OBJECTIVE To investigate the mechanism by which Naozhenning granules （NZN） improve learning and memory impairment in a rat model of multiple cerebral concussion （MCC）. METHODS The MCC rat model was established using the closed controlled cortical impact method. The experiment was set up with a blank group （normal saline）， a model group （normal saline）， a piracetam group （positive control group， 0.324 g/kg）， and high-， medium-， and low-dose NZN groups （5.4， 2.7， 1.35 g/kg）， with 11 rats in each group. Drugs or normal saline were administered by gavage once daily for 28 consecutive days. General condition and body weight were monitored throughout the experiment. The sucrose preference rate and novel object recognition index were measured； Evans blue （EB） extravasation in the cerebral cortex was detected； pathological changes of cortical neurons were observed； the levels of B-cell lymphoma-2 （Bcl-2）， Bcl-2-associated X protein （Bax）， interleukin-6 （IL-6）， IL-10， and tumor necrosis factor-α （TNF-α） in the cerebral cortex were determined； and the phosphorylation levels of AMP-activated protein kinase （AMPK）， glycogen synthase kinase 3β （GSK3β）， and Tau protein were detected. RESULTS Compared with the blank group， the model group showed poor mental state， sluggish response to external stimuli， reduced food and water intake， decreased limb flexibility， and disheveled fur. Body weight， sucrose preference rate， and novel object recognition index were significantly decreased （ P ＜0.05）； EB extravasation in the cerebral cortex was significantly increased （ P ＜0.05）， with severe neuronal damage. The positive area ratio of Bax protein， IL-6 and TNF-α levels， and Tau protein phosphorylation level were all significantly increased （ P ＜0.05）， whereas the positive area ratio of Bcl-2 protein， IL-10 level， and AMPK and GSK3β protein phosphorylation levels were significantly decreased （ P ＜0.05）. Compared with the model group， all NZN dose groups showed improvements in general condition and pathological damage， with quantitative indices partially restored， and the differences in quantitative indices in high-dose NZN group were statistically significant （ P ＜0.05）. CONCLUSIONS NZN can effectively improve learning and memory impairment in MCC model rats. The mechanism may be related to activating the AMPK/GSK3β pathway， inhibiting inflammatory response， reducing Tau protein phosphorylation level， and then repairing the neuronal injury.

This study elucidated the structure-activity relationships (SAR) of three distinct opioid ligand series at the μ-opioid receptor (μ-OR) and investigated the ligand-dependent role of arrestin in downstream signaling. Radioligand binding assays across 13 compounds revealed that high-affinity μ-OR binding is not restricted to a single chemical scaffold. For fentanyl analogs, SAR analysis identified four key structural determinants: the N-phenethyl group (R1) is essential for μ-OR binding, the piperidine 4-position (R2) is sterically constrained and tolerates only small substituents such as carbomethoxy, the acyl side chain (R3) tolerates diverse chemical modifications including alkyl, alkoxy, and heteroaryl groups without substantial loss of affinity, and para-substitution on the phenethyl ring (R4) with electron-withdrawing groups like fluorine is well-tolerated. Analysis of non-fentanyl scaffolds (2-Methyl AP-237, W-15, and brorphine) demonstrated that specific side-chain functionalization, rather than core scaffold architecture, primarily determines binding potency. Comparison of classical opioids further revealed that structural flexibility, exemplified by methadone, can confer superior binding affinity compared to the rigid morphine scaffold. Consistent with this structural diversity, the contribution of arrestins to ERK activation varied substantially across compounds, suggesting that μ-OR-biased signaling is not dictated exclusively by intrinsic receptor properties but emerges from the interplay between receptor conformation and ligandspecific structural features. Collectively, our results provide a molecular framework for understanding opioid pharmacology with important implications for rational drug design aimed at minimizing adverse effects while maintaining analgesic efficacy.

ObjectiveTo construct a prediction model for high responsiveness to short-term aerobic training in patients with Parkinson's disease (PD) based on dual-task gait assessment, and to provide evidence for identifying individuals who are most likely to benefit from such interventions. MethodsA total of 42 patients with PD who visited the outpatient clinic of Tianjin Huanhu Hospital between August 29, 2023 and December 19, 2024 were enrolled. All participants completed a 4-week home-based aerobic cycling training program and underwent dual-task gait assessments pre- and post-training. Patients were classified into high-response group (n = 12) and low-response group (n = 30) according to whether the improvement in gait speed reached the minimal clinically important difference of 0.1 m/s. The clinical validity of this grouping was verified by comparing differences in cardiopulmonary exercise testing parameters and dual-task gait indicators between the two groups pre- and post-training. Based on baseline characteristics, Firth's penalized maximum likelihood logistic regression was applied to construct a predictive model for high responsiveness, and the discriminative performance of the model was assessed preliminarily. ResultsThe high-response group showed significantly greater improvement in gait speed, step length, cadence and the anaerobic threshold to maximal oxygen uptake ratio after training compared with the low-response group (P < 0.05), confirming the clinical validity of the grouping. The baseline anaerobic threshold to maximal oxygen uptake ratio (OR = 3.348), body mass index (OR = 2.229) and age (OR = 0.428) demonstrated certain discriminative ability in the present sample. The area under the receiver operating characteristic curve was 0.831, with an overall prediction accuracy of 81.0%, specificity of 83.3% and sensitivity of 75.0%. Robustness of the model was confirmed via internal validation using the Bootstrap method, which suggested that the results were informative, but further validation was still required. ConclusionThe anaerobic threshold/maximal oxygen uptake ratio, body mass index and age may predict the response to short-term aerobic training in patients with PD.

