1.Impact of metabolic risk factor control on mortality risk in patients with metabolic dysfunction-associated steatotic liver disease
Jingdan ZHANG ; Bingbing WANG ; Zeran WANG ; Fan FENG ; Ren NA ; Hongyan GE
Journal of Clinical Hepatology 2026;42(7):1576-1585
ObjectiveTo investigate the association of comprehensive control of multiple metabolic risk factors with the risk of all-cause mortality in patients with metabolic dysfunction-associated steatotic liver disease (MASLD) and its quantitative effect. MethodsA retrospective cohort study was conducted based on the data from National Health and Nutrition Examination Survey (NHANES) in 2003—2018. A total of 29 458 participants were enrolled, including 12 528 patients with MASLD and 16 930 non-MASLD controls. The control status of nine risk factors was assessed, i.e., hypertension, diabetes, liver fibrosis (assessed by fibrosis-4 index [FIB-4]), aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio, serum albumin, renal function (assessed by serum creatinine), systemic inflammation (assessed by neutrophil count), smoking, and physical activity. According to the status of comprehensive risk factor control, the patients with MASLD were further divided into low-control group with 1 107 patients, moderate-control group with 7 042 patients, and high-control group with 4 379 patients. A total of 397 participants were enrolled in the external validation cohort, among whom there were 266 participants with MASLD and 131 participants without MASLD. The Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and the chi-square test was used for comparison of categorical data between groups. The Cox proportional hazards model incorporating complex sampling weights for NHANES data were used to investigate the association between the degree of risk factor control and all-cause mortality, and the Kaplan-Meier curve, the log-rank test, restricted mean survival time (RMST), and population attributable fraction (PAF) were used for further assessment. The robustness of the findings was validated in an independent clinical cohort (n=397). ResultsThe cohort study based on the NHANES database showed that during the median follow-up time of 6.7 years, 1 171 deaths occurred among the 12 528 patients with MASLD. There was a significant reduction in mortality risk with the increase in the number of risk factors controlled (trend test: χ2=65.06, P<0.001). Compared with the low-control group, the mortality risk decreased successively in the moderate-control group (hazard ratio [HR]=0.59, 95% confidence interval [CI]: 0.51 — 0.69) and the high-control group (HR=0.36, 95%CI: 0.27 — 0.48), and the mortality risk was reduced by 22% for each additional risk factor controlled (HR=0.78, 95%CI: 0.74 — 0.82). The PAF analysis showed that AST/ALT ratio control (PAF=19.2%) and physical activity (PAF=18.9%) were the largest contributors. The survival analysis of the sub-cohort constructed based on nearest neighbor matching (with a maximum follow-up time of 13.3 years) showed that there was a significant difference in cumulative survival rate between the MASLD groups with different control levels and the non-MASLD group (P<0.001). The RMST analysis further quantified the survival benefit of each group, and the high-control group had the longest 10-year RMST (9.88 years), which was superior to that of the non-MASLD group (9.43 years), the moderate-control group (9.33 years), and the low-control group (7.96 years). The multivariable-adjusted model analysis of the complete cohort showed that compared with the non-MASLD population, the MASLD patients in the low-control group had a significant increase in mortality risk, the moderate-control group had a comparable mortality risk, and the high-control group had a significantly lower mortality risk. The subgroup analysis showed a significant protective effect of comprehensive control in both male and female individuals (P<0.05), and the interaction analysis suggested a potentially stronger protective effect in female individuals (Pfor interaction=0.031). The protective effect was statistically significant in patients aged ≥60 years (P<0.05). There was a significant difference in protective effect across the groups stratified based on waist circumference (P<0.05), but there was no significant interaction between groups (Pfor interaction=0.821). In the external validation cohort, among the 397 participants, there were 72 cases of all-cause mortality (18.1%). The survival analysis showed a significant gradient increase in the survival rate of MASLD patients with the increase in the level of control (P=0.022). The RMST analysis showed that the moderate- and high-control groups had a better RMST than the non-MASLD group at 5 years (4.47 years vs 4.56 years vs 4.40 years), with a more significant survival advantage at 10 years (8.67 years vs 8.95 years vs 8.44 years). The Cox regression analysis showed that, with the low-control MASLD group as the reference, there was a significant reduction in mortality risk in the moderate-control group (HR=0.41, 95%CI: 0.21 — 0.81) and the high-control group (HR=0.32, 95%CI: 0.14 — 0.71); with the non-MASLD population as the reference, there was a significant increase in mortality risk in the low-control group (HR=2.02, 95%CI: 1.03 — 3.95), with a comparable mortality risk in the moderate-control group (HR=0.83, 95%CI: 0.48 — 1.44) and the high-control group (HR=0.66, 95%CI: 0.30 — 1.38), and the point estimates of the effects were highly consistent with the main study. ConclusionAmong MASLD patients, better control of metabolic risk factors is associated with a lower mortality risk. Achieving optimal control can eliminate the excess mortality risk associated with MASLD, thereby helping patients achieve a better survival prognosis than the general population.
