Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition.

Objective To establish a rat model of Alzheimer's disease (AD) expressing human triple mutant amyloid precursor protein (APP) in the hippocampus, and to provide a model for the study of disease mechanisms and drug development. Methods Twenty-four 12-week-old SPF-grade female SD rats were randomly divided into a blank control group, a virus control group and an experimental group, with eight rats in each group; among them, the experimental group received a stereotaxic injection of adeno-associated virus (AAV) carrying the human triple mutant APP and NanoLuc luciferase genes into the hippocampus. In vivo imaging was used to observe viral expression in the brains of rats in each group, the novel object recognition test was used to assess the recognition memory of the rats in each group, real-time fluorescent quantitative PCR was used to detect the expression level of the APP gene, HE staining was used to examine the brain histopathology, Nissl staining was used to assess the hippocampal lesions, and immunohistochemistry was used to detect the deposition of amyloid β-protein (Aβ). Results In vivo imaging showed that reporter fluorescence was detected in the brains of rats in both experimental and virus control groups. Fluorescence quantitative PCR showed that the expression level of the APP gene was significantly increased in the brains of rats in the experimental group (P<0.01). Novel object recognition test revealed that the recognition memory of rats in the experimental group was significantly reduced compared with that of the blank control group (P<0.01). Six months after recombinant AAV virus infection, HE staining and Nissl staining of brain tissues showed that the number of neurons and Nissl bodies in the CA1 region of the hippocampus in the experimental group was reduced and disorganized; immuno-histochemistry testing of the CA1 region of the hippocampus and the pyramidal cell layer of the experimental group revealed prominent brown deposits, indicating Aβ protein deposition. Conclusion The rat model successfully established by stereotaxic injection and AAV-mediated delivery of human triple mutant APP gene exhibits typical AD features, providing a valuable animal model for studying AD pathology and developing drug therapies targeting Aβ protein deposition.

Objective:To explore the clinical efficacy of metoprolol combined with trimetazidine on elderly patients with coronary heart disease(CHD)and chronic heart failure(CHF).Methods:This randomized controlled trial enrolled 120 elderly CHD+CHF patients admitted to Army 73rd Group Military Hospital of Chinese PLA between June 2020 and June 2023.Patients were divided into control group(metoprolol based on routine treatment)and in-tervention group(additional trimetazidine therapy).Each group consisted of 60 patients,treated for 1 month.The clinical efficacy,left ventricular ejection fraction(LVEF),cardiac index(CI),left ventricular end-systolic diame-ter(LVESd),left ventricular end-diastolic diameter(LVEDd),serum levels of brain natriuretic peptide(BNP),high sensitive C-reactive protein(hsCRP),platelet a granule membrane protein-140(GMP-140),intercellular adhesion molecule-1(ICAM-1)and growth differentiation factor 15(GDF-15),and incidence of adverse reac-tions were compared between the two groups.Results:The total effective rate of the intervention group was significant-ly higher than that of the control group(95.00％vs.81.67％,P=0.023).Compared to patients in the control group,those in the intervention group had significant lower LVESd[(35.03±5.14)mm vs.(40.63±3.87)mm],LVEDd[(43.53±4.27)mm vs.(48.36±5.22)mm],levels of BNP[(94.35±7.55)pg/ml vs.(127.86±45.11)pg/ml],hsCRP[(0.91±0.28)mg/L vs.(1.47±0.52)mg/L],GMP-140[(7.14±1.06)μg/L vs.(9.37±1.59)μg/L],ICAM-1[(43.81±5.75)pg/ml vs.(52.74±5.83)pg/ml]andGDF-15[(891.46±62.51)pg/ml vs.(1025.57±110.08)pg/ml],and significant higher LVEF[(55.62±5.11)％vs.(47.35±8.61)％]and CI[(3.41±0.38)L·min-1·m-2 vs.(3.08±0.31)L·min-1·m-2](P＜0.001 all).There was no significant difference in the total inci-dence of adverse reactions between the intervention group and control group(8.33％vs.11.67％,P=0.543).Conclu-sion:Metoprolol combined trimetazidine may relieve myocardial inflammatory response and injury,and inhibit ventricular remodeling,thereby improve cardiac function in elderly patients with CHD and CHF.

