Objective To investigate the molecular mechanism by which high mobility group box 1 (HMGB1) mediates renal tubular epithelial cell injury in a high-calcium environment. Methods A CaCl_2 concentration gradient (0, 0.5, 1, 2 mg/mL) treatment system was established based on human renal tubular epithelial cells (HK-2 cells). The calcium ioninduced cell injury effects were evaluated with CCK-8 assay, lactate dehydrogenase (LDH) detection, and DAPI staining. The optimal CaCl_2 concentration was selected for subsequent experiments. The secretion level of HMGB1 in the cell supernatant was detected with ELISA, and the expressions of autophagy markers (LC3-Ⅱ/LC3-Ⅰ, p62) and mTOR pathway-related proteins were detected with Western blot. Lentiviral-mediated HMGB1 silencing (shHMGB1) or pretreatment with ethyl pyruvate (EP), a specific inhibitor of HMGB1, was used on HK-2 cells to verify the role of HMGB1 in cell injury. Additionally, the autophagy agonist rapamycin was added to further clarify the role of the mammalian target of rapamycin (mTOR) signaling axis. Results Different concentrations of CaCl_2 promoted the release of HMGB1, activated autophagy, induced renal tubular epithelial cell injury, increased LDH release, and exacerbated nuclear damage. The optimal concentration was 1 mg/mL in the follow-up experiment. Targeted inhibition of HMGB1 significantly alleviated CaCl_2-induced autophagic response and cell injury. The underlying mechanism might involve the inhibition of HMGB1 expression leading to the blockage of the downstream mTOR signaling pathway, thereby preventing excessive autophagy activation.Conclusion The high-calcium environment activates autophagy through the HMGB1/mTOR signaling axis, which may be one of the important mechanisms by which hypercalciuria promotes renal tubular epithelial cell injury.

Objective:To investigate the function and mechanism of exosomes in bladder cancer cell growth and metastasis regulating by NETs.Methods:The exosomes secreted by SV-HUC-1,MGH-U3 and 253J cells were extracted.PBS and the above exo-somes incubated with PMN,recorded as PBS group,SV-HUC-1 exo group,MGH-U3 exo group,253J exo group.The generation of NETs was detected by Sytox green staining,and the expressions of NETs marker proteins CIT-H3,NE and MPO were detected by Western blot.MGH-U3 and 253J cells were divided into Control group,PMN group,MGH-U3 exo group,253J exo group MGH-U3 exo+PMN group,253J exo+PMN group.The PMN culture supernatant of MGH-U3 exo group and 253J exo group were added with 1.5 U/ml DNAse,after incubation for 30 min,the collected mixture to co-incubate with MGH-U3 and 253J cells,recorded as MGH-U3 exo+PMN+DNAse group and 253J exo+PMN+DNAse group.The proliferation ability of MGH-U3 and 253J cells were detected by CCK8,the migration and invasion of cells were detected by Transwell,and the phosphorylation levels of PI3K and AKT were detected by Western blot.Results:Compared with PBS group,the formation of NETs and the expressions of CIT-H3,NE and MPO in MGH-U3 exo group and 253J exo group increased significantly(P<0.001).There was no significant change of the above indexes in SV-HUC-1 exo group.Compared with Control group,MGH-U3 exo+PMN group and 253J exo+PMN group had significantly increased cell prolifer-ation,migration and invasion ability(P<0.05),the phosphorylation levels of PI3K and AKT increased significantly(P<0.001).Com-pared with MGH-U3 exo+PMN group,MGH-U3 exo+PMN+DNAse group had significantly decreased cell proliferation,migration and invasion ability(P<0.05),the phosphorylation levels of PI3K and AKT decreased significantly(P<0.001).Compared with 293J exo+PMN group,293J exo+PMN+DNAse group had significantly decreased cell proliferation,migration and invasion ability,the phosphor-ylation levels of PI3K and AKT decreased significantly(P<0.001).Conclusion:The exosomes of bladder cancer cells can promote the proliferation,migration and invasion of tumor cells by inducing the generation of NETs.

