BackgroundChronic insomnia is characterized by a prolonged and recurrent course. The efficacy of repeated transcranial magnetic stimulation （rTMS） as a physical therapy method to improve sleep quality remains inadequately supported by evidence， particularly regarding its relationship with personality traits. ObjectiveTo explore the efficacy and influencing factors of rTMS in the treatment of chronic insomnia， and to provide insights into its therapeutic potential. MethodA total of 46 patients who met the diagnostic criteria for chronic insomnia according to the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5）， and were treated at the Third Hospital of Mianyang from September 2022 to September 2023 were selected. Prior to treatment， participants underwent assessments using the Eysenck Personality Questionnaire （EPQ）， Hamilton Depression Scale-17 item （HAMD-17） and Hamilton Anxiety Scale （HAMA）. The Pittsburgh Sleep Quality Index （PSQI） was used to assess sleep quality before treatment， at the end of the second week of treatment and one week post-treatment. ResultsAt the end of the second week of treatment， patients exhibited significantly improved total PSQI score and subscale scores related to subjective sleep quality， sleep latency， sleep duration， sleep efficiency， sleep disturbance and daytime dysfunction （t=4.755~13.361， P<0.01）， with 24 cases （54.35%） showing effective treatment outcomes. Multiple linear regression analysis showed that introverted and extroverted personality traits contributed significantly to the regression equation （B=0.317， P<0.01）， explaining 29.90% of the total variation （R²=0.299）. ConclusionrTMS treatment may effectively improve the sleep quality of patients with chronic insomnia， with its therapeutic effect appearing to associated with introverted and extroverted personality traits. ［Funded by National Natural Science Project of China （number， 82372080）］

ObjectiveTo explore the effect of precocious puberty on glucose metabolism and lipid metabolism in female rats. MethodsSixty two-day-old female rats were randomly divided into 2 groups. When aged 5 days， the precocious puberty group and normal group were given a single subcutaneous injection of danazol and solvent soybean oil respectively. The vaginal opening of rats was monitored from their 21 days of age. After 12 hours of fasting， all successful modeling rats were randomly executed within 3 days after vaginal opening， when aged 7 and 12 weeks. Then we measured the rats’ body weight and length， determined the concentrations of glucose， insulin， blood lipids， estradiol， leptin and adiponectin with enzyme-linked immunosorbent assay and observed the pathological changes of perirenal fat， uterus and ovary. ResultsFor body weight and length， rats in the precocious puberty group were smaller than those in the normal group within 3 days after vaginal opening， but which did not affect their subsequent growth and development， and there was no significant difference between the two groups at 7 and 12 weeks of age. Within 3 days after vaginal opening， insulin levels had significant difference between the two groups （P = 0.001）， the precocious group showed hyperinsulinemia and increased number of perirenal adipocytes. At three execution times， no significant difference was noted in estradiol， leptin and adiponectin levels between the two groups. The same was true in the ratios of ovary or uterus to body weight between the two groups. ConclusionsPrecocious puberty makes earlier onset of pubertal development and allows body maladaptation to the sudden changes of the internal environment. However， the changes due to precocious puberty are temporary and reversible， and they may become normal in adulthood.

Objective To investigate the feasibility of granzyme B(GB)promoter-controlled ferritin heavy chain(FTH1)reporter gene expression for monitoring the activation status of chimeric antigen receptor T cells(CAR-T)by magnetic resonance imaging(MRI).Methods Cytotoxic T lymphocytes(CTLs)were screened by Ficoll density gradient centrifugation and flow sorting.The GB promoter and FTH1 gene were ligated together with disialoganglioside 2(GD2)CAR,and lentiviral vectors were transfected into CTLs to construct GD2-CAR-T/pGB-FTH1 cells.GD2-CAR-T/pCMV-FTH1,GD2-CAR-T,and T cells served as control cells.CytoTox96@non-radioactive cytotoxicity was used to detect the killing effect of each group of cells after co-culture with human neuroblastoma cells(SK-N-SH).ELISA was employed to detect the coincubation factor as well as the amount of GB secretion.Western blotting,Prussian blue staining and cellular MRI were applied to detect the expression of the FTH1 gene after co-culture.Results CTLs were successfully obtained,and then GD2-CAR-T/pGB-FTH1,GD2-CAR-T/pCMV-FTH1 and GD2-CAR-T cells were constructed.The killing effect,co-incubation factor and GB secretion of the above 3 groups of cells were significantly higher than those of the T cells,and the level of GB expression was highest at day 1,and then decreased in order at day 3 and day 7 after co-culturing with SK-N-SH cells.The relative expression of FTH1 and iron content of the GD2-CAR-T/pGB-FTH1 cells showed the same trend as GB expression,and the MRI signals were gradually increased.There were no significant differences in the relative expression of FTH1,iron content and MRI signals in the GD2-CAR-T/pCMV-FTH1 cells at all time points.No FTH1 expression or iron aggregation was observed in the GD2-CAR-T and T cells groups.Conclusion MRI based on the FTH1 reporter gene driven by the granzyme B promoter can reflect the GB expression level and tumor killing effect of CAR-T cells,which provides a potential real-time visual means to monitor the cell activation status for CAR-T therapy.

