Objective To investigate the correlation mechanism between miR-153-3p and Nrf2 expression in human nucleus pulposus cells and intervertebral disc degeneration(IDD).Methods The oxidative damage model of nucleus pulposus cells was duplicated induced by H2O2.MiR-153-3p inhibitor-NC,si-NRF2-NC,miR-153-3p inhibitor,and si-NRF2 were transfected into nucleus pulposus cells according to the grouping require.The transfection efficiency was detected by RT-qPCR and Western blot.The cell viability was determined by the CCK-8 assay,when the intracellular reactive oxygen species(ROS)levels and the ratio of mitochondrial membrane potential decline were measured by flow cytometry,RT-qPCR was used to detect the expression levels of Nrf2,MMP-3,Col II,PINK1,Parkin,P62,and p38 MAPK.And dual luciferase reporter assay was used to measure luciferase activity.Results(1)HNPCs treated with H2O2 showed a decrease in HNPCs cell viability,a reduction in mitochondrial membrane potential,an increase in ROS levels,and a decrease in Col II expression(P<0.05).And the expression of MMP-3,P62,and p38 MAPK increased,while the expression of PINK1 and Parkin decreased.There was no significant change in Nrf2(P>0.05).(2)Inhibition of miR-153-3p expression in nucleus pulposus cells treated with H2O2 led to increased cell viability,elevated mitochondrial membrane potential,reduced ROS levels,and enhanced Col-II expression,accompanied by decreased expression of MMP-3,P62,and p38 MAPK,while simultaneously increasing the expression of PINK1,Parkin,and Nrf2(P<0.05).(3)When miR-153-3p expression was inhibited and Nrf2 expression was silenced in nucleus pulposus cells treated with H2O2,a notable decline in cell viability and mitochondrial membrane potential was observed,along with a marked increase in ROS levels.Additionally,Col-II expression decreased,whereas the expression of MMP-3,P62,and p38 MAPK increased.However,the expression of Nrf2,PINK1,and Parkin decreased(P<0.05).(4)Dual-luciferase assay analysis revealed binding sites and a binding relationship between miR-153-3p and Nrf2,indicating a negative correlation between miR-153-3p and Nrf2(P<0.05).Conclusion Inhibition of miR-153-3p expression can alleviate H2O2 induced degeneration and fibrosis of nucleus pulposus cells,activate autophagy of damaged nucleus pulposus cells,and delay apoptosis of nucleus pulposus cells.This effect is related to the PINK1/Parkin pathway and p38 MAPK inflammatory response pathway regulated by Nrf2.

AIM:To investigate the effects of high-dose methotrexate(MTX)on alkaline phosphatase(ALP)and the effects of ALP changes on bone me-tabolism,bone marrow granulogram function,liver function and excretion.METHODS:Aspartate ami-notransferase(AST),alanine aminotransferase(ALT)and albumin(ALB)were used as liver function indi-cators,serum calcium(Ca)and phosphorus(P)were used as bone metabolism indicators,neutro-phil(ANC)and white blood cell count(WBC)were used as bone marrow granuloline function indica-tors,and methotrexate C48h concentration ≥1 μmol/L was used as the excretion delay.One-way ANOVA analysis was performed on the ALP levels before and after the first chemotherapy and the second chemotherapy,and the children were divided into normal group and low group according to the ALP level,and the seven indexes before and after che-motherapy were quantitatively and qualitatively an-alyzed,and univariate and multivariate Logistic re-gression analysis was performed on the concentra-tion of methotrexate C48h and the above indexes in the children treated with the second chemothera-py.RESULTS:After the first chemotherapy and the second chemotherapy,ALP was significantly de-creased[(204.0±83.6)U/L vs.(172.8±67.3)U/L,(179.4±59.3)U/L vs.(169.6±57.1)U/L,all P＜0.05],and the serum Ca,P,ANC,WBC,and ALB were sig-nificantly decreased(P＜0.05),and AST and ALT were increased(P＜0.05),and ALT was an indepen-dent risk factor for delayed excretion(OR=1.049,95％CI 1.023-1.077,P＜0.001),ALB was an indepen-dent protective factor for delayed excretion(OR=0.551,95％CI 0.460-0.660,P＜0.001),and ALP was not a significant contributor to MTX excretion de-lay.CONCLUSION:ALP is not a good predictor of liv-er function and bone marrow granulopathy func-tion due to a significant decrease in ALP caused by high-dose MTX,and ALP together with serum calci-um and phosphorus levels can constitute an early warning indicator of bone metabolism disorders.

