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Objective:To investigate the effect of aucubin on behaviors and excessive activation of astrocytic in attention deficit/hyperactivity disorder (ADHD) model mice.Methods:Twelve wild-type C57BL/6 pregnant mice (female, clean grade) were intraperitoneally administered with esketamine (15 mg/kg) to establish an ADHD model in offspring mice. The offspring mice were divided into control+ saline group, control+ aucubin group, Ketamine+ saline group and Ketamine+ aucubin group according to the nest matching principle with 15 in each group.At 14 days after birth, mice in the control+ aucubin group and Ketamine+ aucubin group were administered with aucubin (5 mg/kg, once a day) by gavage for 5 days. Mice in control+ saline group and Ketamine+ saline group were administered with equal volume of 0.9% sodium chloride solution. The offspring mice were housed with their mothers in the same cage until 21 days after birth. Twenty-one days after birth, the offspring mice were evaluated by open field test and elevated plus maze tests. Immunofluorescence assay was used to detect the expression of glutamate decarboxylase 2 (GAD2), γ- aminobutyric acid (GABA) and glial fibrillary acidic protein (GFAP) in the amygdala. The morphological changes of astrocytes were quantitatively analyzed by Sholl analysis. GraphPad Prim 9.0.1 software was used for statistical analysis. The comparison of multiple groups was conducted by one-way ANOVA or Kruskal-Wallis test.Results:(1)The results of behavioral experiments showed that the total distance traveled in the open field test and the residence time in open arm of the elevated plus maze were statistically significant ( F=236.90, H=39.92, both P<0.001). The total distance ((7 044±249)mm, (22 891±2 175)mm, P<0.05) and the residence time in open arm(12.69(9.86, 17.24)s, 2.72(0.57, 3.87)s, P<0.05) of mice in Ketamine+ saline group were both higher than those in control+ saline group.The total distance((22 891±2 175)mm, (8 252±839)mm, P<0.05) and the the residence time in open arm(5.45(1.13, 10.99)s, 12.69(9.86, 17.24)s, P<0.05) of Ketamine+ aucubin group were both lower than those of Ketamine+ saline group.(2)The immunofluorescence results showed that the levels of GAD2, GABA and GFAP intensity in amygdala of mice in the four groups were statistically significant ( F=145.50, 50.08, 53.83, all P<0.05). Compared with control+ saline group, the fluorescence intensities of GAD2 ((100.00±9.60)%, (24.86±4.14)%, P<0.05) and GABA ((100.00±16.84))%, (25.48±5.70)%, P<0.05) of Ketamine+ saline group were down-regulated, and the GFAP((100.00±18.02)%, (223.80±25.85)%, P<0.05) was up-regulated. Compared with Ketamine+ saline group, the fluorescence intensities of GAD2 ((24.86±4.14)%, (56.08±6.55)%, P<0.05) and GABA((25.48±5.70)%, (52.59±15.74)%, P<0.05) in Ketamine+ aucubin group were up-regulated, but the fluorescence intensity of GFAP ((223.80±25.85)%, (157.10±22.10)%, P<0.05) was down-regulated.(3)Sholl analysis indicated that the number of the intersections between the astrocyte processes or the branches of astrocyte processes was statistically significant in the 4 groups ( F=12.47, P<0.05). Compared with control+ saline group, the number of the intersections in Ketamine+ saline group((2.07±0.48), (1.67±0.72), P<0.05) increased. While the number of the intersections in Ketamine+ aucubin group was lower than that of Ketamine+ saline group ((1.20±0.78), (2.07±0.48), P<0.05). Conclusion:Aucubin administration can alleviate ADHD-like behaviors in offspring mice, and the mechanism may be associated with the inhibition of excessive astrocytic activation.
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Objective:To investigate the icariin on cognitive function and astrocytic pyroptosis in hemorrhagic shock resuscitation model mice.Methods:Forty-eight SPF grade C57BL/6 mice (male) were randomly divided into four groups ( n=12 in each group): Sham operation control group (Group C), hemorrhagic shock and resuscitation group (Group H), hemorrhagic shock and resuscitation plus icariin group (Group HI) and hemorrhagic shock resuscitation plus icariin and SSK1 group (Group HIS, SSK1 was a phosphorylation agonist of mitogen-activated protein kinase p38(p38MAPK). The mice in Group H, HI and HIS were subjected to hemorrhagic shock and resuscitation model by bleeding and retransfusion via left femoral vein; the mice in Group HI and HIS were administered with icariin (10 mg/kg) intragastrically for 7 days; the mice in Group C and H were administered with the same amount of normal saline containing dimethyl sulfoxide(DMSO). The mice in Group HIS were administered with SSK1 (0.5 mg/kg) intraperitoneally, but the mice in Group C, H and HI were only administered with the same amount of normal saline containing DMSO.At 15 days after resuscitation, novel objective recognition test and fear conditioning test were used to assess cognitive dysfunction of mice.Microtubule-associated protein 2(MAP2), a specific marker protein of neurons reflecting astrocytic pyroptosis in the hippocampus of mice, were detected by immunofluorescence assay so as to assess neuronal injury and astrocytic pyroptosis.The levels of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the hippocampus were evaluated by Western blot.SPSS 21.0 software was used for data analysis, multiple samples among groups were compared by one-way ANOVA, and SNK- q test was used for further pairwise comparison. Results:The results of new object recognition test showed that the difference of new object recognition index among the four groups was statistically significant ( F=50.75, P<0.05). The new object recognition indexes in H group(22.7±6.9), HI group(40.1±7.0) and HIS group (22.5±7.5) were significantly lower than that in C group (58.5±11.2). The index in HI group was higher than that in H group, while the index in HIS group was lower than that in HI group (all P<0.05). The results of the fear conditioning test showed that there was a statistically significant difference in the percentage of freezing time among the four groups of mice ( F=60.54, P<0.05). And the percentage of freezing time in H group((21.8±5.0)%), HI group ((38.4±7.4) %)and HIS group((21.3±4.2)%)were lower than that in C group((49.1±7.0)%), which in HI group was higher than that in H group ( P<0.05)and which in HIS group was lower than that in HI group(all P<0.05). The results of immunofluorescence showed that there were significant decreases of MAP2 intensity ((35.3±9.3)%, (63.3±6.1)%, (28.7±10.3)%) but increases of pyroptotic astrocytes ((24.5±4.2)%, (9.3±1.5)%, (22.1±3.3)%) in the H, HI and HIS groups compared with those of C group ((106.7±19.7) %, (3.4±2.0)%). There was an increase of MAP2 intensity but a decrease of pyroptotic astrocytes in the HI group compared with those in H group, and there was a decrease of MAP2 intensity but an increase of pyroptotic astrocytes in the HIS group compared with those of HI group (all P<0.05). The Western blot results showed that there were significant increases of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the H, HI and HIS groups compared with C group, there were decreases of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the HI group compared with H group, and there were increases of IL-1β, IL-18, the ratio of phosphorylated p38MAPK to total p38MAPK in the HIS group compared with those in HI group (all P<0.05). Conclusion:Icariin alleviates hemorrhage shock and resuscitation-induced cognitive dysfunction and astrocytic pyroptosis in mice, and the mechanism may be associated with inhibition of phosphorylated p38MAPK.
