Objective:To investigate the diagnostic value and influencing factors of endoscopic ultrasonography (EUS) for detecting rectal neuroendocrine neoplasms (R-NENs).Methods:A retrospective case-control study was performed on data of patients with suspected R-NENs by white light endoscopy who underwent endoscopic diagnosis and treatment or surgical operation and obtained pathological diagnosis at the Affiliated Hospital of Qingdao University from March 2016 to June 2023. Clinical data, EUS characteristics and pathological results were statistically analyzed, and the diagnostic accuracy of EUS for R-NENs were obtained by comparing the EUS results with the pathological results. Influencing factors affecting accuracy were analyzed by using the binary logistic regression model.Results:A total of 317 patients were included. The sensitivity, the specificity, the positive predictive value and the negative predictive value of EUS in diagnosing R-NENs were 98.03% (249/254), 34.92% (22/63), 85.86% (249/290) and 81.48% (22/27) respectively. The accuracy was 85.49% (271/317) and the Jorden index was 0.33. Tumor size ≤5 mm ( P=0.002, OR=2.892, 95% CI: 1.464-5.713), absence of surface vascular dilation ( P=0.019, OR=2.613, 95% CI: 1.170-5.837), normal tumor coloration ( P=0.001, OR=3.460, 95% CI: 1.645-7.279) and erythematous surface appearance ( P=0.048, OR=7.242, 95% CI: 1.015-51.680) were independent risk factors affecting the accuracy of R-NENs diagnosis by EUS. Depth assessment accuracy of EUS was 76.77% (195/254), with echo heterogeneity ( P<0.001, OR=4.008, 95% CI: 1.980-8.113) and surface depression ( P=0.035, OR=2.664, 95% CI: 1.073-6.615) emerging as significant factors affecting invasion depth evaluation. Conclusion:EUS demonstrates substantial clinical utility for R-NENs assessment, with diagnostic performance being significantly associated with tumor morphology and sonographic features. Macroscopic characteristics including tumor size, vascular patterns, and chromatic features influence diagnostic accuracy, while echo-textural heterogeneity and surface depression affect invasion depth precision. These findings underscore the clinical relevance of comprehensive EUS evaluation in R-NENs management.

Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.

Objective:To investigate the diagnostic value and influencing factors of endoscopic ultrasonography (EUS) for detecting rectal neuroendocrine neoplasms (R-NENs).Methods:A retrospective case-control study was performed on data of patients with suspected R-NENs by white light endoscopy who underwent endoscopic diagnosis and treatment or surgical operation and obtained pathological diagnosis at the Affiliated Hospital of Qingdao University from March 2016 to June 2023. Clinical data, EUS characteristics and pathological results were statistically analyzed, and the diagnostic accuracy of EUS for R-NENs were obtained by comparing the EUS results with the pathological results. Influencing factors affecting accuracy were analyzed by using the binary logistic regression model.Results:A total of 317 patients were included. The sensitivity, the specificity, the positive predictive value and the negative predictive value of EUS in diagnosing R-NENs were 98.03% (249/254), 34.92% (22/63), 85.86% (249/290) and 81.48% (22/27) respectively. The accuracy was 85.49% (271/317) and the Jorden index was 0.33. Tumor size ≤5 mm ( P=0.002, OR=2.892, 95% CI: 1.464-5.713), absence of surface vascular dilation ( P=0.019, OR=2.613, 95% CI: 1.170-5.837), normal tumor coloration ( P=0.001, OR=3.460, 95% CI: 1.645-7.279) and erythematous surface appearance ( P=0.048, OR=7.242, 95% CI: 1.015-51.680) were independent risk factors affecting the accuracy of R-NENs diagnosis by EUS. Depth assessment accuracy of EUS was 76.77% (195/254), with echo heterogeneity ( P<0.001, OR=4.008, 95% CI: 1.980-8.113) and surface depression ( P=0.035, OR=2.664, 95% CI: 1.073-6.615) emerging as significant factors affecting invasion depth evaluation. Conclusion:EUS demonstrates substantial clinical utility for R-NENs assessment, with diagnostic performance being significantly associated with tumor morphology and sonographic features. Macroscopic characteristics including tumor size, vascular patterns, and chromatic features influence diagnostic accuracy, while echo-textural heterogeneity and surface depression affect invasion depth precision. These findings underscore the clinical relevance of comprehensive EUS evaluation in R-NENs management.

