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Abstract Background R189W and K193del of protein C (PC) were hotspot mutations in Chinese population with venous thromboembolism (VTE), but almost two-thirds of patients with above mutations coexisting with other genetically or aquiredly prothrombotic risk factors. The aim of this study is to clarify the independent contributions of R189W or K193del to VTE risk. Methods 490 unrelated patients with a personal history of VTE and 410 healthy participants were enrolled in this study. Data of their demographics, family history, genetic and acquired thrombosis risk factors were collected and statistically analyzed. Results PC R189W and K193del were identified in 3/410 (0.7%) and 7/410 (1.7%) healthy controls, and in 27/490 (5.5%) and 43/490 (8.8%) patients with VTE, respectively. Notably, about 70% of these mutant carriers combined with other genetic or acquired thrombophilic factors. After adjustment for age, gender, other inherited and acquired risk factors, we demonstrated that R189W and K193del were associated with 5.781-fold and 4.365-fold increased risk of VTE, respectively, which were significantly lower than the prothrombotic risk of anticoagulant deficiencies induced from rare mutations. Independent R189W or K193del mutation was not associated with earlier first-onset age as well as higher recurrent rate of VTE. However, combination of other genetic or acquired thrombophilic factors had supra-additive effects on those consequences. The more additional risk factors the patients had, the younger first-onset ages and higher risk of recurrence would be. Conclusions As the most frequent mutations for PC deficiency in Chinese population, both R189W and K193del mutations had limited independent contributions to VTE development compared with other rare mutations in PROC gene, but may act in concert with other genetic defects or acquired thrombotic risk factors to produce the final severe phenotype.

Abstract Background Factor VII deficiency is a rare bleeding disorder caused by a deficiency of clotting factor VII. However, there have been some case reports of venous thrombosis in patients with factor VII deficiency, especially underlying the prothrombotic risk factors exposure. Patients with factor VII deficiency require special considerations before undergoing surgery to minimize the risk of bleeding or thrombogenesis. Case presentation Here, we described a patient with early-stage thymoma and severe factor VII deficiency who experienced an unprovoked thrombotic episode before thymectomy and a fatal thrombotic event after surgery. By adopting gene screening, a reported homozygous F7 mutation (p.His408Gln) and a novel heterozygous PROS1 mutation (p.Pro147Ala) were identified. The former resulted in severe factor VII deficiency but did not protect against thrombosis, and the latter was correlated with normal expression and cofactor activities of protein S through the thrombin generation test. The perioperative infusion of recombinant factor VII concentrate and the absence of antithrombotic prophylaxis may collectively contribute to her fatal thrombotic event after surgery. Conclusions For the patients with severe factor VII deficiency undergoing surgery, uniform replacement therapy may not be recommended, and antithrombotic prophylaxis should be used in the case with thrombotic history to minimize the risk of bleeding and thrombogenesis.

Abstract Background High-cost, time-consuming and complex processes of several current approaches limit the use of noninvasive prenatal diagnosis (NIPD) for monogenic disorders in clinical application. Thus, a more cost-effective and easily implementable approach is required. Methods We established a low-cost and convenient test to noninvasively deduce fetal genotypes of the mutation and single nucleotide polymorphisms (SNPs) loci by means of targeted amplification combined with deep sequencing of maternal genomic and plasma DNA. The sequential probability ratio test was performed to detect the allelic imbalance in maternal plasma. This method can be employed to directly examine familial pathogenic mutations in the fetal genome, as well as infer the inheritance of parental haplotypes through a group of selected SNPs linked to the pathogenic mutation. Results The fetal mutations in 17 families with different types of monogenic disorders including hemophilia A, von Willebrand disease type 3, Duchenne muscular dystrophy, hyper-IgM type 1, glutaric acidemia type I, Nagashima-type palmoplantar keratosis, and familial exudative vitreoretinopathy were identified in the study. The mutations included various forms: point mutations, gene inversion, deletions/insertions and duplication. The results of 12 families were verified by sequencing of amniotic fluid samples, the accuracy of the approach in fetal genotyping at the mutation and SNPs loci was 98.85% (172/174 loci), and the no-call rate was 28.98% (71/245 loci). The overall accuracy was 12/12 (100%). Moreover, the approach was successfully applied in plasma samples with a fetal fraction as low as 2.3%. Conclusions We have shown in this study that the approach is a cost-effective, less time consuming and accurate method for NIPD of monogenic disorders.

