1.The Clinical and Genetics Characteristics of Oculopharyngodistal Myopathy
Jiaxi YU ; Zhihao QUAN ; Yilei ZHENG ; Jing AN ; Jing LIU ; Qingqing WANG ; Lingchao MENG ; Meng YU ; Zhiying XIE ; Jianwen DENG ; He LYU ; Wei ZHANG ; Yun YUAN ; Zhaoxia WANG
JOURNAL OF RARE DISEASES 2026;5(2):164-174
To analyze the clinical and genetic features of oculopharyngodistal myopathy (OPDM) patients and compare the phenotypic differences among various causative genes. A total of 65 genetically confirmed OPDM patients from 43 unrelated families, who were admitted to the Department of Neurology, Peking University First Hospital between January 2008 and December 2025, were retrospectively included.The general demographic data, clinical manifestations, laboratory/auxiliary examinations, muscle pathology, and genetic test results were systematically collected and analyzed. The clinical and pathological characteristics among different OPDM subtypes were compared. Among the 65 patients(39 male and 26 female), the mean age of onset was (31.20±10.43) years (range: 14 to 63 years). The initial symptom was predominantly distal limb weakness (67.44%), which gradually progressed to involve the extraocular muscles, pharyngeal muscles, facial muscles and proximal limb muscles. Serum creatine kinase levels were mildly to moderately elevated. Muscle pathological examinations revealed rimmed vacuoles and intranuclear inclusions (within muscle fibers). The mean duration from onset to diagnosis was (12.33±7.88) years (range: 1 to 32 years). All probands had negative results on conventional next-generation whole-exome sequencing; pathogenic variants were identified through third-generation long-read sequencing or OPDM-targeted repeat-primed polymerase chain reaction(RP-PCR). Among the 43 families, OPDM2 subtype was the most common genetic subtype ( OPDM2 was the predominant subtype in this study. All subtypes share similar age of onset and muscular pathological changes, yet exhibit distinct disease progression patterns. Future multicenter prospective cohort studies are warranted to further elucidate the clinical characteristics, pathogenetic mechanisms, and prognostic differences among OPDM subtypes.
2.Effect of Electroacupuncture on Metabolism-Inflammation Network in Patients with Simple Obesity:A Randomized Controlled Trial
Qi SHU ; Qiumei PENG ; Zhihao XU ; Shanshan FANG ; Wenjing SHAO ; Hongliang CHENG ; Rilong HUANG ; Qing YU ; Ronglin CAI
Journal of Traditional Chinese Medicine 2026;67(13):1415-1421
ObjectiveTo observe the clinical efficacy of electroacupuncture in treatment of simple obesity patients and its effects on the metabolism-inflammation network. MethodsSeventy-two patients with simple obesity were randomly divided into a electroacupuncture group and a sham electroacupuncture group, with 36 patients in each group. In the electroacupuncture group, conventional acupuncture was applied at Zhongwan (CV 12), bilateral Tianshu (ST 25), bilateral Daheng (SP15), Qihai (CV 6), Guanyuan (CV 4), bilateral Zusanli (ST 36) and Fenglong (ST 40). The sham electroacupuncture group used placebo needle at same acupoints as the electroacupuncture group. Both groups were given sparse-dense waves with a frequency of 2/10 Hz, 30 minutes per session, 3 times a week, for a total of 8 weeks of treatment. The changes of human body parameters including body weight, body mass index (BMI), waist circumference, and body fat percentage (BFP), metabolic factors including serum fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL), low density lipoprotein (LDL), leptin, and ghrelin, and inflammatory factors including interleukin-6 (IL-6) and tumor necrosis factor α (TNF-α) were compared before and after treatment. Correlations between changes in leptin, ghrelin and inflammatory factors were further analyzed. ResultsWaist circumference was significantly reduced after treatment in both groups compared with baseline (P<0.05), and the post-treatment waist circumference was lower in the electroacupuncture group than in the sham electroacupuncture group (P<0.05); body weight, BMI and BFP decreased significantly after treatment compared to pretreatment levels in the electroacupuncture group (P<0.05). Serum leptin, IL-6 and TNF-α declined significantly after treatment in both groups versus baseline (P<0.05); the electroacupuncture group had lower post-treatment levels of leptin, IL-6 and TNF-α as well as larger pre-post changes than the sham group (P<0.05); ghrelin increased markedly after treatment in both groups (P<0.05), with higher post-treatment ghrelin and greater pre-post variation observed in the electroacupuncture group (P<0.05). In the electroacupuncture group, changes in ghrelin were negatively correlated with IL-6 and positively correlated with TNF-α (P<0.01); in the sham electroacupuncture group, ghrelin changes were negatively correlated with both IL-6 and TNF-α, whereas leptin changes were positively correlated with the two inflammatory markers (P<0.01). ConclusionElectroacupuncture can effectively improve clinical manifestations, as well as regulate serum metabolic and pro-inflammatory factors in patients with simple obesity, and discrepant correlation patterns of metabolism-inflammation are observed between the two groups.