Objective:To investigate role of miR-4738-3p in targeting tryptophan 2,3-dioxygenase 2(TDO2)to regulate energy metabolism and inhibit immune escape in thyroid cancer cells.Methods:A total of 68 cases of thyroid cancer tissues and their corresponding adjacent non-cancerous tissues were collected,expressions of miR-4738-3p and TDO2 in tissues and cell lines were de-tected by qRT-PCR,relationship between miR-4738-3p expression and clinicopathological characteristics was analyzed.Effects of miR-4738-3p and TDO2 on proliferation,migration,invasion,energy metabolism and immune escape of TPC-1 and BCPAP cells were examined through CCK-8,clone formation,wound healing,Transwell experiments,biochemical analyses,ELISA and NK cell co-culture experiments,targeting relationship between miR-4738-3p and TDO2 was validated by dual luciferase reporter gene assay.In vivo xenograft experiments and immunohistochemical detections were conducted to observe effects of miR-4738-3p on tumorigenesis and expression of immune escape-related proteins in TPC-1 and BCPAP cells.Results:miR-4738-3p was lowly expressed(P<0.05),while TDO2 was highly expressed in thyroid cancer(P<0.05).Tumor size,lymph node metastasis and TNM staging were correlated with expression of miR-4738-3p(P<0.05).Compared with miR-NC group,miR-4738-3p mimics group exhibited decreased prolifera-tion,clone formation,migration,invasion ability,glucose uptake,lactate production,ATP/ADP ratio,TDO2 mRNA and protein expressions,tumor volume,mass,Ki-67,TDO2 and PD-L1 expressions(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were increased(P<0.05).miR-4738-3p inhibitor group showed increased proliferation,clone formation,migration,invasion ability,glucose uptake,lactate production,ATP/ADP ratio,TDO2 mRNA and protein expressions(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were decreased(P<0.05).Binding sites were identified between miR-4738-3p and TDO2,and their expression were negatively correlated(r=-0.485 3,P<0.001).Compared with sh-NC group,proliferation,clone formation,migration,invasion ability,glucose uptake,lactate production and ATP/ADP ratio were decreased in sh-TDO2 group(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were increased(P<0.05).Compared with Scrambled group,proliferation,clone formation,migration,invasion ability,glucose uptake,lactate production and ATP/ADP ratio were increased in TDO2 group(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were decreased(P<0.05).Conclusion:miR-4738-3p targets TDO2 to regulate energy metabolism,inhibiting thyroid cancer cell proliferation,metastasis and immune escape.

Objective:To investigate the effect of ginsenoside octanoate(Rh2-O)on inducing immunogenic cell death in hepa-tocellular carcinoma cells and its molecular mechanism.Methods:Effects of ginsenoside caprylate(Rh2-O)and autophagy inhibitor 3-MA on the activity of hepatocellular carcinoma cells were detected by CCK-8 assay.The effect of Rh2-O on CRT membrane eversion in Hepa1-6 cells were detected by immunofluorescence assay.Rh2-O treated mouse hepatocellular carcinoma cells were used to pre-pare a tumor vaccine for in vivo vaccination experiments in mice.Extracellular ATP levels were detected in real-time.The expression of autophagy-related genes and proteins were measured by real-time fluorescence PCR and Western blot,and the mitochondrial morphol-ogy and co-localization with autophagy proteins were observed by laser confocal microscopy.Results:Rh2-O showed strong cytotoxicity to Hepa1-6 cells[cell viability:(58.54±3.56)%]at a concentration of 150 μmol/L,and a large amount of CRT was observed on the surface of the cell membrane.The tumor emergence rate was 36.36%in the vaccinated group and 100%in the control group.The tumor vaccine prepared by Rh2-O effectively protected mice from the same type of tumor attack;Rh2-O induced an increase in the level of cellular secreted ATP(P<0.05),the mRNA of autophagy-related genes ATG3,p62,LC3 expression levels and autophagy-associated proteins LC3A and LC3B expression levels were increased(P<0.05),and co-localization of mitochondria with autophagy proteins was significantly increased(P<0.05).In addition,Rh2-O action on 3-MA pretreated hepatocellular carcinoma cells resulted in a signifi-cant decrease in extracellular ATP levels(P<0.001).Conclusion:Rh2-O may induce immunogenic cell death by inducing autophagy in hepatocellular carcinoma cells.

