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.

To investigate the association between neutrophil to lymphocyte ratio (NLR) andestimated glomerular filtration rate (eGFR) in patients with diabetes using large-scale data.

BACKGROUND:Acute myocardial infarction can cause cardiac remodeling and heart failure,as well as skeletal myopathy,affecting patients'quality of life.Exercise therapy is an important rehabilitation method for patients with heart failure;however,the optimal exercise prescription has not been clarified. OBJECTIVE:To compare the effects of different exercise modes(aerobic exercise,resistance exercise)on skeletal muscle remodeling in rats with acute myocardial infarction induced heart failure and to explore the possible mechanism,so as to provide a basis for optimizing the exercise rehabilitation program. METHODS:Forty-eight Sprague-Dawley rats were randomly divided into sham operation group,myocardial infarction group,aerobic exercise group and resistance exercise group.Coronary artery ligation was used to create model of heart failure.After 3 months,animals in the aerobic exercise group and resistance exercise group underwent 12 weeks of corresponding exercise mode interventions,while those in the sham operation group and myocardial infarction group were kept quietly in mouse cages.After the experiment,maximal running speed and maximal weight-bearing load were measured by graded treadmill exercise test and ladder-climbing test respectively,and heart structure and function were evaluated by echocardiography.The heart was isolated,and hematoxylin-eosin staining and Sirius red staining were performed to detect cardiac remodeling.For the gstrocnemius muscle,ATPase staining was performed to observe changes in muscle fiber type and cell cross-sectional area,dihydroethidium method was used to evaluate reactive oxygen species levels,enzyme-linked immunosorbent method was used to determine malondialdehyde content and antioxidant enzyme activity,western blot was used to determine the expression of ubiquitin-proteasome system proteins,and the number of activated satellite cells(Pax7+/MyoD+)were detected by double immunofluorescence staining. RESULTS AND CONCLUSION:(1)Exercise performance:Compared with the sham operation group,maximal running speed and maximal weight-bearing load in the myocardial infarction group decreased(P<0.05);compared with the myocardial infarction group,the maximal running speed of the aerobic exercise group and the maximal weight-bearing load of the resistance exercise group increased(P<0.05).(2)Cardiac remodeling:Compared with the sham operation group,infarction area,myocardial cell cross-sectional area,and collagen content in the myocardial infarction group increased(P<0.05),while leftventricular ejection fraction and shortening fraction decreased(P<0.05);compared with the myocardial infarction group,there was no statistical difference in the above parameters in both aerobic exercise resistance exercise groups(P>0.05).(3)Skeletal muscle remodeling:Compared with the sham operation group,gastrocnemius muscle mass,gastrocnemius muscle mass index,cell cross-sectional area,superoxide dismutase activity,glutathione peroxidase activity,and the number of activated satellite cells decreased in myocardial infarction group(P<0.05),while reactive oxygen species content,malondialdehyde content,and the protein expression of ubiquitin,MuRF1 and MAFbx increased(P<0.05);compared with the myocardial infarction group,gastrocnemius muscle mass index,superoxide dismutase activity,the number of activated satellite cells increased in both aerobic exercise and resistance exercise groups(P<0.05),while reactive oxygen species content and the protein expression of ubiquitin,MuRF1,and MAFbx decreased(P<0.05);compared with the aerobic exercise group,gastrocnemius muscle mass,gastrocnemius muscle mass index,cell cross-sectional area,reactive oxygen species content,malondialdehyde content,the number of activated satellite cells increased in resistance exercise group(P<0.05),while superoxide dismutase activity,glutathione peroxidase activity down-regulated(P<0.05).To conclude,aerobic exercise and resistance exercise can both improve exercise performance of rats with heart failure,and the mechanism is related to reducing oxidative stress,inhibiting ubiquitin-proteasome system activity and activating satellite cells to improve skeletal muscle remodeling.Aerobic exercise has a better effect on improving skeletal muscle oxidative stress,while resistance exercise has a more significant effect on promoting skeletal muscle regeneration.