2.Cancer-associated fibroblasts mediate migration of myeloid-derived suppressor cells in pancreatic ductal adenocarcinoma through SDF-1/CXCR4 pathway
Bingbing ZHANG ; Hao HU ; Yuchuan SHI ; Xuefei LIU ; Zhi ZHU ; Jing ZHANG
Academic Journal of Naval Medical University 2025;46(7):838-846
Objective To explore the mechanism by which cancer-associated fibroblasts(CAFs)regulate CD13-high expression neutrophil-like myeloid-derived suppressor cell(CD13hi-nMDSC)migration in pancreatic ductal adenocarcinoma(PDAC),so as to provide potential molecular targets and experimental evidences for immunotherapy in patients.Methods CAFs were isolated and purified from pancreatic cancer tissues of 5 PDAC patients.The phenotype and purity of CAFs were identified by immunofluorescence and flow cytometry.The expression of related factors in CAF was detected by quantitative polymerase chain reaction(qPCR)and enzyme-linked immunosorbent assay(ELISA).CAF conditioned medium and myeloid-derived suppressor cell(MDSC)migration system were constructed by Transwell to observe the migration of MDSCs and to study the specific mechanisms by which the aforementioned cytokines participate in regulating the migration of MDSCs.Results The isolated primary CAFs expressed activation biomarkers fibroblast activation protein(FAP)and α-smooth muscle actin(α-SMA),while the human foreskin fibroblasts(HFFs)of control cells did not express FAP and α-SMA.qPCR results showed that the mRNA expression levels of interleukin 6(IL-6),monocyte chemotactic protein 1(MCP-1),and stromal cell-derived factor 1(SDF-1)in CAFs were higher than those in HFFs(all P<0.01).The contents of IL-6,MCP-1,and SDF-1 in the CAF culture supernatant were significantly higher than those in the HFF culture supernatant(all P<0.01),and the secretion content increased with the prolongation of culture time.Compared with HFF conditioned medium and regular medium(RPMI 1640),CAF conditioned medium could recruit more total MDSCs and CD13hi-nMDSCs(all P<0.01).The addition of SDF-1 recombinant protein alone in the culture system could induce the migration of total MDSCs and CD13hi-nMDSCs,and the addition of SDF-1 neutralizing antibodies or C-X-C motif chemokine receptor 4(CXCR4)blocking antibodies could significantly reduce the migration of CD13hi-nMDSCs induced by CAF conditioned medium(all P<0.01).Although MCP-1 alone could also induce the migration of total MDSCs and CD13hi-nMDSCs,the number of CD13hi-nMDSCs migrating was significantly less than that of the SDF-1 experimental group.The IL-6 recombinant protein did not induce the migration of total MDSCs or CD13hi-nMDSCs.Conclusion CAFs can mediate the migration of total MDSCs and CD13hi-nMDSCs in PDAC through SDF-1/CXCR4 pathway.