3.Impact of initial screening strategies on compliance with colonoscopy for colorectal carcinoma in residents aged 50 years and above
Fang XIANG ; Zhihao HU ; Yawei WANG ; Yiying ZHANG ; Fang HUANG ; Qian PENG ; Hongjie YU ; Chaowei FU
Shanghai Journal of Preventive Medicine 2025;37(2):140-144
ObjectiveTo compare colonoscopy compliance rates under different screening strategies, to explore ways to enhance colonoscopy compliance among residents with colorectal carcinoma. MethodsResidents aged between 50‒80 years were recruited through extensive community outreach and voluntary participation. A total of 210 630 residents who participated in the colorectal carcinoma screening program in Jiading District, Shanghai, between 2013 and 2019 were selected as the research subjects. All subjects underwent a colorectal carcinoma risk assessment questionnaire survey and two fecal occult blood tests (FOBT). Positive results in the initial screening were defined as a positive questionnaire survey or a positive result in at least one FOBT. Participants with positive initial screening results were advised to undergo colonoscopy screening in a hospital. Colonoscopy results were collected from hospital reports and physician follow-ups. Compliance with colonoscopy was analyzed under different screening strategies to identify possible factors influencing residents’ willingness to undergo the procedure. ResultsA total of 21 403 individuals (10.16%) were identified as positive with the questionnaire survey, 31 595 individuals (15.00%) tested positive with at least one FOBT. Combined questionnaire and FOBT positivity was observed in 3 501 individuals (1.66%). Among the 48 453 individuals with positive initial screening results, 17 230 (35.56%) underwent colonoscopy, and a total of 315 cases of colorectal cancer were detected. The sensitivity, specificity value of FOBT initial screening were 83.81% and 84.66%, respectively. According to the combined risk assessment and FOBT initial screening preliminary screening, the lowest colonoscopy compliance rate (25.63%) was observed among individuals with only a positive questionnaire, and the highest compliance rate (52.55%) was among those with both positive questionnaire survey and two positive FOBT results. Multivariate analysis revealed that FOBT positivity had the greatest impact on colonoscopy compliance. Those with one positive FOBT test result were 2.64 times more likely to undergo colonoscopy screening than those with negative FOBT results, while individuals with two positive FOBT results were 3.18 times more likely to do so. After adjusting for FOBT results, individuals with positive questionnaire survey results were 1.43 times more likely to undergo colonoscopy screening than those with negative results (95%CI: 1.34‒1.52). Compared to questionnaire-based risk assessment, FOBT results were more influential in determining compliance with colonoscopy. ConclusionThe choice of initial screening method significantly impacts residents’ compliance with colonoscopy. While implementing colorectal carcinoma screening programs, it is necessary to strictly adhere to screening protocols, including risk assessment and FOBT. Additionally, efforts should be made to raise public awareness, encouraging residents to actively participate in risk assessments and FOBT, thereby improving their compliance with colonoscopy.
