1.Effect evaluation of pharmaceutical service based on root cause analysis combined with KAP theory on decreasing the protocol violations of investigational medicinal products rate in pediatric clinical trials
Chunyan GUO ; Yi ZHANG ; Yuxin YANG ; Yuguang LIANG ; Qian DING ; Qian WANG ; Chengyue ZHANG ; Yu SUN ; Peng GUO
China Pharmacy 2026;37(9):1206-1210
OBJECTIVE To investigate the effect of pharmaceutical services guided by root cause analysis (RCA) in a problem-oriented manner combined with knowledge-attitude-practice (KAP) theory on reducing the incidence of protocol violations of investigational medicinal products in pediatric clinical trials. METHODS A total of 617 participants from 69 drug clinical trial projects conducted in our hospital from January 2016 to December 2020 were selected as the control group, and 868 participants from 72 drug clinical trial projects from January 2022 to December 2025 as the observation group. RCA was performed on the protocol violations of investigational medicinal product in the control group to identify the types and underlying causes. The control group received routine pharmaceutical services for drug clinical trials, while the observation group was provided with precision pharmaceutical services from the three dimensions of knowledge, attitude and practice on the basis of routine pharmaceutical services, according to the root causes identified by RCA. The occurrence of investigational medicinal products protocol violations was compared between the two groups. RESULTS The total incidence of protocol violations of investigational medicinal products, as well as the incidences of minor and major protocol violations, were all significantly lower in the observation group than in the control group ( P <0.001). The main types of protocol violations in both groups included missed/under-/over-dosing of medications, non-adherence to administration time, failure to adjust dosage as required, and combined medication/vaccination in violation of the protocol. Regarding the responsible subjects of protocol violations, the incidences of protocol violations attributed to participants and their guardians as well as investigators and accidental factors were significantly lower in the observation group than in the control group ( P <0.001, P <0.001, P =0.025). However, there were no statistically significant differences in the incidences of protocol violations caused by sponsor-related reasons between the two groups ( P >0.05). CONCLUSIONS Pharmaceutical services led by pharmacists, based on problem-oriented RCA and combined with KAP theory, can effectively reduce the protocol violations of investigational medicinal products rate in pediatric clinical trials, thereby safeguarding the safety and rights of study participants.
2.Study of adsorption of coated aldehyde oxy-starch on the indexes of renal failure
Qian WU ; Cai-fen WANG ; Ning-ning PENG ; Qin NIE ; Tian-fu LI ; Jian-yu LIU ; Xiang-yi SONG ; Jian LIU ; Su-ping WU ; Ji-wen ZHANG ; Li-xin SUN
Acta Pharmaceutica Sinica 2025;60(2):498-505
The accumulation of uremic toxins such as urea nitrogen, blood creatinine, and uric acid of patients with renal failure
3.Screening key genes of PANoptosis in hepatic ischemia-reperfusion injury based on bioinformatics
Lirong ZHU ; Qian GUO ; Jie YANG ; Qiuwen ZHANG ; Guining HE ; Yanqing YU ; Ning WEN ; Jianhui DONG ; Haibin LI ; Xuyong SUN
Organ Transplantation 2025;16(1):106-113
Objective To explore the relationship between PANoptosis and hepatic ischemia-reperfusion injury (HIRI), and to screen the key genes of PANoptosis in HIRI. Methods PANoptosis-related differentially expressed genes (PDG) were obtained through the Gene Expression Omnibus database and GeneCards database. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA) were used to explore the biological pathways related to PDG. A protein-protein interaction network was constructed. Key genes were selected, and their diagnostic value was assessed and validated in the HIRI mice. Immune cell infiltration analysis was performed based on the cell-type identification by estimating relative subsets of RNA transcripts. Results A total of 16 PDG were identified. GO analysis showed that PDG were closely related to cellular metabolism. KEGG analysis indicated that PDG were mainly enriched in cellular death pathways such as apoptosis and immune-related signaling pathways such as the tumor necrosis factor signaling pathway. GSEA results showed that key genes were mainly enriched in immune-related signaling pathways such as the mitogen-activated protein kinase (MAPK) signaling pathway. Two key genes, DFFB and TNFSF10, were identified with high accuracy in diagnosing HIRI, with areas under the curve of 0.964 and 1.000, respectively. Immune infiltration analysis showed that the control group had more infiltration of resting natural killer cells, M2 macrophages, etc., while the HIRI group had more infiltration of M0 macrophages, neutrophils, and naive B cells. Real-time quantitative polymerase chain reaction results showed that compared with the Sham group, the relative expression of DFFB messenger RNA in liver tissue of HIRI group mice increased, and the relative expression of TNFSF10 messenger RNA decreased. Cibersort analysis showed that the infiltration abundance of naive B cells was positively correlated with DFFB expression (r=0.70, P=0.035), and the infiltration abundance of M2 macrophages was positively correlated with TNFSF10 expression (r=0.68, P=0.045). Conclusions PANoptosis-related genes DFFB and TNFSF10 may be potential biomarkers and therapeutic targets for HIRI.
