Mechanistic insights into poly(C)-binding protein hnRNP K resolving i-motif DNA secondary structures.

I-motifs are four-strand noncanonical secondary structures formed by cytosine (C)-rich sequences in living cells. The structural dynamics of i-motifs play essential roles in many cellular processes, such as telomerase inhibition, DNA replication, and transcriptional regulation. In cells, the structural dynamics of the i-motif can be modulated by the interaction of poly(C)-binding proteins (PCBPs), and the interaction is closely related to human health, through modulating the transcription of oncogenes and telomere stability. Therefore, the mechanisms of how PCBPs interact with i-motif structures are fundamentally important. However, the underlying mechanisms remain elusive. I-motif structures in the promoter of the c-MYC oncogene can be unfolded by heterogeneous nuclear ribonucleoprotein K (hnRNP K), a PCBP, to activate its transcription. Here, we selected this system as an example to comprehensively study the unfolding mechanisms. We found that the promoter sequence containing 5 C-runs preferred folding into type-1245 to type-1234 i-motif structures based on their folding stability, which was further confirmed by single-molecule FRET. In addition, we first revealed that the c-MYC i-motif structure was discretely resolved by hnRNP K through two intermediate states, which were assigned to the opposite hairpin and neighboring hairpin, as further confirmed by site mutations. Furthermore, we found all three KH (hnRNP K homology) domains of hnRNP K could unfold the c-MYC i-motif structure, and KH2 and KH3 were more active than KH1. In conclusion, this study may deepen our understanding of the interactions between i-motifs and PCBPs and may be helpful for drug development.

[Knowledge graph analysis of pyroptosis research in traditional Chinese medicine based on VOSviewer and CiteSpace].

To explore the research hotspots and frontier directions of pyroptosis in the field of traditional Chinese medicine(TCM), the authors searched CNKI and Web of Science for literature related to pyroptosis in TCM, screened literature according to the search strategy and inclusion criteria, and analyzed the publication trend of the included literature. VOSviewer was used to draw author cooperation and keyword co-occurrence network diagrams, and CiteSpace was employed for keyword clustering, emergence, and timeline view. Finally, 507 Chinese literature and 464 English literature were included, and it was found that the number of Chinese and English literature was increasing rapidly year by year. The co-occurrence of the authors showed that in terms of Chinese literature, there was a representative research team composed of DU Guan-hua, WANG Shou-bao and FANG Lian-hua, and for English literature, the representative research team was composed of XIAO Xiao-he, BAI Zhao-fang and XU Guang. The network visualization of Chinese and English keywords revealed that inflammation, apoptosis, oxidative stress, autophagy, organ damage, fibrosis, atherosclerosis, and ischemia-reperfusion injury were the primary research diseases and pathological processes in TCM; berberine, resveratrol, puerarin, na-ringenin, astragaloside Ⅳ, and baicalin were the representative active ingredients; NLRP3/caspase-1/GSDMD, TLR4/NF-κB/NLRP3, and p38/MAPK signaling pathways were the main research pathways. Keyword clustering, emergence, and timeline analysis indicated that the pyroptosis research in TCM focused on the mechanism of TCM monomers and compounds intervening in diseases and pathological processes. Pyroptosis is a research hotspot in the area of TCM, and the current discussion mainly focuses on the mechanism of the therapeutic effect of TCM.

[Robot-assisted radical prostatectomy: An update].

The incidence of prostate cancer (PCa) is on a gradual rise. For localized PCa, radical prostatectomy is a main treatment option among many others, and laparoscopic radical prostatectomy is even considered as a gold treatment standard. However, with the development of robots and improvement of clinicians' surgical experience and understanding of prostatic anatomy, robot-assisted radical prostatectomy has been gaining a wide application. This review focuses on the development, advantages and disadvantages of robot-assisted radical prostatectomy with a view to providing some reference for clinicians.

[Evidence mapping of clinical research on 20 Chinese patent medicines for hypertension].