One important limitation for achieving therapeutic expression of human factor VIII (FVIII) in hemophilia A gene therapy is inefficient secretion of the FVIII protein. Substitution of five amino acids in the A1 domain of human FVIII with the corresponding porcine FVIII residues generated a secretion-enhanced human FVIII variant termed B-domain-deleted (BDD)-FVIII-X5 that resulted in 8-fold higher FVIII activity levels in the supernatant of an in vitro cell-based assay system than seen with unmodified human BDD-FVIII. Analysis of purified recombinant BDD-FVIII-X5 and BDD-FVIII revealed similar specific activities for both proteins, indicating that the effect of the X5 alteration is confined to increased FVIII secretion. Intravenous delivery in FVIII-deficient mice of liver-targeted adeno-associated virus (AAV) vectors designed to express BDD-FVIII-X5 or BDD-FVIII achieved substantially higher plasma FVIII activity levels for BDD-FVIII-X5, even when highly efficient codon-optimized F8 nucleotide sequences were employed. A comprehensive immunogenicity assessment using in vitro stimulation assays and various in vivo preclinical models of hemophilia A demonstrated that the BDD-FVIII-X5 variant does not exhibit an increased immunogenicity risk compared to BDD-FVIII. In conclusion, BDD-FVIII-X5 is an effective FVIII variant molecule that can be further developed for use in gene- and protein-based therapeutics for patients with hemophilia A.

It is well known that canine factor VIII (cFVIII) has a higher specific activity than does human FVIII (hFVIII), and it has been previously demonstrated that cFVIII light chain is able to enhance hFVIII activity. The goal of this study was to first determine which amino acids in cFVIII light chain were responsible for enhancing hFVIII activity, and second to use these amino acids to develop a hFVIII variant with enhanced functional activity. We systemically screened segments of cFVIII light chain by testing an array of human-canine light chain hybrids and found that canine amino acids 1857–2147 were key to this enhancement. Each canine amino acid within this span was screened individually using a negative selection method, which led to the identification of 12 aa (JF12) in the FVIII light chain that could enhance activity. Substitution of the corresponding 12 aa into hFVIII (hFVIIIJF12BDD) elevated the specific activity profile in vitro. Furthermore, hFVIIIJF12BDD expressed an in vivo-displayed increased coagulation activity compared to wild-type, while maintaining normal secretion efficiency. In conclusion, we identified the amino acids in cFVIII that are the key determinants for higher specific activity and may be the basis for future development of therapeutic treatments for hemophilia A.

Abstract Background Inherited Factor XIII deficiency (FXIIID) is one of the most severe and under-diagnosed rare bleeding disorders. Only 5 large deletions involving one or more exons in F13A1 have been reported, and lacking of multiplex ligation-dependent probe amplification (MLPA) assay might underestimate the copy number variations (CNVs) in F13A1 and F13B. We had characterized the clinical presentation of two unrelated severe FXIIID probands and explored the pathogenic mechanisms. Results Both probands experienced several episodes of fatal bleeding and delayed wound healings prior to diagnosis. FXIII activity was measured by the ammonia release assay, and FXIII-A and FXIII-B antigens were determined by ELISA. All the exons including exon-intron boundaries and promoter regions of F13A1 and F13B were amplified and directly sequenced. Copy number variations (CNVs) of F13A1 and F13B were detected by the CNVplex® method. Breakpoints of the F13A1 large deletion were identified by quantitative primer walking combined long-range PCR (LR-PCR) strategies. Proband 1 was found to have compound heterozygous mutations of a novel small deletion (c.1147del) and a missense mutation p.Arg383Ser. Proband 2 was compound heterozygous for a novel large deletion (g.[77815_112815del;112837_116628del]) and a missense mutation p.Arg716Gly in F13A1. Bioinformatics analysis of the large deletion breakpoints predicted that two fork stalling and template switching and/or microhomology-mediated break-induced replication (FoSTeS/MMBIR) events with two homologies of TCT and C might be responsible for the complex rearrangement. Prophylactic replacement therapy was immediately administered for the two probands upon establishment of the diagnosis. Conclusions We detected two type I FXIIID pedigrees and adopted CNVplex® method to detect CNVs of F13A1 and F13B for the first time. A large heterozygous deletion of g.[77815_112815del;112837_116628del] in F13A1, mediated by two FoSTeS/MMBIR events, was identified.

