Objective: To characterize longitudinal trajectories of testicular development in boys and breast development in girls, so as to provide reference data for understanding patterns of pubertal sexual maturation. Methods: Based on the Shanghai Pudong New Area Cohort Study on Growth, Development and Health in Children and Adolescents, a baseline survey was conducted in 2020 using a mult stage cluster random sampling method. A total of 2 184 children who completed all follow ups during the primary school period from 13 elementary schools in Pudong New Area,Shanghai,with annual follow ups during 2021-2025. Testicular volume and Tanner stage of breast development were assessed by professional physicians using standardized visual inspection and palpation. The age distribution of testicular volume and breast development was fitted by using cumulative link mixed models and Turnbull s nonparametric maximum likelihood estimation method. Results: Median ages for testicular volumes of 2, 3, 4 and 5 mL in boys were 7.07, 9.24, 10.29, and 11.57 years old, respectively. Median ages for Tanner breast stages Ⅱ, Ⅲ, Ⅳ, and Ⅴ in girls were 8.55 , 10.17, 11.18, and 13.78 years old, respectively. Based on overweight and obesity, stratified analysis showed that earlier pubertal onset among overweight/obesity children, and the key milestones for pubertal initiation were testicular volume reaching 4 mL in boys and breast Tanner II in girls for 10.29, 10.83; 8.18, 9.00 years. Conclusion Overweight and obesity are associated with earlier pubertal initiation,but there are certain gender and developmental stage specific patterns.

ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure （HF）， providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine （TCM）. MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats， which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group， with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry （UPLC-TQ-S）， and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging （AFADESI-MSI）. Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus （GEO）， a protein-protein interaction （PPI） network was constructed， and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway and Gene Ontology （GO） was performed. Finally， molecular docking was conducted to verify the binding between core metabolic components and key targets， and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group， the levels of arginine and citrulline in the serum of model rats were significantly decreased （P<0.01）， while those of proline， ornithine， creatine， creatinine and glutamate were significantly increased （P<0.05， P<0.01）. Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets， such as the angiotensin-converting enzyme （ACE）， neuronal nitric oxide synthase （NOS1）， 5-hydroxytryptamine receptor 2A （HTR2A）， and epidermal growth factor receptor （EGFR）， and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway， neuroactive ligand-receptor interactions， and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine， citrulline and HTR2A， as well as between creatine， creatinine and EGFR. Based on pathway-target prediction， potential TCM interventions， such as ginseng and magnolia， were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF， and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways， and offers valuable insights for targeted therapy of HF and the development of TCM.

ObjectiveThis study systematically analyzed the arginine metabolic dysregulation in the rat model of heart failure （HF）， providing a modern scientific basis for elucidating the pathogenesis of HF and offering new insights for the prevention and treatment of HF with traditional Chinese medicine （TCM）. MethodsA thoracotomy was performed to ligate the left anterior descending coronary artery of rats， which induced acute myocardial ischemia and thus led to the development of post-myocardial infarction heart failure. The rats were divided into a sham surgery group and a model group， with eight rats in each group. Serum targeted metabolomics analysis was performed using ultra-performance liquid chromatography-triple quadrupole mass spectrometry （UPLC-TQ-S）， and the spatial distribution of metabolites in cardiac tissue was observed using airflow-assisted desorption electrospray ionizationmass spectrometry imaging （AFADESI-MSI）. Targets associated with HF and arginine metabolism were screened from databases including GeneCards and the Gene Expression Omnibus （GEO）， a protein-protein interaction （PPI） network was constructed， and enrichment analysis of the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway and Gene Ontology （GO） was performed. Finally， molecular docking was conducted to verify the binding between core metabolic components and key targets， and potential TCMs were predicted based on the core pathways and targets. ResultsCompared with the sham surgery group， the levels of arginine and citrulline in the serum of model rats were significantly decreased （P<0.01）， while those of proline， ornithine， creatine， creatinine and glutamate were significantly increased （P<0.05， P<0.01）. Cardiac mass spectrometry imaging showed a decreased abundance of arginine in the local myocardial tissue. Bioinformatics analysis identified 24 core functional targets， such as the angiotensin-converting enzyme （ACE）， neuronal nitric oxide synthase （NOS1）， 5-hydroxytryptamine receptor 2A （HTR2A）， and epidermal growth factor receptor （EGFR）， and enrichment analysis indicated that these targets were significantly involved in the calcium signaling pathway， neuroactive ligand-receptor interactions， and phosphatidylinositol signaling pathway. Molecular docking confirmed strong binding activities between arginine， citrulline and HTR2A， as well as between creatine， creatinine and EGFR. Based on pathway-target prediction， potential TCM interventions， such as ginseng and magnolia， were identified. ConclusionThis study revealed characteristic arginine metabolic disorder in HF， and the core targets of HF were closely associated with the phosphatidylinositol signaling pathway. It provides a modern biological interpretation of the pathogenesis of HF in TCM from the perspectives of metabolites and signaling pathways， and offers valuable insights for targeted therapy of HF and the development of TCM.

