Objective: To investigate the regulatory role of Porphyromonas gingivalis(Pg) infection on the EGFR/GSK3β signaling axis, and its impact on epithelial-mesenchymal transition(EMT) and cetuximab(Ctx) resistance in esophageal squamous cell carcinoma(ESCC). Methods: Single cell RNA sequencing was employed to perform differential analysis of cellular subpopulations, identifying differentially expressed genes in ESCC tissues infected and non-infected with Pg. IHC was conducted to assess the expression of Pg and epidermal growth factor receptor(EGFR) in ESCC tissues. Western blot, RT-PCR, and IF staining were performed to evaluate EGFR expression in Pg infected ESCC cell lines KYSE70 and TE1. ESCC cells were treated with Pg and EGFR inhibitor Ctx, and divided into four groups: control(NC) group, Pg group, Ctx group, Pg+Ctx group. Cell proliferation, migration and invasion abilities were evaluated using CCK-8, plate cloning, wound healing and Transwell assay. Western blot analysis was performed to detect the expression of EMT and EGFR/GSK3β signaling pathway-associated proteins and their phosphorylation levels. Transforming growth factor-β1(TGF-β1) was used to induce EMT in ESCC cells, promoting a transition from the epithelial phenotype to mesenchymal-like phenotype. The differential effects of Ctx on these two phenotypic states were subsequently compared. Results: Epithelial cells were predominantly enriched in Pg-positive tissues, and Pg infection promoted the upregulation of EGFR expression in ESCC cells. Compared to the NC group, Pg treatment significantly enhanced the proliferation, invasion and migration capabili-ties of ESCC cells, and also increased chemoresistance to Ctx and reduced its antitumor efficacy. Pg induced EMT in ESCC cellsviathe EGFR/GSK3β signaling pathway. Notably, Ctx exhibited markedly weaker inhibitory effects on mesenchymal-like cells compared to epithelial ESCC cells. Conclusion Pg promotes ESCC cells proliferation, invasion and migration by regulating EMT through the EGFR/GSK3β signaling pathway, and enhances chemoresistance to Ctx.

Attention is the cornerstone of effective functioning in a complex and information-rich world. While the neural activity of attention has been extensively studied in the cortex, the brain-wide neural activity patterns are largely unknown. In this study, we conducted a comprehensive analysis of neural activity across the mouse brain during attentional processing using EEG and c-Fos staining, utilizing hierarchical clustering and c-Fos-based functional network analysis to evaluate the c-Fos activation patterns. Our findings reveal that a wide range of brain regions are activated, notably in the high-order cortex, thalamus, and brain stem regions involved in advanced cognition and arousal regulation, with the central lateral nucleus of the thalamus as a strong hub, suggesting the crucial role of the thalamus in attention control. These results provide valuable insights into the neural network mechanisms underlying attention, offering a foundation for formulating functional hypotheses and conducting circuit-level testing.
Animals ; Attention/physiology* ; Mice ; Brain/physiology* ; Male ; Electroencephalography ; Reaction Time/physiology* ; Brain Mapping ; Mice, Inbred C57BL ; Choice Behavior/physiology* ; Proto-Oncogene Proteins c-fos/metabolism*

