Animal experimentation constitutes a critical link between basic research and clinical application, making its research quality and translational efficiency paramount. Although considerable progress has been made in standardizing operational procedures and ethical guidelines, the standardization of outcome evaluation systems has significantly lagged, creating a key bottleneck that constrains the quality of biomedical research and evidence synthesis. This deficiency is manifested by pronounced heterogeneity in outcome selection across similar studies, incomplete methodological reporting, and disparate criteria for result interpretation, which severely impairs the comparability of findings and the evidence integration. To cope with this challenge, this paper systematically introduces a mature methodological tool from clinical research–the core outcome set (COS)–and explores its construction strategies and application potential in the field of animal experimentation. Given the extensive diversity of animal experiments, a pragmatic strategy of "focusing on key areas, implementing phased pilots, and promoting gradual expansion" should be adopted. This approach prioritizes the development of domain-specific COS for disease areas characterized by high research volume, urgent translational needs, and well-established animal models. A multi-source integration pathway for COS development is detailed, comprising systematic literature searches, methodological appraisals, and expert consensus, with the feasibility of leveraging artificial intelligence (AI) to enhance efficiency also being examined. The development and promotion of such COS are not intended to restrict scientific exploration; rather, they aim to establish a new, tiered evaluation paradigm consisting of "core outcomes" (mandatory), "recommended outcomes" (encouraged), and "exploratory outcomes" (optional). This framework is expected not only to enhance research quality through standardization and to adhere to the "3R" principles but also to accelerate the accumulation of high-quality evidence. This, in turn, provides a solid foundation for higher-level evidence synthesis, ultimately facilitating the effective translation of basic research findings into clinical practice and providing an essential methodological framework for scientific advancement in relevant disciplines.

ObjectiveBased on 16S rDNA technology and molecular biology methods， the molecular mechanism of Shenling Baizhusan in the treatment of diarrhea-predominant irritable bowel syndrome （IBS-D） was investigated. MethodsThe 42 SD rats with SPF were randomly divided into no control group， SLBZS-H， medium （SLBZS-M）， low （SLBZS-L） dose group， positive control group and model group， with 7 rats in each group. The rat model of IBS-D was prepared by ice-cold senna （0.45 g∙mL^-1） gavage （10 mL∙kg^-1） combined with restraint stress for 14 consecutive days. After successful modeling， the corresponding drugs were given to each group with a gavage volume of 10 mL∙kg^-1： The positive group was administered with 2.36 ， 1.18， 0.59 g∙mL^-1 of Shenling Baizhusan in the Positive group and the Model group with the same volume of normal saline for 14 d. The general condition of the rats： Weight， feces， mental state and death were observed and recorded. The body weight， abdominal wall retraction reflex score （AWR） and loose stool rate of rats in each group were measured before （the first day）， after the model （day 14） and after treatment （day 28）. Hematoxylin-eosin staining was used to observe the morphological characteristics of colon tissues of experimental animals. Enzyme-linked immunosorbent assay was used to quantitatively analyze the concentration of inflammatory mediators in the peripheral blood of experimental animals. Western blotting was used to detect the expression levels of key proteins of Toll-like receptor 4 （TLR4）， Toll-like receptor 2 （TLR2）， myeloid differentiation factor 88 （MyD88） and nuclear factor-κB （NF-κB） signaling pathway in rat colon tissue. 16S rDNA technology was used to detect the structural changes of intestinal microbiota in rats. ResultsCompared with Control， the colon of the Model group showed partial mucosal epithelial shedding and inflammatory cell infiltration. The contents of TNF-α， IL-1β， IL-6 and 5-HT in serum increased （P<0.05）， the protein expressions of TLR2， TLR4， MyD88 and NF-κB in colon tissue increased （P<0.05）， the diversity indices of Richness， Chao1， abundance-based coverage estimator（ACE） and Shannon decreased （P<0.05）， and the phylum Firmicutes， Actinobacteria， The relative richness of Bacteroides-H， Lactobacillus and Ligilactobacillus decreased （P<0.05）， while the relative richness of Bacteroidetes， Proteobacteria and Prevotella increased （P<0.05）. Compared with the model group， the colonic structure and organization of the SLBZS-H group， SLBZS-M group， SLBZS-L group and Positive group were clearer， and only a small number of inflammatory cells were present in some areas， and the serum contents of TNF-α， IL-1β， IL-6 and 5-HT were decreased （P<0.05）， TLR2， TLR4， The protein expressions of MyD88 and NF-κB decreased （P<0.05）， and compared with the model group， the diversity indices of Richness， Chao1， ACE and Shannon in the SLBZS-H， SLBZS-M and SLBZS-L groups increased （P<0.05）， and the richness of Firmicutes and Actinobacteria increased （P<0.05）. The richness of Proteobacteria and Prevotella decreased （P<0.05）， and the abundance of Prevotella decreased （P<0.05）， Bacteroides-H， Muribaculum， Lactobacillus and salivarius in the Positive group salivarius （P<0.05）. ConclusionShenling Baizhusan can effectively treat IBS-D， and its molecular mechanism may be to play a therapeutic role by improving intestinal flora and inhibiting the TLRS/MyD88/NF-κB signaling pathway to reduce inflammatory response.

