AIM:Regulatory T cells(Tregs)are a specialized subset of CD4+T cells primarily involved in im-munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen-tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un-explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1fl/fl)mice and Treg-specific AMPKα1 knockout mice(AMPKα1fl/flFoxp3cre mice).RESULTS:Compared to AMPKα1fl/fl mice,the percentage of eosinophils in the bone marrow of AMPKα1fl/flFoxp3cre mice was significant-ly reduced.Additionally,while the thymus of AMPKα1fl/flFoxp3cre mice exhibited normal structure,both its size and the ra-tio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduc-tion in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4+T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1fl/flFoxp3cre mice compared to AMPKα1fl/fl mice.Importantly,the proportion of Siglec-F+CD11b+eosinophils in the thymus was significantly lower in AMPKα1fl/flFoxp3cre mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4+T cells in peripheral blood,alongside a decrease in the proportion of mature CD8+T cells.Similar-ly,the proportion of CD4+T cells in the spleen of AMPKα1fl/flFoxp3cre mice was elevated compared to AMPKα1fl/fl mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1fl/flFoxp3cre mice.In lymph nodes,the medullary boundaries in AMPKα1fl/flFoxp3cre mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1fl/fl mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3+T cell population,particularly the CD8+T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1fl/flFoxp3cre mice,the percentage of CD4+T cells was markedly in-creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1fl/flFoxp3cre and AMPKα1fl/fl mice.CONCLUSION:The populations of mature Tregs,CD8+T cells and eosinophils in various im-mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.

Objective This study aimed to evaluate the interaction between antiretroviral drug efavirenz and anti-tuberculosis 1H3P3(isoniazid plus rifapentine)in people living with HIV.Methods HIV-positive individuals on efavirenz-containing(600 mg)antiretroviral therapy(ART)received 1H3P3 regimen containing rifapentine(450 mg)plus isoniazid(400 mg)3 times a week for 1 month.Efavirenz concentrations were measured at weeks 0,2,4,8.Rifapentine concentration was determined at weeks 2 and 4.HIV RNA load was determined at weeks 0 and 8.Treatment target was efavirenz concentration＞1 mg/L.The anti-TB prevention was considered acceptable if the target of efavirenz concentration was achieved in more than 80％of participants.The participants were followed up for 18 months to evaluate the efficacy of treatment.Results Thirty-one participants living with HIV were enrolled in the study.Two participants were excluded from PK analysis because his/her baseline efavirenz concentration＜1 mg/L,suggesting poor treatment adherence.Evaluable PK data were available for 29 participants,including 23(79.3％)males.The median[interquartile range(IQR)]age of the participants was 43.0(32.5,53.5)years.The median(IQR)efavirenz plasma concentration was 2.33(1.96,2.34)mg/L at week 0,2.32(1.90,3.28)mg/L at week 2,2.07(1.83,3.09)mg/L at week 4,and 2.71(2.14,3.33)mg/L at week 8.Efavirenz concentration did not show significant difference between the 4 time points(P＞0.05).Median(IQR)rifapentine concentration was 9.36(6.23,16.47)mg/L at week 2,and 9.36(6.41,15.56)mg/L at week 4.Rifapentine concentration did not show significant difference between week 2 and week 4(P＞0.05).Efavirenz concentrations was＞1 mg/L in all participants at weeks 2,4,and 8.Furthermore,efavirenz concentration was significantly higher in females and patients with body weight＜60 kg compared with males and those with body weight ≥60 kg(P＜0.05).None of the participants had symptoms or signs of active tuberculosis during 18-month follow-up.Conclusions Isoniazid plus rifapentine(1H3P3 regimen)did not have significant effect on the plasma concentrations of efavirenz.