ObjectiveTo explore the ameliorative effect of the endoplasmic reticulum stress inhibitor 4-phenylbutyric acid （4-PBA） on bile acid metabolism disorders induced by bacterial lipopolysaccharide （LPS） in late-pregnant mice， and to clarify the regulatory mechanism of endoplasmic reticulum stress （ERS） in intrahepatic cholestasis of pregnancy （ICP）. MethodsTwenty-four pregnant mice were divided into control group， LPS group， 4-PBA group， and LPS+4-PBA group. On the 16th day of pregnancy， the LPS group received intraperitoneal injection of LPS （200 μg/kg） and the 4-PBA group received injection of 4-PBA （150 mg/kg）. In the LPS+4-PBA group， 4-PBA was intraperitoneally injected first， followed by LPS administration 1 h later. Serum total bile acid （TBA）， liver injury markers， bile acid profiles， and the expression of key genes and proteins were detected 6 h after LPS treatment. ResultsLPS exposure increased serum TBA levels in pregnant mice（P<0.05）， activated ERS markers and inflammatory factors， and downregulated bile acid transporters and the expression of the rate limiting enzyme CYP7A1（all P<0.05）. 4-PBA intervention effectively inhibited ERS， reduced TBA levels（P<0.05）， and upregulated Ntcp， Bsep， Mdr3， and Mrp3 gene expression（all P<0.05）. ConclusionERS plays an important role in LPS interference with the homeostasis of bile acid metabolism during pregnancy. Inhibiting ERS can improve bile stasis by regulating bile acid transporters， providing a theoretical basis for targeted therapy of ICP.

Objective To investigate the correlation between the triglyceride-glucose(TyG)index,its derivatives in combination with obesity indicators and the risk of albuminuria in patients with type 2diabetes mellitus(T2DM).Methods T2DM inpatients who were hospitalized in the Department of Endocrinology from January 2020 to March 2023 were consecutively collected as study subjects,and divided into the group with no albuminuria(n=246)and the group with albuminuria(n=221).Spearman correlation analysis was used to analyze the correlation between TyG index,its derivative index and clinical indicators,and multivariate Logistic regression was used to analyze the correlation between TyG index,its derivative indexes and albuminuria.Results The TyG index and its derived indices[TyG-body mass index(TyG-BMI),TyG-waist-to-hip ratio(TyG-WHR),TyG-waist-to-height ratio(TyG-WtHR)]were significantly higher in the albuminuric group of patients with T2DM than in the group without albuminuria(all P＜0.001).The inci-dence of albuminuria gradually increased with increasing tertile groupings of all four indicators(all P＜0.001).In multivariate Logistic regression analysis,after correcting for screened risk factors,the TyG index(OR=1.50,95％CI:1.07-2.08,P=0.019),TyG-BMI(OR=1.01,95％CI:1.00-1.01,P=0.009),TyG-WHR(OR=1.45,95％CI:1.12-1.88,P=0.005),and TyG-WtHR(OR=1.39,95％CI:1.00-1.91,P=0.048)were the independent risk factors for albuminuria.For categorical variables,using the 1st quartile group(Q1)as a control group,the 3rd quartile group(Q3)of all four indices were all risk factors for albuminuria,whereas the Q3group of TyG-WHR was more significantly associated with albuminuria compared to the TyG index alone and the other de-rived indices(OR=2.55,95％CI:1.44-4.51,P=0.002).Conclusion In patients with T2DM,both TyG index and its derivatives combined with obesity indicators were significantly associated with albuminuria,and in patients with higher index levels,the risk of albu-minuria was higher in patients with high levels of TyG-WHR compared with single TyG index and other derivatives.Therefore,TyG-WHR may be a novel biological indicator to predict the development of DKD in T2DM patients.