Receptor-interacting serine/threonine-protein kinase 1 (RIPK1) functions as a key regulator in inflammation and cell death and is involved in mediating a variety of inflammatory or degenerative diseases. A number of allosteric RIPK1 inhibitors (RIPK1i) have been developed, and some of them have already advanced into clinical evaluation. Recently, selective RIPK1i that interact with both the allosteric pocket and the ATP-binding site of RIPK1 have started to emerge. Here, we report the rational development of a new series of type-II RIPK1i based on the rediscovery of a reported but mechanistically atypical RIPK3i. We also describe the structure-guided lead optimization of a potent, selective, and orally bioavailable RIPK1i, 62, which exhibits extraordinary efficacies in mouse models of acute or chronic inflammatory diseases. Collectively, 62 provides a useful tool for evaluating RIPK1 in animal disease models and a promising lead for further drug development.

Objective:This study was designed to explore the distribution pattern of TCM syndrome types in patients with Chronic Kidney Disease (CKD) stage 3-5 without alternative treatment after objective collection of TCM quad-diagnostic instruments.Methods:The four-diagnostic instruments of Chinese medicine were used to collect the four-diagnostic information of patients with stage CKD 3-5 non-alternative treatment for syndrome determination, and the correlation between TCM syndrome and basic disease characteristics of patients was analyzed.Results:The distribution of TCM syndrome types in 464 patients with CKD 3-5 stage non-substitution therapy was based on deficiency syndrome, and had both standard and solid syndrome. Qi-deficiency syndrome was the most common type, accounting for 24.6% (114/464), followed by kidney-yang deficiency syndrome, heart-qi deficiency syndrome, kidney-yin deficiency syndrome, heart-blood deficiency syndrome, spleen-yang deficiency syndrome and lung-yin deficiency syndrome. The positivism type of this deficiency is blood stasis, dampness-heat, moisture, turbidity and turbidity toxicity. There was no significant difference in gender and age distribution among patients with different CKD stages ( P>0.05), but the proportion of deficiency syndrome gradually increased with the increase of age. There were differences in the distribution of primary deficiency syndrome in different CKD stages ( χ2=57.48, P<0.001), but no difference in the distribution of primary deficiency syndrome ( χ2=2.59, P=0.957). Conclusions:According to the four diagnostic instrument of traditional Chinese medicine, the distribution of TCM syndrome types in patients with stage CKD3-5 non-alternative treatment is based on deficiency syndrome, combined with deficiency of primary and solid syndrome. The syndrome types in CKD3 stage were mainly qi deficiency and kidney qi deficiency, while the TCM syndrome types in CKD stage 4 were qi deficiency and kidney Yang deficiency. With the progression of the disease, the TCM syndromes of stage 5 CKD were mainly heart-qi deficiency and kidney-yang deficiency.

Objective:To explore the mechanism of Mito-TEMPO inhibiting the activation of BV2 microglia induced by lead(Pb)exposure.Methods:Mouse microglia BV2 were cultured in vitro.The effects of different concentrations of lead exposure on the viability of BV2 cells were determined by MTT colorimetric method,and a model of lead expo-sure was developed and intervened with Mito-TEMPO antioxidant.Immunofluorescence staining was used to detect the expression of M1 activation marker CD86 protein.Enzyme-linked immunosorbent assay was used to detect the expression of pro-inflammatory factors interleukin-1 β(IL-1β),tumor necrosis factor α(TNF-α),interleukin-6(IL-6).Mito-chondrial superoxide indicator(MitoSOX)staining was used to detect the level of mitochondrial oxidative stress.JC-1 staining was used to detect mitochondrial membrane potential.The respiratory ability of mitochondria was detected by cell energy metabolism analysis system(O2k).Results:Compared with the control group,the expression of CD86 protein,the levels of pro-inflammatory cytokines IL-1β,TNF-α and IL-6 in BV2 cells increased significantly,the level of oxidative stress in mitochondria increased significantly,and the mitochondrial membrane potential and respiratory ability decreased significantly in lead exposed group.Mito-TEMPO treatment significantly reduced the oxidative stress and functional damage of mitochondria in BV2 cells,and significantly inhibited the M1 activation level of BV2 cells.Conclusion:The results show that Mito-TEMPO treatment can reverse the M1 activation of BV2 cells induced by lead exposure,and the specific mechanism may be related to the reduction of mitochondrial oxidative stress and dysfunction by Mito-TEMPO.