3.Effects of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver
Yaqi CHANG ; Tianjiao YANG ; Yunfei LUAN ; Jihang LI ; Zexin ZHANG ; Bingbing ZHANG ; Chuang XU ; Wei YANG
Chinese Journal of Veterinary Science 2025;45(10):2282-2291
To explore the impacts of TGR5 on liver lipid metabolism and bile acid synthesis in dairy cows with fatty liver.Liver tissues of healthy cows and cows with fatty liver were collected through puncture technique.The protein and mRNA expressions of lipid synthesis-related factors ACC1,FAS,SREBF1,lipid oxidation factor CPT1A,and bile acid synthesis-related factors CYP8B1,CYP7B1,CYP27A1 were detected by Western blot and fluorescent quantitative PCR.Moreover,the mRNA levels of CYP7B1 were determined.Primary hepatocytes of 1-day-old calves were extracted and cultured in vitro,and four treatment groups were established,namely Control,NEFA,INT-777,and the INT-777+NEFA group.The concentration of NEFA group was 1.2 mmol/L,the con-centration of INT-777 group was 1 μmol/L,and the concentration of INT-777+NEFA group was 1.2 mmol/L NEFA and 1 μmol/L INT-777 simultaneously.After 12 h of stimulation,cells were collected,and the protein and mRNA levels of ACC1,FAS,SREBF1,CPT1A,CYP8B1,CYP7A1,CYP27A1,and the mRNA levels of CYP7B1 were detected by Western blot and fluorescent quanti-tative PCR.The content of lipid droplets and TG in the cells were detected by flow cytometry and kit.The results demonstrated that compared with healthy cows,the protein and mRNA expressions of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the liver tissues of fatty liver cows were upreg-ulated,while the protein and mRNA levels of CPT1 A,CYP27A1,TGR5,and the mRNA levels of CYP7B1 were downregulated.In vitro experiments revealed that compared with the Control group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,and CYP7A1 in the NEFA group were upregulated,and the protein and mRNA levels of CPT1A,CYP27A1,and TGR5,as well as the mRNA level of CYP7B1,were downregulated.Compared with the NEFA group,the protein and mRNA levels of ACC1,FAS,SREBF1,CYP8B1,CYP7A1 were downregulated in the INT-777+NEFA group,while the protein and mRNA levels of CPT1A CYP27A1,and TGR5 as well as the mRNA level of CYP7B1,were upregulated.The results of flow cytometry and the kit indicated that the lipid droplets and TG content in the NEFA group were upregulated compared with the Control group,while the lipid droplets and TG content in the INT-777+NEFA group were downregulated compared with the NEFA group.The above results suggested that the addition of TGR5 agonist promoted the expression of TGR5 and ameliorated the abnormal lipid metabolism and bile acid synthesis in the liver of dairy cows with fatty liver.
4.Epidemiological Characteristics of Malignant Tumors in Cancer Registration Areas of Heilongjiang Province in 2019 and the Trend from 2013 to 2019
Wanying WANG ; Huixin SUN ; Maoxiang ZHANG ; Haihan JIA ; Min ZHAO ; Guohong GAO ; Bingbing SONG
China Cancer 2025;34(5):368-376
[Purpose]To analyze the incidence and mortality of malignant tumors in cancer regis-tration areas of Heilongjiang Province in 2019 and the trend from 2013 to 2019.[Methods]The incidence and mortality data of malignant tumors reported by the Heilongjiang provincial cancer registries from 2013 to 2019 were collected,and the quality of data was assessed.The crude in-cidence/mortality rate,age-standardized incidence/mortality rate by Chinese standard population(ASIRC/ASMRC)and world standard population(ASIRW/ASMRW),0~74 years old cumulative rate were calculated.Joinpoint 4.6.0 software was used to calculate the average annual percentage change(AAPC)of ASIRC/ASMRC for the trend analysis from 2013 to 2019.[Results]In 2019,there were 16 732 new cases of malignant tumors in the cancer registration areas of Heilongjiang Province,including 8 639 males and 8 093 females.The crude incidence rate was 295.37/105,with an ASIRC and ASIRW of 167.10/105 and 164.18/105,respectively.There were 10 988 malig-nant tumor deaths,including 6 540 males and 4 448 females.The crude mortality rate was 193.97/105,with an ASMRC and ASMRW of 101.22/105 and 101.66/105,respectively.The inci-dence and mortality of malignant tumors increased rapidly after the age of 55,and the incidence and mortality of males were slightly higher than those of females.The top five malignant tumors of high incidence were lung cancer,female breast cancer,colorectal cancer,liver cancer and thy-roid cancer,and the top five malignant tumors of high mortality were lung cancer,liver cancer,colorectal cancer,stomach cancer and female breast cancer.From 2013 to 2019,the ASIRC of malignant tumors in cancer registration areas increased from 153.08/105 in 2013 to 167.10/105 in 2019,and the ASMRC increased from 92.22/105 in 2013 to 101.22/105 in 2019,but there was no statistical difference in the change trend.[Conclusion]The incidence and mortality of malignant tumors in Heilongjiang Province remain high.Lung cancer,female breast cancer,colorectal can-cer,liver cancer and stomach cancer should be the focus of cancer prevention and control.
