ObjectiveThis study aims to explore the mechanism and targets of Shenfu Injection in regulating glycolysis to intervene in myocardial fibrosis in chronic heart failure based on the hypoxia-inducible factor-1α （HIF-1α）/ 6-phosphofructo-2-kinase/fructose-2，6-bisphosphatase 3 （PFKFB3） signaling pathway. MethodsA rat model of chronic heart failure was established by subcutaneous injection of isoproterenol （ISO）. After successful modeling， the rats were randomly divided into the Sham group， Model group， Shenfu injection （SFI， 6 mL·kg^-1） group， and inhibitor （3PO， 35 mg·kg^-1） group， according to a random number table， and they were treated for 15 days. Cardiac function was evaluated by echocardiography， and serum N-terminal pro-brain natriuretic peptide （NT-proBNP） levels were detected by enzyme-linked immunosorbent assay （ELISA）. Fasting body weight and heart weight were measured， and the heart index （HI） was calculated. Pathological changes in myocardial tissue were observed by hematoxylin-eosin （HE） and Masson staining， and the fibrosis rate was calculated. Biochemical assays were used to determine serum levels of glucose （GLU）， lactic acid （LA）， and pyruvic acid （PA）. Western blot was used to analyze the expression of proteins related to the HIF-1α/PFKFB3 signaling pathway （HIF-1α and PFKFB3）， glycolysis-related proteins （HK1， HK2， PKM2， and LDHA）， and fibrosis-related proteins ［transforming growth factor （TGF）-β₁， α-smooth muscle actin （α-SMA）， and Collagen type Ⅰ α1 （ColⅠA1）］. Real-time PCR was used to detect the mRNA expression of HIF-1α and PFKFB3 in myocardial tissue. ResultsCompared with the Sham group， the Model group showed significantly decreased left ventricular ejection fraction （LVEF）， left ventricular shortening fraction （LVFS）， interventricular septal thickness （IVSd）， and interventricular septal strain （IVSs） （P<0.05）， while left ventricular internal dimension at end-diastole （LVDd） and end-systole （LVIDs） were increased （P<0.05）. Serum NT-proBNP levels were significantly increased （P<0.01）， and body weight was decreased. Heart weight was increased， and the HIT index was increased （P<0.05）. Myocardial tissue exhibited inflammatory cell infiltration and collagen fiber deposition， and the fibrosis rate was significantly increased （P<0.05）. Serum GLU was decreased （P<0.05）， while LA and PA levels were increased （P<0.05）. Protein expressions of HIF-1α， PFKFB3， HK1， HK2， PKM2， LDHA， TGF-β₁， α-SMA， and ColⅠA1， as well as the mRNA expression of HIF-1α and PFKFB3 were increased （P<0.05）. Compared with the Model group， both the SFI group and 3PO groups showed significant improvements in LVEF， LVFS， IVSd， and IVSs （P<0.05） and decreases in LVDd， LVIDs， and NT-proBNP levels （P<0.05）. Body weight was significantly increased. Heart weight was significantly decreased， and the HIT index was significantly decreased （P<0.05）. Inflammatory cell infiltration， collagen fiber deposition， and the fibrosis rate were significantly decreased （P<0.05）. Serum GLU levels were significantly increased （P<0.05）， while LA and PA levels were decreased （P<0.05）. Expressions of glycolysis-related proteins， fibrosis-related proteins， and HIF-1α/PFKFB3 pathway-related proteins and mRNAs were significantly suppressed （P<0.05）. ConclusionSFI improves cardiac function in chronic heart failure by downregulating the expression of HIF-1α/PFKFB3 signaling pathway-related proteins， regulating glycolysis， and inhibiting myocardial fibrosis.