4.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
5.Isolation and Identification of Staphylococcus xylosus in Nude Mice with Squamous Skin Scurfs
Zhihao KONG ; Xiaofeng WEI ; Lingzhi YU ; Liping FENG ; Qi ZHU ; Guojun SHI ; Chen WANG
Laboratory Animal and Comparative Medicine 2025;45(3):368-375
Objective To isolate pathogenic bacteria from the skin of a nude mouse exhibiting squamous skin scurfs, and perform bacterial identification, traceability analysis, and pathogenicity studies to provide a new approach for the diagnosis of pathogens in nude mice with squamous skin scurfs. MethodsSkin swab samples were collected from a nude mouse exhibiting squamous skin scurfs for nucleic acid testing, bacterial isolation and culture, biochemical identification, 16S rDNA gene amplification and sequencing, and whole genome sequencing to construct a phylogenetic tree. Fifteen BALB/c nude mice were randomized into a saline-treated control group, a high-concentration group treated with 1.8×10⁸ CFU/mL of the isolated bacterial suspension, and a low-concentration group treated with 1.8×10⁷ CFU/mL of the isolated bacterial suspension. Pathogenicity was assessed by animal infection experiments and observation of histopathological changes in skin tissue using HE staining. Results The nucleic acid test for Corynebacterium bovis was negative, excluding infection by this organism. The pathogen isolated on mannitol salt agar and blood agar, combined with Gram staining, suggested a Gram-positive Staphylococcus species. The isolated strain was identified by 16S rDNA sequencing and a fully automated microbial identification system as Staphylococcus xylosus. Phylogenetic tree analysis based on whole genome sequencing showed that the strain was most closely related to an isolate from leafy vegetables in South Korea (GenBank GCA_00207825.1). In the high-concentration group, squamous skin scurfs appeared on the head, neck, and back of nude mice on the 17th day post-infection, while in the low concentration group, similar symptoms appeared on the 20th day post-infection and gradually spread to other areas. The scaling symptoms were transient, lasting for 7 days in the high-concentration group and 3 days in the low-concentration group, after which the skin returned to normal. The infection rate was 33.33% in both the high- and low-concentration groups. No significant pathological changes were observed in the skin tissues of infected mice compared to the control group, indicating marked individual differences in the pathogenicity of the strain in nude mice. Conclusion A strain of Staphylococcus xylosus was isolated from the skin of a nude mouse exhibiting squamous skin scurfs. The strain is an opportunistic pathogen that causes transient squamous skin scurfs without significant histopathological changes, and there are individual differences in the sensitivity of nude mice to this strain. These findings can provide valuable data for pathogen identification in immunodeficient or gene knockout mice.
6.Clinical value of ductoscopy,mammography,and high-frequency color Doppler ultrasonography in diagnosis of nipple bloody discharge combined with intraductal space occupying lesions
Xiancheng AI ; Zhihao YU ; Xiangyu XU
China Journal of Endoscopy 2025;31(2):55-62
Objective To analyze the clinical significance of ductoscopy,mammography,and high-frequency color Doppler ultrasonography in diagnosis of nipple bloody discharge combined with intraductal space occupying lesions.Methods The medical records of 198 patients with nipple bloody discharge combined with intraductal space occupying lesions who underwent surgical treatment from January 2023 to December 2023 were retrospectively analyzed,all the subjects were examined by ductoscopy,mammography and high-frequency color Doppler ultrasonography before operation.The results of postoperative pathology were used as the gold standard to compare and evaluate the efficacy of single and combined use of the above three methods in the diagnosis of nipple bloody discharge combined with intraductal space occupying lesions.Results The results of postoperative pathological diagnosis showed that 152 cases of intraductal benign lesions were intraductal papilloma of the breast.There were 46 malignant lesions,of which 25 were intraductal carcinoma and 21 were invasive ductal carcinoma.The detection rates of nipple bloody discharge combined with intraductal space occupying lesions were 95.45%,81.31%,84.85%and 97.47%,respectively,by ductoscopy,mammography,high-frequency color Doppler ultrasonography and combined examination.Among them,the detection rate of ductoscopy and combined examination were significantly higher than that of mammography and high-frequency color Doppler ultrasonography(P<0.05),there was no significant difference between the detection rate of ductoscopy and combined examination(P>0.05),and there was no significant difference between the detection rate of mammography and high-frequency color Doppler ultrasonography(P>0.05).The sensitivity,specificity,and accuracy of diagnosing nipple bloody discharge combined with intraductal space occupying lesions using ductoscopy were 91.30%,97.20%,and 95.77%,respectively.The above indicators of mammography were 65.00%,94.21%,and 86.96%,respectively.The above indicators of high-frequency color Doppler ultrasonography were 70.73%,96.06%,and 89.88%,respectively.The above indicators of combination of the three methods were 95.65%,97.28%,and 96.89%,respectively.The sensitivity and accuracy of the combined examination of the three methods and the use of ductoscopy alone for the diagnosis of nipple bloody discharge combined with intraductal space occupying lesions were significantly higher than those of mammography and high-frequency color Doppler ultrasonography(P<0.05).However,there was no significant difference between mammography and high-frequency color Doppler ultrasonography(P>0.05),and there was also no significant difference between the combined examination of the three methods and the use of ductoscopy alone(P>0.05).There was no significant difference in diagnostic specificity among the four groups(P>0.05).Conclusion The diagnostic sensitivity and accuracy of ductoscopy for nipple bloody discharge combined with intraductal space occupying lesions are significantly higher than those of mammography and high-frequency color Doppler ultrasonography.The combined examination of the three can improve diagnostic efficiency to a certain extent,but the effect is limited.