Objective:To investigate the effect of ginsenoside octanoate(Rh2-O)on inducing immunogenic cell death in hepa-tocellular carcinoma cells and its molecular mechanism.Methods:Effects of ginsenoside caprylate(Rh2-O)and autophagy inhibitor 3-MA on the activity of hepatocellular carcinoma cells were detected by CCK-8 assay.The effect of Rh2-O on CRT membrane eversion in Hepa1-6 cells were detected by immunofluorescence assay.Rh2-O treated mouse hepatocellular carcinoma cells were used to pre-pare a tumor vaccine for in vivo vaccination experiments in mice.Extracellular ATP levels were detected in real-time.The expression of autophagy-related genes and proteins were measured by real-time fluorescence PCR and Western blot,and the mitochondrial morphol-ogy and co-localization with autophagy proteins were observed by laser confocal microscopy.Results:Rh2-O showed strong cytotoxicity to Hepa1-6 cells[cell viability:(58.54±3.56)%]at a concentration of 150 μmol/L,and a large amount of CRT was observed on the surface of the cell membrane.The tumor emergence rate was 36.36%in the vaccinated group and 100%in the control group.The tumor vaccine prepared by Rh2-O effectively protected mice from the same type of tumor attack;Rh2-O induced an increase in the level of cellular secreted ATP(P<0.05),the mRNA of autophagy-related genes ATG3,p62,LC3 expression levels and autophagy-associated proteins LC3A and LC3B expression levels were increased(P<0.05),and co-localization of mitochondria with autophagy proteins was significantly increased(P<0.05).In addition,Rh2-O action on 3-MA pretreated hepatocellular carcinoma cells resulted in a signifi-cant decrease in extracellular ATP levels(P<0.001).Conclusion:Rh2-O may induce immunogenic cell death by inducing autophagy in hepatocellular carcinoma cells.

