ObjectiveThis paper aims to observe the syndrome improvement and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome）. MethodsThrough a multi-center randomized controlled methodology design，confirmed influenza cases were collected from October 2022 to April 2023 in the pediatrics department of eight hospitals，such as Dongfang Hospital of Beijing University of Chinese Medicine. A total of 180 children with influenza and heat accumulation in the lung and stomach syndrome conforming to the standard were recruited through the clinic. The sick children meeting the inclusion criteria were randomly divided into groups by a block-randomized method. The children in the experimental group were treated with Qingxuan Daozhi formula for five days，and those in the control group were treated with Oseltamivir Phosphate Granules for five days. The primary efficacy indicator was the negative conversion rate of influenza antigen detection. Secondary efficacy indicators were the Canadian acute respiratory illness and flu scale （CARIFS） and the incidence of complications，severe cases， and critical cases. Follow-up observation was conducted on the day of enrollment，48 hours after medication，72 hours after medication， and （6+1） d after medication. ResultsOne hundred and eighty participants were randomly assigned to the experimental group （90 cases） or the control group （90 cases）. All participants were followed up during the study. Comparison of influenza antigen detection results in the primary efficacy indicators showed that the average time of negative influenza antigen conversion in the experimental group was （5.29±1.25） d，and that in the control group was （5.40±1.68） d，without a statistically significant difference. After five days of intervention，52 cases in the experimental group and 51 cases in the control group converted to negative，without a statistically significant difference. CARIFS score results in the secondary efficacy indicators showed that during 72 hours after intervention，there were statistically significant differences between the experimental group and the control group in three dimensions， including headache，muscle soreness， and the need for extra care （P<0.05）. On the （6+1） days after the intervention，the differences in both the experimental group and the control group were statistically significant in 10 dimensions， including sore throat，bad sleep，uncomfortable feeling，poor spirit and fatigue，crying more than usual，the need for extra care，symptom，function，influence on parents，and total score （P<0.05）. The comparison results within the group in the dimensional scores of symptom， function， and influence on parents，as well as the CARIFS total score showed that with the delay of follow-up time，scores of both groups decreased significantly，with a statistically significant difference （P<0.01）. Inter-group comparison results showed that the mean score of the experimental group was higher than that of the control group at the time of enrollment. With the progress of intervention，the score of the experimental group was significantly decreased compared with that of the control group. At the end of follow-up，the mean score of the experimental group was lower than that of the control group，with no statistically significant difference. In terms of the incidence of complications，severe cases， and critical cases， there were no complications，severe cases， and critical cases in the two groups，without a statistically significant difference. ConclusionThe symptom improvement effect and negative antigen conversion rate of Qingxuan Daozhi formula in the treatment of influenza in children （heat accumulation in the lung and stomach syndrome） are not inferior to Oseltamivir Phosphate granules， and children's acceptance is better. It can be more widely used in clinical treatment of influenza in children （heat accumulation in the lung and stomach syndrome）.