In this study, evidence mapping was employed to sort out and summarise the evidence from clinical studies of Chinese patent medicines for hypertension and to understand the evidence distribution in related studies. Chinese patent medicines for hypertension were searched from Medicine Catalogue for National Basic Medical Insurance, Employment Injury Insurance, and Maternity Insu-rance(2021) and Chinese Pharmacopoeia(2020). Relevant articles(published from January 1, 2016 to February 14, 2022) were retrieved from Cochrane Library, PubMed, Web of Science, CNKI, Wanfang, VIP, and SinoMed. Then, the evidence distribution was analysed based on description, tables, and bubble charts. A total of 31 Chinese patent medicines were identified and 20 were finally included, involving 111 articles. The basic information of the 20 Chinese patent medicines, the number of related articles, the hypertension staging and traditional Chinese medicine(TCM) syndrome types of the subjects, sample size, interventions, and outcome indicators were compared. The results showed Chinese patent medicines with the function of pacifying liver and eliminating wind were frequently studied, and most of them were single-center, small-sample, short-period randomized controlled trials. They failed to highlight the key and advantages of TCM. A wide variety of outcome indicators were involved, and in addition to blood pressure, surrogate outcome indicators and composite outcome indicators were emphasized. However, health economic indicators, quality of life, and damage to target organs such as blood vessels and heart, were rarely used.

Genome-wide characterization and expression profiling of B3 superfamily during ethylene-induced flowering in pineapple (Ananas comosus L.).

BACKGROUND: The B3 superfamily (B3s) represents a class of large plant-specific transcription factors, which play diverse roles in plant growth and development process including flowering induction. However, identification and functional surveys of B3 superfamily have not been reported in ethylene-induced pineapple flowering (Ananas comosus). RESULTS: 57 B3 genes containing B3 domain were identified and phylogenetically classified into five subfamilies. Chromosomal localization analysis revealed that 54 of 57 AcB3s were located on 21 Linkage Groups (LG). Collinearity analysis demonstrated that the segmental duplication was the main event in the evolution of B3 gene superfamily, and most of them were under purifying selection. The analysis of cis-element composition suggested that most of these genes may have function in response to abscisic acid, ethylene, MeJA, light, and abiotic stress. qRT-PCR analysis of 40 AcB3s containing ethylene responsive elements exhibited that the expression levels of 35 genes were up-regulated within 1 d after ethephon treatment and some were highly expressed in flower bud differentiation period in stem apex, such as Aco012003, Aco019552 and Aco014401. CONCLUSION: This study provides a basic information of AcB3s and clues for involvement of some AcB3s in ethylene-induced flowering in pineapple.

Quantification of sphingosine 1-phosphate by validated LC-MS/MS method revealing strong correlation with apolipoprotein M in plasma but not in serum due to platelet activation during blood coagulation.

Sphingosine 1-phosphate (S1P) is a signalling sphingolipid affecting multiple cellular functions of vascular and immune systems. It circulates at submicromolar levels bound to HDL-associated apolipoprotein M (apoM) or to albumin. S1P in blood is mainly produced by platelets and erythrocytes, making blood sampling for S1P quantification delicate. Standardisation of sampling is thereby of great importance to obtain robust data. By optimising and characterising the extraction procedure and the LC-MS/MS analysis, we have developed and validated a highly specific and sensitive method for S1P quantification. Blood was collected from healthy individuals (n = 15) to evaluate the effects of differential blood sampling on S1P levels. To evaluate correlation between S1P and apoM in different types of plasma and serum, apoM was measured by ELISA. The method showed good accuracy and precision in the range of 0.011 to 0.9 µM with less than 0.07 % carryover. We found that the methanol precipitation used to extract S1P co-extracted apoM and several other HDL-proteins from plasma. The platelet-associated S1P was released during coagulation, thus increasing the S1P concentration to double in serum as compared to that in plasma. Gel filtration chromatography revealed that the platelet-released S1P was mainly bound to albumin. This explains why the strong correlation between S1P and apoM levels in plasma is lost upon the clotting process and hence not observed in serum. We have developed, characterised and validated an efficient, highly sensitive and specific method for the quantification of S1P in biological material.

[Monitoring the Redox States of Thioredoxin in Protein-Protein Interaction Using Intrinsic Fluorescence Probe].

The cellular redox states directly affect cell proliferation, differentiation and apoptosis, and the redox states changes is particularly important to the regulation of cell survival or death. Thioredoxin is a kind of oxidation regulatory protein which is widely exists in organisms, and the change of redox states is also an important process in redox regulation. In this work, we have used the site-directed mutagenesis of protein, SDS-polyacrylamide gel electrophoresis fluorescence spectroscopy and circular dichroism etc., to investigate redox states changes between TRX (E. coli) and glutathione peroxidase(GPX3) during their interaction. By observing the fluorescence spectra of TRX and its mutants, we have studied the protein interactions as well as the redox states switching between oxidation state TRX and the reduced state GPX3. The results demonstrate the presence of interactions and electron exchanges occurring between reduced GPX3 and oxidized TRX, which is of significance for revealing the physical and chemical mechanism of TRX in intracellular signal transduction.

