ObjectiveTo investigate the effect and mechanism of transplantation of human umbilical cord mesenchymal stem cell （hUC-MSC） with overexpression of the Numb gene in the treatment of cholestatic liver fibrosis （CLF）. MethodsThe technique of lentiviral transfection was used to induce the overexpression of the Numb gene in hUC-MSC （hUC-MSC^Numb-OE）， and hUC-MSC transfected with empty vector （hUC-MSC^OE-EV） was used as negative control. Bile duct ligation （BDL） was performed to establish a rat model of CLF， and then the rats were randomly divided into BDL group， hUC-MSC group， hUC-MSC^OE-EV group， and hUC-MSC^Numb-OE group， while a sham-operation group was also established. The rats in the intervention groups were given a single splenic injection of the corresponding cells after BDL， and samples were collected at the end of week 4. Related indicators were measured， including serum biochemistry， liver histopathology， the content of hydroxyproline （Hyp） in the liver， hepatic stellate cell activation， ductular reaction， liver regeneration， and the expression levels of key molecules in the Numb-p53 signaling axis. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the BDL group， the hUC-MSC group and the hUC-MSC^OE-EV group had significant reductions in the levels of serum biochemical parameters （aspartate aminotransferase， gamma-glutamyl transpeptidase， total bile acid， total bilirubin， and direct bilirubin）， liver fibrosis markers （the content of Hyp and the expression levels of alpha-smooth muscle actin， tumor necrosis factor-α， and transforming growth factor-beta 1）， and ductular reaction markers （the expression levels of CK7 and CK19） （all P <0.05）， and compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significantly greater improvements in the above indicators （all P <0.05）. In addition， compared with the hUC-MSC^OE-EV group， the hUC-MSC^Numb-OE group had significant improvements in the expression levels of liver regeneration-related markers （albumin and hepatocyte nuclear factor 4α） and the molecules associated with the Numb-p53 signaling axis （Numb， pNumb， Mdm2， and p53） （all P <0.05）. ConclusionOverexpression of the Numb gene can enhance the therapeutic effect of hUC-MSC on CLF， possibly by activating the Numb-PTB^L-p53-HNF4α axis， promoting the hepatic differentiation of hUC-MSCs and subsequently enhancing liver regeneration.

Mitochondrial biogenesis represents a promising therapeutic target in triple-negative breast cancer (TNBC) due to its essential role in cancer cell metabolism and survival. The natural compound γ-Elemene exhibits potent anti-tumor activity, but its effects on mitochondrial regulation in TNBC remain unclear. In this study, we demonstrate that γ-Elemene induces dose-dependent cytotoxicity in MDA-MB-468 and HCC1806 TNBC cells while significantly impairing mitochondrial function, as shown by reduced membrane potential, oxidative phosphorylation capacity, and ATP production. γ-Elemene treatment markedly suppressed mitochondrial biogenesis, decreasing mitochondrial DNA content and downregulating key mitochondrial genes and proteins. These effects were associated with reduced expression of the master regulators NRF1 and TFAM, but independent of PGC-1α expression levels.Mechanistically, γ-Elemene upregulated the acetyltransferase GCN5, leading to enhanced PGC-1α acetylation. This upregulation occurs primarily through increased GCN5 transcription. Genetic ablation of GCN5 completely reversed γ-Elemene-induced PGC-1α acetylation and restored mitochondrial biogenesis and cell viability, establishing a critical role for GCN5 in mediating these effects. Our findings reveal a novel mechanism whereby γ-Elemene disrupts mitochondrial function in TNBC through GCN5-mediated PGC-1α acetylation, providing new insights into its anti-cancer properties and potential therapeutic applications against TNBC.

