AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.

Objective:To investigate the association between blood selenium levels and sex hormones in Chinese men aged 18-79 years.Methods:Data were derived from the China National Human Biomonitoring survey conducted in 2017-2018, with a final sample size of 5 414 men. General demographic characteristics, behavioral habits, and dietary frequency were collected through questionnaires and physical examinations. Fasting blood samples were collected to measure blood lead, serum testosterone, and estradiol levels. Complex sampling linear regression models were used to analyze the associations between blood selenium levels and testosterone, estradiol, and the testosterone/estradiol ratio, adjusting for confounding factors including age, education level, marital status, smoking status, alcohol consumption, seafood intake, soy product intake, protein supplement intake, BMI, and diabetes status.Results:The mean age of the 5 414 participants was (46.85±27.91) years; 4 774 (91.65%) were of Han ethnicity and 4 505 (86.68%) were married. The median ( Q1, Q3) blood selenium concentration in men was 97.80 (80.64, 116.99) μg/L. After adjusting for confounding factors, the complex sampling linear regression model revealed negative associations between blood selenium levels and both testosterone levels and the testosterone/estradiol ratio, with a significant linear trend ( Ptrend<0.05). Compared with the Q1 group, the β (95% CI) values for testosterone in the Q2, Q3, and Q4 groups were -0.02 (-0.06 to 0.02), -0.03 (-0.08 to 0.01), and -0.06 (-0.09 to -0.02), respectively. Similarly, the β (95% CI) values for the testosterone/estradiol ratio in the Q2, Q3, and Q4 groups were -0.01 (-0.03 to 0.02), -0.01 (-0.04 to 0.04), and -0.03 (-0.06 to -0.01), respectively. Subgroup analysis indicated stronger associations between blood selenium levels and testosterone/estradiol levels in non-smoking and obese men (BMI≥28 kg/m2). Conclusion:Blood selenium levels are negatively associated with testosterone levels and the testosterone/estradiol ratio in Chinese adult males.

AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.

Objective:To investigate the association between blood selenium levels and sex hormones in Chinese men aged 18-79 years.Methods:Data were derived from the China National Human Biomonitoring survey conducted in 2017-2018, with a final sample size of 5 414 men. General demographic characteristics, behavioral habits, and dietary frequency were collected through questionnaires and physical examinations. Fasting blood samples were collected to measure blood lead, serum testosterone, and estradiol levels. Complex sampling linear regression models were used to analyze the associations between blood selenium levels and testosterone, estradiol, and the testosterone/estradiol ratio, adjusting for confounding factors including age, education level, marital status, smoking status, alcohol consumption, seafood intake, soy product intake, protein supplement intake, BMI, and diabetes status.Results:The mean age of the 5 414 participants was (46.85±27.91) years; 4 774 (91.65%) were of Han ethnicity and 4 505 (86.68%) were married. The median ( Q1, Q3) blood selenium concentration in men was 97.80 (80.64, 116.99) μg/L. After adjusting for confounding factors, the complex sampling linear regression model revealed negative associations between blood selenium levels and both testosterone levels and the testosterone/estradiol ratio, with a significant linear trend ( Ptrend<0.05). Compared with the Q1 group, the β (95% CI) values for testosterone in the Q2, Q3, and Q4 groups were -0.02 (-0.06 to 0.02), -0.03 (-0.08 to 0.01), and -0.06 (-0.09 to -0.02), respectively. Similarly, the β (95% CI) values for the testosterone/estradiol ratio in the Q2, Q3, and Q4 groups were -0.01 (-0.03 to 0.02), -0.01 (-0.04 to 0.04), and -0.03 (-0.06 to -0.01), respectively. Subgroup analysis indicated stronger associations between blood selenium levels and testosterone/estradiol levels in non-smoking and obese men (BMI≥28 kg/m2). Conclusion:Blood selenium levels are negatively associated with testosterone levels and the testosterone/estradiol ratio in Chinese adult males.