Objective To establish and optimize a method for simultaneous determination for the contents of sodium,potassium,calci-um and magnesium ions in serum using microwave digestion combined with ion chromatography,and evaluate its performance.Methods Serum samples were pretreated by microwave digestion,and four cations were quantitatively determined using ion chromatography.The linearity,precision,recovery and accuracy of the established method were verified with reference to the EP6-A and EP15-A3 docu-ments issued by the Clinical and Laboratory Standards Institute(CLSI).Results The imprecisions of determinations for sodium,po-tassium,calcium and magnesium ions expressed as coefficient of variation(CV)were 0.10％to 0.36％,0.15％to 0.48％,0.22％to 0.87％and 0.21％to 0.73％,respectively.The recoveries with adding standard of the four cations were between 97.3％and 103.0％.Referred to RELA2020 and RELA2021,the deviations of the analysis results were-0.57％to-0.14％,-0.84％to-0.16％,-1.82％to-0.37％,and-0.60％to 0.34％for sodium,potassium,calcium,and magnesium,respectively,and all the comparisons were pas-sed.The limits of detection/quantification were 0.017 5/0.058 3 mmol/L,0.000 5/0.001 7 mmol/L,0.009 9/0.032 9 mmol/L and 0.001 1/0.003 5 mmol/L for the four cations,indicating that after pretreatment,no significantly interfere from other impurities for the results of determination was found.Conclusion A method for simultaneous determination of sodium,potassium,calcium and magnesi-um ions in serum was successfully established and optimized using microwave digestion combined with ion chromatography.The method showed good precision and accuracy,simple operation,short time consumption and low cost,thus it could be used for quality improve-ment of clinical electrolyte testing in serum.

Objective:To develop a candidate reference method for cortisol in human serum by isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC/MS/MS).Methods:An isotope-labeled internal standard was added to samples, followed by alkalizing with sodium carbonate solution and liquid-liquid extraction with n-hexane/ethyl acetate. Isoelution with a methanol aqueous solution was employed for liquid chromatographic separation, while ESI in the positive ion and multiple reaction monitoring mode were used for mass spectrometry. The volume of sample, standard, and internal standard solutions were all controlled by weighing, and the results were calculated by bracketing method. The accuracy, precision, specificity, linearity, LOD and LOQ of this method were evaluated referring to the CLSI C62-A and C50-A guidelines and the domestic expert consensus documents.Results:The method demonstrated excellent accuracy and precision with the bias of ERM-DA192, ERM-DA193 and RELA 2021-2023 all being less than 2%. The recovery of added cortisol ranged from 98.2% to 101.1%. Both intra- and inter-assay imprecisions was <2%. The method was free from interference by structural analogue and showed a good linearity in the range of 25-1 600 nmol/L. The LOD and LOQ were 0.5 nmol/L and 1.0 nmol/L, respectively. A cortisol assay kit (chemiluminescence immunoassay) traced to this candidate reference method was used to determine 46 clinical serum samples concurrently, and the two methods exhibited good correlation.Conclusions:A candidate reference method for the determination of cortisol in serum was established, demonstrating high sensitivity, good repeatability and accuracy. This method can serve as a reference for the measurement traceability and accuracy evaluation of routine methods.