Objective:To investigate the function and mechanism of exosomes in bladder cancer cell growth and metastasis regulating by NETs.Methods:The exosomes secreted by SV-HUC-1,MGH-U3 and 253J cells were extracted.PBS and the above exo-somes incubated with PMN,recorded as PBS group,SV-HUC-1 exo group,MGH-U3 exo group,253J exo group.The generation of NETs was detected by Sytox green staining,and the expressions of NETs marker proteins CIT-H3,NE and MPO were detected by Western blot.MGH-U3 and 253J cells were divided into Control group,PMN group,MGH-U3 exo group,253J exo group MGH-U3 exo+PMN group,253J exo+PMN group.The PMN culture supernatant of MGH-U3 exo group and 253J exo group were added with 1.5 U/ml DNAse,after incubation for 30 min,the collected mixture to co-incubate with MGH-U3 and 253J cells,recorded as MGH-U3 exo+PMN+DNAse group and 253J exo+PMN+DNAse group.The proliferation ability of MGH-U3 and 253J cells were detected by CCK8,the migration and invasion of cells were detected by Transwell,and the phosphorylation levels of PI3K and AKT were detected by Western blot.Results:Compared with PBS group,the formation of NETs and the expressions of CIT-H3,NE and MPO in MGH-U3 exo group and 253J exo group increased significantly(P<0.001).There was no significant change of the above indexes in SV-HUC-1 exo group.Compared with Control group,MGH-U3 exo+PMN group and 253J exo+PMN group had significantly increased cell prolifer-ation,migration and invasion ability(P<0.05),the phosphorylation levels of PI3K and AKT increased significantly(P<0.001).Com-pared with MGH-U3 exo+PMN group,MGH-U3 exo+PMN+DNAse group had significantly decreased cell proliferation,migration and invasion ability(P<0.05),the phosphorylation levels of PI3K and AKT decreased significantly(P<0.001).Compared with 293J exo+PMN group,293J exo+PMN+DNAse group had significantly decreased cell proliferation,migration and invasion ability,the phosphor-ylation levels of PI3K and AKT decreased significantly(P<0.001).Conclusion:The exosomes of bladder cancer cells can promote the proliferation,migration and invasion of tumor cells by inducing the generation of NETs.

Objective:To study the correlation between myopia growth and physical development of primary school student in Shihezi area of Xinjiang Uyghur Autonomous Region by measuring the ocular biometry and physical parameters of the eye, and to provide an objective basis for the development of myopia prevention and control.Methods:A cross-sectional study design was used. The Grade 1-6 students from 33 schools in Shihezi area were randomly selected in October 2023. The children were classified according to age into the low age group (6-9 years) and the high age group (10-13 years).The children′s height, weight, axial length (AL), maximum keratometry (steepest keratometry, K1), and minimum keratometry (flattest keratometry, K1) were measured. The body mass index (BMI), keratometric astigmatism (ΔK), mean keratometry (K), corneal radius (CR) and AL/CR were calculated. Correlation analyses were performed between ocular biometry and physical parameters. LASSO regression analysis was used to screen the best variables, and linear regression models were constructed after adjusting for confounding factors of age and sex.Results:A total of 3 986 children aged 6-13 years were included in this study, including 2 094 boys (52.53%) and 1 892 girls (47.47%); 2 162 in the lower age group and 1 824 in the upper age group. Girls had higher K, ΔK and AL/CR values than boys (43.38 D vs 42.7 D, t=-7.84, P<0.01; 1.1 D vs 1.01 D, t=-2.14, P=0.03; 2.97 vs 2.96, t=3.32, P=0.02). Boys had higher AL, height, weight and BMI than girls (23.47 mm vs 23.02 mm, t=13.77, P<0.01; 1.34 m vs 1.33 m, t=3.02, P=0.03; 32 kg vs 29.5 kg, t=7.82, P<0.01; 17.4 vs 16.52, t=10.36, P<0.01). LASSO regression was used to screen the 3 physical parameter variables (height, weight, and BMI) to determine that height was the best physical parameter variable for AL/CR. After adjusting for confounders, it was shown that in the low age group (6-9 years), there was a significant positive correlation between children′s height and AL/CR ( b=0.174, 95% CI: 0.106 to 0.241, P<0.001), whereas in the high age group (10-13 years), there was no correlation between children′s height and AL/CR ( b=0.086, 95% CI:-0.038 to 0.21, P=0.174). Conclusion:There was a positive correlation between height and myopia growth in the lower age group of children in Shihezi area, Xinjiang Uyghur Autonomous Region; there was no correlation between height and myopia growth in the higher age group.

