ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

BACKGROUND:Spinal cord injury involves mechanisms such as oxidative stress,inflammation,apoptosis and autophagy.Activation of autophagy can improve neuromotor function after spinal cord injury and play a protective role in the spinal cord. OBJECTIVE:To investigate the effects of Periplaneta americana powder on hindlimb motor function and the autophagy protein Beclin-1 in the injured site of rats after spinal cord hemisection. METHODS:Thirty Sprague-Dawley rats,6-8 weeks of age,were randomly divided into three groups(n=10 per group).In the sham-operated group,the lamina was just opened to exposure the spinal cord followed by suturing.Normal saline group and Periplaneta americana powder group both underwent left hemisection of the spinal cord to prepare animal models of spinal cord hemisection.The normal saline group was continuously gavaged with normal saline for 14 days,and the Periplaneta americana powder group was continuously gavaged with Periplaneta americana powder for 14 days.The Basso Beattie Bresnahan scale score was performed at the 6th hour,1st day,3rd day,7th day and 14th day after operation to observe the hindlimb motor function.After 14 days of administration,the rats were sacrificed and sampled.Immunohistochemistry,western blot and immunofluorescence were used to detect the expression of Beclin-1 in the injured site of the spinal cord after hemisection. RESULTS AND CONCLUSION:After operation,the Basso Beattie Bresnahan scale scores were gradually increased in the normal saline group and Periplaneta americana powder group.Compared with the sham-operated group,the Basso Beattie Bresnahan scale scores were significantly reduced in the normal saline group and Periplaneta americana powder group at the 6th hour,1st day,3rd day,7th day and 14th day after operation(P<0.05).The Basso Beattie Bresnahan scale scores in the Periplaneta americana powder group were significantly higher than those in the normal saline group at the 7th and 14th days after operation(P<0.05).Immunohistochemical staining showed that Beclin-1 was weakly positive in the sham-operated group,mainly expressed in the cytoplasm;in the normal saline group,Beclin-1 was mainly expressed in the cytoplasm and partially expressed in the nuclear membrane;in the Periplaneta americana powder group,Beclin-1 was mainly expressed in the cytoplasm and partially expressed in the nuclear membrane.The proportion of Beclin-1 positive cells was higher in the normal saline and Periplaneta americana powder groups than in the sham-operated group(P<0.05),while the proportion of Beclin-1 positive cells was higher in the Periplaneta americana powder group than in the normal saline group(P<0.05).Western blot assay and immunofluorescence staining showed that the Beclin-1 protein expression was higher in the normal saline and Periplaneta americana powder groups than in the sham-operated group(P<0.05),and moreover,the Beclin-1 protein expression was higher in the Periplaneta americana powder group than in the normal saline group(P<0.05).To conclude,Periplaneta americana powder could improve the hindlimb motor function of rats with spinal cord hemisection injury,and the mechanism may be that polysaccharides in the Periplaneta americana powder increase the expression of Beclin-1.

The clinical application of triptolide (TPL) in tumor therapy has been greatly limited by its toxicity and inefficient delivery. Herein, a localized and sustained-release thermo-sensitive hydrogel was developed for the intra-tumor administration of TPL. Based on the amphiphilic structure of poly (

The suitable experimental animal model is important in research of pathogenesis and therapeutic strategies of diabetic foot ulcer, and the murine model is the most commonly used one at present. It can be divided into two types: the animal model simulating pathological conditions and the model simulating clinical symptoms. This article reviews the current research progress on the mechanisms of diabetic ulcer pathogenesis, and relevant treatment strategies, including the inhibition of matrix metalloproteinases (MMPs) expression, promotion of angiogenesis and anti-inflammatory therapy.
Animals ; Anti-Inflammatory Agents ; therapeutic use ; Diabetic Foot ; etiology ; genetics ; therapy ; Disease Models, Animal ; Humans ; Matrix Metalloproteinase Inhibitors ; therapeutic use ; Matrix Metalloproteinases ; genetics ; metabolism ; Mice ; Neovascularization, Physiologic ; physiology