arcopenia is a systemic skeletal muscle disorder characterized by a decrease in skeletal muscle mass and progressive decline in function， with multiple signaling pathways involved in its occurrence and development. Among them， the AMP-activated protein kinase （AMPK） signaling pathway， as a key pathway regulating cellular energy homeostasis， plays an important role in the regulation of skeletal muscle metabolism and functional maintenance by improving abnormalities in glucose and lipid metabolism， balancing skeletal muscle protein synthesis and degradation， improving mitochondrial function， promoting autophagy， and inhibiting inflammatory responses and oxidative stress. This article reviews the research progress on how various traditional Chinese medicine （TCM） monomers， including polyphenols， flavonoids， and terpenoids； various traditional Chinese medicine extracts， such as those from Lycium barbarum ， Asini Corii Colla， and Panax quinquefolium ， and TCM compounds， such as Guiqi zhuangjin decoction， Jianpi qiangji granules， and Qigu capsules， intervene in sarcopenia by regulating the AMPK signaling pathway to promote muscle protein synthesis， inhibit protein degradation， improve mitochondrial function， and alleviate inflammation and oxidative stress. Additionally， their molecular mechanisms are explored. The aim is to deeply elucidate the basis of TCM in the prevention and treatment of sarcopenia and to provide theoretical support for the development of related innovative drugs.

Pigeon circovirus(PiCV)is globally widespread and is considered a potential cause of young pigeon sickness syndrome(YPDS),which leads to severe immunosuppression and high mortality.Due to the inability of PiCV to be cultured in cells,the development of traditional vac-cines is severely limited,and no effective vaccines is currently available.To develop a novel PiCV DNA candidate vaccine,we cloned the △Cap gene lacking a nuclear localization signal(NLS),and fused it at its C-terminus with the transmembrane and cytoplasmic regions of the Newcastle dis-ease virus(NDV)F protein(△Cap-TMCT).Two DNA vaccine candidates were constructed:pCAGG-△Cap,targeting intracellular expression,and pCAGG-△Capt,for cell surface expression,respectively.The results of indirect immunofluorescence and Western blot analyses confirmed suc-cessful expression of both recombinant plasmids in DF1 cells.Immunization studies in mice re-vealed that pCAGG-△Capt induced significantly higher levels of specific IgG antibodies,T-cell re-sponses,and cytokine secretion compared to pCAGG-△Cap,as assessed by ELISA,flow cytome-try,and ELISpot assays.These findings suggest that targeting △Cap-TMCT fusion protein to the cell surface can effectively enhance its immunogenicity,highlighting its potential as a PiCV DNA vaccine candidate.This study provides new strategies and theoretical foundations for the design and development of PiCV DNA vaccines.

Objective:To investigate the inhibitory effect and mechanism of puerarin(Pue)on hydrogen peroxide(H2 O2)induced ferroptosis in the rat Schwann cell derived cell line RSC96.Methods:The RSC96 cells were incuba-ted with H2 O2 to establish a cellular injury model,while a subset of cells were co-incubated with H2 O2 and Pue.The vi-ability of cells was examined using the CCK-8 assay.The intracellular levels of glutathione(GSH),total superoxide dismutase(T-SOD),reactive oxygen species(ROS),malondialdehyde(MDA),and ferrous ions(Fe2+)levels were quantified using commercial kits.Western blot was employed to detect the protein expression level of glutathione peroxi-dase 4(GPX4),cyclooxygenase-2(COX-2),solute carrier family 7 member 11(SLC7A11),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1).Immunofluorescence assay was performed to examine the expression and nuclear distribution of Nrf2.Results:Pue pretreatment significantly increased the survival rate of H2 O2-treated RSC96 cells,increased the intracellular content of GSH and T-SOD,and inhibited the concentration of ROS and MDA.It also activated the nuclear translocation of Nrf2 and upregulated GPX4,SLC7A11,HO-1,and Nrf2 proteins in RSC96 cells;This effect was abolished by the Nrf2 inhibitor ML385.Conclusion:Pue treatment alleviated the H2 O2-induced ferroptosis of RSC96 cells via the Nrf2/SLC7A11/GPX4 signaling pathway.

