OBJECTIVE To investigate the mechanism of Qushi huoxue decoction in improving metabolic-associated fatty liver disease （MAFLD） based on the AMP-activated protein kinase/mammalian target of rapamycin/UNC-51-like kinase 1 （AMPK/mTOR/ULK1） autophagy signaling pathway.METHODS Male C57BL/6J mice were randomly divided into normal group， model group， and low-， medium-， and high-dose Qushi huoxue decoction groups [4.10， 8.20， 16.40 mg/（g·d）]， with 10 mice in each group. Except for the normal group， mice in the other groups were fed a high-fat diet for 12 weeks to establish the MAFLD model. Starting 24 hours after modeling， mice in each group were administered the drug solution or normal saline once daily for 6 consecutive weeks. Serum levels of total cholesterol （TC）， triglycerides （TG）， alanine transaminase （ALT）， aspartate transferase （AST）， low-density lipoprotein cholesterol （LDL-C）， and high-density lipoprotein cholesterol （HDL-C） were detected according to the kit instructions. Hematoxylin-eosin （HE） staining and oil red O staining were used to observe pathological changes in mouse liver tissue. Western blot and immunofluorescence staining were used to detect the expression and positive protein expression related to the AMPK/mTOR/ULK1 autophagy signaling pathway.RESULTS Compared with the normal group， the serum levels of TC， TG， ALT， AST， and LDL-C in the model group were significantly increased， while the HDL-C level was significantly decreased （ P ＜0.01）. HE staining showed obvious steatosis in hepatocytes of the model group， and oil red O staining showed a large amount of lipid droplet accumulation in liver tissue. Western blot and immunofluorescence staining results showed that the expression levels of p-mTOR/mTOR and p62 proteins， as well as the relative fluorescence intensities of p-mTOR and p62 proteins in liver tissue of the model group， were significantly increased/enhanced （ P ＜0.01）， while the expression levels of p-AMPK/AMPK， ULK1， and microtubule-associated protein 1 light chain 3Ⅱ （LC3Ⅱ） proteins， as well as the relative fluorescence intensities of p-AMPK and LC3 proteins， were significantly decreased/weakened （ P ＜0.01）. After intervention with Qushi huoxue decoction， the above pathological changes and abnormal indicators in liver tissue of mice in the low-， medium-， and high-dose groups were significantly reversed， with most indicators showing statistically significant differences （ P ＜0.05 or P ＜0.01）.CONCLUSIONS Qushi huoxue decoction can significantly improve liver pathology， blood lipid levels， and liver function in MAFLD model mice； this improvement may be achieved through the activation of the AMPK/mTOR/ULK1 autophagy signaling pathway.

As the important source of bone cells, osteoblasts are involved in bone formation and repair, and play a key role in maintaining bone balance. If the osteogenic differentiation process in vivo is disrupted, a variety of bone-related diseases may occur. Natural products, which have a wide range of sources, a wide variety of physiological activities, and few toxic side-effects, have been found in recent years to be able to regulate osteoblast differentiation. Based on the sources of natural products, this paper reviews the intervention of natural products from plant, animal and microbial sources on osteogenic differentiation, aiming to provide a theoretical basis for natural products in the treatment of bone diseases.

Andrographolide sulfonate (AS) is a sulfonated derivative of andrographolide extracted from Andrographis paniculata (Burm.f.) Nees, and has been approved for several decades in China. The present study aimed to investigate the novel therapeutic application and possible mechanisms of AS in the treatment of rheumatoid arthritis. Results indicated that administration of AS by injection or gavage significantly reduced the paw swelling, improved body weights, and attenuated pathological changes in joints of rats with adjuvant-induced arthritis. Additionally, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-1β in the serum and ankle joints were reduced. Bioinformatics analysis, along with the spleen index and measurements of IL-17 and IL-10 levels, suggested a potential relationship between AS and Th17 cells under arthritic conditions. In vitro, AS was shown to block Th17 cell differentiation, as evidenced by the reduced percentages of CD4⁺ IL-17A⁺ T cells and decreased expression levels of RORγt, IL-17A, IL-17F, IL-21, and IL-22, without affecting the cell viability and apoptosis. This effect was attributed to the limited glycolysis, as indicated by metabolomics analysis, reduced glucose uptake, and pH measurements. Further investigation revealed that AS might bind to hexokinase2 (HK2) to down-regulate the protein levels of HK2 but not glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or pyruvate kinase M2 (PKM2), and overexpression of HK2 reversed the inhibition of AS on Th17 cell differentiation. Furthermore, AS impaired the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signals in vivo and in vitro, which was abolished by the addition of lactate. In conclusion, AS significantly improved adjuvant-induced arthritis (AIA) in rats by inhibiting glycolysis-mediated activation of PI3K/AKT to restrain Th17 cell differentiation.
Animals ; Th17 Cells/immunology* ; Diterpenes/pharmacology* ; Arthritis, Rheumatoid/metabolism* ; Proto-Oncogene Proteins c-akt/immunology* ; Glycolysis/drug effects* ; Cell Differentiation/drug effects* ; Phosphatidylinositol 3-Kinases/genetics* ; Rats ; Male ; Rats, Sprague-Dawley ; Humans ; Andrographis paniculata/chemistry* ; Arthritis, Experimental/drug therapy* ; Interleukin-17/immunology* ; Signal Transduction/drug effects*

