Objective To establish the preliminary reference intervals for the percentage and fluorescence intensity ratio(FIR)for 12 platelet-leukocyte aggregates(PLAs)in peripheral blood circulation.Methods A total of 124 healthy individuals(61 males and 63 females)aged 18-90 years were selected from Beijing Jishuitan Hospital.Venous blood samples were collected in EDTA-K2 tubes for anticoagulation,and flow cytometry was used to measure the percentage and FIR of 12 types of PLAs.All the participants were grouped by gender for comparative analysis.Reference intervals for each parameter were calculated according to the WS/T 402-2024 standard.Results The percentages of platelet-T lymphocyte aggregates,platelet-CD4+T lymphocyte aggregates,and platelet-CD8+T lymphocyte aggregates in males were significantly lower than those in females(all P＜0.05).The FIRs of platelet-T lymphocyte aggregates,platelet CD8+T lymphocyte aggregates,platelet-B lymphocyte aggregates,and platelet-NK cell aggregates in males were significantly higher than those in females(all P＜0.05).Conclusion The preliminary reference intervals for the percentage and FIR of 12 types of PLAs in healthy adults have been established.Different gender may influence the detection results of platelet-leukocyte aggregates,especially platelet-lymphocyte aggregates,and the corresponding FIR values in healthy adults.Further large-scale studies should be necessary to confirm these findings.

Objective:To analyze the phenotypes and gene mutations of the three families with hereditary protein S (PS) deficiency caused by frameshift heterozygous variants.Methods:Case report and case series study. We investigate three probands and their family members (14 people from three generations) who registrated in Beijing Jishuitan Hospital of Capital Medical University from Feb 1st to 28th, 2025. The clinical data of the three probands and their family members were collected. The related coagulation tests of all members were detected. Protein S activity (PS:A) was determined using coagulation assay, and total protein S antigen (TPS:Ag) and free protein S antigen (FPS:Ag) were measured using ELISA. All exons and their flanking sequences of the probands were amplified using PCR technique, and gene analysis by direct sequencing, and variant genes were searched which were then verified by cloning sequencing, meanwhile, the corresponding variant loci of the family members were analyzed. A calibrated automated thrombin generation (CAT) method was used to study thrombin production. ClustalX-2.1-win software was used to analyze the conservatism of the mutant loci; Mutation Taster and Franklin.genoox online bioinformatics software were used to predict the pathogenicity of the mutant loci; PyMol software was used to analyze the changes in protein spatial structure before and after mutation.Results:The PS:A, TPS:Ag and FPS:Ag of the three probands with hereditary PS deficiency were decreased. Genetic sequencing identified a total of three PROS1 genetic variants, and all the three probands were heterozygous frameshift variants: p.Gln51Argfs*2 in Proband A; p.Met251Valfs*17 in Proband B; and Thr478tyrfs*21 in Proband C. Conservative analysis showed that p.Gln51, p.Met251 and p.Thr478 loci were not conserved among the homologous species; Mutation Taster and Franklin.genoox analysis showed that these variants were pathogenic; protein structure model analysis showed that p.Gln51Argfs*2, p.Met251Valfs*17 and p. Thr478Tyrfs*21 variants may result in altered protein spatial structure.Conclusion:The heterozygous frameshift variations of PS p.Gln51Arg fs*2, p.Met251Valfs*17 and p.Thr478Tyrfs*21 would alter the spatial structure of PS molecules, and reduce their structural stability, leading to PS deficiency.