Objective:To investigate the targets and mechanisms of cycloastragenol in ameliorating azithromycin-induced drug-induced liver injury(DILI)based on network pharmacology and in vitro experiment validation.Methods:Potential targets of cycloastragenol and DILI were predicted using databases.The common and key targets were screened and subjected to GO and KEGG enrichment analyses,as well as molecular docking validation.Primary hepatocytes from C57BL/6 mice were isolated.The optimal concentration and time for azithromycin-induced DILI in mouse primary hepatocytes were determined using CCK8 and ROS assays.The expression of genes and proteins such as NF-κB p65,p-NF-κB p65,AMPKα,and p-AMPKα was assessed using RT-qPCR and Western blot to evaluate the intervention effect of cycloastragenol(10-50 μmol/L).Results:Network pharmacology analysis identified 10 key genes related to cycloastragenol's improvement of DILI,including heat shock protein 90AA1(HSP90AA1),matrix metalloproteinase 2(MMP2),etc.GO enrichment analysis suggested that cycloastragenol primarily regulates biological processes such as membrane potential and chemical synaptic transmission,and affects cellular components such as neuronal cell bodies and distal axons,and related kinase activities.KEGG enrichment analysis showed that it mainly exerts intervention effects through neuro-signaling pathways and IL-17 signaling pathways.Molecular docking demonstrated strong binding of cycloastragenol to HSP90AA1,MMP2,NF-κB p65,AMPKα,nuclear factor erythroid 2-related factor 2(Nrf2),heme oxygenase 1(HO-1),and NAD(P)H:quinone oxidoreductase 1(NQO1),with a binding energy≤-5.0 kcal/mol for Nrf2.In vitro experiments showed that azithromycin(50 μmol/L,12 h)significantly reduced hepatocyte viability and increased ROS levels(P＜0.01).Different concentrations of cycloastragenol significantly improved the activity of mouse primary hepatocytes,reduced the generation of intracellular ROS,downregulated the phosphorylation level of NF-κB p65,and upregulated the mRNA and protein levels of AMPKα,Nrf2,HO-1,NQO1(P＜0.05).Conclusions:Cycloastragenol may alleviate azithromycin-induced hepatocyte oxidative stress and inflammation by inhibiting NF-κB phosphorylation and activating the AMPK/Nrf2/HO-1/NQO1 pathway,with its mechanism likely closely linked to targeting Nrf2.However,the complex mechanisms of DILI may involve additional unverified pathways.Therefore,further studies are necessary to validate the efficacy and safety of cycloastragenol in animal models.

Objective Network pharmacology and molecular docking techniques were used to predict the potential mechanisms by which the active components of Piper nigrum(PN)regulate depressive-like behaviors in chronic restraint stress(CRS)mice.Methods The major chemical components and targets of PN were screened using the Traditional Chinese Medicine Systems Pharmacology database.Targets related to ferroptosis and depression were obtained from the Online Mendelian Inheritance in Man,GeneCards,and FerrDB databases.The intersecting targets were then subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Gnomes(KEGG)pathway enrichment analyses,and molecular docking was performed to validate the binding capacities between the core targets and their corresponding active components.Finally,we established a CRS mouse model.Mice were treated with PN 75,150,and 300 mg/kg for 4 weeks,followed by behavioral assessments and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)to verify the expression of core genes.Results Nine active components were screened from PN,corresponding to 27 targets,and 8377 targets related to depression and 547 targets associated with ferroptosis were screened from the databases.The intersection of these three sets resulted in 25 target genes.KEGG enrichment analysis revealed that these core targets were predominantly enriched in signaling pathways,including cholinergic synapses,serotonergic synapses,and neuroactive ligand-receptor interactions.Molecular docking result showed that the main active components of PN had strong binding affinities for the targets CHRM2,SLC6A4,PTGS2,and SLC6A2.Behavioral assessments demonstrated that PN significantly increased the sucrose preference index(P＜0.01,P＜0.001),reduced immobility time in the tail suspension and forced swimming tests(P＜0.01,P＜0.001),and enhanced exploratory behavior in the open field test(P＜0.05.P＜0.01,P＜0.001).PN significantly reduced the serum levels of inflammation markers(P＜0.05.P＜0.01,P＜0.001),as shown by enzyme-linked immunosorbent assay,and neurotransmitter analysis revealed that PN significantly increased the levels of serotonin and acetylcholine in the mouse hippocampus(P＜0.05).RT-qPCR showed that PN demonstrated the mRNA expression of SLC6A4(P＜0.05.P＜0.01,P＜0.001).Conclusions PN may improve depressive-like behavior in mice by modulating serotonin and acetylcholine levels,inhibiting inflammatory responses,participating in immune regulation,and exerting neuroprotective effects.