The changes in the coagulation, fibrinolytic, and endothelial functions are correlated with the pathophysiology of the thromboembolic diseases during acute illness. However, these changes in patients with hereditary thrombophilia who were not in the acute stage of venous thromboembolism (VTE) are unclear. A panel of 4 biomarkers, including thrombin–antithrombin complex (TAT), plasmin-α2-plasmin inhibitor complex (PIC), tissue-type plasminogen activator/plasminogen activator inhibitor-1 complex (t-PAIC), and soluble thrombomodulin (sTM), were assayed in 100 healthy controls and 100 patients with thrombophilia. Although significantly higher concentrations of TAT, PIC, t-PAIC, and sTM were observed in patients with thrombophilia than in healthy controls, 70 patients showed absolutely normal levels of the above 4 biomarkers. Among the other 30 patients who had at least 1 biomarker out of the corresponding reference interval, 26 of them presented elevated PIC with or without increased TAT. Except for sTM, other 3 biomarkers did not show significant differences in patients with previous VTE compared to those without. Patients with single episode of VTE had obviously lower t-PAIC than those with multiple episodes of VTE, whereas the levels of TAT, PIC, and sTM were unassociated with the number of thrombosis episodes. Most thrombophilia patients who were not in the acute stage of VTE showed normal coagulation, fibrinolytic, and endothelial functions. Thus, we were unable to show that the one-time response of this panel was clinically helpful in determining thrombosis risk in thrombophilia individuals. Future studies should focus on the dynamic monitoring during the chronic phase of VTE to offer special advantages for patients with thrombophilia.

Abstract Background The contribution of moderate coagulation factor XII (FXII) deficiency to development of thromboembolism is still undetermined. We have tried to show the relevance of FXII deficiency to incidences of venous thrombosis by exploring the prevalence of F12 gene mutations in Chinese patients with thrombotic disorders. Methods One hundred and six patients with venous thromboembolism (VTE) and 220 healthy controls were enrolled in study. The coding region and flanking sequences of F12 gene were amplified and sequenced to identify genetic variances. Patients with F12 mutations were also screened for other thrombotic risk factors. Results Heterozygous F12 gene mutations were identified in 6 individuals with VTE and 10 healthy controls. Q336X and R66W were found in two healthy individuals; D291E was identified in a patient with DVT; and A343P was a recurrent mutation with a prevalence of 4.7% (5/106) in patient group and 3.6%(8/220) in healthy control. The prevalence of heterozygous mutations between the two groups had no significant difference. The association of A343P mutations with VTE was weak with an OR of 1.31 (95% CI 0.42-4.11). No other thrombophilia risk factors screened were positive in patients harboring heterozygous F12 mutations. Conclusions There were conflicting theories about the relationship between FXII deficiency and thrombosis formation. Heterozygous F12 mutation decreases the plasma FXII activity approximately by half and cause moderate FXII deficiency. Although multiple mutations were identified in both groups, the link between F12 heterozygous mutation and development of thrombotic disorders is weak and further studies are warranted to clarify their relationship.

Recombinant adeno-associated vectors (rAAV) are commonly purified by either chromatography or equilibrium CsCl gradient. Nevertheless, even after purification various cellular proteins often associate with rAAV vector capsids. Such co-purifying cellular proteins may raise concern about safety of gene therapy. Here we report identification and characterization of the co-purifying cellular protein in the vector preparations by using a combination of two proteomics approaches, GeLC-MS (gel electrophoresis liquid chromatography-mass spectrometry) and 2DE (two-dimensional gel electrophoresis). Most prominent bands revealed by Coomassie Blue staining were mostly similar to the AAV capsid proteins. Posttranslational modifications of capsid proteins were detected by the proteomics analysis. A total of 13 cellular proteins were identified in the rAAV vectors purified by two rounds of cesium chloride gradient centrifugation, including 9 by the GeLC-MS analysis and 4 by the 2DE analysis. Selected cellular proteins were verified by western blot. Furthermore, the cellular proteins could be consistently found associated with different AAV serotypes and carrying different transgenes. Yet, the proteins were not integral components of the viral capsis since a stringent washing procedure by column purification could remove them. These co-purified proteins in AAV vector preparations may have a role in various stages of the AAV life cycle.

