ObjectiveZheng’s San Qi San (ZSQS) power, a classic traditional Chinese medicine (TCM) formula, is used for treating soft tissue injuries involving muscles, tendons, and ligaments. However, its underlying therapeutic mechanisms remain unclear. This study aimed to screen and identify pharmaceutically active ingredients and their candidate biomolecule targets, and further elucidate the molecular mechanism of ZSQS in the treatment of skeletal muscle injury. MethodsNetwork pharmacology was employed to construct “ZSQS-component-target”, “protein-protein interaction (PPI)” and “active ingredient-core protein-pathway” networks to predict the key active ingredients and potential core targets of ZSQS for skeletal muscle injury. The predicted results were then validated via microarray data from the GEO database. Molecular docking was then performed to assess the binding ability between the screened active ingredients of ZSQS and the candidate core targets. Moreover, liquid chromatography-mass spectrometry (LC-MS) was used for qualitative and quantitative analysis to verify the active components of the drug and ZSQS serum. Finally, an animal model of eccentric exercise-induced skeletal muscle injury and a myotube cell model of oxidative stress-induced injury were established to validate the effects of ZSQS and its interventional effects on the biological functions of critical targets, thereby demonstrating the potential therapeutic mechanism of ZSQS. ResultsAmong the 111 active components identified in ZSQS and their corresponding 204 targets related to the skeletal muscle injury repair process, 14 core targets (including AKT1) and 4 core active components (quercetin, luteolin, kaempferol, and β‑sitosterol) were screened out, while the corresponding metabolites of quercetin, luteolin and kaempferol were detected in the ZSQS serum. Among these targets, 5 candidate genes (IL-6, CASP3, HIF1A, STAT3, and JUN) overlapped with the differential expression screening results with GEO data, and IL-6 was confirmed to be enriched in the PI3K/AKT pathway. Combined with the prediction results of the AKT expression levels, these findings suggest that the phosphorylation level of AKT1 plays a core role in the therapeutic mechanism of ZSQS. Molecular docking analysis further revealed that the PH domain of AKT1 had high binding energy with all 4 core active components, as verified by LC-MS. Finally, animal model studies have shown the promoting effect of ZSQS administration on skeletal muscle injury repair and its possible antioxidant damage mechanism. Cell model studies further demonstrated that ZSQS-containing serum, core active ingredient combination therapy, and quercetin monomer could increase the phosphorylation level of AKT, promote the nuclear translocation of Nrf2, upregulate the expression of downstream antioxidant enzymes (SOD, GPx, and GR), and inhibit the expression of inflammatory factors (IL-6 and TNF-α), thereby alleviating oxidative stress and the inflammatory response. ConclusionZSQS alleviates skeletal muscle injury mainly by activating the AKT/Nrf2 signaling pathway, enhancing cellular antioxidant and anti-inflammatory capabilities. The results of this study provide a scientific basis for the clinical application and modernized development of ZSQS.