7.Clinical value of ductoscopy,mammography,and high-frequency color Doppler ultrasonography in diagnosis of nipple bloody discharge combined with intraductal space occupying lesions
Xiancheng AI ; Zhihao YU ; Xiangyu XU
China Journal of Endoscopy 2025;31(2):55-62
Objective To analyze the clinical significance of ductoscopy,mammography,and high-frequency color Doppler ultrasonography in diagnosis of nipple bloody discharge combined with intraductal space occupying lesions.Methods The medical records of 198 patients with nipple bloody discharge combined with intraductal space occupying lesions who underwent surgical treatment from January 2023 to December 2023 were retrospectively analyzed,all the subjects were examined by ductoscopy,mammography and high-frequency color Doppler ultrasonography before operation.The results of postoperative pathology were used as the gold standard to compare and evaluate the efficacy of single and combined use of the above three methods in the diagnosis of nipple bloody discharge combined with intraductal space occupying lesions.Results The results of postoperative pathological diagnosis showed that 152 cases of intraductal benign lesions were intraductal papilloma of the breast.There were 46 malignant lesions,of which 25 were intraductal carcinoma and 21 were invasive ductal carcinoma.The detection rates of nipple bloody discharge combined with intraductal space occupying lesions were 95.45%,81.31%,84.85%and 97.47%,respectively,by ductoscopy,mammography,high-frequency color Doppler ultrasonography and combined examination.Among them,the detection rate of ductoscopy and combined examination were significantly higher than that of mammography and high-frequency color Doppler ultrasonography(P<0.05),there was no significant difference between the detection rate of ductoscopy and combined examination(P>0.05),and there was no significant difference between the detection rate of mammography and high-frequency color Doppler ultrasonography(P>0.05).The sensitivity,specificity,and accuracy of diagnosing nipple bloody discharge combined with intraductal space occupying lesions using ductoscopy were 91.30%,97.20%,and 95.77%,respectively.The above indicators of mammography were 65.00%,94.21%,and 86.96%,respectively.The above indicators of high-frequency color Doppler ultrasonography were 70.73%,96.06%,and 89.88%,respectively.The above indicators of combination of the three methods were 95.65%,97.28%,and 96.89%,respectively.The sensitivity and accuracy of the combined examination of the three methods and the use of ductoscopy alone for the diagnosis of nipple bloody discharge combined with intraductal space occupying lesions were significantly higher than those of mammography and high-frequency color Doppler ultrasonography(P<0.05).However,there was no significant difference between mammography and high-frequency color Doppler ultrasonography(P>0.05),and there was also no significant difference between the combined examination of the three methods and the use of ductoscopy alone(P>0.05).There was no significant difference in diagnostic specificity among the four groups(P>0.05).Conclusion The diagnostic sensitivity and accuracy of ductoscopy for nipple bloody discharge combined with intraductal space occupying lesions are significantly higher than those of mammography and high-frequency color Doppler ultrasonography.The combined examination of the three can improve diagnostic efficiency to a certain extent,but the effect is limited.
