ObjectiveTo explore the host immunomodulatory effect and its molecular mechanism of gallic acid （GA） in Bacillus Calmette-Guérin （BCG） infected macrophages， and to provide experimental evidence for host-directed anti-tuberculosis research against Mycobacterium tuberculosis （Mtb）. MethodsThe minimum inhibitory concentration （MIC） of GA against BCG was determined by resazurin colorimetric assay. Bacterial growth-curve test and MTT colorimetric assay were used to evaluate the effects of GA on BCG proliferation and invasion. A BCG-infected RAW264.7 macrophage model was established， including infection control group （Ctrl）， isoniazid （INH） group， and low-， medium- and high-dose GA groups. Flow cytometry combined with colony-plate counting was performed to detect intracellular BCG survival. Cell apoptosis was measured by Annexin V-APC/7-AAD double staining， and qRT-PCR was used to detect the mRNA expression of apoptosis-related genes Caspase-3 and Bcl-2. Macrophage polarization phenotypes were analyzed by flow cytometry. Intracellular reactive oxygen species （ROS） were detected using DHE fluorescent probe. The Griess assay was applied to determine supernatant nitric oxide （NO） concentration. ELISA was used to detect the levels of TNF-α， IFN-γ， IL-6 and IL-10 in cell culture supernatant. ResultsThe MIC of GA against BCG was 1 250 μmol/L. GA inhibited BCG proliferation and invasion in a concentration-dependent manner （P＜0.05）. Safe concentrations of 20， 40 and 80 μmol/L were selected for subsequent cellular experiments. Thirty-six-hour GA pretreatment markedly reduced the intracellular BCG bacterial load （P＜0.05）. GA exerted bidirectional regulation on macrophage apoptosis： it inhibited apoptosis at the early-stage infection and promoted apoptosis at the late-stage infection. GA induced M1-type polarization in early infection and facilitated M2-type polarization in late infection （P＜0.05）. Meanwhile， GA decreased intracellular ROS and NO levels， down-regulated the secretion of pro-inflammatory cytokines and up-regulated the secretion of anti-inflammatory cytokine IL-10 （P＜0.05）. ConclusionGA inhibits BCG proliferation and invasion in vitro. It remodels host immune microenvironment via bidirectional regulation of macrophage apoptosis and polarization as well as anti-oxidative and anti-inflammatory effects， showing potential value for host-directed anti-mycobacterial therapy.

ObjectiveTo explore the host immunomodulatory effect and its molecular mechanism of gallic acid （GA） in Bacillus Calmette-Guérin （BCG） infected macrophages， and to provide experimental evidence for host-directed anti-tuberculosis research against Mycobacterium tuberculosis （Mtb）. MethodsThe minimum inhibitory concentration （MIC） of GA against BCG was determined by resazurin colorimetric assay. Bacterial growth-curve test and MTT colorimetric assay were used to evaluate the effects of GA on BCG proliferation and invasion. A BCG-infected RAW264.7 macrophage model was established， including infection control group （Ctrl）， isoniazid （INH） group， and low-， medium- and high-dose GA groups. Flow cytometry combined with colony-plate counting was performed to detect intracellular BCG survival. Cell apoptosis was measured by Annexin V-APC/7-AAD double staining， and qRT-PCR was used to detect the mRNA expression of apoptosis-related genes Caspase-3 and Bcl-2. Macrophage polarization phenotypes were analyzed by flow cytometry. Intracellular reactive oxygen species （ROS） were detected using DHE fluorescent probe. The Griess assay was applied to determine supernatant nitric oxide （NO） concentration. ELISA was used to detect the levels of TNF-α， IFN-γ， IL-6 and IL-10 in cell culture supernatant. ResultsThe MIC of GA against BCG was 1 250 μmol/L. GA inhibited BCG proliferation and invasion in a concentration-dependent manner （P＜0.05）. Safe concentrations of 20， 40 and 80 μmol/L were selected for subsequent cellular experiments. Thirty-six-hour GA pretreatment markedly reduced the intracellular BCG bacterial load （P＜0.05）. GA exerted bidirectional regulation on macrophage apoptosis： it inhibited apoptosis at the early-stage infection and promoted apoptosis at the late-stage infection. GA induced M1-type polarization in early infection and facilitated M2-type polarization in late infection （P＜0.05）. Meanwhile， GA decreased intracellular ROS and NO levels， down-regulated the secretion of pro-inflammatory cytokines and up-regulated the secretion of anti-inflammatory cytokine IL-10 （P＜0.05）. ConclusionGA inhibits BCG proliferation and invasion in vitro. It remodels host immune microenvironment via bidirectional regulation of macrophage apoptosis and polarization as well as anti-oxidative and anti-inflammatory effects， showing potential value for host-directed anti-mycobacterial therapy.