4.Icariin pretreatment enhances effect of human periodontal stem cells on M1-type macrophages
Ting YU ; Dongmei LYU ; Hao DENG ; Tao SUN ; Qian CHENG
Chinese Journal of Tissue Engineering Research 2025;29(7):1328-1335
BACKGROUND:Human periodontal stem cells have a certain inhibitory effect on the pro-inflammatory function of M1-type macrophages,and it is not clear whether icariin,which has anti-inflammatory and other pharmacological activities,can enhance the inhibitory effect of human periodontal stem cells on M1-type macrophages. OBJECTIVE:To investigate the effect of icariin on M1 macrophages after pretreatment of human periodontal stem cells. METHODS:Primary human periodontal stem cells were isolated,cultured and characterized.THP-1 was induced and M1-type macrophages were identified by immunofluorescence staining and PCR.Human periodontal stem cells were cultured with α-MEM complete medium containing concentrations of 10-7,10-6,10-5,and 10-4 mol/L icariin,and the cytotoxicity of Icariin on human periodontal stem cells was detected by the CCK-8 assay at 1,3,5,and 7 days,respectively.α-MEM complete medium,untreated α-MEM conditioned medium for human periodontal stem cells and α-MEM conditioned medium for human periodontal stem cells pretreated with icariin for 24 hours were conditioned with RPMI-1640 complete medium in a 1:1 ratio for M1-type macrophages in the control,untreated,and pretreated groups,and 24 hours later,the mRNA expression of inflammatory factors in M1 macrophages was detected by RT-PCR.The protein expression of inflammatory factors in M1 macrophages was detected by ELISA.The expression of surface markers and nuclear factor-κB pathway-related proteins in M1/M2 macrophages was detected by western blot assay. RESULTS AND CONCLUSION:(1)CCK-8 assay results showed that 10-7,10-6,10-5,10-4 mol/L icariin was not cytotoxic to the human periodontal stem cells,and from day 5 onwards,all the concentrations increased the cell viability,and promoted the cell proliferation.10-4 mol/L icariin was selected for follow-up experiment.(2)RT-PCR and ELISA results showed that compared with the control group,the untreated group and the pretreated group both decreased the expression and secretion of interleukin-1β,interleukin-6,and tumor necrosis factor-α of M1-type macrophages(P<0.05),and the pretreated group was lower than the untreated group(P<0.05).(3)Western blot assay results showed that compared with the untreated group,the expression of CD86 was significantly lower in the pretreated group(P<0.05);compared with the control group,the expression of CD206,a surface marker of M2-type macrophages,was elevated in both the untreated and pretreated groups(P<0.01),and it was significantly higher in the pretreated group than in the untreated group(P<0.01).In M1-type macrophages after 24 hours of conditioned culture,compared with the control group,the expression of nuclear factor-κB/P65 was decreased in the untreated group and the pretreated group(P<0.01),and the expression of p-IκBα was decreased only in the pretreated group(P<0.01);the expression of both nuclear factor-κB/P65 and p-IκBα was significantly reduced in the pretreated group compared with the untreated group(P<0.05),while the difference of IκBα in the three groups was not statistically significant.(4)These results indicated that icariin enhanced the inhibitory effect of human periodontal stem cells on M1-type macrophages,and this effect may be related to the inhibition of the nuclear factor-κB signaling pathway of macrophages.