Objective:To investigate effect of artesunate(ART)on high glucose(HG)induced trophoblast cell damage and its mechanism.Methods:Human trophoblast cells HTR-8/SVneo were separated into control group,HG group,ART low concentra-tion group,ART high concentration group and ART high concentration+lysophosphatidic acid(LPA)group.CCK-8 method was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis.ELISA was applied to detect levels of TNF-α,IL-1β and IL-10.Reagent kits were used to detect levels of lactate dehydrogenase(LDH),malondialdehyde(MDA)and superoxide dismutase(SOD).Western blot was applied to detect expressions of Bax,Bcl-2,Ras homolog gene family member A(RhoA),Rho associated coiled coil containing protein kinase(ROCK)1 and ROCK2.Results:After treatment with 0.4～1.6 μmol/L ART,activity of HTR-8/SVneo cells were increased(P<0.05).ART greatly enhanced activity of HTR-8/SVneo cells induced by HG,and cell viability was most significantly increased under 1.6 μmol/L ART treatment(P<0.05).Compared with control group,activity of HTR-8/SVneo cells,IL-10,SOD and Bcl-2 levels were reduced in HG group,apoptosis rate,TNF-α,IL-1β,LDH,MDA,Bcl-2,RhoA,ROCK1 and ROCK2 levels were increased(P<0.05).Compared with HG group,activity of HTR-8/SVneo cells,IL-10,SOD and Bcl-2 levels were increased in ART low and high concentration groups,apoptosis rate,TNF-α,IL-1β,LDH,MDA,Bcl-2,RhoA,ROCK1 and ROCK2 levels were decreased(P<0.05).Compared with ART high concentration group,activity of HTR-8/SVneo cells,IL-10,SOD and Bcl-2 levels were decreased in ART high concentration+LPA group,apoptosis rate,TNF-α,IL-1β,LDH,MDA,Bcl-2,RhoA,ROCK1 and ROCK2 levels were increased(P<0.05).Conclusion:ART can inhibit inflammatory response and oxidative stress of trophoblast cells and reduce cell apoptosis,whose mechanism may be related to inhibition of RhoA/ROCK signaling pathway.

Objective To investigate the effects of adiponectin(APN)on the proliferation,migration,and invasion of ovarian cancer cells and its potential association with the phosphatidylinositol 3-kinase(PI3K)/pro-tein kinase B(Akt)/mammalian target of rapamycin(mTOR)signaling pathway.Methods A2780 and SK-OV3 cell lines were selected.AdipoR1 mRNA expression was detected by real-time polymerase chain reaction(RT-PCR).The CCK-8 assay was employed to evaluate APN-induced proliferation,while scratch wound heal-ing and Transwell assays were conducted to assess migration and invasion capabilities.Western blot analysis was performed to measure the expression levels of PI3K,Akt,mTOR,and their phosphorylated forms(p-Akt,p-mTOR)at different time points(0-60 min)after APN treatment and across intervention groups(APN,LY294002,APN+LY294002).Results Both A2780 and SKOV3 cells expressed AdipoR1 mRNA.Compared with the control group,APN significantly enhanced the proliferation,migration,and invasion of o-varian cancer cells(P＜0.05).Under the stimulation of APN,the expression levels of p-Akt and p-mTOR in A2780 and SKOV3 cells showed a significant upward trend.The peak expression of p-Akt and p-mTOR in A2780 cells was observed at 30 minutes,while in SKOV3 cells,it occurred at 60 minutes.The expression lev-els of p-Akt and p-mTOR were significantly reduced in the LY294002 group and APN+LY294002 group(P＜0.001).Conclusion APN enhances the proliferation,migration,and invasion behavior of ovarian cancer cells by activating the PI3K/Akt/mTOR signaling pathway,and its effect can be reversed by PI3K inhibitors,providing a theoretical basis for targeted intervention in ovarian cancer progression.