Objective To investigate the correlation between hemoglobin A1c(HbA1c)levels and excessive daytime sleepiness(EDS)in patients with type 2 diabetes mellitus(T2DM).Methods A total of 132 T2DM patients and 40 healthy people(NC group)who were treated in the outpatient department of Nanjing Meishan Hospital from December 2020 to December 2022 were selected.General clinical data of the subjects were collected,and their Epworth sleepiness scale(ESS)scores,Pittsburgh sleep quality index(PSQI),and apnea-hypopnea index(AHI)were measured.Based on ESS scores,T2DM patients were divided into simple T2DM group(ESS score<9,n=99)and EDS group(ESS score≥9,n=33)according to ESS score.The baseline data were compared for each group.Spearman correlation analysis and multivariate logistic regression model were used to evaluate the correlation between HbA1c levels and EDS.The receiver operating characteristic(ROC)curve was used to calculate the area under the curve(AUC)to evaluate the predictive value of HbA1c levels for EDS.Results HbA1c,fasting plasma glucose,AHI index and PSQI score in EDS group were higher than those in T2DM and NC group(P<0.05).Spearman correlation analysis showed that ESS score was positively correlated with HbA1c in T2DM patients(P<0.05).Multivariate logistic regression analysis revealed that elevated HbA1c levels emerged as a significant and independent risk factor for the onset of EDS.The ROC curve indicated that the AUC of HbA1c for predicting EDS was 0.736.Conclusions There is an independent positive correlation between HbA1c levels and EDS in T2DM patients,which provides clues for early identification and treatment of EDS in clinical practice.

Objective:To investigate the correlation between serum neuron-specific enolase (NSE) levels and poor neurological outcomes in cardiac arrest (CA) patients supported by veno-arterial extracorporeal membrane oxygenation (VA-ECMO).Methods:This retrospective analysis was conducted on adult CA patients treated with VA-ECMO at Hangzhou First People's Hospital Affiliated to Westlake University School of Medicine, and Second Affiliated Hospital Zhejiang University School of Medicine, from December 2018 to February 2024. General clinical data and serial serum NSE levels at 24, 48, and 72 h after ECMO initiation were collected. Based on the Glasgow-Pittsburgh Cerebral Performance Category (CPC) at discharge, patients were divided into poor neurological outcome group (CPC 3-5) and good neurological outcome group (CPC 1-2). Differences in serum NSE levels between the two groups were compared. The accuracy of serum NSE levels at three time points in predicting poor neurological outcomes in CA patients was assessed via receiver operating characteristic curves, and the optimal cut-off values were determined by the Youden index. Multivariate logistic regression analysis was performed to determine the relationship between serum NSE levels and poor neurological outcomes. Subgroup analysis was based on age, sex, location of CA, and extracorporeal cardiopulmonary resuscitation (ECPR).Results:A total of 120 eligible CA patients were included, with 88 patients (73.3%) having poor neurological outcomes at discharge. Serum NSE levels at 24, 48, and 72 h after ECMO initiation were higher in the poor outcome group compared to the good outcome group (all P<0.05). The serum NSE level at 72 h had the highest accuracy in predicting poor outcomes, with an area under the curve (AUC) of 0.91 (95% CI: 0.85-0.96), and a cut-off value of 42.0 μg/L. The AUCs for 24 and 48 h were 0.78 (95% CI: 0.69-0.86) and 0.87 (95% CI: 0.80-0.94), with cut-off values of 70.6 μg/L and 64.5 μg/L, respectively. Multivariate logistic regression analysis suggested that the serum NSE level at 72 h was associated with poor outcomes ( P<0.05), and an NSE level >42.0 μg/L was an independent risk factor for poor outcomes ( OR=20.29, 95% CI: 2.90-92.15). Subgroup analysis showed that serum NSE level at 72 h was an independent risk factor for poor neurological outcomes in CA patients aged<60 years old, male or female, out-of-hospital or in-hospital CA, and whether to perform ECPR (all P<0.05). Conclusion:Elevated serum NSE levels at 72 h after VA-ECMO initiation are associated with poor neurological outcomes in CA patients, with the cut-off value of 42.0 μg/L.