Purpose To compare the 5.0T gradient echo coronary magnetic resonance angiography(CMRA)using compressed sensing(CS)acceleration technology(5.0TCS-CMRA)vs.3.0T gradient echo-CMRA(3.0TCS-CMRA)using CS.Materials and Methods Twenty-five healthy volunteers aged 23 to 30 years from December 16,2023 to January 14,2024 at Peking Union Medical College Hospital,Chinese Academy of Medical Sciences were prospective enrolled in this study.The interval between 3.0TCS-CMRA and 5.0TCS-CMRA was within two weeks.3.0TCS-CMRA used T2 preparation and 5.0TCS-CMRA did not use T2 preparation.The image quality scores,coronary artery length,signal-to-noise ratio(SNR)and contrast-to-noise ratio between coronary blood and adjacent myocardium or tissue(CNRmyo-blood)were evaluated.Results On 5.0TCS-CMRA,the SNR and CNRmyo-blood of the proximal right coronary artery(RCA)in 25 healthy volunteers were significantly higher than those of 3.0TCS-CMRA(SNR:318.07±94.06 vs.223.81±51.19,t=-5.609,P<0.001;CNRmyo-blood:212.75±91.44 vs.149.70±59.53,Z=-3.619,P<0.001),while the SNR and CNRmyo-blood of proximal left anterior descending coronary artery(LAD)and left circumflex coronary artery(LCX)were not significantly higher than those of 3.0TCS-CMRA(SNR:315.52±102.49 vs.306.35±92.85,t=-0.627,P=0.536;289.72±88.79 vs.272.87±84.68,t=-1.226,P=0.232;CNRmyo-blood:135.83±93.53 vs.203.94±74.30,t=4.132,P<0.001;117.66±79.63 vs.161.60±78.91,t=3.127,P=0.005).The length of the three coronary arteries measured by 5.0TCS-CMRA was significantly shorter than that of 3.0TCS-CMRA[RCA:(126.04±31.54)mm vs.(137.20±29.93)mm,t=2.911,P=0.008;LAD:(122.68±24.63)mm vs.(134.24±23.38)mm,Z=-3.026,P=0.002;LCX:(57.07±26.70)mm vs.(68.27±24.02)mm,t=2.552,P=0.018].There was no significant difference in the scanning time required between 3.0TCS-CMRA and 5.0TCS-CMRA[(8.60±2.84)min vs.(8.30±2.32)min,Z=-0.183,P=0.855].The image scores of the three major coronary arteries of 5.0TCS-CMRA were significantly lower than those of 3.0TCS-CMRA(RCA:2.52±0.59 vs.3.16±0.69,Z=-3.258,P=0.001;LAD:2.72±0.74 vs.3.24±0.66,Z=-2.540,P=0.011;LCX:2.44±0.71 vs.3.00±0.87,Z=-2.462,P=0.014).Conclusion In the absence of T2 preparation,5.0TCS-CMRA can still show obvious advantages in the SNR and CNRmyo-blood of proximal RCA compared with 3.0TCS-CMRA,which suggests the application potential of 5.0TCS-CMRA.In the future,a suitable T2 preparation pulse or potential alternative may significantly improve the performance of the 5.0TCS-CMRA.

Acute pancreatitis(AP),one of the most common acute abdominal conditions in clinical practice,is typically self-limiting.However,approximately 20%of cases progress to severe acute pancreatitis,characterized by persistent systemic inflammatory response syndrome and multiple-organ dysfunction syndrome,with a high mortality rates.The pathogenesis of AP involves complex pathophysiological processes,and in recent years,the role of oxidative stress(OS)in AP has garnered increasing attention.OS refers to an imbalance between reactive oxygen species production and antioxidant capacity following endogenous or exogenous stimuli,which can lead to pancreatic cell injury,exacerbation of inflammatory responses,and organ dysfunction.Notably,antioxidants have demonstrated efficacy in reducing OS-induced pancreatic damage and multi-organ dysfunction in animal models.This article reviews current molecular mechanisms of OS in AP,its role in disease progression and recent advances in antioxidant-based therapeutic strategies for AP.