BACKGROUND:The knee osteoarthritis,a major cause of disability in the elderly,is currently treated with surgical procedures including unicompartmental knee arthroplasty in end-stage patients,but the recovery of muscle strength in patients after unicompartmental knee arthroplasty is not well understood.OBJECTIVE:To analyze the short-term and medium-term knee isometric muscle strength characteristics of patients with unilateral knee osteoarthritis after unicompartmental knee arthroplasty,to compare with those of the unaffected side,and to analyze the recovery of muscle strength in the affected limb.METHODS:Patients undergoing unicompartmental knee arthroplasty in the Qingdao Municipal Hospital from June to December 2023 were recruited to undergo standardized functional rehabilitation exercises.The A8-2M multi-joint isokinetic training and testing system was used to detect the peak torque and the maximum one-repetition work of the extensor muscle of the unaffected side and the affected side of the knee joint at the angular velocity of 60(°)/s and the range of joint motion of 0°-90° before,1 and 3 months after surgery.RESULTS AND CONCLUSION:(1)At an angular velocity of 60(°)/s,the peak moment and maximum one-repetition work of the flexor and extensor muscles of the affected side were lower than those of the unaffected side preoperatively(P＜0.05).(2)The peak moment and maximum one-repetition work of the flexor and extensor muscles were significantly lower than those of the unaffected side 1 month postoperatively(P＜0.05).Meanwhile,the peak moment of the flexor and extensor muscles and the maximum one-repetition work of the flexor and extensor muscles on the affected side at 1 month postoperatively were significantly lower than those on the affected side preoperatively(P＜0.05).(3)The peak moment of the flexor and extensor muscles and the maximum one-repetition work of the flexor and extensor muscles on the affected side at 3 months postoperatively had no significant difference from those on the unaffected side.The peak flexor-extensor moment of the flexor-extensor muscles on the affected side was higher than that preoperatively(P＜0.05).(4)It is indicated that after unicompartmental knee arthroplasty,the muscle strength on the affected side is poor after 1 month after standard functional rehabilitation.However,after 3 months,there was no significant difference between the affected side and the unaffected side.

Objective To investigate the impact and predictive value of high mobility group protein B1(HMGB1)and angiopoietin like protein 2(Angptl-2)on coronary artery damage in children with Kawasaki disease.Methods A total of 114 children with Kawasaki disease were selected and divided into the normal coronary artery group(85 cases)and the coronary artery lesion group(29 cases)according to the presence or absence of coronary artery lesions.The general clinical data of the two groups were compared.Enzyme linked immunosorbent assay was applied to detect serum levels of HMGB1 and Angptl-2 in children and compared between the two groups.Logistic regression was applied to analyze the influencing factors of coronary artery damage in children with Kawasaki disease.Receiver operating curve(ROC)was applied to analyze the predictive value of serum levels of HMGB1 and Angptl-2 for coronary artery damage in children with Kawasaki disease.Results The serum levels of HMGB1 and Angptl-2 were significantly higher in acute stage than those in remission stage(P＜0.05).Before treatment,the duration of fever,white blood cell count,erythrocyte sedimentation rate,procalcitonin and serum HMGB1 and Angptl-2 levels were significantly higher in the coronary artery lesion group than those in the normal coronary artery group(P＜0.05).Logistic regression analysis showed that the duration of fever,white blood cell count,erythrocyte sedimentation rate,procalcitonin,HMGB1 and Angptl-2 levels before treatment were all independent risk factors for coronary artery damage in children with Kawasaki disease(P＜0.05).The area under the curve(AUC)of serum HMGB1 and Angptl-2 for diagnosing coronary artery damage in children with Kawasaki disease was 0.907(95%CI:0.838-0.953)and 0.857(95%CI:0.780-0.916),respectively,with cut-off value of 59.62 μg/L and 10.35 μg/L,respectively.The AUC,sensitivity and specificity of the combined diagnosis of coronary artery damage in children with Kawasaki disease were 0.958(95%CI:0.903-0.987),93.10%and 90.59%,respectively,which were better than their individual diagnose.Conclusion The serum levels of HMGB1 and Angptl-2 are obviously elevated in children with Kawasaki disease complicated by coronary artery damage,and the combined detection of HMGB1 and Angptl-2 levels in serum can provide a good prediction for coronary artery damage in children with Kawasaki disease.

With the rapid development of generative artificial intelligence(GAI)technologies,their widespread application in academic research and writing is continuously expanding the boundaries of sci-entific inquiry.However,this trend has also raised a series of ethical and regulatory challenges,inclu-ding issues related to authorship,content authenticity,citation accuracy,and accountability.In light of the growing involvement of AI in generating academic content,establishing an open,controllable,and trustworthy ethical governance framework has become a key task for safeguarding research integrity and maintaining trust within the academic community.This expert consensus outlines ethical requirements across key stages of AI-assisted academic writing-including topic selection,data management,citation practices,and authorship attribution.It aims to clarify the boundaries and ethical obligations surrounding AI use in academic writing,ensuring that technological tools enhance efficiency without compromising in-tegrity.The goal is to provide guidance and institutional support for building a responsible and sustainable research ecosystem.

