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.

AIM:This study aims to investigate of perodic expression,distribution and interaction between es-trogen receptor α(ERα)and ClC-3 chloride channel,and their relevance to the cell cycle specificity of tamoxifen(TAM)in anti-breast cancer treatment.METHODS:We utilized a web database to analyze the correlation between ERα and ClC-3 expression.Three-dimentional molecular simulation software and co-immunoprecipitation were employed to detect and analyze the interactions between these two proteins.To assess cell cycle dynamics,we performed thymidine(TdR)double-blocking release assay and used nocodazole to block the cell cycle,with subsequent analysis via flow cytometry.Cell viability was measured by MTT assay.Western blot was conducted to evaluate the protein expression levels of ERα and ClC-3,while immunofluorescence staining was utilized to assess their subcellular distribution.RESULTS:(1)Anal-ysis from the web database revealed a significant correlation between ERα and ClC-3 expression,and co-immunoprecipita-tion confirmed their interaction.(2)We successfully obtained human breast cancer T47D cells at different cycle stages us-ing the TdR double-blocking release method and nocodazole treatment.(3)Treatment with TAM primarily inhibited T47D cell viability during G2/M phase.(4)Both ERα and ClC-3 exhibited cyclic variations in protein expression,with their sub-cellular distributions showing periodicity and co-localization.(5)Protein interactions between ERα and ClC-3 were ob-served across all cell cycle phases;(6)After TAM treatment,ERα expression peaked in G2/M phase,while ClC-3 expres-sion remained unaffected.CONCLUSION:Our findings demonstrate cyclic differences in the expression and distribution of ERα and ClC-3 in human breast cancer T47D cells,along with confirmed interactions between these two proteins.The cyclic properties of ERα may play a role in mediating the cell cycle specificity of TAM's anti-breast cancer effect.

Mitochondrial autophagy is a unique mechanism that selectively clears dysfunctional or excess mitochondria,closely related to Alzheimer's disease(AD),which is characterized by aggregated neurotoxic proteins and dysfunctional mitochondria.Numerous recent studies have confirmed that traditional Chinese medicine can have significant therapeutic effects against AD,and its advantages including multi-target,multi-pathway,and multi-action mechanisms have become an important component of AD research.Chinese medicine formulas and monomeric active ingredients,such as Jiannaoyizhi Fang,Danggui Shaoyao Tang,alkaloids,and flavonoids,can regulate mitochondrial autophagy-related signaling pathways and targets,inhibit neuronal mitochondrial autophagy defects,and play a neuroprotective role.This review elaborates on the basic process of mitochondrial autophagy and its related signaling pathways and molecular mechanisms in the pathogenesis of AD,and considers the latest research progress on the use of traditional Chinese medicine to improve AD by regulating mitochondrial autophagy,to provide ideas and references for future basic research and clinical treatment.

Mitochondrial autophagy is a unique mechanism that selectively clears dysfunctional or excess mitochondria,closely related to Alzheimer's disease(AD),which is characterized by aggregated neurotoxic proteins and dysfunctional mitochondria.Numerous recent studies have confirmed that traditional Chinese medicine can have significant therapeutic effects against AD,and its advantages including multi-target,multi-pathway,and multi-action mechanisms have become an important component of AD research.Chinese medicine formulas and monomeric active ingredients,such as Jiannaoyizhi Fang,Danggui Shaoyao Tang,alkaloids,and flavonoids,can regulate mitochondrial autophagy-related signaling pathways and targets,inhibit neuronal mitochondrial autophagy defects,and play a neuroprotective role.This review elaborates on the basic process of mitochondrial autophagy and its related signaling pathways and molecular mechanisms in the pathogenesis of AD,and considers the latest research progress on the use of traditional Chinese medicine to improve AD by regulating mitochondrial autophagy,to provide ideas and references for future basic research and clinical treatment.

