Introduction: Vitamin K (VK) as well as vitamin D (VD) plays an important role in osteoporosis. Vitamin K1 (VK1), vitamin K2 (VK2, menaquinone-4 (MK-4), and menaquinone-7 (MK-7)) are significant for the metabolism of skeletal muscle. 25-hydroxyvitamin D2 and 25-hydroxyvitamin D3 (25(OH)D2 and 25(OH)D3) reflect circulating VD levels. More sensitive measurements remain to be developed. In the present study, a new high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed for the determination of VK1, VK2 (MK-4 and MK-7), as well as 25(OH)D2 and 25(OH)D3 levels in human serum. Methods: We developed a simple LC-MS/MS method for the determination of VK1, MK-4, MK-7, 25(OH)D2, and 25(OH)D3 levels in human serum and validated the method in a study cohort of 200 patients divided into the premenopausal women group and postmenopausal osteoporosis patient group. Results: The overall precision (coefficient of variation) ranged from 2.66 to 10.11% in the specified working range (0.05–5 ng/mL) for VK1, MK-4, and MK-7. Serum VK1, MK-4, and MK-7 levels in postmenopausal women with osteoporosis were 1.187 ± 0.094 ng/mL, 0.058 ± 0.009 ng/mL, and 0.885 ± 0.064 ng/mL, respectively (mean ± standard deviation). Serum VK1, MK-4, and MK-7 levels in premenopausal women were 1.143 ± 0.103 ng/mL, 0.028 ± 0.003 ng/mL, and 1.553 ± 0.226 ng/mL, respectively. Serum 25(OH)D2 and 25(OH)D3 levels in postmenopausal women with osteoporosis were 0.757 ± 0.056 ng/mL and 11.72 ± 0.632 ng/mL, respectively. Serum 25(OH)D2 and 25(OH)D3 levels in premenopausal were 1.793 ± 0.575 ng/mL and 12.42 ± 1.069 ng/mL, respectively. Conclusion: A new LC-MS/MS method for determination of serum VK and VD levels was evaluated and validated. MK-7 in plasma decreased earlier than VD in postmenopausal osteoporosis patients. MK-7 status is significantly associated with osteoporosis and could be considered a predictable biomarker in the diagnosis of osteoporosis in postmenopausal women.

Venous thromboembolism is a common complication following trauma. We investigated the dynamics of plasma microparticles (MPs) levels and explored their potential as biomarkers of deep vein thromboembolism (DVT) after trauma. A total of 775 patients with traumatic fractures were recruited in this nested study. About 106 trauma patients (53 DVT subjects and 53 age-, sex-, and fracture site-matched non-DVT subjects) and 53 healthy volunteers met the enrollment criteria. MPs were characterized by transmission electron microscope, nanoparticle tracking analysis, and western blotting. Circulating levels of MPs were measured using a flow cytometer. Meanwhile, routine laboratory parameters were examined in all patients. Compared to non-DVT patients, DVT patients had higher circulating phosphatidylserine (PS) + MPs, hepatocyte-derived MPs (HMPs), PS + HMPs, and platelet-derived MPs (PMPs). Notably, PS + HMPs had the best predictive value for DVT diagnosis in trauma patients (area under the curve [AUC] 0.8939, 95% CI 0.8326 to 0.9552), which was superior to d-dimer (AUC 0.5881). The Hepatic Procoagulant Index combined plasma levels of PS + HMPs and albumin, increasing the AUC to 0.8978 (95% CI 0.8396 to 0.9561). This is the first study that addressed circulating PS + HMPs are promising biomarkers with high performance in diagnosing DVT. The Hepatic Procoagulant Index is a potential predictor of DVT in trauma patients.