5.Stearic acid affects the expression of IL-17 in CD4+T cells from ketosis cows through CD36
Ziwei JI ; Siyao LI ; Haixin ZHANG ; Ziwei LI ; Shangmingzhu ZHANG ; Wei YANG ; Chuang XU ; Bingbing ZHANG
Chinese Journal of Veterinary Science 2025;45(3):602-610
The peripheral blood of healthy or ketosis dairy cows was collected,and CD4+T cells were isolated.The expressions of lipid synthesis related proteins fatty acid synthase(FASN),acetyl coenzyme A carboxylase 1(ACC1),cluster of differentiation 36(CD36)and store-operated calcium entry(SOCE)related proteins ORAIl,ORAI2,ORAI3,STIM1,STIM2 were detected by Western blot.IL-17 cells were detected by flow cytometry.CD4+T cells were isolated from the spleen of 1-day-old calves and cultured in vitro.Cells were treated and divided into control(Ctrl)group,si-lenced CD36(siCD36)group,stearic acid(SA)group,and SA+siCD36 group.Cells in the Ctrl and SA groups were transfected with 75 pmol/L negative control siRNA for 48 h,and then stimulated with 200 μmol/L SA for 24 h;Cells in the siCD36 group and SA+siCD36 group were transfected with 75 pmol/L CD36 siRNA for 48 h,and then stimulated with 200 μmol/L SA for 24 h in the SA+siCD36 group.The protein expression of FASN,CD36,ACC1,ORAI1,ORAI2,ORAI3,STIM1 and STIM2 was detected by Western blot,and IL-17 cells were detected by flow cytometry.The results showed that the expression of IL-17 in peripheral blood CD4+T cells of ketosis dairy cows was significantly increased compared to that of healthy cows(P<0.01).Additionally,the protein level of FASN,CD36,STIM1(P<0.05),and ACC1,ORAI2,ORAI3,STIM2(P<0.01)were up-regulated.Compared with the Ctrl group,the protein expression levels of CD36,ACC1 and ORAI3(P<0.05)were up-regulated in the SA group,as well as the protein expression of FASN and STIM1(P<0.01).Additionally,the expression of IL-17 was significantly increased(P<0.05).Compared with the SA group,there was a decrease in the protein expression of STIM1,ORAI1(P<0.05)and CD36,ACC1,FASN,ORAI2(P<0.01)in the siCD36+SA group,as well as IL-17(P<0.05).These results suggest that SA can promote the expression of IL-17 in CD4+T cells in ketosis cows by regulating fatty acid synthesis and activating SOCE channels through CD36.
6.Vitamin D3 on neutrophil calcium uptake and LC3 autophagy function in hypocal-cemic dairy cows
Xinquan LYU ; Siyao LI ; Bingbing ZHANG ; Chuang XU ; Wei YANG
Chinese Journal of Veterinary Science 2025;45(6):1268-1272
To investigate the effect of VD3 on the concentration of Ca2+in neutrophils of subclinical hypocalcemia dairy cows and the autophagy of LC3,peripheral blood of dairy cows was collected and neutrophils were isolated and extracted.Western blot was used to detect the abundance of LC3 autophagy pathway-related proteins(LC3-Ⅱ,P62,ATG5)in cells.The intracellular Ca2+concen-tration of neutrophils was detected by flow cytometry.The LC3 autophagosome was detected by la-ser confocal technology.By adding VD3 to affect intracellular Ca2+concentration levels,the effects of vitamin D3 on calcium absorption and LC3 autophagy function were investigated.Western blot result showed that the protein abundances of LC3 and ATG5 in neutrophils of subclinical hypocal-cemia dairy cows decreased,while the protein abundance of p62 increased,indicating that the auto-phagy function of LC3 in subclinical hypocalcemia dairy cows was weakened.After VD3 treatment,the flow cytometry results showed that VD3 promoted the increase of Ca2+concentration levels in neutrophils.The Western blot results showed that the LC3 autophagy function of neutrophils from healthy cows and subclinical hypocalcemia cows was enhanced under the action of VD3,indicating that VD3 promoted the concentration of Ca2+in neutrophils and then affected the LC3 autophagy function.
