ObjectiveEndothelial cell dysfunction being the core link. This study explores the molecular mechanism of Danshen Tongluo formula in treating coronary artery disease-dominated panvascular disease with endothelial cell changes as the core through animal experiments and single-cell transcriptome sequencing. MethodsA rat model of coronary artery disease-dominated panvascular disease was established by ligating the left anterior coronary artery. Rats were randomized into a blank group， a model group， and a Danshen Tongluo formula （28 mg·kg^-1·d^-1） group. The efficacy was evaluated by examining the cardiac ultrasound， determination of the plasma level of N-terminal pro-brain natriuretic peptide， and pathological staining. After single-cell sequencing， SingleR package， public datasets， and related literature were used for annotation of the cells. Cell chat was used for intercellular communication and ligand-receptor analysis， and scmetabolism was used for metabolic analysis of endothelial cells. ResultsAnimal experiments showed that Danshen Tongluo formula reduced the N-terminal pro-brain natriuretic peptide （ NT-proBNP ） level （P<0.05）， ameliorated myocardial cell damage and fibrosis， and increase left ventricular ejection fractions （LVEF） in the rat model of heart failure after myocardial infarction（P<0.05）. Single-cell sequencing results showed that Danshen Tongluo formula increased the proportion of arterial endothelial cells， venous endothelial cells， and capillary-arterial endothelial cells， while reducing the proportion of capillary-venous endothelial cells. In addition， this formula increased the interaction intensity of endothelial cells with cardiomyocytes and M1 macrophages and reduced the interaction intensity of endothelial cells with fibroblasts and T cells. Danshen Tongluo formula upregulated CXCL12-CXCR4 signaling in endothelium-B cells and Ptprm-Ptprm signaling in endothelial endothelial cells， while downregulating Mif-（CD74⁺CXCR44） signaling in endothelium-M1 macrophages and Mif-（CD74⁺CD44） signaling in endothelium-M2 macrophages. It reduced the citric acid cycle， oxidative phosphorylation， and glycolysis and increased the glycolysis/oxidative phosphorylation ratio in endothelial cells. GO and KEGG enrichment analysis showed that arterial endothelial cells， venous endothelial cells， and venous capillary endothelial cells can all regulate oxidative phosphorylation， cell adhesion molecules， and tyrosine metabolism. Lymphatic endothelial cells regulate immunity and vascular constriction to participate in the metabolism of various amino acids and fatty acids. ConclusionDanshen Tongluo Formula can ameliorate coronary artery disease-dominated panvascular disease by changing the composition of endothelial cells and regulating the communication between myocardial endothelial cells and non-endothelial cells.

Objective To explore the efficacy of the self-formulated Zhuzhang Formula in trea-ting the spleen deficiency with dysfunction of transportation syndrome of idiopathic short stature(ISS)and its effects on the growth rate,disease-related indicators,and bone metabolism indicators in affect-ed children.Methods A total of 80 children with ISS were enrolled as study subjects.They were randomly assigned to control group and observation group using random number table method,with 40 cases in each group.The control group received conventional western medicine treatment,while the observation group was administered self-formulated Zhuzhang Formula in addition to the treatment of the control group.Comparisons were made between the two groups in terms of clinical efficacy,TCM syndrome scores,growth parameters(monthly average height increment,annual growth rate),levels of disease-related indicators[serum insulin-like growth factor-1(IGF-1),bone morphogenetic pro-tein-2(BMP-2),insulin-like growth factor binding protein-3(IGFBP-3)],levels of bone metabo-lism indicators[bone alkaline phosphatase(BAP),type Ⅰ procollagen N-terminal propeptide(PⅠNP),type Ⅰ collagen cross-linked C-terminal telopeptide(β-CTX)],and the incidence of adverse reac-tions.Results The overall effective rate in the observation group was 97.50％,which was significantly higher than the 80.00％ in the control group(P＜0.05).After 3 and 6 months of intervention,the scores for symptoms such as short stature,sallow complexion,loss of appetite,weak voice and shortness of breath,limb weakness,and loose stools,as well as the total TCM syndrome scores,were lower in the observation group than those in the control group,with statistically significant differences(P＜0.05).After 6 months of intervention,the monthly average height increment and annual growth rate were higher in the observation group than those in the control group,and the differences were statistically significant(P＜0.05).After 3 and 6 months of intervention,the levels of IGF-1,BMP-2,IGFBP-3,BAP,and PⅠNP were higher in the observation group than those in the control group,while the level of β-CTX was lower,and the differences were statistically significant(P＜0.05).There was no statistically significant difference in the incidence of adverse reactions between the two groups(P＞0.05).Conclusion The self-formulated Zhuchang Formula demon-strates remarkable efficacy in treating the spleen deficiency with dysfunction of transportation syndrome of ISS.It can alleviate clinical symptoms by regulating the levels of IGF-1,BMP-2,IGFBP-3,and bone metabolism indicators,thereby promoting the growth and development of affected children,and exhibits high safety.