目的 探讨补体C3和C4与老年髋部骨折术前深静脉血栓形成(DVT)的相关性及其诊断价值.方法 连续纳入北京积水潭医院老年髋部骨折患者并观察,行术前血管造影或彩超以筛查DVT并确诊DVT的患者共50例纳入DVT组,在未确诊DVT患者中以1∶1的比例纳入与DVT组性别、年龄匹配的患者作为对照组(非DVT组).用免疫比浊法测定血清C3和C4浓度并进行统计学分析;绘制ROC曲线以评估各指标对DVT的诊断价值.结果 DVT组、非DVT组血清C3浓度分别为(1.162±0.223)g/L、(0.869±0.206)g/L;C4 浓度分别 为(0.307±0.070)g/L、(0.257±0.079)g/L,DVT 组 C3、C4 水平均高于非DVT组(P均＜0.05).ROC曲线表明,C3的ROC曲线下面积(AUCROC)为0.839,当cut-off值为0.893 g/L时,敏感性为70.0％(95％CI:56.3％～80.9％),特异性为 94.0％(95％CI:83.8％～98.4％);C4 的 AUCROC为 0.683,当 cut-off 值为 0.267 g/L 时,敏感性为 58.0％(95％CI:44.2％～70.6％),特异性为 76.0％(95％CI:62.6％～85.7％).纳入 C3、Fib、D-二聚体以及 Caprini 评分构建回归模型计算的组合因子AUCROC为0.943.结论 补体C3和C4与老年髋部骨折术前DVT相关联并具有诊断价值.

OBJECTIVE:Venous thromboembolism (VTE) is a common and severe complication of joint arthroplasty. Microparticles (MPs) containing phosphatidylserine (PS) and tissue factor (TF) can trigger coagulation in VTE. This study aims to measure and compare MP levels in joint arthroplasty patients with and without VTE. METHODS:This prospective cohort study enrolled 181 patients who underwent joint arthroplasty. Ultrasound examination was used to diagnose VTE on preoperative day 0 and postoperative day 6. MPs were analysed using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and flow cytometry. The levels of platelet-derived microparticles (PMPs), endothelial cell-derived microparticles (EMPs), granulocyte-derived microparticles (GMPs), red cell-derived microparticles (RMPs), monocyte-derived microparticles (MMPs), Annexin V+ MPs (AV+ MPs), and tissue factor+ MPs (TF+ MPs) derived from five kinds of MPs were measured on day 0 (before surgery), 1, 2, 3, 4, 5, and 6 after surgery. RESULTS:The levels of AV-TF+ EMPs and AV-TF+ MMPs were significantly increased in patients with VTE on postoperative day 5 compared to those without VTE (P = 0.031 and P = 0.031, respectively). CONCLUSION:AV-TF+ MPs may indicate the development of VTE and serve as predictive markers in joint arthroplasty patients.

Background: Deep venous thrombosis (DVT) is the most common complication in patients with traumatic fractures. The neutrophil extracellular traps (NETs) can promote thrombus formation. In this prospective study, we investigated the role of NETs in thrombosis in patients with traumatic fractures to evaluate whether the biomarkers of NETs can be used to help predict the risk of thrombosis. Methods: Traumatic fracture patients were enrolled in this prospective observational cohort study. Healthy controls (Control); patients with lower extremity fractures who neither present with nor develop DVT (Trauma non-DVT); patients with lower extremity fractures who do not present with DVT but do develop DVT (Trauma DVT); and patients with lower extremity fractures who present with DVT (DVT) were included. NETs biomarker levels of Citrullinated Histone H3 (H3Cit), cell-free DNA (cfDNA) and nucleosomes in the plasma were determined. The D-dimer and fibrin(-ogen) degradation products (FDP) level in plasma was measured. Statistical analysis of the test results was performed to assess changes in NETs biomarker levels during thrombosis in patients with traumatic fractures. Results: The H3Cit levels in the DVT group were significantly greater than in the Trauma non-DVT group and Trauma DVT group (1.88(1.11, 3.35) ng/ml Vs 0.38(0.10, 1.17) ng/ml, P <= 0.05). The level of cfDNA was significantly greater in patients with traumatic fractures and was higher after thrombosis than before. The levels of D-dimer in the DVT, Trauma DVT, and Trauma non-DVT groups were significantly greater than in the Control group (5.11(3.97, 8.11) mg/l; 6.12(2.59, 18.49) mg/l; 2.99(0.99, 9.02) mg/l Vs 0.18(0.08,0.24) mg/l, P < 0.05). The distribution of FDP levels in each group was similar to that of D-dimer. Data are presented as medians (25th percentile, 75th percentile). Conclusions: NETs released by neutrophils are involved in the formation of DVTs in patients with traumatic fractures. H3Cit and cfDNA can assist the diagnosis of DVT in patients with traumatic fractures.