Objective:To compare the application value of Caprini thrombosis risk assessment scale and Shanghai venous thrombosis risk factor scale in predicting venous thromboembolism in hospitalized maternal patients.Methods:This was a case-control study. A total of 67 pregnant women complicated with venous thromboembolism in the Obstetrics Department of Shandong Provincial Hospital Affiliated to Shandong First Medical University from January 2010 to September 2021 were retrospectively analyzed. And 144 pregnant women without venous thromboembolism in the same period were selected. Two venous thrombosis risk assessment tools were used to evaluate the pregnant women, and the predictive effectiveness and accuracy of the two assessment tools were compared.Results:The effects of the two risk assessment tools for venous thrombosis were different before and after delivery with statistical difference ( Z=8.15, 5.97, both P<0.01), but the Shanghai venous thrombosis risk factor scale (83.9%, 67.3%) was superior to Caprini thrombosis risk assessment scale (52.1%, 45.0%) in the accuracy of prenatal and postnatal prediction. The area under receiver operating characteristic curve showed that the Shanghai venous thrombosis risk factor rating scale (0.863) was significantly superior to the Caprini thrombosis risk assessment scale (0.748) after delivery. Conclusions:The Shanghai venous thrombosis risk factor scale is more valuable than the Caprini thrombosis risk assessment scale in the early risk identification of venous thromboembolism in pregnant women.

OBJECTIVE To provide a reference for the safe use of drugs in patients with complex venous thromboembolism （VTE） and acute renal insufficiency. METHODS Clinical pharmacists participated in the management of anticoagulant therapy for a patient with complex VTE complicated with acute renal insufficiency， and evaluated the patient as high-risk thrombosis and bleeding based on their medical history， laboratory test results， etc.； combined with the complexity of thrombosis and renal insufficiency， clinical pharmacists suggested that enoxaparin sodium should be used in the acute stage of thrombosis （5 to 21 days after onset）， and then warfarin should be adopted for oral anticoagulation treatment. Because the patient’s anticoagulation was not up to the standard （the target range of the international normalized ratio was 2-3）， clinical pharmacists suggested increasing the warfarin dose， detecting the warfarin metabolism genotype， and adjusting the warfarin dose according to the genotype； at the same time， clinical pharmacists developed an anticoagulation monitoring plan to ensure the safety of anticoagulation treatment. RESULTS Doctors had adopted all the recommendations of clinical pharmacists. The patient did not experience adverse events such as bleeding or worsening of thromboembolism during anticoagulation in the hospital. When the anticoagulation met the standards， the patient was allowed to be discharged with medication. CONCLUSIONS By participating in the anticoagulation treatment management of patients with complex VTE and acute renal insufficiency， clinical pharmacists have assisted doctors in formulating personalized anticoagulation plans to promote the compliance with the anticoagulation treatment standard and ensure the safety and effectiveness of medication for patients.

Objective To analyze the educational characteristics of physical therapy in universities of United States, and provide reference for the international development of rehabilitation therapy education in China.Methods Based on the data collected from the official websites of three universities in the United States and the databases at home and abroad, combined with the author's learning experience, the characteristics of physical therapy education in universities of United States in detail was analyzed.Results Characteristics of physical therapy education in universities of United States had strict accreditation system, course objectives oriented by practicing competence, comprehensive curriculum plan, educational concept of student-centered, high level of teaching informatization, diversified teaching and learning evaluation system, which conformed to the goal of rehabilitation competency of World Health Organization.Conclusion Physical therapy education in United States has remarkable characteristics and a well-developed system, which has certain reference value for China.