Objective:To establish the prediction model for postoperative delirium in elderly patients with mild stroke undergoing non-cardiac and non-intracranial surgery.Methods:This was a nested case-control study. Seven hundred and fifty elderly patients of either sex with mild stroke, aged ≥65 yr, undergoing elective surgical procedures under general anesthesia in the Department of Gastrointestinal Surgery, Orthopedics and Urology at the Tangshan Workers Hospital from May to December 2023, were selected. The perioperative clinical data were collected. The incidence of postoperative delirium was assessed using the Confusion Assessment Scale 1-7 days after surgery or 1 day before discharge. The patients were assigned to the training set and the validation set in a ratio of 7∶3 using a simple random sampling method. Multivariate logistic regression was used to identify the risk factors for postoperative delirium, a postoperative delirium risk prediction model was established based on the risk factors, the nomogram was developed, and the receiver operating characteristic (ROC) curve, calibration curve and decision curve were plotted to assess the accuracy of the prediction model. The prediction model was verified using the validation set, and the calibration curve and ROC curve were plotted to assess the predictive performance of the model.Results:A total of 721 patients were finally included, and 108 patients developed postoperative delirium. Older age, high American Society of Anesthesiologists Physical Status classification, history of preoperative hypertension, short years of education, high preoperative Pittsburgh sleep quality index score, high preoperative National Institutes of Health Stroke Scale score, high intraoperative hypothermia, intraoperative hypotension and high postoperative numerical rating scale score were independent risk factors for postoperative delirium ( P<0.05). The area under the ROC curve of the training set prediction model was 0.996, with a sensitivity of 1.000, and specificity of 0.945. The slope of the calibration curve was close to 1, and the predicted risk of postoperative delirium was in good agreement with the actual risk. When the threshold probability of the decision curve was 0-0.9, the net return rate was higher than the null line. Validation set: In the calibration curve of the prediction model, the cohort and calibration curves were close to the ideal line, with an area under the ROC curve of 0.997, sensitivity of 1.000, and specificity of 0.962. Conclusions:Based on age, American Society of Anesthesiologists Physical Status classification, history of preoperative hypertension, years of education, preoperative Pittsburgh sleep quality index score, National Institutes of Health Stroke Scale score, intraoperative hypothermia and hypotension and postoperative numerical rating scale score, the prediction model for postoperative delirium is developed and has a good predictive performance in elderly patients with mild stroke undergoing non-cardiac and non-intracranial surgery.

Objective To study the effects of different extracorporeal circulation temperature combined with dexmedetomidine on oxidative stress in patients undergoing cardiac surgery under cardiopulmonary bypass.Methods 240 patients who underwent cardiac surgery under cardiopulmonary bypass were selected from June 2021 to June 2024.The patients were 50 to 75 years old and did not have severe hepatic or renal insufficiency.The Mini-Mental State Examination was completed 1 day before the operation.Patients were routinely established for cardiopulmonary bypass.Patients were randomly divided into four groups(n=55):hypothermia+normal saline group(L0),hypothermia+dexmedetomidine group(L1),hyperthermia+normal saline group(H0)and hyper-thermia+dexmedetomidine group(H1).The nasopharyngeal temperature was maintained at(30±1)℃in the hypothermia group and(33±1)℃in the high-temperature group during the reflux period.Dexmedetomidine injection was intravenously injected at 1 μg/kg 10 minutes before anesthesia in L1 and H1 groups,and pumped continuously at a rate of 0.5 μg/(kg·h)until the end of surgery.L0 and H0 groups were given equal volume of normal saline until the end of operation.5 mL of central venous blood was collected before the beginning of anesthesia(T1),at the end of surgery(T2),24 h(T3)and 48 h(T4)after surgery,and serum neuron specific enolase(NSE),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum were determined by xanthine oxidase method and thiobarbituric acid method.The Confusion Assessment Method-ICU was used to evaluate the occurrence of POD.Results 220 patients were finally enrolled,including 81 patients in POD group,with an incidence of 36.8%.Compared with NPOD group,the concentrations of NSE,IL-6,TNF-ɑ and MDA in POD group were in-creased,while the concentration of SOD was decreased.Compared with L0 group,the concentrations of NSE,IL-6,TNF-ɑ and MDA were decreased and the concentration of SOD increased in L1 group.Compared with H0 group,NSE,IL-6,TNF-ɑ and MDA concentrations in H1 group were decreased,while SOD concentration was in-creased.Compared with L0 group,concentration of NSE,IL-6,TNF-ɑ and MDA increased,while the concentra-tion of SOD decreased in H0 group.Compared with L1 group,concentration of NSE,IL-6,TNF-ɑ and MDA in-creased,while the concentration of SOD decreased in H1 group.Conclusion Hypothermia combined with dexme-detomidine during cardiopulmonary bypass can reduce oxidative stress response and POD in patients.