Objective This study aimed to evaluate the interaction between antiretroviral drug efavirenz and anti-tuberculosis 1H3P3(isoniazid plus rifapentine)in people living with HIV.Methods HIV-positive individuals on efavirenz-containing(600 mg)antiretroviral therapy(ART)received 1H3P3 regimen containing rifapentine(450 mg)plus isoniazid(400 mg)3 times a week for 1 month.Efavirenz concentrations were measured at weeks 0,2,4,8.Rifapentine concentration was determined at weeks 2 and 4.HIV RNA load was determined at weeks 0 and 8.Treatment target was efavirenz concentration＞1 mg/L.The anti-TB prevention was considered acceptable if the target of efavirenz concentration was achieved in more than 80％of participants.The participants were followed up for 18 months to evaluate the efficacy of treatment.Results Thirty-one participants living with HIV were enrolled in the study.Two participants were excluded from PK analysis because his/her baseline efavirenz concentration＜1 mg/L,suggesting poor treatment adherence.Evaluable PK data were available for 29 participants,including 23(79.3％)males.The median[interquartile range(IQR)]age of the participants was 43.0(32.5,53.5)years.The median(IQR)efavirenz plasma concentration was 2.33(1.96,2.34)mg/L at week 0,2.32(1.90,3.28)mg/L at week 2,2.07(1.83,3.09)mg/L at week 4,and 2.71(2.14,3.33)mg/L at week 8.Efavirenz concentration did not show significant difference between the 4 time points(P＞0.05).Median(IQR)rifapentine concentration was 9.36(6.23,16.47)mg/L at week 2,and 9.36(6.41,15.56)mg/L at week 4.Rifapentine concentration did not show significant difference between week 2 and week 4(P＞0.05).Efavirenz concentrations was＞1 mg/L in all participants at weeks 2,4,and 8.Furthermore,efavirenz concentration was significantly higher in females and patients with body weight＜60 kg compared with males and those with body weight ≥60 kg(P＜0.05).None of the participants had symptoms or signs of active tuberculosis during 18-month follow-up.Conclusions Isoniazid plus rifapentine(1H3P3 regimen)did not have significant effect on the plasma concentrations of efavirenz.

AIM:Regulatory T cells(Tregs)are a specialized subset of CD4+T cells primarily involved in im-munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen-tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un-explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1fl/fl)mice and Treg-specific AMPKα1 knockout mice(AMPKα1fl/flFoxp3cre mice).RESULTS:Compared to AMPKα1fl/fl mice,the percentage of eosinophils in the bone marrow of AMPKα1fl/flFoxp3cre mice was significant-ly reduced.Additionally,while the thymus of AMPKα1fl/flFoxp3cre mice exhibited normal structure,both its size and the ra-tio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduc-tion in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4+T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1fl/flFoxp3cre mice compared to AMPKα1fl/fl mice.Importantly,the proportion of Siglec-F+CD11b+eosinophils in the thymus was significantly lower in AMPKα1fl/flFoxp3cre mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4+T cells in peripheral blood,alongside a decrease in the proportion of mature CD8+T cells.Similar-ly,the proportion of CD4+T cells in the spleen of AMPKα1fl/flFoxp3cre mice was elevated compared to AMPKα1fl/fl mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1fl/flFoxp3cre mice.In lymph nodes,the medullary boundaries in AMPKα1fl/flFoxp3cre mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1fl/fl mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3+T cell population,particularly the CD8+T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1fl/flFoxp3cre mice,the percentage of CD4+T cells was markedly in-creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1fl/flFoxp3cre and AMPKα1fl/fl mice.CONCLUSION:The populations of mature Tregs,CD8+T cells and eosinophils in various im-mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.