Molecular Prevalence, Genetic Diversity, and Tissue Tropism of Bartonella Species in Small Mammals from Yunnan Province, China.

Bartonella is an intracellular parasitic zoonotic pathogen that can infect animals and cause a variety of human diseases. This study investigates Bartonella prevalence in small mammals in Yunnan Province, China, focusing on tissue tropism. A total of 333 small mammals were sampled from thirteen species, three orders, four families, and four genera in Heqing and Gongshan Counties. Conventional PCR and real-time quantitative PCR (qPCR) were utilized for detection and quantification, followed by bioinformatic analysis of obtained DNA sequences. Results show a 31.5% detection rate, varying across species. Notably, Apodemus chevrieri, Eothenomys eleusis, Niviventer fulvescens, Rattus tanezumi, Episoriculus leucops, Anourosorex squamipes, and Ochotona Thibetana exhibited infection rates of 44.4%, 27.7%, 100.0%, 6.3%, 60.0%, 23.5%, and 22.2%, respectively. Genetic analysis identified thirty, ten, and five strains based on ssrA, rpoB, and gltA genes, with nucleotide identities ranging from 92.1% to 100.0%. Bartonella strains were assigned to B. grahamii, B. rochalimae, B. sendai, B. koshimizu, B. phoceensis, B. taylorii, and a new species identified in Episoriculus leucops (GS136). Analysis of the different tissues naturally infected by Bartonella species revealed varied copy numbers across different tissues, with the highest load in spleen tissue. These findings underscore Bartonella's diverse species and host range in Yunnan Province, highlighting the presence of extensive tissue tropism in Bartonella species naturally infecting small mammalian tissues.

[Current status of morning blood pressure control and medication of hypertensive patients in Beijing].

OBJECTIVE: To explore the current status of morning blood pressure and medication of hypertensive patients in Beijing. METHODS: This study included 2187 hypertensive patients who visited the ambulance of our cardiology department in the morning (7:00-10:00) from March 2012 to April 2012. Patients were divided into three groups: no antihypertensive agent group, single antihypertensive drug therapy group (include CCB, ARB, ACEI, ß-blocker) and combined drug therapy group at least one month. Blood pressure control rate was compared among the groups. RESULTS: Target blood pressure was not reached in 1193 patients (54.6%), most patients took CCB and the target blood pressure was not reached in 61.7% (295/478) patients taking CCB. There was no significant difference on target blood pressure uncontrolled rate among the four single drug subgroups (CCB, ARB, ACEI, ß-blocker). The blood pressure uncontrolled rate was 46.3% (63/136) for amlodipine, 70.5% (55/78) for nifedipine and 73.8% (31/42) for felodipine. There OR of uncontrolled blood pressure rate was 0.36 (amlodipine vs. nifedipine, 95%CI:0.20-0.65) and 0.31% (amlodipine vs. felodipine, 95%CI:0.14-0.66). CONCLUSION: The morning blood pressure uncontrolled rate is high in hypertensive patients visiting Beijing tertiary hospitals. Amlodipine is possible superior to nifedipine and felodipine on morning blood pressure control in this patient cohort.

Organic carbon preservation in wetlands: Iron oxide protection vs. thermodynamic limitation.

The sequestration of organic carbon (OC) in wetland sediments is influenced by the presence of oxygen or lack thereof. The mechanisms of OC sequestration under redox fluctuations, particularly by the co-mediation of reactive iron (Fe) protection and thermodynamic limitation by the energetics of the OC itself, remain unclear. Over the past 26 years, a combination of field surveys and remote sensing images had revealed a strong decline in both natural and constructed wetland areas in Tianjin. This decline could be attributed to anthropogenic landfill practices and agricultural reclamation efforts, which may have significant impacts on the oxidation-reduction conditions for sedimentary OC. The Fe-bound OC (CBD extraction) decreased by 2 to 10-fold (from 8.3 to 10% to 0.7-4.5%) with increasing sediment depth at three sites with varying water depths (WD). The high-resolution spectro-microscopy analysis demonstrated that Fe (oxyhydr)oxides were colocalized with sedimentary OC. Corresponding to lower redox potential, the nominal oxidation state of C (NOSC), which corresponds to the energy content in OC, became more negative (energy content increased) with increasing sediment depth. Taken together, the preservation of sedimentary OC is contingent on the prevailing redox conditions: In environments where oxygen availability is high, reactive Fe provides protection for OC, while in anoxic environments, thermodynamic constraints (i.e., energetic constraints) limit the oxidation of OC.