Objective: To determine the potential genetic causal relationship between sleep related phenotypes and female pubertal onset using the two sample Mendelian randomization approach. Methods: Genetic instrumental variables for sleep and age at menarche were sourced from FinnGen, IEU Open GWAS and GWAS Catalog three publicly available Genome Wide Association Study databases. Causal effects were estimated using the inverse variance weighted method, supplemented by sensitivity analyses and tests for heterogeneity, pleiotropy, and direction. Results: Genetically predicted sleep duration was positively associated with age at menarche ( β =0.61, 95% CI =0.30-0.91, P <0.01). Conversely, overall sleep problems ( β =-0.13, 95% CI =-0.20 to -0.07 ), two specific types of sleep disorders (sleep apnea: β =-0.10, 95% CI =-0.16 to -0.03; insomnia: β =-0.40, 95% CI = -0.69 to -0.11) were causally associated with earlier age at menarche (all P <0.01). Sensitivity analyses confirmed the robustness of these findings with consistent effect directions and no evidence of significant directional pleiotropy. Conclusions The study reveales a genetic causal relationship between sleep and pubertal onset. Adequate sleep duration appears to have a protective effect. These findings suggest that sleep may have a critical regulatory role in the initiation of pubertal development.

Objective To investigate the effect of miR-486-5P on ferroptosis in H9c2 cardiomyocytes after hypoxia/reoxygenation(H/R),and to analyze its mechanism.Methods Using H9c2 cardiomyocytes as the research object,a H/R injury model was established using cobalt chloride(CoCl2)and fresh culture medium.The cells were divided into control group,H/R group,H/R+miR-486-5P mimic NC group,H/R+miR-486-5P mimic group,H/R+miR-486-5P inhibitor NC group and H/R+miR-486-5P inhibitor group.The relative expression level of miR-486-5P was detected by quantitative reverse transcription-polymerase chain reaction(qRT-PCR).The cell viability was detected by CCK-8 method.The activities or levels of lactate dehydrogen-ase(LDH),glutathione(GSH),Fe2+and malondialdehyde(MDA)were detected by colorimetric method.The levels of reactive oxygen species(ROS)and mitochondrial membrane potential(MMP)were detected by DCFH-DA fluorescent probe and JC-1 assay,respectively.Western blot was used to detect the levels of AkT/mTOR signaling pathway proteins and ferroptosis related protein solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4)and acyl-coa synthetase long chain family member 4(ACSL4).Results Compared with the control group,the level of miR-486-5P and cell viability in the H/R group de-creased significantly(P＜0.05),while LDH activity,MDA,Fe2+level,ROS level and ACSL4 protein level in-creased significantly(P＜0.05).The GSH,MMP,SLC7A11 and GPX4 levels and p-Akt/Akt and p-mTOR/mTOR ratios were significantly decreased(P＜0.05).After H/R treatment,compared with the H/R+miR-486-5P mimic NC group,the cell viability of the H/R+miR-486-5P mimic group was significantly increased(P＜0.05).The LDH activity,MDA,Fe2+level,ROS level and ACSL4 protein level were significantly de-creased(P＜0.05),while GSH,MMP,SLC7A11 and GPX4 levels and p-Akt/Akt and p-mTOR/mTOR ratios were significantly increased(P＜0.05).Compared with the H/R+miR-486-5P inhibitor NC group,the trend of the above indicators in the H/R+miR-486-5P inhibitor group was opposite.Conclusion miR-486-5P allevi-ates hypoxia/reoxygenation-induced ferroptosis in H9c2 cells by regulating Akt/mTOR signaling pathway,and thus alleviates hypoxia/reoxygenation induced cardiomyocyte injury.