Objective:To observe the application effect of a self-designed assisted abdominal breathing rehabilitation device in elderly patients undergoing postoperative chemotherapy for lung cancer.Methods:Self-designed an assisted abdominal breathing rehabilitation device. Used the convenience sampling method, 116 elderly patients admitted to the First Affiliated Hospital of Anhui Medical University from April 2021 to April 2022 undergoing postoperative chemotherapy for lung cancer were selected as study subjects, and they were divided into control and observation groups by the random number table method, with 58 cases in each group. The control group received routine abdominal breathing training, and the observation group used auxiliary abdominal breathing rehabilitation device for abdominal breathing training, and compared the training compliance rate, lung function indexes and six minutes-walk test scores of the two groups before and after 2 weeks of training.Results:After 2 weeks of training, the training compliance rate of patients in the observation group was 100.00% (58/58), which was higher than 87.93% (51/58) in the control group, and the difference was statistically significant ( χ 2=7.45, P<0.05). After 2 weeks of training, FEV 1, FVC, FEV 1/FVC were (1.81 ± 0.29) L, (1.98 ± 0.32) L, (91.91 ± 5.98) % respectively in the observation group, and (1.49 ± 0.31) L, (1.73 ± 0.41) L, (87.11 ± 9.44) % respectively in the control group, there were statistically significant differences between the two groups ( t=-4.13, -2.60, -2.36, all P<0.05). After 2 weeks of training, the six minutes-walk test score was (0.86 ± 0.71) points in the observation group and (1.02 ± 0.74) points in the control group, and the difference was statistically significant ( t=2.05, P<0.05). Conclusions:An assisted abdominal breathing rehabilitation device helps to improve the training compliance rate, enhance lung function recovery and improve exercise endurance in elderly patients undergoing chemotherapy after lung cancer surgery.

Objective:To explore the expression and clinical significance of immunosuppressive receptor T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) on the peripheral blood mononuclear cells (PBMC) in silicosis patients with Mycobacterium tuberculosis infection. Methods:August 2018, a total of 78 patients with silicosis (all were quarry workers in Sanmen County, Zhejiang Province) were enrolled and divided into silicosis combined with active pulmonary tuberculosis group (APTB group), silicosis combined with latent tuberculosis infection group (LTBI group), and simple silicosis with non-tuberculosis infection group (non-TB group). Flow cytometry was used to analyze the expressions of TIGIT, programmed death-1 (PD-1) and transcription factor T-bet on PBMC from patients. Mann-Whitney U test and Pearson correlations analysis were used for statistical analysis. Results:Among the 78 patients, eight were in the APTB group, 24 in the LTBI group, and 46 in the non-TB group. The expressions of PD-1 and TIGIT on CD8 + T cells in the APTB group (29.45%(16.78%) and 65.40%(12.12%), respectively) were significantly higher than those in the LTBI group (17.40%(11.17%) and 48.30%(28.75%), respectively; U=23.500 and 43.500, respectively, P=0.000 8 and 0.020 5, respectively) and non-TB group (15.95%(12.46%) and 45.30%(19.75%), respectively; U=64.000 and 69.000, respectively, P=0.002 3 and 0.003 8, respectively), and the differences were all statistically significant. The expression of TIGIT was positively correlated with PD-1 on CD8 + T cells in silicosis patients ( r=0.434 3, P<0.01). The proportion of PD-1 + TIGIT + CD8 + T cells in the APTB group (19.90%(22.67%)) was significantly higher than those in the non-TB group (11.55%(11.29%), U=76.500, P=0.007 1) and LTBI group (11.55%(10.53%), U=41.000, P=0.015 4), while the proportion of PD-1 -TIGIT -CD8 + T cells in the APTB group (30.60%(12.90%)) was significantly lower than non-TB group (48.90%(18.98%), U=58.000, P=0.001 3) and LTBI group (47.20%(24.59%), U=41.000, P=0.015 4). The differences were all statistically significant. The expression of T-bet on the peripheral blood CD8 + T cells in the APTB group (29.45%(16.78%)) was higher than that in the non-TB group (15.95%(12.46%)) and the LTBI group (17.40%(11.17%)), and the differences were both statistically significant ( U=46.500 and 46.000, respectively, P=0.000 3 and 0.028 3, respectively). The expression of T-bet on CD8 + T cells was positively correlated with TIGIT on CD8 + T cells ( r=0.456 7, P<0.01). The expression of T-bet on PD-1 + TIGIT + CD8 + T cells in the APTB group (65.40%(12.12%)) was higher than those in the LTBI group (48.30%(28.75%), U=23.500, P=0.000 8) and non-TB group (45.30%(19.75%), U=65.000, P=0.002 6), and the differences were both statistically significant. Conclusion:The immunosuppressive receptor PD-1 and TIGIT are highly expressed on CD8 + T cells in silicosis patients with active pulmonary tuberculosis, which indicates CD8 + T cells exhaustion in these population, while the highly co-expression of T-bet suggests the exhausted subsets may have reversed potentiality.