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Objective:To analyze the laboratory tests and clinical characteristics of patients with lupus anticoagulant-positive cerebral infarction.Methods:A retrospective analysis of 216 patients with cerebral infarction hospitalized in Beijing Tiantan Hospital from January 2016 to October 2021 was performed, and the patients were divided into LA-positive cerebral infarction group (168 cases) and LA-negative cerebral infarction group (48 cases) according to the detection of lupus anticoagulant (LA) in cerebral infarction patients, and the laboratory test data between the two groups were compared, and the risk factors related to cerebral infarction, including body mass index (BMI), smoking history, drinking history, hypertension, hyperlipidemia, diabetes history, were included for comparative analysis. LA was performed using the silica clotting time (SCT) method and the modified diluted russell viper venom time (dRVVT) method, respectively. The dRVVT method was used to detect LA. The LA-positive cerebral infarction group was divided into three subgroups according to the positive detection, namely, the dRVVT single-positive group (110 cases), the SCT single-positive group (40 cases) and the double-positive group (18 cases), and the comparison of laboratory indices between different subgroups was performed.The measurement data of normal distribution between the two groups were compared by independent sample t-test, and the mean between multiple groups was compared by ANOVA; The rank sum test was used to compare the median between the measurement data groups that did not conform to the normal distribution, and the χ 2 test was used to compare the counting data groups. Results:The levels of antithrombin Ⅲ and protein C of the LA-positive group ((102.85±14.39)% and (108.52±22.62)%) were all lower than those of the LA-negative group ((110.16±11.10)% and (116.34±18.14)%), the difference was statistically significant ( t values were 3.25, 2.20, P values were 0.001, 0.029, respectively). The levels of fibrinogen, homocysteine, high-sensitivity C-reactive protein (hs-CRP), erythrocyte sedimentation rate, white blood cells and neutrophil were (3.43(3.07,4.03) g/L), (17.92(14.07,23.71) μmol/L), (6.97(2.33,11.46) mg/L), (15.00(6.75,29.00) mm/h), (8.61(6.72,10.86)×10 9/L) and (5.81(4.39,7.91)×10 9/L), all were higher than those in the LA-negative group with values of (3.14(2.68, 3.62) g/L), (14.62(12.49, 18.41) μmol/L), (3.18(2.09,4.32) mg/L), (9.50(3.75,19.00) mm/h), (7.20(6.22,8.33)×10 9/L) and (4.47(4.02,5.57)×10 9/L), and the differences were statistically significant ( Z values were 2.77, 2.89, 3.32, 2.45, 3.15 and 3.76, P values were 0.006、0.004、0.001, 0.014, 0.002 and <0.001, respectively). There were no significant differences in age, gender, BMI, personal history, past medical history and other laboratory indicators between the two groups (all P>0.05). Comparison among different subgroups in LA positive group showed that D dimer and hs-CRP levels in double-positive group were 0.58(0.50,0.84) mg/L and 7.77(5.94,21.61) mg/L, higher than those in SCT single-positive group with values of 0.45(0.32,0.56) mg/L and 2.98(1.09,6.07) mg/L, and protein S level of double-positive group (97.36±25.45)% was lower than that in SCT single-positive group (114.85±22.74)%, the differences were statistically significant (all P<0.05). D dimer, prothrombin time, hs-CRP and neutrophil levels in dRVVT single-positive group were (0.58(0.50,0.84) mg/L), (11.40(11.10,12.10) s), (6.97(4.07,11.97) mg/L) and (5.83(4.51,8.27)×10 9/L), which were higher than those in SCT single-positive group with values of (0.45(0.32,0.56) mg/L), (11.15(10.70,11.43) s), (2.98(1.09,6.07) mg/L) and (5.08(3.92,6.07)×10 9/L), the difference was statistically significant (all P<0.05). Protein C and triglyceride levels were ((105.65±20.62)%) and (1.38(1.05, 1.75) mmol/L) in dRVVT single-positive group, which were lower than those in SCT single-positive group with values of ((117.05±20.86)% and 1.60(1.29,2.36) mmol/L), the differences were statistically significant (all P<0.05). Conclusion:There were significant differences between LA positive and LA negative cerebral infarction patients in laboratory examination. In LA positive cerebral infarction patients, the levels of fibrinogen, homocysteine, hs-CRP, white blood cells, neutrophil and erythrocyte sedimentation rate were higher, while the levels of anticoagulant protein antithrombin Ⅲ and protein C were lower. It is of great significance to pay close attention to the level and change of laboratory related risk factors in patients with LA positive cerebral infarction and give early intervention and treatment for the prevention of the occurrence and recurrence of cerebral infarction.
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In order to adapt to the development of higher medical education, it is imperative to carry out a student-centered teaching reform of pathophysiology guided by the cultivation of "clinical competence". The main contents of this teaching reform of pathophysiology include: the teaching content layout has been reconstructed to promote classroom teaching with clinical manifestation, new media means such as blackboard online teaching system, WeChat public platform and Chaoxing virtual classroom have been applied to assist students in their learning, and students are encouraged to conduct literature retrieval and reading, and then make presentation PPT for a literature of interest and report to all students to cultivate students' independent learning and sustainable development ability via flipped classroom. The practice shows that the student-centered teaching reform of pathophysiology guided by the cultivation of "clinical competence" has gained successful results.