Objective:To develop a method of employing flow cytometry to directly detect the γ-H2AX expression levels in peripheral blood lymphocytes of mice through fixation and lysis and to evaluate the feasibility of applying this method to research on the radiation-related biological effects and the efficacy evaluation of radioprotective drugs.Methods:A total of 41 male C57BL/6J mice were used. First, 21 mice were randomly divided into 7 groups according to different radiation doses (0, 1, 2, 4, 6, 8, and 10 Gy) with 3 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation and immediately fixed with formaldehyde. Red blood cells (RBC) were lysed with Triton X-100, and γ-H2AX was labeled with specific antibodies. DRAQ5 dye was used to further exclude debris and anucleate cells. The mean fluorescence intensity of γ-H2AX in lymphocyte populations was directly analyzed by flow cytometry through forward and side scatter, and dose-effect curves after irradiation were established. Then, the other 20 mice were divided into radiation alone groups and radiation combined with WR-2721 administration groups at 4 and 6 Gy, respectively, with 5 mice in each group. Blood samples were collected from the tail vein of mice at 1, 4, 8, and 24 h after irradiation to detect the average fluorescence intensity of γ-H2AX in lymphocytes, which was used to evaluate the degree of DNA damage in mice and the therapeutic effect of WR-2721.Results:The expression of γ-H2AX in peripheral blood lymphocytes of mice significantly increased with the increase of radiation doses, and reached a peak at 1-2 h and then decreased. The dose-effect relationship was significant ( R2 = 0.9914). At 24 h after 4 and 6 Gy irradiation, compared with the radiation alone groups, the average fluorescence intensity of γ-H2AX in the radiation combined with WR-2721 administration groups was lower (144.8 ± 8.0 and 109.5 ± 9.7, vs. 178.0 ± 18.5 and 136.6 ± 5.4), with statistically significant difference ( t = 3.78, 5.48, P < 0.05). The average fluorescence intensity of γ-H2AX at 24 h after irradiation was consistent with the lowest values of the three blood cell lines at 7 or 14 d after irradiation. Conclusions:The application of flow cytometry with a fixation/dissolution protocol to directly detect the mean fluorescence intensity of γ-H2AX in peripheral blood lymphocytes of mice has significant application value in radiation biology effect research, radiation protection drug screening, and efficacy evaluation.

Objective To investigate the diagnostic efficacy of fat fraction(FF)and T2* values of infrapatellar fat pad(IPFP)based on MR mDixon-Quant technique for knee osteoarthritis(KOA),and to analyze their related influencing factors.Methods A total of 178 knee joints from 114 volunteers were included and underwent knee joints X-ray examination and MR mDixon-Quant sequence scanning.The correlation between FF and T2* values of IPFP with gender,age,body mass index(BMI),and K-L grade was analyzed by Spearman's test and multiple linear regression.Receiver operating characteristic(ROC)curves were plotted to assess the diag-nostic efficacy of FF and T2* values for KOA.Results FF value of IPFP was negatively correlated with age,BMI,and K-L grade(r=-0.470,-0.276,-0.679,P＜0.001),and T2* value was also negatively correlated with age,BMI,and K-L grade(r=-0.291,-0.173,-0.505,P＜0.001).Multiple linear regression analysis results showed that only K-L grade had a statistically significant correlation with FF and T2* values(P＜0.001).ROC curve analysis indicated that the area under the curve(AUC)for diagnosing KOA using the FF and T2* values of IPFP were 0.859 and 0.767,respectively,with sensitivity of 0.878 and 0.841,and specificity of 0.719 and 0.573,respectively.The combined diagnosis had an AUC of 0.855,a sensitivity of 0.625,and a specificity of 0.963,improving the diagnostic specificity for KOA.Conclusion FF and T2* values of IPFP can serve as objective imaging indicators for KOA assessment,and their reduction is correlated with the progress of age,BMI,and K-L grade,of which the K-L grade is an independent influencing factor.The combination of FF and T2 can improve the diagnostic specificity of KOA,and provide a quantitative basis for early KOA detection and severity stratification.