Objective To analyze the current status and influencing factors of occupational stress, job burnout and sleep quality among workers in the secondary industry in Jinshan District, Shanghai City. Methods A total of 1 418 workers from six key industries in Jinshan District, Shanghai City were selected as the study subjects by the stratified cluster sampling method. The Occupational Stress Core Scale, Maslash Burnout Inventory General Survey and Pittsburgh Sleep Quality Index were used to investigate occupational stress, job burnout and sleep quality of the workers. Results The detection rates of occupational stress, job burnout and sleep disturbance among the study subjects were 33.6%, 65.4% and 23.3%, respectively. Multivariate logistic regression analysis showed that the workers with a monthly income <5 000 yuan had a higher risk of occupational stress than those with a monthly income ≥5 000 yuan (P<0.01). The workers with ≥5.0 years of service had a higher risk than those with <1.0 year (P<0.05). Lack of physical exercise, employment in medium- and large-sized enterprises, and shift work were risk factors of occupational stress in the workers (all P<0.01). The workers aged 18-<30 years had a higher risk of job burnout than those aged 45-<60 years (P<0.05). The workers monthly income <5 000 yuan was associated with a higher risk of job burnout than those with ≥9 000 yuan (P<0.05). The workers with 1.0-<10.0 years or ≥15.0 years of service had higher job burnout risks than those with <1.0 year (all P<0.05). Being unmarried, lack of physical exercise, and employment in medium- and large-sized enterprises were risk factor of job burnout in the workers (all P<0.05). The workers with an educational level of high school or above had a higher risk of sleep disturbance than those with junior school or below (P<0.05). The workers who work >56 hours per week had a higher risk than those working ≤40 hours per week (P<0.01). Conclusion There is a high detection rate of occupational stress, job burnout, and sleep disturbance in the secondary industry workers in Jinshan District, Shanghai City. Special attention should be given to workers with low income, lack of physical exercise, employment in medium- and large-sized enterprises, shift work, long service duration, and long weekly working hours to protect their physical and mental health.

Objective To establish the preliminary reference intervals for the percentage and fluorescence intensity ratio(FIR)for 12 platelet-leukocyte aggregates(PLAs)in peripheral blood circulation.Methods A total of 124 healthy individuals(61 males and 63 females)aged 18-90 years were selected from Beijing Jishuitan Hospital.Venous blood samples were collected in EDTA-K2 tubes for anticoagulation,and flow cytometry was used to measure the percentage and FIR of 12 types of PLAs.All the participants were grouped by gender for comparative analysis.Reference intervals for each parameter were calculated according to the WS/T 402-2024 standard.Results The percentages of platelet-T lymphocyte aggregates,platelet-CD4+T lymphocyte aggregates,and platelet-CD8+T lymphocyte aggregates in males were significantly lower than those in females(all P＜0.05).The FIRs of platelet-T lymphocyte aggregates,platelet CD8+T lymphocyte aggregates,platelet-B lymphocyte aggregates,and platelet-NK cell aggregates in males were significantly higher than those in females(all P＜0.05).Conclusion The preliminary reference intervals for the percentage and FIR of 12 types of PLAs in healthy adults have been established.Different gender may influence the detection results of platelet-leukocyte aggregates,especially platelet-lymphocyte aggregates,and the corresponding FIR values in healthy adults.Further large-scale studies should be necessary to confirm these findings.

BACKGROUND:It has been found that anterior knee pain is related to the biomechanics of the lower limbs,but there is still a lack of research on the effects of gluteal muscle training on the knee joint and daily activities of the lower limbs.OBJECTIVE:To investigate the effects of gluteal muscle activation exercise therapy on the muscle strength of hip and knee joint muscle groups and pain in young male patients with anterior knee pain.METHODS:Twenty-five young male patients with anterior knee pain were enrolled and randomly divided into two groups:gluteal muscle activation group(n=12)and blank control group(n=13).The gluteal muscle activation group performed gluteal muscle activation exercises,40 minutes each,3 times/week,for 6 weeks.The blank control group did not perform any intervention.Assessments were conducted at the time of enrollment and again after 6 weeks.The relative peak torque,total work,ratio of flexors and extensors,and muscle endurance values of the affected hip and knee joints were evaluated through isokinetic flexion and extension tests at 60(°)/s and 180(°)/s.At the same time,floors at which climbing was stopped in the stair-climbing test were detected and the visual analog scale score was assessed.RESULTS AND CONCLUSION:(1)Isokinetic knee extension and flexion test:For the hip joint,the gluteal muscle activation group showed a significant increase in the relative peak torque at 60(°)/s and 180(°)/s by 29.74%and 25.95%respectively after intervention(P=0.022,P=0.024);the blank control group showed a 12.12%decrease in muscle endurance at 180(°)/s compared to before intervention(P=0.000).For the knee joints,the gluteal muscle activation group had a significant increase in the relative peak torque at 60(°)/s and 180(°)/s by 18.69%and 7.27%respectively after intervention(P=0.006,P=0.033);there were no significant changes in the blank control group before and after intervention(P>0.05).(2)Stair-climbing test:The number of floors climbed to cessation in the gluteal muscle activation group was(6.41±6.1)floors,which was higher than that in the blank control group(P=0.024),and increased by 33.11%compared with before intervention(P=0.016);there were no significant changes in all the indicators of the blank control group before and after intervention(P>0.05).(3)Pain assessment:After intervention,the visual analogue scale score of the gluteal muscle activation group was significantly lower than that of the blank control(P=0.036),and also decreased compared to before intervention(P=0.000);there were no significant changes in the blank control group before and after intervention(P>0.05).To conclude,the 6-week gluteal activation exercise therapy can improve the explosive power and endurance of the lower limb muscles,and reduce the degree of anterior knee pain.For patients with anterior knee pain,gluteal muscle training is necessary to promote recovery.