Objective:To explore the clinical effect of nebulized acetylcysteine inhalation combined with bronchoscopy in treating elderly patients with severe ventilator-associated pneumonia (VAP).Methods:From January 2016 to December 2017, 80 elderly patients with severe VAP who were hospitalized in Zhejiang Hospital were divided into acetylcysteine+ bronchoscopy group [24 males and 16 females, aged (78±7) years] and bronchoscopy group [26 males and 14 females, aged (80±7) years]using random number table for a prospective cohort study. Patients in bronchoscopy group were treated with bronchoscopy in addition to conventional supportive care of symptoms. Patients in acetylcysteine+ bronchoscopy group received nebulized acetylcysteine inhalation therapy on the basis of the treatment given to patients in bronchoscopy group. Patients in both groups received treatment continuously for 7 days. The simplified clinical pulmonary infection score (CPIS) in both groups was assessed before and after treatment. Venous blood of 10 mL was collected before and after treatment to detect leukocyte count, serum C-reactive protein, and procalcitonin. Arterial blood of 1 mL was collected before and after treatment to detect partial arterial oxygen pressure (PaO 2), partial arterial carbon dioxide pressure (PaCO 2), oxygenation index. The inhalation platform pressure (Pplat), dynamic lung compliance (Cdyn), airway resistance, work of breathing, time of antibiotic use, and time of mechanical ventilation of patients in two groups were recorded before and after treatment. Data were statistically analyzed with chi-square test and t test. Results:(1) The simplified CPIS, leukocyte count, serum C-reactive protein, and procalcitonin of patients in acetylcysteine+ bronchoscopy group were significantly lower than those in bronchoscopy group after treatment ( t=2.32, 2.15, 6.08, 7.12, P<0.05 or P<0.01). The simplified CPIS, leukocyte count, serum C-reactive protein, and procalcitonin of patients in acetylcysteine+ bronchoscopy group and bronchoscopy group after treatment were significantly lower than those before treatment ( t=13.76, 13.60, 12.70, 8.32, 11.44, 14.28, 9.48, 9.50, P<0.01). (2) Compared with bronchoscopy group, patients in acetylcysteine+ bronchoscopy group had significantly higher PaO 2 and oxygenation index ( t=4.14, 2.55, P<0.05 or P<0.01) but significantly lower PaCO 2 ( t=4.36, P<0.01) after treatment. The PaO 2 and oxygenation index of patients in acetylcysteine+ bronchoscopy group after treatment were significantly higher than those before treatment ( t=10.90, 43.72, P<0.01). The PaO 2 and oxygenation index of patients in bronchoscopy group after treatment were also significantly higher than those before treatment ( t=6.55, 43.03, P<0.01). The PaCO 2 of patients in both groups after treatment were significantly lower than those before treatment ( t=21.54, 21.92, P<0.01). (3) The Cdyn of patients in acetylcysteine+ bronchoscopy group after treatment was significantly higher than that in bronchoscopy group ( t=5.41, P<0.01), and Pplat, airway resistance, and work of breathing were significantly lower than those in bronchoscopy group ( t=2.18, 5.46, 2.49, P<0.05 or P<0.01). The Cdyn of patients in both groups after treatment were significantly higher than those before treatment ( t=16.10, 10.90, P<0.01), and Pplat, airway resistance, and work of breathing were significantly lower than those before treatment ( t=21.18, 11.13, 9.32, 15.50, 5.17, 5.97, P<0.01). (4)The time of mechanical ventilation and antibiotic usage of patients in acetylcysteine+ bronchoscopy group were (6.9±1.9)and (8.7±2.8) d, respectively, which were significantly shorter than (10.1±2.2) and (11.6±3.5) d in bronchoscopy group ( t=6.85, 4.09, P<0.01). Conclusions:Nebulized acetylcysteine inhalation combined with bronchoscopy can significantly control the degree of lung infection in elderly patients with severe VAP, improve patients′ respiratory mechanics parameter and blood gas analysis indicator, and shorten the time for mechanical ventilation and antibiotic usage.