Background and Aims:To overcome the limitations of the transoral endoscopic thyroidectomy vestibular approach,such as restricted operative space and high complication risks,our team proposed a modified technique—endoscopic thyroidectomy via oral vestibule and submandibular approach(ETOSA).Preliminary studies have confirmed its safety and feasibility.This study aims to systematically evaluate the key factors affecting postoperative complications and operative time in ETOSA,explore the interactions among these variables,and construct a learning curve model to support its broader clinical adoption.Methods:A retrospective analysis was conducted on 125 patients with papillary thyroid carcinoma who underwent ETOSA at Xiangya Hospital,Central South University,between March 2022 and March 2023.Clinical characteristics,surgical parameters,and postoperative complications were extracted.A random forest model was employed to identify the major influencing factors for complications and operative time,as well as their interaction effects.Partial dependence plots based on case sequence were used to generate the learning curve.Results:All 125 patients successfully underwent ETOSA with no conversion to open surgery.The median operative time was 95.0 min,and the median intraoperative blood loss was 15.0 mL.The overall postoperative complication rate was 16.0%,with no cases of permanent hypoparathyroidism or hypocalcemia.The average neck appearance score was 1.05,indicating high patient satisfaction.The random forest analysis identified case number,surgical extent,lymph node yield(LNY),Hashimoto's thyroiditis(HT),and body mass index(BMI)as the key predictors of postoperative complications,while surgical extent,case number,LNY,HT,and blood loss were the key factors affecting operative time.A significant positive interaction was observed between case number and both surgical extent and HT,particularly in the first 20 cases,suggesting a higher risk during the early learning phase.The learning curve analysis indicated that surgical proficiency stabilized after 20 cases.Operative time and complication rate in the proficient phase were significantly lower than those in the learning phase(90.0 min vs.102.5 min;11.4%vs.40.0%,both P<0.05).Conclusion:ETOSA is a safe and feasible technique characterized by minimal invasiveness,favorable cosmetic outcomes,and a relatively short learning curve.case number,surgical extent,LNY,HT,BMI,and blood loss are key factors affecting complications and operation time.

Objective To investigate the carrying status and drug resistance analysis of pathogenic bacteria in the gut of healthy population in Huadu District,Guangzhou,and provide reference for epidemiological re-search.Methods A total of 337 459 anal swabs were directly isolated and cultured from healthy individuals who underwent physical examinations from 2018 to 2023.600 anal swabs from 2022 were randomly selected for multi pathogen nucleic acid testing,direct isolation and culture,and enrichment culture.Identification of bacteria was conducted by using VITEK 2 Compact and VITEK MS,serological testing was used for determi-nation of type,and drug susceptibility testing of pathogenic bacteria was conducted.Results A total of 128 pathogenic bacteria were isolated from rectal swabs,including 52 strains of Salmonella,71 strains of Aero-monas,3 strains of Vibrio parahaemolyticus,and 2 strains of Shigella flexneri,with a total detection rate of 3.79/10 000.The detection rate of Salmonella in direct isolation and culture of 600 rectal swabs was 0.50％,the detection rate of enrichment culture was 1.00％,and the detection rate of multi-pathogen nucleic acid de-tection was 1.17％.52 strains of Salmonella were divided into 27 serotypes,and Salmonella Ⅰ 4,5,12:i:—was the dominant serotype.Aeromonas sobria was the dominant serotype in Aeromonas.The sensitivity of Salmonella to piperacillin/tazobactam,imipenem,cefepime,levofloxacin,and aztreonam was up to 92.00％or more,however,the phenomenon of multidrug resistance was severe,with a multidrug resistance rate as high as 40.38％.The resistance rate of Aeromonas to ampicillin and ampicillin/sulbactam was up to 85.00％or more,while Aeromonas was relatively sensitive to other antibiotics.Conclusion There are many species and types of intestinal pathogenic bacteria carried by healthy individuals in Huadu District,Guangzhou,with Aero-monas and Salmonella ranking first and second,and levofloxacin treatment could be preferred.The use of bac-terial culture and multi pathogen nucleic acid testing methods could improve the detection rate of intestinal pathogenic bacteria,and attention should be paid to the monitoring of intestinal pathogenic bacteria among rel-evant practitioners.

Objective To investigate the effect and mechanism of methyl oxofulnonone A(META)on high glucose(HG)-induced H9c2 cell injury.Methods H9c2 cells were divided into control group(normal culture),model group(55 mmol·L-1 glucose)and experimental-L,-M,-H groups(55 mmol·L-1 glucose+12.5,25.0,50.0 μmol·L-1 META).Cell viability was detected by cell counting kit-8;intracellular reactive oxygen species(ROS)level was detected by DCFH-DA fluorescent probe;intracellular adenosine triphosphate(ATP)content was detected by luciferase;and autophagy-related protein expression was detected by Western blotting.Results The optical density values of 72-hour cells in the control group,model group and experimental-M,-H groups were 0.91±0.03,0.61±0.01,0.69±0.02 and 0.72±0.03;the ROS levels were(40.75±1.53)％,(43.73±1.30)％,(30.87±1.27)％and(28.28±1.43)％;the ATP contents were(8.16±0.71),(4.03±0.29),(5.29±0.31)and(5.83±0.31)nmol·mg-1;the relative expression levels of autophagy-related gene 5 protein were 1.05±0.06,1.46±0.09,0.98±0.11 and 0.89±0.09;the relative expression levels of ubiquitin-binding protein were 1.05±0.10,0.55±0.13,0.89±0.04 and 0.98±0.04;the ratios of microtubule-associated protein 1 light chain 3 11/Ⅰ protein were 1.09±0.09,1.82±0.05,1.67±0.29 and 1.09±0.15,respectively.Among the above indicators,there were statistically significant differences between the model group and the control and experimental-M,-H groups(P＜0.05,P＜0.01).Conclusion META significantly ameliorates H9c2 cardiomyocyte damage caused by high glucose,ameliorates oxidative stress,protects mitochondrial respiration and inhibits autophagy.