5.Efficacy Mechanism of Xianlian Jiedu Prescription Against Colorectal Cancer Recurrence vias Regulating Angiogenesis
Yanru XU ; Lihuiping TAO ; Jingyang QIAN ; Weixing SHEN ; Jiani TAN ; Chengtao YU ; Minmin FAN ; Changliang XU ; Yueyang LAI ; Liu LI ; Dongdong SUN ; Haibo CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):79-87
ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer (CRC) and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit, followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group, low-dose Xianlian Jiedu prescription (XLJDP-L) group (6.45 g·kg-1·d-1), medium-dose Xianlian Jiedu prescription (XLJDP-M) group (12.9 g·kg-1·d-1), high-dose Xianlian Jiedu prescription (XLJDP-H) group (25.8 g·kg-1·d-1), and 5-fluorouracil (5-FU) group (1×10-3 g·kg-1·d-1). The mice were euthanized after 14 days of continuous intervention, and recurrent tumor tissue was harvested. Hematoxylin and eosin (HE) staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry (IHC) was employed to assess the expression of proliferating cell nuclear antigen (Ki67), vascular endothelial growth factor (VEGF), and platelet-endothelial cell adhesion molecule (CD31) in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 (ANG-2), VEGF, phosphorylated-protein kinase B (p-Akt), protein kinase B (Akt), phosphorylated-phosphatidylinositol 3-kinase (p-PI3K), and phosphatidylinositol 3-kinase (PI3K) in recurrent tumor tissue. ResultsBefore treatment, there were no statistical differences in tumor volume, tumor weight, and body mass among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group, indicating model stability. After treatment, compared with those in the model group, the tumor volume and tumor weight in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01), showing dose dependency. Meanwhile, there were no significant differences in body weight among the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group, tumor tissue in the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group had loosely arranged cells, increased intercellular spaces, small and shriveled nuclei, light staining, fewer mitotic figures and atypical nuclei, and increased necrotic areas. IHC showed that compared with those of the model group, the positive rates of Ki67, VEGF, and CD31 in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly reduced (P<0.01) in a dose-dependent manner. Western blot results showed that compared with those of the model group, the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L, XLJDP-M, and XLJDP-H groups and the 5-FU group were significantly downregulated (P<0.05, P<0.01), and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner (P<0.05, P<0.01). ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2, VEGF, and CD31.
6.Efficacy of remimazolam-based anesthesia in daytime laparoscopic cholecystectomy: a multicenter, non-inferiority, randomized controlled trial
Qian HU ; Jian GUO ; Haijun YUAN ; Weiping LEI ; Haipeng LIU ; Chen YONG ; Yanhui HU ; Junping CHEN ; Jianliang SUN ; Zhijian LAN ; Jianhong XU ; Xin YU ; Gang CHEN
Chinese Journal of Anesthesiology 2025;45(6):720-725
Objective:To evaluate the efficacy of remimazolam-based anesthesia in daytime laparoscopic cholecystectomy.Methods:In this multicenter, non-inferiority, randomized controlled trial, 300 American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ patients of either sex, aged 18-60 yr, with body mass index of 18-28 kg/m 2, who underwent daytime laparoscopic cholecystectomy under general anesthesia with tracheal intubation at Sir Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine, the Fourth Affiliated Hospital of Zhejiang University School of Medicine, Jinhua Hospital Affiliated to Zhejiang University School of Medicine, Hangzhou First People′s Hospital Affiliated to Westlake University School of Medicine, Ningbo No. 2 Hospital, and the Second Affiliated Hospital of Nanchang University from August 2021 to August 2023, were selected and divided into 2 groups ( n=150 each) using a random number table method: remimazolam group (R group) and propofol group (P group). Anesthesia was induced as follows: Sufentanil was intravenously injected at a rate of 0.5 μg/kg, remimazolam was intravenously injected at a rate of 0.3 mg/kg in group R, propofol was intravenously injected at a rate of 2.0-2.5 mg/kg in group P, and cisatracurium besilate was intravenously injected at a rate of 0.2 mg/kg after loss of consciousness in two groups. The patients were mechanically ventilated after tracheal intubation. Anesthesia was maintained as follows: Remimazolam was intravenously injected at a rate of 0.5-1.0 mg·kg -1·h -1 in group R, propofol was intravenously injected at a rate of 4-10 mg·kg -1·h -1 in group P, and remifentanil was intravenously infused at a rate of 0.25-2.00 μg·kg -1·min -1, maintaining intraoperative bispectral index value of 40-60. The success rate of sedation was recorded, and non-inferiority tests were conducted. The time to loss of consciousness, emergence time, extubation time, recovery time of orientation, time of stay in post-anesthesia care unit and occurrence of delayed emergence were recorded. Liver function and renal function were measured before operation and within 24 h after operation. The occurrence of abnormal alanine transaminase, abnormal aspartate transaminase, abnormal creatinine and abnormal urea was recorded. The occurrence of adverse reactions during and after operation was recorded. Results:The success rates of sedation were 98.6% and 99.3% in group R and group P, respectively, there was no statistically significant difference in the success rate of sedation between the two groups ( P>0.05), and the difference in the success rates of sedation between the two groups was -0.007 (95% confidence interval-0.0301-0.0161), which met the pre-set non-inferiority criteria(95% confidence interval >-0.055). Compared with group P, the time to loss of consciousness and recovery time of orientation were significantly prolonged, and the incidence of delayed emergence was increased ( P<0.05), and no statistically significant changes were found in the emergence time, extubation time, time of stay in post-anesthesia care unit and severity of postoperative nausea and vomiting in group R ( P>0.05). There was no statistically significant difference in the abnormal rates of alanine transaminase, aspartate transaminase, creatinine and urea before and after operation between the two groups ( P>0.05). Conclusions:The efficacy of remimazolam-based anesthesia in daytime laparoscopic cholecystectomy is not inferior to that of propofol-based anesthesia.
