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 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 study the clinical efficacy of irbesartan hydrochlorothiazide combined with small dose thyroxine in the treatment of severe heart failure in the elderly. Methods 100 elderly patients with severe heart failure were selected as the research object (from January 2013 to December 2016). They were randomly divided into the control group and the experimental group, with 50 patients in each group. Patients in the control group received routine treatment, including diuretic therapy, cardiac treatment, and anti heart failure treatment. The experimental group was treated with low-dose thyroxine combined with irbesartan and hydrochlorothiazide on this basis. The clinical effects of the two groups were compared and analyzed. Results After the corresponding treatment, the number of invalid cases in the experimental group was 6 cases, 16 cases were effective, 28 cases were effective,and the total effective number was 44 cases. In the control group, the number of invalid cases was 15. The effective rate of the treatment group was 88.0％, which was significantly higher than that of the control group 70％, which was statistically significant (P<0.05). After treatment, the left ventricular ejection fraction, the level of four and three, and the level of serum thyroid stimulating hormone in the experimental group were significantly higher than those before treatment, with statistical difference (P<0.05). Conclusion Small dose thyroxine combined with irbesartan and hydrochlorothiazide is effective in the treatment of severe heart failure in elderly patients, and can significantly improve the heart function of patients, and has the significance of further clinical promotion.

Objective To study and analyze the clinical efficacy of metoprolol combined with trimetazidine in the treatment of coronary heart disease and heart failure. Methods 80 patients with coronary heart disease and heart failure treated in our hospital from January 2015 to December 2016 were selected and randomly divided into the control group and the experimental group, with 40 cases in each group. The control group received routine treatment, including diuretics and angiotensin converting enzyme inhibitors, and the experimental group was treated with trimetazidine combined with metoprolol. The therapeutic effects of the two groups were compared and analyzed. Results After the corresponding treatment, 5 patients in the experimental group were invalid. In the control group, 14 patients were ineffective. The effective rate of treatment in the experimental group (87.5％) was significantly higher than that in the control group (65.0％), the differences were statistically significant (P<0.05). After treatment, the LVEF in the experimental group was (50.4 ± 7.7)％, and the LVEF in the control group was (45.1 ± 7.0)％. In addition, the LVEDD and LVESD indexes and LVEF in the experimental group were better than those in the control group, the differences were statistically significant (P<0.05). Conclusion The clinical effect of trimetazidine combined with metoprolol in the treatment of coronary heart disease and heart failure is better. It can improve the efficiency of treatment to some extent, and has the significance of further promotion and application.