Objective To study the effects of different extracorporeal circulation temperature combined with dexmedetomidine on oxidative stress in patients undergoing cardiac surgery under cardiopulmonary bypass.Methods 240 patients who underwent cardiac surgery under cardiopulmonary bypass were selected from June 2021 to June 2024.The patients were 50 to 75 years old and did not have severe hepatic or renal insufficiency.The Mini-Mental State Examination was completed 1 day before the operation.Patients were routinely established for cardiopulmonary bypass.Patients were randomly divided into four groups(n=55):hypothermia+normal saline group(L0),hypothermia+dexmedetomidine group(L1),hyperthermia+normal saline group(H0)and hyper-thermia+dexmedetomidine group(H1).The nasopharyngeal temperature was maintained at(30±1)℃in the hypothermia group and(33±1)℃in the high-temperature group during the reflux period.Dexmedetomidine injection was intravenously injected at 1 μg/kg 10 minutes before anesthesia in L1 and H1 groups,and pumped continuously at a rate of 0.5 μg/(kg·h)until the end of surgery.L0 and H0 groups were given equal volume of normal saline until the end of operation.5 mL of central venous blood was collected before the beginning of anesthesia(T1),at the end of surgery(T2),24 h(T3)and 48 h(T4)after surgery,and serum neuron specific enolase(NSE),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels were detected by ELISA.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in serum were determined by xanthine oxidase method and thiobarbituric acid method.The Confusion Assessment Method-ICU was used to evaluate the occurrence of POD.Results 220 patients were finally enrolled,including 81 patients in POD group,with an incidence of 36.8%.Compared with NPOD group,the concentrations of NSE,IL-6,TNF-ɑ and MDA in POD group were in-creased,while the concentration of SOD was decreased.Compared with L0 group,the concentrations of NSE,IL-6,TNF-ɑ and MDA were decreased and the concentration of SOD increased in L1 group.Compared with H0 group,NSE,IL-6,TNF-ɑ and MDA concentrations in H1 group were decreased,while SOD concentration was in-creased.Compared with L0 group,concentration of NSE,IL-6,TNF-ɑ and MDA increased,while the concentra-tion of SOD decreased in H0 group.Compared with L1 group,concentration of NSE,IL-6,TNF-ɑ and MDA in-creased,while the concentration of SOD decreased in H1 group.Conclusion Hypothermia combined with dexme-detomidine during cardiopulmonary bypass can reduce oxidative stress response and POD in patients.