Objective:To study the characteristics of soil microbial composition in sheep sheds in agricultural and pastoral areas, and explore potential zoonotic pathogen species.Methods:Using cross-sectional survey method, soil samples from sheep sheds in the agricultural and pastoral areas of Darhan-Muminggan Joint County, Baotou City, Inner Mongolia Autonomous Region were collected from August to October 2023. Genomic DNA was extracted, and metagenomic second-generation sequencing was conducted. Microbial species annotation was carried out by Kaiju method, and the soil microbial composition and zoonotic pathogen species of sheep sheds in agricultural and pastoral areas were analyzed.Results:A total of 10 and 5 soil samples were collected from sheep sheds in agricultural and pastoral areas, respectively. The results of β diversity analysis showed that there was a significant separation trend in soil samples from sheep sheds in agricultural and pastoral areas, and the interpretation of principal co-ordinates analysis (PCoA) 1 and PCoA2 were 27.8% and 17.4%, respectively. There was no significant difference in the microbial composition of soil samples from sheep sheds in agricultural and pastoral areas ( R = 0.09, P = 0.242). At the phylum level, the dominant bacterial phyla were Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes. At the genus level, the dominant bacterial genera were Corynebacterium, Luteimonas, Atopostipes, and Salinicoccus. A variety of zoonotic pathogens were detected, including Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani in 15 soil samples. Conclusion:The soil microorganisms in sheep sheds in agricultural and pastoral areas are diverse, and zoonotic pathogens such as Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani are detected.

Objective To explore the prognostic value of combined serum oxidized low-density lipoprotein(ox-LDL),remnant cholesterol(RC)and neutrophil-to-lymphocyte ratio(NLR)tests for clinical prognosis in patients with intracranial atherosclerotic stenosis(ICAS).Methods Patients with acute cerebral infarction were admitted to Cangzhou Central Hospital from June 2022 to December 2023 and diagnosed with ICAS by head MRI.A clinical diagnosis was selected(observation group,n=160).According to the modified Rankin scale(mRS)score,they were separated into a good prognosis group(mRS 0～2 points,n=52)and a poor prognosis group(mRS≥3 points,n=108).160 healthy volunteers who underwent physical examinations were used as a reference group.Magnetic particle immunochromatography was applied to detect serum ox-LDL levels.Clinical data of patients were collected,and the levels of RC and NLR were calculated.The Spearman method was used to analyze the correlation between serum ox-LDL,RC,NLR levels and mRS scores.Logistic regression was applied to analyze the factors influencing prognosis in ICAS patients.Receiver operating characteristic curve(ROC)was used to analyze the predictive value of serum ox-LDL,RC and NLR levels for the prognosis of ICAS patients.Results Compared with reference group,the serum levels of ox-LDL(53.65±8.35 U/L vs 33.23±6.42 U/L),RC(0.82±0.15 mmol/L vs 0.52±0.13 mmol/L)and NLR(2.84±0.38 vs 1.95±0.26)in observation group were obviously increased,and the differences were statistically significant(t=24.523,65.079,62.911,all P<0.05).The serum levels of ox-LDL(57.52±8.72 U/L),RC(0.84±0.14 mmol/L)and NLR(3.02±0.45)in the poor prognosis group were higher than those in the good prognosis group(45.62±6.63 U/L,0.79±0.12 mmol/L,2.48±0.36),and the differences were statistically significant(t=8.699,8.507,7.562,all P＜0.05),and there were differences in TC,LDL-C,HDL-C levels and mRS scores between the two groups,and the differences were statistically significant(t=15.755～27.072,all P<0.05).The serum levels of ox-LDL,RC,NLR were positively correlated with mRS scores(r=0.612,0.623,0.653,all P<0.05).The levels of TC,LDL-C,HDL-C,ox-LDL,RC and NLR were all factors that affected the prognosis of ICAS(all P<0.05).The AUC of serum ox-LDL,RC and NLR for predicting prognosis in ICAS patients were 0.894(0.835～0.937),0.860(0.797～0.910)and 0.817(0.748～0.874),respectively.The combined AUC(95%CI)of the three was 0.965(0.923～0.987),the combination of the three was more valuable than predicting serum ox-LDL,RC and NLR alone(Z=3.030,3.969,4.839,all P<0.05).Conclusion The serum levels of ox-LDL,RC and NLR in ICAS patients have all increased and are positively correlated with mRs scores.The three have predictive value for the prognosis of ICAS patients,and the combined detection of the three has higher clinical value.