Objective:To investigate the regulatory effect of zinc finger transcription factor 580 (ZNF580) gene on autophagy and extracellular matrix (ECM) secretion in human pancreatic cancer cells PANC1.Methods:PANC1 cells were transfected with 500 ng/ml short hairpin RNA-ZNF580 (shRNA-ZNF580) and a ZNF580 expression vector with a green fluorescent protein reporter gene (GFP-ZNF580) using lentiviral transfection to establish the ZNF580-silenced group and ZNF580-overexpression group, respectively. PANC1 cells were treated with 10 mmol/L rapamycin (RA), a cell autophagy inducer, and the autophagy inhibitor LY294002 for 2 hours to construct the autophagy-induced group and autophagy-inhibited group, respectively. The autophagy inhibition+ZNF580 silencing group was established by transfecting PANC1 cells with 500 ng/ml sh-ZNF580 using lentiviral transfection while simultaneously adding 10 mmol/L LY294002. PANC1 cells cultured in conventional medium served as control group. The expression levels of ZNF580 protein and autophagy-related proteins ATG7 and LC3 in PANC1 cells from each group were detected by Western blot. The expression changes of ECM secretion-related markers type I collagen (Col-Ⅰ), Col-Ⅲ, fibronectin (FN), and tumor necrosis factor-α (TNF-α) in PANC1 cells were measured by ELISA.Results:Compared with control group, the protein expression levels of ATG7, LC3-Ⅰ, and LC3-Ⅱ in PANC1 cells of the ZNF580-silenced group were significantly decreased (0.40±0.04 vs 0.81±0.13, 0.66±0.08 vs 2.0±0.45, 0.78±0.10 vs 1.89±0.23), while they were significantly increased in the ZNF580-overexpression group (2.07±0.17 vs 0.83±0.09, 1.21±0.37 vs 0.88±0.09, 0.77±0.16 vs 0.37±0.06). The protein expression level of ZNF580 in PANC1 cells of the autophagy inhibition group was significantly down-regulated compared with the control group (0.40±0.15 vs 1.07±0.18), while it was significantly up-regulated in the autophagy induction group (1.59±0.25 vs 0.67±0.09). Compared with the control group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells were significantly decreased in the ZNF580-silenced group (5.02±0.81 vs 8.38±0.83, 6.17±0.83 vs 10.73±1.69, 28.66±2.47 vs 45.20±4.31, 10.09±1.32 vs 19.48±2.77), which were significantly increased in the ZNF580-overexpression group (19.28±2.05 vs 8.38±0.83, 28.29±5.96 vs 10.73±1.69, 103.22±6.37 vs 45.20±4.31, 46.78±6.96 vs 19.48±2.77), significantly decreased in the autophagy inhibition group (5.10±0.66 vs 9.01±1.24, 7.22±0.67 vs 11.83±1.71, 28.45±2.82 vs 43.51±4.38, 12.16±2.13 vs 20.53±3.65, respectively), and significantly increased in the autophagy induction group (20.49±3.68 vs 9.01±1.24, 26.58±3.96 vs 11.83±1.71, 73.18±7.15 vs 43.51±4.38, 41.11±8.87 vs 20.53±3.65). Compared with the autophagy inhibition group and the ZNF580-silenced group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells of the autophagy inhibition+ZNF580 silencing group were significantly decreased (Col-Ⅰ: 3.36±1.25 vs 5.73±0.62 and 5.57±0.35; Col-Ⅲ: 4.15±0.16 vs 6.24±0.90 and 6.71±0.34; FN: 18.31±2.00 vs 26.46±1.18 and 27.09±2.01; TNF-α: 6.81±0.46 vs 9.96±1.87 and 10.62±0.65). All the above differences were statistically significant (all P value <0.05). Conclusions:The transcription factor ZNF580 could positively regulate the levels of autophagy and ECM secretion in PANC1 cells. The combined application of ZNF580 gene silencing and autophagy inhibitors can significantly inhibit ECM secretion in PANC1 cells.

AIM:To investigate the effects of high-dose methotrexate(MTX)on alkaline phosphatase(ALP)and the effects of ALP changes on bone me-tabolism,bone marrow granulogram function,liver function and excretion.METHODS:Aspartate ami-notransferase(AST),alanine aminotransferase(ALT)and albumin(ALB)were used as liver function indi-cators,serum calcium(Ca)and phosphorus(P)were used as bone metabolism indicators,neutro-phil(ANC)and white blood cell count(WBC)were used as bone marrow granuloline function indica-tors,and methotrexate C48h concentration ≥1 μmol/L was used as the excretion delay.One-way ANOVA analysis was performed on the ALP levels before and after the first chemotherapy and the second chemotherapy,and the children were divided into normal group and low group according to the ALP level,and the seven indexes before and after che-motherapy were quantitatively and qualitatively an-alyzed,and univariate and multivariate Logistic re-gression analysis was performed on the concentra-tion of methotrexate C48h and the above indexes in the children treated with the second chemothera-py.RESULTS:After the first chemotherapy and the second chemotherapy,ALP was significantly de-creased[(204.0±83.6)U/L vs.(172.8±67.3)U/L,(179.4±59.3)U/L vs.(169.6±57.1)U/L,all P＜0.05],and the serum Ca,P,ANC,WBC,and ALB were sig-nificantly decreased(P＜0.05),and AST and ALT were increased(P＜0.05),and ALT was an indepen-dent risk factor for delayed excretion(OR=1.049,95％CI 1.023-1.077,P＜0.001),ALB was an indepen-dent protective factor for delayed excretion(OR=0.551,95％CI 0.460-0.660,P＜0.001),and ALP was not a significant contributor to MTX excretion de-lay.CONCLUSION:ALP is not a good predictor of liv-er function and bone marrow granulopathy func-tion due to a significant decrease in ALP caused by high-dose MTX,and ALP together with serum calci-um and phosphorus levels can constitute an early warning indicator of bone metabolism disorders.