8.A retrospective study on the impact of the number of examined lymph nodes on the survival prognosis of patients with N3b gastric cancer
Xiaodong WANG ; Zhihao YU ; Xintong SUN ; Zhishuo LI ; Xingtu QIN ; Huimin ZHANG ; Yanrui LIANG ; Jing WU ; Mansheng ZHU ; Weihong GUO ; Guoxin LI ; Yanfeng HU ; Liying ZHAO ; Xinhua CHEN
Chinese Journal of Gastrointestinal Surgery 2025;28(10):1141-1150
Objective:To investigate the impact of the number of examined lymph nodes (ELN) on survival outcomes in gastric cancer patients with postoperative pathological stage pN3b.Methods:This retrospective cohort study included 279 pN3b gastric cancer patients who underwent D2 gastrectomy at Nanfang Hospital, Southern Medical University (September 2008 to April 2023), with 35 patients receiving combination chemotherapy and anti-PD-1 therapy (immunotherapy group) and 244 receiving adjuvant chemotherapy alone (nonimmunotherapy group). Additionally, 422 patients with pN3b from the SEER database (2005 to 2020) were collected as an external validation cohort to determine the optimal cutoff value for the number of lymph nodes examined in the nonimmunotherapy group. The primary endpoints were overall survival (OS) and recurrence-free survival (RFS) in the nonimmunotherapy group of the Nanfang Hospital cohort, stratified by whether the number of examined lymph nodes was above or below the ELN optimal cutoff value. These findings were subsequently validated in the SEER cohort.Results:The optimal ELN cutoff value (34 nodes) was determined using X-tile software and by constructing an ELN-HR fitting model with inflection point identification. In the nonimmunotherapy group, patients with ELN >34 exhibited significantly prolonged survival compared to ELN ≤34 (median OS: 25.0 (95%CI:20.5-29.5) to 17.0 (95%CI:12.7-21.3) months, P=0.004; median RFS: 19.0 (95%CI:15.6-22.4) to 13.0 (95%CI:9.5-16.5) months, P=0.048). Multivariate Cox analysis also showed ELN >34 to be an independent protective factor for both OS (HR=0.576, 95%CI: 0.397-0.836) and RFS (HR=0.701, 95%CI: 0.492-0.998). In the SEER cohort, ELN >34 was associated with a 5-month OS extension (19 to 14 months, P=0.065), with multivariate analysis supporting its independent prognostic significance (HR=0.729, 95%CI: 0.580-0.915, P=0.006). Notably, in the immunotherapy group, patients with ELN >34 ( n=30) achieved a median OS of 41 months, but the median OS had not been reached in the ELN ≤34 group ( n=5) (1 death at 48 months). Conclusion:Higher ELN (>34) correlates with improved survival in nonimmunotherapy-treated pN3b gastric cancer patients. However, in pN3b gastric cancer patients treated with immunotherapy, the optimal ELN threshold requires further exploration to determine.
9.Altered patterns of interhemispheric functional connectivity in the cerebral cortex connected via the corpus callosum in Alzheimer's disease
Zheyu LI ; Yifan SHEN ; Renren LI ; Chenxi PAN ; Wei ZHANG ; Zhihao XU ; Qiurong YU ; Dazhi YIN ; Mingxia FAN ; Yunxia LI ; Ruipeng NING
Chinese Journal of Medical Physics 2025;42(11):1434-1444
Objective To investigate alterations in interhemispheric functional connectivity(FC)in the cerebral cortices connected via the corpus callosum in patients with Alzheimer's disease(AD),and to explore their relationships with cognitive function and activities of daily living.Methods Resting-state functional magnetic resonance imaging data were collected from 28 patients with Alzheimer's dementia(d-AD),47 patients with mild cognitive impairment(MCI),and 37 healthy controls(HC).Using a trancallosal tract template,32 pairs of homologous cortical brain regions directly connected to 32 subregions of the corpus callosum were selected as regions of interest for interhemispheric FC analysis.Further correlation analyses were performed between FC values in patient groups and their scores on the Montreal Cognitive Assessment-Basic(MoCA-B)Scale and the Activities of Daily Living(ADL)Scale.Results Compared with HC group,both MCI and d-AD groups exhibited hyperconnectivity(significantly increased FC)in interhemispheric non-homologous brain regions.Specifically,hyperconnectivity in the MCI group was scattered across the frontal,parietal,temporal,and occipital lobes,while in the d-AD group,it was concentrated within the precentral and postcentral gyri.Notably,hyperconnectivity involving the prefrontal and occipital lobes in the MCI group showed significant declines in the d-AD group.The interhemispheric homologous FC in the d-AD group reduced more significantly than the MCI group.Additionally,in the d-AD group,2 interhemispheric FC within the prefrontal lobe(between the bilateral orbital parts of the inferior frontal gyrus,and between the left medial frontal gyrus and the right middle frontal gyrus)were correlated with MoCA-B scores,and 2 FC(between the bilateral middle occipital gyri,and between the left inferior parietal lobule and the right middle frontal gyrus)were correlated with ADL scores.Conclusion MCI and d-AD exhibit distinct patterns of interhemispheric FC alterations,and the interhemispheric FC changes in AD patients are non-progressive.The close relationships between interhemispheric homologous/non-homologous FC and MoCA-B/ADL scores in d-AD patients provide an objective basis and reference for clinical neuromodulation.