Objective To evaluate therapeutic effects of bone marrow mesenchymal stem cells(MSC)derived exosomes on alcohol-induced liver injury.Methods Eighteen male C57BL/6 mice aged 6 to 8 week were randomly divided into control group,model group and exosomes group,with 6 mice in each group.The mice in the model group and the exosomes group were fed with Lieber-DeCarli ad libitum diet(Dyets Inc.)for 4 weeks,followed by gavage a bolus of ethanol at day 26,27 and 28.The mice in the control group matched the alcohol-derived calories with dextran-maltose.Meanwhile,the mice in exosomes group were injected with MSC-exosomes via the tail vein at day 14 and 26.After the experiment,serum levels of alanine aminotransferase(ALT)and aspartate aminotransaminase(AST)were detected,and the pathological changes of liver tissues were observed.The expressions of nuclear factor erythroid 2-related factor 2(Nrf-2),heme oxygenase-1(HO-1),CD63,CD81,TSG101 and Cytochrome C were analyzed by Western blot,and mRNA levels of Nrf-2,HO-1,interleukin(IL)-10 and IL-17 were analyzed by real-time polymerase chain reaction(RT-PCR).The commercial kits were used to detect serum IL-10,IL-17 levels and liver tissue malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD)oxidative stress indicators.The numbers of regulatory T cell(Treg)and help T(Th)17 cells in the liver were analyzed by flow cytometry.One-way analysis of variance was used for comparison between groups.Results MSC-exosomes expressed positive markers CD63,CD81 and TSG101,but did not express the negative markers Cytochrome C.The serum ALT and AST levels in model group were(87.3±25.1)U/L and(223.2±43.5)U/L,respectively,while those in exosomes group were(47.7±12.0)U/L and(128.2±33.6)U/L,respectively.The differences between the two groups were both statistically significant(F=12.818 and 12.226,respectively,both P<0.05).Compared with control group,the SOD activity and GSH level in the model group significantly decreased with statistically significant differences(F=4.245 and 24.074,respectively,both P <0.05).Lieber-DeCarli ethanol feeding significantly increased intrahepatic MDA level in the model mice,which was reversed by MSC-exosomes supplementation,and the difference was statistically significant(F=36.675,P <0.05).Compared with control group,the intrahepatic protein expressions of Nrf-2 and HO-1 in model group were significantly decreased,while the expressions in exosomes group were obviously increased.The differences were statistically significant(F=33.623 and 14.960,respectively,both P <0.05).The expression trends of Nrf-2 and HO-1 mRNA were the same as those of protein expressions(F=20.784 and 276.336,respectively,both P <0.05).The proportions of liver Treg/Th17 in the control group,model group and exosomes group were 4.3±0.9,0.4±0.2,and 3.4±0.5,respectively.The differences among groups were statistically significant(F=64.227,P <0.05).Compared with control group,the serum protein and intrahepatic gene expression of IL-17 in the model group were significantly increased,which were reversed by MSC-exosomes treatment.The differences were statistically significant(F=15.581 and 40.095,respectively,both P<0.05).Serum IL-10 protein levels and intrahepatic IL-10 gene expression were significantly decreased after Lieber-DeCarli ethanol feeding,which were lower than the exosomes group.The differences were statistically significant(F=98.268 and 153.743,respectively,both P <0.05).Conclusions MSC-exosomes transplantation may relieve alcohol-induced liver injury.The mechanism could involve reduction of oxidative stress in the liver via regulating Nrf-2/HO-1 and normalizing the balance of Treg and Th17 cells.

Objective To examine the effect of adolescent intermittent ethanol exposure (AIE) on ethanol-induced conditioned place preference (CPP) and anxiety-like behavior in adolescent mice. Methods In experiment 1, adolescent male C57BL/6 mice at 4 weeks of age were randomly divided into AIE group and NS group (n=10 for each group). The binge drinking model was established by AIE (3 g/kg, 25%). The alcohol reward was evaluated using the ethanol-induced CPP paradigm (2 g/kg, 20%). In experiment 2, the anxiety-like behavior of adolescent male C57BL/6 mice were assessed using the elevated plus maze (EPM) test, and the animals were then allocated into high-anxiety mouse (HAM) and low-anxiety mouse (LAM) groups based on the percentage of open arm time (OT%). HAM and LAM were randomly divided into AIE group and NS group (n=8～10 for each group) with random number method, respectively. Then, anxiety-like behavior in four groups was measured again using the EPM test. Results In experiment 1. Ethanol preference (116.1± 12.9)s vs. (70.8±14.8)s, P=0.035) was significantly higher in AIE group relative to NS group. However, In experiment 2. The alteration in anxiety-like behaviors was not significant in either HAM-AIE or LAM-AIE groups (all P＞0.05). Conclusions AIE reinforces ethanol-induced CPP but does not affect the anxiety-like behavior in adolescent mice, suggesting that AIE may not play a role in anxiety-like behavior.