Aim To investigate the toxicity of isopsor-alen in HepG2 cells and its effects on bile acid, bile acid synthesis and transport. Methods Cell viability was evaluated by MTT assay and bile acid was deter-mined inside HepG2 cells, with exposure to various isopsoralen for 24h. The mRNA transcription of BSEP, MRP2, MRP3, NTCP, OATP2, OSTα, CYP7A1, CYP27 A1 , FXR and PXR were assessed by real-time PCR. Results The cell viability was decreased dose-dependently with isopsoralen in HepG2 cells, and IC50 was 118. 1μmol·L-1 exposure to isopsoralen for 24h. Bile acid inside cells significantly increased with 100 and 400 μmol · L-1 isopsoralen. Isopsoralen caused the down-regulation of MRP2 , MRP3 , CYP7 A1 mRNA at 25 μmol · L-1 . Beside these, the up-regulation of OATP2,OSTα,CYP27A1,FXR,PXR with 100 μmol· L-1 isopsoralen, but there was no significant change of BSEP and NTCP. Conclusion The results show that isopsoralen induces bile acid accumulation and cytotox-icity which may be associated with the down-regulation of MRP2, MRP3 in HepG2 cells.

OBJECTIVE:To establish UPLC fingerprint of Glycyrrhiza uralensis. METHODS:UPLC method was adopted. The determination was performed on Waters CORTECS UPLC C18column with mobile phase consisted of acetonitrile-0.1% formic acid(gradient elution)at the flow rate of 0.3 mL/min. The detection wavelength was set at 254 nm,and column temperature was 30 ℃. The sample size was 1 μ L. Using glycyrrhizic acid as control,UPLC chromatograms of 27 batches of sample were determined. Similarity evaluation was conducted by using TCM Chromatogram Fingerprint Similarity Evaluation System(2004 A edition)to determine common peak and conduct cluster analysis of 27 batches of samples. RESULTS:There were 20 common peaks in UPLC chromatograms of 27 batches of samples. The similarity degree of S2,S4,S19,S21,S22,S24 were less than 0.90, the others samples were more than 0.90.After validation,UPLC chromatograms of 21 batches of batches of samples were in good agreement with control fingerprint. 27 batches of samples were clustered into 3 categories,in which S24 was categoryⅠ;S2,S4, S12,S19,S21,S22 were categoryⅡ;other were categoryⅢ. CONCLUSIONS:Established fingerprint can provide reference for quality evaluation of G.uralensis.

Objective:To evaluate the value of cerebrospinal fluid (CSF) nerve injury-related proteins levels in predicting postoperative delirium (POD) in patients.Methods:A total of 1 000 patients of both sexes, aged 40-90 yr, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with Mini-Mental State Examination score>24 at 1 day before operation, undergoing elective knee/hip arthroplasty under spinal-epidural anesthesia , were enrolled in this study.Cubital venous blood samples were drawn before anesthesia for detection of the concentrations of plasma total cholesterol, high-density lipoprotein, low-density lipoprotein and triglyceride.CSF 2ml was extracted after successful spinal-epidural anesthesia puncture for measurement of concentrations of α-synuclein (α-syn), β-amyloid protein 1-40 (Aβ 1-40), Aβ 1-42, total-Tau (t-Tau), phosphorylated Tau (p-Tau), progranulin (PGRN) and soluble myeloid cell triggering receptor 2 (sTREM2) (by enzyme-linked immunosorbent assay). The Confusion Assessment Method was used at 1, 3 and 7 days after surgery to evaluate the occurrence of POD.The patients were divided into POD group and non-POD group according to whether POD occurred after operation.Logistic regression analysis was used to analyze the variables of which P values were less than 0.05 to analyze the risk factors for POD.The receiver operating characteristic (ROC) curve was drawn and area (AUC) under the curve was calculated to evaluate the accuracy of the related risk factors in predicting POD. Results:A total of 964 patients were enrolled in the study, and 108 patients were diagnosed with POD, and the incidence was 11.2%.The results of logistic regression analysis found that age and and increased α-syn in CSF concentration were risk factors for POD, and decreased PGRN in CSF concentration and Aβ 1-42/p-Tau in CSF were the protective factors for POD ( P<0.05). ROC curve analysis showed that α-syn (AUC 0.69, 95% confidence interval (CI) 0.634-0.748, sensitivity 57.41%, specificity 82.10%, Youden Index 0.3951), PGRN in CSF concentration (AUC 0.695, 95%CI 0.637-0.750, sensitivity 59.26%, specificity 80.86%, Youden Index 0.4012) and Aβ 1-42/p-Tau in CSF (AUC 0.635, 95%CI 0.574-0.692, sensitivity 93.52%, specificity 30.25%, Youden Index 0.2377) could predict the occurrence of POD. Conclusion:PGRN, α-syn concentration and Aβ 1-42/p-Tau in CSF can predict the occurrence of POD in patients.