Animal experiments are essential tools in biomedical research, serving as a bridge between basic research and clinical trials. Systematic reviews and meta-analyses (SRs/MAs) of animal experiments are crucial methods for integrating evidence from animal experiment, which can facilitate the translation of findings into clinical research, reduce translational risks, and promote resource integration in basic research. With the continuous development of the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) methodology, its application in SRs/MAs of animal experiments has gained increasing attention. This article first outlines the principles and specific applications of the GRADE methodology in SRs/MAs of animal experiments, including qualitative descriptive systematic reviews, meta-analyses, and network meta-analyses. It then deeply analyzes the misuse of the GRADE methodology in practice, including incorrect evidence grading, improper classification of evidence, misapplication in qualitative systematic reviews, inconsistencies between the documentation of the upgrading and downgrading process and results, and inappropriate use for making recommendations. Furthermore, this article comprehensively discusses the factors influencing the grading of evidence certainty in SRs/MAs of animal experiments, including the impact of bias risk, indirectness, inconsistency, imprecision, and publication bias on evidence downgrading, as well as the role of large effect sizes and cross-species consistency in evidence upgrading. Finally, in response to the issues discussed, improvement strategies are proposed, including further research and optimization of the GRADE methodology for SRs/MAs of animal experiments, the development of reporting guidelines tailored to the characteristics of SRs/MAs in animal experiment research, and enhanced professional training for researchers in the GRADE methodology. This article aims to improve the quality of evidence in SRs/MAs of animal experiments, strengthen their reliability in clinical decision-making, and promote the more efficient translation of findings from animal experiment research into clinical practice.

Objective To observe the ultrasonic manifestations of local recurrence(LR)of breast cancer after surgery.Methods Totally 82 female breast cancer patients with local hypoechoic in surgical area who underwent ultrasound follow-up were enrolled and divided into LR group(n=18)and non LR group(n=64)according to the local hypoechoic was LR or not.Clinical data and ultrasonic manifestations of primary lesion before operation and postoperative local hypoechoic were observed and compared between groups.Results Significant differences of surgical resection type,molecular subtype,status of smooth muscle actin,Calponin status,Ki-67 status,clinical staging,the maximum diameter and posterior echo of the primary lesion,as well as of involved tissue layer,location,long axis parallel to the skin or not,edge,internal echo,posterior echo,skin change and Adler blood flow grading of local hypoechoic in resection area were found between groups(all P＜0.05).Conclusion Ultrasonic manifestations of LR of breast cancer after surgery had certain characteristics.