7.Proficiency testing for 11 clinical biobanks in Beijing City: simulation study and result analysis
Qian ZHANG ; Yun ZHANG ; Lu HAN ; Min LIU ; Yongbo YU ; Yan WANG ; Ying HU ; Hui ZHONG ; Dan GUO ; Shipeng SUN ; Jinxi LIN ; Siyuan XU ; Xiaokun TANG ; Gaoyuan SUN ; Chuanbao ZHANG ; Hexin LI
Chinese Journal of Preventive Medicine 2025;59(9):1590-1596
Objective:To evaluate the sample preparation proficiency and storage proficiency of 11 clinical biobanks in Beijing through simulated experiments, and to establish an assessment method for the quality comparability of biological samples.Methods:An exploratory research design was adopted. In November 2023, artificial composite serum quality control materials containing six recombinant human protein markers—recombinant human alanine aminotransferase (rhALT), recombinant human aspartate aminotransferase (rhAST), recombinant human creatine kinase (rhCK), recombinant human creatine kinase-MB (rhCK-MB), recombinant human B-type natriuretic peptide (rhBNP), and recombinant human troponin I (rhTNI)—were distributed to 11 clinical biobanks in Beijing City. Sample preparation and storage followed the standardized operating procedures. Proficiency differences were assessed through statistical analysis.Results:Three-way repeated measures ANOVA revealed all six protein markers showed a declining trend over storage time in ultra-low-temperature environments ( F values 11.68-4 179.66, all P<0.01). However, neither long-term/temporary refrigerator types ( F values 0.01-1.23, all P>0.05)nor placement locations within refrigerators significantly affected the stability of these six proteins ( F valus 0.03-1.47, all P>0.05). The biases in detection results for rhALT, rhAST, rhTNI, and rhBNP at different storage time points were within the allowable bias limits for each item, supporting their use as markers for protein stability in biobank samples. All 11 institutions passed the storage proficiency assessment. In the preparation proficiency assessment, deviations were observed in post-preparation sample results, with a notably high out-of-control rate for rhCK (36.36%). Conclusion:Sample preparation proficiency can serve as a quality control metric for clinical biobanks. Future external quality assessment systems for biobanks should focus on sample preparation rather than storage processes.
8.A single Kocher-Langenbeck approach to improve anterior column screw fixation for transverse + posterior wall acetabular fractures
Hongtao LI ; Longxin AN ; Qian WU ; Jiazhi GAO ; Bin YU ; Jun LIU ; Xuecheng SUN
Chinese Journal of Orthopaedic Trauma 2025;27(6):536-540
Objective:To investigate the efficacy of using a single Kocher-Langenbeck approach to improve anterior column screw fixation for the treatment of transverse + posterior wall acetabular fractures.Method:A retrospective analysis was conducted of the data of 21 patients who had been treated with a single Kocher-Langenbeck approach to improve anterior column screw fixation for transverse + posterior wall acetabular fractures at Department of Orthopaedics, Weifang People's Hospital from May 2021 to May 2023. There were 11 males and 10 females with an age of (51.3±15.5) years. The time from injury to surgery was (10.4±3.6) days. The anterior column screw fixation was improved as follows: screws were implanted 1 to 2 cm in front of the apex of the great notch of the external iliac plate of the pelvis to penetrate out at the upper branch root of the pubic ramus, with the whole screw fully located within the bony channel. Such indexes were recorded as surgical time, intraoperative blood loss, intraoperative fluoroscopy frequency for improved placement of anterior column screws, fracture healing time, incidence of complications, and fracture reduction and hip joint function at the last follow-up.Results:For this cohort, surgical time was (120.9±40.7) minutes, intraoperative blood loss (330.2±65.9) mL, and intraoperative fluoroscopy frequency for improved placement of anterior column screws (6.2±2.3) times. The 21 patients were followed up for (21.5±5.9) months. All fractures achieved bony union after (3.2±1.1) months. Incision fat liquefaction occurred in 1 patient after surgery and symptoms of sciatic nerve injury were observed in 2 patients. Follow-ups observed no such complications as loosening or breakage of internal fixators. According to the Matta scoring criteria, the quality of fracture reduction at the last follow-up was evaluated as anatomical reduction in 15 cases, as satisfactory reduction in 4 cases, as acceptable reduction in 2 cases, giving a satisfaction rate of 90.5% (19/21). According to the improved Merle d'Aubigné & Poster scoring criteria, hip function was evaluated as excellent in 16 cases, as good in 2 cases, and as fair in 3 cases, giving an excellent and good rate of 85.7% (18/21).Conclusion:It is effective to use a single Kocher-Langenbeck approach to improve anterior column screw fixation for the treatment of transverse + posterior wall acetabular fractures, showing advantages of reduced surgical time and intraoperative bleeding.