Objective To explore the mechanism of electroacupuncture in alleviating hyperlipidemia in a rat model by modulating mammalian target of rapamycin complex 1(mTORC1)-mediated lipophagy in hepatocytes.Methods A total of 30 SD rats were randomly divided into blank(n=6)and modeling groups(n=24)using the random number table method.A hyperlipidemic rat model was established by feeding rats a high-fat diet(feeding for 8 weeks).After successful modeling,the modeling group was randomly divided into the model,electroacupuncture,mTORC1 inhibitor,and electroacupuncture+mTORC1 agonist groups,with six rats in each group.Except for the blank group,all other rats were fed with high fat diet.Rats in the electroacupuncture and electroacupuncture+mTORC1 agonist groups received electroacupuncture intervention at bilateral"Fenglong"(ST40)acupoints(dilatational wave 2 Hz/100 Hz,current intensity 1 mA)for 30 min once daily.Rats in the mTORC1 inhibitor group received intraperitoneal injections of the mTORC1 inhibitor,rapamycin(2 mg/kg),once daily.Rats in the electroacupuncture+mTORC1 agonist group received intraperitoneal injections of the mTORC1 agonist MHY1485(10 mg/kg)once daily.The interventions were administered for five consecutive days per week for 4 weeks.Upon completion of the intervention,the following analyses were performed:serum contents of total cholesterol(TC),triglycerides(TAG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),free fatty acids(FFA),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were measured using a fully automated biochemical analyzer.Hepatic histopathological changes and lipid deposition were observed using hematoxylin-eosin and oil red O staining.The liver condition was observed and the liver index was calculated.Hepatic TC and TAG levels were measured using an enzyme-linked immunosorbent assay.The ultrastructure of the liver tissue was observed using transmission electron microscopy,and the mean fluorescence intensity of perilipin 2(PLIN2)and microtubule-associated protein 1 light chain 3(LC3)-Ⅱ in the liver tissue was detected using immunofluorescence.Protein expression of LC3-Ⅱ/LC3-Ⅰ,phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR,and mTORC1 in liver tissue was detected using Western blotting.Results Compared to the blank group,the model group rats showed increased serum TC,TAG,LDL-C,ALT,AST,and FFA levels,along with decreased HDL-C levels(P<0.05).The liver index and hepatic TC and TAG levels were also elevated(P<0.05).Histological examination of liver tissue revealed substantial lipid accumulation,numerous lipid droplets within hepatocytes,abnormal mitochondrial morphology,and scarce autophagic vacuole.The mean fluorescence intensity of PLIN2 increased,whereas that of LC3-Ⅱ decreased(P<0.05).Additionally,the LC3-Ⅱ/LC3-Ⅰ ratio was reduced,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression were increased(P<0.05).Compared to the model group,rats in the mTORC1 inhibitor and electroacupuncture groups exhibited decreased serum TC,TAG,LDL-C,ALT,AST,and FFA levels(P<0.05),along with a reduced liver index and hepatic TC and TAG levels(P<0.05).Histological examination showed markedly attenuated lipid accumulation and visible autophagic vacuole in the hepatocytes.The mean fluorescence intensity of PLIN2 decreased,whereas that of LC3-Ⅱ increased(P<0.05).Moreover,the LC3-Ⅱ/LC3-Ⅰ ratio increased,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression decreased(P<0.05).In comparison with both the electroacupuncture and mTORC1 inhibitor groups,the electroacupuncture+mTORC1 agonist group demonstrated increased serum TAG,TC,LDL-C,ALT,AST,and FFA levels(P<0.05)as well as elevated liver index and hepatic TC and TAG levels(P<0.05).Liver tissues exhibited aggravated lipid deposition and absence of autophagic vacuole in liver cells.The mean fluorescence intensity of PLIN2 was enhanced,whereas that of LC3-Ⅱ was reduced(P<0.05).Furthermore,the LC3-Ⅱ/LC3-Ⅰ ratio decreased,and the p-mTOR/mTOR ratio and mTORC1 protein expression increased(P<0.05).Conclusion Electroacupuncture at"Fenglong"(ST40)may improve blood lipid levels in hyperlipidemic rats by inhibiting mTORC1 and promoting hepatocyte lipophagy.