Objective To integrate patient clinical information with single-cell sequencing analysis to identify key genes involved in organic erectile dysfunction(ED)in diabetic patients,and to further validate these findings using genome-wide association study(GWAS)data to pinpoint the genes associated with diabetic organic ED.Methods Single-cell RNA sequen-cing data were downloaded from the GSE206528 dataset,comprising samples from five patients including three individuals with-out ED or diabetes,and two patients diagnosed with diabetic ED.Data preprocessing,cell clustering,and annotation were per-formed using R,followed by extraction of microvascular endothelial cells for differential expression analysis.Enrichment analysis of the identified differentially expressed genes(DEGs)was conducted using the Hiplot platform.Expression quantitative trait loci(eQTL)single nucleotide polymorphisms(SNPs)corresponding to these DEGs were retrieved from the UK Biobank(UKB)da-tabase as exposures.ED(GWAS ID:ebi-a-GCST006956)was defined as the outcome variable.Mendelian randomization(MR)analysis was then performed to identify potential causal genes.Results Using the Seurat package,single-cell RNA se-quencing data from the five patients underwent quality control and integration.After cell type identification,a subset of microvas-cular endothelial cells was selected for differential expression analysis,resulting in the identification of 214 DEGs.Functional enrichment analysis revealed that these genes were significantly enriched in pathways related to diabetic complications,including the AGE-RAGE signaling pathway,TNF signaling pathway,and oxidative phosphorylation.Subsequently,MR analysis was per-formed on the 214 DEGs,using erectile dysfunction(ebi-a-GCST006956)as the outcome.Six genes were identified as potential causal genes including MYL9,NFIB,ENDOD1,DES,NRARP and HSPA1B.Conclusion These findings suggest that MYL9,NFIB,ENDOD1,DES,NRARP and HSPA1B may play a role in the progression of organic ED in diabetic patients.

Natural products (NPs) are an important source of new drugs for the treatment of stroke. Identifying cellular targets for bioactive molecules is a major challenge and critical issue in the development of new drugs for stroke. Small-molecule probes play a unique role in target discovery. However, drawbacks to these probes include non-specificity, unstable activity, and difficulty in synthesis. Small-molecule probes based on NPs at least partially compensate for these shortcomings. NPs feature rich chemical and structural diversity, biocompatibility, and unique biological activities. These features could be exploited to provide new ideas and tools for target discovery. Small-molecule probes based on NPs provide a precise and direct search for interacting protein targets of NPs-active small molecules. This review explores the properties of small-molecule probes based on NPs and their applications in mechanistic studies of stroke and other diseases. We hope that this review will bring new perspectives to the mechanistic study of NPs-active small molecules and accelerate the translation of these ingredients into drug candidates for the treatment of stroke.

AIM:This study aims to investigate the mechanism by which Qingfei-Jiedu-Huatan Formula(QJHF)regulates autophagy in alveolar macrophages through mTOR in the treatment of severe pneumonia(SP)in rats.METHODS:Sixty SPF-grade male rats were randomly assigned to six groups:control,model,QJHF,moxifloxacin(MOX),rapamycin(RAPA),and QJHF+RAPA,with ten rats in each group.An SP rat model was established using Klebsiella pneumoniae.After seven days of treatment,changes in IL-33 and IFN-γ levels in bronchoalveolar lavage fluid(BALF)were measured using ELISA.Histopathological alterations in lung tissue were assessed via HE staining,and the autophagy of alveolar macrophages was detected using immunofluorescence co-localization methods.The expression levels of mTOR,beclin-1,and LC3 mRNA in lung tissue were analyzed using qPCR,while Western blot was employed to assess the protein levels of p-mTOR/mTOR,beclin-1,and LC3-II/LC3-I.RESULTS:Compared to the control group,the model group exhibited a deteriorated condition,characterized by alveolar wall rupture and thickening,significant inflammatory cell infiltration in the alveolar cavity,and extensive lung tissue damage(P<0.01).Elevated levels of IL-33 and IFN-γ in BALF were also observed(P<0.01),along with increased colocalization of CD68 and LC3 in immunofluorescence analy-sis.The mTOR mRNA expression in lung tissue decreased(P<0.01),while LC3 and beclin-1 mRNA expressions in-creased(P<0.01).Additionally,the protein expression ratio of p-mTOR/mTOR decreased(P<0.01),whereas LC3-II/LC3-I and beclin-1 protein levels increased(P<0.01).In comparison to the model group,significant improvements were noted after treatment with QJHF and MOX(P<0.01),while RAPA treatment led to a worsening of these indicators(P<0.05).A slight improvement was observed with the QJHF combined with RAPA intervention,though this was not statisti-cally significant.No significant differences were found between the MOX and QJHF groups.However,the QJHF+RAPA group displayed notable improvements in various indicators compared to the RAPA group(P<0.05).CONCLUSION:The QJHF can mitigate the inflammatory response associated with severe pneumonia,potentially by activating mTOR phos-phorylation activity,which in turn inhibits excessive autophagy in alveolar macrophages.