Objective:To investigate role of miR-4738-3p in targeting tryptophan 2,3-dioxygenase 2(TDO2)to regulate energy metabolism and inhibit immune escape in thyroid cancer cells.Methods:A total of 68 cases of thyroid cancer tissues and their corresponding adjacent non-cancerous tissues were collected,expressions of miR-4738-3p and TDO2 in tissues and cell lines were de-tected by qRT-PCR,relationship between miR-4738-3p expression and clinicopathological characteristics was analyzed.Effects of miR-4738-3p and TDO2 on proliferation,migration,invasion,energy metabolism and immune escape of TPC-1 and BCPAP cells were examined through CCK-8,clone formation,wound healing,Transwell experiments,biochemical analyses,ELISA and NK cell co-culture experiments,targeting relationship between miR-4738-3p and TDO2 was validated by dual luciferase reporter gene assay.In vivo xenograft experiments and immunohistochemical detections were conducted to observe effects of miR-4738-3p on tumorigenesis and expression of immune escape-related proteins in TPC-1 and BCPAP cells.Results:miR-4738-3p was lowly expressed(P<0.05),while TDO2 was highly expressed in thyroid cancer(P<0.05).Tumor size,lymph node metastasis and TNM staging were correlated with expression of miR-4738-3p(P<0.05).Compared with miR-NC group,miR-4738-3p mimics group exhibited decreased prolifera-tion,clone formation,migration,invasion ability,glucose uptake,lactate production,ATP/ADP ratio,TDO2 mRNA and protein expressions,tumor volume,mass,Ki-67,TDO2 and PD-L1 expressions(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were increased(P<0.05).miR-4738-3p inhibitor group showed increased proliferation,clone formation,migration,invasion ability,glucose uptake,lactate production,ATP/ADP ratio,TDO2 mRNA and protein expressions(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were decreased(P<0.05).Binding sites were identified between miR-4738-3p and TDO2,and their expression were negatively correlated(r=-0.485 3,P<0.001).Compared with sh-NC group,proliferation,clone formation,migration,invasion ability,glucose uptake,lactate production and ATP/ADP ratio were decreased in sh-TDO2 group(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were increased(P<0.05).Compared with Scrambled group,proliferation,clone formation,migration,invasion ability,glucose uptake,lactate production and ATP/ADP ratio were increased in TDO2 group(P<0.05),NAD+/NADH ratio,IFN-γ,TNF-α levels and NK cell killing rate were decreased(P<0.05).Conclusion:miR-4738-3p targets TDO2 to regulate energy metabolism,inhibiting thyroid cancer cell proliferation,metastasis and immune escape.

Objective To investigate the mechanism of action of the nuclear factor erythroid 2-related factor 2(Nrf2)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)signaling pathway in non-alcoholic fatty liver disease(NAFLD),and to explore the mechanism of Gegen QinLian Decoction for the treatment of NAFLD,using in vivo and in vitro experiments.Methods Rats were fed with high-fat chow for 24 weeks to induce NAFLD,and were then divided randomly into normal(C),model(M),high-,medium-,and low-dose Gegen QinLian Decoction(GGQLT-H,GGQLT-M,GGQLT-L),and metformin(Met)groups.From week 25 onwards,the rats were administered the corresponding drugs by gavage for 2 weeks according to the grouping,until sampling.Levels of the oxidative stress markers malondialdehyde(MDA)and glutathione(GSH)in the liver tissues were measured in each group using biochemical kits and ferrous iron(Fe2+)in rat liver tissues was detected using a Fe2+kit.Nrf2,heme oxygenase-1(HO-1),SLC7A11,glutathione synthetase(GSS),GPX4,and acyl coenzyme A synthetase 4(ACSL4)mRNA levels in rat liver tissues were measured by reverse transcription quantitative polymerase chain reaction.For cellular experiments lipid acc umulation was induced in HepG2 hepatocellular carcinoma cells using 1 mmol/L free fatty acid,to mimic the NAFLD in vitro model.Different concentrations of Gegen QinLian Decoction and metformin-containing serum were added for treatment.Lipid accumulation was detected in the cells in each group by Oil red O staining.The MDA and GSH contents of HepG2 cells in the different groups were determined using appropriate kits,and the ferrous contents were detected using a cell-specific ferrous kit.Expression levels of Nrf2,HO-1,SLC7A11,GSS,GPX4,and ACSL4 mRNA was detected in each group of cells using reverse transcription quantitative polymerase chain reaction.Results In the animal experiments,MDA and Fe2+liver levels were significantly higher in the M group than in the C group,while GSH levels were significantly lower(P＜0.01).GGQLT-H,GGQLT-M and Met groups showed significantly reduced MDA and Fe2+and elevated GSH levels compared with the M group(P＜0.01,P＜0.05).High-and medium-dose Gegen QinLian Decoction and metformin increased Nrf2,HO-1,GSS,and GPX4 mRNA and decreased ACSL4 mRNA expression levels(P＜0.01,P＜0.05).In cellular experiments,lipid droplets were significantly increased in the HepG2 cell M group compared with those in the C group,and lipid droplets were significantly reduced by Gegen QinLian Decoction and metformin.MDA and Fe2+levels were significantly increased and GSH levels were significantly decreased in the HepG2 M group compared with the levels in the C group(P＜0.01),while all doses of Gegen QinLian Decoction and metformin significantly decreased MDA and Fe2+levels(P＜0.01)and increased the GSH content(P＜0.01,P＜0.05).Nrf2,GSS,GPX4,and SLC7A11 mRNA expression levels in the GGQLT-H group,Nrf2,HO-1,and SLC7A11 in the GGQLT-L group,HO-1,SLC7A11,and GSS in the GGQLT-M group,and GSS,Nrf2,and HO-1 in the Met group were all significantly increased compared with the findings in the M group(P＜0.01,P＜0.05).ACSL4 mRNA expression levels were significantly decreased in the GGQLT-M and GGQLT-L groups and the Met group(P＜0.01,P＜0.05).Conclusions Gegen QinLian Decoction can improve NAFLD by inhibiting ferroptosis,and its mechanism may he related to regulation of the Nrf2/SLC7A 11/GPX4 signaling pathway.