BACKGROUND:It has been found that anterior knee pain is related to the biomechanics of the lower limbs,but there is still a lack of research on the effects of gluteal muscle training on the knee joint and daily activities of the lower limbs.OBJECTIVE:To investigate the effects of gluteal muscle activation exercise therapy on the muscle strength of hip and knee joint muscle groups and pain in young male patients with anterior knee pain.METHODS:Twenty-five young male patients with anterior knee pain were enrolled and randomly divided into two groups:gluteal muscle activation group(n=12)and blank control group(n=13).The gluteal muscle activation group performed gluteal muscle activation exercises,40 minutes each,3 times/week,for 6 weeks.The blank control group did not perform any intervention.Assessments were conducted at the time of enrollment and again after 6 weeks.The relative peak torque,total work,ratio of flexors and extensors,and muscle endurance values of the affected hip and knee joints were evaluated through isokinetic flexion and extension tests at 60(°)/s and 180(°)/s.At the same time,floors at which climbing was stopped in the stair-climbing test were detected and the visual analog scale score was assessed.RESULTS AND CONCLUSION:(1)Isokinetic knee extension and flexion test:For the hip joint,the gluteal muscle activation group showed a significant increase in the relative peak torque at 60(°)/s and 180(°)/s by 29.74%and 25.95%respectively after intervention(P=0.022,P=0.024);the blank control group showed a 12.12%decrease in muscle endurance at 180(°)/s compared to before intervention(P=0.000).For the knee joints,the gluteal muscle activation group had a significant increase in the relative peak torque at 60(°)/s and 180(°)/s by 18.69%and 7.27%respectively after intervention(P=0.006,P=0.033);there were no significant changes in the blank control group before and after intervention(P>0.05).(2)Stair-climbing test:The number of floors climbed to cessation in the gluteal muscle activation group was(6.41±6.1)floors,which was higher than that in the blank control group(P=0.024),and increased by 33.11%compared with before intervention(P=0.016);there were no significant changes in all the indicators of the blank control group before and after intervention(P>0.05).(3)Pain assessment:After intervention,the visual analogue scale score of the gluteal muscle activation group was significantly lower than that of the blank control(P=0.036),and also decreased compared to before intervention(P=0.000);there were no significant changes in the blank control group before and after intervention(P>0.05).To conclude,the 6-week gluteal activation exercise therapy can improve the explosive power and endurance of the lower limb muscles,and reduce the degree of anterior knee pain.For patients with anterior knee pain,gluteal muscle training is necessary to promote recovery.