Objective To investigate the correlation between hemoglobin A1c(HbA1c)levels and excessive daytime sleepiness(EDS)in patients with type 2 diabetes mellitus(T2DM).Methods A total of 132 T2DM patients and 40 healthy people(NC group)who were treated in the outpatient department of Nanjing Meishan Hospital from December 2020 to December 2022 were selected.General clinical data of the subjects were collected,and their Epworth sleepiness scale(ESS)scores,Pittsburgh sleep quality index(PSQI),and apnea-hypopnea index(AHI)were measured.Based on ESS scores,T2DM patients were divided into simple T2DM group(ESS score<9,n=99)and EDS group(ESS score≥9,n=33)according to ESS score.The baseline data were compared for each group.Spearman correlation analysis and multivariate logistic regression model were used to evaluate the correlation between HbA1c levels and EDS.The receiver operating characteristic(ROC)curve was used to calculate the area under the curve(AUC)to evaluate the predictive value of HbA1c levels for EDS.Results HbA1c,fasting plasma glucose,AHI index and PSQI score in EDS group were higher than those in T2DM and NC group(P<0.05).Spearman correlation analysis showed that ESS score was positively correlated with HbA1c in T2DM patients(P<0.05).Multivariate logistic regression analysis revealed that elevated HbA1c levels emerged as a significant and independent risk factor for the onset of EDS.The ROC curve indicated that the AUC of HbA1c for predicting EDS was 0.736.Conclusions There is an independent positive correlation between HbA1c levels and EDS in T2DM patients,which provides clues for early identification and treatment of EDS in clinical practice.

Objective To investigate the impact of hemoglobin(Hb)level on the risk of all-cause mortality in elderly patients with stable coronary artery disease(SCAD)treated by percutaneous coronary intervention(PCI).Methods A retrospective cohort study was conducted on 195 elderly SCAD patients treated by PCI in our hospital from June 2016 to December 2017,a total of 180 case were followed up.They were divided into four groups based on quartiles of Hb levels:Q1 group[≤122(108.28±12.53)g/L,n=47],Q2 group[122＜Hb≤137(130.07±4.33)g/L,n=43],Q3 group[137＜Hb≤148(142.67±3.10)g/L,n=46],and Q4 group[＞148(158.36±8.50)g/L,n=44].Their clinical data were collected and compared among the groups.All patients were followed up,with all-cause mortality as endpoint.Cox regression analysis was used to evaluate the impact of Hb level on all-cause mortality in elderly SCAD patients treated by PCI.Results For the 180 patients with complete follow-up,the median follow-up time was 703(415,1121)d,and the incidence of all-cause mortality was 10.00％(totally 18 deaths).And the incidence was 25.50％(12 deaths),7.00％(3 death),4.30％(2 deaths)and 2.30％(1 death)in Q1,Q2,Q3,and Q4 groups,respectively,with statistically significant differences(P＜0.01).Univariate Cox regression analysis showed age was a risk factor for all-cause death(P＜0.01),while Hb level,BMI,smoking history,dyslipidemia,and aspirin use were a risk facotr for all-cause death(P＜0.05,P＜0.01).Further multivariate Cox regression analysis revealed Hb level was a risk facotr for all-cause death(HR=0.96,95％CI:0.93-0.99,P＜0.01).The risk of all-cause mortality was 3.68 times higher in the Q1 group than the Q2 group(HR=3.68,95％CI:1.01-13.45,P＜0.05),and there was no correlation in the incidence between the Q3 and Q4 groups(P＞0.05).Conclusion Low Hb level is an independent predictor for all-cause mortality in elderly SCAD pa-tients after PCI.

