℃ for 24 h). For M2 macrophage groups, the mRNA relative expression levels of surface markers of M1 macrophage polarization including interleukin (IL)-1β, tumor necrosis factor-α(TNF-α), and those of M2 macrophage polarization including IL-10 and transforming growth factor-β(TGF-β) were detected by real-time reverse transcription polymerase chain reaction. The expression levels of IL-1β and IL-10 were detected by ELISA. The expression levels of CD86 (surface marker of M1 macrophage polarization) and CD206 (surface marker of M2 macrophage polarization) were measured by flow cytometry. The expression levels of signal transducer and activator of transcription 3 (STAT3), Toll-like receptor 4 (TLR4), nuclear factor κB (NF-κB) and phosphorylated (p)-NF-κB were detected by Western blot (WB). The migration ability of CAL-27 cells was assessed by scratch assay.Results:Compared with the control group, the expression levels of IL-1βand TNF-α mRNA, and IL-1βand CD86 proteins were up-regulated, whereas those of IL-10 and TGF-β mRNA, and IL-10 and CD206 proteins were down-regulated in the hyperthermia, ferric citrate and hyperthermia + ferric citrate groups, respectively (all P<0.05). In addition, the changes in the hyperthermia+ferric citrate group were significantly larger than those in the hyperthermia and ferric citrate groups (all P<0.05). WB showed that the expression level of STAT3 protein was down-regulated and those of TLR4, NF-κB and p-NF-κB expression were up-regulated in the hyperthermia + ferric citrate group (all P<0.05). Scratch assay showed that the migration ability of CAL-27 cells was inhibited in the hyperthermia, ferric citrate and hyperthermia + ferric citrate groups ( P<0.001), and the changes in the hyperthermia + ferric citrate group were significantly more pronounced than those in the hyperthermia and ferric citrate groups (both P<0.001). Conclusions:Hyperthermia and iron overload can promote the polarization of M1 macrophages and inhibit the polarization of M1 macrophages into M2 macrophages, thereby suppressing the migration of oral squamous cell carcinoma. The mechanism may be related to inhibiting the expression of STAT3 and activating the TLR4/NF-κB signaling pathway.

Objective To investigate the impacts of cinobufotalin combined with docetaxel+cisplatin chemotherapy on the disease control and the level of vascular endothelial growth factor(VEGF)in serum of patients with non-small cell lung cancer.Methods Totally 70 patients with non-small cell lung cancer admitted to Zhejiang Veteran Hospital from March 2019 to March 2022 were included.They were randomly divided into combination group(cinobufotalin combined with docetaxel+cisplatin,n= 35)and control group(docetaxel+cisplatin chemotherapy alone,n=35).The disease control rates of two groups,serum VEGF level,carcinoembryonic antigen(CEA),cy-tokeratin 19(CK19)and carbohydrate antigen 125(CA125)were measured using ELISA;life quality of patients was evaluated with the KPS score;The adverse events between two groups were compared.Results The disease control rate in the combination group(33/35,94.29％)was higher than that that of control group(25/35,71.43％,)(P＜0.05).After treatment,the level of VEGF in both groups of patients decreased,and the KPS score increased(P＜0.05)especially in the combination group(P＜0.05).After treatment,the level of serum CEA,CA125,and CK19 of control group and combination group were obviously lower than those before treatment(P＜0.05),while those in the combination group were even lower(P＜0.05).The incidence of adverse events in the combination group(5/35,14.29％)was lower than that in the control group(13/35,37.14％)(P＜0.05).Conclusions Cinobufotalin combined with docetaxel+cisplatin chemotherapy as a potential new chemotherapy significantly reduces the level of VEGF and tumor biomarkers in serum factor,improves the life quality of patients.The combined therapy is proved tobe safe.

Objective:To investigate the regulation and possible mechanism of hyperthermia (HT) on the ferroptosis of squamous cell carcinoma of the tongue cell line CAL-27.Methods:Half maximal inhibitory concentration (IC 50) of Fer-1, an inhibitor of ferroptosis, was detected by CCK-8 assay and used for subsequent experiments. CAL-27 cells were divided into the HT, control, Fer-1 and HT+ Fer-1 groups according to experimental design. Reactive oxygen species (ROS) levels and iron ion concentration were determined by corresponding detection kits. The p53 and TfR1 mRNA levels were detected by real-time reverse transcription PCR. Cell migration was detected by cell scratch test and cell apoptosis was detected by flow cytometry. Results:HT significantly up-regulated the ROS levels ( P<0.01) and iron ion concentration ( P<0.001), and significantly increased the expression levels of p53 and TfR1 mRNA (both P<0.01). The cell migration ability was decreased ( P<0.001), whereas cell apoptosis rate was increased by HT ( P<0.01). In the HT+Fer-1 group, the ROS levels ( P<0.001), iron ion concentration ( P<0.001), expression levels of p53 and TfR1 mRNA (both P<0.01) were significantly down-regulated, the cell migration ability was recovered ( P<0.01), and cell apoptosis rate was decreased ( P<0.01) compared with those in the HT group, respectively. Conclusions:HT may induce the ferroptosis of CAL-27 cell line, inhibit cell migration ability and promote cell apoptosis by activating the p53/TfR1 pathway.