Objective:To investigate the relationship between preoperative subjective cognitive decline (SCD) and postoperative delirium (POD) in elderly patients.Methods:A total of 292 elderly patients of both sexes, aged 65-90 yr, weighing 50-90 kg, of American Society of Anesthesiologists physical status Ⅰ-Ⅱ, with Mini-Mental State Examination (MMSE) score>23 and Montreal Cognitive Assessment (MoCA) score > 26 at 1 day before operation, underwent total knee/hip arthroplasty under combined spinal-epidural block in our hospital from January to December 2020, were collected.The development of SCD was evaluated using subjective cognitive decline scale at 1 day before operation.Cerebrospinal fluid (CSF) was extracted after successful spinal-epidural anesthesia puncture, the concentrations of β-amyloid protein 40 (Aβ 40), Aβ 42, total tau (t-tau) and phosphorylated tau (p-tau) were determined by enzyme-linked immunosorbent assay.The incidence of POD was evaluated using confusion assessment method during post-anesthesia care unit and at 1-7 days after operation (or before discharge). Patients were divided into POD group and non-POD group according to whether POD occurred within 7 days after operation.The risk factors of which P values were less than 0.05 would enter the logistic regression analysis to stratify the risk factor for incidence of POD. Results:A total of 205 patients were enrolled and 53 patients developed POD (25.8%). The results of logistic regression analysis showed that preoperative SCD, and increased CSF p-tau and t-tau concentrations were risk factors for POD of elderly patients, and increased CSF Aβ 42 concentration and Aβ 40/p-tau, Aβ 40/t-tau, Aβ 42/p-tau and Aβ 42/t-tau were the protective factors for POD in elderly patients ( P<0.05). After correction of the confounding factors such as age, sex, body weight, education, the history of smoking and drinking, hypertension, diabetes and coronary heart disease, family history of dementia, Pittsburgh sleep quality index (PSQI), MMSE and MoCA score at 1 day before operation, duration of surgery, duration of anesthesia, intraoperative volume of infusion and blood loss and postoperative pain score, SCD, and increased CSF p-tau and t-tau concentrations were still the risk factors for POD in elderly patients, and increased CSF Aβ 42 concentration and Aβ 40/p-tau, Aβ 40/t-tau, Aβ 42/p-tau and Aβ 42/t-tau were still the protective factors for POD in elderly patients ( P<0.05). Conclusion:Preoperative SCD is the risk factor for POD in elderly patients.

Objective:To evaluate the changes in proteome in hippocampus and bioinformatics analysis in mice with perioperative neurocognitive disorders (PND).Methods:Clean-grade healthy male C57BL/6 mice, aged 15 months, weighing 30-35 g, were divided into 2 groups ( n=9 each) using a random number table method: control group (group C) and group PND.The model of PND was established by performing open tibial fracture with intramedullary fixation under isoflurane anesthesia in anesthetized mice.The Morris water maze test, open field test and fear conditioning test were performed at 1 day before operation and at 1, 3 and 7 days after operation.At 1, 3 and 7 days after operation, 3 mice with worst cognitive performance in each cognitive function assessments were sacrificed in group P, and three mice were randomly sacrificed in group C. The hippocampal tissues were then obtained, the expression of differentially expressed proteins was identified by high-performance liquid chromatography-mass spectrometry, and Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to analyze the differentially expressed proteins. Results:Compared with group C, the escape latency at different time points was significantly prolonged, and the percentage of time spend on target quadrant and the percentage of freezing time in fear conditioning test were decreased in group P ( P<0.05). There were 21 differentially expressed proteins, of which 12 proteins showed up-regulated expression and 9 proteins showed down-regulated expression.The GO functional analysis showed that the differentially expressed proteins were involved in the process such as the metabolism, signal transmission, regulation of biological processes, formed cell components such as synapses and organelles, and were related to molecular function such as binding and transportation.KEGG signaling pathway analysis showed that there were also differences in MAPK signaling pathway, ErbB signaling pathway, AMPK signaling pathway and the transport of SNARE protein in vesicle and etc. Conclusion:There are 21 differentially expressed proteins in the hippocampus of PND mice, and these proteins are involved in the pathophysiological process probably related to PND such as neuroinflammatory responses, abnormal synaptic structure, mitochondrial dysfunction and decreased autophagy.

