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.

Background and purpose:Lobaplatin,as a traditional chemotherapeutic drug,is widely used in the treatment of malignant tumor.In recent years,its application in the field of hyperthermic intraperitoneal chemotherapy(HIPEC)has garnered increasing attention.This study evaluated the efficacy and safety of lobaplatin-based HIPEC in advanced abdominal metastatic cancer.Methods:This study collected data of patients with advanced cancers and malignant ascites who treated in the Cancer Center of Shanghai Tenth People's Hospital,Tongji University School of Medicine,from January 2019 to January 2023.We excluded patients who did not meet the inclusion criteria.Short-term efficacy was assessed by changes in ascitic fluid volume,and long-term survival was analyzed using the Kaplan-Meier method.The correlation between CA12-5 levels before and after treatment was evaluated using Pearson correlation analysis.Baseline characteristics and treatment outcomes were described using descriptive statistics,and the changes in CA12-5 levels before and after treatment were compared using significance tests(P＜0.01).Data entry and statistical analyses were conducted using SPSS version 26.0,and survival curves and efficacy plots were generated with GraphPad Prism(10.4.0 version).The study was approved by the Ethics Committee of Shanghai Tenth People's Hospital(Ethics approval number:SHSY-IEC-5.0/24K134/P01).This prospective single-arm study strictly adhered to the guideline of Consolidated Standards of Reporting Trials(CONSORT)checklist.Results:A total of 21 patients were enrolled in this study.The median age of the patients was 61 years(ranging from 31 to 71 years).Among the 21 patients,5(23.8%)achieved complete remission(CR),5(23.8%)achieved partial remission(PR),8(38.1%)had stable disease(SD),and 3(14.3%)experienced disease progression(PD).The overall response rate(ORR)was 47.6%,and the disease control rate(DCR)was 85.7%.Survival analysis revealed a median progression-free survival(PFS)of 12.33 months and a median overall survival(OS)of 16.37 months.Analysis of tumor markers showed a significant negative correlation between efficacy and CA12-5 levels(P＜0.01).Adverse reactions primarily included myelosuppression,hepatic and renal impairment,and nausea and vomiting,with most adverse events being mild to moderate.Conclusion:Lobaplatin-based HIPEC is effective in the treatment of advanced gastrointestinal malignancies with malignant ascites,providing survival benefits and demonstrating good safety.CA12-5 may serve as a valuable predictor of poor prognosis.

Objective:The study retrospectively analyzed the etiology, clinical manifestations, emergency treatment and etiological treatment of a large sample of cases with hypercalcemic crisis.Methods:The clincial data of patients with hypercalcaemia cirisis who were administered in First Medical Center of Chinese PLA General Hospital from January 2009 to July 2022 were analyzed, inculding the general data, clinical manifestations, etiology, photographic examination, emergency treatment, etiological treatment, serological examination before and after treatment, pathological immunohistochemical findings and prognosis.Results:A total of 143 hypercalcaemia crisis patients(84 males and 59 females) with a mean age of 53.51±16.60 were enrolled. The most common disease was hyperparathyroidism(62/143), followed by solid malignancy(57/143) and multiple myeloma(12/143). Patients presented with digestive system symptoms at 76.91%, followed by neurological symptoms at 63.60%, urinary system symptoms at 58.76%, musculoskeletal symptoms at 55.23%, and cardiovascular system symptoms at 32.91%. After emergency calcium-lowering treatment, the remission rate of hypercalcemic crisis in 143 patients was 100%(143/143), and after etiological treatment, the remission rate of hypercalcemia was 85.31%(122/143).Conclusion:Early identification, emergency treatment and etiology treatment of hypercalcaemia crisis are essential. Effective treament with comprehensive calcium reduction can quickly relieve clinical symptoms and create opportunities for treatment for the cause. Targeted etiological interventions can lead to the correction or long-term remission of hypercalcemia.