Acute pancreatitis(AP),one of the most common acute abdominal conditions in clinical practice,is typically self-limiting.However,approximately 20%of cases progress to severe acute pancreatitis,characterized by persistent systemic inflammatory response syndrome and multiple-organ dysfunction syndrome,with a high mortality rates.The pathogenesis of AP involves complex pathophysiological processes,and in recent years,the role of oxidative stress(OS)in AP has garnered increasing attention.OS refers to an imbalance between reactive oxygen species production and antioxidant capacity following endogenous or exogenous stimuli,which can lead to pancreatic cell injury,exacerbation of inflammatory responses,and organ dysfunction.Notably,antioxidants have demonstrated efficacy in reducing OS-induced pancreatic damage and multi-organ dysfunction in animal models.This article reviews current molecular mechanisms of OS in AP,its role in disease progression and recent advances in antioxidant-based therapeutic strategies for AP.

Purpose To compare the 5.0T gradient echo coronary magnetic resonance angiography(CMRA)using compressed sensing(CS)acceleration technology(5.0TCS-CMRA)vs.3.0T gradient echo-CMRA(3.0TCS-CMRA)using CS.Materials and Methods Twenty-five healthy volunteers aged 23 to 30 years from December 16,2023 to January 14,2024 at Peking Union Medical College Hospital,Chinese Academy of Medical Sciences were prospective enrolled in this study.The interval between 3.0TCS-CMRA and 5.0TCS-CMRA was within two weeks.3.0TCS-CMRA used T2 preparation and 5.0TCS-CMRA did not use T2 preparation.The image quality scores,coronary artery length,signal-to-noise ratio(SNR)and contrast-to-noise ratio between coronary blood and adjacent myocardium or tissue(CNRmyo-blood)were evaluated.Results On 5.0TCS-CMRA,the SNR and CNRmyo-blood of the proximal right coronary artery(RCA)in 25 healthy volunteers were significantly higher than those of 3.0TCS-CMRA(SNR:318.07±94.06 vs.223.81±51.19,t=-5.609,P<0.001;CNRmyo-blood:212.75±91.44 vs.149.70±59.53,Z=-3.619,P<0.001),while the SNR and CNRmyo-blood of proximal left anterior descending coronary artery(LAD)and left circumflex coronary artery(LCX)were not significantly higher than those of 3.0TCS-CMRA(SNR:315.52±102.49 vs.306.35±92.85,t=-0.627,P=0.536;289.72±88.79 vs.272.87±84.68,t=-1.226,P=0.232;CNRmyo-blood:135.83±93.53 vs.203.94±74.30,t=4.132,P<0.001;117.66±79.63 vs.161.60±78.91,t=3.127,P=0.005).The length of the three coronary arteries measured by 5.0TCS-CMRA was significantly shorter than that of 3.0TCS-CMRA[RCA:(126.04±31.54)mm vs.(137.20±29.93)mm,t=2.911,P=0.008;LAD:(122.68±24.63)mm vs.(134.24±23.38)mm,Z=-3.026,P=0.002;LCX:(57.07±26.70)mm vs.(68.27±24.02)mm,t=2.552,P=0.018].There was no significant difference in the scanning time required between 3.0TCS-CMRA and 5.0TCS-CMRA[(8.60±2.84)min vs.(8.30±2.32)min,Z=-0.183,P=0.855].The image scores of the three major coronary arteries of 5.0TCS-CMRA were significantly lower than those of 3.0TCS-CMRA(RCA:2.52±0.59 vs.3.16±0.69,Z=-3.258,P=0.001;LAD:2.72±0.74 vs.3.24±0.66,Z=-2.540,P=0.011;LCX:2.44±0.71 vs.3.00±0.87,Z=-2.462,P=0.014).Conclusion In the absence of T2 preparation,5.0TCS-CMRA can still show obvious advantages in the SNR and CNRmyo-blood of proximal RCA compared with 3.0TCS-CMRA,which suggests the application potential of 5.0TCS-CMRA.In the future,a suitable T2 preparation pulse or potential alternative may significantly improve the performance of the 5.0TCS-CMRA.