Background: Whether there is a causal relationship between childhood obesity and increased risk of chronic kidney disease (CKD) remains controversial. This study sought to explore how body size in childhood and adulthood independently affects CKD risk in later life using a Mendelian randomization (MR) approach., Methods: Univariate and multivariate MR was used to estimate total and independent effects of body size exposures. Genetic associations with early-life and adult body size were obtained from a genome-wide association study of 453,169 participants in the U.K. Biobank, and genetic associations with CKD were obtained from the CKDGen and FinnGen consortia., Results: A larger genetically predicted early-life body size was associated with an increased risk of CKD (odds ratio [OR], 1.27; 95% confidence interval [CI], 1.14 to 1.41; P =1.70E-05) and increased blood urea nitrogen (BUN) levels (β=0.010; 95% CI, 0.005 to 0.021; P =0.001). However, the association between the impact of early-life body size on CKD (OR, 1.12; 95% CI, 0.95 to 1.31; P =0.173) and BUN level (β=0.001; 95% CI, -0.010 to 0.012; P =0.853) did not remain statistically significant after adjustment for adult body size. Larger genetically predicted adult body size was associated with an increased risk of CKD (OR, 1.37; 95% CI, 1.21 to 1.54; P =4.60E-07), decreased estimated glomerular filtration rate (β=-0.011; 95% CI, -0.017 to -0.006; P =5.79E-05), and increased BUN level (β=0.010; 95% CI, 0.002 to 0.019; P =0.018)., Conclusion: Our research indicates that the significant correlation between early-life body size and CKD risk is likely due to maintaining a large body size into adulthood.

Background:  Venous thromboembolism (VTE) is predisposed by thrombotic mutations in patients with hereditary thrombophilia. Although prothrombin deficiencies caused by homozygous or compound heterozygous mutations are associated with bleeding diathesis, rare cases have shown a correlation between heterozygous prothrombin mutations and thrombosis., Materials and Methods:  We surveyed genetic variants involved in thrombosis and hemostasis in 347 patients with unprovoked VTE or having a positive family history of thrombosis. For patients identified with heterozygous prothrombin mutations, we conducted family investigations and performed a thrombin generation test (TGT) to elucidate the thrombotic risk. Novel mutants were expressed and subjected to functional assays to clarify the underlying thrombotic mechanisms., Results:  Heterozygous prothrombin mutations were identified in 3.5% of patients (12/347), including three novel mutations Phe382Ser, Phe382Leu, and Asp597Tyr found in one patient each, as well as previously reported Arg541Trp mutation in four patients and Arg596Gln mutation in five patients. A total of 42 mutation carriers were identified within the 12 pedigrees, among whom 64.3% (27/42) had experienced thrombotic events. TGT results demonstrated hypercoagulability for carriers of the five mutations, with Arg596Gln showing the highest thrombin generation potential followed by Arg541Trp. The Phe382-associated mutations severely impaired thrombomodulin-binding ability of thrombin, resulting in obviously reduced protein C (PC) activation. The Asp597Tyr mutation exhibited a mild reduction in both inactivation by antithrombin and PC activation reactions., Conclusion: The presence of heterozygous prothrombin mutations represents a potential genetic predisposition for VTE. All thrombosis-associated mutations potentiate coagulation activity by either conferring antithrombin resistance and/or impairing PC pathway activity., Competing Interests: None declared., (Thieme. All rights reserved.)

Purpose: Digital breast tomosynthesis (DBT) has been introduced more than a decade ago. Studies have shown higher breast cancer detection rates and lower recall rates, and it has become an established imaging method in diagnostic settings. However, full-field digital mammography (FFDM) remains the most common imaging modality for screening in many countries, as it delivers high-resolution planar images of the breast. To combine the advantages of DBT with the faster acquisition and the unique in-plane resolution capabilities known from FFDM, a system concept was developed for application in screening and diagnosis., Approach: The concept comprises an X-ray tube with adaptive focal spot position based on the flying focal spot (FFS) technology and optimized X-ray spectra. This is combined with innovative algorithmic concepts for tomosynthesis reconstruction and synthetic mammograms (SMs)., Results: An X-ray tube with FFS was incorporated into a DBT system that performs 50-deg wide tomosynthesis scans with 25 projections in 4.85 s. Laboratory evaluations demonstrated significant improvements in the effective modular transfer function (eMTF). The improved eMTF as well as the effectiveness of the algorithmic concepts is shown in images from a clinical evaluation study., Conclusions: The DBT system concept enables high spatial resolution at short acquisition times. This leads to improved microcalcification visibility, reduced risk of motion artifacts, and shorter breast compression times. It shifts the in-plane resolution of DBT into the high-resolution range of FFDM. The presented technology leap might be a key contributor to facilitating the paradigm shift of replacing FFDM with DBT plus SM., (© 2024 The Authors.)