[Association between fractional flow reserve and quantitative coronary angiography parameters in intermediate coronary artery stenosis].

OBJECTIVE: To explore the relationship between quantitative coronary angiography (QCA) parameters and fractional flow reserve (FFR) for identifying ideal angiographic parameters predictive of myocardial ischemia. METHODS: The study included 121 lesions with QCA and FFR data from 106 patients [mean age: (63 ± 10) years]. The lesions were grouped into FFR > 0.75 group and FFR ≤ 0.75 group. Assessed parameters by QCA included percentage diameter stenosis, minimum luminal diameter (MLD), percentage area stenosis, minimum luminal area (MLA), reference vessel diameter (RVD) and lesion length (LL). Correlation analysis was used to identify the relationship between QCA parameters and FFR value, and receiver operating characteristic (ROC) curve was used to determine parameters predictive of FFR ≤ 0.75. RESULTS: LL was significantly higher [(14.8 ± 7.9) mm vs. (10.7 ± 5.4) mm, P = 0.024] while MLD [(1.47 ± 0.31) mm vs. (1.82 ± 0.51) mm, P = 0.028], RVD [(2.30 ± 0.50) mm vs. (2.81 ± 0.64) mm, P = 0.036], and MLA [(2.30 ± 1.50) mm(2) vs. (3.60 ± 2.30) mm(2), P = 0.038] were significantly lower in FFR ≤ 0.75 group than in FFR > 0.75 group. LL (r = -0.209, P = 0.040) was negatively correlated with FFR, and MLD (r = 0.414, P = 0.040), RVD (r = 0.303, P = 0.000) and MLA (r = 0.315, P = 0.002) were positively correlated with FFR. ROC analysis showed that MLD ≥ 1.6 mm was the best cut-off value to predict FFR > 0.75 with sensitivity 63%, specificity 82%, and positive predictive value 96%. CONCLUSIONS: QCA derived anatomic parameters of intermediate coronary lesions correlate to FFR value in some extent. MLD ≥ 1.6 mm is the best cut-off value to predict FFR > 0.75 in patients with intermediate coronary lesions.

Prognostic differences of catestatin among young and elderly patients with acute myocardial infarction.

BACKGROUND: Previous studies have reported inconsistent findings regarding the association between catestatin and outcomes of acute myocardial infarction (AMI). This study aims to investigate the prognostic value of catestatin for long-term outcomes in patients with AMI. METHODS: One hundred and sixty-five patients with AMI were enrolled in this series. The plasma catestatin levels at baseline and clinical data were collected. All patients were followed up for four years to investigate whether there were major adverse cardiovascular events (MACEs), including cardiovascular death, recurrent AMI, rehospitalization for heart failure, and revascularization. RESULTS: There were 24 patients who had MACEs during the follow-up period. The MACEs group had significantly lower plasma catestatin levels (0.74±0.49 ng/mL vs. 1.10±0.79 ng/mL, P=0.033) and were older (59.0±11.4 years old vs. 53.2±12.8 years old, P=0.036). The rate of MACEs was significantly higher in the elderly group (≥60 years old) than in the young group (<60 years old) (23.8% [15/63] vs. 8.8% [9/102], P=0.008). The catestatin level was significantly lower in the MACEs group than that in the non-MACEs group (0.76±0.50 ng/mL vs. 1.31±0.77 ng/mL, P=0.012), and catestatin was significantly associated with MACEs (Kaplan Meier, P=0.007) among the elderly group, but not in the young group (Kaplan Meier, P=0.893). In the Cox proportional hazards regression, high catestatin was one of the independent factors for predicting MACEs after adjustment for other risk factors (hazard ratio 0.19, 95% confidence interval 0.06-0.62, P=0.006) among elderly patients. CONCLUSIONS: Elderly AMI patients with lower plasma catestatin levels are more likely to develop MACEs. Catestatin may be a novel marker for the long-term prognosis of AMI, especially in elderly patients.

Luminescent Metal Complexes for Bioassays in the Near-Infrared (NIR) Region.

