Background and purpose:Melanoma is a highly invasive malignant tumor originating from melanocytes,which poses a great threat to human life and health around the world,and its morbidity and mortality have been rising continuously in recent years.Telomerase and autophagy play crucial roles in cell proliferation,survival and stress response.Telomerase maintains the replication ability of cells by prolonging telomeres at the ends of chromosomes,and autophagy,as a self-degradation mechanism of cells,can not only help cells remove damaged components to promote survival,but also induce cell death under certain conditions.In the tumor environment,they are often abnormally activated or out of balance,and participate in the occurrence and development of many cancers,including melanoma.This study investigated the roles of telomerase and autophagy in melanoma progression and evaluated the potential synergistic therapeutic effects of combined application of telomerase inhibitor BIBR1532 and autophagy inhibitor chloroquine(CQ)in melanoma treatment.Methods:Malignant melanoma cells A375 were treated with telomerase inhibitor BIBR1532.The cell viability was assessed using the cell counting kit-8(CCK-8)assay,and the cell apoptosis was detected using the Annexin Ⅴ/propidium iodide(PI)double staining method.Additionally,the expressions of autophagy-related proteins LC3-Ⅱand p62 were detected by Western blot,and the changes in autophagy flux were observed using dual-tagged adenovirus transfection technology.Based on these studies,BIBR1532 and the autophagy inhibitor CQ were further applied in combination to analyze cell proliferation,apoptotic rate,changes in mitochondrial membrane potential,and cell cycle distribution,and the cloning formation experiment was used to verify the cell's proliferative capacity,thereby comprehensively evaluating the efficacy of this combined treatment strategy.Results:Telomerase inhibitor BIBR1532 at a concentration of 50 μmol/L significantly inhibited the growth of malignant melanoma cells A375 and induced apoptosis.At the same concentration,BIBR1532 upregulated the expression of the autophagy-related protein LC3-Ⅱ in A375 cells,while downregulating the expression of p62 protein.By transducing A375 cells with a dual-tagged adenovirus,it was observed that autophagy flux was significantly enhanced after treatment with BIBR1532.Furthermore,the combined application of BIBR1532(50 μmol/L)and the autophagy inhibitor CQ(20 μmol/L)significantly promoted the death of A375 cells,induced apoptosis and destruction of mitochondrial membrane potential,caused cell cycle arrest at the G2/M phase,and significantly inhibited the cell's clonogenic ability.Conclusion:Telomerase inhibitor BIBR1532 not only inhibits the proliferation of malignant melanoma cells but also activates the autophagy process in these cells,and inhibition of the autophagy response by autophagy inhibitor CQ can enhance the sensitivity of malignant melanoma cells to telomerase inhibitor BIBR1532.

Background and purpose:Melanoma is a highly invasive malignant tumor originating from melanocytes,which poses a great threat to human life and health around the world,and its morbidity and mortality have been rising continuously in recent years.Telomerase and autophagy play crucial roles in cell proliferation,survival and stress response.Telomerase maintains the replication ability of cells by prolonging telomeres at the ends of chromosomes,and autophagy,as a self-degradation mechanism of cells,can not only help cells remove damaged components to promote survival,but also induce cell death under certain conditions.In the tumor environment,they are often abnormally activated or out of balance,and participate in the occurrence and development of many cancers,including melanoma.This study investigated the roles of telomerase and autophagy in melanoma progression and evaluated the potential synergistic therapeutic effects of combined application of telomerase inhibitor BIBR1532 and autophagy inhibitor chloroquine(CQ)in melanoma treatment.Methods:Malignant melanoma cells A375 were treated with telomerase inhibitor BIBR1532.The cell viability was assessed using the cell counting kit-8(CCK-8)assay,and the cell apoptosis was detected using the Annexin Ⅴ/propidium iodide(PI)double staining method.Additionally,the expressions of autophagy-related proteins LC3-Ⅱand p62 were detected by Western blot,and the changes in autophagy flux were observed using dual-tagged adenovirus transfection technology.Based on these studies,BIBR1532 and the autophagy inhibitor CQ were further applied in combination to analyze cell proliferation,apoptotic rate,changes in mitochondrial membrane potential,and cell cycle distribution,and the cloning formation experiment was used to verify the cell's proliferative capacity,thereby comprehensively evaluating the efficacy of this combined treatment strategy.Results:Telomerase inhibitor BIBR1532 at a concentration of 50 μmol/L significantly inhibited the growth of malignant melanoma cells A375 and induced apoptosis.At the same concentration,BIBR1532 upregulated the expression of the autophagy-related protein LC3-Ⅱ in A375 cells,while downregulating the expression of p62 protein.By transducing A375 cells with a dual-tagged adenovirus,it was observed that autophagy flux was significantly enhanced after treatment with BIBR1532.Furthermore,the combined application of BIBR1532(50 μmol/L)and the autophagy inhibitor CQ(20 μmol/L)significantly promoted the death of A375 cells,induced apoptosis and destruction of mitochondrial membrane potential,caused cell cycle arrest at the G2/M phase,and significantly inhibited the cell's clonogenic ability.Conclusion:Telomerase inhibitor BIBR1532 not only inhibits the proliferation of malignant melanoma cells but also activates the autophagy process in these cells,and inhibition of the autophagy response by autophagy inhibitor CQ can enhance the sensitivity of malignant melanoma cells to telomerase inhibitor BIBR1532.