Objective:To compare the cumulative live birth rate (CLBR) per oocyte retrieval cycle between gonadotropin-releasing hormone agonist (GnRH-a) long protocol and GnRH antagonist (GnRH-A) protocol in overweight and obese women undergoing in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). Methods:A retrospective cohort study was conducted to analyze the clinical characteristics of overweight and obese patients who underwent IVF/ICSI at the Center of Reproductive Medicine, the First Affiliated Hospital of Nanjing Medical University between January 2013 and December 2019. A total of 3 707 cycles were executed in overweight and obese patients who fulfilled the prescribed inclusion criteria, comprising 1 555 GnRH-a long protocol cycles and 2 152 GnRH-A protocol cycles. To mitigate confounding factors, post hoc randomization and propensity score matching (PSM) at a 1∶1 ratio were applied to match female age, anti-Müllerian hormone levels, and antral follicle count. The primary outcome observation indicator was the CLBR of the oocyte retrieval cycle. Analysis of subgroups of the population was conducted by the women's body mass index, age, and polycystic ovarian syndrome (PCOS) status.Results:After PSM, a total of 2 496 cycles were included comprising 1 248 GnRH-a long protocol cycles and 1 248 GnRH-A protocol cycles. GnRH-a long protocol had a higher CLBR [71.88% (897/1 248)] than that in GnRH-A protocol [62.98% (786/1 248), P<0.001]. No statistically significant difference was observed in the interval from gonadotropin initiation to live birth delivery day between the GnRH-a long protocol and GnRH-A protocol ( P>0.05). Subgroup analysis revealed that after PSM, the CLBR of GnRH-a long protocol in the patients with a body mass index of 25.0-29.9 kg/m 2 [71.36% (856/1 195)] and ≥30.0 kg/m 2 [77.36% (41/53)] were higher than those of the GnRH-A protocol patients [63.30% (759/1 199), P<0.001; 55.10% (27/49), P=0.017]. The CLBR of GnRH-a long protocol in women aged 20-34 [73.32% (805/1 098)] and ≥35 years [61.33% (92/150)] were higher than those of the GnRH-A protocol patients [67.18% (696/1 036), P=0.002; 42.45% (90/212), P<0.001]; among patients without PCOS, the CLBR with the GnRH-a long protocol [71.55% (850/1 188)] was significantly higher than that with GnRH-A protocol [60.95% (654/1 073), P<0.001]. However, in overweight and obese patients with PCOS, there was no statistically significant difference in CLBR between the two protocols ( P>0.05). The incidence of moderate-severe ovarian hyperstimulation syndrome (OHSS) was significantly lower in the overweight and obese population using GnRH-A protocol [0.64% (8/1 248)] compared with GnRH-a long protocol [1.76% (22/1 248), P=0.016]. Conclusion:For overweight and obese patients, GnRH-a long protocol demonstrates higher CLBR compared with GnRH-A protocol, indicating superior efficacy. For those with PCOS, both protocols show comparable CLBR, while the incidence of severe OHSS is lower in the GnRH-A.