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Objective:To explore the safety and feasibility of optimized pathological evaluation system of donor's kidney and modified surgery during adult dual kidney transplantation(DKT)and evaluate its effectiveness to provide more alternative protocols for kidney transplantation from extended criteria donors.Methods:DKT was performed in 10 recipients using the same protocol from June 2019 to May 2021.And retrospective reviewing was performed for clinical data, including characteristics of donors and recipients, optimized pathological evaluation system, modified surgery, treatment regimens, complications and follow-ups.Results:There were 8 male and 2 female donors with an age of(57.9±12.8)years and BMI(24.1±4.1)kg/m 2.The percentage of DCD was 70% and DBD 30%.The serum creatinine before procurement was 107.6(93.3-163.5)μmol/l.Zero-point puncture biopsy was performed for both kidneys and optimized pathological evaluation system was implemented(Banff criteria & Remuzzi score). The pathological results indicated that glomerular sclerosis for left and right kidneys were 2.0(1.5-2.0)and 1.5(1.0-2.0). And Remuzzi score for left and right kidneys were(4.4±1.2)and(3.6±1.5)points respectively.All recipients were male with an age of(43.1±9.0)years and BMI(22.2±1.9)kg/m 2.All PRAs were negative pre-operation.Modified surgery was performed in all recipients(two kidneys were implanted outside iliac vessels without patch and artery of superior kidney was anastomosed to internal iliac artery). Operative duration was(195±54.3)min and serum creatinine before discharge 125.0(102.0-199.0)μmol/L.Renal dynamic scintigraphy indicated that glomerular filtration rate was(30.0±8.2)ml/min for left kidney and(29.2±13.9)ml/min for right kidney.MRA results indicated that morphologies of renal arteries and veins were regular.The time between operation and discharge was(22.4±4.7)days.Compared with SKT, serum creatinine before discharge of DKT was lower and DGF incidence of DKT was higher without statistical significance.The time between operation and discharge was longer for DKT than that for SKT( P<0.05). The complications consisted of 20% donor derived infection(DDI)and 50% DGF.And there was no surgical complication associated with vessels and ureter.Renal function remained stable during 6-month follow-ups. Conclusions:Optimized pathological evaluation system of donor's kidney and modified surgery during adult dual kidney transplantation are both safe and feasible.The postoperative function of transplanted dual kidney is successfully restored.However, long-term follow-ups are required for evaluating its effectiveness.
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Objective:To analyze the clinical characteristics and risk factors of ischemic stroke in young adults.Methods:A retrospective analysis was conducted on 80 ischemic stroke patients (age ≤45 years) admitted to Beijing Tiantan Hospital from March 2019 to October 2019 as the young stroke group, and 117 ischemic stroke patients (age >45 years) hospitalized during the same period as the middle-aged and elderly stroke group. The blood test indexes of the two groups were compared, and the risk factors related to stroke, including smoking history, drinking history, hypertension, hyperlipidemia and diabetes history, were compared and analyzed. Two sets of independent sample t-test, Mann-Whitney U-test or χ2 test were used to compare the above indicators of patients in the two groups. Results:The activated partial prothrombin time, protein S, uric acid, homocysteine and D-dimer levels in middle-aged and elderly stroke group were (29.73±3.40) s, (105.58±27.23) %, (297.29±85.99) μmol/L, (17.58±14.45) μmol/L and (2.75±3.08) mg/L, respectively. Compared with the middle-aged and elderly stroke group, the young stroke group had higher activated partial thrombin time (31.51±6.75) s, protein S (115.20±26.97) %, uric acid (326.82±93.51) μmol/L, homocysteine (22.63±16.98) μmol/L and lower D dimer level of (1.19±2.88) mg/L compared with the elder group, the difference between the two groups was statistically significant ( t values were 2.17, 2.01, 2.20, 2.14 and 2.13, respectively, P values were 0.032, 0.046, 0.029, 0.039 and 0.034, respectively). The positive rate of lupus anticoagulant in young stroke group was 12.5% (4/32), which was higher than 1.8% (1/57) in middle-aged and elderly stroke group, and there was significant difference between the two groups (χ 2=4.46, P=0.035). The proportions of smoking and drinking in young stroke group were 63.8% (51/80) and 62.5% (50/80), respectively, which were higher than 49.6% (58/117) and 47.9% (56/117) in middle-aged and elderly stroke group, and there was significant difference between the two groups (χ 2 values were 3.86 and 4.09; P values were 0.04 and 0.04). The proportion of hypertension and diabetes in young stroke group was 48.8% (39/80) and 17.5%(14/80), respectively, which were lower than 63.2%(74/117) and 30.8%(36/117) in middle-aged and elderly stroke group, and there was significant difference between the two groups (χ 2 values were 4.08 and 4.56; P values were 0.043 and 0.033). According to the levels of uric acid and homocysteine, young stroke was divided into different subgroups and compared.The creatinine level of high uric acid group (≥416 μmol/L) was (90.08±28.46) mmol/L, which was higher than that of normal uric acid group (<416 μmol/L) of (63.37±22.2) mmol/L. There was significant difference between the two groups ( t value was 2.23, P value was 0.046). The levels of fibrinogen and creatinine in high homocysteine group (≥15 μmol/L) were (3.27±1.09) g/L and (72.13±28.69) mmol/L, respectively which were significantly higher than those in normal homocysteine group (<15 μmol/L) of (2.78±0.67) g/L and (58.92±12.08) mmol/L, There was significant difference between the two groups (the t values were 2.32 and 2.51; P values were 0.023 and 0.014). Conclusions:Compared with middle-aged and elderly stroke, young ischemic stroke has higher levels of prothrombin time, protein S, uric acid and homocysteine, lower levels of D dimer and higher positive rate of lupus anticoagulant. At the same time, the proportion of smoking and drinking was higher in young stroke group, but the proportion of hypertension and diabetes was relatively lower.