7.Efficiency of Rituximab for the treatment of the initial episode of steroid-sensitive nephrotic syndrome in children
Tingting YUAN ; Bingbing ZHU ; Yan LI ; Qianqian PENG ; Huandan YANG ; Na CHEN ; Zhaowen ZHONG ; Ruifeng ZHANG
Chinese Journal of Applied Clinical Pediatrics 2025;40(2):125-129
Objective:To evaluate the efficacy and safety of Rituximab (RTX) combined with short-term use of glucocorticoids in the treatment of the initial episode of steroid-sensitive nephrotic syndrome (SSNS).Methods:A retrospective case summary.A total of 30 children with SSNS treated in the Department of Intrarenal Rheumatism and Immunology, Xuzhou Children′s Hospital from December 2021 to March 2023, were enrolled in this study.They were given a standard dose of RTX (375 mg/m 2) and glucocorticoids for a short term.The patients were followed up for 1 year, and the general condition, changes in CD19 + B lymphocytes expression after RTX treatment, the relapse rate, the median relapse-free survival and adverse reactions of RTX were recorded.Kaplan-Meier method was used to analyze the relapse-free survival time. Results:CD19 + B lymphocytes were depleted 2 weeks after RTX treatment, and the median time required for CD19 + B lymphocytes reconstitution was 5 months after RTX treatment.The 1-year relapse-free rate was 90.00%, and the median relapse-free survival was 11 months.The glucocorticoid discontinuation rate was 93.33% in 3 months after RTX treatment, and 100% in 6 and 12 months after RTX treatment.Adverse reactions included infusion reactions in 7 cases (23.33%), neutropenia/leukopenia in 5 cases (16.67%), hypoimmunoglobulinemia in 10 cases (33.33%), infections in 4 cases.One case was complicated with severe influenza A virus infection after CD19 + B lymphocytes reconstruction. Conclusions:A standard dose of RTX can significantly reduce the relapse frequency, maintain long-term remission of proteinuria, and reduce the dose of glucocorticoids in patients at the initial episode of SSNS.Thus, it is worthy of clinical promotion.
8.Effect of miRNA-381-3p/MuRF1 axis on cardiopulmonary injury in mice with hypoxic pulmonary hypertension
Bin WU ; Zigeng YANG ; Ling JIN ; Jing ZHANG ; Hongmei WEI ; Bingbing CAI ; Yuying WEI
Tianjin Medical Journal 2025;53(6):571-577
Objective To explore the effect of microRNA-381-3p(miR-381-3p)/MuRF1 axis on cardiopulmonary injury in hypoxia-induced pulmonary hypertension(HPH)mice and its potential mechanisms.Methods Sixty mice were randomly assigned to four groups:the normal control group(NC),the hypobaric hypoxia-induced pulmonary hypertension(HPH)group,the HPH+agomir control group and the HPH+miR-381-3p agomir analog group(HPH+miR-381-3p agomir),with 15 mice in each group.The HPH mouse model was established using a low-pressure and hypoxic artificial chamber.Three weeks prior to the establishment of the HPH model,miR-381-3p agomir and its corresponding control agomir were prepared by dissolving them in RNA-free phosphate-buffered saline(PBS)according to the experimental requirements.These solutions were administered via tail vein injection at a dose of 10 mg/kg,twice weekly for three consecutive weeks.Right heart function was assessed using echocardiography.Right ventricular systolic pressure(RVSP)was measured via cardiac catheterization.Pulmonary vascular remodeling was evaluated through hematoxylin and eosin(HE)staining.Levels of inflammatory cytokines in bronchoalveolar lavage fluid were quantified using enzyme-linked immunosorbent assay(ELISA).Real-time quantitative fluorescent PCR(RT-qPCR)was employed to analyze