9.Clinicopathological analysis of 495 cases of gastrointestinal mesenchymal tumors resected via endoscopy
Shuaixia YU ; Yao HUANG ; Xiao HU ; Jing FU ; Huajun SUN ; Baijie TANG ; Qian TANG ; Ying XU ; Xudan YANG
Chinese Journal of Clinical and Experimental Pathology 2025;41(7):897-903
Purpose To characterize the clinicopathological features of gastrointestinal mesenchymal tumors(GMTs)resected via endoscopic techniques.Methods A retrospective analysis was conducted on 495 cases of endo-scopically resected GMTs.Clinical information,histomorphological findings,immunohistochemical profiles,and mo-lecular characteristics were reviewed.Results The cohort included 495 patients aged 20-78 years(median:53 years).The majority of tumors were located in the stomach(58.8%)and esophagus(36.8%).Histologically,most tumors consisted of spindle cells,with a minority composed of epithelioid cells;fibrocollagenous or myxoid stroma was occasionally observed.Immunohistochemically,leiomyomas showed diffuse positivity for α-SMA(98.8%)and desmin(99.3%),gastrointestinal stromal tumors(GISTs)expressed CD117(99.4%),DOG1(97.6%),and CD34(97.0%),and schwannomas were positive for S-100(93.7%).The predominant tumor types were leiomyomas(54.1%)and GISTs(33.7%),while the remaining 12.2%comprised other rare types.Various endoscopic resection techniques were employed based on the tumors' anatomical depth,including endoscopic submucosal dissection(ESD,40.5%),submucosal tunneling endoscopic resection(STER,17.1%),endoscopic mucosal resection(EMR,16.5%),endoscopic full-thickness resection(EFTR,13.9%),and endoscopic submucosal excavation(ESE,12.0%).EMR and ESD were primarily used for superficial lesions,while deeper tumors with were more often treated with STER,EFTR,and ESE.The rate of negative resection margins was lower in GISTs(72.2%)and other tumors with indistinct margins,compared to leiomyomas(92.6%)and those with well-defined boundaries.Conclusion Leiomyomas and GISTs are the most common gastrointestinal mesenchymal tumors resected via endoscopy.A variety of resection techniques are applicable depending on tumor location and depth.Accurate pathological diagnosis should be based on HE morphology,supplemented by endoscopic findings,margin status,immunohistochemistry,and necessary molecular tests.