Objective:To evaluate the cumulative live birth rate (CLBR) and cost-effectiveness of fixed versus adjusted follicle-stimulation hormone (FSH) dosages in infertile women with different ovarian responses during their first assisted reproductive technology (ART) cycle.Methods:A retrospective real-world cohort study was conducted on 5 419 infertile women who underwent their first ART treatment at the Department of Reproductive Medicine of the First Affiliated Hospital of Nanjing Medical University between January 2013 and December 2017. All patients received an individualized starting dosage of gonadotropin. Based on whether FSH dosages were adjusted during controlled ovarian stimulation (COS), patients were divided into fixed-dosage group ( n=2 061) and adjusted-dosage group ( n=3 358). Clinical outcomes and FSH cost-effectiveness were compared between the two groups across different ovarian response groups, with CLBR as the primary outcome. Propensity score matching (PSM) and multivariable logistic regression were used to adjust for potential confounders. Results:FSH dosage adjustments were found in 62.0% (3 358/5 419) of cycles during COS. After PSM, baseline characteristics were comparable between the two groups (all P>0.05). After adjusting for confounders using multivariable logistic regression, FSH dosage adjustment was not significantly associated with CLBR ( OR=1.06, 95% CI: 0.94-1.20, P=0.332). Compared with the adjusted-dosage group, the fixed-dosage group showed no significant differences in CLBR in poor-, normal-, and high-responder groups (all P>0.05). The incidence of ovarian hyperstimulation syndrome (OHSS) did not differ significantly between the two groups ( P>0.05). In poor-, normal-, and high-responder groups, the total FSH dosages in the fixed-dose group [1 350 (375, 1 825) U, 1 200 (375, 1 500) U and 525 (375, 1 128) U, respectively] were significantly lower than those in the adjusted-dose group [1 875 (1 425, 2 294) U, P=0.001; 1 425 (450, 1 875) U, P<0.001; 600 (375, 1 425) U, P=0.020]. Similarly, average FSH costs in different ovarian response groups in the fixed-dosage group [4 725.0 (1 312.5, 6 387.5) yuan, 4 200.0 (1 312.5, 5 250.0) yuan and 1 837.5 (1 312.5, 3 947.3) yuan, respectively] were significantly lower than those in the adjusted-dosage group [6 562.5 (4 987.5, 8 028.1) yuan, P=0.001; 4 987.5 (1 575.0, 6 562.5) yuan, P<0.001; 2 100.0 (1 312.5, 4 987.5) yuan, P=0.020]. For normal-responders, the FSH cost per high-quality embryo in the fixed-dosage group [1 365.0 (875.0, 2 537.5) yuan] was significantly lower than that in the adjusted-dosage group [2 056.3 (1 268.8, 3 412.5) yuan, P<0.001]. Conclusion:FSH dosage adjustment during COS is not associated with CLBR or the incidence of OHSS. However, the fixed-dose group exhibited lower total FSH dosages and costs across different ovarian response populations. In the context of ART being covered by medical insurance, fixed FSH dosage may represent a more cost-effective ovarian stimulation protocol.

Objective:To investigate the inhibitory effect and mechanism of puerarin(Pue)on hydrogen peroxide(H2 O2)induced ferroptosis in the rat Schwann cell derived cell line RSC96.Methods:The RSC96 cells were incuba-ted with H2 O2 to establish a cellular injury model,while a subset of cells were co-incubated with H2 O2 and Pue.The vi-ability of cells was examined using the CCK-8 assay.The intracellular levels of glutathione(GSH),total superoxide dismutase(T-SOD),reactive oxygen species(ROS),malondialdehyde(MDA),and ferrous ions(Fe2+)levels were quantified using commercial kits.Western blot was employed to detect the protein expression level of glutathione peroxi-dase 4(GPX4),cyclooxygenase-2(COX-2),solute carrier family 7 member 11(SLC7A11),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1).Immunofluorescence assay was performed to examine the expression and nuclear distribution of Nrf2.Results:Pue pretreatment significantly increased the survival rate of H2 O2-treated RSC96 cells,increased the intracellular content of GSH and T-SOD,and inhibited the concentration of ROS and MDA.It also activated the nuclear translocation of Nrf2 and upregulated GPX4,SLC7A11,HO-1,and Nrf2 proteins in RSC96 cells;This effect was abolished by the Nrf2 inhibitor ML385.Conclusion:Pue treatment alleviated the H2 O2-induced ferroptosis of RSC96 cells via the Nrf2/SLC7A11/GPX4 signaling pathway.