AIM To investigate the variation rules of main secondary metabolites in Hedysari Radix before and after rubbing strip.METHODS UPLC-MS/MS was adopted in the content determination of formononetin,ononin,calycosin,calycosin-7-glucoside,medicarpin,genistein,luteolin,liquiritigenin,isoliquiritigenin,vanillic acid,ferulic acid,γ-aminobutyric acid,adenosine and betaine,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were used for chemical pattern recognition to explore differential components.RESULTS After rubbing strip,formononetin,calycosin,liquiritigenin and γ-aminobutynic acid demonstrated increased contents,along with decreased contents of ononin,calycosin-7-glucoside and vanillic acid.The samples with and without rubbing strip were clustered into two types,calycosin-7-glucoside,formononetin,γ-aminobutynic acid,vanillic acid,calycosin-7-glucoside and formononetin were differential components.CONCLUSION This experiment clarifies the differences of chemical constituents in Hedysari Radix before and after rubbing strip,which can provide a reference for the research on rubbing strip mechanism of other medicinal materials.

Objective:To explore the values of single and combined detection of mammography,ultrasound Doppler and serum markers of tumor included serum prostate specific antigen(PSA),serum carbohydrate antigen 15-3(CA153),mucin 1(MUC1)and human growth differentiation factor 3(GDF3)in diagnosing early breast cancer.Methods:A total of 96 patients with breast cancer,who admitted to Tangshan People's Hospital from January 2018 to December 2021 and were confirmed by pathological examination,were selected as breast cancer group.At the same time,70 patients with benign breast diseases who received diagnosis and treatment in our hospital were selected as benign lesions group.In addition,50 normal people who were confirmed as health by physical examination in our hospital were selected as research subjects of healthy control group.The postoperative pathological examination was used as the gold standard to compare the diagnostic values of single mammography,ultrasonic Doppler examination,serum PSA,CA153,MUC1,GDF3 and the combined examination of them for breast cancer.Results:In the breast cancer group,78 cases of the 96 patients with breast cancer were diagnosed as malignant tumor by ultrasound on breast,with a positive detection rate of 81.3%,and 80 cases of them were diagnosed as malignant tumor by mammography X-ray examination,with a positive detection rate of 83.1%.The levels of serum PSA,CA153,MUC1 and GDF3 of breast cancer group were respectively higher than those of the benign lesion group and healthy control group,and the differences were statistically significant(t=8.783,10.361,11.258,18.965,9.564,12.658,12.688,20.163,P<0.05).Using breast cancer as the dependent variable,and using serum PSA,CA153,MUC1 and GDF3 as independent variable to perform Logistic regression analysis.The results of Logistic regression analysis indicated that serum PSA,CA153,MUC1 and GDF3 were important risk factors of breast cancer(OR value =1.165,1.168,1.472,1.248,P<0.05).The results of receiver operating characteristic(ROC)curve(95%CI),sensitivity and specificity of single application of each indicator of ultrasound on breast,mammography,serum PSA,CA153,MUC1 and GDF3 were respectively[0.723(0.595-0.851),82.56%and 67.32%],[0.761(0.636-0.886),85.79%and 65.36%],[0.833(0.726-0.941),81.48%and 85.73%],[0.837(0.738-0.926),61.25%and 70.17%],[0.768(0.648-0.889),71.49%and 80.87%],[0.613(0.469-0.758),52.94%and 50.57%].However,the AUC(95%CI),sensitivity and specificity of the combined application of 6 items were respectively 0.958(0.905-0.999),96.37%and 84.83%,which had higher diagnostic efficiency.Conclusion:The combined detection performance of mammography,ultrasound Doppler and serum PSA,CA153,MUC1 and GDF3 is higher than that of single each detection,which is helpful to conduct early identification and diagnosis for breast cancer.