10.Early high-sucrose diet exacerbates cognitive impairment in 3×Tg-AD mice via activating cGAS-STING pathway
Xi ZHANG ; Hongyu CHEN ; Jianhao WANG ; Yuke SHI ; Feng GAO ; Yida LYU ; Shuai DING ; Xiang LI ; Hang YU ; Jiabei WANG ; Zhihao WANG
Chinese Journal of Neuromedicine 2025;24(9):873-887
Objective:To investigate the effect of early high-sucrose diet (eHSD) on cognitive function and its regulatory mechanism in 3×Tg-AD mice.Methods:(1) Eighteen specific-pathogen-free (SPF)-grade 2-month-old wide-type (WT) mice were randomly divided into a WT+normal chow diet (NCD) group and a WT+eHSD group, with 9 mice in each group; and 18 SPF-grade 2-month-old 3×Tg-AD mice were randomly divided into a 3×Tg-AD+NCD group and a 3×Tg-AD+eHSD group, with 9 mice in each group. At 2-5 months old, mice in the 4 groups received standard laboratory food+purified water or 30% sucrose water, followed by standard feed for all groups. At 8 months old, cognitive function was assessed by Morris water maze test; fluorescent intensity of AT8 (phosphorylated [p]-tau) and T22 (tau oligomers) in the hippocampal tissues was detected by immunofluorescent staining; concentrations of β-amyloid protein (Aβ) 42 and Aβ 40 were detected by enzyme-linked immunosorbent assay (ELISA); protein expressions of stimulator of interferon genes (STING), TANK-binding kinase 1 (TBK1), p-TBK1, and CCAAT/enhancer-binding protein β (C/EBPβ) were detected by Western blotting; activity of C/EBPβ transcription factor was detected by activity assay; mitochondrial DNA (mtDNA) content in the cytoplasm of cell was detected by real-time quantitative PCR (qPCR). (2) Eighteen SPF-grade 2-month-old 3×Tg-AD mice were randomized into a 3×Tg-AD+eHSD+H-151 group and a 3×Tg-AD+eHSD+dimethyl sulfoxide (DMSO) group, with 9 mice in each group. Mice at 2-5 months old were given standard laboratory food+30% sucrose water; they were, respectively, injected intraperitoneally with STING pathway inhibitor H-151 or DMSO at 5 months old, and continually injected until 8 months old; and then, the behavioral testing, immunofluorescent staining, ELISA, Western blotting and C/EBPβ transcription factor activity experiments were repeated as before. (3) After crossing C/EBPβ heterozygous knockout (C/EBPβ +/-) mice with 3×Tg-AD mice, 3×Tg-AD/C/EBPβ +/- mice were obtained, and 3×Tg-AD mice were used as controls; they were named 3×Tg-AD/C/EBPβ +/-+eHSD group and 3×Tg-AD+eHSD group, with 9 mice in each group. Both groups of mice were given standard laboratory food+30% sucrose water at 2-5 months old, followed by standard feed until 8 months old; and then, the behavioral testing, immunofluorescent staining, ELISA, and Western blotting experiments were repeated as before. (4) C/EBPβ transgenic mice (C/EBPβTg) were crossed with 3×Tg-AD mice to obtain C/EBPβTg/3×Tg-AD mice, and Non-Tg/3×Tg-AD mice were used as controls; they were, respectively, named as C/EBPβTg/3×Tg-AD+eHSD+H-151 group, Non-Tg/3×Tg-AD+eHSD+H-151 group, and Non-Tg/3×Tg-AD+eHSD+DMSO group, with 9 mice in each group. All 3 groups of mice were given standard laboratory food+30% sucrose water at 2-5 months old; at 5-8 months old, mice in the C/EBPβTg/3×Tg-AD+eHSD+H-151 group and Non-Tg/3×Tg-AD+eHSD+H-151 group were intraperitoneally injected with H-151, while mice in the Non-Tg/3×Tg-AD+eHSD+DMSO group were injected with DMSO; and then, the behavioral testing, immunofluorescent staining, ELISA, and Western blotting experiments were repeated as before. Results:(1) Compared with those in the WT+NCD group and WT+eHSD group, area under the latency curve of 3×Tg-AD+eHSD