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Objective:To investigate the effect of activation of microglia in prefrontal cortex on long-term spatial memory in post-stroke depression mice.Methods:Forty-eight male C57BL/6 mice were divided into sham operation group, stroke group, post-stroke depression group and depression group according to the random number table method with 12 in each group, and 36 mice were divided into solvent group, enrofloxacin group and minocycline group according to the random number table method with 12 in each group.Middle cerebral artery occlusion (MCAO) was use to establish the stroke model, and forced swimming was used to establish the depression model.The post-stroke depression model mice were received MCAO first and then received forced swimming on the 4th day after stroke to establish the model.Mice in enrofloxacin group and minocycline group were treated with enrofloxacin and minocycline injection once a clay for 14 days from the 5th day after stroke, respectively.Forced swimming test and sugar water preference test were used to evaluate the depression of mice in each group, Morris water maze test was used to detect the spatial memory function of mice in each group, and Nissl staining and immunofluorescence staining were used to detect the neuronal function and the number and type of microglia activation.The expression of inflammatory cytokines IL-6 and IL-1β were detected by Western blot.GraphPad Prism 8.0.1 statistical software was used for statistical analysis.The single factor variance analysis was used to compare the difference among multiple groups, and pairwise comparison was performed with SNK- q test. Results:(1) There were statistically significant differences in depression, learning and memory, neuron damage, activation of microglia, inflammatory factors and other indicators in sham operation group, stroke group, post-stroke depression group and depression group ( F=43.58-255.70, all P<0.05). Compared with stroke group, post-stroke depression group had longer floating immobility time ((222.70±29.12) s, (79.25±46.78) s, P<0.05), the preference rate of sugar water was significantly lower ( (49.44±6.19) %, (84.49±4.73) %, P<0.05), and the average value of platform approach after correction was higher((125.00±9.95) mm, (96.79±12.57) mm, P<0.05), Nissl bodies expression was lower ((53.50±15.78) cells /mm 2, (85.67±17.52) cells /mm 2, P<0.05), NeuN positive expression rate was lower ((29.78±3.70) %, (45.73±4.51) %, P<0.05), the percent of M1 microglia expression was significantly higher ((75.55±8.84) %, (58.19±5.69) %, P<0.05), the percent of M2 microglia expression was lower ((43.46±5.11)%, (57.14±5.40)%, P<0.05), and the expression levels of IL-6 ((1.14±0.03), (0.94±0.05), P<0.05) and IL-1β((1.17±0.03), (0.56±0.04), P<0.05) were significantly higher.(2) Depression, learning and memory, neuron injury, activation of microglia, inflammatory factors and other indicators of mice in solvent group, enrofloxacin group and minocycline group were significantly different ( F=7.13-94.35, all P<0.05). Compared with enrofloxacin group, mice in minocycline group had shorter floating immobility time ((169.30±13.04) s, (224.30±22.60) s, P<0.05) and higher sugar water preference rate ((62.81±7.75) %, (47.71±8.11) %, P<0.05), the mean value of platform approach estimation after water maze correction was lower ((97.66±14.56) mm, (120.20±12.08) mm, P<0.05), and the expression level of Nissl bodies was higher ((80.17±10.55) cells /mm 2, (52.00±8.94) cells /mm 2, P<0.05), NeuN expression rate was high ((45.04±3.62) %, (28.88±4.50) %, P<0.05), Iba-1 expression was lower ((97.33±10.67) cells/mm 2, (112.50±6.54)cells/mm 2, P<0.05), the percent of M1 microglia expression was lower ((54.43±5.22) %, (73.82±6.88) %, P<0.05), and the percent of M2 microglia expression was significantly higher ((51.86±6.22) %, (36.30±5.72) %, P<0.05). The expression levels of IL-6 ((0.75±0.06), (1.21±0.07), P<0.05) and IL-1β ((0.61±0.06) (1.09±0.09), P<0.05) were lower. Conclusion:The long-term spatial memory impairment of post-stroke depression mice is aggravated, which is related to the neuron damage caused by increased activation of M1 microglia in PFC area.Inhibition of M1 microglia by minocycline can effectively improve the spatial memory ability of mice.
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Objective@#To explore dynamics of parenting styles of adolescents from 1999 to 2019 from the perspective of intergenerational conflict, to provide support for family education and adolescent healthy development.@*Methods@#Using a multistage stratified cluster random sampling method, the unified questionnaire was administered to 2 590 students in the same sampling junior and senior high schools in 1999, 2009, and 2019 using the Egna Minnen av Barndoms Uppfostran own memories of parental rearing practices in childhood(EMBU).@*Results@#Overall there were differences in the nine factors of parenting styles across generations ( F = 12.07-72.52, P <0.01), with decreasing ratings of warmth and understanding of father and mother (F1, M1), over interference of father (F3) over generations(F1:46.72±9.41, 45.87±11.33, 43.61±11.27; M1:51.56±9.38, 51.03±11.59, 46.23± 12.27 ; F3:19.03±4.00, 18.29±4.32, 17.95±4.51), and all other parenting styles rated higher in 2019 than in 2009 and 1999(except for the over protection and over interference of mother, and punishment, firm control of mother). Parenting styles across generations (except for the rejection and denial of father among girls) showed gender difference.The overall gender trend coincided with the total population trend. Parenting styles across generations varied significantly among middle and high school students( F =3.92-47.27, P <0.05 ), changes in F1 and F3 factors coincided with the overall decreasing trend. Factor analysis showed that parenting styles could be classified into two dimensions, with varied factor loading across generation.@*Conclusion@#Intergenerational decreases in parental emotional warmth and paternal interfering are observed in a sex and grade specific manner. Based on the diversity of needs and population differentiation, optimal intervention for comprehensive health development of adolescents are in great need to keep pace with the times and promoting the high quality development of adolescents.