Background and purpose:Lobaplatin,as a traditional chemotherapeutic drug,is widely used in the treatment of malignant tumor.In recent years,its application in the field of hyperthermic intraperitoneal chemotherapy(HIPEC)has garnered increasing attention.This study evaluated the efficacy and safety of lobaplatin-based HIPEC in advanced abdominal metastatic cancer.Methods:This study collected data of patients with advanced cancers and malignant ascites who treated in the Cancer Center of Shanghai Tenth People's Hospital,Tongji University School of Medicine,from January 2019 to January 2023.We excluded patients who did not meet the inclusion criteria.Short-term efficacy was assessed by changes in ascitic fluid volume,and long-term survival was analyzed using the Kaplan-Meier method.The correlation between CA12-5 levels before and after treatment was evaluated using Pearson correlation analysis.Baseline characteristics and treatment outcomes were described using descriptive statistics,and the changes in CA12-5 levels before and after treatment were compared using significance tests(P＜0.01).Data entry and statistical analyses were conducted using SPSS version 26.0,and survival curves and efficacy plots were generated with GraphPad Prism(10.4.0 version).The study was approved by the Ethics Committee of Shanghai Tenth People's Hospital(Ethics approval number:SHSY-IEC-5.0/24K134/P01).This prospective single-arm study strictly adhered to the guideline of Consolidated Standards of Reporting Trials(CONSORT)checklist.Results:A total of 21 patients were enrolled in this study.The median age of the patients was 61 years(ranging from 31 to 71 years).Among the 21 patients,5(23.8%)achieved complete remission(CR),5(23.8%)achieved partial remission(PR),8(38.1%)had stable disease(SD),and 3(14.3%)experienced disease progression(PD).The overall response rate(ORR)was 47.6%,and the disease control rate(DCR)was 85.7%.Survival analysis revealed a median progression-free survival(PFS)of 12.33 months and a median overall survival(OS)of 16.37 months.Analysis of tumor markers showed a significant negative correlation between efficacy and CA12-5 levels(P＜0.01).Adverse reactions primarily included myelosuppression,hepatic and renal impairment,and nausea and vomiting,with most adverse events being mild to moderate.Conclusion:Lobaplatin-based HIPEC is effective in the treatment of advanced gastrointestinal malignancies with malignant ascites,providing survival benefits and demonstrating good safety.CA12-5 may serve as a valuable predictor of poor prognosis.

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.

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.

Objective:To evaluate the effects of different high pressure on the behavior and physiological indexes of hypothalamic-pituitary-adrenal axis (HPA) in rats, so as to provide a theoretical basis for the stress protection in decompression process.Methods:The rats were divided into four groups according to the random number table method and were separately exposed to air pressure of 0 kPa, 170 kPa, 350 kPa and 700 kPa for 60 mins and given decompression for 45 mins. The behaviors of rats were analyzed by animal open field method and video analysis system of the open field immediately after they were taken from the air pressure chamber, and the contents of adrenocorticotrophic hormone (ACTH) and glucocorticoid (GC) in the serum were measured by the enzyme-linked immunosorbent assay (ELISA).Results:Under different pressure, compared with the 0 kPa group, the total distance traveled by the rats ( F=3.354), the average traveling speed ( F=3.358), the number of standing times and standing time duration ( F=3.739; F=33.332) of the rats in the other 3 groups decreased significantly. The distance traveled in the corner of the 175 kPa group and the 700 kPa group decreased significantly ( F=3.532), while their cleaning time significantly increased ( F=4.581). The standing times and the cleaning time of the 350 kPa group were significantly less than those of the 700 kPa group. All the differences above showed statistical significance ( P<0.05). After exposure to different pressure, the ACTH level of the 175 kPa group was higher than those of the 0 kPa group and the 350 kPa group ( F=5.309) with statistical significance ( P<0.05). The GC level of the 0 kPa group was significantly lower than those of the other 3 groups, and the GC level of the 700 kPa group was especially lower than that of the 175 kPa group, with statistically significant difference ( P<0.05). Conclusion:High pressure exposure at all levels can cause definite stress reaction, and the stress reactions do not correlate with the pressure gradient. Within the physiological bearable range of stress under high pressure, the individual stress level may present a certain steady state. The individual′s behavioral response, ACTH and GC levels may be used as indicators for stress monitoring.

Objective:To evaluate the effects of different high pressure on the behavior and physiological indexes of hypothalamic-pituitary-adrenal axis (HPA) in rats, so as to provide a theoretical basis for the stress protection in decompression process.Methods:The rats were divided into four groups according to the random number table method and were separately exposed to air pressure of 0 kPa, 170 kPa, 350 kPa and 700 kPa for 60 mins and given decompression for 45 mins. The behaviors of rats were analyzed by animal open field method and video analysis system of the open field immediately after they were taken from the air pressure chamber, and the contents of adrenocorticotrophic hormone (ACTH) and glucocorticoid (GC) in the serum were measured by the enzyme-linked immunosorbent assay (ELISA).Results:Under different pressure, compared with the 0 kPa group, the total distance traveled by the rats ( F=3.354), the average traveling speed ( F=3.358), the number of standing times and standing time duration ( F=3.739; F=33.332) of the rats in the other 3 groups decreased significantly. The distance traveled in the corner of the 175 kPa group and the 700 kPa group decreased significantly ( F=3.532), while their cleaning time significantly increased ( F=4.581). The standing times and the cleaning time of the 350 kPa group were significantly less than those of the 700 kPa group. All the differences above showed statistical significance ( P<0.05). After exposure to different pressure, the ACTH level of the 175 kPa group was higher than those of the 0 kPa group and the 350 kPa group ( F=5.309) with statistical significance ( P<0.05). The GC level of the 0 kPa group was significantly lower than those of the other 3 groups, and the GC level of the 700 kPa group was especially lower than that of the 175 kPa group, with statistically significant difference ( P<0.05). Conclusion:High pressure exposure at all levels can cause definite stress reaction, and the stress reactions do not correlate with the pressure gradient. Within the physiological bearable range of stress under high pressure, the individual stress level may present a certain steady state. The individual′s behavioral response, ACTH and GC levels may be used as indicators for stress monitoring.