Objective Deep venous thrombosis (DVT) is a common complication in patients with traumatic injury. Tissue factor pathway inhibitor (TFPI) is a natural anticoagulant protein in the extrinsic coagulation pathway. However, the relationship between DVT after trauma and the anticoagulant activity of TFPI remains unclear. In this prospective study, we investigated the role of TFPI in trauma patients with DVT to evaluate whether the anticoagulant activity of TFPI measured by a new functional assay can be used to help predict the risk of DVT. Patients and methods: This prospective nested case-control study enrolled trauma patients and healthy volunteers. Forty-eight trauma patients diagnosed with DVT and forty-eight matched trauma patients without DVT were included in the study. 120 healthy volunteers were also included as controls. Blood samples and case information were collected at admission. Patients accepted angiography before surgery to diagnose DVT. The parameters examined included TFPI anticoagulant activity, free-TFPI antigen, blood cell counts, and routine clinical coagulation tests. Results: For the parameters of TFPI anticoagulant activity, three were markedly increased in the DVT group compared to the non-DVT group (TFPI initial anticoagulant time ratio, P = .022; TFPI whole anticoagulant time ratio, P = .048; and TFPI anticoagulant rate, P = .034). The free-TFPI antigen concentration also showed a significant increasing trend in trauma patients with DVT compared with trauma patients without DVT (P = .035). Multivariate logistic regression analysis identified four independent factors for the development of DVT (TFPI initial anticoagulant time ratio, free-TFPI antigen, prothrombin time, and red blood cell count). We calculated the TFPI correlation coefficient and found that the area under the receiver operating characteristic curve was .821. Conclusions: A novel functional assay was developed to measure the anticoagulant activity of TFPI. The anticoagulant activity of TFPI can be used as a potential biomarker for diagnosing DVT in trauma patients.

Effective and safe hemodialysis is essential for patients with acute kidney injury and chronic renal failures. However, the development of effective anticoagulant agents with safe antidotes for use during hemodialysis has proven challenging. Here, we describe DNA origami-based assemblies that enable the inhibition of thrombin activity and thrombus formation. Two different thrombin-binding aptamers decorated DNA origami initiates protein recognition and inhibition, exhibiting enhanced anticoagulation in human plasma, fresh whole blood and a murine model. In a dialyzer-containing extracorporeal circuit that mimicked clinical hemodialysis, the origami-based aptamer nanoarray effectively prevented thrombosis formation. Oligonucleotides containing sequences complementary to the thrombin-binding aptamers can efficiently neutralize the anticoagulant effects. The nanoarray is safe and immunologically inert in healthy mice, eliciting no detectable changes in liver and kidney functions or serum cytokine concentration. This DNA origami-based nanoagent represents a promising anticoagulant platform for the hemodialysis treatment of renal diseases.

ObjectivesImplant-related infection is a common complication after orthopedic surgery, but there is limited research focused on anaerobic infections. We retrospectively analyzed data from 80 patients with anaerobic implant-related infections in order to investigate the clinical features, bacterial distribution and antimicrobial resistant characteristics of this disease.Methods80 patients who underwent implant-related infections with anaerobes were included. Pathogens were isolated and identified by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry with verification of 16s rRNA sequencing. Antimicrobial susceptibility testing (AST) was performed using Epsilometric test (E-test).ResultsAmong the 80 patients, 61.2% (49/80) were infected with anaerobes alone, while 38.8% (31/80) were co-infected with anaerobes and other bacteria. Early infection cases involving anaerobe-alone infections were significantly higher compared to the co-infection group (P < 0.001), also exhibiting lower levels of neutrophils (P = 0.033) and ESR (P = 0.046). Anaerobe-alone infections in the prosthetic joint infection group represented a higher proportion compared with other implant-related infections (P = 0.031). Among all species of anaerobes identified, the top 3 were Cutibacterium acnes, Finegoldia magna and Peptostreptococcus anaerobius. Low MIC values to vancomycin was recorded in C. acnes strains and for amoxicillin/clavulanic acid and piperacillin/tazobactam in most F. magna strains. One of the C. acnes and F. magna strains appeared multi-drug resistant except to vancomycin.ConclusionsAnaerobe-alone infections have later first onset times and lower infection biomarker levels compared to co-infected patients. The first choice against C. acnes is vancomycin, while amoxicillin/clavulanic acid and piperacillin/tazobactam are recommended for F. magna.