Objective:To investigate effect of artesunate(ART)on high glucose(HG)induced trophoblast cell damage and its mechanism.Methods:Human trophoblast cells HTR-8/SVneo were separated into control group,HG group,ART low concentra-tion group,ART high concentration group and ART high concentration+lysophosphatidic acid(LPA)group.CCK-8 method was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis.ELISA was applied to detect levels of TNF-α,IL-1β and IL-10.Reagent kits were used to detect levels of lactate dehydrogenase(LDH),malondialdehyde(MDA)and superoxide dismutase(SOD).Western blot was applied to detect expressions of Bax,Bcl-2,Ras homolog gene family member A(RhoA),Rho associated coiled coil containing protein kinase(ROCK)1 and ROCK2.Results:After treatment with 0.4～1.6 μmol/L ART,activity of HTR-8/SVneo cells were increased(P<0.05).ART greatly enhanced activity of HTR-8/SVneo cells induced by HG,and cell viability was most significantly increased under 1.6 μmol/L ART treatment(P<0.05).Compared with control group,activity of HTR-8/SVneo cells,IL-10,SOD and Bcl-2 levels were reduced in HG group,apoptosis rate,TNF-α,IL-1β,LDH,MDA,Bcl-2,RhoA,ROCK1 and ROCK2 levels were increased(P<0.05).Compared with HG group,activity of HTR-8/SVneo cells,IL-10,SOD and Bcl-2 levels were increased in ART low and high concentration groups,apoptosis rate,TNF-α,IL-1β,LDH,MDA,Bcl-2,RhoA,ROCK1 and ROCK2 levels were decreased(P<0.05).Compared with ART high concentration group,activity of HTR-8/SVneo cells,IL-10,SOD and Bcl-2 levels were decreased in ART high concentration+LPA group,apoptosis rate,TNF-α,IL-1β,LDH,MDA,Bcl-2,RhoA,ROCK1 and ROCK2 levels were increased(P<0.05).Conclusion:ART can inhibit inflammatory response and oxidative stress of trophoblast cells and reduce cell apoptosis,whose mechanism may be related to inhibition of RhoA/ROCK signaling pathway.

Objective:To explore facilitators and impediments to home glucose management for patients undergoing bariatric surgery based on the consolidated framework for implementation research (CFIR), so as to inform the development of a glucose management pathway after bariatric surgery.Methods:This study was a descriptive phenomenological study. Using purposive sampling method, seven healthcare professionals and 11 patients undergoing bariatric surgery from the General Surgery·Metabolic and Bariatric Center of China-Japan Friendship Hospital were selected from November 2023 to January 2024 for semi-structured interviews. Targeted content analysis was used for the data analysis.Results:A total of six facilitators and six impediments related to the five dimensions of CFIR were distilled. Facilitators included the personalized glucose management program, regular long-term follow-up, multidisciplinary management team, support from family and friends, complication distress, and information technology platform. Impediments involved patterned follow-up, poor accessibility and convenience of resources, role conflict, herd mentality, poor behavioral compliance, and low utilization of digital equipment.Conclusions:Glucose management needs vary in patients after bariatric surgery, with a gradual decline in management compliance. Healthcare professionals should build an "Internet +"home glucose management service model, carry out targeted multidisciplinary team follow-up, and promote the formation of a standardized pathway for patients' home glucose management.

Objective:To explore facilitators and impediments to home glucose management for patients undergoing bariatric surgery based on the consolidated framework for implementation research (CFIR), so as to inform the development of a glucose management pathway after bariatric surgery.Methods:This study was a descriptive phenomenological study. Using purposive sampling method, seven healthcare professionals and 11 patients undergoing bariatric surgery from the General Surgery·Metabolic and Bariatric Center of China-Japan Friendship Hospital were selected from November 2023 to January 2024 for semi-structured interviews. Targeted content analysis was used for the data analysis.Results:A total of six facilitators and six impediments related to the five dimensions of CFIR were distilled. Facilitators included the personalized glucose management program, regular long-term follow-up, multidisciplinary management team, support from family and friends, complication distress, and information technology platform. Impediments involved patterned follow-up, poor accessibility and convenience of resources, role conflict, herd mentality, poor behavioral compliance, and low utilization of digital equipment.Conclusions:Glucose management needs vary in patients after bariatric surgery, with a gradual decline in management compliance. Healthcare professionals should build an "Internet +"home glucose management service model, carry out targeted multidisciplinary team follow-up, and promote the formation of a standardized pathway for patients' home glucose management.