Objective:To investigate the prognostic significance of anemia in elderly patients with stable coronary artery disease treated by percutaneous coronary intervention(PCI).Methods:In this retrospective cohort study, 180 patients with stable coronary artery disease aged ≥60 years undergone percutaneous coronary intervention and with complete clinical data, admitted to Nanjing Meishan Hospital between June 2016 and December 2017, were selected.Baseline clinical data of the patients were collected, including hemoglobin, C-reactive protein, lipids, fasting glucose, glycated hemoglobin, blood creatinine, and left ventricular ejection fraction on cardiac color ultrasound.The endpoints of the follow-up included major adverse cardiac events(MACE), such as all-cause death, non-fatal myocardial infarction, and non-fatal stroke.According to the hemoglobin level, participants were divided into an anemia group(n=32)and a non-anemia group(n=148). Clinical data of the two groups were compared, and the Kaplan-Meier method and multivariate analysis with the Cox regression method were used to evaluate the effect of anemia on the occurrence of MACE in elderly patients with stable coronary artery disease treated by PCI.Results:The median duration of the follow-up of the 180 patients was 702.5 days and MACE occurred in 27(15.0%). Compared with the non-anemia group, the anemia group had a higher age, C-reactive protein level and MACE ratio, lower levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol and the glomerular filtration rate, and higher proportions of patients with smoking history and patients with dyslipidemia( P<0.05). Kaplan-Meier curves suggested that the incidence of MACE in the anemia group was higher than that in the non-anemia group[37.5%(12/32) vs.10.1%(15/148), P<0.05]. Multivariate analysis with the Cox regression method showed that the risk of MACE in the anemia group was 2.91 times higher than that in the non-anemia group( HR=2.91, 95% CI: 1.13-7.48, P<0.05). Conclusions:Anemia is an independent predictor of MACE in elderly patients with stable coronary artery disease after PCI.

Objective:To summarize and analyze the clinical and genetic characteristics of Chinese patients with adrenomyeloneuropathy (AMN).Methods:Clinical data were collected and analyzed retrospectively on AMN patients who were diagnosed by genetic testing in Shanghai Sixth People′s Hospital, Shanghai Jiao Tong University School of Medicine from May 2008 to August 2022. Clinical characteristics of AMN patients with different types of gene mutations were compared. Loe score was used to evaluate the severity of white matter demyelinating, and the serum levels of very long-chain fatty acids (VLCFA) in patients with or without white matter demyelinating were compared. The motor function of the AMN patients was assessed using the Expanded Disability Status Scale (EDSS), and the association between EDSS scores and the course of disease was analyzed.Results:A total of 23 male patients with onset age of (29.52±9.91) years were included in this study. The first symptom of all patients was abnormal lower extremities, of which 17 patients showed stiffness and weakness in their lower limbs (73.9%, 17/23), and 6 patients showed numbness and pain in both lower limbs (26.1%, 6/23). The occurrence of symptoms was not related to the type of gene mutation. White matter demyelination occurred in 33.3% (7/21) of patients over a disease duration of (7.67±4.46) years. There was no statistically significant difference in serum VLCFA level between the white-matter demyelination group and the non-demyelination group. The EDSS score was positively correlated with the disease duration ( r=0.57, P=0.006). Sixteen ABCD1 gene mutations were found in this study, among which c.5_19delinsTCTCCAGG (p.P2Lfs *12) was reported for the first time. Four probands belonging to different families carried the c.1415_1416del (p.Q472Rfs *83) variant. Conclusions:Lower limb movement disorders and sensory dysfunction are the prominent clinical manifestations in AMN patients, with deterioration of motor function associated with the course of disease. AMN may be converted to cerebral type and VLCFA concentration is not associated with the phenotypic changes. The c.1415_1416del (p.Q472Rfs *83) mutation is a hot spot mutation of the disease.

Laughing gas (Nitrogen monoxide) is currently abused due to its low price and easy availability. This article discussed the clinical manifestations of a patient with subacute combined degeneration of the spinal cord caused by inhalation of laughing gas. The patient developed numbness of extremities, unstable walking, and decreased serum vitamin B 12 level. MRI of the cervical spine showed abnormal signals in the lateral and posterior cords of the cervical spinal cord (C2-6) , neuroelectrophysiological examination showed peripheral nerve damage in the extremities. After treatment with vitamin B 12 supplementation, the patient's condition gradually improved. Clinicians diagnose subacute combined degeneration of the spinal cord, especially when the patient has no gastrointestinal disease, diet, malnutrition, etc., they need to carefully inquire about the history of nitrous oxide inhalation to avoid missed diagnosis.

Laughing gas (Nitrogen monoxide) is currently abused due to its low price and easy availability. This article discussed the clinical manifestations of a patient with subacute combined degeneration of the spinal cord caused by inhalation of laughing gas. The patient developed numbness of extremities, unstable walking, and decreased serum vitamin B 12 level. MRI of the cervical spine showed abnormal signals in the lateral and posterior cords of the cervical spinal cord (C2-6) , neuroelectrophysiological examination showed peripheral nerve damage in the extremities. After treatment with vitamin B 12 supplementation, the patient's condition gradually improved. Clinicians diagnose subacute combined degeneration of the spinal cord, especially when the patient has no gastrointestinal disease, diet, malnutrition, etc., they need to carefully inquire about the history of nitrous oxide inhalation to avoid missed diagnosis.