10.Effects of aerobic exercise via Gut-Muscle Axis on sarcopenia and inflammation in CT26 tumor mice
Shuling ZHANG ; Junwei WANG ; Yu QIAN ; Shiliang HU ; Junzhi SUN ; Chun WANG
Chinese Journal of Sports Medicine 2025;44(6):466-482
Objective To explore the effect of aerobic exercise on sarcopenia in CT26 tumor mice through the gut microbiota-skeletal muscle axis and its potential mechanism.Methods Forty-eight SPF BALB/c male mice aged 8 weeks were randomly divided into a normal control group(group NC,n=12),a normal control+aerobic exercise group(group NA,n=12),a CT26 tumor-associated sarco-penia group(group TS,n=12)and a CT26 tumor-associated sarcopenia+aerobic exercise group(group TSA,n=12).After a 2-week adaptive exercise period,CT26 cell suspension(0.2 mL,1×107 cells/mL)was subcutaneously injected into the dorsal region of mice in group TS and group TSA,while group NC and group NA were inoculated with 0.2 mL normal saline.One day later,group NA and group TSA underwent a 4-week aerobic treadmill running intervention(14 m/min,60 min/day,6 days/week).If a mouse failed to complete the target intensity on a given day,its running speed was reduced by 20%the following day.Moreover,group NA and group TSA received no exercise interven-tion.General health status of all groups was monitored throughout the study.The forelimb gripping force,rotarod walking time and the cross-sectional area of gastrocnemius muscle fibers were measured.Moreover,the ultrastructural changes of gastrocnemius were observed by transmission electron micro-scope,while the cecum contents were observed by 16S rRNA sequencing.Meanwhile,the pathological morphological changes of small intestine wall were observed using HE,while the contents of tumor ne-crosis factor-α(TNF-α),interleukin-6(IL-6),interleukin-1β(IL-1β)and lipopoly saccharide(LPS)of serum and gastrocnemius muscle were determined by using ELISA.Moreover,small intestine zonula occludens-1(ZO-1),Occludin and muscle ring finger 1(MuRF1),muscle atrophy F-box(MAF-bx),Toll-like receptor 4(TLR4)/myeloid differentiation factor 8(MyD88)/nuclear transcription factor-κB(NF-κB)signal pathway related proteins expression levels of gastrocnemius muscle,as well as muscle ring finger 1(MuRF1)and muscle atrophy F-box(MAF-bx)were detected using Western blot.Results(1)Subcutaneous tumors were prominent in group TS.The forelimb gripforce,wet weight and cross-sectional area of gastrocnemius,and rotarod walking time in group TS at the 5th and 6th weekends were significantly lower than group NC(P<0.01),while the relative expression levels of MuRF1 and MAF-bx proteins in the gastrocnemius muscle in group TS were significantly higher than the latter group(P<0.05,P<0.01).Moreover,transmission electron microscopy revealed sparse and dis-organized muscle fiber arrangement in group TS,with visible Z-lines but indistinct M-lines,as well as shortened,poorly defined,and thin sarcomeres.(2)The group TSA exhibited significantly higher α-diversity indices(Shannon,Chao1,and Observed species)in gut microbiota compared to group TS(P<0.05,P<0.01,P<0.05),while the β-diversity differed to some extent between the two groups.At phylum level,the F/B ratio was lower in group NA than that group NC,with that of group TSA low-er than group TS.At genus level,the relative abundances of Muribaculaceae,Bacteroides,Roseburia,and Alistipes in group NA were higher than group NC,and those of Alloprevotellaand Helicobacter were significantly lower,while those of Muribaculaceae,Lachnospiraceae_NK4A136_group,Roseburia,Bacteroides,Lachnospiraceae_UCG-001 of group TSA were significantly higher than group TS.Nota-bly,Roseburia and Alistipes were the most significantly enriched genera in group TSA and group TS(LDA score>4 for both).(3)The inflammatory cells of the small intestine wall in group TSA signifi-cantly reduced than those in group TS(P<0.05).Moreover,the expression level of ZO-1 proteins in the small intestine wall were significantly higher in group NA than group NC,and that of ZO-1 and Occludin proteins in the small intestine wall was significantly higher in group TSA than those in group TS(P<0.05).(4)The group NA exhibited significantly lower LPS concentrations in the gastrocne-mius muscle compared to group NC(P<0.05).Similarly,the group TSA showed reduced LPS levels in both serum and gastrocnemius compared to group TS(P<0.05).The relative protein expression of p-IκBα and p-NF-κB p65 significantly decreased in group TSA compared to group TS(P<0.05).The group TS displayed elevated concentrations of pro-inflammatory cytokines(TNF-α,IL-1β)in serum and gastrocnemius compared to group NC(P<0.01,P<0.05).The group TSA demonstrated significantly lower levels of TNF-α and IL-6 in these tissues in comparison with group TS(P<0.05,P<0.01).Conclusion Aerobic exercise can improve gut microbiota α diversity,adjusting its composition,improv-ing intestinal mucosal barrier function,reducing the LPS-induced pro-inflammatory response,and delay-ing skeletal muscle atrophy.The underlying mechanism may involve down regulation of TLR4/MyD88/NF-κB signaling in skeletal muscle,which has a positive effect on Gut-Muscle Axis.

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