Objective:To investigate the impact and clinical outcomes of intraoperative prone-position lumbar anteroposterior (AP) fluoroscopy under anesthesia on the selection of the lowest instrumented vertebra (LIV) in the patients with adolescent idiopathic scoliosis (AIS) plus structural lumbar curves.Methods:A retrospective analysis was conducted of the clinical data of 35 patients (29 females and 6 males) with AIS who had undergone surgical posterior correction and fusion at Scoliosis Center, The Third Affiliated Hospital, Sun Yat-sen University between January 2020 and October 2023. The mean age was (17.9±5.7) years. Lenke's classification: 6 cases of type 3, 12 cases of type 4, 7 cases of type 5 and 10 cases of type 6. Preoperatively, all patients underwent standing AP and lateral radiographs of the full-length spine, left and right bending radiographs of the spine, and full-spine CT. Intraoperatively, all patients underwent prone-position lumbar AP fluoroscopy under anesthesia. The criteria for LIV selection were: (1) it should be the most cephalad vertebra touched by the central sacral vertical line (CSVL); (2) its rotation should be ≤ grade Ⅱ by the Nash-Moe classification; (3) its tilt angle should be <25°. The preoperative and postoperative LIV rotation angles were compared, and the number of lumbar fusions was compared between preoperative planning and actual surgery. Comparisons were also made between preoperation, postoperation and the final follow-up, examining Cobb angle of the major curve, Cobb angle of the minor curve, LIV inclination, coronal balance distance (CBD), sagittal vertical axis (SVA), and distance between CSVL and LIV (CSVL-LIV). The correction rates of the major curve and the minor curve, and change in LIV inclination were compared between postoperation and the final follow-up.Results:The patients were followed for (18.0±3.0) months. The LIV rotation decreased from 8.34°±4.95° preoperatively to 5.03°±2.99° postoperatively. The intraoperative fluoroscopy reduced at least one segment fusion for 57.1% (20/35) of the patients so that the number of lumbar fusions decreased significantly from 4.2±0.7 in preoperative planning to 3.6±0.8 after actual surgery ( P<0.05). The Cobb angles of the major and minor curves, LIV inclination, and CSVL-LIV at postoperation and the final follow-up were significantly lower than the preoperative values ( P<0.05), but there were no significant differences between the final follow-up and postoperation in the Cobb angle of the major cure, Cobb angle of the minor curve, or LIV inclination ( P>0.05). None of the patients required surgical revision for distal junctional complications. Conclusions:In the surgical treatment of AIS patients with structural lumbar curves, compared to the preoperative X-rays using the same criteria, intraoperative prone-position lumbar AP fluoroscopy under anesthesia can not only be a safe and effective method for LIV selection but also effectively reduce the number of lumbar fusions to preserve more lumbar mobility.

Objective:To investigate the impact and clinical outcomes of intraoperative prone-position lumbar anteroposterior (AP) fluoroscopy under anesthesia on the selection of the lowest instrumented vertebra (LIV) in the patients with adolescent idiopathic scoliosis (AIS) plus structural lumbar curves.Methods:A retrospective analysis was conducted of the clinical data of 35 patients (29 females and 6 males) with AIS who had undergone surgical posterior correction and fusion at Scoliosis Center, The Third Affiliated Hospital, Sun Yat-sen University between January 2020 and October 2023. The mean age was (17.9±5.7) years. Lenke's classification: 6 cases of type 3, 12 cases of type 4, 7 cases of type 5 and 10 cases of type 6. Preoperatively, all patients underwent standing AP and lateral radiographs of the full-length spine, left and right bending radiographs of the spine, and full-spine CT. Intraoperatively, all patients underwent prone-position lumbar AP fluoroscopy under anesthesia. The criteria for LIV selection were: (1) it should be the most cephalad vertebra touched by the central sacral vertical line (CSVL); (2) its rotation should be ≤ grade Ⅱ by the Nash-Moe classification; (3) its tilt angle should be <25°. The preoperative and postoperative LIV rotation angles were compared, and the number of lumbar fusions was compared between preoperative planning and actual surgery. Comparisons were also made between preoperation, postoperation and the final follow-up, examining Cobb angle of the major curve, Cobb angle of the minor curve, LIV inclination, coronal balance distance (CBD), sagittal vertical axis (SVA), and distance between CSVL and LIV (CSVL-LIV). The correction rates of the major curve and the minor curve, and change in LIV inclination were compared between postoperation and the final follow-up.Results:The patients were followed for (18.0±3.0) months. The LIV rotation decreased from 8.34°±4.95° preoperatively to 5.03°±2.99° postoperatively. The intraoperative fluoroscopy reduced at least one segment fusion for 57.1% (20/35) of the patients so that the number of lumbar fusions decreased significantly from 4.2±0.7 in preoperative planning to 3.6±0.8 after actual surgery ( P<0.05). The Cobb angles of the major and minor curves, LIV inclination, and CSVL-LIV at postoperation and the final follow-up were significantly lower than the preoperative values ( P<0.05), but there were no significant differences between the final follow-up and postoperation in the Cobb angle of the major cure, Cobb angle of the minor curve, or LIV inclination ( P>0.05). None of the patients required surgical revision for distal junctional complications. Conclusions:In the surgical treatment of AIS patients with structural lumbar curves, compared to the preoperative X-rays using the same criteria, intraoperative prone-position lumbar AP fluoroscopy under anesthesia can not only be a safe and effective method for LIV selection but also effectively reduce the number of lumbar fusions to preserve more lumbar mobility.