Objective To investigate the screening results and factors affecting abnormal detection rates among high-risk groups of esophageal cancer and to explore effective intervention measures. Methods We investigated and collected the information on gender, education level, age, marital status, symptoms of reflux esophagitis (heartburn, acid reflux, belching, hiccup, foreign body sensation in the pharynx, and difficulty swallowing), consumption of pickled vegetables, salt use, and esophageal cancer incidence of villagers in a natural village in Wenfeng District, Anyang City, Henan Province. Changes in reflux esophagitis symptoms in the high-incidence area of esophageal cancer before and after 16 years were observed, and the relationship of such changes with esophageal cancer was analyzed. Results In 2008, 711 cases were epidemiologically investigated, including 213 cases with one of the symptoms of reflux esophagitis such as heartburn, acid reflux, belching, hiccups, foreign body sensation in the pharynx, and difficulty swallowing (sorted in accordance with the abovementioned symptoms; if there are multiple symptoms, then the former is taken); 55 cases with at least two related symptoms, among which the most symptom was heartburn in 108 cases (15.1%); followed by acid reflux, belching, etc. In 2024, the same group of people was investigated, and eight people were missing because of relocation. A total of 703 people were successfully investigated, of which 189 cases (26.8%) had one of the symptoms of reflux esophagitis, such as heartburn (sorted in accordance with relevant symptoms, if there are multiple symptoms, then the former will be selected); 167 cases (23.7%) had at least two related symptoms, among which the most symptom was heartburn in 139 cases (19.7%); followed by acid reflux. Over 16 years, among the 703 people investigated, 45 cases had esophageal cancer and they had one or more of the abovementioned symptoms. No significant differences in the reflux esophagitis-like symptoms were found between 2008 and 2024 (P=0.26); no significant differences in the composition of reflux esophagitis-like symptoms were found between 2008 and 2024 (P=0.299). Conclusion The detection rate of reflux esophagitis-related symptoms in the population in a high-risk area of esophageal cancer has not decreased in the past 16 years, and the high-risk factors still exist. Esophageal cancer is closely related to reflux esophagitis, and clinical practice can provide early diagnosis and treatment for high-risk population.

Objective To understand the disease spectrum of a natural village in an area with high incidence of esophageal cancer to provide a reference for precise prevention and control. Methods From 2008 to 2024, 711 asymptomatic people over the age of 35 years in a natural village with high incidence of esophageal cancer in China were surveyed, and 171 of them were subjected to gastroscopy, biopsy, and pathological examination. All participants were followed up for a long time, and their disease history was recorded. Results A total of 16 years of follow-up were performed, and 703 people were effectively followed up. In 2008, 171 people underwent gastroscopy, and 160 people had biopsy and pathological results in endoscopic screening. By 2024, 76 people had been diagnosed with malignant tumors of 12 different types, and among these people, 45 had esophageal cancer. Conclusion Esophageal cancer remains a significant cause of morbidity and mortality from malignant tumors in this region. Biopsy and pathological examination should be strengthened during gastroscopy, and follow-ups and regular check-ups should be given high importance to reduce the incidence and mortality rates of esophageal cancer.

AIM:This study aims to investigate of perodic expression,distribution and interaction between es-trogen receptor α(ERα)and ClC-3 chloride channel,and their relevance to the cell cycle specificity of tamoxifen(TAM)in anti-breast cancer treatment.METHODS:We utilized a web database to analyze the correlation between ERα and ClC-3 expression.Three-dimentional molecular simulation software and co-immunoprecipitation were employed to detect and analyze the interactions between these two proteins.To assess cell cycle dynamics,we performed thymidine(TdR)double-blocking release assay and used nocodazole to block the cell cycle,with subsequent analysis via flow cytometry.Cell viability was measured by MTT assay.Western blot was conducted to evaluate the protein expression levels of ERα and ClC-3,while immunofluorescence staining was utilized to assess their subcellular distribution.RESULTS:(1)Anal-ysis from the web database revealed a significant correlation between ERα and ClC-3 expression,and co-immunoprecipita-tion confirmed their interaction.(2)We successfully obtained human breast cancer T47D cells at different cycle stages us-ing the TdR double-blocking release method and nocodazole treatment.(3)Treatment with TAM primarily inhibited T47D cell viability during G2/M phase.(4)Both ERα and ClC-3 exhibited cyclic variations in protein expression,with their sub-cellular distributions showing periodicity and co-localization.(5)Protein interactions between ERα and ClC-3 were ob-served across all cell cycle phases;(6)After TAM treatment,ERα expression peaked in G2/M phase,while ClC-3 expres-sion remained unaffected.CONCLUSION:Our findings demonstrate cyclic differences in the expression and distribution of ERα and ClC-3 in human breast cancer T47D cells,along with confirmed interactions between these two proteins.The cyclic properties of ERα may play a role in mediating the cell cycle specificity of TAM's anti-breast cancer effect.