Introduction: In this study, we aimed to investigate the incidence of objective and subjective indicators of the prostaglandin-associated periorbital syndrome (PAPS) after continuous instillation of topical prostaglandin analogues (PGAs) in primary open-angle glaucoma or ocular hypertension patients., Methods: A self-controlled and prospective study of PGA instillation was performed in patients ( n  = 55) with primary open-angle glaucoma or ocular hypertension. Bilateral instillation of bimatoprost, travoprost, latanoprost, or tafluprost was conducted (treatment, 3-6 months). The objective indicators recorded included interpupillary distance (IPD) and exophthalmos; subjective indicators were assessed via colour pictures of the periocular area. Data from before the administration of medication served as controls. Posttreatment changes in IPD, exophthalmos, deepening of the upper eyelid sulcus, periorbital hyperpigmentation and eyelash growth were analysed., Results: Compared with those before treatment, the interpupillary distance (IPD) differed from the baseline value at 1 month after treatment ( P  < 0.0001), and the exophthalmos only significantly differed from the baseline value at month 3 ( P  = 0.0005). Visible periorbital changes at 1, 3, and 6 months after treatment were assessed, and the incidence of eyelash growth and thickening was 7.27%, 45.45% and 66.67%, respectively. The incidence of periorbital hyperpigmentation was 7.2%, 18.18% and 33.33%, respectively. The incidence of upper-eyelid sulcus deepening was 3.64%, 7.27% and 16.27%, respectively. Bimatoprost had the highest incidence of PAPS, followed by travoprost and tafluprost, and latanoprost had the lowest incidence after three months of treatment in the between-group comparison., Conclusion: As an objective index to evaluate PAPS, the change of IPD was more obvious than the exophthalmos. Visible periorbital changes gradually appeared after three months of medication. Bimatoprost caused the most severe PAPS, and latanoprost caused the least severe PAPS., Trial Registration: The study was registered at www.chictr.org.cn on 15 April 2021, under the identifier ChiCTR2100045465.

Background: The objective of this study was to develop and validate a machine learning (ML) model for predict in-hospital mortality among critically ill patients with congestive heart failure (CHF) combined with chronic kidney disease (CKD)., Methods: After employing least absolute shrinkage and selection operator regression for feature selection, six distinct methodologies were employed in the construction of the model. The selection of the optimal model was based on the area under the curve (AUC). Furthermore, the interpretation of the chosen model was facilitated through the utilization of SHapley Additive exPlanation (SHAP) values and the Local Interpretable Model-Agnostic Explanations (LIME) algorithm., Results: This study collected data and enrolled 5041 patients on CHF combined with CKD from 2008 to 2019, utilizing the Medical Information Mart for Intensive Care Unit. After selection, 22 of the 47 variables collected post-intensive care unit admission were identified as mortality-associated and subsequently utilized in the development of ML models. Among the six models generated, the eXtreme Gradient Boosting (XGBoost) model demonstrated the highest AUC at 0.837. Notably, the SHAP values highlighted the sequential organ failure assessment score, age, simplified acute physiology score II, and urine output as the four most influential variables in the XGBoost model. In addition, the LIME algorithm explains the individualized predictions., Conclusions: In conclusion, our study accomplished the successful development and validation of ML models for predicting in-hospital mortality in critically ill patients with CHF combined with CKD. Notably, the XGBoost model emerged as the most efficacious among all the ML models employed.

Collective actions aimed at achieving goals such as resource sustainability and environmental protection often face conflicting interests between individuals and groups. These social dilemmas can be modeled using public goods games and collective risk dilemmas. However, in reality, multiple generations share a common pool of resources, leading to high costs for today's overexploitation that impacts future generations' welfare. This delayed effect creates a multigenerational conflict. To address this, we develop a coupled social-ecological coevolutionary model by establishing a relationship between individual payoffs and regional environmental quality. Our goal is to study how cooperative behaviors spread in a public goods game with environmental feedback and to identify the factors influencing this spread. We achieve this by examining the mechanisms behind certain phases and phase transitions, monitoring the spatial distribution of strategies, and assessing the environmental quality of all regions. Our findings reveal some counterintuitive results. For instance, despite cooperators' ability to enhance the environment, the overall level of cooperation in the system sometimes decreases. This is linked to cooperative clusters being invaded by defectors within the clusters' cracks. Additionally, the destructive power of defection and the cost of cooperation have more complex effects on the system., (© 2024 Author(s). Published under an exclusive license by AIP Publishing.)