Objective:To study the correlation between myopia growth and physical development of primary school student in Shihezi area of Xinjiang Uyghur Autonomous Region by measuring the ocular biometry and physical parameters of the eye, and to provide an objective basis for the development of myopia prevention and control.Methods:A cross-sectional study design was used. The Grade 1-6 students from 33 schools in Shihezi area were randomly selected in October 2023. The children were classified according to age into the low age group (6-9 years) and the high age group (10-13 years).The children′s height, weight, axial length (AL), maximum keratometry (steepest keratometry, K1), and minimum keratometry (flattest keratometry, K1) were measured. The body mass index (BMI), keratometric astigmatism (ΔK), mean keratometry (K), corneal radius (CR) and AL/CR were calculated. Correlation analyses were performed between ocular biometry and physical parameters. LASSO regression analysis was used to screen the best variables, and linear regression models were constructed after adjusting for confounding factors of age and sex.Results:A total of 3 986 children aged 6-13 years were included in this study, including 2 094 boys (52.53%) and 1 892 girls (47.47%); 2 162 in the lower age group and 1 824 in the upper age group. Girls had higher K, ΔK and AL/CR values than boys (43.38 D vs 42.7 D, t=-7.84, P<0.01; 1.1 D vs 1.01 D, t=-2.14, P=0.03; 2.97 vs 2.96, t=3.32, P=0.02). Boys had higher AL, height, weight and BMI than girls (23.47 mm vs 23.02 mm, t=13.77, P<0.01; 1.34 m vs 1.33 m, t=3.02, P=0.03; 32 kg vs 29.5 kg, t=7.82, P<0.01; 17.4 vs 16.52, t=10.36, P<0.01). LASSO regression was used to screen the 3 physical parameter variables (height, weight, and BMI) to determine that height was the best physical parameter variable for AL/CR. After adjusting for confounders, it was shown that in the low age group (6-9 years), there was a significant positive correlation between children′s height and AL/CR ( b=0.174, 95% CI: 0.106 to 0.241, P<0.001), whereas in the high age group (10-13 years), there was no correlation between children′s height and AL/CR ( b=0.086, 95% CI:-0.038 to 0.21, P=0.174). Conclusion:There was a positive correlation between height and myopia growth in the lower age group of children in Shihezi area, Xinjiang Uyghur Autonomous Region; there was no correlation between height and myopia growth in the higher age group.

Objective: To examine the correlation between frailty and lifestyle factors among middle-aged and elderly populations, so as to provide insights into the management of frailty among middle-aged and elderly populations. Methods : Middle-aged and elderly residents at ages of 45 ot 69 years were recruited using the convenient sampling method from seven townships in Changxing County of Zhejiang Province from 2019 to 2020. The demographic characteristics and lifestyle factors were collected using questionnaires, and the frailty was measured using the Chinese version of Tilburg Frailty Indicator ( TFI ). Factors affecting frailty were identified among middle-aged and elderly populations using the multivariable logistic regression model. Results: A total of 7 170 residents were surveyed, including 2 780 males ( 38.77% ) and 4 390 females ( 61.23% ), which had a median age of 56 (interquartile range, 10) years. The median frailty score was 2 (interquartile range, 3 ) among the study subjects, and the median frailty score was 2 ( interquartile range, 2 ) among residents at ages of 45 to 59 years, and 2 (interquartile range, 3) among residents at ages of 60 to 69 years. The overall detection of frailty was 16.07%, and the detection of frailty was 13.52% among subjects at ages of 45 to 59 years and 21.01% among subjects at ages of 60 to 69 years. Multivariable logistic regression analysis identified physical activity ( OR=0.826, 95%CI: 0.719-0.949 ) and sleep quality ( OR: 3.376-11.493, 95%CI: 2.907-15.808 ) as factors affecting frailty among middle-aged and elderly residents. Following age stratification, physical activity ( OR=0.817, 95%CI: 0.681-0.981 ) and sleep quality ( OR: 3.076-11.566, 95%CI: 2.518-18.216 ) as factors affecting frailty among subjects at ages of 45 to 59 years, while sleep quality ( OR: 3.777-11.827, 95%CI: 3.002-18.547 ) significantly correlated with frailty among residents at ages of 60 to 69 years. Conclusion Physical activity and sleep quality are associated with the risk of frailty among middle-aged and elderly populations.