mice was significantly increased, and proportion of time spending in the targeted quadrant of mice in the 3×Tg-AD+NCD group and 3×Tg-AD+eHSD group was significantly decreased ( P<0.05); compared with that in the 3×Tg-AD+NCD group, proportion of time spending in the targeted quadrant in mice of the 3×Tg-AD+eHSD group was significantly reduced ( P<0.05). Compared with the 3×Tg-AD+NCD group, the 3×Tg-AD+eHSD group had significantly increased p-tau and tau oligomers, Aβ 42 and Aβ 40 concentrations in the hippocampus (AT8 fluorescent intensity: 1.000±0.076 vs. 2.902±0.399; T22 fluorescent intensity: 1.000±0.145 vs. 2.495±0.273; Aβ 42: 1.000±0.167 vs.1.956±0.132; Aβ 40: 1.000±0.226 vs.1.900±0.116), significantly increased C/EBPβ protein expression and C/EBPβ transcription factor activity (1.000±0.164 vs. 1.804±0.112; 1.000±0.216 vs. 2.743±0.301), and statistically increased mtDNA level detected by D-loop1 and D-loop3 (1.000±0.234 vs. 2.800±0.210; 1.000±0.155 vs. 2.952±0.078; P<0.05). Compared with the 3×Tg-AD+NCD group, the 3×Tg-AD+eHSD group had significantly increased STING protein expression and p-TBK1/TBK1 ratio (STING: 1.000±0.192 vs. 2.093±0.081; p-TBK1/TBK1: 1.000±0.148 vs. 1.561±0.112, P<0.05). (2) Compared with the 3×Tg-AD+eHSD+DMSO group, the 3×Tg-AD+eHSD+H-151 group had significantly decreased area under the latency curve, significantly increased proportion of time spending in the targeted quadrant, significantly decreased p-tau and tau oligomers expressions, Aβ 42 and Aβ 40 concentrations in the hippocampus (AT8 fluorescent intensity: 1.000±0.142 vs. 0.538±0.057; T22 fluorescent intensity: 1.000±0.104 vs. 0.665±0.088; Aβ 42: 1.000±0.084 vs. 0.600±0.007; Aβ 40: 1.000±0.138 vs. 0.476±0.083), significantly decreased STING protein expression and p-TBK1/TBK1 ratio (STING: 1.000±0.054 vs. 0.468±0.111; p-TBK1/TBK1: 1.000±0.057 vs. 0.598±0.090), and significantly decreased C/EBPβ transcription factor activity (1.000±0.097 vs. 0.445±0.106; P<0.05). (3) Compared with the 3×Tg-AD+eHSD group, the 3×Tg-AD/C/EBPβ +/-+eHSD group had significantly decreased area under the latency curve, significantly increased proportion of time spending in the targeted quadrant, significantly decreased p-tau and tau oligomers, Aβ 42 and Aβ 40 concentrations in the hippocampus (AT8 fluorescent intensity: 1.000±0.160 vs. 0.506±0.065; T22 fluorescent intensity: 1.000±0.127 vs. 0.346±0.048; Aβ 42: 1.000±0.017 vs. 0.510±0.101; Aβ 40: 1.000±0.098 vs. 0.586±0.153), and significantly decreased C/EBPβ protein expression (1.000±0.101 vs. 0.568±0.094; P<0.05). (4) Compared with the Non-Tg/3×Tg-AD+eHSD+DMSO group, the Non-Tg/3×Tg-AD+eHSD+H-151 group had significantly decreased area under the latency curve, significantly increased proportion of time spending in the targeted quadrant, and significantly decreased p-tau and tau oligomers expressions, Aβ 40 concentration in the hippocampus, and the Non-Tg/3×Tg-AD+eHSD+H-151 group, the C/EBPβTg/3×Tg-AD+eHSD+H-151 group had significantly decreased STING protein expression and p-TBK1/TBK1 ratio in the hippocampus ( P<0.05). Compared with the Non-Tg/3×Tg-AD+eHSD+H-151 group, the C/EBPβTg/3×Tg-AD+eHSD+H-151 group had significantly increased area under the latency curve, significantly decreased proportion of time spending in the targeted quadrant, and significantly increased p-tau and tau oligomers expressions, Aβ 40 and Aβ 42 concentration in the hippocampus ( P<0.05). Conclusion:The eHSD aggravates cognitive impairment in 3×Tg-AD mice through activating cGAS-STING-C/EBPβ pathway.

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