Objective Network pharmacology and molecular docking techniques were used to predict the potential mechanisms by which the active components of Piper nigrum(PN)regulate depressive-like behaviors in chronic restraint stress(CRS)mice.Methods The major chemical components and targets of PN were screened using the Traditional Chinese Medicine Systems Pharmacology database.Targets related to ferroptosis and depression were obtained from the Online Mendelian Inheritance in Man,GeneCards,and FerrDB databases.The intersecting targets were then subjected to Gene Ontology and Kyoto Encyclopedia of Genes and Gnomes(KEGG)pathway enrichment analyses,and molecular docking was performed to validate the binding capacities between the core targets and their corresponding active components.Finally,we established a CRS mouse model.Mice were treated with PN 75,150,and 300 mg/kg for 4 weeks,followed by behavioral assessments and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)to verify the expression of core genes.Results Nine active components were screened from PN,corresponding to 27 targets,and 8377 targets related to depression and 547 targets associated with ferroptosis were screened from the databases.The intersection of these three sets resulted in 25 target genes.KEGG enrichment analysis revealed that these core targets were predominantly enriched in signaling pathways,including cholinergic synapses,serotonergic synapses,and neuroactive ligand-receptor interactions.Molecular docking result showed that the main active components of PN had strong binding affinities for the targets CHRM2,SLC6A4,PTGS2,and SLC6A2.Behavioral assessments demonstrated that PN significantly increased the sucrose preference index(P＜0.01,P＜0.001),reduced immobility time in the tail suspension and forced swimming tests(P＜0.01,P＜0.001),and enhanced exploratory behavior in the open field test(P＜0.05.P＜0.01,P＜0.001).PN significantly reduced the serum levels of inflammation markers(P＜0.05.P＜0.01,P＜0.001),as shown by enzyme-linked immunosorbent assay,and neurotransmitter analysis revealed that PN significantly increased the levels of serotonin and acetylcholine in the mouse hippocampus(P＜0.05).RT-qPCR showed that PN demonstrated the mRNA expression of SLC6A4(P＜0.05.P＜0.01,P＜0.001).Conclusions PN may improve depressive-like behavior in mice by modulating serotonin and acetylcholine levels,inhibiting inflammatory responses,participating in immune regulation,and exerting neuroprotective effects.

Objective To determine the serum level of quantitative hepatitis B core antibody(qAnti-HBc)in decompensated hepatitis B cirrhosis patients with normal international normalized ratio(INR),and to investigate its prognostic value in this target population.Methods A total of 120 decompensated hepatitis B cirrhosis patients with normal INR who were diagnosed and treated in Tianjin Third Central Hospital from October 1,2018 to April 1,2021 were enrolled.Baseline indicators were collected,the serum level of qAnti-HBc was measured,and the prognosis was followed up.According to the prognosis,the patients were divided into survival group and death group.The independent-samples t test or the Mann-Whitney U test was used for comparison of continuous data between two groups,and the chi-square test was used for comparison of categorical data between groups.Univariate and multivariate Cox regression analyses were used to investigate the candidate variables affecting the prognosis and establish a prognostic model for this population,and the receiver operating characteristic(ROC)curve was plotted.Results The patients enrolled were followed up for 32.17±13.09 months,and there were 99 patients(82.5%)in the survival group and 21 patients(17.5%)in the death group.There were significant differences between the survival group and the death group in sex(χ2=2.151,P=0.014),age(t=-3.218,P=0.003),total bilirubin(Z=-0.901,P=0.027),albumin(t=3.353,P=0.001),and Child-Pugh class(χ2=1.144,P=0.010).The univariate and multivariate Cox regression analyses showed that serum qAnti-HBc(hazard ratio[HR]=0.57,95%confidence interval[CI]:0.32-1.00,P=0.043),age(HR=1.06,95%CI:1.00-1.12,P=0.044),sex(HR=3.82,95%CI:1.46-10.00,P=0.006),platelet count(HR=0.98,95%CI:0.97-1.00,P=0.037),and albumin(HR=0.86,95%CI:0.79-0.95,P=0.002)were independent risk predictive factors for the prognosis of decompensated hepatitis B cirrhosis patients with normal INR.Based on these factors,a predictive model was established as h(t,x)/h0(t)=exp(1.34X1+0.06X2-0.14X3-0.02X4-0.57X5),where X1=sex,X2=age,X3=albumin,X4=platelet count,X5=qAnti-HBc.This predictive model had an area under the ROC curve of 0.842,with a sensitivity of 0.79,a specificity of 0.73,and a C-index of 0.85.Conclusion In decompensated hepatitis B cirrhosis patients with normal INR(0.8-1.2),serum qAnti-HBc is one of the independent risk predictive factors for death,and the predictive model established based on the combination of serum qAnti-HBc and other indicators can effectively predict the prognosis of patients.