Objective To analyze the impact of sarcopenia on the efficacy and adverse reactions of immunotherapy combined with chemotherapy in patients with advanced gastric cancer.Methods Patients with locally advanced or metastatic gastric cancer confirmed by pathology who were not eligible for radical surgery were selected as study subjects.A body composition analyzer was used to measure the appendicular muscle mass of the patients and calculate the skeletal muscle mass index(SMI).Based on the SMI,the patients were divided into sarcopenia group and non-sarcopenia group.On the basis of nutritional intervention and comprehensive exercise therapy,the patients were administered immu-notherapy combined with chemotherapy.The efficacy and adverse reactions were evaluated.The primary endpoint was progression-free survival(PFS),and the secondary endpoints were the objec-tive response rate(ORR)and treatment-related adverse reactions.Results A total of 52 gastric cancer patients were included,with 23 in the sarcopenia group and 29 in the non-sarcopenia group.The median PFS in the non-sarcopenia group was 9.8 months(95％CI,8.9 to 12.4),and was 5.4 months in the sarcopenia group(95％CI,4.9 to 8.1).The median PFS in the non-sarcopenia group was longer than that in the sarcopenia group,and the difference was statistically significant[HR(95％CI)=0.41(0.23 to 0.73),P=0.003].The results of the multivariate Cox propor-tional hazards regression model showed that comorbidities,treatment cycles,and sarcopenia were all independent prognostic factors affecting the PFS of gastric cancer patients(P＜0.05).The ORR in the non-sarcopenia group was 48.28％(14/29),and was 17.39％(4/23)in the sarcopenia group(x2=5.276,P＜0.05).Treatment-related adverse reactions with grading ≥3 in both groups were mainly hematological toxicities.In the non-sarcopenia group,the incidence of grading ≥ 3 treat-ment-related adverse reactions was 27.59％(8/29),and the incidence of grading＜3 treatment-re-lated adverse reactions(including those with no adverse reactions)was 72.41％(21/29).In the sarcopenia group,the incidence of grading ≥3 treatment-related adverse reactions was 56.52％(13/23),and the incidence of grading＜3 treatment-related adverse reactions(including those without adverse reactions)was 43.48％(10/23).The incidence of grading ≥3 treatment-related adverse reactions in the non-sarcopenia group was lower than that in the sarcopenia group(P=0.035).Conclusion For patients with locally advanced or metastatic gastric cancer complicated with sarcopenia,the median PFS of immunotherapy combined with chemotherapy is shorter,the ORR is lower,and the incidence of treatment-related adverse reactions is increased.Therefore,ear-ly intervention for sarcopenia should be implemented to improve the quality of life of patients with advanced gastric cancer.

Objective:To explore the causal relationship between thyroid dysfunction and sepsis based on the bidirectional two-sample Mendelian randomization (MR) method.Methods:The genome-wide association study (GWAS) dataset were selected to screen single nucleotide polymorphisms (SNP) associated with thyroid dysfunction as instrumental variable (IV) for genetic variation, using hypothyroidism and hyperthyroidism as exposure factor and sepsis as outcome factor. Potential causal relationship between thyroid dysfunction and sepsis was analyzed using a bidirectional two-sample MR method primary analysis method of inverse-variance weighted (IVW). Potential pleiotropic analysis of SNP was performed using the MR Egger regression intercept test. Sensitivity analysis was performed using the "leave one out" test. Reverse MR method was used to prove the causal relationship.Results:The GWAS data were screened based on the three main assumptions of MR, resulting in 101 SNP strongly associated with hypothyroidism and 10 SNP strongly associated with hyperthyroidism entering the MR analysis. The results of the MR using the IVW method showed that the risk of sepsis in individuals with hypothyroidism was 2.293 times higher than those without hypothyroidism [odds ratio ( OR) = 2.293, 95% confidence interval (95% CI) was 1.199-4.382, P = 0.012]. There was no significant difference in the risk of sepsis between hyperthyroid and non-hyperthyroid populations ( OR = 1.049, 95% CI was 0.999-1.100, P = 0.560). MR Egger regression intercept test showed that the included SNP did not have pleiotropy, and the MR-PRESSO test did not find outliers. Sensitivity analysis suggested that the results of MR were stable. The results of the reverse MR analysis showed that the reverse causal relationship between hyperthyroidism and sepsis was not proved ( OR = 0.996, 95% CI was 0.988-1.004, P = 0.338), which further confirmed the robust MR analysis result. Conclusion:The results of the bidirectional two-sample MR analysis show that hypothyroidism can increase the risk of sepsis onset, while there is no causal relationship between hyperthyroidism and sepsis.