Background and Aims:To overcome the limitations of the transoral endoscopic thyroidectomy vestibular approach,such as restricted operative space and high complication risks,our team proposed a modified technique—endoscopic thyroidectomy via oral vestibule and submandibular approach(ETOSA).Preliminary studies have confirmed its safety and feasibility.This study aims to systematically evaluate the key factors affecting postoperative complications and operative time in ETOSA,explore the interactions among these variables,and construct a learning curve model to support its broader clinical adoption.Methods:A retrospective analysis was conducted on 125 patients with papillary thyroid carcinoma who underwent ETOSA at Xiangya Hospital,Central South University,between March 2022 and March 2023.Clinical characteristics,surgical parameters,and postoperative complications were extracted.A random forest model was employed to identify the major influencing factors for complications and operative time,as well as their interaction effects.Partial dependence plots based on case sequence were used to generate the learning curve.Results:All 125 patients successfully underwent ETOSA with no conversion to open surgery.The median operative time was 95.0 min,and the median intraoperative blood loss was 15.0 mL.The overall postoperative complication rate was 16.0%,with no cases of permanent hypoparathyroidism or hypocalcemia.The average neck appearance score was 1.05,indicating high patient satisfaction.The random forest analysis identified case number,surgical extent,lymph node yield(LNY),Hashimoto's thyroiditis(HT),and body mass index(BMI)as the key predictors of postoperative complications,while surgical extent,case number,LNY,HT,and blood loss were the key factors affecting operative time.A significant positive interaction was observed between case number and both surgical extent and HT,particularly in the first 20 cases,suggesting a higher risk during the early learning phase.The learning curve analysis indicated that surgical proficiency stabilized after 20 cases.Operative time and complication rate in the proficient phase were significantly lower than those in the learning phase(90.0 min vs.102.5 min;11.4%vs.40.0%,both P<0.05).Conclusion:ETOSA is a safe and feasible technique characterized by minimal invasiveness,favorable cosmetic outcomes,and a relatively short learning curve.case number,surgical extent,LNY,HT,BMI,and blood loss are key factors affecting complications and operation time.

Objective To investigate the effect and mechanism of methyl oxofulnonone A(META)on high glucose(HG)-induced H9c2 cell injury.Methods H9c2 cells were divided into control group(normal culture),model group(55 mmol·L-1 glucose)and experimental-L,-M,-H groups(55 mmol·L-1 glucose+12.5,25.0,50.0 μmol·L-1 META).Cell viability was detected by cell counting kit-8;intracellular reactive oxygen species(ROS)level was detected by DCFH-DA fluorescent probe;intracellular adenosine triphosphate(ATP)content was detected by luciferase;and autophagy-related protein expression was detected by Western blotting.Results The optical density values of 72-hour cells in the control group,model group and experimental-M,-H groups were 0.91±0.03,0.61±0.01,0.69±0.02 and 0.72±0.03;the ROS levels were(40.75±1.53)％,(43.73±1.30)％,(30.87±1.27)％and(28.28±1.43)％;the ATP contents were(8.16±0.71),(4.03±0.29),(5.29±0.31)and(5.83±0.31)nmol·mg-1;the relative expression levels of autophagy-related gene 5 protein were 1.05±0.06,1.46±0.09,0.98±0.11 and 0.89±0.09;the relative expression levels of ubiquitin-binding protein were 1.05±0.10,0.55±0.13,0.89±0.04 and 0.98±0.04;the ratios of microtubule-associated protein 1 light chain 3 11/Ⅰ protein were 1.09±0.09,1.82±0.05,1.67±0.29 and 1.09±0.15,respectively.Among the above indicators,there were statistically significant differences between the model group and the control and experimental-M,-H groups(P＜0.05,P＜0.01).Conclusion META significantly ameliorates H9c2 cardiomyocyte damage caused by high glucose,ameliorates oxidative stress,protects mitochondrial respiration and inhibits autophagy.

