Eight compounds were isolated and purified from the ethyl acetate part of 70% acetone extract of Rehmannia glutinosa by various chromatographic techniques such as silica gel, MCI gel CHP-20, ODS, Toyopearl HW-40C, combined with TLC and semi-preparative HPLC. Their structures were elucidated by modern spectroscopy techniques (NMR, MS, UV, IR), and identified as neomartynoside A (1), osmanthuside B₆ (2), martynoside (3), isomartynoside (4), (E)-p-hydroxycinnamic acid (5), caffeic acid (6), ferulic acid (7), and methyl caffeate (8). Compound 1 is a new phenylethanol glycoside, which was identified as neomartynoside A. Compound 2 was isolated from Rehmannia glutinosa for the first time. In addition, compounds 2, 6 and 7 significantly increased relative glucose consumption, showing potential hypoglycemic activity.

Aim To investigate the intervention effect of Rehmannia radix water extract on bleomycin(BLM)-induced pulmonary fibrosis in mice combined with metabolomics and to reveal the potential mechanism,in order to provide new ideas for clinical treatment of pul-monary fibrosis.Methods Male C57BL/6N mice were randomly divided into the control group,model group,pirfenidone group(positive control,PFD,270 mg·kg-1),and low dose(DH-L,4.55 g·kg-1)group,medium dose(DH-M,9.1 g·kg-1)group and high dose(DH-H,18.2 g·kg-1)group of Rehman-nia.Except for the control group,BLM(5 mg·kg-1)was instilled into the trachea to establish the model of pulmonary fibrosis in the other groups.The survival rate,lung index and blood oxygen saturation of mice in each group were evaluated.HE and Masson staining were used to observe the pathological changes of lung tissue.WBP was used to detect lung function.Flow cytometry was used to detect the apoptosis of primary lung cells,ROS and immune cells.ELISA was used to detect the levels of fibrosis markers and inflammatory factors(α-SMA,collagen Ⅰ,collagen Ⅲ,TGF-β1,TNF-α,IL-1 β,and IL-6).Biochemical method was employed to detect the contents of GSH-Px,T-SOD and MDA.Liquid chromatograph mass spectrometer(LC-MS)metabolomics was used to analyze the changes of serum metabolic profile.Results Water extract of Re-hmannia significantly increased the survival rate,oxy-gen saturation and lung function of mice with pulmona-ry fibrosis,reduced the lung coefficient,ameliorated pathological damage and collagen deposition in lung tissue,reduced the levels of apoptosis and oxidative stress,and down-regulated the levels of inflammatory factors in lung tissue.It regulated the levels of metabo-lites such as bile acid metabolism,sphingolipid metabo-lism,and unsaturated fatty acid metabolism.Conclu-sions Water extract of Rehmannia inhibits lung injury and collagen deposition in mice with pulmonary fibrosis by inhibiting inflammatory response,which may be a-chieved by regulating the levels of inflammatory factors through the metabolic pathways of bile acid and sphin-golipid.

Interleukin- (IL) 23 drives pathogenic differentiation of Th17 cells via the JAK2-STAT3 signaling pathway, upregulates IL-17A/IL-22 expression, and disrupts intestinal barrier integrity, thereby playing a pivotal role in the pathogenesis of Crohn's disease (CD). IL-23p19 inhibitors—exemplified by risankizumab, mirikizumab, and guselkumab—precisely block this pathway. Key phase 3 trials have demonstrated their efficacy in CD, showing significant clinical benefits in patients refractory to conventional therapies or biologics, with no new safety signals identified. The ultimate treatment goal for CD is deep healing (mucosal-transmural-biochemical composite remission) as defined by STRIDE II. However, current evidence exhibits critical limitations: absence of head-to-head drug comparisons, insufficient data on biologic-experienced subpopulations, and heterogeneous follow-up durations leading to uncertainties in long-term safety. Future studies require standardized head-to-head trials with enhanced subgroup analyses to optimize precision therapeutics.