Objective To observe the effects of Huoxue Bushen Anshen Prescription on energy metabolism of H9c2 cardiomyocytes with oxidative stress injury.Methods H9c2 cardiomyocytes oxidative stress model was established with H2O2 to simulate myocardial injury.CCK-8 method was used to select the best conditions for modeling and the appropriate concentration of drugs.Cells were divided into normal group,model group,trimetazidine hydrochloride group and Huoxue Bushen Anshen Prescription group.Na+-K+-ATPase activity,real-time ATP generation rate,mitochondrial pressure,gycolysis rate,and long-chain fatty acid oxidationl pressure on H9c2 cardiomyocytes after treatment were performed,and the changes in energy metabolism of cardiomyocytes were observed.Results Compared with the normal group,the Na+-K+-ATPase activity,glycolytic ATP production rate,mitochondrial ATP production rate,basal oxygen consumption,non mitochondrial oxygen consumption,ATP production capacity,cellular reserve respiratory capacity and maximum respiratory capacity of H9c2 cells in the model group decreased,the glycolytic rate slowed down,long chain fatty acid basal respiration,acute response and maximum respiratory value decreased(P<0.05,P<0.01).Compared with the model group,the H9c2 cells in Huoxue Bushen Anshen Prescription group showed an increase in Na+-K+-ATPase activity,an increase in glycolytic ATP production rate and mitochondrial ATP production rate,an increase in basal oxygen consumption,non mitochondrial oxygen consumption,ATP production capacity,cellular reserve respiratory capacity and maximum respiratory capacity,an increase in basal glycolysis rate,and an increase in long-chain fatty acid basal respiration,acute response and maximum respiratory value(P<0.05,P<0.01).Conclusion Huoxue Bushen Anshen Prescription can significantly improve the activity of Na+-K+-ATPase in cH9c2 ardiomyocytes with oxidative stress injury,increase the rate of ATP production,improve mitochondrial pressure,glycolysis rate and oxidation pressure of long chain fatty acids,and improve the energy metabolism of cardiomyocytes.

Objective To explore the mechanism of Huoxue Bushen Anshen Prescription in improving the oxidative damage of H9c2 cells based on AhR-JDP2-Nrf2 axis.Methods H9c2 cells were divided into normal group,model group,TCM group,trimetazidine group,empty plasmid group,overexpression group,TCM+overexpression group and trimetazidine+overexpression group.The oxidative stress model of H9c2 cells was induced with H2O2 to imitate myocardial injury,and the cells were transfected with JDP2 overexpressed gene,intervened with Huoxue Bushen Anshen Prescription and trimetazidine hydrochloride respectively.Apoptosis was detected by flow cytometry,the contents of MDA,SOD and NADPH in cell supernatant were detected by kits,the protein expression of AhR,JDP2,Nrf2 and HO-1 were detected by Western blot,the mRNA expression of AhR,JDP2,Nrf2 and HO-1 were detected by RT-qPCR.Results Compared with the normal group,the early apoptosis rate,late apoptosis rate and total apoptosis rate of H9c2 cells in the model group increased(P<0.01),the contents of MDA,SOD and NADPH in cell supernatant increased(P<0.05),the expression of AhR,JDP2,Nrf2,HO-1 protein and mRNA increased(P<0.05).Compared with the model group,the early apoptosis rate,late apoptosis rate and total apoptosis rate of H9c2 cells in the TCM group and the trimetazidine group significantly decreased(P<0.01),the contents of MDA,SOD and NADPH decreased(P<0.05),the expression of AhR,JDP2,Nrf2 and HO-1 protein and mRNA decreased in the TCM group(P<0.05).Compared with the empty plasmid group,the overexpression group showed an increase in early apoptosis rate,late apoptosis rate and total apoptosis rate of H9c2 cells(P<0.01),an increase in MDA and NADPH content in cell supernatant(P<0.01),an decrease in SOD content(P<0.01),and an increase in the expressions of AhR,JDP2,Nrf2,HO-1 protein and mRNA in H9c2 cells(P<0.05).Compared with the overexpression group,the early apoptosis rate,late apoptosis rate and total apoptosis rate of H9c2 cells in the TCM+overexpression group and the trimetazidine+overexpression group were significantly decreased(P<0.05,P<0.01),the MDA and NADPH contents in cell supernatant were decreased(P<0.01),the SOD content increased(P<0.01),and the expressions of AhR,JDP2,Nrf2,HO-1 protein and mRNA in H9c2 cells decreased(P<0.05).Conclusion Huoxue Bushen Anshen Prescription can regulate the AhR-JDP2-Nrf2 axis by regulating the expression of JDP2,alleviate the oxidative damage of H9c2 cells,reduce the apoptosis of cardiomyocytes,and protect cardiomyocytes.