Objective: To explore the expression of collagen type 1 alpha 2 (COL1A2) in cervical cancer and its correlation with immune infiltration. Methods: Bioinformatics techniques were used to analyze the expression of COL1A2 in cervical cancer. Western blot and RT-qPCR were used to detect the expression of COL1A2 in cervical cancer tissues and cell lines. The correlation between the expression of COL1A2 and tumor immune cell infiltration was analyzed by tumor immune estimation resource (TIMER2. 0) . Gene set enrichment analysis (GSEA) was used to analyze the possible mechanism of COL1A2 in cervical cancer. Jaspar database was used to predict the transcrip- tion factors of COL1A2. Western blot and RT-qPCR were used to detect the expression of transcription factors in cervical cancer tissues and cell lines. Results: The expression of COL1A2 was down-regulated in cervical cancer (P < 0. 05) . The expression of COL1A2 was positively correlated with the levels of macrophages and myeloid den- dritic cells (P < 0. 01) . The proportions of 22 types of immune cells were different in different cervical cancer pa- tients. In addition , compared with the high expression group of COL1A2 , the proportion of M0 macrophages , M2 macrophages and resting memory CD4 + T cells increased in the low expression group of COL1A2 , while the propor- tion of CD8 + T cells , activated memory CD4 + T cells , follicular helper T cells , activated NK cells and activated myeloid dendritic cells decreased (P < 0. 05) . GSEA analysis showed that COL1A2 was related to immune-related signaling pathways , including Notch signaling pathway , interleukin-6/janus kinase/signal transducer and activator of transcription 3 (IL6/JAK/STAT3) , Wnt/β-catenin signaling pathway , etc. (P < 0. 01) . Jaspar database pre- dicted that the transcription factor of COL1A2 was paired box protein 5 (PAX5) , and the expression of PAX5 de- creased in cervical cancer (P < 0. 05) . Conclusion COL1A2 is expected to become a potential diagnostic biomar- ker and immunotherapy target for cervical cancer.

Objective:To explore the efficacy and safety of the combination therapy of lenalidomide and rituximab (R2) for short-cycle maintenance therapy in patients with B-cell non-Hodgkin lymphoma (B-NHL).Methods:A retrospective case series study was conducted. The clinical data of 19 B-NHL patients who received R2 regimen maintenance therapy after achieving complete remission through chemotherapy or autologous hematopoietic stem cell transplantation at Dongguan Kanghua Hospital from February 2018 to January 2024 were collected, and the overall survival (OS), progression-free survival (PFS), adverse reactions, changes in lymphocyte subsets and cytokine levels before and after treatment were analyzed.Results:Among the 19 patients, there were 7 males and 12 females, with a median age [ M ( Q1, Q3)] of 49 (45, 65) years. The median follow-up time was 56 months, ranging from 5 to 77 months. The 1-year OS and PFS rates were 89.2% and 88.9%, respectively. The 2-year and 5-year PFS rates were both 83.2%, and the 2-year and 5-year OS rates were both 88.9%. Common adverse reactions included hematological adverse reactions and infections, with 4 cases (21.1%) experiencing grade 3-4 hematological adverse reactions and 4 cases (21.1%) experiencing infections. There was no statistically significant difference in the levels of lymphocyte subsets (total T cells, helper T cells, regulatory T cells, NK cells, B cells, and CD4/CD8) and cytokines [interleukin (IL)-2, IL-4, IL-6, IL-10, tumor necrosis factor-α, and interferon-γ] before and after treatment (all P > 0.05). Conclusions:The R2 regimen for short-cycle maintenance therapy of B-NHL is effective and well tolerated by patients.

