Objecave To develop a sensitive and specific LC-MS/MS method for determination of testolactone in human urine.Methods A C_(18 )column(2.1×50mm,3.5μm) was used.The mobile phase Was a mixture of acetonitrile and the buffer solution(ammonium acetate-water solution adjusted with formic acid to pH 3.5)at a flow rate of 0.5ml/min.A mass spectrometer equipped with electrospray ionization source was used as a detector and operated in the positive mode.In multiple reaction monitoring(MRM)mode,the ion transitions of m/z 301→121 and m/z 301→25 was used to qualify and quantify the testolactone,respectively.Results Chromatograms showed no endogenous interfering peaks with the urine blank sample.Each analysis was completed within 7min The calibration wag linear in the concentration range within 0.1～50μg/ml.The intra-batch and inter-batch RSD were less than 10%.The recovery rate of the extraction was about 60%.Conclusions The method is proved to meet the requirements of WADA and be suitable for routine screening.

Objective To evaluate the changes in motor function impairment and brain functional networks of patients with acute ischemic stroke(AIS)through parameters of spontaneous activities of both upper limbs and electroencephalogram graph theory analysis methods.Methods The data of 34 acute ischemic stroke patients(observation group)with upper limb motor disorders who were treated in the Department of Neurology of the Affiliated Hospital of Southwest Medical University from January 2022 to October 2023,and 40 healthy controls(HC group)were collected.The subjects completed the National Institutes of Health Stroke Scale(NIHSS)and Fugl-Meyer Assessment(FMA)within 7 days,and wore wrist activity recorders(Actiwatch)continuously for 24 hours to collect data on sponta-neous activities of upper limbs and analyzed related parameters such as the coordination coefficient of both upper limbs(r),the activity ratio of the affected side to the healthy side upper limb(ULAR),etc.At the same time,all subjects completed approximately 2 hours of 19-channel electroencephalogram examination.After preprocessing the electroencephalogram data,5 segments of 10-second resting-state electroencephalogram were extracted for graph theory.Results ① Compared to healthy individuals,AIS patients exhibited decreased functional connectivity edges in the 8 and θ bands,with substantial reductions in network connections in the α band.In the β band,connections between the frontal,right parietal,and occipital regions weakened,while connections from the right temporal lobe to the left temporal lobe strengthened.In the γ band,there was a significant increase in connections throughout the brain.② Graph theory analysis revealed significantly increased shortest path lengths(α band:t=2.228,P＜0.05,d=-0.52;β band:t=-3.641,P＜0.01,d=-0.878)and decreased global efficiency(α band:t=2.535,P＜0.05,d=0.591;β band:t=3.321,P＜0.01,d=0.803)in the observation group compared to the control group.In the γ band,local effi-ciency(t=3.279,P＜0.01,d=0.765)and clustering coefficients were significantly higher(t=3.358,P＜0.01,d=0.783).③ In the-γ band,the ULAR ≤30％group showed significantly re-duced shortest path length(t=-2.063,P＜0.05,d=-0.802)and increased global efficiency(t=2.226,P＜0.05,d=0.865),local efficiency(t=2.95,P＜0.05,d=1.147),and cluste-ring coefficient(t=2.962,P＜0.05,d=1.148).④ In the observation group,the bilateral upper limb coordination coefficient during sleep was negatively correlated with NIHSS scores(r=-0.389,P＜0.05)and ULAR(r=-0.395,P＜0.05),while FMA scores were positively cor-related with ULAR(r=0.442,P＜0.05).Conclusion The parameters of spontaneous activities of the upper limbs can be used to determine the impairment of motor function in AIS patients.The combination of changes in brain functional networks and motor impairments can provide new ideas for the study of their neural network mechanisms.