Near-infrared (NIR, 700-1700 nm) luminescent imaging is an emerging bioimaging technology with low photon scattering, minimal autofluorescence, deep tissue penetration, and high spatiotemporal resolution that has shown fascinating promise for NIR imaging-guided theranostics. In recent progress, NIR luminescent metal complexes have attracted substantially increased research attention owing to their intrinsic merits, including small size, anti-photobleaching, long lifetime, and metal-centered NIR emission. In the past decade, scientists have contributed to the advancement of NIR metal complexes involving efforts to improve photophysical properties, biocompatibility, specificity, pharmacokinetics, in vivo visualization, and attempts to exploit new ligand platforms. Herein, we summarize recent progress and provide future perspectives for NIR metal complexes, including d-block transition metals and f-block lanthanides (Ln) as NIR optical molecular probes for bioassays.

Symptom Status, Clinical Subtypes, and Prognosis in Patients With Atrial Fibrillation from the Middle East Region (from the Gulf-SAFE Registry).

Atrial fibrillation (AF) is often asymptomatic. The prognosis of asymptomatic AF is at least similar or worse than symptomatic AF, but there are no such data from Middle East patients with AF. The Gulf-SAFE (Gulf Survey of Atrial Fibrillation Events) registry is a multicenter prospective survey of patients presenting with AF to participate medical institutions in 6 countries in the Gulf region. We investigated the prognostic outcomes of patients with asymptomatic AF in relation to clinical subtypes. A total of 2043 patients with AF were included; 541 were identified as having asymptomatic AF (26.5%) who tended to be older, with higher prevalences of hypertension, heart failure, coronary artery disease, diabetes, stroke, renal dysfunction, chronic obstructive pulmonary disease, and had higher Congestive heart failure, Hypertension, Age ≥75, Stroke (2 points), Congestive heart failure, Hypertension, Age ≥75 (2 points), Diabetes, Stroke (2 points), Vascular disease, Age 65-74, Sex category (CHA2DS2-VASc), and Hypertension, Age ≥65, Stroke, Bleeding history, liable INR, Elderly, Drug or alcohol use (HAS-BLED) scores (all p <0.05). After multivariable adjustment, asymptomatic AF was associated with higher risks of stroke/systematic embolism (SE) (adjusted odds ratio [aOR] 2.18, 95% confidence interval [CI] 1.10 to 4.34), all-cause mortality (aOR 2.85, 95% CI 1.90 to 4.28), and the composite outcome of stroke/SE, bleeding, and all-cause mortality (aOR 1.74, 95% CI 1.26 to 2.41). Patients with asymptomatic AF had fewer admissions for AF (aOR 0.53, 95% CI 0.32 to 0.83) and heart failure (aOR 0.58, 95% CI 0.38 to 0.86). The increased risk of stroke/SE in asymptomatic AF was more prominent among paroxysmal AF subtype (p for interaction = 0.028). In conclusion, in the Gulf-SAFE registry, patients with asymptomatic AF represent a nonbenign entity with worse outcomes compared with symptomatic AF. In paroxysmal AF, the higher risks of events were more prominent. The absence of "warning signs" and lack of timely admission in asymptomatic AF may be major reasons for the unfavorable prognosis.

Metabonomics: A Useful Tool to Reveal Underlying Relationships between Altered Chinese Medicine Syndromes and Ultrafiltration in Treatment of Heart Failure.

OBJECTIVE: To reveal the underlying relationships between Chinese medicine (CM) syndromes and ultrafiltration (UF) in the treatment of heart failure based on a metabonomic approach. METHODS: Seventeen acute decompensated heart failure (ADHF) patients were enrolled, and their CM syndromes before and after UF were collected. In addition, their venous plasma collected before and after UF was used for liquid chromatographmass spectrometer-based metabonomic analysis. Both reversed phase liquid chromatography and hydrophilic interaction liquid chromatography were used to analyze the plasma samples. Partial least-squares to latent structure-discriminant analyses were used for data analysis. RESULTS: An obvious difference was observed pre- and post-treatment. A total of 17 potential biomarkers associating with alterd syndromes with UF including hypoxanthine, 1-methylhistidine, phytosphingosine, O-decanoyl-R-carnitine, etc. were screened out, showing a significant change after UF. The major adjusted metabolic pathways were purine metabolism, histidine metabolism, leucine and isoleucine metabolism, arginine and proline metabolism, carnitine shuttle, sphingolipid metabolism and phospholipid metabolism. CONCLUSIONS: Metabonomic approach is a useful tool to identify potential biomarkers of altered syndromes link to UF and could provide a theoretical basis for further research on the therapeutic mechanism of UF combined with CM.

Analysis of risk factors for early stent thrombosis in the Chinese population: A multicenter restrospective study.