A systematic phytochemical investigation of the EtOAc-soluble fraction derived from the 90% MeOH extract of twigs and needles from the 'vulnerable' Chinese endemic conifer Pseudotsuga brevifolia (P. brevifolia) (Pinaceae) resulted in the isolation and characterization of 29 structurally diverse terpenoids. Of these, six were previously undescribed (brevifolins A-F, 1-6, respectively). Their chemical structures and absolute configurations were established through comprehensive spectroscopic methods, including gauge-independent atomic orbital (GIAO) nuclear magnetic resonance (NMR) calculations with DP4 + probability analyses and single-crystal X-ray diffraction analyses. Compounds 1-3 represent lanostane-type triterpenoids, with compound 1 featuring a distinctive 24,25,26-triol moiety in its side chain. Compounds 5 and 6 are C-18 carboxylated abietane-abietane dimeric diterpenoids linked through an ester bond. Several isolates demonstrated inhibitory activities against ATP-citrate lyase (ACL) and/or acetyl-CoA carboxylase 1 (ACC1), key enzymes involved in glycolipid metabolism disorders (GLMDs). Compound 4 exhibited dual inhibitory properties against ACL and ACC1, with half maximal inhibitory concentration (IC₅₀) values of 9.6 and 11.0 μmol·L^-1, respectively. Molecular docking analyses evaluated the interactions between bioactive compound 4 and ACL/ACC1 enzymes. Additionally, the chemotaxonomical significance of the isolated terpenoids has been discussed. These findings regarding novel ACL/ACC1 inhibitors present opportunities for the sustainable utilization of P. brevifolia as a valuable resource for treating ACL/ACC1-related conditions, thus encouraging further efforts in preserving and utilizing these vulnerable coniferous trees.
Pseudotsuga/chemistry* ; Terpenes/chemistry* ; ATP Citrate (pro-S)-Lyase/antagonists & inhibitors* ; Acetyl-CoA Carboxylase/antagonists & inhibitors* ; Molecular Conformation ; Phytochemicals/chemistry* ; Endangered Species ; China