Objective:To investigate the effects of urinary nickel exposure on insulin resistance, islet function and diabetes risk in adults aged 18 years and above in China.Methods:Based on the China National Human Biomonitoring from 2017 to 2018, a total of 500 diabetes patients were randomly selected as the case group, and the matched euglycemic control were selected by 1∶1 matching ratio. The urinary and venous blood samples of the subjects were collected, and the urinary nickel levels and serum glucose metabolism indexes such as fasting blood glucose and fasting insulin were detected, and the insulin resistance index (HOMA-IR), β cell function index (HOMA-β), and adjusted HOMA-β were calculated by using homeostasis model assessment. A multivariate logistic regression model was used to analyze the association between urinary nickel level and diabetes risk. Multiple linear regression models were used to evaluate the association of urinary nickel level with HOMA-IR, HOMA-β and adjusted HOMA-β.Results:The sex ratio of controls and cases was 1∶1. The multivariate logistic regression model showed that after adjusting for factors such as education level, smoking status, alcohol consumption, rice and meat intakes, family history of diabetes, BMI, total cholesterol level, hypertension, and urinary creatinine, compared with T1 group, the ORs of diabetes risk in the T2 and T3 groups were 1.36 (95% CI: 0.98-1.89) and 1.60 (95% CI: 1.14-2.24), respectively. The multiple linear regression model showed a positive association between urinary nickel levels and the elevated HOMA-IR, the β value of HOMA-IR in the T3 group was 0.12 (95% CI: 0.01-0.25) compared with the T1 group and each one-unit increase in the log-transformed urinary nickel level (2.71 μg/L) was associated with a 0.06 elevation in HOMA-IR (95% CI: 0.02-0.10). Meanwhile, the urinary nickel levels were negative associated with the adjusted HOMA-β, the β value of adjusted HOMA-β in the T3 group were -0.26 compared with the T1 group (95% CI: -0.41- -0.11), and each one-unit increase in the log-transformed urinary nickel level (2.71 μg/L) was associated with a -0.09 decrease in adjusted HOMA-β(95% CI: -0.14- -0.04). Conclusion:Higher urinary nickel level is positively correlated with elevated insulin resistance and diabetes risk while negatively correlated with the function of pancreatic islet β cells in adults in China.

Approximately 15% of couples suffer from infertility worldwide, and male factors contribute to about 30% of total sterility cases. However, there is little progress in treatments due to the obscured understanding of underlying mechanisms. Recently microRNAs have emerged as a key player in the process of spermatogenesis. Expression profiling of miR-181a was carried out in murine testes and spermatocyte culture system. In vitro cellular and biochemical assays were used to examine the effect of miR-181a and identify its target S6K1, as well as elucidate the function with chemical inhibitor of S6K1. Human testis samples analysis was employed to validate the findings. miR-181a level was upregulated during mouse spermatogenesis and knockdown of miR-181a attenuated the cell proliferation and G1/S arrest and increased the level of S6K1, which was identified as a downstream target of miR-181a. Overexpression of S6K1 also led to growth arrest of spermatocytes while inhibitor of S6K1 rescued the miR-181a knockdown-mediated cell proliferation defect. In human testis samples of azoospermia patients, low level of miR-181a was correlated with defects in the spermatogenic process. miR-181a is identified as a new regulator and high level of miR-181a contributes to spermatogenesis via targeting S6K1.

Approximately 15% of couples suffer from infertility worldwide, and male factors contribute to about 30% of total sterility cases. However, there is little progress in treatments due to the obscured understanding of underlying mechanisms. Recently microRNAs have emerged as a key player in the process of spermatogenesis. Expression profiling of miR-181a was carried out in murine testes and spermatocyte culture system. In vitro cellular and biochemical assays were used to examine the effect of miR-181a and identify its target S6K1, as well as elucidate the function with chemical inhibitor of S6K1. Human testis samples analysis was employed to validate the findings. miR-181a level was upregulated during mouse spermatogenesis and knockdown of miR-181a attenuated the cell proliferation and G1/S arrest and increased the level of S6K1, which was identified as a downstream target of miR-181a. Overexpression of S6K1 also led to growth arrest of spermatocytes while inhibitor of S6K1 rescued the miR-181a knockdown-mediated cell proliferation defect. In human testis samples of azoospermia patients, low level of miR-181a was correlated with defects in the spermatogenic process. miR-181a is identified as a new regulator and high level of miR-181a contributes to spermatogenesis via targeting S6K1.