Objective To evaluate the changes in motor function impairment and brain functional networks of patients with acute ischemic stroke(AIS)through parameters of spontaneous activities of both upper limbs and electroencephalogram graph theory analysis methods.Methods The data of 34 acute ischemic stroke patients(observation group)with upper limb motor disorders who were treated in the Department of Neurology of the Affiliated Hospital of Southwest Medical University from January 2022 to October 2023,and 40 healthy controls(HC group)were collected.The subjects completed the National Institutes of Health Stroke Scale(NIHSS)and Fugl-Meyer Assessment(FMA)within 7 days,and wore wrist activity recorders(Actiwatch)continuously for 24 hours to collect data on sponta-neous activities of upper limbs and analyzed related parameters such as the coordination coefficient of both upper limbs(r),the activity ratio of the affected side to the healthy side upper limb(ULAR),etc.At the same time,all subjects completed approximately 2 hours of 19-channel electroencephalogram examination.After preprocessing the electroencephalogram data,5 segments of 10-second resting-state electroencephalogram were extracted for graph theory.Results ① Compared to healthy individuals,AIS patients exhibited decreased functional connectivity edges in the 8 and θ bands,with substantial reductions in network connections in the α band.In the β band,connections between the frontal,right parietal,and occipital regions weakened,while connections from the right temporal lobe to the left temporal lobe strengthened.In the γ band,there was a significant increase in connections throughout the brain.② Graph theory analysis revealed significantly increased shortest path lengths(α band:t=2.228,P＜0.05,d=-0.52;β band:t=-3.641,P＜0.01,d=-0.878)and decreased global efficiency(α band:t=2.535,P＜0.05,d=0.591;β band:t=3.321,P＜0.01,d=0.803)in the observation group compared to the control group.In the γ band,local effi-ciency(t=3.279,P＜0.01,d=0.765)and clustering coefficients were significantly higher(t=3.358,P＜0.01,d=0.783).③ In the-γ band,the ULAR ≤30％group showed significantly re-duced shortest path length(t=-2.063,P＜0.05,d=-0.802)and increased global efficiency(t=2.226,P＜0.05,d=0.865),local efficiency(t=2.95,P＜0.05,d=1.147),and cluste-ring coefficient(t=2.962,P＜0.05,d=1.148).④ In the observation group,the bilateral upper limb coordination coefficient during sleep was negatively correlated with NIHSS scores(r=-0.389,P＜0.05)and ULAR(r=-0.395,P＜0.05),while FMA scores were positively cor-related with ULAR(r=0.442,P＜0.05).Conclusion The parameters of spontaneous activities of the upper limbs can be used to determine the impairment of motor function in AIS patients.The combination of changes in brain functional networks and motor impairments can provide new ideas for the study of their neural network mechanisms.

Objective To investigate the relationship between cellular immune response which is related with Th1/Th2 cells and the different severities of tuberculosis pleural effusion adhesion.Methods A total of 66 in-patients diagnosed with different severities of tuberculosis pleural effusion adhesion by internal thoracoscope were enrolled from August 2014 to December 2016.ELISA was used to determine levels of INF-γ,TNF-α,IL-2 and IL-4. The ratio of Th1 and Th2 cells was detected by flow cytometry. Results All the patients were divided into 3 groups:9 cases without pleural adhesion,32 cases with mild pleural adhesion,25 cases with severe pleural adhe-sion. The levels of 4 cytokines in pleural fluid were significantly higher than those in serum in each group(P <0.05,respectively).The concentrations of INF-γ and TNF-α were increased with the severity of tuberculosis pleu-ral effusion adhesion. The levels of INF-γ and TNF-α in the severe pleural adhesion group were markedly higher than those in the other two groups(P<0.05,respectively).The proportion of Th1 cells in the severe pleural adhe-sion group was significantly higher than that in the none pleural adhesion group and in the mild pleural adhesion group(P<0.05,respectively).Conclusions The proportion of Th1 cells and levels of INF-γ and TNF-α are pos-itively related with pleural adhesion severity. Cellular immune response which is related with Th1 cells contributes to pathological immune pleural damage and intensify the severity of pleural adhesions.

Objective To compare the concentrations of alpha-enolase (ENO1),CYFRA21-1,and CA125 in the patients with malignant pleural effusion ,tuberculous exudative pleural effusion ,or parapneumonia pleural effusion. To explore the clinical value of ENO1 in pleural effusion combined with serum CYFRA21-1 and CA125 in diagnosis of malignant pleural effusion. Methods Enzyme-linked immunosorbent assay(ELISA)was used to detect the concentration of ENO1 in pleural effusions. The concentrations of CA125 and CYFRA21-1 in the blood samples were measured using chemiluminescence and magnetic particle-based chemiluminescence respective-ly. The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection were calculated. Results The concentration of ENO1 in malignant pleural effusion group was significantly increased(P<0.001);the concentrations of ENO1 did not differ significantly between tuberculous exudative pleural effusion and parapneu-monia pleural effusion(P>0.05). The sensitivity and specificity of ENO1 combined with serum CYFRA21-1 and CA125 detection in malignant pleural effusion were 94％ and 74％,98％ and 98％,respectively. Conclusions ENO1 combined with serum CYFRA21-1 and CA125 detection can improve the sensitivity of diagnosis of malignant pleural effusion and enhance the diagnostic rate of malignant pleural effusion.