To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

Objective:To investigate the protective effect of achyranthes bidentata(AB)on sperm quality in mice with sper-matogenic disorder through the glycolytic metabolic pathway and its action mechanism.Methods:We equally randomized 40 Kun-ming mice into a normal control,a model control,a low-dose AB(3.5 g/kg)and a high-dose AB group(7.0 g/kg),and established the model of spermatogenic disorder in the latter three groups of mice by intraperitoneal injection of doxorubicin(30 mg/kg).Two days after modeling,we collected the testis and kidney tissues and blood samples from the mice for observation of the pathological changes in the testis tissue by HE staining,detection of perm motility with the sperm quality analyzer,examination of the apoptosis of testis cells by flow cytometry,measurement of the levels of testosterone(T),malondialdehyde(MDA),superoxide dismutase(SOD)and cata-lase(CAT)in the serum and testis tissue by ELISA,and determination of expressions of the key enzymes of glycolysis hexokinase Ⅱ(HK2),pyruvate kinase M2(PKM2),platelet phosphofructokinase(PFKP),lactate dehydrogenase A(LDHA)and the meiosis pro-teins REC8 and SCP3 by Western blot,and the mRNA expressions of glycolytic phosphofructokinase 1(PFK1),phosphoglycerate ki-nase 1(PGK1),tumor necrosis factor-α(TNF-α)and interleukin-1 β(IL-1β)by fluorescence quantitative PCR(FQ-PCR).Results:Compared with the model controls,the mice in the AB groups showed significant increases in the testis coefficient,kidney in-dex,sperm concentration,sperm motility,spermatogonia,primary spermatocytes,spermatids,sperm count and the serum T level(P＜0.05 orP＜0.01),but dramatic decreases in the apoptosis of testis cells and percentage of morphologically abnormal sperm(P＜0.01).Achyranthes bidentata also significantly elevated the levels of SOD and CAT,and down-regulated the mRNA expressions of MDA,TNF-α and IL-1β(P＜0.05 or P＜0.01),and up-regulated the protein expressions of HK2,PKM2,PFKP,LDHA,REC8 and SCP3,and expressions of the glycolysis key genes Pfk1 and Pgk1(P＜0.05 orP＜0.01).Conclusion:Achyranthes bidentata ameliorates doxorubicin-induced spermatogenic disorder in mice by regulating the glycolytic pathway and reducing oxidative stress and the expressions of inflammatory factors.

Interleukin- (IL) 23 drives pathogenic differentiation of Th17 cells via the JAK2-STAT3 signaling pathway, upregulates IL-17A/IL-22 expression, and disrupts intestinal barrier integrity, thereby playing a pivotal role in the pathogenesis of Crohn's disease (CD). IL-23p19 inhibitors—exemplified by risankizumab, mirikizumab, and guselkumab—precisely block this pathway. Key phase 3 trials have demonstrated their efficacy in CD, showing significant clinical benefits in patients refractory to conventional therapies or biologics, with no new safety signals identified. The ultimate treatment goal for CD is deep healing (mucosal-transmural-biochemical composite remission) as defined by STRIDE II. However, current evidence exhibits critical limitations: absence of head-to-head drug comparisons, insufficient data on biologic-experienced subpopulations, and heterogeneous follow-up durations leading to uncertainties in long-term safety. Future studies require standardized head-to-head trials with enhanced subgroup analyses to optimize precision therapeutics.

To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

Chemiluminescence,as a self-luminous phenomenon that does not require light,heat,acoustic,electric and magnetic excitation,has been widely used in the fields of analytical chemistry,cold light source and bio-imaging because of its advantages including high sensitivity,wide linear range,simple equipment and fast detection speed compared with other analytical techniques.Carbon dots(CDs)are a class of nanomaterials with excellent photoluminescence properties and high biocompatibility.CDs are stable,easy to prepare and abundant in types,and researchers have introduced many types of CDs into different chemiluminescence systems.In this paper,the applications of CDs in common chemiluminescence systems and the possible mechanisms of action were discussed,and the research progresses on the application of CDs in different chemiluminescence detection fields in recent years were summarized.Finally,the development trend of CDs in chemiluminescence was analyzed.