Interactions between brain-resident and peripheral infiltrated immune cells are thought to contribute to neuroplasticity after cerebral ischemia. However, conventional bulk sequencing makes it challenging to depict this complex immune network. Using single-cell RNA sequencing, we mapped compositional and transcriptional features of peri-infarct immune cells. Microglia were the predominant cell type in the peri-infarct region, displaying a more diverse activation pattern than the typical pro- and anti-inflammatory state, with axon tract-associated microglia (ATMs) being associated with neuronal regeneration. Trajectory inference suggested that infiltrated monocyte-derived macrophages (MDMs) exhibited a gradual fate trajectory transition to activated MDMs. Inter-cellular crosstalk between MDMs and microglia orchestrated anti-inflammatory and repair-promoting microglia phenotypes and promoted post-stroke neurogenesis, with SOX2 and related Akt/CREB signaling as the underlying mechanisms. This description of the brain's immune landscape and its relationship with neurogenesis provides new insight into promoting neural repair by regulating neuroinflammatory responses.
Humans ; Ischemic Stroke ; Brain/metabolism* ; Macrophages ; Brain Ischemia/metabolism* ; Microglia/metabolism* ; Gene Expression Profiling ; Anti-Inflammatory Agents ; Neuronal Plasticity/physiology* ; Infarction/metabolism*

Objective:To explore the differences of gene expression profiles of precursors of exhausted T cells (Tpex) and terminal exhausted T cells (Tex) in the peripheral blood of patients with active pulmonary tuberculosis (ATB).Methods:Twenty-five cases of ATB, 13 cases of latent tuberculosis infection (LTBI) and 10 health controls were enrolled from January 2021 to October 2022 in the Fifth People′s Hospital of Wuxi. The proportions of Tpex and Tex in the peripheral blood mononuclear cells (PBMCs) of the three groups were detected by flowcytometry. PBMCs of ATB were separated into Tpex and Tex by fluorescence-activated cell sorting. RNA-sequencing was performed and up-regulated and down-regulated genes were screended. Differently expressed genes were analyzed by gene set enrichment analysis of gene ontology (GO) to find regulatory pathways affecting cell metabolism and function. Wilcoxon matched-pairs signed rank test, Kruskal-Wallis test and Dunn multiple comparsion test were used for statistical analysis.Results:The proportion of Tpex in ATB group was 2.86%(1.74%), which was lower than 7.93%(6.16%) of Tex, and the difference was statistically significant ( Z=-3.91, P<0.001). The proportions of Tpex and Tex in LTBI group were 9.47%(6.26%) and 7.43%(5.48%), respectively, and the difference was not statistically significant ( Z=-0.93, P=0.345). The proportions of Tpex and Tex in healthy control group were 8.42%(2.69%) and 6.49%(5.14%), respectively, with no statistical significance ( Z=-1.36, P=0.170). There was statistical difference of the proportion of Tpex among the three groups ( H=21.93, P<0.001), and the proportion of Tpex in ATB group was lower than those in LTBI and heathy control groups, and the differences were both statistically significant ( Z=4.16, P<0.001 and Z=3.34, P=0.003, respectively), while the proportions of Tex in these three groups were not statistically different ( H=2.17, P=0.338). Compared with Tex, the gene expressions of memory markers, such as B-cell lymphoma 2 of Tpex were up-regulated, and the gene expressions of exhausted markers, such as lymphocyte activation gene 3 were down-regulated. In terms of cellular metabolism, the gene expressions of mitochondrial protein complex, mitochondrial matrix and oxidative phosphorylation of Tpex were up-regulated, and the gene expressions of glycolysis were down-regulated. The gene expressions of pyruvate metabolism in Tex were up-regulated, and the gene expressions of CD4 + T lymphocyte activation and differentiation and glycolytic process in Tpex were down-regulated. Conclusions:Tpex in ATB express more characteristics of memory cells and less features of exhausted markers compared with Tex, and the function of mitochondria of Tpex preserves well.