Objective:To investigate the changes in cognitive and behavioral function in patients with cerebral small vessel disease and their correlation with serum homocysteine (Hcy) and cystatin C (CysC) levels.Methods:Ninety-four patients with cerebral small vascular disease who received treatment in Zhejiang Xin'an International Hospital from March 2019 to March 2021 were included in the observation group. An additional 80 healthy controls who concurrently received physical examination were included in the control group. The Mini-Mental State Scale (MMSE) and the Montreal Assessment Scale (MoCA) were used to evaluate cognitive function. The activity of daily living (ADL) was used to evaluate behavioral function. Enzyme-linked immunosorbent assay was used to measure serum Hcy and CysC levels. MMSE score, MoCA score, ADL score, serum Hcy and CysC levels were compared between the two groups. The receiver operating characteristic (ROC) curve was used to analyze the sensitivity and specificity of serum Hcy and CysC levels in the diagnosis of cerebral small vessel disease. Pearson correlation analysis was performed to analyze the correlation between serum Hcy and CysC levels and cognitive behavioral function.Results:MMSE score, MoCA score, and ADL score in the observation group were (21.21 ± 2.87) points, (20.18 ± 3.25) points, and (34.32 ± 5.45) points, which were significantly lower than (28.23 ± 1.02) points, (28.45 ± 0.89) points and (76.35 ± 8.29) points in the control group ( t = 20.78, 22.05, 40.03, all P < 0.05). Serum Hcy and CysC in the observation group were (16.28 ± 2.97) μmol/L and (1.21 ± 0.23) mg/L, which were significantly higher than (8.35 ± 1.76) μmol/L and (0.56 ± 0.12) mg/L in the control group ( t = 20.95, 22.77, both P < 0.05). ROC curve analysis showed that the sensitivity and specificity of serum Hcy in diagnosing cerebral small vessel disease were 85.29% and 53.85%, respectively; the sensitivity and specificity of serum CysC in diagnosing cerebral small vessel disease were 81.54% and 41.38%, respectively. Pearson correlation analysis revealed that serum Hcy level was linearly negatively correlated with MMSE score, MoCA score, and ADL score ( r = -0.79, -0.68, -0.71, all P < 0.05); serum CysC level was linearly negatively correlated with MMSE score, MoCA score, and ADL score ( r = -0.67, -0.75, -0.78, P < 0.05). Conclusion:The cognitive and behavioral function of patients with cerebral small vessel disease are decreased, and serum Hcy and CysC levels are increased. Serum Hcy and CysC levels are closely related to the cognitive and behavioral function of patients with cerebral small vessel disease.

Objective:To explore the associations between perceived built environment attributes and adults’ leisure-time physical activity in four cities of China.Methods:Multistage cluster random sampling method was used to select adults aged 25 to 64 in Hangzhou, Suzhou, Chengdu, and Qingdao. Data were collected from June 2017 to July 2018. The perception of the urban built environment was assessed by the neighborhood environment walkability scale-abbreviated (NEWS-A), and the physical activity was assessed by the International Physical Activity Questionnaire. Generalized linear mixed models were used to explore the relationship between the perceived built environment and leisure-time physical activities.Results:A total of 3 789 participants were included in the analysis. After adjusting for potential confounders, better access to public services ( OR=1.34, 95% CI: 1.02-1.75) and higher aesthetic quality ( OR=1.37, 95% CI: 1.09-1.73) were positively associated with the possibility of engaging in leisure-time physical activity in the past week. Similarly, these two attributes were positively associated with leisure-time walking. Higher scores on the perception of street connectivity were positively associated with leisure-time walking [ exp( β)=1.09, 95% CI: 1.00-1.19]. Higher residential density [ exp( β)=1.000 4, 95% CI:1.000 0-1.000 8], better access to physical activity destinations[ exp( β)=1.09, 95% CI: 1.00-1.19], and better aesthetics [ exp( β)=1.11, 95% CI:1.00-1.22] were associated with higher leisure-time physical activity. Similarly, these three attributes were positively associated with the possibility of meeting the WHO recommendations. Conclusion:Changing some urban built environment attributes may increase leisure-time physical activity.

CD8+T cells are critical immune cells protecting the body against infection and cancer. Long-lived memory CD8+T cells formed in a prior infection can reproduce to mount a faster and stronger im-mune response at a second encounter with the cognate antigen. The activation, clonal expansion and re-sponse of T cells are energetically demanding processes tightly coupled in cellular metabolism. Meanwhile, changes in cellular metabolism could also affect the development of memory T cells following acute infection. In this review, we discussed the current understanding of the mechanism by which glycometabolic pathways manipulate the differentiation of memory CD8+T cells in order to provide reference for improving vaccine de-velopment and cancer treatment.

Influenza vaccination is the most effective means to prevent and control influenza epi-demics, and the universal influenza vaccine which can induce broad and long-term protective effect is still in its infancy. Up to date, several broad neutralizing antibodies that can antagonize a variety of influenza A vi-ruses have been identified, and the crystal structure of the antibody and HA complex reveals at least three highly conserved epitopes. Deep insights into the molecular mechanism of the interactions between neutrali-zing antibodies and virus antigen can elicit a new strategy not only for rational design of universal influenza vaccines, but also for the development of antibody-based therapeutics against influenza virus. In this paper, advances in research of universal neutralizing antibodies and vaccines for Influenza A virus are reviewed.