Chediak-Higashi syndrome (CHS) is a rare autosomal recessive genetic disorder characterized by severe immunodeficiency, albinism and coagulation deficiency. Mostly diagnosed in early childhood, this devastating condition is associated with lysosomal abnormalities attributed to the absence or impaired function of lysosomal trafficking regulator caused by mutations in the CHS1/LYST gene. In current study, we report a case of late-onset CHS caused by two novel compound heterozygous CHS1/LYST mutations: c.8407C > T, leading to early termination of translation at residue Gln2803 (p. Gln2803Ter), and a small deletion c. 4020&#95;4031del, resulting in an in-frame deletion of three amino acid residues (p. Asp1343&#95;Val1346del). Both variants retain a large part of the CHS/LYST protein, particularly p. Asp1343&#95;Val1346del, which preserves critical functional BEACH and WD40 domains in the C terminal, potentially maintaining residual activity and alleviating patient symptoms. The timeline of SARS-CoV-2 infection and rapid symptom progression suggests that the viral infection may have trigger the accelerated phase development leading to a poor prognosis., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Purpose: The study aimed to explore potential discrepancies in contrast sensitivity in the ON and OFF visual pathways among individuals with amblyopia compared to controls., Methods: Eleven adult amblyopes (26.2 ± 4.4 [SD] years old) and 10 controls (24.6 ± 0.8 years old) with normal or corrected to normal visual acuity (logMAR VA ≤ 0) participated in this study. Using the quick contrast sensitivity function (qCSF) algorithm, we measured balanced CSF which would stimulate the ON and OFF pathways unselectively, and CSFs for increments and decrements that would selectively stimulate the ON and OFF visual pathways. Contrast sensitivity and area under log contrast sensitivity function were extracted for statistical analysis., Results: For the balanced CSF, we found significant interocular differences in sensitivity and area under log contrast sensitivity function in both amblyopes [ F (1,10) = 74.992, p  < 0.001] and controls [ F (1,9) = 35.6, p  < 0.001], while such differences were more pronounced in amblyopes than in controls. For increment and decrement CSFs, we found that the increment sensitivity ( p  = 0.038) and area under log contrast sensitivity function ( p  = 0.001) were significantly lower than the decrement in the amblyopic eye. Such differences between increment and decrement CSFs were not observed in the fellow eye of the amblyopes or in the controls., Conclusion: There is a subtle difference in the contrast sensitivity of the amblyopic eye when exposed to stimulation in the ON and OFF pathways., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Wei, Cheng, Kong, Lin, Hess, Zhou and Reynaud.)

The onset of atherosclerosis (AS) is insidious, and early stage patients have atypical clinical symptoms. After being diagnosed in late stage, it is often prone to sudden and fatal cardiovascular events. Therefore, it is highly desirable to develop precise and efficient diagnosis and therapy strategies of AS. Benefiting from high signal-to-noise ratio, low detection limit, high specificity and sensitivity, a series of activatable fluorescent probes based on atherosclerotic microenvironment have emerged for identification and treatment of AS. In this review, we focus on the atherosclerotic microenvironment and briefly summarize the correlation between the structural transformation and fluorescence signal changes of mono-/double-activatable fluorescent probes upon biomarkers stimulation. Moreover, their cutting-edge progress for AS theranostics is described. Finally, the outlook for activatable theranostic probes based on atherosclerotic microenvironment is discussed to aim at promoting innovative research in imaging-guided precise AS therapy., Competing Interests: The authors declare no competing interests., (© 2024 The Author(s).)