Objective To investigate the value of afterload-related cardiac performance(ACP)in evaluating cardiac dysfunction and prognosis in patients with septic shock.Methods This prospective observational study included 45 patients with septic shock.The patients were divided into the death(n=21)or survival(n=24)group according to whether they died within 28 days.Baseline and labora-tory data were collected within 24 h.Cardiac function indices were collected and monitored at 0,6,12,18,and 24 h using the pulse index continuous cardiac output.Variables affecting the prognosis were included in the multivariate Cox proportional hazard model.Prognosis was predicted using a receiver operating characteristic(ROC)curve.Results The ACP in the death group was significantly lower than that in the survival group at 0,6,12,18,and 24 h(all P＜0.05).Logistic regression analysis showed that decreased ACP,increased acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,increased sequential organ failure assessment(SOFA)score,increased fluid balance at 24 h,increased vasoactive-inotropic score,increased central venous pressure,and decreased mean arterial pressure were risk factors for poor prognosis.The Cox proportional hazard model showed that ACP,APACHE Ⅱ score,and SOFA score were independent risk factors influencing prognosis.ROC analysis showed that the area under the curve of ACP was the largest at 24 h,which was the best time point for predicting 28-day mortality.The corresponding sensitivity,specificity,and best cutoff values were 76.20％,91.70％,and 75.16％,respectively.Conclusion ACP could be used as a valuable index to evaluate cardiac dysfunction and as an independent risk factor for poor prognosis in patients with septic shock.

Objective:To compare the efficacies of endovascular therapy (EVT) and standard medical therapy in acute ischemic stroke (AIS) patients aged ≥85 years, and analyze the independent influencing factors for poor prognosis of AIS patients after EVT.Methods:Sixty-nine AIS patients aged ≥85 years admitted to Department of Neurology, Xi'an Third Hospital from January 2018 to April 2024, including 40 accepted EVT and 28 accepted standard medicinal therapy, were enrolled. Modified Rankin scale (mRS) was used to evaluate the prognosis of the patients 90 days after onset. General data, prognosis and complications between the EVT group and standard medical therapy group were compared. General data, treatment processes and complications between patients with good prognosis and poor prognosis in the EVT group were compared. Multivariate Logistic regression was used to analyze the independent influencing factors for poor prognosis in AIS patients after EVT.Results:Compared with the standard medical therapy, the EVT group had significantly lower NIHSS score at discharge, greater improvement in NIHSS score (NIHSS score at admission-NIHSS score at discharge), lower mRS score 90 days after onset, higher good prognosis rate, lower mortality rate within 90 days of onset, and longer hospital stay ( P<0.05). In the EVT group, 11 patients (27.5%) had good prognosis and 29 patients (72.5%) had poor prognosis 90 days after onset. Compared with the good prognosis group, the poor prognosis group had significantly higher blood glucose level and lower Alberta Stroke Program Early CT Score (ASPECT) on admission ( P<0.05). Multivariate Logistic regression analysis showed that blood glucose on admission ( OR=2.363, 95% CI: 1.134-4.928, P=0.022) and ASPECT score on admission ( OR=0.273, 95% CI: 0.088-0.854, P=0.026) were independent influencing factors for poor prognosis in AIS patients after EVT. Conclusion:AIS patients aged ≥85 years received EVT have better prognosis compared with those accepted standard medical therapy; these patients with high glucose level and low ASPECT score on admission have poor prognosis.