Objective:To explore the efficacy and safety of instantly generated inhaled nitric oxide (iNO) for treating neonatal pulmonary hypertension (PH) complicated with severe hypoxic respiratory failure.Methods:This single-center, single-arm, prospective study included 32 neonates with PH complicated with hypoxic respiratory failure who were hospitalized in the neonatal intensive care unit (NICU) of Beijing Children′s Hospital Affiliated to Capital Medical University from March 2023 to March 2025 and received immediate iNO generation therapy. The demographic data, maternal pregnancy, mechanical ventilation parameters, arterial blood gas indexes, other hospitalization data and safety indexes of iNO treatment were collected. The time point for starting iNO treatment was set as 0 h, and the observation time points were 1, 6, 12, 24, 48 h after treatment and when iNO treatment was stopped. The positive reaction of iNO treatment was defined as the decrease of oxygenation index (OI)>10% or the increase of arterial partial pressure of oxygen (PaO 2)>10% after treatment. The OI, mechanical ventilation parameters, arterial blood gas index changes and treatment positive reaction ratio were analyzed to evaluate the effectiveness of iNO treatment, and the nitrogen dioxide concentration, methemoglobin (MetHb) concentration and other indicators were analyzed to evaluate the safety of iNO treatment. Paired t test or Wilcoxon signed rank sum test was used to compare the observation indexes at different treatment times. Friedman test was used to compare the concentration of nitrogen dioxide and MetHb at multiple treatment times. Receiver operating characteristic (ROC) curve was used to analyze the best cut-off value of OI related indexes to distinguish the treatment outcome of iNO. Results:Among 32 neonates, 18 (56%) were males and 14 (44%) were females, the gestational age was 38 (35, 39) weeks, the birth weight was 3.1 (2.3, 3.4) kg, and the age of enrollment was 3 (2, 8) days. The OI and the mean airway pressure at 48 h after treatment were both lower than those at 0 h ((10.4±2.0 vs. 22.6±2.5, 13.0 (12.0, 14.0) vs. 14.0 (13.0, 16.0) cmH 2O, 1 cmH 2O=0.098 kPa, both P<0.05). The fraction of inspired oxygen at 24 and 48 h after treatment were both lower than those at 0 h (both P<0.05). The PaO 2 at 6, 12, 24 and 48 h after treatment were all higher than those at 0 h (all P<0.05). The proportion of positive reactions to iNO treatment was 20 neonates (63%), 22 neonates (69%), 23 neonates (72%), 23 neonates (72%) and 26 neonates (8%) at 1, 6, 12, 24, 48 h after treatment, respectively. No occurrence of methemoglobinemia, excessive nitrogen dioxide concentration, or device related adverse events were observed. Out of 32 neonates, a total of 24 neonates (75%) were cured or improved and discharged according to medical advice, while 8 neonates (25%) died in the hospital. The best cut-off value of OI at 0 h and the decline range of OI at 12 h to distinguish the outcome of hospitalization were 24.8 and 22.2%, respectively. Conclusion:It was effective and safe to use instantly generated iNO to treat neonatal PH with severe hypoxic respiratory failure.

Objective:To investigate how gut microbiota changes during prostate cancer radiotherapy and decipher the relationship of gut microbiota with disease progression and chronic radiation enteritis.Methods:Thirty-one patients with prostate cancer were included in this study,admitted to The First Affiliated Hospital of Xinjiang Medical University from September 2022 to December 2023.The clinical data and stool samples of the patients were collected,and patients were followed up.The collected stool specimens were subjected to 16S rRNA se-quencing to detect gut microbiota and bioinformatics analysis.Results:The relative abundance of phyla such as Firmicutes and Actinobac-teria increased,and that of Bacteroidetes decreased(P<0.05)with an increasing radiotherapeutic dose,while beta diversity was significantly higher(P=0.001).The relative abundance of the phylum Actinobacteria was significantly higher in the prostate cancer progression group than in the non-progression group(P<0.05),the relative abundances of genera such as Sutterella and Haemophilus were significantly higher in the progression group(P<0.05).That of Verrucomicrobia and its offshoots in Akkermansia was higher in the chronic radiation enteritis than in the non-enteritis group(P<0.05),while the relative abundances of Coprococcus_1 and Catabacter in the non-enteritis group were higher than those in the enteritis group(P<0.05).Conclusions:Radiotherapy dose accumulation significantly remodeled the floral structure.Sutterella and Haemophilus of the phylum Proteobacteria might be key flora in prostate cancer recurring early after treatment.An augmen-ted abundance of Akkermansia might increase the risk of chronic radiation enteritis,whereas the flora under the Lachnospiraceae branch might exert aprotective effect against chronic radiation enteritis.

Objective:To revise and enlarge the specification of pharmaceutical excipient sucrose palmitate.Methods:Reference to JP2018,USP-NF 2023,EP1 1.0,Chinese Pharmacopoeia 2020 edition of the four general rules and the first supplement of sucrose stearate pharmacopoeia standards and other relevant requirements for standard research.Results:According to the quality of the product and the actual application in the formulation,the quality standards for the pharmaceutical excipient sucrose palmitate will be studied.At present,the quality standard of sucrose palmitate for pharmaceutical excipients has been disclosed.Conclusion:The established stand-ard will provide the quality guarantee for the application of sucrose palmitate in medicines.