ObjectiveTo investigate the effect and mechanism of Shengxian Yixin Granule （升陷益心颗粒， SYG） in treating heart failure with preserved ejection fraction （HFpEF）. MethodsExperiment 1： Forty male C57BL/6 mice were randomly divided into control group， model group， low-dose ［25 mg/（kg·d）］， medium-dose ［50 mg/（kg·d）］， and high-dose ［（75 mg/（kg·d）］ SYG groups， with 8 mice per group. Except for the control group， HFpEF was induced by a high-fat diet combined with Nω-nitro-L-arginine methyl ester in drinking water for 16 weeks. After 10 weeks of modeling， treatment groups received corresponding doses of SYG by gavage， while the control group and model group received equal volume of normal saline， for 6 weeks. After 6 weeks of administration， the body weight， blood pressure including systolic blood pressure （SBP） and diastolic blood pressure （DBP）， and heart weight-to-tibia length ratio （HW/TL） were recorded. Echocardiography was used to evaluate left ventricular ejection fraction （LVEF） and the ratio of early to late diastolic transmitral flow velocity （E/A）. Plasma N-terminal pro-brain natriuretic peptide （NT-proBNP） and C-reactive protein （CRP） levels were measured by ELISA. The expression level of brain natriuretic peptide （BNP） in heart tissue was detected by qRT-PCR. Blood lipid levels， including low-density lipoprotein cholesterol （LDL-C）， triglycerides （TG）， and total cholesterol （TC）， were determined by a biochemical analyzer. Splenic proportions of M1 macrophages， M2 macrophages， T helper 17 （Th17） cells， and regulatory T （Treg） cells were analyzed by flow cytometry. Experiment 2： RAW 264.7 cells were stimulated with lipopolysaccharide （LPS） to establish an M1 macrophage polarization model， and then treated with SYG-containing serum for 24 h. The cells were divided into four groups， including blank serum group， high-dose SYG-containing serum group， LPS group， and LPS plus high-dose SYG-containing serum group. Combined with asignal transducer and activator of transcription 3 （Stat3） inhibitor for 24 h， the cells were divided into a lipopolysaccharide group， a high-dose SYG-containing serum group， an inhibitor group， and high-dose SYG-containing serum plus inhibitor group. The expression levels of inducible nitric oxide synthase （iNOS） and interleukin-1β （IL-1β） in each group were detected by qRT-PCR. The protein levels of iNOS， IL-1β， Stat3， and phosphorylated Stat3 （p-Stat3） were detected by Western Blotting. ResultsExperiment 1： Compared to the control group， the model group showed significantly decreased E/A ratio， and increased cardiac BNP expression， plasma NT-proBNP，HW/TL ratio， body weight， TC， LDL-C， SBP， and DBP， as well as shortened exhaustion distance and exhaustion time （P＜0.05 or P＜0.01）. Compared to the model group， the E/A ratio increased in the low-， medium-， and high-dose SYG groups； cardiac BNP expression decreased in the low- and high-dose groups， and plasma NT-proBNP levels decreased in the high-dose group； the HW/TL ratio and body weight decreased， while exhaustion distance and exhaustion time increased in the medium- and high-dose groups； TG levels decreased in the low- and medium-dose groups， and LDL-C levels decreased in the medium- and high-dose groups； SBP and DBP decreased in the low-， medium-， and high-dose groups （P＜0.05 or P＜0.01）. Compared to the low-dose SYG group， body weight decreased， and both exhaustion distance and exhaustion time increased in the medium- and high-dose groups （P＜0.05 or P＜0.01）. Compared to the medium-dose SYG group， the exhaustion time increased in the high-dose SYG group （P＜0.01）. Experiment 2： In the LPS‑induced M1 macrophage polarization model， compared to the blank serum group， the LPS group showed significantly increased mRNA and protein expression of iNOS and IL‑1β， as well as the p‑Stat3/Stat3 ratio （P＜0.01）. Compared to the LPS group， the high‑dose SYG-containing serum plus LPS group showed significantly decreased mRNA and protein expression of iNOS and IL‑1β， as well as the p‑Stat3/Stat3 ratio （P＜0.05 or P＜0.01）. Compared to the LPS group， the inhibitor group also showed significantly reduced these parameters （P＜0.05 or P＜0.01）. ConclusionSYG can improve cardiac function， ventricular remodeling， exercise capacity， and lipid metabolism in HFpEF mice， while inhibiting inflammatory responses. Its mechanism may be related to the regulation of the Stat3 signaling pathway and inhibition of M1 macrophage polarization.