Objective To investigate the expression levels and clinical significance of cold-inducible RNA-binding protein(CIRBP)and triggering receptor expressed on myeloid cells-1(TREM-1)in peripheral blood of patients with chronic obstructive pulmonary disease(COPD).Methods Forty hospitalized COPD patients from October 2022 to June 2023 were selected as the acute exacerbation group.After treatment,they entered the stable phase and were included in the stable phase group.During the same period,40 healthy individuals underwent physical exami-nations as the control group.General information from each group was collected,peripheral blood was collected,and the levels of CIRBP and TREM-1 in plasma were measured using enzyme-linked immunosorbent assay(ELISA).The relative expression levels of CIRBP mRNA and TREM-1 mRNA were measured using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),and lung function was measured in COPD patients and healthy individuals.Results The expression levels of CIRBP and TREM-1 in peripheral blood during the acute exacerba-tion of COPD were higher than those in the stable phase group,and the differences were statistically significant(all P＜0.001);The expression levels of CIRBP and TREM-1 in peripheral blood during acute exacerbation and stable phases were higher than those in the control group,and the differences were statistically significant(all P＜0.001);In both acute exacerbation and stable phases,CIRBP levels were positively correlated with TREM-1 levels;The levels of CIRBP and TREM-1 during acute exacerbation were positively correlated with white blood cell count,neutrophil percentage,neutrophil to lymphocyte ratio;The stable CIRBP and TREM-1 levels were negatively correla-ted with the percentage of forced expiratory volume at 1 second to the expected value,and the ratio of forced expira-tory volume to forced vital capacity at 1 second.They were positively correlated with white blood cell count,neutro-phil percentage,and neutrophil/lymphocyte ratio.Conclusion The expression levels of CIRBP and TREM-1 are el-evated in the peripheral blood of COPD patients.The expression of CIRBP is correlated with TREM-1 expression,and is associated with clinical indicators such as lung function,white blood cell count,neutrophil percentage,neutro-phil to lymphocyte ratio,monocyte to high-density lipoprotein ratio,etc.,suggesting that CIRBP and TREM-1 may jointly participate in the occurrence and development of COPD.

Objective To investigate the effect of different doses of rocuronium on the monitoring of recurrent laryngeal nerve during endoscopic thyroidectomy.Methods A total of 116 patients undergoing endoscopic thyroidectomy through areolar approach were selected from October 2021 to October 2022,30 males and 86 females,aged 18-64 years,BMI 18.5-30.0 kg/m2,ASA physical status Ⅰ or Ⅱ.All the patients were divided into three groups by random number table method:rocuronium 0.30 mg/kg group(group R1,n=39),rocuronium 0.45 mg/kg group(group R2,n=39),and rocuronium 0.60 mg/kg group(group R3,n=38).After induction of anesthesia,groups R1,R2,and R3 were injected intrave-nously with rocuronium 0.30,0.45,and 0.60 mg/kg,respectively.When the TOF value was 0,the nerve monitoring tracheal catheter was inserted,and the muscle relaxation was monitored throughout the operation.No muscle relaxants were added before the end of the nerve monitoring.The time and amplitude of recurrent laryngeal nerve electromyography(EMG)from intravenous rocuronium to the first occurrence were recorded.The time of intubation and quality of tracheal intubation(Cooper's score),intraoperative special conditions(hypotension,hypertension,bradycardia,tachycardia,intraoperative movement,etc.),postoperative throat pain,hoarseness,and muscle pain were recorded.Results There was no significant difference in the time of first occurrence of recurrent laryngeal nerve EMG among the three groups.Compared with group R1,the recurrent laryngeal nerve EMG amplitude in groups R2 and R3 was significantly decreased for the first occurrence(P＜0.05).Compared with group R1,the time of intubation in groups R2 and R3 was signifi-cantly shortened(P＜0.05).Compared with group R2,the time of intubation in group R3 was significantly shortened(P＜0.05).Compared with group R1,the quality of tracheal intubation in groups R2 and R3 was significantly higher(P＜0.05).Compared with group R1,the incidence of intraoperative and postop-erative laryngeal pain in groups R2 and R3 was significantly lower(P＜0.05).Conclusion During endo-scopic thyroidectomy,compared with rocuronium 0.30 mg/kg,rocuronium 0.45 and 0.60 mg/kg can not only provide good conditions for tracheal intubation,but also monitor recurrent laryngeal nerve signals,and rocuronium 0.60 mg/kg can be intubated for a shorter time.