Objective:To investigate the regulatory effect of zinc finger transcription factor 580 (ZNF580) gene on autophagy and extracellular matrix (ECM) secretion in human pancreatic cancer cells PANC1.Methods:PANC1 cells were transfected with 500 ng/ml short hairpin RNA-ZNF580 (shRNA-ZNF580) and a ZNF580 expression vector with a green fluorescent protein reporter gene (GFP-ZNF580) using lentiviral transfection to establish the ZNF580-silenced group and ZNF580-overexpression group, respectively. PANC1 cells were treated with 10 mmol/L rapamycin (RA), a cell autophagy inducer, and the autophagy inhibitor LY294002 for 2 hours to construct the autophagy-induced group and autophagy-inhibited group, respectively. The autophagy inhibition+ZNF580 silencing group was established by transfecting PANC1 cells with 500 ng/ml sh-ZNF580 using lentiviral transfection while simultaneously adding 10 mmol/L LY294002. PANC1 cells cultured in conventional medium served as control group. The expression levels of ZNF580 protein and autophagy-related proteins ATG7 and LC3 in PANC1 cells from each group were detected by Western blot. The expression changes of ECM secretion-related markers type I collagen (Col-Ⅰ), Col-Ⅲ, fibronectin (FN), and tumor necrosis factor-α (TNF-α) in PANC1 cells were measured by ELISA.Results:Compared with control group, the protein expression levels of ATG7, LC3-Ⅰ, and LC3-Ⅱ in PANC1 cells of the ZNF580-silenced group were significantly decreased (0.40±0.04 vs 0.81±0.13, 0.66±0.08 vs 2.0±0.45, 0.78±0.10 vs 1.89±0.23), while they were significantly increased in the ZNF580-overexpression group (2.07±0.17 vs 0.83±0.09, 1.21±0.37 vs 0.88±0.09, 0.77±0.16 vs 0.37±0.06). The protein expression level of ZNF580 in PANC1 cells of the autophagy inhibition group was significantly down-regulated compared with the control group (0.40±0.15 vs 1.07±0.18), while it was significantly up-regulated in the autophagy induction group (1.59±0.25 vs 0.67±0.09). Compared with the control group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells were significantly decreased in the ZNF580-silenced group (5.02±0.81 vs 8.38±0.83, 6.17±0.83 vs 10.73±1.69, 28.66±2.47 vs 45.20±4.31, 10.09±1.32 vs 19.48±2.77), which were significantly increased in the ZNF580-overexpression group (19.28±2.05 vs 8.38±0.83, 28.29±5.96 vs 10.73±1.69, 103.22±6.37 vs 45.20±4.31, 46.78±6.96 vs 19.48±2.77), significantly decreased in the autophagy inhibition group (5.10±0.66 vs 9.01±1.24, 7.22±0.67 vs 11.83±1.71, 28.45±2.82 vs 43.51±4.38, 12.16±2.13 vs 20.53±3.65, respectively), and significantly increased in the autophagy induction group (20.49±3.68 vs 9.01±1.24, 26.58±3.96 vs 11.83±1.71, 73.18±7.15 vs 43.51±4.38, 41.11±8.87 vs 20.53±3.65). Compared with the autophagy inhibition group and the ZNF580-silenced group, the levels of extracellularly secreted Col-Ⅰ, Col-Ⅲ, FN, and TNF-α in PANC1 cells of the autophagy inhibition+ZNF580 silencing group were significantly decreased (Col-Ⅰ: 3.36±1.25 vs 5.73±0.62 and 5.57±0.35; Col-Ⅲ: 4.15±0.16 vs 6.24±0.90 and 6.71±0.34; FN: 18.31±2.00 vs 26.46±1.18 and 27.09±2.01; TNF-α: 6.81±0.46 vs 9.96±1.87 and 10.62±0.65). All the above differences were statistically significant (all P value <0.05). Conclusions:The transcription factor ZNF580 could positively regulate the levels of autophagy and ECM secretion in PANC1 cells. The combined application of ZNF580 gene silencing and autophagy inhibitors can significantly inhibit ECM secretion in PANC1 cells.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.