AIM:To investigate the role of ClC-3 chloride channel in the promotion of radio sensitization of na-sopharyngeal carcinoma CNE-2Z cells by dihydroartemisinin(DHA).METHODS:MTT was used to detect the inhibito-ry effect of DHA on the viability of CNE-2Z cells and normal nasopharyngeal epithelial NP69-SV40T cells,the radio sensi-tization effect of DHA on CNE-2Z cells was detected by cloning assay,the expression of ClC-3 protein was detected by Western blot,the expression of ClC-3 protein was down-regulated by siRNA technology,and the chlorine current of cells was recorded by whole cell patch-clamp technology.RESULTS:(1)Compared with NP69-SV40T cells,DHA selective-ly inhibited the proliferation of CNE-2Z cells,with IC10 values of(13.020±4.831)μmol/L and(5.244±1.050)μmol/L,respectively(P＜0.01).(2)The results of clonal formation experiments showed that DHA had a radio sensitizing effect on CNE-2Z cells,with a radio sensitization ratio of 1.9.(3)DHA could activate the chlorine channel of CNE-2Z cells and produce an outward chlorine current,but had no effect on the chlorine channel of NP69-SV40T cells.(4)DHA promoted the expression of ClC-3 chloric channel protein in CNE-2Z cells(P＜0.01).(5)Chlorine channel blocker NPPB could in-hibit the radio sensitizing effect of DHA on CNE-2Z cells by 1.84 times,and also inhibited the chlorine current activated by DHA.(6)the down-regulation of CNE-2Z ClC-3 protein could inhibit the radio sensitization effect of DHA on CNE-2Z cells by 4.19 times,and the activation of chlorine current by DHA on CNE-2Z cells was no longer produced.CONCLU-SION:DHA has a radio sensitizing effect on nasopharyngeal carcinoma CNE-2Z cells,which is likely to be related to the activation of ClC-3 chloride channel.

Objective:To determine the effect of gastrodin(GAS)on toll-like receptor 4(TLR4)expression in mi-croglia after hypoxic-ischemic brain damage.Methods:Hypoxia-ischemic brain damage(HIBD)model was established in neonatal rat in vivo.Thirty 3 d SD rats of were randomly divided into there groups:Sham group,HIBD model group,HIBD model+gastrodin intervention group(HIBD+G).Oxygen glucose deprivation(OGD)model was established in BV2 cells in vitro,Control group(Control),oxygen glucose deprivation group(OGD),OGD+gastrodin intervention group(OGD+G)were randomly set in vitro.Both Western Blot and immunofluorescence staining techniques were used to detect the expression of TLR4 in cells of each group in vitro and in the left corpus callosum region in vivo.Results:The expression of TLR4 was significantly increased in OGD-induced microglia.After gastrodin intervention,TLR4 expression was decreased significantly(P＜0.05).Conclusion:GAS can inhibit the expression of TLR4 in activated microglia and thus play a neuroprotective role in HIBD.

To investigate the expression and significance of cell surface receptor signaling lymphocyte activation molecule family member 7 (SLAMF7) in normal intestinal tissues and intestinal inflammatory tissues of mice.