目的 调查不同妊娠期、不同年龄孕妇体内维生素E(VE)营养状态,探讨 α-生育酚作为评价指标的适用性.方法 回顾性研究,选取2016年10月至2017年10月期间在北京积水潭医院回龙观院区产科规律产检的孕妇,1840例孕妇均遵医嘱在妊娠早、中、晚期进行血液采集,采用高效液相色谱法(HPLC)检测血清中 α-生育酚水平.结果 α-生育酚水平随孕周增加而增加(F=6409.353,P<0.001),同时VE增多率也由妊娠早期的0.33％、妊娠中期的9.51％,增至妊娠晚期的34.18％;在妊娠早、中期,孕妇年龄越大 α-生育酚水平越高(F=11.376、P<0.001,F=6.282,P<0.001),在妊娠晚期虽然差别不具有统计学意义(F=2.343、P=0.071),但是随着孕妇怀孕年龄的增大,α-生育酚水平有增高的趋势.结论 北京市昌平地区的孕妇在妊娠期间VE水平按目前的分层标准,极少存在缺乏的表现,异常主要表现为VE增多.α-生育酚水平随孕周增加而增加,怀孕年龄越大 α-生育酚水平越高.但是在分析临床检测数值时,应该充分考虑年龄、特别是妊娠期间血脂生理性增多对循环中 α-生育酚水平的影响,审慎做出VE过剩的结论.

Deep vein thrombosis (DVT) is a common complication after trauma. The development of markers to predict DVT in trauma patients is needed, and circulating microparticles (MPs) and their contents are possible candidates. In this study, we aimed to identify platelet factor 4 (PF4) and β-thromboglobulin (β-TG) mRNAs in circulating MPs as potential markers for DVT diagnosis in trauma patients. Fifteen trauma patients diagnosed with DVT and fifteen matched patients without DVT were included in this study. Fifteen healthy volunteers also were included as controls. Circulating MPs were obtained from the plasma of all study subjects. Annexin V+ MPs and platelet-derived MPs (PMPs) were quantified using flow cytometry. PF4 and β-TG mRNAs in MPs were determined by qPCR, and the common logarithm of relative quantitation (RQ) was calculated using the comparative Ct method. Receiver-operating characteristic (ROC) curves were performed to analyze the diagnostic value of PF4 and β-TG mRNAs. No significant differences were found in Annexin V+ MPs and PMPs levels between trauma patients with and without DVT. However, both PF4 and β-TG mRNAs in MPs from the DVT group were significantly higher than the non-DVT group and healthy controls (P = 0.014 for PF4, P = 0.010 for β-TG). The ROC curve analysis showed that both the PF4 mRNA (area-under curve (AUC) 0.756, P = 0.017) and the β-TG mRNA (AUC 0.751, P = 0.019) had a positive predictive value for DVT. This finding indicates that the PF4 and β-TG mRNAs in MPs may be used as potential biomarkers for DVT diagnosis in trauma patients.

To evaluate variations in coagulation, fibrinolysis and endothelial marker expression in cirrhotic patients and to explore their clinical value and predictive performance in cirrhotic patients with or without portal vein thrombosis (PVT), we performed a case-control study with 175 cirrhotic patients and 50 healthy individuals. 99 patients had PVT and another 76 patients did not. All participants were evaluated for plasma levels of conventional hemostatic markers. Thrombin-antithrombin complex (TAT), plasmin-α2-plasmin inhibitor complex (PIC), thrombomodulin (TM), tissue plasminogen activator inhibitor complex (t-PAIC), von Willebrand factor antigen (vWF: Ag) and coagulation factor Ⅷ (FⅧ: c) were also assessed and the ratio of TAT/t-PAIC was calculated. We analyzed differences in these biomarkers among the three groups and constructed receiver operating characteristic (ROC) curves. Patients with PVT exhibited significantly higher TAT and TAT/t-PAIC than cirrhotic patients without PVT (both P < 0.001). Areas under the curve (AUC) of ROC analyses for TAT and TAT/t-PAIC were 0.68 and 0.66, the cut-off levels were 1.55 ng/ml and 0.46, with sensitivities and specificities of 78.79% and 51.32% regarding TAT, 39.8% and 90.79% regarding TAT/t-PAIC. Levels of FⅧ: c and vWF: Ag in patients with PVT were significantly lower than those without PVT (p = 0.026 and p = 0.027, respectively). The AUCROC, cut-off level, sensitivity and specificity of FⅧ: c were 0.64, 111.1%, 66.67% and 60%, respectively. For vWF: Ag they were 0.61, 429%, 89.66% and 38.71%, respectively. Cirrhotic patients have disorders of coagulation, fibrinolysis and the endothelial system. TAT, TAT/t-PAIC, FⅧ: c and vWF: Ag can be used as potential biomarkers for predicting PVT in cirrhotic patients.