Objective:To investigate the impact of flow diverter (FD) malapposition at the aneurysm neck on clinical outcomes and complications of intracranial aneurysms, and identify the influencing factors for intraoperative FD malapposition.Methods:A retrospective study was performed; 153 patients with unruptured saccular aneurysms at the C4-C7 segments of the internal carotid artery accepted single FD implantation at Department of Interventional Radiology, First Affiliated Hospital of Zhengzhou University from June 2022 to March 2024 were chosen. Intraoperative high-resolution C-arm CT was utilized to assess FD apposition at the aneurysm neck. (1) Based on FD apposition at the aneurysm neck as shown, these 153 patients were divided into a malapposition group ( n=23, including 16 patients with malapposition being identified as residual malapposition after intraoperative corrective measures such as microwire massage and 7 patients with malapposition being newly detected in this study) and a complete apposition group ( n=130). Perioperative and follow-up complications were recorded. Clinical outcomes were assessed using modified Rankin Scale (mRS) at the final follow-up (mRS score of 0-2 as favorable outcome), and angiographic outcomes were evaluated by DSA at the final follow-up. Differences in clinical and angiographic outcomes and complication rate were compared between the malapposition group and complete apposition group. (2) Based on FD apposition at the aneurysm neck as shown, these 153 patients were divided into an intraoperative malapposition group ( n=74, including 67 patients with malapposition being detected during surgery and 7 patients with malapposition being newly detected in this study) and an intraoperative complete apposition group ( n=79). Univariate analysis was performed to compare the clinical variables between the intraoperative malapposition group and intraoperative complete apposition group; multivariate Logistic regression was further employed to identify the independent influencing factors for FD malapposition at the aneurysm neck. Results:(1) Four patients (all from the malapposition group) developed perioperative acute in-stent thrombosis. Nine patients experienced ischemic or hemorrhagic stroke during the follow-up, including 6 from the malapposition group and 3 from the complete apposition group; the complication rate in the malapposition group (6/23, 26.1%) was significantly higher than that in the complete apposition group (3/130, 2.3%) during the follow-up ( P<0.05). At the final follow-up, 2 patients (both from the malapposition group) had poor clinical outcome, while the remaining 151 patients had favorable outcome. Proportion of patients with favorable outcome between the two groups was statistically different (91.3%[21/23] vs. 100.0%[130/130], P<0.05). Delayed occlusion was detected in 46 patients (12 from the malapposition group and 34 from the complete apposition group) at the final angiographic follow-up. FD restenosis/re-occlusion was noted in 10 patients, including 6 from the malapposition group and 4 from the complete apposition group. Significant difference in delayed occlusion rate (52.2%[12/23] vs. 26.2%[34/130]) and long-term in-stent stenosis/occlusion rate (26.1%[6/23] vs. 3.1%[4/130]) was observed between the two groups ( P<0.05). (2) Significant difference in aneurysm neck diameter, FD angulation, parent artery stenosis, parent artery diameter ratio>1.2, and presence of branching vessels at the FD implantation site was noted between the intraoperative complete apposition group and intraoperative malapposition group ( P<0.05). Multivariate Logistic regression indicated that aneurysm neck diameter ( OR=1.431, 95% CI: 1.096-1.868, P=0.008), parent artery diameter ratio>1.2 ( OR=2.199, 95% CI: 1.083-4.463, P=0.029), and FD angulation ( OR=1.019, 95% CI: 1.002-1.036, P=0.027) were independent influencing factors for FD malapposition at the aneurysm neck. Conclusion:In FD implantation for intracranial aneurysms, FD malapposition at the aneurysm neck adversely affects delayed occlusion rate and complication rate; aneurysms with wider aneurysm neck diameter, parent artery diameter ratio>1.2, and greater FD angulation are trend to have FD malapposition at the aneurysm neck.

Objective To investigate the value of afterload-related cardiac performance(ACP)in evaluating cardiac dysfunction and prognosis in patients with septic shock.Methods This prospective observational study included 45 patients with septic shock.The patients were divided into the death(n=21)or survival(n=24)group according to whether they died within 28 days.Baseline and labora-tory data were collected within 24 h.Cardiac function indices were collected and monitored at 0,6,12,18,and 24 h using the pulse index continuous cardiac output.Variables affecting the prognosis were included in the multivariate Cox proportional hazard model.Prognosis was predicted using a receiver operating characteristic(ROC)curve.Results The ACP in the death group was significantly lower than that in the survival group at 0,6,12,18,and 24 h(all P＜0.05).Logistic regression analysis showed that decreased ACP,increased acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,increased sequential organ failure assessment(SOFA)score,increased fluid balance at 24 h,increased vasoactive-inotropic score,increased central venous pressure,and decreased mean arterial pressure were risk factors for poor prognosis.The Cox proportional hazard model showed that ACP,APACHE Ⅱ score,and SOFA score were independent risk factors influencing prognosis.ROC analysis showed that the area under the curve of ACP was the largest at 24 h,which was the best time point for predicting 28-day mortality.The corresponding sensitivity,specificity,and best cutoff values were 76.20％,91.70％,and 75.16％,respectively.Conclusion ACP could be used as a valuable index to evaluate cardiac dysfunction and as an independent risk factor for poor prognosis in patients with septic shock.