Insulinoma-associated protein 1(INSM1)is an emerging neuroendocrine marker that has received ex-tensive attention in previous studies.INSM1 demonstrates high sensitivity and specificity in neuroendocrine neoplasms(NENs)across various sites and grades.However,some studies have pointed out some inconsistencies in its expres-sion.This article explores the specific expression patterns of INSM1 in NENs of different systems,as well as in other tumors that express neuroendocrine markers.It also analyzes the critical role of INSM1 in the diagnosis of neuroendo-crine tumors and reviews its application value,prospects,and problems.

Objective To explore the mechanism of electroacupuncture in alleviating hyperlipidemia in a rat model by modulating mammalian target of rapamycin complex 1(mTORC1)-mediated lipophagy in hepatocytes.Methods A total of 30 SD rats were randomly divided into blank(n=6)and modeling groups(n=24)using the random number table method.A hyperlipidemic rat model was established by feeding rats a high-fat diet(feeding for 8 weeks).After successful modeling,the modeling group was randomly divided into the model,electroacupuncture,mTORC1 inhibitor,and electroacupuncture+mTORC1 agonist groups,with six rats in each group.Except for the blank group,all other rats were fed with high fat diet.Rats in the electroacupuncture and electroacupuncture+mTORC1 agonist groups received electroacupuncture intervention at bilateral"Fenglong"(ST40)acupoints(dilatational wave 2 Hz/100 Hz,current intensity 1 mA)for 30 min once daily.Rats in the mTORC1 inhibitor group received intraperitoneal injections of the mTORC1 inhibitor,rapamycin(2 mg/kg),once daily.Rats in the electroacupuncture+mTORC1 agonist group received intraperitoneal injections of the mTORC1 agonist MHY1485(10 mg/kg)once daily.The interventions were administered for five consecutive days per week for 4 weeks.Upon completion of the intervention,the following analyses were performed:serum contents of total cholesterol(TC),triglycerides(TAG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),free fatty acids(FFA),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were measured using a fully automated biochemical analyzer.Hepatic histopathological changes and lipid deposition were observed using hematoxylin-eosin and oil red O staining.The liver condition was observed and the liver index was calculated.Hepatic TC and TAG levels were measured using an enzyme-linked immunosorbent assay.The ultrastructure of the liver tissue was observed using transmission electron microscopy,and the mean fluorescence intensity of perilipin 2(PLIN2)and microtubule-associated protein 1 light chain 3(LC3)-Ⅱ in the liver tissue was detected using immunofluorescence.Protein expression of LC3-Ⅱ/LC3-Ⅰ,phosphorylated mammalian target of rapamycin(p-mTOR)/mTOR,and mTORC1 in liver tissue was detected using Western blotting.Results Compared to the blank group,the model group rats showed increased serum TC,TAG,LDL-C,ALT,AST,and FFA levels,along with decreased HDL-C levels(P<0.05).The liver index and hepatic TC and TAG levels were also elevated(P<0.05).Histological examination of liver tissue revealed substantial lipid accumulation,numerous lipid droplets within hepatocytes,abnormal mitochondrial morphology,and scarce autophagic vacuole.The mean fluorescence intensity of PLIN2 increased,whereas that of LC3-Ⅱ decreased(P<0.05).Additionally,the LC3-Ⅱ/LC3-Ⅰ ratio was reduced,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression were increased(P<0.05).Compared to the model group,rats in the mTORC1 inhibitor and electroacupuncture groups exhibited decreased serum TC,TAG,LDL-C,ALT,AST,and FFA levels(P<0.05),along with a reduced liver index and hepatic TC and TAG levels(P<0.05).Histological examination showed markedly attenuated lipid accumulation and visible autophagic vacuole in the hepatocytes.The mean fluorescence intensity of PLIN2 decreased,whereas that of LC3-Ⅱ increased(P<0.05).Moreover,the LC3-Ⅱ/LC3-Ⅰ ratio increased,whereas the p-mTOR/mTOR ratio and mTORC1 protein expression decreased(P<0.05).In comparison with both the electroacupuncture and mTORC1 inhibitor groups,the electroacupuncture+mTORC1 agonist group demonstrated increased serum TAG,TC,LDL-C,ALT,AST,and FFA levels(P<0.05)as well as elevated liver index and hepatic TC and TAG levels(P<0.05).Liver tissues exhibited aggravated lipid deposition and absence of autophagic vacuole in liver cells.The mean fluorescence intensity of PLIN2 was enhanced,whereas that of LC3-Ⅱ was reduced(P<0.05).Furthermore,the LC3-Ⅱ/LC3-Ⅰ ratio decreased,and the p-mTOR/mTOR ratio and mTORC1 protein expression increased(P<0.05).Conclusion Electroacupuncture at"Fenglong"(ST40)may improve blood lipid levels in hyperlipidemic rats by inhibiting mTORC1 and promoting hepatocyte lipophagy.