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Objective:To evaluate the role of stimulator of interferon genes (STING) signaling pathway in carbon monoxide (CO)-releasing molecule-3 (CORM-3)-induced reduction of hepatocyte pyroptosis and apoptosis in a rat model of hepatic ischemia-reperfusion.Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 9-11 weeks, weighing 320-380 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (S group), ischemia-reperfusion group (IR group), CORM-3 group (C group) and STING agonist ADU-S100 group (A group).Hepatic ischemia-reperfusion injury models were developed by reversible ligation of left middle hepatic artery, portal vein and bile duct branches for 45 min, followed by reperfusion in anesthetized animals in IR, C and A groups.In group C, CORM-3 4 mg/kg was injected into the femoral vein immediately after reperfusion.The equal volume of normal saline containing dimethyl sulfoxide was injected into the femoral vein in S, IR and A groups.At 1.5 h after injection into the femoral vein, ADU-S100 10 mg/kg was intraperitoneally injected in A group, and the equal volume of normal saline was given instead in S, IR and C groups.The serum alanine transaminase (ALT) and aspartate transaminase (AST) concentrations were determined at 3 h of reperfusion.The rats were sacrificed at 12 h of reperfusion, and liver tissues were collected for determination of the content of CO (by colorimetry), expression of interleukin-1beta (IL-1β), IL-18, Bcl-2, Bax, interferon regulatory factor 3 (IRF3), phosphorylated IRF3 (p-IRF3), STING, NOD-like receptor protein 3 (NLRP3), aspirin D (GSDMD) and activated caspase-1 (by Western blot), and pyroptosis and apoptosis rates of hepatocytes (by immunofluorescence staining).The liver injury was scored. Results:Compared with group S, the serum ALT and AST concentrations, liver injury score, CO content, and pyroptosis and apoptosis rates of hepatocytes were significantly increased, and the expression of IL-1β, IL-18, p-IRF3, STING, NLRP3, GSDMD and activated caspase-1 was up-regulated, and the Bcl-2/Bax ratio was decreased in group IR ( P<0.05).Compared with group IR, the serum ALT and AST concentrations, liver injury score, and pyroptosis and apoptosis rates of hepatocytes were significantly decreased, the CO content was increased, the expression of IL-1β, IL-18, p-IRF3, STING, NLRP3, GSDMD and activated caspase-1 was down-regulated, and the Bcl-2/Bax ratio was increased in group C ( P<0.05).Compared with group C, the serum ALT and AST concentrations, liver injury score, and pyroptosis and apoptosis rates of hepatocytes were significantly increased, the CO content was decreased, the expression of IL-1β, IL-18, p-IRF3, STING, NLRP3, GSDMD and activated caspase-1 was up-regulated, and the Bcl-2/Bax ratio was decreased in group A ( P<0.05). Conclusions:The mechanism by which CORM-3 attenuates hepatocyte pyroptosis and apoptosis may be related to the inhibition of activation of STING signaling pathway in a rat model of hepatic ischemia-reperfusion.
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Objective:To explore the early warning value of carbapenem-resistant Klebsiella pneumoniae (CRKP) positivity in liver transplantation recipients with rectal swabs, examine the risk factors of CRKP bloodstream infection and provide the relevant treatments.Methods:From June 2018 to December 2019 in Organ Transplantation Research Institute Affiliated Tongji Hospital Tongji Medical College Huazhong University of Science & Technology, 148 cases of liver transplantation recipients with positive CRKP rectal swabbing were recruited. Clinical data were retrospectively analyzed. And the risk factors of CRKP bloodstream infections were examined for intervention and non-intervention groups to observe the effect of interventions of CRKP bloodstream infections.Results:Among them, 23 cases (15.5%) were positive for CRKP and 5 cases (21.7%) were infected with CRKP bloodstream. Rectal swab culture was negative in 125 cases and no bloodstream infection occurred. Long-term use of broad-spectrum antimicrobial agents, severe basic diseases (severe hepatitis), postoperative delayed graft liver function recovery, acute renal failure requiring renal replacement therapy (RRT) and postoperative anti-thymocyte globulin (ATG) induction were risk factors. In intervention group, there were 2 cases (11.1%) of 18 patients with positive CRKP in rectal swab culture in late stage. Among 5 CRKP-positive recipients without intervention, 3 cases (60%) developed later CRKP bloodstream infection. The incidence of bloodstream infection was significantly lower in intervention group than that in non-intervention group ( P<0.05). Conclusions:Rectal swab culture for liver transplantation recipients provides early warning for CRKP bloodstream infection. Interventions for CRKP positive high-risk recipients with rectal swab culture may reduce the occurrence of CRKP bloodstream infection and lower the risk of CRKP bloodstream infection in liver transplantation recipients.
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Objective:To investigate the value of two-dimensional ultrasound combined with shear wave elastography (SWE) in assessing perineal body characteristics in patients with stress urinary incontinence(SUI).Methods:Sixty-four patients with stress urinary incontinence from December 2019 to August 2020 in the Affiliated Hospital of Qingdao University were selected as the SUI group, in addition, seventy healthy females at the same time were selected as the control group. The clinical information data were collected, two-dimensional ultrasound and SWE examination were performed to obtain the perineal body length, height, perimeter, area and the maximum(Emax) and the mean(Emean) modulus of elasticity both at the resting and maximum Valsalva conditions, ROC curve was plotted to evaluate the effectiveness of elastic modulus in evaluating perineal body in SUI.Results:In the resting state, the length, height, perimeter and area of perineal body in the SUI group were not significantly different from those of the control group ( P=0.590, 0.291, 0.082, 0.063). At the maximum Valsalva, the perineal body length, height, perimeter and area in the SUI group were significantly different from those of the control group ( P=0.005, 0.010, 0.001, 0.008). In the resting, the Emax and Emean values of perineal body elastic modulus in the SUI group were higher than those in the control group, but the difference was not statistically significant [(36.61±9.81)kPa vs (34.66±10.38)kPa, (27.43±9.78)kPa vs (26.97±8.85)kPa, all P>0.05]. At the maximum Valsalva, the Emax and Emean of the SUI group were significantly smaller than those of the control group, with statistically significant difference[(47.73±8.03)kPa vs (58.06±10.02)kPa, (35.78±7.89)kPa vs (44.33±9.62)kPa, all P<0.001]. The area under ROC curve of Emax and Emean at the maximum Valsalva was 0.738 and 0.647 respectively, the Emax was better than the Emean in assessment of the characteristics of perineal body in SUI patients. Conclusions:Two-dimensional ultrasound combined with SWE technique can quantitatively evaluate the characteristics of perineal body and provide an important imaging method for the diagnosis of SUI.