the mRNA expression levels of miR-381-3p and MuRF1.Potential targets of miR-381-3p were predicted,and pathway enrichment analysis was conducted.A dual-luciferase reporter gene assay was performed to confirm the direct regulatory effect of miR-381-3p on MuRF1.Results Compared with the NC group,the mRNA expression of miR-381-3p was significantly decreased in both the HPH group and the HPH+agomir control group,whereas the mRNA expression of MuRF1 was significantly increased(P<0.05).In contrast,compared with the HPH group and the HPH+agomir control group,the mRNA expression of miR-381-3p was significantly increased in the HPH+miR-381-3p agomir group,while the mRNA expression of MuRF1 was significantly decreased(P<0.05).Additionally,compared with the NC group,RVSP,right ventricular anterior wall thickness(RVAW),right ventricular hypertrophy index(RVHI),right ventricular collagen volume fraction(CVF),distal pulmonary artery wall thickness ratio(WT),pulmonary artery wall area ratio(WA),as well as IL-1β,IL-6 and TNF-α levels in alveolar lavage fluid were significantly increased in the HPH group and the HPH+agomir control group,whereas the right ventricular diameter(RVID)was significantly decreased(P<0.05).Conversely,compared with the HPH group and the HPH+agomir control group,RVSP,RVAW,RVHI,right ventricular CVF,WT,Wa and RVID were decreased in the HPH+miR-381-3p agomir group,and IL-1β,IL-6,and TNF-α levels of alveolar lavage fluid were significantly decreased(P<0.05).Furthermore,the downstream target genes of miR-381-3p were predicted in the database,and MuRF1 was a potential target,and the Cytoskeleton in muscle cells ranked first in the significant enrichment of target genes.Compared with WT-MuRF1+mimic control group,the luciferase activity was decreased in the WT-MuRF1+miR-381-3p mimic group(P<0.05).There was no significant difference in the luciferase activity between the Mut-MuRF1+mimic control group and the Mut-MuRF1+miR-381-3p mimic group.Conclusion Overexpression of miR-381-3p can improve cardiopulmonary injury in HPH mice,and the mechanism may be related to the targeted inhibition of MuRF1 by miR-381-3p.
9.Chiral analysis of dextromethorphan and levomethorphan in human hair and urine
Qiao YANG ; Jian ZHANG ; Rui HU ; Bingbing TIAN ; Zhonghui ZHANG
Chinese Journal of Forensic Medicine 2025;40(3):267-272
Objective To establish a method for chiral analysis of dextromethorphan and levomethorphan in hair and urine by high performance liquid chromatography-triple quadrupole mass spectrometry.Methods Hair samples were extracted by grinding ultrasound,and urine samples were extracted by acetonitrile.The supernatant were separated using a Astec CHIROBIOTIC? V2 column(25 cm × 4.6 mm,5 μm)with 0.5 ‰ formic acid and 10 mmol/L ammonium acetate-methanol as mobile phases,then analyzed in scheduled-multiple reaction monitoring mode.Results The limits of detection and lower limits of quantitation of dextromethorphan and levomethorphan were 0.001 ng/mg and 0.005 ng/mg in hair samples,while they were 0.050 ng/mL and 0.200 ng/mL in urine samples.In the corresponding concentration range,there were good linearity for chiral isomers with correlation coefficients over 0.999.Besides,the recovery rates,accuracy and precision met analytical requirements,and no significant matrix effect were exhibited.In the actual case samples,dextromethorphan were detected in hair of 12 cases and in the corresponding urine of 3 cases.Conclusion This method is rapid,sensitive and reliable for chiral analysis of dextromethorphan and levomethorphan in hair and urine.