OBJECTIVES: Peripheral whole blood cell counts have been used as prognostic indicators for various cancers, but their predictive value in nasopharyngeal carcinoma remain unclear. This study aims to evaluate the prognostic significance of the pretreatment hemoglobin×lymphocyte/monocyte ratio (HLMR) in non-recurrent, non-metastatic NPC patients undergoing definitive radiotherapy. METHODS: Clinical and follow-up data from 805 NPC patients who completed definitive radiotherapy or chemoradiotherapy were retrospectively analyzed. Pretreatment hemoglobin, lymphocyte count, and monocyte count were collected to calculate HLMR. Receiver operating characteristic (ROC) curves were used to determine the optimal cut-off value of HLMR. Patients were then classified into high and low HLMR groups. The association between HLMR and clinicopathological characteristic was assessed using chi-square tests. Independent prognostic factors for overall survival (OS) and progression-free survival (PFS) were identified using Cox proportional hazards models. A nomogram was constructed based on the independent predictors to estimate patient survival rates, and internal validation was performed using a validation cohort. RESULTS: The ROC curve identified 605.5 as the optimal HLMR cut-off value for predicting 5-year survival. Multivariate Cox regression analysis revealed that T stage (HR=1.886, 95% CI 1.331 to 2.673, P<0.001), N stage (HR=2.021, 95% CI 1.267 to 3.225, P=0.003), Eastern Cooperative Oncology Group (ECOG) score (HR=3.991, 95% CI 1.257 to 12.677, P=0.019), concurrent chemoradiotherapy regimen (HR=0.338, 95% CI 0.156 to 0.731, P=0.006), and HLMR (HR=0.648, 95% CI 0.460 to 0.912, P=0.013) were independent prognostic factors for OS. A nomogram including T stage, N stage, and HLMR in the training cohort was constructed to predict 3-, 5-, and 7-year OS, with a C-index of 0.713. The area under the curves for predicting 3-, 5-, and 7-year OS were 0.744, 0.665, and 0.682, respectively. Calibration curves showed good agreement between predicted and observed survival rates. The above results were further confirmed in the validation cohort. CONCLUSIONS Pretreatment HLMR may serve as a promising prognostic biomarker for patients with nasopharyngeal carcinoma.
Humans ; Nasopharyngeal Carcinoma/mortality* ; Prognosis ; Hemoglobins/analysis* ; Nasopharyngeal Neoplasms/pathology* ; Monocytes/cytology* ; Female ; Male ; Retrospective Studies ; Middle Aged ; Adult ; Aged ; Nomograms ; Chemoradiotherapy ; ROC Curve

The consumption of a high-fat diet(HFD)has been linked to osteoporosis and an increased risk of fragility fractures.However,the specific mechanisms of HFD-induced osteoporosis are not fully understood.Our study shows that exposure to an HFD induces premature senescence in bone marrow mesenchymal stem cells(BMSCs),diminishing their proliferation and osteogenic capability,and thereby contributes to osteoporosis.Transcriptomic and chromatin accessibility analyses revealed the decreased chromatin accessibility of vitamin D receptor(VDR)-binding sequences and decreased VDR signaling in BMSCs from HFD-fed mice,suggesting that VDR is a key regulator of BMSC senescence.Notably,the administration of a VDR activator to HFD-fed mice rescued BMSC senescence and significantly improved osteogenesis,bone mass,and other bone parameters.Mechanistically,VDR activation reduced BMSC senescence by decreasing intracellular reactive oxygen species(ROS)levels and preserving mitochondrial function.Our findings not only elucidate the mechanisms by which an HFD induces BMSC senescence and associated osteoporosis but also offer new insights into treating HFD-induced osteoporosis by targeting the VDR-superoxide dismutase 2(SOD2)-ROS axis.