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Objective:To analyze the clinical characteristics and risk factors of clopidogrel resistance after intracranial arterial stent implantation in patients with ischemic cerebrovascular disease.Methods:Retrospective case-control study was used.A total of 360 patients with ischemic cerebrovascular disease hospitalized in Beijing Tiantan Hospital from January 2017 to December 2018 were selected.All patients received intracranial arterial stenting and received double anti-platelet drugs after operation.Patients were divided into clopidogrel resistance group and non-resistance group according to the inhibition rate of platelet aggregation measured by thromboelastography.Clinical data and laboratory indicators of patients in the two groups were compared using two independent sample T tests and Mann-Whitney U tests, and possible risk factors of clopidogrel resistance were analyzed by Logistic regression.Results:White blood cell (WBC) count, neutrophils count, lymphocyte count, cholesterol, LDL cholesterol, and serum homocysteine levels in the clopidogrel non-resistant group were (6.58±1.45)×10 9/L, (4.01±1.05)×10 9/L, 1.83(1.49, 2.23)×10 9/L, (3.63±0.85) mmol/L, 1.93(1.53, 2.31) mmol/L, and 14.3(11.80, 17.00) μmol/L, respectively.Compared with the clopidogrel non-resistant group, the WBC count, neutrophils count, lymphocyte count, cholesterol, LDL cholesterol and serum homocysteine levels in the clopidogrel resistant group were all higher (7.19±1.53) ×10 9/L, (4.40±1.05) ×10 9/L, 2.03(1.63, 2.58)×10 9/L, (4.02±0.99) mmol/L, 2.04(1.68, 2.78) mmol/L and 15.90(12.25, 22.20) μmol/L, respectively.The difference was statistically significant ( t=3.277, t=2.867, Z=2.457, t=3.409, Z=2.705 and Z=2.220, respectively; P value were 0.001, 0.004, 0.014, 0.001, 0.007 and 0.026, respectively). Logistic regression analysis showed that high serum homocysteine ( OR=1.031, 95% CI 1.006-1.057, P=0.015) was an independent risk factor for clopidogrel resistance. Conclusion:Patients with clopidogrel resistance have high leucocyte, blood lipid, homocysteine and other clinical characteristics.High serum homocysteine is an independent risk factor for clopidogrel resistance.
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Objective:To investigate the effects of Dickkopf-1(DKK1) on the proliferation of human lens epithelial cells (LECs) and its possible mechanism in order to search a new target for the treatment of posterior capsular opacification (PCO).Methods:Human LECs line (SRA 01/04 cells )were divided into Wnt3a overexpression group, DKK1 group and control group.Wnt3a gene expression vector was transfected into SRA 01/04 cells by liposome mediated transfection to establish a PCO model in the Wnt3a overexpression group, DKK1 of 100 μg/ml was added into the medium 48 hours after transfection of Wnt3a gene vector in the DKK1 group, and only pcDNA3-HA vector was transfected in the control group.The survival rate of SRA 01/04 cells was detected with a cell counting kit-8 (CCK-8) assay.The expression rate of proliferating cell nuclear antigen (PCNA) in the cells was detecteds by immunocytochemistry.The expression of β-catenin in the cells was detected and located by immunofluorescence.The expression of Wnt3a CyclinD1 and C-Myc were detected by Western blot assay.Results:The relative expression of Wnt3a protein in the control group was 0.49±0.07, which was significantly lower than that in the Wnt3a overexpression group (0.84±0.06) ( t=3.704, P=0.02). The survival rate in the Wnt3a overexpression group, DKK1 group and control group showed significant difference over time ( Fgroup=10.910, P<0.05; Ftime=6.041, P<0.05). The survival rate in the Wnt3a overexpression group was significantly increased in comparison with the control group and that in the DKK1 group was significantly reduced in comparison with the Wnt3a overexpression group(all at P<0.05). β-Catenin was expressed mainly in cytoplasm and cell nucleus in the Wnt3a overexpression group and only in cytoplasm in the DKK1 group. in the control group, β-catenin showed a weaked expression in the cytoplasm and nucleus in comparison with the Wnt3a overexpression group.The expression rates of PCNA protein were (9.4±1.4)%, (43.4±5.4)%, and (14.2±2.3)% in the control group, Wnt3a overexpression group and DKK1 group, respectively, with a significant difference among the groups ( F=28.250, P<0.05), and the expression rates of PCNA protein were significantly reduced in the control group and DKK1 group compared with the Wnt3a overexpression group (both at P<0.05). β-Catenin protein were expressed mainly in the cytoplasm and nucleus in the Wnt3a overexpression group and only in the cytoplasm in the control group.In the DKK1 group, the expression of β-catenin protein was weakened in the cytoplasm and nucleus in comparison with the Wnt3a overexpression group.The relative expressions of CyclinD1 were 0.64±0.07、0.84±0.03 and 0.55±0.10, C-Myc were 0.59±0.05、0.93±0.02 and 0.47±0.08 in the control group, Wnt3a overexpression group and the DKK1 group, respectively with significant differences among the groups ( F=20.580, 5.040, both at P<0.05). The relative expressions of CyclinD1 and C-Myc in the Wnt3a overexpression group were significantly higher than those in the DKK1 group and the control group (all at P<0.05). Conclusions:DKK1 inhibits activation of Wnt/β-catenin signaling pathway induced by Wnt3a overexpression in SRA01/04 cells, and down-regulation of downstream target proteins cyclin D1 and C-Mgc may be the biological mechanism of Dkk1 inhibiting human LECs proliferation.
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The procurement, preservation and transportation of the donor organs directly affect the clinical prognosis of the recipients. The establishment of process optimization and quality control standards of organ procurement, preservation and transportation contributes to improving the quality and utilization rate of donor organs and reducing the medical risk. According to Guide to the Quality and Safety of Organs for Transplantation (6th edition) proposed by European Union, the 11th chapter of organ procurement, preservation and transportation was interpreted and summarized in this article.