Objective To investigate the impact and predictive value of high mobility group protein B1(HMGB1)and angiopoietin like protein 2(Angptl-2)on coronary artery damage in children with Kawasaki disease.Methods A total of 114 children with Kawasaki disease were selected and divided into the normal coronary artery group(85 cases)and the coronary artery lesion group(29 cases)according to the presence or absence of coronary artery lesions.The general clinical data of the two groups were compared.Enzyme linked immunosorbent assay was applied to detect serum levels of HMGB1 and Angptl-2 in children and compared between the two groups.Logistic regression was applied to analyze the influencing factors of coronary artery damage in children with Kawasaki disease.Receiver operating curve(ROC)was applied to analyze the predictive value of serum levels of HMGB1 and Angptl-2 for coronary artery damage in children with Kawasaki disease.Results The serum levels of HMGB1 and Angptl-2 were significantly higher in acute stage than those in remission stage(P＜0.05).Before treatment,the duration of fever,white blood cell count,erythrocyte sedimentation rate,procalcitonin and serum HMGB1 and Angptl-2 levels were significantly higher in the coronary artery lesion group than those in the normal coronary artery group(P＜0.05).Logistic regression analysis showed that the duration of fever,white blood cell count,erythrocyte sedimentation rate,procalcitonin,HMGB1 and Angptl-2 levels before treatment were all independent risk factors for coronary artery damage in children with Kawasaki disease(P＜0.05).The area under the curve(AUC)of serum HMGB1 and Angptl-2 for diagnosing coronary artery damage in children with Kawasaki disease was 0.907(95%CI:0.838-0.953)and 0.857(95%CI:0.780-0.916),respectively,with cut-off value of 59.62 μg/L and 10.35 μg/L,respectively.The AUC,sensitivity and specificity of the combined diagnosis of coronary artery damage in children with Kawasaki disease were 0.958(95%CI:0.903-0.987),93.10%and 90.59%,respectively,which were better than their individual diagnose.Conclusion The serum levels of HMGB1 and Angptl-2 are obviously elevated in children with Kawasaki disease complicated by coronary artery damage,and the combined detection of HMGB1 and Angptl-2 levels in serum can provide a good prediction for coronary artery damage in children with Kawasaki disease.

Objective To explore the feasibility and safety of non-anticoagulation veno-venous extracorporeal membrane oxygenation(VV-ECMO)in patients with severe chest trauma.Methods A retrospective cohort study method was used.A total of 19 patients with severe chest trauma who received VV-ECMO with a delayed anticoagulation strategy at Zhengzhou Central Hospital Affiliated to Zhengzhou University from January 2018 to October 2021 were included in the delayed anticoagulation group,and 20 patients with severe chest trauma who received VV-ECMO with a non-anticoagulation strategy from November 2021 to October 2024 were included in the non-anticoagulation group.The overall clinical characteristics of the patients were statistically analyzed,including gender,age,injury severity score(ISS),acute physiology and chronic health evaluationⅡ(APACHEⅡ),reason for VV-ECMO,use of vasoactive drugs,oxygenation index(PaO2/FiO2),and interval from injury to VV-ECMO.The primary outcomes were hemorrhagic and thrombotic complications.The secondary outcomes were blood transfusion during VV-ECMO,VV-ECMO time,mechanical ventilation time,intensive care unit(ICU)length of stay,and 28-day mortality.Results There was no significant difference in gender,age,ISS score,APACHEⅡscore,reason