OBJECTIVE To design and synthesize novel α-naphthylthiol amino acid ester lysine specific demethylase 1(LSD1) inhibitors, evaluate their inhibitory activity with selectivity against LSD1, and explore their binding mechanism through molecular docking and dynamics simulation. METHODS Based on the binding mode of hit compound 3a with LSD1, the α- naphthyl mercapto amino acid ethyl ester small molecule compound were designed by fixing the planar hydrophobic naphthyl ring in the structure, while introducing hydrophilic amino fragment, and they were prepared through a multi-component one-pot cascade reaction. All the compounds were evaluated for their inhibitory activity against LSD1 at concentrations of 5.0 and 1.0 μmol·L–1 using the LSD1 screening platform of research group. The most potent compound was tested for its IC50 value and enzyme selectivity over MAO-A and MAO-B, and its binding mode was investigated through molecular docking and dynamics simulation. RESULTS A total of 13 compounds were obtained, all of which exhibited significant inhibitory effects on LSD1. Among them, nine compounds showed an inhibitory rate of over 50.0% against LSD1 at a concentration of 1.0 μmol·L–1, while compound 3l displaying the best activity with an IC50 value of 0.17 μmol·L–1, 174 times higher than the positive control. It also showed excellent selectivity towards MAO-A and MAO-B. Molecular docking and dynamics simulations indicated that compound 3l inhibited the activity of LSD1 through multiple interactions. CONCLUSION The structures of α-naphthylthiol amino acid ester can serve as lead compounds or active fragments, laying a solid foundation for the subsequent design of LSD1 inhibitors based on structure-oriented drug design.

Objective To visually analyze the application research status quo of artificial intelligence in chronic diseases by using CiteSpace,and to discuss the hot research and evolution process in this field.Methods The SCI-Expanded database of the Web of Science Core Collection(WOSCC)was used as the data source,and the Boolean logic operators"AND,NOT,OR"were used for conducting the subjects terms retriev-al.The application research related information of artificial intelligence in chronic diseases conducted the visu-alized analysis by the CiteSpace software,including the number of published papers,countries,institutions,au-thors,keyword co-occurrence/clustering/emergence and other subjects hotspots,and the scientific map was drawn.Results A total of 1 302 literatures were retrieved and 1 243 literatures meeting the standard were in-cluded in the statistical analysis.Since 2018,the artificial intelligence researches in the field of chronic diseases show a significant growth trend year by year.Among them,the United States published the most,and Harvard University and the University of California were the most prolific institutions.At present,there is a lack of core researchers in this field,moreover the close cooperation among authors is not high.The hot research dis-eases mainly focus on adult heart,lung,kidney,endocrine metabolism and other aspects.Previous researchs fo-cused on intervention therapy,and gradually transitioning to precision medicine,decision support,manage-ment,mental health,mobile health care and other aspects.Conclusion At present,the application research of artificial intelligence in chronic diseases is still a hot research direction,and this paper provides some reference for the research in this field.