Objective:To explore the effect of group cognitive behavior therapy (GCBT) on anxiety, depression and quality of life in patients with chronic obstructive pulmonary disease (COPD) in community.Methods:From August to November 2019, patients with moderate COPD in 18 communities in Xuzhou City were randomly divided into the intervention group ( n=240) and the control group ( n=223). The control group received routine management and the intervention group received group cognitive behavioral therapy intervention for 8 weeks on the basis of routine management.Before and after the intervention, FEV 1% predicted value and FEV 1/FVC were measured by pulmonary function tester.Hospital Anxiety and Depression Scale (HADS) was used to evaluate the anxiety and depression of patients.St.George's respiratory questionnaire (SGRQ), COPD assessment test (CAT) and modified medical research council dyspnea (mMRC) were used to evaluate the quality of life of patients.SPSS 20.0 software was used for analysis.The χ 2 test, independent sample t-test, paired sample t-test were used for statistical analysis. Results:After 8 weeks of intervention, the anxiety and depression scores of the intervention group were lower than those of the control group (anxiety: (8.23±4.02) vs (10.71±3.60); depression: (7.87±3.73) vs (10.20±3.72)( t=6.415, 6.185, both P<0.01). After the intervention, there was no significant difference in FEV 1%((51.7±12.3)% vs (52.0±12.6)%) predicted value and FEV 1/FVC((57.3±10.8)% vs (56.9±10.7)%) between the two groups( t=-0.259, 0.400, both P>0.05). The scores of CAT, mMRC and SGRQ in the intervention group were lower than those in the control group((17.35±5.78) vs (20.90±8.00), (1.55±0.82) vs (2.30±1.21), (41.78±21.56) vs (57.08±24.46))( t=-5.061, -7.227, -6.580, all P<0.01). Conclusion:Group cognitive behavioral therapy can relieve the anxiety and depression and improve the quality of life of patients with COPD.

Objective:To evaluate the association between preoperative serum β 2-microglobulin (β 2MG) concentrations and postoperative delirium (POD) in elderly patients. Methods:The study selected patients who underwent knee or hip arthroplasty under spinal-epidural anesthesia on an elective basis at Qingdao Municipal Hospital from May 2021 to November 2022. The patients were divided into a POD group and a non-POD group based on the occurrence of POD. The study was conducted as part of the Perioperative Neurocognitive Impairment and Biomarkers Lifestyle Cohort, which was a nested case-control study. The study collected baseline data from two groups of patients and analyzed the differences between them. Logistic regression was used to identify the risk factors for POD. The stability of the regression model was tested using sensitivity analysis. The mediation model was used to examine whether cerebrospinal fluid (CSF) biomarkers mediated the relationship between β 2MG and POD. The receiver operating characteristic curve was drawn and the area under the curve was calculated to evaluate the accuracy of preoperative β 2MG concentrations and CSF biomarker concentration in predicting POD. Results:There were 57 cases in POD group and 449 cases in non-POD group. The results of logistic regression analysis showed that the increased β 2MG and CSF total tau protein (t-tau) concentrations were risk factors for POD, and the increased CSF β-amyloid 42 concentration was a protective factor for POD after adjustment for multiple confounders such as age, gender, education, Mini-Mental State Examination, history of hypertension and infusion volume ( P<0.05). The results of mediation analysis showed that the serum β 2MG′s effect on POD was partly mediated by t-tau (18.1%). The results of the receiver operating characteristic curve showed that the area under the curve of the β 2MG concentration combined with the CSF biomarker concentration was 0.742. Conclusions:Elevated preoperative serum β 2MG concentration is a risk factor for POD in elderly patients, and the relationship may be partly mediated by CSF t-tau.

Objective:To evaluate the relationship between preoperative serum bilirubin concentrations and postoperative delirium (POD) in the patients undergoing knee or hip replacement.Methods:Medical records from 413 patients undergoing knee or hip arthroplasty were selected from August 2020 to October 2023 at Qingdao Municipal Hospital using a nested case-control design based on the PNDABLE study cohort. The patients were divided into POD group ( n=77) and non-POD group ( n=336) according to whether POD occurred. Univariate analysis was used to analyze the influencing factors. Logistic regression was used to identify the risk factors for POD. The significance of mediation effect was tested. The receiver operating characteristic curve was drawn to evaluate the accuracy of risk factors in predicting POD. Results:There were significant differences in age, education time, ratio of diabetes history, Memorial Delirium Assessment Scale score, total bilirubin concentration, direct bilirubin concentration, indirect bilirubin concentration, Aβ 42 concentration, p-tau concentration, t-tau concentration, Aβ 42/p-tau ratio and Aβ 42/t-tau ratio between POD group and non-POD group ( P<0.05). The results of logistic regression analysis showed that preoperative serum total bilirubin, direct bilirubin and indirect bilirubin were risk factors for POD ( P<0.05). The results of mediation effects showed that the concentration of total tau protein in CSF partly mediated the relationship between high serum total bilirubin, direct bilirubin and indirect bilirubin concentrations and POD. The area under the receiver operating characteristic curve of total bilirubin, direct bilirubin and indirect bilirubin combined with CSF biomarker concentrations in predicting POD was 0.83 ( P<0.001). Conclusions:Preoperative elevated concentrations of total bilirubin, direct bilirubin and indirect bilirubin are risk factors for POD in the patients undergoing knee or hip replacement. CSF t-tau concentration has a partly mediating role in the association between serum total bilirubin, direct bilirubin and indirect bilirubin concentrations and the development of POD.

