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 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 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.