ObjectiveTo investigate the effect and mechanism of Shengxian Yixin Granule （升陷益心颗粒， SYG） in treating heart failure with preserved ejection fraction （HFpEF）. MethodsExperiment 1： Forty male C57BL/6 mice were randomly divided into control group， model group， low-dose ［25 mg/（kg·d）］， medium-dose ［50 mg/（kg·d）］， and high-dose ［（75 mg/（kg·d）］ SYG groups， with 8 mice per group. Except for the control group， HFpEF was induced by a high-fat diet combined with Nω-nitro-L-arginine methyl ester in drinking water for 16 weeks. After 10 weeks of modeling， treatment groups received corresponding doses of SYG by gavage， while the control group and model group received equal volume of normal saline， for 6 weeks. After 6 weeks of administration， the body weight， blood pressure including systolic blood pressure （SBP） and diastolic blood pressure （DBP）， and heart weight-to-tibia length ratio （HW/TL） were recorded. Echocardiography was used to evaluate left ventricular ejection fraction （LVEF） and the ratio of early to late diastolic transmitral flow velocity （E/A）. Plasma N-terminal pro-brain natriuretic peptide （NT-proBNP） and C-reactive protein （CRP） levels were measured by ELISA. The expression level of brain natriuretic peptide （BNP） in heart tissue was detected by qRT-PCR. Blood lipid levels， including low-density lipoprotein cholesterol （LDL-C）， triglycerides （TG）， and total cholesterol （TC）， were determined by a biochemical analyzer. Splenic proportions of M1 macrophages， M2 macrophages， T helper 17 （Th17） cells， and regulatory T （Treg） cells were analyzed by flow cytometry. Experiment 2： RAW 264.7 cells were stimulated with lipopolysaccharide （LPS） to establish an M1 macrophage polarization model， and then treated with SYG-containing serum for 24 h. The cells were divided into four groups， including blank serum group， high-dose SYG-containing serum group， LPS group， and LPS plus high-dose SYG-containing serum group. Combined with asignal transducer and activator of transcription 3 （Stat3） inhibitor for 24 h， the cells were divided into a lipopolysaccharide group， a high-dose SYG-containing serum group， an inhibitor group， and high-dose SYG-containing serum plus inhibitor group. The expression levels of inducible nitric oxide synthase （iNOS） and interleukin-1β （IL-1β） in each group were detected by qRT-PCR. The protein levels of iNOS， IL-1β， Stat3， and phosphorylated Stat3 （p-Stat3） were detected by Western Blotting. ResultsExperiment 1： Compared to the control group， the model group showed significantly decreased E/A ratio， and increased cardiac BNP expression， plasma NT-proBNP，HW/TL ratio， body weight， TC， LDL-C， SBP， and DBP， as well as shortened exhaustion distance and exhaustion time （P＜0.05 or P＜0.01）. Compared to the model group， the E/A ratio increased in the low-， medium-， and high-dose SYG groups； cardiac BNP expression decreased in the low- and high-dose groups， and plasma NT-proBNP levels decreased in the high-dose group； the HW/TL ratio and body weight decreased， while exhaustion distance and exhaustion time increased in the medium- and high-dose groups； TG levels decreased in the low- and medium-dose groups， and LDL-C levels decreased in the medium- and high-dose groups； SBP and DBP decreased in the low-， medium-， and high-dose groups （P＜0.05 or P＜0.01）. Compared to the low-dose SYG group， body weight decreased， and both exhaustion distance and exhaustion time increased in the medium- and high-dose groups （P＜0.05 or P＜0.01）. Compared to the medium-dose SYG group， the exhaustion time increased in the high-dose SYG group （P＜0.01）. Experiment 2： In the LPS‑induced M1 macrophage polarization model， compared to the blank serum group， the LPS group showed significantly increased mRNA and protein expression of iNOS and IL‑1β， as well as the p‑Stat3/Stat3 ratio （P＜0.01）. Compared to the LPS group， the high‑dose SYG-containing serum plus LPS group showed significantly decreased mRNA and protein expression of iNOS and IL‑1β， as well as the p‑Stat3/Stat3 ratio （P＜0.05 or P＜0.01）. Compared to the LPS group， the inhibitor group also showed significantly reduced these parameters （P＜0.05 or P＜0.01）. ConclusionSYG can improve cardiac function， ventricular remodeling， exercise capacity， and lipid metabolism in HFpEF mice， while inhibiting inflammatory responses. Its mechanism may be related to the regulation of the Stat3 signaling pathway and inhibition of M1 macrophage polarization.