for VV-ECMO,use of vasoactive drugs,PaO2/FiO2,and interval from injury to VV-ECMO between the non-anticoagulation group and the delayed anticoagulation group.There was no significant difference in overall incidence of hemorrhagic and thrombotic between the two groups[incidence of hemorrhagic complications:15.0%(3/20)vs.31.6%(6/19),incidence of thrombotic:15.0%(3/20)vs.5.3%(1/19),both P>0.05].The infusion rate of 4 or more paked red blood cell(PRBC)within 24 hours during VV-ECMO in the non-anticoagulation group was significantly lower than that in the delayed anticoagulation group[5.0%(1/20)vs.31.6%(6/19),P<0.05].The amount of PRBC and platelet transfusion and the time on VV-ECMO in the non-anticoagulation group during VV-ECMO were significantly lower than those in the delayed anticoagulation group[PRBC(U):5.8±3.8 vs.8.1±3.1,platelets(U):1(0,1)vs.2(1,3),time on VV-ECMO(hours):71.55±24.37 vs.114.21±34.08,all P<0.05].There were no statistically significant differences in the amount of plasma and cryoprecipitate transfusion during VV-ECMO,mechanical ventilation time,ICU hospitalization time,and 28-day mortality between the two groups.Conclusion For patients with severe chest trauma receiving VV-ECMO withholding routine systemic anticoagulation did not result in thrombotic complications or higher mortality and required less PRBC and platelet transfusions.Non-anticoagulant VV-ECMO is safe and feasible for patients with severe chest trauma with high risk of bleeding.

AIM To investigate the effects of Qinghua Zhixie Formula on improving symptoms of a rat model of diarrhea-predominant irritable bowel syndrome(IBS-D)and its impact on the polarization of M1/M2 macrophages.METHODS The SD rats were randomly divided into the blank group,the model group,the pirenzepine group(20 mg/kg),and the low-dose and high-dose Qinghua Zhixie Formula groups(13.64,27.29 g/kg),with 10 rats in each group.An IBS-D rat model was established using chronic restraint stress combined with gavage of senna leaf decoction.Each group underwent corresponding medication for 14 days.The rats had their changes of food intakes,stool characteristics,and body weight observed;their abdominal withdrawal reflex(AWR)measured to assess the intestinal pain sensitivity;their histopathological changes in the colon mucosa observed using HE staining;their colon mucosa apoptosis assessed using TUNEL staining;their colon expressions of ZO-1,Occludin and Nrf2 proteins detected using immunofluorescence staining;their ratio of M1/M2 macrophages in mesenteric lymph nodes measured by Flow cytometry;and their colon protein expressions of Nrf2,HO-1,p-IκBα and p-P65 analyzed by Western blot.RESULTS Compared with the model group,the groups intervened with Qinghua Zhixie Formula showed increased body weight and AWR pain threshold(P＜0.05,P＜0.01);generally intact colon tissue structure with orderly arrangement of epithelial cells and no significant cell degeneration or shedding;enhanced fluorescence intensity of colon ZO-1 and Occludin proteins(P＜0.05,P＜0.01);reduced apoptosis rate of colon tissue cells(P＜0.05,P＜0.01);more polarized M2 phenotype of macrophages in mesenteric lymph nodes(P＜0.05,P＜0.01);increased protein expressions of colon Nrf2 and HO-1(P＜0.05,P＜0.01);and decreased expressions of p-IKBα/IKBα and p-P65/P65(P＜0.05,P＜0.01).CONCLUSION Qinghua Zhixie Formula can improve the function of the intestinal epithelial mucosal barrier in the IBS-D rat models,and this may be related to the modulation of macrophage polarization toward the M2 phenotype via activation of the Nrf2/HO-1 signaling pathway and inhibited activation of NF-κB signaling pathway.