Objective To analyze the effects of massage therapy on the inflammatory state and lung function of pediatric refractory Mycoplasma pneumonia (RMPP). Methods A total of 320 children with RMPP treated in Hebei Seventh People′s Hospital from September 2022 to August 2023 were selected in the study. According to different treatment methods, they were divided into two groups, with 160 cases in each group. The western medicine group was given conventional anti-inflammatory and other symptomatic treatment, while the massage group was given dialectical massage treatment based on the western medicine group. The clinical efficacy, duration of disappearance of clinical symptoms, pulmonary function indexes, serum inflammatory factors, immune function indexes and total incidence of adverse reactions were compared between the two groups. Results The total effective rate of massage group was higher than that of control group (P < 0.05). The disappearance time of fever, cough, sputum and lung moist rales in observation group was shorter than that in control group (P < 0.05). After treatment, the peak expiratory flow (PEF), maximum mid-expiratory flow (MMEF) and forced vital capacity (FVC) in the massage group were higher than those in the control group (P < 0.05). The levels of serum C-reactive protein (CRP), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in the massage group were lower than those in the control group after treatment (P < 0.05). After treatment, CD3⁺, CD4⁺, CD4⁺/CD8⁺ in the massage group were higher than those in the control group (P < 0.05), and CD8⁺ was lower than that in the control group (P < 0.05). There was nosignificant difference in the total incidence of adverse reactions between the massage group and the western medicine group (P>0.05). Conclusion Massage based on syndrome differentiation combined with western medicine can effectively relieve fever, cough and other symptoms of RMPP children, reduce inflammatory response, improve lung function and immune function, and has less adverse reactions.

Background: Ferroptosis, which is caused by an iron-dependent accumulation of lipid hydroperoxides, is a type of cell death linked to diabetic kidney disease (DKD). Previous research has shown that fatty acid binding protein 4 (FABP4) is involved in the regulation of ferroptosis in diabetic retinopathy. The present study was constructed to explore the role of FABP4 in the regulation of ferroptosis in DKD. Methods: We first detected the expression of FABP4 and proteins related to ferroptosis in renal biopsies of patients with DKD. Then, we used a FABP4 inhibitor and small interfering RNA to investigate the role of FABP4 in ferroptosis induced by high glucose in human renal proximal tubular epithelial (HG-HK2) cells. Results: In kidney biopsies of DKD patients, the expression of FABP4 was elevated, whereas carnitine palmitoyltransferase-1A (CP-T1A), glutathione peroxidase 4, ferritin heavy chain, and ferritin light chain showed reduced expression. In HG-HK2 cells, the induction of ferroptosis was accompanied by an increase in FABP4. Inhibition of FABP4 in HG-HK2 cells changed the redox state, sup-pressing the production of reactive oxygen species, ferrous iron (Fe2+), and malondialdehyde, increasing superoxide dismutase, and reversing ferroptosis-associated mitochondrial damage. The inhibition of FABP4 also increased the expression of CPT1A, reversed lipid deposition, and restored impaired fatty acid β-oxidation. In addition, the inhibition of CPT1A could induce ferroptosis in HK2 cells. Conclusion Our results suggest that FABP4 mediates ferroptosis in HG-HK2 cells by inhibiting fatty acid β-oxidation.