Objective The aim of the study is to analyze the incidence patterns and epidemiological characteristics of parasitic and vector-borne diseases in the Minhang and Jinshan districts of Shanghai, and provide evidence-based information for optimizing prevention and control measures. Methods We collected information from epidemic reports and epidemiological investigation forms for cases of parasitic and vector-borne diseases from 2005 to 2024 in Shanghai's Minhang and Jinshan districts. Descriptive epidemiological method were adopted to conduct a comparative analysis of incidence trends, epidemic characteristics, and infection sources. Results A total of 229 cases in 7 categories were reported between 2005 and 2024, with an average annual incidence rate of 0.36/100 000. The average annual incidence rate in Minhang District was significantly higher than that in Jinshan District. The most prevalent diseases were malaria and dengue fever, followed by Japanese encephalitis. Monthly case reports were available, predominantly from June to October, with a peak in July-August. A statistically significant difference was observed between the two districts in case distribution during June-October (P＜0.001), whereas no significant difference was found for July-August (P=0.331). The majority of cases were young men (111/229,48.47%), aged 20-39 years. Migrant workers/farmers constituted the largest occupational group (55/229, 24.02%). Registered residents of other provinces and cities (147/229,64.19%), and the main source of infection was overseas importation (112/229,48.91%). There were no statistically significant differences in gender and household-registration composition between the two districts (P＞0.05). Significant differences existed in age, occupation, and source of infection (P＜0.05). The imported diseases were malaria and dengue fever, with Africa and Southeast Asia being the primary source locations. Malaria was the predominant disease imported from other provinces and cities, with cases having been imported from abroad since 2012. Local infections were mainly Japanese encephalitis, followed by malaria (with the last local case reported in 2010). Cases between 2005 and 2009 were predominantly imported from other provinces and cities. The proportion of cases imported from abroad increased sharply after 2010, particularly in Minhang District. Local infections sometimes arise in Jinshan District, with Japanese encephalitis serving as the primary type. Conclusions Incidence rates of parasitic and vector-borne diseases in Minhang and Jinshan districts have generally remained low. To further reduce this disease burden, it will be necessary to strengthen joint measures for prevention and control, eliminate the environmental risk factors of vector organisms, enhance monitoring and precise control of the environment and inbound personnel to prevent the occurrence and transmission of parasitic and vector-borne diseases.

Objective To introduce an innovative technique, the "balance-shaped sternal elevation device" and its application in the subxiphoid uniportal video-assisted thoracoscopic surgery (VATS) for anterior mediastinal masses resection. Methods Patients who underwent single-port thoracoscopic assisted anterior mediastinal tumor resection through the xiphoid process at the Department of Thoracic Surgery, West China Hospital, Sichuan University from May to June 2024 were included, and their clinical data were analyzed. Results A total of 7 patients were included, with 3 males and 4 females, aged 28-72 years. The diameter of the tumor was 1.9-17.0 cm. The operation time was 62-308 min, intraoperative blood loss was 5-100 mL, postoperative chest drainage tube retention time was 0-9 days, pain score on the 7th day after surgery was 0-2 points, and postoperative hospital stay was 3-12 days. All patients underwent successful and complete resection of the masses and thymus, with favorable postoperative recovery. Conclusion The "balance-shaped sternal elevation device" effectively expands the retrosternal space, providing surgeons with satisfactory surgical views and operating space. This technique significantly enhances the efficacy and safety of minimally invasive surgery for anterior mediastinal masses, reduces trauma and postoperative pain, and accelerates patient recovery, demonstrating important clinical significance and application value.