Objective:To preliminarily explore whether sirtuin1 (SIRT1) activation can inhibit neuronal ferroptosis in rats after traumatic brain injury (TBI) by regulating hypoxia-inducible factor-1α (HIF-1α)-mediated glycolysis.Methods:(1) Six SD rats were randomly divided into sham-operated group and TBI group, with 3 rats in each group; TBI model in the TBI group was established by hydraulic impact method, and rats in the sham-operated group underwent same surgery without impact. Cortical tissues of the two groups were sent for tandem mass tag (TMT) labeled quantitative proteomics detection to analyze the differential expression proteome; Kyoto encyclopedia of genes and genomes (KEGG) and gene set enrichment analysis (GSEA) were used to detect pathway enrichment of the screened differential proteins. (2) Twelve SD rats were randomly divided into sham-operated group and 1-day, 3-day and 7-day post-TBI groups, with 3 rats in each group. Treatment methods were the same as above; Western blotting was used to detect SIRT1 protein expression. (3) Forty-eight rats were randomly divided into sham-operated group, TBI group, TBI+vehicle group and TBI+SIRT1 agonist group, with 12 rats in each group; rats in the sham-operated group and TBI group accepted treatment as above; rats in the TBI+SIRT1 agonist group were intraperitoneally injected with SRT1720 (dissolved in ≤ 5% dimethyl sulfoxide, at a dose of 20 mg/kg) within 30 minutes after modeling, twice a day (with an interval of 12 hours); and rats in the TBI+vehicle group were injected with same dose of dimethyl sulfoxide at the same time. One d after modeling, neurological deficit was assessed using modified Neurological severity score (mNSS), brain water content was measured by dry-wet weight method, histopathological changes in the cortical lesions were observed by HE staining, mitochondrial ultrastructure was examined by transmission electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the brain tissues were detected by colorimetry, and protein expressions of SIRT1, HIF-1α (key protein in the glycolytic pathway), glutathione peroxidase 4 (GPX4, key protein in the ferroptosis pathway), and acyl-CoA synthetase long-chain family member 4 (ACSL4, key protein in the ferroptosis pathway) were evaluated by Western blotting.Results:(1) KEGG analysis revealed that the glycolysis pathway and HIF-1 signaling pathway were obviously enriched in the cortical tissues of rats in the TBI group compared with the sham-operated group; GSEA showed that the HIF-1 signaling pathway (mmu04066) and ferroptosis pathway (mmu04216) gene sets in the cortical tissues of rats in the TBI group exhibited enrichment trends compared with those in the sham-operated group. (2) Compared with the sham-operated group, the 1-day, 3-day, and 7-day post-TBI groups had significantly decreased SIRT1 protein expression ( P<0.05), with the most prominent decline in 1-day post-TBI group. (3) Compared with the TBI+vehicle group, rats in the TBI+SIRT1 agonist group showed significantly reduced mNSS score and brain tissue water content (9.83±1.17 vs. 7.66±1.21; [83.62±0.91]% vs. [80.09±0.68]%, P<0.05). HE staining indicated clearer structure of the cortical area at the injury sites, and improved neuron morphology in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group; and transmission electron microscopy showed reduced mitochondrial shrinkage and partial restoration of cristae structures in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group. Compared with the TBI+vehicle group, the TBI+SIRT1 agonist group exhibited significantly decreased MDA content ([62.72±9.20] nmol/g vs. [39.34±3.48] nmol/g), increased SOD activity ([1.95±0.23] U/mg vs. [2.48±0.14] U/mg), elevated GPX4 protein expression (0.37±0.04 vs. 0.46±0.03), and decreased HIF-1α and ACSL4 protein expressions (1.16±0.15 vs. 0.81±0.12; 1.14±0.06 vs. 1.29±0.04), with significant differences ( P<0.05). Conclusion:SIRT1 activation can exert neuroprotective effect by inhibiting HIF-1α-mediated glycolysis and reducing neuronal ferroptosis after TBI.