Objective:To explore the clinical efficacy of metoprolol combined with trimetazidine on elderly patients with coronary heart disease(CHD)and chronic heart failure(CHF).Methods:This randomized controlled trial enrolled 120 elderly CHD+CHF patients admitted to Army 73rd Group Military Hospital of Chinese PLA between June 2020 and June 2023.Patients were divided into control group(metoprolol based on routine treatment)and in-tervention group(additional trimetazidine therapy).Each group consisted of 60 patients,treated for 1 month.The clinical efficacy,left ventricular ejection fraction(LVEF),cardiac index(CI),left ventricular end-systolic diame-ter(LVESd),left ventricular end-diastolic diameter(LVEDd),serum levels of brain natriuretic peptide(BNP),high sensitive C-reactive protein(hsCRP),platelet a granule membrane protein-140(GMP-140),intercellular adhesion molecule-1(ICAM-1)and growth differentiation factor 15(GDF-15),and incidence of adverse reac-tions were compared between the two groups.Results:The total effective rate of the intervention group was significant-ly higher than that of the control group(95.00％vs.81.67％,P=0.023).Compared to patients in the control group,those in the intervention group had significant lower LVESd[(35.03±5.14)mm vs.(40.63±3.87)mm],LVEDd[(43.53±4.27)mm vs.(48.36±5.22)mm],levels of BNP[(94.35±7.55)pg/ml vs.(127.86±45.11)pg/ml],hsCRP[(0.91±0.28)mg/L vs.(1.47±0.52)mg/L],GMP-140[(7.14±1.06)μg/L vs.(9.37±1.59)μg/L],ICAM-1[(43.81±5.75)pg/ml vs.(52.74±5.83)pg/ml]andGDF-15[(891.46±62.51)pg/ml vs.(1025.57±110.08)pg/ml],and significant higher LVEF[(55.62±5.11)％vs.(47.35±8.61)％]and CI[(3.41±0.38)L·min-1·m-2 vs.(3.08±0.31)L·min-1·m-2](P＜0.001 all).There was no significant difference in the total inci-dence of adverse reactions between the intervention group and control group(8.33％vs.11.67％,P=0.543).Conclu-sion:Metoprolol combined trimetazidine may relieve myocardial inflammatory response and injury,and inhibit ventricular remodeling,thereby improve cardiac function in elderly patients with CHD and CHF.

OBJECTIVE: Myocardial ischemia/reperfusion injury (MIRI) is an obstacle to the success of cardiac reperfusion therapy. This study explores whether luteolin can mitigate MIRI by regulating the p53 signaling pathway. METHODS: Model mice were subjected to a temporary surgical ligation of the left anterior descending coronary artery, and administered luteolin. The myocardial infarct size, myocardial enzyme levels, and cardiac function were measured. Latent targets and signaling pathways were screened using network pharmacology and molecular docking. Then, proteins related to the p53 signaling pathway, apoptosis and oxidative stress were measured. Hypoxia/reoxygenation (HR)-incubated HL1 cells were used to validate the effects of luteolin in vitro. In addition, a p53 agonist and an inhibitor were used to investigate the mechanism. RESULTS: Luteolin reduced the myocardial infarcted size and myocardial enzymes, and restored cardiac function in MIRI mice. Network pharmacology identified p53 as a hub target. The bioinformatic analyses showed that luteolin had anti-apoptotic and anti-oxidative properties. Additionally, luteolin halted the activation of p53, and prevented both apoptosis and oxidative stress in myocardial tissue in vivo. Furthermore, luteolin inhibited cell apoptosis, JC-1 monomer formation, and reactive oxygen species elevation in HR-incubated HL1 cells in vitro. Finally, the p53 agonist NSC319726 downregulated the protective attributes of luteolin in the MIRI mouse model, and both luteolin and the p53 inhibitor pifithrin-α demonstrated a similar therapeutic effect in the MIRI mice. CONCLUSION Luteolin effectively treats MIRI and may ameliorate myocardial damage by regulating apoptosis and oxidative stress through its targeting of the p53 signaling pathway. Please cite this article as: Zhai P, Ouyang XH, Yang ML, Lin L, Li JY, Li YM, Cheng X, Zhu R, Hu DS. Luteolin protects against myocardial ischemia/reperfusion injury by reducing oxidative stress and apoptosis through the p53 pathway. J Integr Med. 2024; 22(6): 652-664.
Luteolin/pharmacology* ; Animals ; Myocardial Reperfusion Injury/metabolism* ; Oxidative Stress/drug effects* ; Tumor Suppressor Protein p53/genetics* ; Apoptosis/drug effects* ; Mice ; Signal Transduction/drug effects* ; Male ; Disease Models, Animal ; Mice, Inbred C57BL ; Myocardial Infarction/prevention & control* ; Reactive Oxygen Species/metabolism*