Objective:To learn about the epidemic characteristics of human brucellosis in Baoji City, and to identify brucellosis strains, so as to provide scientific basis for precise prevention and control of brucellosis in the city.Methods:A retrospective analysis method was used to conduct a descriptive epidemiological analysis of brucellosis cases in Baoji City reported in the China Disease Prevention and Control Information System from 2009 to 2022. Bacteriology culture, serum agglutination, biochemical test, phage cleavage and PCR were used to identify the isolates and nucleic acids, respectively.Results:From 2009 to 2022, a total of 314 cases of human brucellosis were reported in Baoji City, with an average annual incidence rate of 0.62/100 000, The incidence rate of different years was compared, the difference was statistically significant (χ 2 = 51 748 520.36, P < 0.001). The gender ratio of male to female was 3.30 ∶ 1.00 (241 ∶ 73). The age distribution ranged from 2 to 84 years old, with a median of 50 years old. The cases were concentrated in the 30 - 59 age group (206 cases, 65.61%), followed by the 60 - 84 age group (80 cases, 25.48%) and the 2 - 29 age group (28 cases, 8.92%). The peak of incidence was from April to August, with a total of 205 cases, accounting for 65.29%. Cases were reported in 213 townships, with the number of townships increasing from 1 in 2009 to 29 in 2022. The median time interval between onset and diagnosis of the case was 11 days. There was no significant difference in the time interval between onset and diagnosis among different genders, occupation and regions ( P > 0.05). A total of 10 strains of Brucella were isolated and identified as Brucella genus by BCSP31-PCR, and as Brucella melitensis by AMOS-PCR. After comprehensive identification, 4 strains were Brucella melitensis 3, 3 strains were mutant Brucella melitensis, and 3 strains were Brucella melitensis 1. Conclusion:The epidemic of brucellosis in Baoji City shows a fluctuating upward trend, with the epidemic scope gradually expanding, and the infected strains are all Brucella melitensis.

Objective To identify the multidimensional needs of postoperative gastric cancer patients for home-based nutritional management based on patient journey maps,and to provide a reference for carrying out nutritional management interventions.Methods Using descriptive qualitative research methods,we facilitated semi-structured in-depth interviews with 9 pairs of postoperative gastric cancer homebound patients and their primary caregivers from a tertiary general hospital in Yichang City,China,from September 2024 to January 2025,and analysed the data and drew the patient journey maps by content analysis.Results Totally 24 sub-themes were summarised from 4 aspects,namely tasks,emotions,pain points and opportunity points,and journey maps involving the acute recovery period,the transitional adaptation period and the nutritional reconstruction period were formed.Conclusion The nutritional needs of homebound patients after gastric cancer surgery are complex and variable,and their needs for dietary guidance,eating-related symptom management,and real-time counselling are highlighted.In the future,appropriate intervention strategies can be developed based on the journey maps to meet the multidimensional nutritional needs of patients.

Objective To identify the multidimensional needs of postoperative gastric cancer patients for home-based nutritional management based on patient journey maps,and to provide a reference for carrying out nutritional management interventions.Methods Using descriptive qualitative research methods,we facilitated semi-structured in-depth interviews with 9 pairs of postoperative gastric cancer homebound patients and their primary caregivers from a tertiary general hospital in Yichang City,China,from September 2024 to January 2025,and analysed the data and drew the patient journey maps by content analysis.Results Totally 24 sub-themes were summarised from 4 aspects,namely tasks,emotions,pain points and opportunity points,and journey maps involving the acute recovery period,the transitional adaptation period and the nutritional reconstruction period were formed.Conclusion The nutritional needs of homebound patients after gastric cancer surgery are complex and variable,and their needs for dietary guidance,eating-related symptom management,and real-time counselling are highlighted.In the future,appropriate intervention strategies can be developed based on the journey maps to meet the multidimensional nutritional needs of patients.