Background: Deep vein thrombosis (DVT) is a common complication in patients with traumatic injury. The purpose of this study was to develop a potential predictor of DVT. Methods: This case-control study enrolled adult trauma patients and healthy volunteers. Patients underwent angiography before surgery to diagnose DVT. Patients with or without DVT were matched by gender, age and fracture sites. Laboratory parameters included lysis potential (LP), lysis time (LT), blood cell counts, conventional coagulation tests, tissue plasminogen activator inhibitor complex (tPAIC) and others. Results: 41 of 319 patients with DVT were matched with 41 patients without DVT and 80 healthy volunteers were controls. LP and LT were significantly decreased in patients with DVT than without (P = 0.043 and P = 0.014, respectively). The level of tPAIC in the DVT group was significantly higher than in patients without DVT (P = 0.042). We defined the Fibrinolysis Index as (-10.707) x LP + (-0.607) x LT (min) + 0.012 x fibrinogen (mg/dl) + 0.299 x tPAIC (ng/ml) + 9.917, and found that the area under the receiver operating characteristic curve for the Fibrinolysis Index was 0.802, making it a novel indicator. Conclusion: The Fibrinolysis Index represents a new discriminator for predicting DVT after traumatic lower extremity fractures.

目的 调查北京市昌平地区不同妊娠期、不同年龄段孕妇血清维生素A(VA)的营养状态,为妊娠期合理补充维生素提供参考.方法 选择2016年10月至2017年10月在北京积水潭医院回龙观院区产科规律产检的孕妇1840人,分别在其妊娠早、中、晚期三个时间段采集血液,用高效液相色谱法(HPLC)检测血清中VA水平.结果 孕妇血清VA水平在妊娠晚期下降明显,低于妊娠早期和中期(F=711.916,P<0.001),VA缺乏及边缘型缺乏率由妊娠早期的3.86％、妊娠中期的2.39％,增至妊娠晚期的18.48％.在妊娠早期,≤30岁孕妇血清VA水平低于>30岁孕妇,差异有统计学意义(t=-6.290,P<0.001).≤30岁孕妇在妊娠早期发生VA缺乏及边缘型缺乏的比例(4.79％)高于>30岁孕妇(2.64％),差异有统计学意义(χ2=6.153,P=0.046).结论 北京市昌平地区的孕妇在妊娠期间VA水平异常主要表现为以边缘型缺乏为主,而妊娠晚期VA缺乏及边缘型缺乏率增加需加强重视,对缺乏状况予以及时纠正;妊娠早期要加强对≤30岁孕妇血清VA营养水平的监测.

One common complication after joint arthroplasty is venous thromboembolism (VTE). Therefore, it is essential to measure the changes in coagulation and fibrinolysis in order to predict VTE among patients who underwent joint arthroplasty. This study aimed to identify potential useful biomarkers for prognosing to VTE. This was a prospective cohort study enrolling 83 patients who underwent joint arthroplasty. The levels of d -dimer, thrombin–antithrombin complex (TAT), plasmin–α2-antiplasmin complex (PIC), soluble thrombomodulin, and tissue plasminogen activator inhibitor complex were measured on day 0 (before surgery) and days 1, 3, and 6 after surgery. Ultrasound examination was used to diagnose VTE on preoperative day 0 and postoperative day 6. A total of 35 patients developed VTE after surgery. Patients with VTE exhibited significantly higher levels of d -dimer and TAT on postoperative days 3 and 6 (all P < .05). The area under curves (AUC) of receiver operating characteristic (ROC) were 0.65 and 0.68 and 0.68 and 0.74 for d -dimer and TAT levels on postoperative days 3 and 6, respectively. The level of TAT/PIC ratio on postoperative day 6 was significantly increased among patients with VTE compared to non-VTE patients ( P < .0001). In addition, the AUC of ROC, cutoff level, sensitivity, specificity, positive-predictive value, and negative-predictive value of TAT/PIC ratio were 0.78, 4.03 ng/TU, 97.14%, 33.33%, 51.52%, and 94.12%, respectively. The high sensitivity and negative predictive value of TAT/PIC ratio make it a potential prognostic index for diagnosing VTE during the early phase of postoperative joint arthroplasty.