10.Correlation between cerebral perfusion and cognitive function in patients with minor stroke or transient ischemic attack caused by severe intracranial arterial stenosis or occlusion
Meiling SHANG ; Yanran CHEN ; Bingbing GUO ; Xiaotong CHI ; Lu QUAN ; Gezhi YAN ; Hui WANG ; Ling MA ; Fude LIU ; Jia YU ; Jianfeng HAN ; Ming ZHANG ; Wanghuan DUN ; Yujing WANG
Chinese Journal of Cerebrovascular Diseases 2025;22(10):701-711
Objective This study aimed to investigate the correlation of cerebral perfusion and cognitive function status in patients with minor stroke(MS)or transient ischemic attack(TIA)complicated by severe intracranial arterial stenosis or occlusion(hereafter referred to as ICAS-MSTIA).Methods Retrospectively enrol consecutive ICAS-MSTIA patients admitted to the Department of Neurology,the First Affiliated Hospital of Xi'an Jiaotong University,from June 2023 to May 2024.In the meantime,healthy controls were openly recruited.The ICAS-MSTIA patients were divided into two groups based on the side of intracranial large artery stenosis or occlusion:the left intracranial large artery involvement group and the right intracranial large artery involvement group.All patients with intracranial large artery stenosis or occlusion underwent MR scanning within 2 weeks after the first episode of TIA or MS,while there was no specific time requirement for MR examination in the healthy control group.On the day of MR scanning,the Montreal cognitive assessment(MoCA)scale was used to evaluate the participants'global cognitive function and performance in various cognitive domains,including visuospatial/executive function,naming,attention,language,abstraction,delayed recall,and orientation.General information of all participants was collected,including age,sex,educational level,body mass index,and history of smoking and alcohol consumption.Clinical data were collected from both left and right intracranial large artery involvement groups,including cerebrovascular risk factors(such as,diabetes mellitus,hypertension,and hyperlipidemia),National Institutes of Health stroke scale(NIHSS)score at admission,responsible stenotic or occluded arteries(internal carotid artery,middle cerebral artery),degree of stenosis in the responsible vessel(severe stenosis[stenosis rate 70%-99%],occlusion[stenosis rate100%])and non-responsible vessel(no stenosis[0],mild stenosis[stenosis rate>0-49%]),collateral circulation compensation(American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology[ASTIN/SIR]collateral circulation classification),and responsible events(TIA,MS).General data and MoCA scale scores were compared across the three groups,while clinical data were compared between the left and right intracranial large artery involvement groups.Statistical parametric mapping 12(SPM 12)was used to perform voxel-wise independent samples t-tests on cerebral blood flow(CBF)differences among the left ICAS-MSTIA group,right ICAS-MSTIA group,and healthy control group,with cluster-level family-wise error(FWE)correction applied for adjustment.Multiple linear regression analysis was conducted to evaluate the relationship between global CBF values and total MoCA scores in ICAS-MSTIA patients with left or right intracranial large artery involvement.Results A total of 33 ICAS-MSTIA patients and 33 healthy controls were enrolled in the study.Among the ICAS-MSTIA patients,21 had left intracranial large artery involvement and 12 had right involvement.(1)Among the three groups,statistically significant differences were observed in the proportions of individuals with reported smoking history(P=0.024)and alcohol consumption history(P=0.011).The left intracranial large artery involvement group had a higher NIHSS score(0[0,2]vs.0[0,0],P=0.044)and a higher proportion of patients with internal carotid artery involvement(13/21 cases vs.2/12 cases,P=0.027)compared with the right side group.No statistically significant differences were observed in other general or clinical data across the three groups or between the two non-control groups(all P>0.05).(2)Statistically significant differences were found across the three groups in the MoCA scale total score and scores of visuospatial/executive function,attention,language,abstraction,delayed recall,and orientation cognitive domains(all P<0.05),while no significant difference was noted in the naming score(P=0.063).The left intracranial large artery involvement group had lower total MoCA score and lower scores in visuospatial/executive function,attention,language,abstraction,delayed recall,and orientation in comparison to the healthy control group(all P<0.016 7).The right intracranial large artery involvement group had significantly lower scores in language,abstraction,and orientation domains than the healthy control group(all P<0.016 7).Additionally,the left side group had a lower attention domain score than the right side group(P<0.016 7).No other statistically significant differences were found in pairwise comparisons(all P>0.016 7).(3)Patients in both the left and right intracranial large artery involvement groups exhibited a significant decrease in CBF in extensive regions on the affected side,including the temporal lobe,dorsolateral prefrontal cortex,and occipital lobe.Furthermore,after correction,in the left involvement group CBF was higher in the contralateral lingual gyrus,cuneus,and calcarine sulcus compared with the healthy control group(P<0.05).While in the right involvement group,no regions had increased CBF compared to the healthy control group.(4)Multiple linear regression showed positive correlation between CBF in ipsilateral precentral gyrus and superior temporal gyrus,and the total MoCA score in patients with left intracranial large artery involvement(FWE-corrected,P<0.05).In contrast,there was no correlation between CBF and total MoCA score in patients with right intracranial large artery involvement.Conclusions ICAS-MSTIA patients exhibited various degrees of impairment in cerebral perfusion and cognitive function.A significant positive correlation is observed between these two impairments in patients with left intracranial large artery involvement.

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