Objective To explore the diagnostic value of lung ultrasound(LUS)score,computed tomo-graphy(CT)combined with serum CXC chemokine ligand 8(CXCL8)and soluble co stimulatory molecule B7-H3(sB7-H3)for severe Mycoplasma pneumoniae pneumonia(MPP)in children.Methods From April 2022 to April 2024,a total of 210 children with MPP who visited Maternal and Child Health Hospital of Zhan-gqiu District were included as research subjects,and they were separated into severe MPP group and non se-vere MPP group according to the severity of the disease.Enzyme-linked immunosorbent assay was applied to detect serum levels of CXCL8 and sB7-H3.LUS score and CT diagnosis were conducted.The value of LUS score,CT and serum CXCL8,sB7-H3 in the diagnosis of severe MPP was analyzed using the receiver operating characteristic curve.Results There were no statistically significant differences in height,age,fever days,weight,hospital stay,and gender between the severe MPP group and the non severe MPP group(P>0.05).Compared with the non severe MPP group,the serum levels of CXCL8,sB7-H3,and LUS score were all in-creased in the severe MPP group(P<0.05).The area under the curve(AUC)of LUS score,serum CXCL8,and sB7-H3 for diagnosing severe MPP was 0.865,0.785,and 0.750,respectively.The CT diagnosis results showed 35 false positives and 22 false negatives,with Kappa value of 0.459 compared to clinical examination results(P<0.05).The diagnostic results of LUS score,CT combined with serum CXCL8 and sB7-H3 showed 3 false positives and 15 false negatives,with Kappa vale of 0.828 compared to clinical examination results(P<0.05).The diagnosis of LUS score,CT combined with serum CXCL8,and sB7-H3 had higher specificity and accuracy than those of single detection of the four(P<0.05),and the sensitivity of the combined diagno-sis of the four was increased(P<0.05)and the missed diagnosis rate was reduced compared to the single de-tection of CXCL8 and sB7-H3(P<0.05).Conclusion The serum levels of CXCL8,sB7-H3,and LUS score in children with MPP increase.The specificity and accuracy of LUS score,CT combined with serum CXCL8,sB7-H3 in diagnosing severe MPP increase.

Objective: To analyze the correlation between serum homocysteine (Hcy) and both folic acid (FA) and sperm DNA fragmentation index (DFI), so as to provide the evidence for male fertility assessment. Methods: Males who visited and measured the serum Hcy in the Reproductive Medicine Center of Huzhou Maternal and Child Health Care Hospital from September 2022 to September 2023 were selected as the study subjects. Sperm quality parameters and sperm DFI were analyzed by collecting sperm. Hcy and FA were measured by collecting venous blood. Participants were stratified into a high Hcy group (Hcy≥15.0 μmol/L) and a normal group (Hcy<15.0 μmol/L). The correlations between serum Hcy and FA and sperm DFI were evaluated using linear regression models. Results: A total of 173 participants were enrolled, including 39 in the high Hcy group and 134 in the normal group. The sperm concentration in the high Hcy group was significantly lower than that in the normal group [(91.77±61.11)×106/mL vs. (144.21±106.82)×106/mL, P<0.05]. No statistically significant differences were observed in semen volume, sperm motility, curvilinear velocity, straight-line velocity, average path velocity, or sperm morphology normal rate (all P>0.05). The FA level in the high Hcy group was lower than that in the normal group [(4.44±1.79) nmol/L vs. (7.64±3.68) nmol/L, P<0.05]. The sperm DFI in the high Hcy group was higher than that in the normal group [(19.21±8.85)% vs. (13.07±6.43)%, P<0.05]. Serum Hcy level showed a negative correlation with FA level (r=-0.369, P<0.05) and a positive correlation with sperm DFI (r=0.351, P<0.05). Conclusion Serum Hcy level is associated with sperm concentration, FA and sperm DFI, suggesting that serum Hcy may affect sperm quality.