Poly(lactic-co-glycolide)(PLGA)is a key excipient in long-acting sustained-release preparations,and its degradation properties directly affect the drug release behavior.In this study,PLGA microspheres were prepared by microfluidic techniques,and the morphology changes of the microspheres were observed by scanning electron microscopy(SEM).In alkaline environment,due to the accelerated hydrolysis of ester bonds,the surface of the microspheres was rapidly dissolved and eroded,and the degradation rate was significantly higher than that in acidic environment.High temperature accelerated the degradation of PLGA microspheres.Under neutral and alkaline conditions,the microspheres showed aggregation and adhesion.Under acidic conditions,the microspheres gradually decomposed into irregular fragments.The high ionic strength further promoted the surface corrosion of the microspheres,especially under extreme pH conditions.Simultaneously,PLGA microspheres encapsulating coumarin were prepared to simulate the microsphere formulation.The release rate of coumarin after degradation of the microspheres under different conditions was observed by measuring the absorbance with ultraviolet-visible spectrophotometry.The results were consistent with those of the blank microspheres.This study revealed that the degradation of PLGA microspheres was significantly pH-dependent,temperature sensitive and ion strength responsive.These findings not only helped to understand and optimize the long-term stability and controlled release performance of drug-carrying microspheres,but also provided a theoretical basis for further improvement of PLGA-based drug carrier design.

Objective:To explore the effects and mechanisms of Shen-su-yin (SSY) on acute lung injury (ALI) in rats based on untargeted Metabolomics, network pharmacology, and experimental verification.Methods:Untargeted Metabolomics was performed to detect the ingredients of SSY by using ultra-high performance liquid chromatography-Q-exactive orbitrap mass spectrum, and the active ingredients were screened from the detected ingredients. Common targets of the active ingredient targets and ALI targets were utilized to screen hub targets to perform gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. Then, key hub targets were selected from the hub targets, and the active ingredients-hub targets network was built to screen core ingredients. Subsequently, molecular docking was performed between the key hub targets and the core ingredients. 48 rats were randomly and equally divided into 4 groups by using a random number table: normal control group, lipopolysaccharide-induced ALI group, ALI+SSY group, and ALI+dexamethasone group. 24 hours after lipopolysaccharide induction, the levels of respiratory rate, blood lactate, lung wet/dry weight ratio, ALI score, inflammatory factors of bronchoalveolar lavage fluid, and oxidative stress mediators of lung tissue in each group were evaluated, and the expression of phosphatidylinositol 3-kinase (PI3K)-protein kinase B (AKT)-glycogen synthase kinase (GSK) 3β-nuclear factor erythroid 2-related factor 2 (Nrf2)/nuclear factor (NF)-κB signaling pathway was also detected by using Western blot. Finally, one-way analysis of variance, Welch test, or Kruskal-Wallis H test was used to compare data differences among groups. Results:A total of 415 ingredients were detected from the SSY. 66 of the detected ingredients were identified as active ingredients, and 10 of them were selected as core ingredients. The number of common targets, hub targets, and key hub targets was 337, 50, and 10, respectively. Total of 285 biological processes, 32 cellular components, and 51 molecular functions were enriched though GO analysis, and 148 cell signaling pathways such as pathways in cancer and PI3K-AKT signaling pathway were enriched though KEGG analysis. Molecular docking studies revealed that all binding energies between the 10 key hub targets and the 10 core ingredients were less than -5 kcal/mol. Compared with the ALI group, the levels of the respiratory rate, blood lactate, and lung wet/dry weight ratio in ALI+SSY group were significantly decreased (all P<0.01), and the level of ALI score showed a downward trend, but the difference was not statistically significant ( P>0.05). In addition, the levels of interleukin-6, interleukin-1β, and tumor necrosis factor-α in bronchoalveolar lavage fluid and the levels of malondialdehyde, protein carbonyl, and 8-hydroxy-2-deoxyguanosine in lung tissue of rats in ALI+SSY group were significantly decreased in comparison with those in ALI group (all P<0.01). Moreover, compared with the ALI group, the phosphorylation levels of PI3K p85α, AKT1, and GSK3β and the expression level of Nrf2 in lung tissue of ALI+SSY group were significantly up-regulated (PI3K p85α phosphorylation and AKT1 phosphorylation, P<0.01; GSK3β phosphorylation and Nrf2, P<0.05), while the phosphorylation level of NF-κB p65 was significantly down-regulated ( P<0.01). Conclusions:Active ingredients detected from SSY via untargeted Metabolomics can inhibit oxidative stress and inflammation in ALI rats by regulating the PI3K-AKT-GSK3β-Nrf2/NF-κB signaling pathway, thereby alleviating lung lesions.