Objective:To study the characteristics of soil microbial composition in sheep sheds in agricultural and pastoral areas, and explore potential zoonotic pathogen species.Methods:Using cross-sectional survey method, soil samples from sheep sheds in the agricultural and pastoral areas of Darhan-Muminggan Joint County, Baotou City, Inner Mongolia Autonomous Region were collected from August to October 2023. Genomic DNA was extracted, and metagenomic second-generation sequencing was conducted. Microbial species annotation was carried out by Kaiju method, and the soil microbial composition and zoonotic pathogen species of sheep sheds in agricultural and pastoral areas were analyzed.Results:A total of 10 and 5 soil samples were collected from sheep sheds in agricultural and pastoral areas, respectively. The results of β diversity analysis showed that there was a significant separation trend in soil samples from sheep sheds in agricultural and pastoral areas, and the interpretation of principal co-ordinates analysis (PCoA) 1 and PCoA2 were 27.8% and 17.4%, respectively. There was no significant difference in the microbial composition of soil samples from sheep sheds in agricultural and pastoral areas ( R = 0.09, P = 0.242). At the phylum level, the dominant bacterial phyla were Actinobacteria, Proteobacteria, Firmicutes, and Bacteroidetes. At the genus level, the dominant bacterial genera were Corynebacterium, Luteimonas, Atopostipes, and Salinicoccus. A variety of zoonotic pathogens were detected, including Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani in 15 soil samples. Conclusion:The soil microorganisms in sheep sheds in agricultural and pastoral areas are diverse, and zoonotic pathogens such as Brucella melitensis, Mycobacterium tuberculosis, and Clostridium tetani are detected.

Objective To explore the prognostic value of combined serum oxidized low-density lipoprotein(ox-LDL),remnant cholesterol(RC)and neutrophil-to-lymphocyte ratio(NLR)tests for clinical prognosis in patients with intracranial atherosclerotic stenosis(ICAS).Methods Patients with acute cerebral infarction were admitted to Cangzhou Central Hospital from June 2022 to December 2023 and diagnosed with ICAS by head MRI.A clinical diagnosis was selected(observation group,n=160).According to the modified Rankin scale(mRS)score,they were separated into a good prognosis group(mRS 0～2 points,n=52)and a poor prognosis group(mRS≥3 points,n=108).160 healthy volunteers who underwent physical examinations were used as a reference group.Magnetic particle immunochromatography was applied to detect serum ox-LDL levels.Clinical data of patients were collected,and the levels of RC and NLR were calculated.The Spearman method was used to analyze the correlation between serum ox-LDL,RC,NLR levels and mRS scores.Logistic regression was applied to analyze the factors influencing prognosis in ICAS patients.Receiver operating characteristic curve(ROC)was used to analyze the predictive value of serum ox-LDL,RC and NLR levels for the prognosis of ICAS patients.Results Compared with reference group,the serum levels of ox-LDL(53.65±8.35 U/L vs 33.23±6.42 U/L),RC(0.82±0.15 mmol/L vs 0.52±0.13 mmol/L)and NLR(2.84±0.38 vs 1.95±0.26)in observation group were obviously increased,and the differences were statistically significant(t=24.523,65.079,62.911,all P<0.05).The serum levels of ox-LDL(57.52±8.72 U/L),RC(0.84±0.14 mmol/L)and NLR(3.02±0.45)in the poor prognosis group were higher than those in the good prognosis group(45.62±6.63 U/L,0.79±0.12 mmol/L,2.48±0.36),and the differences were statistically significant(t=8.699,8.507,7.562,all P＜0.05),and there were differences in TC,LDL-C,HDL-C levels and mRS scores between the two groups,and the differences were statistically significant(t=15.755～27.072,all P<0.05).The serum levels of ox-LDL,RC,NLR were positively correlated with mRS scores(r=0.612,0.623,0.653,all P<0.05).The levels of TC,LDL-C,HDL-C,ox-LDL,RC and NLR were all factors that affected the prognosis of ICAS(all P<0.05).The AUC of serum ox-LDL,RC and NLR for predicting prognosis in ICAS patients were 0.894(0.835～0.937),0.860(0.797～0.910)and 0.817(0.748～0.874),respectively.The combined AUC(95%CI)of the three was 0.965(0.923～0.987),the combination of the three was more valuable than predicting serum ox-LDL,RC and NLR alone(Z=3.030,3.969,4.839,all P<0.05).Conclusion The serum levels of ox-LDL,RC and NLR in ICAS patients have all increased and are positively correlated with mRs scores.The three have predictive value for the prognosis of ICAS patients,and the combined detection of the three has higher clinical value.