Insulinoma-associated protein 1(INSM1)is an emerging neuroendocrine marker that has received ex-tensive attention in previous studies.INSM1 demonstrates high sensitivity and specificity in neuroendocrine neoplasms(NENs)across various sites and grades.However,some studies have pointed out some inconsistencies in its expres-sion.This article explores the specific expression patterns of INSM1 in NENs of different systems,as well as in other tumors that express neuroendocrine markers.It also analyzes the critical role of INSM1 in the diagnosis of neuroendo-crine tumors and reviews its application value,prospects,and problems.

Objective:To describe the epidemiological distribution of hemorrhoids in a physical exami-nation population in China,which could provide evidence for precision prevention and early intervention of hemorrhoids.Methods:Chinese subjects over 18 years of age who underwent a physical examination in a nationwide chain of physical examination centers in 2018 were studied in a cross-sectional design,which collected information by a questionnaire and physical examination results from each subject.The epidemiological distribution of hemorrhoids was described using Logistic models.The gender-,age-,and region-detection rates of hemorrhoids were standardized to the Sixth National Population Census of the People's Republic of China(2010).Results:A total of 2 940 295 adult subjects were included in the study,of whom the average age was(41.7±14.0)years,and 52.6％were females.The standardized detection rate of hemorrhoids was higher for females(43.7％)than that for males(17.7％;P＜0.001)in this study.In the females,the age distribution of hemorrhoids was inverted U-shaped,with the highest standardized detection rate of hemorrhoids in the age group of 30-39 years(63.5％).In the males,the standardized detection rate of hemorrhoids increased along with age,with the highest percentage of 17.2％in the age group of 50-59 years,and the standardized detection rate of hemorrhoids in the age group of 60 and above decreased slightly(P＜0.001 for trend test).The participants with hypertension had a higher standardized detection rate of hemorrhoids than those with normal blood pressure in both males and females(P＜0.001).The standardized detection rate of hemorrhoids showed a positive corre-lation with body mass index(P＜0.001 for trend test in males).Conclusion:The detection rate of hemorrhoids varied to gender,age,obesity,and hypertension status,which could help to identify the risk factors and the high-risk sub-groups,and hence to strengthen health education and early detection accordingly,which could eventually reduce the incidence of hemorrhoids and improve the quality of life and health in the Chinese population.This study was conducted in a physical examination population,and the conclusions of this study should be extrapolated with caution.

Hormonal receptor positive human epidermal receptor 2 negative (HR⁺/HER2^-) is the commonest molecular subtype of breast cancer (BC). Patients with HR⁺/HER2^- BC may manifest clinically a late recurrence whose BC metastasizes 10-15 years post-operatively. We report one case who presented with pulmonary mass in upper lobe of lung and Horner syndrome 16 years after BC surgery. FDG PET/CT suggested pulmonary malignancy but could not differentiate between primary or metastatic cancer when invasive biopsy was quite risky. Novel ¹⁸F-FES PET/CT facilitated the non-invasive functional diagnosis of estrogen-receptor positive (ER+) pulmonary metastasis of BC, and the patient experienced partial response (PR) after CDK4/6 inhibitor and aromatase inhibitor as endocrine therapy. This article reviews the diagnosis and treatment process of this case, to provide guidance for non-invasive global evaluation of ER status among metastatic HR⁺/HER2^- BC patients with ¹⁸F-FES PET/CT.