ObjectiveTo explore the protective effect and mechanism of Yangxin Dingji capsules on isoproterenol （ISO）-induced ventricular arrhythmia in SD rat cardiomyocytes based on the transforming growth factor-β activated kinase 1 （TAK1）-mitogen-activated protein kinase kinase 3 （MKK3）-p38 mitogen-activated protein kinase （p38 MAPK） signaling pathway. MethodFifty male SPF-grade SD rats were randomly divided into a normal group， a model group， a propranolol group， a low-dose Chinese medicine group， and a high-dose Chinese medicine group. The ventricular arrhythmia model was constructed using the ISO "6+1" method. The propranolol group received propranolol at 0.015 g·kg^-1·d^-1. The Chinese medicine groups received Yangxin Dingji capsules at doses of 0.5、 2 g·kg^-1·d^-1， respectively. The normal and model groups were given an equal volume of 0.9% NaCl solution. Electrocardiogram （ECG） changes in SD rats were recorded using the BL-420F biological function experimental system. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in the heart. Serum levels of cardiac troponin I （cTnI）， creatine kinase-MB （CK-MB）， interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， and transforming growth factor-β₁ （TGF-β₁） were measured using enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of IL-1β and TNF-α was assessed using real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. Reactive oxygen species （ROS） expression was detected using immunofluorescence. Protein expression levels of TAK1， phosphorylated TAK1 （p-TAK1）， MKK3， phosphorylated MKK3 （p-MKK3）， p38 MAPK， phosphorylated p38 MAPK （p-p38 MAPK）， nuclear factor-κB （NF-κB）， phosphorylated NF-κB （p-NF-κB）， IL-1β， and TNF-α were measured using Western blot or immunohistochemistry. ResultCompared with normal group， the model group showed significant ventricular arrhythmia in ECG， with an increased arrhythmia score （P<0.01）. Pathological damage to myocardial tissue was evident， and serum levels of cTnI， CK-MB， IL-1β， TNF-α， and TGF-β₁ were elevated （P<0.01）. The mRNA and protein expression of IL-1β and TNF-α in myocardial tissue was also increased （P<0.01）. ROS level and protein expression of p-TAK1， p-MKK3， p-p38 MAPK， and p-NF-κB were elevated in myocardial tissue （P<0.01）. In the propranolol and Chinese medicine groups， the incidence of sustained ventricular tachycardia （SVT） and arrhythmia scores were significantly reduced compared to model group （P<0.05， P<0.01）. Pathological damage to cardiomyocytes was alleviated， and levels of myocardial injury markers and inflammatory factors in serum and myocardial tissue were decreased. The ROS level in myocardial tissue was also reduced （P<0.01）， with a noticeable reduction in related molecules in the p38 MAPK pathway （P<0.05， P<0.01）. ConclusionThe expression of p38 MAPK pathway molecules was up-regulated in myocardial tissue of ISO-induced ventricular arrhythmia rats. Yangxin Dingji capsules may inhibit cardiac inflammation damage by regulating the expression of related molecules in the p38 MAPK pathway， thereby exerting a protective effect on myocardial cells， with TAK1 being a potential target.

Objective To explore the mechanism of ginseng in the treatment of periodontitis based on network phar-macology and molecular docking technology.Methods Potential targets of ginseng and periodontitis were obtained through various databases.The intersection targets of ginseng and periodontitis were obtained by using VENNY,the pro-tein-protein interaction network relationship diagram was formed on the STRING platform,the core target diagram was formed by Cytoscape software,and the ginseng-active ingredient-target network diagram was constructed.The selected targets were screened for gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway en-richment analysis.The core targets of ginseng's active in-gredients in treating periodontitis were analyzed by mo-lecular docking technique.Results The 22 ginseng's active ingredients,591 potential targets of ginseng's ac-tive ingredients,2 249 periodontitis gene targets,and 145 ginseng-periodontitis intersection targets were analyzed.Ginseng had strong binding activity on core targets such as vas-cular endothelial growth factor A and epidermal growth factor receptor,as well as hypoxia induced-factor 1(HIF-1)sig-naling pathway and phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt)signaling pathway.Conclusion Gin-seng and its active components can regulate several signaling pathways such as HIF-1 and PI3K-Akt,thereby indicating that ginseng may play a role in treating periodontitis through multiple pathways.