Objective To explore the relationship between different types of female reproductive system dysplasia and age of visit, clinical manifestations, common types of combined malformations and endometriosis. Methods The patient′s medical records in the Second Hospital of Hebei Medical University from December 2002 to June 2016 were collected and retrospectively analyzed. Results Among 924 cases of genital tract dysplasia, uterine dysplasia (65.3%, 824/1 261) was the most common, followed by vaginal dysplasia (28.3%, 357/1 261), hymen atresia and urogenital fistula (3.7%, 47/1 261), and cervical dysplasia (2.6%, 33/1 261). (1) The youngest age was in patients with hymen atresia and urogenital fistula, with a median of 14.5 years old, while the older age were in patients with uterine, vaginal and cervical dysplasia, with median age of 25.0, 24.0 and 23.0 years old, respectively. (2) The clinical manifestations were lack of specificity, mainly abnormal findings of physical examination or accessory examination, primary amenorrhea, lower abdominal pain, infertility, adverse pregnancy history. (3) About other systemic malformations, urological malformations were the most common (4.8%, 44/924), followed by spinal malformations (0.5%, 5/924), inguinal hernia (0.4%, 4/924), heart malformations (0.2%, 2/924), cleft lip and palate (0.2%, 2/924). Oblique vaginal septal syndrome and MRKH syndrome were the most likely to be associated with other system malformations. (4) About combination with endometriosis, there was no significant difference between obstructive genital tract malformations (2.3%, 9/385) and non obstructive genital tract malformations (1.7%, 9/539; P=0.469). Conclusions Female reproductive system dysplasia is the most common in uterine dysplasia, followed by vaginal dysplasia, hymen atresia and urogenital fistula, and cervical dysplasia. The age of visit is generally older, often found by abnormal findings of physical examination or accessory examination, primary amenorrhea, lower abdominal pain, infertility, adverse pregnancy history;and could be combined with a variety of other system malformations, most seen by urinary system malformations,there is also the risk of endometriosis.

Objective:To evaluate the relationship between hippocampal macrophage polarization and perioperative neurocognitive disorders in mice with tibial fractures.Methods:Forty-five clean-grade healthy male C57/BL6 mice, aged 5-7 months, were divided into 3 groups ( n=15 each) using the random number table method: control group (group C), anesthesia group (group A) and anesthesia surgery group (group AS). Group C received no treatment. Group A was anesthetized with isoflurane inhaled for 15 min. In AS group, intramedullary nail fixation of tibial fracture was performed under anesthesia through inhalation of 2% isoflurane. Morris water maze test and open field test were performed before anesthesia/on 1 day before surgery and after anesthesia/on 1, 3 and 7 days after operation. Five mice were randomly selected after the behavioral experiments were completed at each time point, and hippocampal tissues were taken after the animals were sacrificed for determination of the expression of tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), IL-6, chemokine (c-c motif) ligand 12 (CCL2), CCL5, CCL8, iNOS and Arg-1 mRNA (by quantitative real-time polymerase chain reaction), expression of iNOS and Arg-1 proteins (by Western blot), and percentage of CD11b, CD45, CD86 and CD206 cells in hippcampal area (by immunofluorescence staining). Results:Compared with group C, the escape latency was significantly prolonged after operation, the number of crossing the platform was reduced after operation, the expression of TNF-ɑ, IL-6, CCL5 and CCL8 mRNA and iNOS protein and mRNA was up-regulated, the expression of Arg-1 protein and mRNA was down-regulated, the percentages of CD11b + CD45 + cells and CD11b + CD86 + cells in the hippocampus were increased, and the percentages of CD11b + CD206 + cells were decreased in AS group ( P<0.05), and no significant change was found in the parameters mentioned above in group A ( P>0.05). Compared with group A, the escape latency was significantly prolonged after surgery, the frequency of crossing the platform was reduced after surgery, the expression of iNOS mRNA was up-regulated, the percentages of CD11b + CD45 + cells and CD11b + CD86 + cells were increased, and the percentages of CD11b + CD206 + cells were decreased in AS group ( P<0.05). Conclusions:The occurrence of PND may be related to increased polarization to M1 macrophages in the hippocampus and decreased polarization to M2 macrophages in mice with tibial fracture, which further leads to central inflammatory responses.