Objective To retrospectively analyze the clinical and laboratory characteristics of patients with positive blood culture results for Mycobacterium tuberculosis (M.tb).Methods The clinical laboratory database of patients suspected with disseminated tuberculosis from January 2009 to January 2017 in Huashan Hospital affiliated with Fudan University were collected and analyzed.The clinical manifestations,laboratory characteristics and outcomes between disseminated tuberculosis patients with positive blood culture (positive blood culture group) for M.tb and negative results (negative blood culture group) were compared.T test,Mann-Whitney U test and Fisher exact test were used for statistical analysis.Results A total of 5 589 patients suspected with M.tb infection had peripheral blood culture for mycobacterium.Positive blood culture for M.tb was found in 26 disseminated tuberculosis patients,while 6 patients finally identified as nontuberculous mycobacterium (NTM) with species identification,and 22 disseminated tuberculosis patients with negative blood culture results were enrolled during the same period as control.The mean age ([49.1 ± 10.1] years old vs [38.3 ± 17.1] years old,t =2.460,P =0.018),the proportion of diagnosed with fever of unknown origin at admission (FUO) (65.0％ [13/20] vs 13.6％ [3/22],P =0.001),the proportion of diagnosed with focal infection (30.0％ [6/20] vs 86.4％ [19/22],P =0.001),the proportion of patients with other diseases (75.0％[15/20] vs 22.7％ [5/22],P =0.002),the proportion of patients with hematological diseases (35.0％ [7/20] vs 4.5％ [1/22],P =0.018) and the proportion of patients with tumor (20％ [4/20] vs 0[0/22],P =0.043) in the positive blood culture group were significantly different from those in the negative blood culture group.Laboratory examinations of the percentage of neutrophils,the percentage of lymphocytes,the percentage of monocytes,the value of neutrophil/lymphocyte,the level of hemoglobin,the level of erythrocyte sedimentation rate,the level of C-reactive protein,the level of procalcitonin and the positive rate of T-SPOT.TB in positive blood culture groups were significantly different from those in negative blood culture group (all P ＜ 0.05).Conclusions Peripheral blood M.tb culture is more likely to be positive for those elder disseminated tuberculosis patients with hematological diseases or tumors,and those with increase of neutrophil counts and inflammation markers but reduction of lymphocyte counts and hemoglobin.

Objective To investigate the prevalence of tuberculosis among silicosis patients and silica exposure patients,and to analysis the risk factors of tuberculosis among these population.Methods A total of 1 227 silica exposure patients from Wenling,Zhejiang were enrolled in this field study.Basic demographic information was collected and chest X-ray was taken for each patient.Sputum was collected for Mycobacterium tuberculosis culture and strain identification. In univariate analysis,t test was performed for continuous variables andχ2 test for categorical variables.In multivariate analysis,the odds ratio (OR )was calculated along with a 95 % confidence interval (CI )by binary Logistic regression. Results A total of 1 204 silica exposure patients had full basic information and 99.8% were male patients with mean age of (59.4 ± 6.8 )years.The patients in phase 0 + to phase Ⅲ were 172 (14.3%),255 (21 .2%),160 (13.3%)and 617 (51 .2%),respectively.The tuberculosis prevalence rate was about 7.3% among these population.The risk factors for tuberculosis including phase Ⅱ silicosis (OR =2.96, 95 %CI :1 .05 -8.32,P =0.04)and phase Ⅲ silicosis (OR=3.88,95 %CI :1 .58-9.56,P <0.01),and contacting with tuberculosis patients (OR=4.14,95 %CI :1 .91 -8.98,P <0.01).Patients complicated with tuberculosis lacked specific symptoms,but fever and weight loss were more frequent.Conclusion Tuberculosis is highly prevalent in silicotic patients,especially in patients with phase Ⅱ/Ⅲ silicosis and in patients with tuberculosis contact history.

Doxorubicin (DOX)belongs to anthraquinone anti-tumor antibiotics,which is now one of most widely used anti-cancer drugs in clinic,but it possesses cardiotoxicity.How to reduce its cardiotoxicity is still a hot problem in current research.This article made an overview for monitoring and prevention of DOX cardiotoxicity.