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Objective:To investigate the risk factors of Crohn′s disease (CD)-related gastrointestinal stenosis, and to summarize and analyze the corresponding treatments.Methods:From January 2010 to December 2018, 122 patients diagnosed with CD and hospitalized in the Seventh Medical Center, PLA General Hospital were selected including 72 patients in gastrointestinal stenosis group and 50 patients in non-gastrointestinal stenosis group. The gender, age of onset, course of disease, location of lesions involved (Montreal classification), disease activity, extraintestinal manifestations, application of therapeutic drugs, and complications were compared between the two groups. The treatment of CD patients with gastrointestinal stenosis was analyzed. Multivariate logistic regression was used to analyze the risk factors of CD patients with gastrointestinal stenosis. The independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:The age of onset of patients in gastrointestinal stenosis group was older than that in non-gastrointestinal stenosis group ((37.6±15.1) years old vs. (30.8±14.7) years old), and course of disease was longer than that of non-gastrointestinal stenosis group (72 months, 11 to 492 months vs. 45 months, 3 to 240 months); and the differences were statistically significant ( t=-2.044, Z=-2.770; P=0.018, 0.006). The proportion of patients with ileum involvement of the gastrointestinal stenosis group was lower than that of the non-gastrointestinal stenosis group (69.4%, 50/72 vs. 86.0%, 43/50), and the proportion of severe patients was higher than that of the non-gastrointestinal stenosis group (15.3%, 11/72 vs. 4.0%, 2/50); and the differences were statistically significant ( χ2=4.463 and 3.942, P=0.035 and 0.047). There were no significant differences in gender, use of therapeutic drugs, extraintestinal manifestations, application of therapeutic drugs or the incidence of complications between the patients of two groups (all P>0.05). The results of multivariate logistic regression showed that the age of onset and course of disease were risk factors of CD-related gastrointestinal stenosis ( β=0.028, odds ratio ( OR)=1.028, 95% confidence interval ( CI) 1.000 to 1.056, P=0.046; β=0.008, OR=1.008, 95% CI 1.002 to 1.015, P=0.013). Further stratified analysis revealed that the incidence rates of CD-related gastrointestinal stenosis in patients with age of onset over 40 years old and course of disease more than five years were higher than those of patients with age of onset less than 40 years old and course of disease less than five years (76.3%, 29/38 vs. 51.2%, 43/84; 68.4%, 39/57 vs. 50.8%, 33/65), and the differences were statistically significant ( OR=3.072, 95% CI 1.298 to 7.272, P=0.009; OR=2.101, 95% CI 1.002 to 4.406, P=0.048). Among the 72 CD patients with gastrointestinal stenosis, 15 cases (20.8%) were treated with medicine and nutrition, without endoscopic or surgical treatment. Fifty-two patients (72.2%) underwent surgical treatment, among them six patients (11.5%) received twice surgery, the interval between the two operations was 46 months (1 to 204 months), and eight patients (15.4%) had postoperative complications. Twenty-one patients (29.2%) were treated with endoscopic dilatation, and no complications occurred after surgery. Five patients (23.8%) underwent surgical treatment during the follow-up period. Conclusions:The age of onset over 40 years old and the course of disease more than five years are the risk factors of CD-related gastrointestinal stenosis. Individualized medical treatment is the basis for the treatment of CD-related gastrointestinal stenosis. Surgery is still the main treatment. The endoscopic treatment is safety and can delay or avoid surgery to a certain extent.
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Objective:To evaluate the effect of carbon monoxide-releasing molecule-3 (CORM-3) on blood transfusion-related acute lung injury in rats with traumatic brain injury (TBI).Methods:Seventy-two clean-grade healthy adult male Sprague-Dawley rats, weighing 300-350 g, were divided into 4 groups ( n=18 each) using the random number table method: sham operation group (group S), TBI group (T group), TBI plus 10 ml/kg plasma transfusion group (TP group), and TBI plus 10 ml/kg plasma transfusion plus CORM-3 group (TPC group). TBI was induced by dropping a 20-g weight from 20 cm height falling freely in anesthetized rats.Plasma 10 ml/kg was infused via the femoral vein after TBI in TP and TPC groups.The rats were sacrificed at 24 h after plasma transfusion, and lung tissues were obtained for determination of wet/dry weight (W/D) ratio, cell apoptosis, and expression of caspase-3, Bid, Bim and Puma (by Western blot). The lung injury score was calculated using the results of HE staining.Lung ultrasonography was performed for assessment of sonographic score, and the apoptosis rate was calculated by the TUNEL staining method. Results:Compared with S group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly increased, and the expression of activated caspase-3, Bid, Bim and Puma was up-regulated in the other three groups ( P<0.05). Compared with T group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly increased, and the expression of activated caspase-3, Bid, Bim and Puma was up-regulated in TP group ( P<0.05). Compared with TP group, the W/D ratio, lung injury score, sonographic score and apoptosis rate were significantly decreased, and the expression of activated caspase-3, Bid, Bim and Puma was down-regulated in TPC group ( P<0.05). Conclusion:CORM-3 can reduce acute lung injury related to blood transfusion in rats with TBI, and the mechanism may be related to inhibiting cell apoptosis in lung tissues.
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Objective:To evaluate the effect of exogenous carbon monoxide (CO) on cell apoptosis during acute renal injury induced by hemorrhagic shock and resuscitation (HSR) in rats.Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 9-10 weeks, weighing 350-400 g, were divided into 4 groups ( n=12 each) by a random number table method: sham operation group (S group), HSR group (H group), HSR plus CORM-3 group (HC group) and HSR plus iCORM-3 group (HiC group). Mean arterial pressure was maintained at 30-35 mmHg for 45 min by withdrawing blood from the femoral vein, and the shed blood was re-transfused within 15 min to reach the initial blood pressure for resuscitation.Normal saline was infused when necessary, and the model of HSR was established.CORM-3 4 mg/kg and iCORM-3 4 mg/kg were added during resuscitation in HC group and HiC group, respectively.Only femoral vein and artery puncture was performed in S group.Blood samples were obtained from the tail vein at 3 h after resuscitation for measurement of serum urea nitrogen (BUN) and creatinine (Scr) concentrations.Rats were sacrificed at 12 h after resuscitation, and renal tissues were obtained for determination of the expression of Bcl-2 and Bak protein and cleaved caspase-3 (by Western blot) and cell apoptosis (by TUNEL). The damage to the renal tubules was assessed by paller assay after HE staining.Bcl-2/Bak ratio and apoptosis rate were calculated. Results:Compared with group S, the serum BUN and Scr concentrations, paller scores, and apoptosis rate were significantly increased, Bcl-2/Bak ratio was decreased, and the expression of cleaved caspase-3 was up-regulated in H, HC and HiC groups ( P<0.05). Compared with group H, the serum BUN and Scr concentrations, paller scores, and apoptosis rate were significantly decreased, Bcl-2/Bak ratio was increased, and the expression of cleaved caspase-3 was down-regulated in group HC ( P<0.05). Compared with group HC, the serum BUN and Scr concentrations, paller scores, and apoptosis rate were significantly increased, Bcl-2/Bak ratio was decreased, and the expression of cleaved caspase-3 was up-regulated in group HiC ( P<0.05). There was no significant difference in the indexes mentioned above between group H and group HiC ( P>0.05). Conclusion:The mechanism by which exogenous CO improves acute kidney injury may be related to inhibiting cell apoptosis in a rat model of HSR.