Objective:To analyze the phenotypes and gene mutations of the three families with hereditary protein S (PS) deficiency caused by frameshift heterozygous variants.Methods:Case report and case series study. We investigate three probands and their family members (14 people from three generations) who registrated in Beijing Jishuitan Hospital of Capital Medical University from Feb 1st to 28th, 2025. The clinical data of the three probands and their family members were collected. The related coagulation tests of all members were detected. Protein S activity (PS:A) was determined using coagulation assay, and total protein S antigen (TPS:Ag) and free protein S antigen (FPS:Ag) were measured using ELISA. All exons and their flanking sequences of the probands were amplified using PCR technique, and gene analysis by direct sequencing, and variant genes were searched which were then verified by cloning sequencing, meanwhile, the corresponding variant loci of the family members were analyzed. A calibrated automated thrombin generation (CAT) method was used to study thrombin production. ClustalX-2.1-win software was used to analyze the conservatism of the mutant loci; Mutation Taster and Franklin.genoox online bioinformatics software were used to predict the pathogenicity of the mutant loci; PyMol software was used to analyze the changes in protein spatial structure before and after mutation.Results:The PS:A, TPS:Ag and FPS:Ag of the three probands with hereditary PS deficiency were decreased. Genetic sequencing identified a total of three PROS1 genetic variants, and all the three probands were heterozygous frameshift variants: p.Gln51Argfs*2 in Proband A; p.Met251Valfs*17 in Proband B; and Thr478tyrfs*21 in Proband C. Conservative analysis showed that p.Gln51, p.Met251 and p.Thr478 loci were not conserved among the homologous species; Mutation Taster and Franklin.genoox analysis showed that these variants were pathogenic; protein structure model analysis showed that p.Gln51Argfs*2, p.Met251Valfs*17 and p. Thr478Tyrfs*21 variants may result in altered protein spatial structure.Conclusion:The heterozygous frameshift variations of PS p.Gln51Arg fs*2, p.Met251Valfs*17 and p.Thr478Tyrfs*21 would alter the spatial structure of PS molecules, and reduce their structural stability, leading to PS deficiency.

AIM To analyze the metabolites of nobiletin in rats in vivo based on characteristic ions.METHODS Ten rats were assigned into administration group and control group,and given intragastric administration of the 0.5％CMC-Na suspension of nobiletin(250 mg/kg)and 0.5％CMC-Na solution,respectively,after which plasma,urine and feces were collected,solid phase extraction method was adopted in pretreatment,UHPLC-HRMS analysis was performed.The candidate metabolites were systematically described according to diagnostic product ions,chromatographic retention time,accurate molecular weight and neutral loss fragments,after which accurate metabolites were obtained in the established metabolite data set with-CH3(m/z 15)characteristic ions as a baits.RESULTS A total of 64 metabolites were identified,whose main metabolic pathways were glucuronidation,sulfation,hydrogenation and their compound reactions.CONCLUSION This experiment elucidates the metabolites of nobiletin in rats in vivo,which provides a new reference for its further development.

Objective To investigate the risk factors for acute calf muscular venous thrombosis(CMVT)in elderly patients with hip fractures and evaluate the diagnostic value of thrombotic risk index for CMVT.Methods The blood samples from elderly emergency admission patients with traumatic hip fractures were prospectively collected,and platelet count(PLT),plateletcrit(PCT),coagula-tion markers such as thrombin-activatable fibrinolysis inhibitor(TAFI),D-dimer,and fibrinogen(Fib),and biochemical markers,including serum total protein(TP),globulin(Glob),creatine kinase(CK),triglyceride(TG),and lactate dehydrogenase(LDH),were detected.The differences in these markers between the CMVT group,deep calf venous thrombosis(DCVT)group and non-deep venous thrombosis(DVT)group were compared.The multivariate logistic regression analysis was used to identify independent risk fac-tors for CMVT,and a thrombotic risk index was generated to evaluate its diagnostic value in CMVT.Results Compared with the non-DVT group,the levels of TAFI,Fib,TP,CK,TG,PCT,and LDH in the CMVT group were significantly reduced(P<0.05),while the level of D-dimer increased but the difference was not statistically significant.The level of TP in the DCVT group was significantly lower than that in the non-DVT group(P<0.05).Multivariate analysis showed that age,LDH,and Fib were independent risk factors for CMVT.The area under the receiver operating characteristics(ROC)curve of the thrombotic risk index in diagnosing CMVT was 0.718(P<0.001).Conclusion Age,LDH,and Fib are the independent risk factors of CMVT in elderly patients with hip fractures.The thrombotic risk index has a high diagnostic value in CMVT.