BACKGROUND:Steroid-induced osteonecrosis of the femoral head is closely related to ferroptosis in osteoblasts.Deer antler peptide can promote the survival and functional establishment of osteoclasts by inhibiting ferroptosis in osteoblasts,and has the potential to treat steroid-induced osteonecrosis of the femoral head,but its regulatory mechanism of ferroptosis in osteoblasts has not yet been clarified.OBJECTIVE:To investigate the mechanism by which deer antler peptide inhibits dexamethasone-induced ferroptosis in osteoblasts.METHODS:(1)Different concentration gradients of antler peptide and dexamethasone were used to intervene in MC3T3-E1 14 cells,and the cell activity was detected by cell counting kit-8 method to determine the effect concentration of antler peptide and dexamethasone.(2)MC3T3-E1 14 cells treated with dexamethasone(800 μmol/L)were intervened with different concentrations of gradient antler polypeptide,which were then divided into blank control group,dexamethasone group and dexamethasone+antler peptide group.Cell counting kit-8 method was used to calculate the effects of different concentrations of antler polypeptide on the proliferation of MC3T3-E1 14 cells.(3)Glutathione,superoxide dismutase,malondialdehyde,lipid peroxide,cellular iron,and reactive oxygen species levels in the blank control group,dexamethasone group and dexamethasone+antler peptide group were detected using kits.The protein expressions of glutathione peroxidase 4 and solute carrier family 7 member 11 were detected by western blot to verify the pathway by which antler polypeptide inhibits ferroptosis.RESULTS AND CONCIUSION:After cell activity was detected by cell counting kit-8 assay,antler peptide(10 mg/mL)and dexamethasone(800 μmol/L)were selected to treat MC3T3-E1 14 cells for 24 hours in subsequent experiments.After treatment with dexamethasone,malondialdehyde,lipid peroxide,cellular iron and reactive oxygen species levels were all increased(P<0.01),while glutathione content and superoxide dismutase activity were decreased and the protein expression of glutathione peroxidase 4 and solute carrier family 7 member 11 were also decreased(P<0.05-0.01).After antler peptide intervention,the changes in the above indexes were obviously reversed(P<0.05-0.01).To conclude,antler peptide may inhibit ferroptosis in osteoblasts by regulating the glutathione peroxidase 4/solute carrier family 7 member 11 axis,and thereby exert a therapeutic role in steroid-induced osteonecrosis of the femoral head.

Objective To investigate the role of Taohong Siwu Decoction(THSWD)in microglia pyroptosis mediated neuronal injury in PC12 under oxygen-glucose deprivation/reperfusion(OGD/R)and related mechanisms.Methods PC12 and BV2 cells were co-cultured using the OGD/R model to simulate ischemia-reperfusion injury of in vitro.The experiment was grouped as follows:control(PC12,PC12+BV2),OGD/R group(PC12,PC12+BV2),and OGD/R+10％THSWD-containing serum group(PC12,PC12+BV2).Oxygen and glucose was restored for 24h after 4-6h of deprivation.The severity of damage to PC12 cells was evaluated by cell counting kit-8(CCK-8)and flow cytometry.BV2 cells were used for the experiments and were divided into control,OGD/R and OGD/R+THSWD-containing serum(5％,10％,15％)groups,which were moulded and administered in the same way,and cell py-roptosis proteins proteinscysteinyl aspartate specific proteinase-1(caspase-1),apoptosis-associated speck-like protein containing a CARD(ASC),gasdermin D(GSDMD),interleukin-1β(IL-1β),interleukin-18(IL-18)and nucleotide-binding oligomeriza-tion domain-like receptor protein 3(NLRP3)inflammasome-associated proteins were detected by Western blot.Results In the pres-ence of BV2 cells,THSWD was able to further restore the morphology,increase cell viability(P＜0.05)and reduce apoptosis of PC12 cells after OGD/R injury(P＜0.05).In addition,THSWD was able to reduce the expression of OGD/R-induced pyroptosis proteins(caspase-1,ASC,GSDMD,IL-1β,IL-18)in BV2 microglia cells(P＜0.05).In contrast,the inhibitory effects of THSWD on BV2 cells pyroptosis proteins and NLRP3 inflammasome were reversed by NLRP3/caspase-1 agonist(P＜0.05).Conclusion THSWD protects PC 12 cells against OGD/R injury via inhibiting microglia pyroptosis mediated by the NLRP3/caspase-1 pathway.

Objective To evaluate the comprehensive value of methicillin-resistant Staphylococcus aureus(MRSA)treatment drugs in artificial joint infections from multiple perspectives and to solve the problem of MRSA infections in artificial joints.Methods Through literature research,relevant literature was retrieved and clinical studies meeting the requirements were selected and summarized.The Analytic Hierarchy Process(AHP)was applied to collect comprehensive clinical evaluation evidence and to conduct evaluations across different dimensions according to evaluation guidelines;the Delphi method combined with AHP was used for expert anonymous questionnaire evaluation,and the data was compared and analyzed.Results A clini-cal comprehensive evaluation index model for the treatment of artificial joint infections was successfully established,and the yaahp analysis software was used to score the comprehensive clinical evaluation evidence in various directions.Treatment drugs,vancomy-cin,daptomycin,and linezolid,at various levels were calculated through the software,and it was concluded that linezolid has the highest clinical comprehensive evaluation score.Conclusion Among the treatment drugs for MRSA infections in artificial joints,linezolid has a higher clinical comprehensive value and can provide a reference for the clinical treatment of MRSA infec-tions in artificial joints.