Protein C (PC), a vitamin K-dependent serine protease zymogen in plasma, can be activated by thrombin-thrombomodulin(TM) complex, resulting in the formation of activated protein C (APC). APC functions to downregulate thrombin generation by inactivating active coagulation factors V(FVa) and VIII(FVIIIa). Deficiency in PC increases the risk of venous thromboembolism (VTE). We have identified two unrelated VTE patients with the same heterozygous mutation (c.1384 T > C, p.Ter462GlnextTer17) in PROC. To comprehend the role of this mutation in VTE development, we expressed recombinant PC-Ter462GlnextTer17 in mammalian cells and evaluated its characteristics using established coagulation assay systems. Functional studies revealed a significant impairment in the activation of the mutant by thrombin or thrombin-TM complex. Furthermore, APC-Ter462GlnextTer17 demonstrated diminished hydrolytic activity towards the chromogenic substrate S2366. APTT and FVa degradation assays showed that both the anticoagulant activity of the mutant protein was markedly impaired, regardless of whether protein S was present or absent. These results were further supported by a thrombin generation assay conducted using purified and plasma-based systems. In conclusion, the Ter462GlnextTer17 mutation introduces a novel tail at the C-terminus of PC, leading to impaired activity in both PC zymogen activation and APC's anticoagulant function. This impairment contributes to thrombosis in individuals carrying this heterozygous mutation and represents a genetic risk factor for VTE., Competing Interests: Declaration of competing interest The authors state that there are no conflicts of interest to disclose. This study was supported by Shanghai Pujiang Program (21PJ1409800), Shanghai Natural Science Foundation Program (22ZR1439500), and the General Program of National Natural Science Foundation of China (81870107) to YL; the General Program of National Natural Science Foundation of China (82070137) to XW; the General Program of National Natural Science Foundation of China (81970127) to JD, and the General Program of National Natural Science Foundation of China (82070194) to XC., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Background: Protein C (PC) pathway serves as a major defense mechanism against thrombosis by the activation of PC through the thrombin-thrombomodulin complex and subsequent inactivation of the activated factor (F)V (FVa) and FVIII (FVIIIa) with the assistance of protein S, thereby contributing to hemostatic balance. We identified 2 unrelated patients who suffered from recurrent thrombosis and carried the same heterozygous mutation c.1153A>G, p.Met343Val (M343V), in PROC gene. This mutation had not been previously reported., Objectives: To explore the molecular basis underlying the anticoagulant defect in patients carrying the M343V mutation in PROC., Methods: We expressed PC-M343V variant in mammalian cells and characterized its properties through coagulation assays., Results: Our findings demonstrated that while activation of mutant zymogen by thrombin-thrombomodulin complex was slightly affected, cleavage of chromogenic substrate by APC-M343V was significantly impaired. However, Ca 2+ increased the cleavage efficiency by approximately 50%. Additionally, there was a severe reduction in affinity between APC-M343V and Na + . Furthermore, the inhibitory ability of APC-M343V toward FVa was markedly impaired. Structural and simulation analyses suggested that Val343 might disrupt the potential hydrogen bonds with Trp380 and cause Trp380 to orient closer to His211, potentially interfering with substrate binding and destabilizing the catalytic triad of APC., Conclusion: The M343V mutation in patients adversely affects the reactivity and/or folding of the active site as well as the binding of the physiological substrate to the protease, resulting in impaired protein C anticoagulant activity and ultimately leading to thrombosis., Competing Interests: Declaration of competing interests There are no competing interests to disclose., (Copyright © 2024 International Society on Thrombosis and Haemostasis. Published by Elsevier Inc. All rights reserved.)

Relationships between people in real life are dynamically changed with the interaction process, and due to the heterogeneous preferences, this change is different from person to person. Based on this observation, we propose a new spatial and weighted prisoner's dilemma game model with heterogeneous individuals. Two types of tags, namely, tag-F (concerned about social fairness) and tag-W (concerned about personal well-being), are introduced to describe individuals' different preferences. The link weights indicating the interaction strength between individuals are updated based on different rules that depend on their tags. Through simulations, we verify that a large link weight control factor and a high proportion of tag-F individuals favor the emergence and persistence of cooperation. In addition, an increase in the link weight sensitivity factor favors the evolution of cooperation when the link weight control factor is small. Moreover, while the level of cooperation increases with the proportion of tag-F type in the population, contrary to our intuition, when the population consists entirely of tag-F individuals, in some cases, cooperation cannot reach a higher level compared with the situation when they are mixed with tag-W type. However, at high dilemma intensities, cooperators emerge only when the entire population consists of tag-F type. These results may provide some new insights into the impact of the evolutionary weighted network with heterogeneous preferences on collective cooperative behavior., (© 2024 Author(s). Published under an exclusive license by AIP Publishing.)