The consumption of a high-fat diet(HFD)has been linked to osteoporosis and an increased risk of fragility fractures.However,the specific mechanisms of HFD-induced osteoporosis are not fully understood.Our study shows that exposure to an HFD induces premature senescence in bone marrow mesenchymal stem cells(BMSCs),diminishing their proliferation and osteogenic capability,and thereby contributes to osteoporosis.Transcriptomic and chromatin accessibility analyses revealed the decreased chromatin accessibility of vitamin D receptor(VDR)-binding sequences and decreased VDR signaling in BMSCs from HFD-fed mice,suggesting that VDR is a key regulator of BMSC senescence.Notably,the administration of a VDR activator to HFD-fed mice rescued BMSC senescence and significantly improved osteogenesis,bone mass,and other bone parameters.Mechanistically,VDR activation reduced BMSC senescence by decreasing intracellular reactive oxygen species(ROS)levels and preserving mitochondrial function.Our findings not only elucidate the mechanisms by which an HFD induces BMSC senescence and associated osteoporosis but also offer new insights into treating HFD-induced osteoporosis by targeting the VDR-superoxide dismutase 2(SOD2)-ROS axis.

Humans ; Hematopoietic Stem Cell Transplantation ; Multiple Myeloma/therapy* ; Transplantation, Autologous ; Retrospective Studies ; Combined Modality Therapy ; Stem Cell Transplantation/adverse effects*

Objective:To analyze the clinical characteristics of fulminant type 1 diabetes (FT1DM) in China.Methods:Clinical data of 279 cases related to FT1DM in Chinese Database from January 2005 to December 2018 were collected, and other 20 patients from our hospital were included in the present study.Results:(1) There has been a progressive increasing in the number of reported cases every year in China, and the number in the southern region were significantly more than that in the northern region. (2) The median age of the onset of FT1DM patients in China was 32.5 years old, without significant gender difference. Moreover, 36.5% (54/148) of the female patients caught the disease during their prenatal period, most of them were onset in the second or third trimesters of pregnancy and 2 weeks after delivery (37/40), and the prognosis of the fetus was extremely poor. (3) Compared with new-onset type 1 diabetes, FT1DM patients were younger, and with higher blood glucose [(39.7±15.3) vs (21.2 ± 9.9) mmol/L], higher serum creatinine [(188.4±115.9) vs (51.8 ±23.1) μmol/L], and higher amylase levels [245.5 (26.0-5 137.0) vs 54.7 (14.0-404.9) U/L]. FT1DM patients were with more severe acidosis, and lower HbA 1C level [(6.6 ±0.8)% vs (12.9 ± 2.5)%, P<0.01]. (4) FT1DM patients may combine with multiple organ dysfunction or severe metabolic disorders, electrolyte disorders, as well as liver and kidney dysfunctions, and elevation of amylase and muscle enzymes. Conclusion:FT1DM are with some clinical characteristics different from classic new-onset type 1 diabetes, including adult-onset, frequent in the southern China. Pregnancy may be a predisposing factor for female patients. Significant metabolic disorders and multiple organ involvements are common in the patients with FT1DM.

Objective:To evaluate the changes of cortisol and sIgA in serum and saliva of rats under different stresses and pressures, so as to provide a basis for screening non-invasive stress monitoring indicators in high-pressure working environment.Methods:A total of 54 rats were divided into six groups exposed to different air pressures of 0 kPa, 175 kPa, 350 kPa, 500 kPa, 600 kPa, and 700 kPa, respectively. According to different stress conditions, another 36 rats were divided into physiological stress group (PSSG), psychological stress group (PCSG), physiological and psychological stress group (PPSG), and blank group (BG), with 9 rats in each group. At the end of the experiment, serum sample was collected and supernatant was taken from the sublingual gland tissue homogenate. Cortisol and sIgA levels in saliva and serum were determined by radioimmunoassay and enzyme-linked immunosorbent assay (ELISA).Results:The concentrations of SIgA in group 500 kPa, 600 kPa, and 700 kPa were significantly different ( F=10.961, P<0.001; F=4.693, P=0.001; F=4.353, P=0.003). The serum cortisol levels in group 175 kPa, 350 kPa, and 600 kPa were significantly higher than that in group 0 kPa, while the serum cortisol level in group 350 kPa was significantly higher than those in group 500 kPa and 700 kPa. The cortisol concentrations of sublingual gland tissues in group 350 kPa and 700 kPa were significantly higher than those in group 0 kPa, 500 kPa, and 600 kPa. Under different stress conditions, serum sIgA concentrations in PCSG, PSSG, and PPSG were significantly lower than that in BG ( F=4.852, P=0.007; F=4.918, P=0.007; F=3.967, P=0.017). The levels of serum and sublingual cortisol in PCSG and PPSG were significantly higher than those in BG, while the levels of serum and sublingual cortisol in PSSG were significantly lower than those in PCSG. Relevant research results showed that the changes of salivary gland cortisol and serum cortisol were positively correlated under different stress conditions ( r=0.609, P<0.01). Conclusion:Cortisol as a stress monitoring indicator has good effectiveness, and using saliva as an indicator to monitor stress under high pressure environment has a certain degree of feasibility.