Objective:To observe the effect of hyperthermia combined with paclitaxel on the proliferation, apoptosis and cycle of human tongue squamous cell carcinoma cell line CAL-27, and to explore the underlying mechanism.Methods:The working concentration of paclitaxel was determined by CCK-8 assay, and the cultured CAL-27 cells were divided into the control, paclitaxel, 42℃ hyperthermia and combined treatment groups. The ability of cell proliferation was detected by colony formation assay, and the cell cycle and apoptosis were determined by flow cytometry. The expression levels of AKT, p-AKT, Bcl-2 and Bax proteins in each group were measured by Western blot.Results:Compared with the control group, the proliferation was significantly inhibited and the apoptosis of CAL-27 cells was significantly promoted in the combined treatment, hyperthermia and paclitaxel groups (all P<0.05), and the anti-proliferation and apoptosis-promoting effect in the combined treatment group was significantly better than those in the hyperthermia and paclitaxel groups (all P<0.05). Western blot showed that hyperthermia combined with paclitaxel could significantly up-regulate the expression level of Bax protein and significantly down-regulate the expression levels of P-AKT and Bcl-2 in CAL-27 cells (all P<0.05). Conclusions:Hyperthermia combined with paclitaxel can play a synergistic role in inhibiting proliferation and promoting apoptosis of tongue squamous cell carcinoma CAL-27 cells. The mechanism may be related to the inhibition of AKT activation and the activation of Bax/Bcl-2 apoptosis signaling pathway.

Metabolic reprogramming is a malignancy hallmark, which refers to the ability of cancer cells to alter metabolic and nutrient acquisition modes in order to support the energy demands for accomplishing the rapid growth, dissemination, metastasis and obtain the "building blocks" needed to maintain cell division. When solid tumors are exposed to low pH, low oxygen and tumor microenvironment with nutrient deficiencies, the hypoxia-inducible factor-1 can be activated, which mediates the remodeling of metabolic patterns in tumor cells, namely, energy is obtained by circulating intracellular components (removing substrates such as proteins and lipid) or by utilizing adaptive metabolic reprogramming (such as glycolysis, autophagy and lipid metabolism, etc.). As a treatment scheme based on local heating of tumors, hyperthermia has a variety of anticancer mechanisms and can be used in combination with radiotherapy, chemotherapy and biological immune therapy. In this review, we briefly discussed the metabolic remodeling model mediated by hypoxia-inducible factor 1 in a hypoxia microenvironment, described the possible regulatory mechanism of hyperthermia on hypoxia-inducible factor-1 and prospected the application of hyperthermia in oral and maxillofacial tumors.

Objective:The investigate the roles and significance of HIF-1α and CYPJ in tongue squamous cell carcinoma cell (TSCC), and further evaluate the regulatory effect of hyperthermia (HT) on HIF-1α and CYPJ in TSCC cells.Methods:Eighty samples of cancer tissues and adjacent normal tissues from TSCC patients were collected. The expression levels of HIF-1α and CYPJ were detected by immunohistochemistry, Western blotting (WB) and fluorescence quantitative PCR, and the relationship between the expression levels of HIF-1α and CYPJ and clinicopathological characteristics was further analyzed. The expression levels of HIF-1α and CYPJ in Cal-27 cells under normoxic and hypoxic conditions for 24 h when combined with HT (42℃), chemotherapy and both were detected by qPCR and WB. Cell migration was detected by cell scratch test and cell apoptosis was measured by flow cytometry.Results:The expression levels of HIF-1α and CYPJ proteins in the tumor tissues of TSCC patients were higher than those in the adjacent normal tissues, which were significantly correlated with tumor size, TNM stage, differentiation degree and lymph node metastasis in TSCC patients (all P<0.05), whereas they were not correlated with gender or age (all P>0.05). The expression levels of HIF-1α and CYPJ in Cal-27 cells were significantly up-regulated in the hypoxic microenvironment (both P<0.05), which were also significantly enhanced by hyperthermia alone (both P<0.05). Compared with hyperthermia or chemotherapy alone, hyperthermia combined with chemotherapy significantly inhibited the expression of HIF-1α and CYPJ, suppressed cell migration and promoted cell apoptosis (all P<0.05). Conclusions:HIF-1α and CYPJ may be potential biomarkers for TSCC tumorigenicity and prognosis. In addition, tumor recurrence after hyperthermia may be due to the role of hyperthermia in triggering HIF-1α expression, which promotes the growth and survival of tumor cells adaptive to hyperthermia treatment by activating the downstream target genes, while hyperthermia combined with chemotherapy may be a promising treatment for TSCC.

With the in-depth study of tumor hyperthermia and tumor immune microenvironment (TIME), the role of hyperthermia in TIME has captivated increasing attention from scholars in recent years. Based upon recent research progress at home and abroad, the effect and mechanism of hyperthermia on several major immune cells and immune-related cytokines in the TIME were reviewed in this article. Comprehensive and deep understanding of the regulation of hyperthermia on the TIME could provide new ideas and methods for tumor treatment.