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Objective:To evaluate the effect of selective inhibitor of caspase-1 VX-765 on cognitive function in a rat model of hemorrhage shock and resuscitation (HSR).Methods:Forty-eight clean-grade healthy male Sprague-Dawley rats, aged 9-10 weeks, weighing 350-400 g, were divided into 4 groups ( n=12 each) using a random number table method: sham operation group (S group), HSR group (H group), VX-765 group (V group), and solvent control group (C group). The rats in H, V and C groups were subjected to hemorrhage by bleeding from femoral vein to achieve mean arterial pressure of 25-35 mmHg which was maintained at this level for 60 min followed by resuscitation with shed blood within 15 min to restore blood pressure, and normal saline was infused when needed.VX-765 1 mg/kg and 0.4% polyethylene glycol 1 mg/kg were intravenously injected via the femoral vein immediately after the end of resuscitation in V and C groups, respectively.Six rats in each group were selected and sacrificed at 12 h after the end of resuscitation, and the cerebral cortex was removed for determination of neuronal pyroptosis (by immunofluorescence) and degree of cortical edema (using T2-weighted imaging). Cognitive function was measured by open field test on day 7 after resuscitation in the rest 6 rats in each group. Results:Compared with S group, the pyroptosis rate in cortical neurons at 12 h after resuscitation and degree of cortical edema were significantly increased, the distance in the central square and the number of standing on the back legs were decreased on day 7 after resuscitation, and the time spent in the central square was shortened in H, V and C groups ( P<0.05). Compared with H and C groups, the pyroptosis rate in cortical neurons at 12 h after resuscitation and degree of cortical edema were significantly decreased, the distance in the central square and the number of standing on the back legs were increased on day 7 after resuscitation, and the time spent in the central square was prolonged in V group ( P<0.05). Conclusion:VX-765 can improve the cognitive function, and the mechanism may be associated with inhibiting pyroptosis in cortical neurons in a rat model of HSR.
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Objective To explore the relationship between brain injury related symptom groups and acute stress disorder( ASD). Methods Totally 301 patients with brain injury hospitalized in the Affiliated Hospital of North China University of Technology from March 2017 to March 2018 were recruited. The pa-tients were assessed by Stanford Acute Stress Response Questionnaire ( SASRQ), Brain Trauma Subscale (BIS) and Pittsburgh Sleep Quality Index (PSQI). Results The prevalence of acute stress disorder pa-tients with brain injury was 46. 51% (140/301). The SASRQ score of all patients with brain injury was (70. 78±28. 00). Univariate analysis showed that there were significant differences in age (χ2= 4. 172,P=0. 041),personality type (χ2= 57. 281,P<0. 001),sleep quality ( χ2= 70. 785,P<0. 001),hospitalization frequency (χ2= 43. 547,P<0. 001),depression,obsessive-compulsive and hostile symptoms between pa-tients with and without ASD ( P<0. 05). The scores of depression and obsessive-compulsive symptoms were significantly correlated with SASRQ scores(P<0. 05). The scores of hostility symptoms were significantly cor-related with SASRQ scores except for patients who lost their temper uncontrollably ( r=0. 176-0. 698,P<0. 05). Personality ( OR=0. 243,95% CI=0. 142-0. 415),sleep disorders( OR=0. 072,95% CI=0. 029-0. 178),hospitalization times(OR=0. 129,95%CI=0. 054-0. 306),depressive symptoms( OR=1. 260,95% CI=1. 061-1. 497),obsessive-compulsive symptoms(OR=1. 419,95%CI=1. 239-1. 625) and hostile symp-toms(OR=1. 253,95%CI=1. 058-1. 482) were the influencing factors of ASD in patients with brain injury (P<0. 05). Conclusion The incidence of ASD is higher in patients with brain injury. Personality type,hos-pitalization,sleep quality,depressive symptoms,obsessive-compulsive symptoms and hostile symptoms are the influencing factors,which should be paid more attention.
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Objective To identify the risk factors for postoperative short-term cognitive dysfunction ( STCD) in the patients undergoing carotid endarterectomy. Methods A total of 158 patients with carotid artery stenosis, aged≥18 yr, without preoperative cognitive dysfunction, undergoing carotid endarterecto-my from January 1, 2014 to May 30, 2017 in our hospital, were selected. Factors including age, sex, height, weight, education, presurgical complications ( hypertension, type Ⅱ diabetes mellitus, myocar-dial infarction and brain infarction) , degree of internal carotid artery stenosis, preoperative mean arterial pressure ( MAP ) , amount of dexmedetomidine, duration of operation, time of carotid artery occlusion, MAP during occlusion period, intraoperative volume of fluid infused ( crystalloid solution and colloid solu-tion), urine volume, blood loss, significant change in electroencephalogram (EEG) and visual analogue scale scores at 24 h after surgery were collected. Mini-Mental State Examination was used to evaluate the cognitive function at 24 h after surgery. The patients were divided into STCD group and non-STCD group ac-cording to whether or not patients developed postoperative STCD. The risk factors of which P values were less than 0. 05 would enter the bivariate logistic regression analysis to stratify the risk factors for postopera-tive STCD. Results Forty-nine patients developed postoperative STCD ( 31. 0％) . The intraoperative a-mount of colloid solution infused＜5 ml/kg, increase in MAP＜20％ of the baseline value during the occlu-sion period, and significant change in EEG during the occlusion period were independent risk factors for postoperative STCD ( P＜0. 05) . Conclusion The intraoperative amount of colloid solution infused＜5 ml/kg, increase in MAP＜20％ of the baseline value during the occlusion period, and significant change in EEG during the occlusion period are independent risk factors for postoperative STCD in the patients undergo-ing carotid endarterectomy.