OBJECTIVE To optimize the heart-nourishing granule extraction process by network pharmacology and Box-Behnken response surface method. METHODS The active ingredients of heart-nourishing granule were excavated through network pharmacology and their mechanism of action in the treatment of coronary heart disease was preliminarily explored. Molecular docking technology was used to predict the binding ability of the active ingredients to the main targets. At the same time, based on the compatibility relationship between the monarch, minister, assistant and guide of the prescription, the pharmacological effects of the ingredients, and the content determination index components of each medicinal flavor in the 2020 edition of the Chinese Pharmacopoeia, the process evaluation index components of heart-nourishing granule were further determined. In addition, combined with the analytic hierarchy process to determine the weight of each component, the Box-Behnken response surface method was used to optimize the extraction process of heart-nourishing granule. RESULTS The main active components of heart-nourishing granule screened were catalpol, Rhmannioside D, acteoside, ferulic acid and lobetyolin. By acting on core targets such as AKT1, IL-6, IL-1b, VEGFA, JUN and MAPK3, they regulated lipid and atherosclerosis, MAPK signaling and other pathways played a role in the treatment of coronary heart disease. Molecular docking results showed that the binding energies of active components and core targets predicted by network pharmacology were all < -5.0 kJ·mol-1. Five components catalpol, Rhmannioside D, acteoside, ferulic acid and lobetyolin were calculated by analytic hierarchy process method. The weight coefficients of arginyl glycosides were 0.329 7, 0.329 7, 0.164 8, 0.109 9, and 0.065 9, respectively. The Box-Behnken response surface method obtained the optimal water extraction process: 10 times the amount of water was used to extract twice, and each time was decocted for 1.5 h. The verification test showed that the contents of the five components were consistent with the predicted values, and the RSD values were all <5%. CONCLUSION The extraction process of heart-nourishing granule based on network pharmacology and Box-Behnken response surface methodology is stable and feasible, which provided an experimental basis for its further quality improvement.

ObjectiveTo assess the curative effects of Fangji Huangqi detumescence prescription （FHDP） on synovitis and polarization of synovial macrophages of knee osteoarthritis （KOA） model in rats induced by Hulth method. MethodThirty-six rats were randomly divided into sham operation group， model group， high-dose， medium-dose， and low-dose （29.16， 14.58， and 7.29 g·kg^-1） FHDP groups， and loxoprofen sodium （16.2 mg·kg^-1） group. KOA model in rats was induced by modified Hulth method. Six weeks after the operation， rats were given high， medium， and low concentrations of FHDP， normal saline （NS）， and loxoprofen sodium according to the group to intervene， and sacrificed after 2-week administration. Synovium and cartilage histopathological changes were observed after hematoxylin-eosin （HE） staining. Flow cytometry （FCM） and immunofluorescence （IF） test were used to evaluate the polarization of M1/M2 macrophages. Immunohistochemistry （IMC） and enzyme-linked immunosorbent assay （ELISA） were used to detect the related protein expression levels of macrophage polarization， such as interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， interleukin-10 （IL-10）， and matrix metalloproteinase-13 （MMP-13） in joint tissues and serum. ResultCompared with the sham operation group， Krenn and Mankin scores in the model group were significantly increased （P<0.01）. Compared with the model group， Krenn score was decreased in all administration groups （P<0.05， P<0.01）， but there was no significant difference in Mankin score in any administration groups. Compared with the sham operation group， M1/mø （CD38⁺） ratio in the model group was significantly increased （P<0.01）， and M2/mø （CD206⁺） ratio in the model group was decreased （P<0.05）. Compared with the model group， M1/mø ratio in the high， medium， and low-dose FHDP groups was decreased （P<0.05， P<0.01）， but M2/mø ratio was increased in all administration groups （the difference had no statistical significance）. Compared with the sham operation group， M1/M2 ratio in the model group was significantly increased （P<0.01）. Compared with the model group， M1/M2 ratio in all FHDP groups was significantly decreased （P<0.01）， and M1/M2 ratio in the high and medium-dose FHDP groups was lower than that in the loxoprofen sodium group （P<0.05）. Compared with the sham operation group， the levels of TNF-α， IL-1β， and MMP-13 in synovium and cartilage of the model group were significantly increased （P<0.01）， the level of IL-10 was significantly decreased （P<0.01）. Compared with the model group， the levels of TNF-α and IL-1β in synovium were decreased in all administration groups （P<0.05）， but the difference of the levels of MMP-13 and IL-10 in synovium had no statistical significance. The level of inflammatory mediators in cartilage was not affected in all administration groups. Compared with the sham operation group， the levels of TNF-α and IL-β in serum of the model group were significantly increased （P<0.01）， the level of IL-10 was decreased （P<0.05）. Compared with the model group， the level of TNF-α in the high-dose FHDP group was decreased （P<0.05）， and the level of IL-10 was increased in all administration groups （P<0.05， P<0.01）. The difference of the level of IL-β in all administration groups had no statistical significance. ConclusionFHDP attenuated the synovitis of KOA rats. FHDP exert the effect on the releasing of proinflammatory cytokines and MMP by inhibiting the polarization of M1 macrophages in synovium， and had no significant effect on the polarization of M2 macrophages. Modulating the imbalanced polarization of synovial macrophages was a possible mechanism of FHDP on attenuating synovitis and treating KOA.