ObjectiveTo evaluate the efficacy and safety of Xiaoji Hufei Formula in protecting children with close contact exposure to influenza， and to provide reference and evidence-based support for better clinical prevention and treatment of influenza in children. MethodsA multicenter， prospective， non-randomized， parallel， controlled trial was conducted from October 2021 to May 2022 in five hospitals， including Dongfang Hospital of Beijing University of Chinese Medicine. Confirmed influenza cases and influenza-like illness （ILI） cases were collected， and eligible children with close contact exposure to these cases were recruited in the outpatient clinics. According to whether the enrolled close contacts were willing to take Xiaoji Hufei formula for influenza prevention， they were assigned to the observation group （108 cases） or the control group （108 cases）. Follow-up visits were conducted on days 7 and 14 after enrollment. The primary outcomes were the incidence of ILI and the rate of laboratory-confirmed influenza. Secondary outcomes included traditional Chinese medicine （TCM） symptom score scale for influenza， influenza-related emergency （outpatient） visit rate， influenza hospitalization rate， and time to onset after exposure to influenza cases. ResultsA total of 216 participants were enrolled， with 108 in the observation group and 108 in the control group. Primary outcomes： （1） Incidence of ILI： The incidence was 12.0% （13/108） in the observation group and 23.1% （25/108） in the control group， with the observation group showing a significantly lower incidence （χ²=4.6， P<0.05）. （2） Influenza confirmation rate： 3.7% （4/108） in the observation group and 4.6% （5/108） in the control group， with no statistically significant difference. Secondary outcomes： （1） TCM symptom score scale： after onset， nasal congestion and runny nose scores differed significantly between the two groups （P<0.05）， while other symptoms such as fever， sore throat， and cough showed no significant differences. （2） Influenza-related emergency （outpatient） visit rate： 84.6% （11 cases） in the observation group and 96.0% （24 cases） in the control group， with no significant difference. （3） Time to onset after exposure： The median onset time after exposure to index patients was 7 days in the observation group and 4 days in the control group， with a statistically significant difference （P<0.05）. ConclusionIn previously healthy children exposed to infectious influenza cases under unprotected conditions， Xiaoji Hufei formula prophylaxis significantly reduced the incidence of ILI. Xiaoji Hufei Formula can be recommended as a specific preventive prescription for influenza in children.

ObjectiveTo evaluate the efficacy and safety of Xiaoji Hufei Formula in protecting children with close contact exposure to influenza， and to provide reference and evidence-based support for better clinical prevention and treatment of influenza in children. MethodsA multicenter， prospective， non-randomized， parallel， controlled trial was conducted from October 2021 to May 2022 in five hospitals， including Dongfang Hospital of Beijing University of Chinese Medicine. Confirmed influenza cases and influenza-like illness （ILI） cases were collected， and eligible children with close contact exposure to these cases were recruited in the outpatient clinics. According to whether the enrolled close contacts were willing to take Xiaoji Hufei formula for influenza prevention， they were assigned to the observation group （108 cases） or the control group （108 cases）. Follow-up visits were conducted on days 7 and 14 after enrollment. The primary outcomes were the incidence of ILI and the rate of laboratory-confirmed influenza. Secondary outcomes included traditional Chinese medicine （TCM） symptom score scale for influenza， influenza-related emergency （outpatient） visit rate， influenza hospitalization rate， and time to onset after exposure to influenza cases. ResultsA total of 216 participants were enrolled， with 108 in the observation group and 108 in the control group. Primary outcomes： （1） Incidence of ILI： The incidence was 12.0% （13/108） in the observation group and 23.1% （25/108） in the control group， with the observation group showing a significantly lower incidence （χ²=4.6， P<0.05）. （2） Influenza confirmation rate： 3.7% （4/108） in the observation group and 4.6% （5/108） in the control group， with no statistically significant difference. Secondary outcomes： （1） TCM symptom score scale： after onset， nasal congestion and runny nose scores differed significantly between the two groups （P<0.05）， while other symptoms such as fever， sore throat， and cough showed no significant differences. （2） Influenza-related emergency （outpatient） visit rate： 84.6% （11 cases） in the observation group and 96.0% （24 cases） in the control group， with no significant difference. （3） Time to onset after exposure： The median onset time after exposure to index patients was 7 days in the observation group and 4 days in the control group， with a statistically significant difference （P<0.05）. ConclusionIn previously healthy children exposed to infectious influenza cases under unprotected conditions， Xiaoji Hufei formula prophylaxis significantly reduced the incidence of ILI. Xiaoji Hufei Formula can be recommended as a specific preventive prescription for influenza in children.