Objective:To investigate the clinical features of nontuberculous mycobacteria (NTM) disease patients with positive anti-interferon γ (IFN-γ) autoantibody.Methods:Forty-three adult human immunodeficiency virus-uninfected patients with NTM disease hospitalized in Huashan Hospital, Fudan University and Jing′an Branch, Huashan Hospital, Fudan University from July 2021 to August 2023 were included. Clinical data and NTM strain information of the patients were collected. The plasma levels of anti-IFN-γ autoantibodies were detected by enzyme-linked immunosorbent assay, and the patients were divided into antibody positive group and antibody negative group. The clinical characteristics and laboratory examination results between the two groups were compared. The independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Multivariate logistic regression analysis was used to determine the correlation factors of positive anti-IFN-γ autoantibodies. Results:Among the 43 patients, 13 cases (30.2%) were positive for anti-IFN-γ autoantibodies and 30 cases (69.8%) were negative. The proportions of patients with NTM disseminated infection (9/13 vs 30.0%(9/30))and combined bacterial infection (5/13 vs 6.7%(2/30)) in antibody positive group were both higher than those in antibody negative group, and the differences were both statistically significant ( χ2=5.74 and 6.73, respectively, both P<0.05). The white blood cell count, platelet count, the proportion of platelet count >350×10 9/L of antibody positive patients were all higher than those of antibody negative group, while the white sphere ratio was lower than that of antibody negative group, with statistical significance ( t=2.42, 3.02, χ2=9.77 and t=3.66, respectively, all P<0.05). Erythrocyte sedimentation rate, C-reactive protein, procalcitonin, globulin, immunoglobulin G, immunoglobulin A and immunoglobulin M in antibody positive patients were all higher than those in antibody negative group, and the differences were all statistically significant ( U=99.50, 112.00, 115.50, 61.50, 76.50, 99.00 and 83.00, respectively, all P<0.05). Mycobacterium abscessus complex (seven cases and 11 cases, respectively) and Mycobacterium avium complex (five cases and 13 cases, respectively) were the main isolated strains in antibody positive and antibody negative patients. Multivariate logistic regression analysis showed that combined with bacterial infection (odds ratio ( OR)=21.83, 95% confidence interval ( CI) 1.94 to 245.71), NTM disseminated infection ( OR=7.64, 95% CI 1.10 to 53.26), platelet count>350×10 9/L ( OR=14.31, 95% CI 1.91 to 107.04) were risk factors for anti-IFN-γ autoantibodies positive (all P<0.05). Conclusions:Patients with positive anti-IFN-γ autoantibodies have higher probability of having elevated levels of systemic inflammation. Anti-IFN-γ autoantibody test is recommended for patients with NTM disease who present with co-bacterial infection, NTM disseminated infection, or elevated platelet count (>350×10 9/L).

Objective To translate the Eating Behaviors Assessment for Obesity(EBA-O)and test its reliability and validity for providing an effective tool for brief and rapid clinical assessment of dietary behavior in obese patients.Methods The Chinese version of EBA-O was developed by Brislin's translation model for translation,back translation,cultural adaptation,and pilot investigation.Using convenience sampling method,200 obese patients who were admitted to the inpatient ward of the Weight Loss Center of a tertiary hospital in Beijing from April to June 2023 and planned to undergo weight loss metabolic surgery were selected as the research participants to test the reliability and validity of the Chinese version of EBA-O.Results The Chinese version of EBA-O includes 4 dimensions,with a total of 17 items.The Cronbach's α coefficient of the scale is 0.886;the Cronbach's αcoefficients of the dimensions are 0.770～0.866;the test-retest reliability of the scale is 0.881.The content validity index of the scale level is 0.980,and the content validity indexes of item level were 0.889～1.000.Totally 4 common factors were extracted through exploratory factor analysis,with a cumulative variance of 66.363％.Conclusion The Chinese version of EBA-O has good reliability and validity,and it can be used as a tool to evaluate the dietary behavior of obese patients undergoing weight loss metabolic surgery.