Objective:To investigate the effects and underlying mechanism of ionizing radiation on the adipogenic of mesenchymal stem cells(MSCs).Methods:Mouse MSCs were cultured in vitro and treated with 2 Gy and 6 Gy radiation with 60Co,and the radiation dose rate was 0.98 Gy/min.Bulk RNA-seq was performed on control and irradiated MSCs.The changes of adipogenic differentiation and oxidative stress pathways of MSC were revealed by bioinformatics analysis.Oil Red O staining was used to detect the adipogenic differentiation ability of MSCs in vitro,and real-time fluorescence quantitative PCR(qPCR)was used to detect the expression differences of key regulatory factors Cebpa,Lpl and Pparg after radiation treatment.At the same time,qPCR and Western blot were used to detect the effect of inhibition of Nrf2,a key factor of antioxidant stress pathway,on the expression of key regulatory factors of adipogenesis.Moreover,the species conservation of the irradiation response of human bone marrow MSCs and mouse MSC was determined by qPCR.Results:Bulk RNA-seq suggested that ionizing radiation promotes adipogenic differentiation of MSCs and up-regulation of oxidative stress-related genes and pathways.The results of Oil Red O staining and qPCR showed that ionizing radiation promoted the adipogenesis of MSCs,with high expression of Cebpa,Lpl and Pparg,as well as oxidative stress-related gene Nrf2.Nrf2 pathway inhibitors could further enhance the adipogenesis of MSCs in bone marrow after radiation.Notably,the similar regulation of oxidative pathways and enhanced adipogenesis post irradiation were observed in human bone marrow MSCs.In addition,irradiation exposure led to up-regulated mRNA expression of interleukin-6 and down-regulated mRNA expression of colony stimulating factor 2 in human bone marrow MSCs.Conclusion:Ionizing radiation promotes adipogenesis of MSCs in mice,and oxidative stress pathway participates in this effect,blocking Nrf2 further promotes the adipogenesis of MSCs.Additionally,irradiation activates oxidative pathways and promotes adipogenic differentiation of human bone marrow MSCs.

Objective:To establish an in vitro cell model simulating acute graft-versus-host disease(aGVHD)bone marrow microenvironment injury with the advantage of mouse serum of aGVHD model and explore the effect of serum of aGVHD mouse on the adipogenic differentiation ability of mesenchymal stem cells(MSCs).Methods:The 6-8-week-old C57BL/6N female mice and BALB/c female mice were used as the donor and recipient mice of the aGVHD model,respectively.Bone marrow transplantation(BMT)mouse model(n=20)was established by being injected with bone marrow cells(1 × 10'per mouse)from donor mice within 4-6 hours after receiving a lethal dose(8.0 Gy,72.76 cGy/min)of y ray general irradiation.A mouse model of aGVHD(n=20)was established by infusing a total of 0.4 ml of a mixture of donor mouse-derived bone marrow cells(1 × 107 per mouse)and spleen lymphocytes(2 × 106 per mouse).The blood was removed from the eyeballs and the mouse serum was aspirated on the 7th day after modeling.Bone marrow-derived MSCs were isolated from 1-week-old C57BL/6N male mice and incubated with 2％,5％and 10％BMT mouse serum and aGVHD mouse serum in the medium,respectively.The effect of serum in the two groups on the in vitro adipogenic differentiation ability of mouse MSCs was detected by Oil Red O staining.The expression levels of related proteins PPARy and CEBPα were detected by Western blot.The expression differences of key adipogenic transcription factors including PPARy,CEBPα,FABP4 and LPL were determined by real-time quantitative PCR(RT-qPCR).Results:An in vitro cell model simulating the damage of bone marrow microenvironment in mice with aGVHD was successfully established.Oil Red O staining showed that the number of orange-red fatty droplets was significantly reduced and the adipogenic differentiation ability of MSC was impaired at aGVHD serum concentration of 10％compared with BMT serum.Western blot experiments showed that adipogenesis-related proteins PPARy and CEBPα expressed in MSCs were down-regulated.Further RT-qPCR assay showed that the production of PPARy,CEBPα,FABP4 and LPL,the key transcription factors for adipogenic differentiation of MSC,were significantly reduced.Conclusion:The adipogenic differentiation capacity of MSCs is inhibited by aGVHD mouse serum.