BACKGROUND: Chronic kidney disease (CKD)-associated anemia (CKD-anemia) is associated with poor survival, and hemoglobin targets are often not achieved with current therapies. Phase 3 trials have demonstrated the treatment efficacy of roxadustat for CKD-anemia. This phase 4 study aims to evaluate the long-term (52-week) safety and effectiveness of roxadustat in a broad real-world patient population with CKD-anemia with and without dialysis in China. METHODS: This Phase 4 multicenter, open-label, prospective study, conducted from 24 November 2020 to 11 November 2022, evaluated the long-term safety and effectiveness of roxadustat for CKD-anemia in China. Patients aged ≥18 years with CKD-anemia with or without dialysis were included. The initial oral dose was 70-120 mg (weight-based followed by dose adjustment) over 52 weeks. The primary endpoint was safety based on adverse events (AEs). The secondary endpoints were hemoglobin changes from baseline and the proportion of patients who achieved mean hemoglobin ≥100 g/L. Effectiveness evaluable populations 1 (EE1) and EE2 included roxadustat-naïve and previously roxadustat-treated patients, respectively. The safety analysis set (SAF) included all patients who received ≥1 occasion. RESULTS: The EE1, EE2, and SAF populations included 1804, 193, and 2021 patients, respectively. In the SAF, the mean age was 50 ± 14 years, and 1087 patients (53.8%) were male. Mean baseline hemoglobin was 96.9 ± 14.0 g/L in EE1 and 100.3 ± 12.9 g/L in EE2. In EE1, the mean (95% confidence interval) hemoglobin changes from baseline over weeks 24-36 and 36-52 were 14.2 (13.5-14.9) g/L and 14.3 (13.5-15.0) g/L, respectively. Over weeks 24-36 and 36-52, 83.3% and 86.1% of patients in EE1 and 82.7% and 84.7% in EE2 achieved mean hemoglobin ≥100 g/L, respectively. In the SAF, 1643 (81.3%) patients experienced treatment-emergent AEs (TEAEs). Overall, 219 (10.8%) patients experienced drug-related TEAEs. Thirty-eight (1.9%) patients died of TEAEs (unrelated to the study drug). Vascular access thrombosis was uncommon. CONCLUSIONS: Roxadustat (52 weeks) increased hemoglobin and maintained the treatment target in Chinese patients with CKD-anemia with acceptable safety, supporting its use in real-world settings. REGISTRATION Chinese Clinical Trial Registry ( www.chictr.org.cn ) ChiCTR2100046322; CDE ( www.chinadrugtrials.org.cn ) CTR20201568.
Humans ; Male ; Female ; Anemia/etiology* ; Middle Aged ; Renal Insufficiency, Chronic/complications* ; Glycine/adverse effects* ; Isoquinolines/adverse effects* ; Aged ; Prospective Studies ; Adult ; Hemoglobins/metabolism* ; Treatment Outcome ; China ; Registries ; East Asian People

Skeletal muscle atrophy is characterized by a reduction in both the size and quantity of skeletal muscle fibers, resulting in impaired muscle strength and function. It mainly includes disuse muscle atrophy, aging muscle atrophy, denervated muscle atrophy and muscle atrophy caused by disease etc. As a cost-effective way, exercise has been widely used in the prevention and treatment of skeletal muscle atrophy, but its mechanism for improving skeletal muscle atrophy remains unclear. Recent studies have indicated that insulin-like growth factor 1 (IGF-1) plays an important role in improving muscle atrophy through exercise, in addition to promoting the survival of neurons, lowering blood sugar, and anti-inflammation. This article reviews recent findings on the mechanisms by which IGF-1 mediates exercise-induced improvement in skeletal muscle atrophy, providing a theoretical basis for the prevention and treatment of this disease.