Objective:To learn about the epidemic characteristics of human brucellosis in Baoji City, and to identify brucellosis strains, so as to provide scientific basis for precise prevention and control of brucellosis in the city.Methods:A retrospective analysis method was used to conduct a descriptive epidemiological analysis of brucellosis cases in Baoji City reported in the China Disease Prevention and Control Information System from 2009 to 2022. Bacteriology culture, serum agglutination, biochemical test, phage cleavage and PCR were used to identify the isolates and nucleic acids, respectively.Results:From 2009 to 2022, a total of 314 cases of human brucellosis were reported in Baoji City, with an average annual incidence rate of 0.62/100 000, The incidence rate of different years was compared, the difference was statistically significant (χ 2 = 51 748 520.36, P < 0.001). The gender ratio of male to female was 3.30 ∶ 1.00 (241 ∶ 73). The age distribution ranged from 2 to 84 years old, with a median of 50 years old. The cases were concentrated in the 30 - 59 age group (206 cases, 65.61%), followed by the 60 - 84 age group (80 cases, 25.48%) and the 2 - 29 age group (28 cases, 8.92%). The peak of incidence was from April to August, with a total of 205 cases, accounting for 65.29%. Cases were reported in 213 townships, with the number of townships increasing from 1 in 2009 to 29 in 2022. The median time interval between onset and diagnosis of the case was 11 days. There was no significant difference in the time interval between onset and diagnosis among different genders, occupation and regions ( P > 0.05). A total of 10 strains of Brucella were isolated and identified as Brucella genus by BCSP31-PCR, and as Brucella melitensis by AMOS-PCR. After comprehensive identification, 4 strains were Brucella melitensis 3, 3 strains were mutant Brucella melitensis, and 3 strains were Brucella melitensis 1. Conclusion:The epidemic of brucellosis in Baoji City shows a fluctuating upward trend, with the epidemic scope gradually expanding, and the infected strains are all Brucella melitensis.

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 investigate and distinguish the useful index of recognizing blood culture contamination and bloodstream infection when the blood cultures were positive in clinical practice, and to help clinicians to recognize blood culture contamination.Methods:A retrospective study was conducted to analyze the clinical data and distribution of bacteria of blood culture from 303 patients with positive blood culture from January to December 2021 in Huashan Hospital, Fudan University. Age, distribution of departments, positive time of blood culture, neutrophil ratio, procalcitonin and C-reactive protein (CRP) were compared between bloodstream infection group and blood culture contamination group. Mann-Whitney U test and chi-square test were used for statistical comparisons. The factors for recognizing blood culture contamination were analyzed by multivariate logistic regression. Results:A total of 303 patients with positive blood culture were enrolled and divided into two groups, including 237 cases in bloodstream infection group and 66 cases in blood culture contamination group, with the contamination rate of 21.78%. There was statistically significant difference in the distribution of departments between the two groups ( χ2=11.28, P=0.024). The proportion of age≥60 years old (52.7%(125/237) vs 36.4%(24/66)), the neutrophil ratio(0.86(0.79, 0.91) vs 0.82(0.75, 0.88)), the levels of procalcitonin (1.05(0.23, 6.64) μg/L vs 0.17(0.11, 0.72) μg/L) and CRP (60.55(24.83, 132.83) mg/L vs 40.64(8.98, 95.83) mg/L) in the bloodstream infection group were higher than those in blood culture contamination group, and the differences were statistically significant ( χ2=5.54, U=9 523.00, 10 906.00 and 9 278.50, respectively, all P<0.05). Among the 303 patients with positive blood culture, 253 patients had a single positive blood culture, 41 patients had twice positive blood cultures and nine patients had three times positive blood cultures. The highest contamination rate of detected pathogen was coagulase-negative staphylococcus (54.72%(29/53)) in the patients with single positive blood culture. The positive time of blood culture in the bloodstream infection group was 3(3, 4) d, which was shorter than 4(3, 4) d in the blood culture contamination group, and the difference was statistically significant ( U=6 521.00, P=0.026). Multivariate logistic regression analysis showed that when the time to positivity ≤3 days and procalcitonin level ≥2.00 μg/L, the positive blood culture results were more likely to be bloodstream infection (odds ratio ( OR)=2.16, 1.96, respectively, both P<0.05). Conclusions:Bacteria associated with blood culture, time to positivity and clinical index all play important roles in recognizing blood culture contamination. When the time to positivity ≤3 days and procalcitonin level ≥2.00 μg/L, the positive blood culture results are more likely to be bloodstream infection.

Objective:To explore the heterogeneity among keloids before and after radiotherapy and identify the changes of key cell subpopulations and their interactions utilizing single cell RNA sequencing technology.Methods:Four patients provided a total of 12 samples, each consisting of keloid tissue before and after radiotherapy and the normal skin tissue adjacent to the untreated keloid. The keloid was divided into left and right sides from the midline, and the left-side keloid was fractionally irradiated with 20 Gy electron beam in total in 4 consecutive days. The right-side keloid was irradiated with 10 Gy in 2 fractions before surgery and 10 Gy in 2 fractions after surgery.Results:A total of 25 573 fibroblasts were analyzed and categorized into nine subgroups (fibroblasts 1-9). The proportion of fibroblast-2 increased after radiotherapy ( t=4.70, P<0.05). The number of classical monocytes and macrophages increased after radiotherapy, but there was no significant difference due to the shorter time of sample taking at 2 d after radiotherapy ( P>0.05). Macrophages (4 723 cells) were further divided into four categories. CellPhoneDB analysis showed that type-3 macrophages interacted significantly more closely with fibroblasts than type-1 and type-2 macrophages. The most prominent signaling pathways for the interactions between type-3 macrophages and major fibroblast subtypes were the collagen signaling pathway and the chemerin signaling pathway. These interactions were more pronounced in the keloid samples after radiotherapy. Conclusions:The interactions between type-3 macrophages and fibroblasts (such as fibroblast-2) may serve as an important point for future studies on radio-sensitization of keloids.