Insulin-Like Growth Factor I/physiology* ; Muscular Atrophy/therapy* ; Humans ; Exercise/physiology* ; Muscle, Skeletal ; Animals ; Insulin-Like Peptides

This study aimed to investigate the underlying mechanisms of Duhuo Jisheng Decoction(DJD) in the prevention and treatment of knee osteoarthritis(KOA). Forty SPF New Zealand rabbits were randomly divided using SPSS 26.0 software into five groups: blank group, model group, low-dose DJD group, high-dose DJD group, and high-dose DJD+phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt)/mammalian target of rapamycin(mTOR) signaling pathway activator group(high-dose DJD+740Y-P group), with eight rabbits in each group. Except for the blank group, the KOA model was established in the other groups using papain injection into the knee joint cavity combined with forced flexion of the knee joint. The day after modeling, the blank group and model group were given normal saline at 10 mL·kg~(-1) by gavage, the low-dose DJD group received DJD at 8.8 g·kg~(-1) by gavage, the high-dose DJD group received DJD at 35.2 g·kg~(-1) by gavage, and the high-dose DJD+740Y-P group received DJD at 35.2 g·kg~(-1) by gavage along with 740Y-P at 0.15 μmoL·kg~(-1) injected via the auricular vein. All groups received treatment continuously for four weeks. After modeling and intervention, behavioral observations were performed for all groups, and after the intervention, imaging assessments of the knee joints were conducted. Cartilage from the knee joints was collected, and gross morphological changes were observed. Pathological changes in cartilage tissue were examined using hematoxylin-eosin(HE) staining. The results of these observations were quantitatively evaluated using the Lequesne MG score, Kellgren-Lawrence(K-L) grading, Pelletier score, and Mankin score. ELISA was used to measure the levels of interleukin-1β(IL-1β), interleukin-18(IL-18), and matrix metalloproteinase 13(MMP13) in cartilage tissue. Real-time RT-PCR was used to detect the mRNA expression levels of PI3K, Akt, mTOR, Nod-like receptor protein 3(NLRP3), cysteine protease 1(caspase-1), and gasdermin D(GSDMD) in cartilage tissue. Western blot was employed to measure the protein expression levels of PI3K, Akt, mTOR, NLRP3, caspase-1, and GSDMD. The results showed that compared with the blank group, the model group exhibited significant knee joint degeneration, increased Lequesne MG score, K-L grading, Pelletier score, and Mankin score, elevated levels of IL-1β, IL-18, and MMP13 in cartilage tissue, activation of PI3K, Akt, and mTOR phosphorylation along with increased mRNA expression levels, and elevated protein and mRNA expression levels of NLRP3, caspase-1, and GSDMD. Compared with the model group, these indicators were reversed in both the low-dose and high-dose DJD groups, with the high-dose group showing greater decline degree than the low-dose DJD group. However, compared with the high-dose DJD group, the improvements in knee joint degeneration were less pronounced in the high-dose DJD+740Y-P group, with increased Lequesne MG score, K-L grading, Pelletier score, Mankin score, elevated levels of IL-1β, IL-18, and MMP13, activation of PI3K, Akt, and mTOR phosphorylation along with increased mRNA expression, and increased protein and mRNA expression levels of NLRP3, caspase-1, and GSDMD. In conclusion, DJD is effective and safe in the treatment of KOA, and its mechanism may be related to the inhibition of PI3K/Akt/mTOR signaling pathway-mediated pyroptosis in cartilage tissue, thereby improving knee joint bone structure, reducing the inflammatory response, and preventing cartilage matrix degradation.
Animals ; Drugs, Chinese Herbal/administration & dosage* ; Rabbits ; TOR Serine-Threonine Kinases/genetics* ; Osteoarthritis, Knee/genetics* ; Proto-Oncogene Proteins c-akt/genetics* ; Signal Transduction/drug effects* ; Male ; Disease Models, Animal ; Pyroptosis/drug effects* ; Phosphatidylinositol 3-Kinases/genetics* ; Humans ; Female