BACKGROUND: The predictive scoring systems for early stent thrombosis (EST) remains blank in China. The study aims to evaluate the risk factors and conduct a prediction model of EST in the Chinese population. METHODS: EST was defined as thrombosis that occurs within the first 30 days after primary percutaneous coronary intervention (PCI). Patients from ten Chinese hospitals diagnosed as stent thrombosis (ST) from January 2010 to December 2016 were retrospectively included as the study group. A control group (1 case:2 controls) was created by including patients without ST, major adverse cardiovascular events, or cerebrovascular events during follow-up. The present study evaluated 426 patients with single-vessel lesions and ultimately included 40 patients with EST and 80 control patients, who were included to identify factors that predicted EST and to develop a prediction scoring system. The other 171 patients without integrated 1:2 pair were used for external validation. RESULTS: EST was independently associated with a low hemoglobin concentration (adjusted odds ratio [OR] 0.946, 95% confidence interval [95% CI] 0.901-0.993, P=0.026), a high pre-PCI Synergy between Percutaneous Coronary Intervention with Taxus and Cardiac Surgery (SYNTAX) score (OR 1.166, 95% CI 1.049-1.297, P=0.004), and a DAPT (DAPT) duration of <30 days (OR 28.033, 95% CI 5.302-272.834, P<0.001). The simple EST prediction score provided an area under the curve (AUC) of 0.854 (95% CI 0.777-0.932, P<0.001) with 70.0% sensitivity and 90.0% specificity, and 0.742 (95% CI 0.649-0.835, P<0.001) with 54.5% sensitivity and 81.0% specificity for external validation dataset. CONCLUSIONS: EST may be independently associated with DAPT discontinuation within 30 days, a low hemoglobin concentration, and a high SYNTAX score. The scoring system also has a good ability to predict the risk of EST and may be useful in the clinical setting.

Bayesian Adversarial Spectral Clustering with Unknown Cluster Number.

Spectral clustering is a popular tool in many unsupervised computer vision and machine learning tasks. Recently, due to the encouraging performance of deep neural networks, many conventional spectral clustering methods have been extended to the deep framework. Although these deep spectral clustering methods are quite powerful and effective, learning the cluster number from data is still a challenge. In this paper, we aim to tackle this problem by integrating the spectral clustering, generative adversarial network and low rank model within a unified Bayesian framework. First, we adapt the low rank method to the cluster number estimation problem. Then, an adversarial-learning-based deep clustering method is proposed and incorporated. When introducing the spectral clustering method into our model clustering procedure, a hidden space structure preservation term is proposed. Via a Bayesian framework, the structure preservation term is embedded into the generative process, which can then be used to deduce a spectral clustering in the optimization procedure. Finally, we derive a variational-inference-based method and embed it into the network optimization and learning procedure. Experiments on different datasets prove that our model has the cluster number estimation capability and show that our method can outperform many similar graph clustering methods.

Single-molecule probing the duplex and G4 unwinding patterns of a RecD family helicase.

RecD family helicases play an important role in prokaryotic genome stability and serve as the structural models for studying superfamily 1B (SF1B) helicases. However, RecD-catalyzed duplex DNA unwinding behavior and the underlying mechanism are still elusive. RecD family helicases share a common proto-helicase with eukaryotic Pif1 family helicases, which are well known for their outstanding G-quadruplex (G4) unwinding ability. However, there are still controversial points as to whether and how RecD helicases unfold G4 structures. Here, single-molecule fluorescence resonance energy transfer (smFRET) and magnetic tweezers (MT) were used to study Deinococcus radiodurans RecD2 (DrRecD2)-mediated duplex DNA unwinding and resolution of G4 structures. A symmetric, repetitive unwinding phenomenon was observed on duplex DNA, revealed from the strand switch and translocation of one monomer. Furthermore, we found that DrRecD2 was able to unwind both parallel and antiparallel G4 structures without obvious topological preferences. Surprisingly, the unwinding properties of RecD on duplex and G4 DNA are different from those of Pif1. The findings provide an example, in which the patterns of two molecules derived from a common ancestor deviate during evolution, and they are of significance for understanding the unwinding mechanism and function of SF1B helicases.

[Increased postprocedural neutrophil count is an independent predictive factor of poor prognosis in acute ST elevation myocardial infarction patients treated with percutaneous coronary intervention].

OBJECTIVE: The aim of the study was to evaluate the prognostic value of the postprocedural neutrophil count in patients with first acute ST elevation myocardial infarction (STEMI) treated with successful primary percutaneous coronary intervention (PCI). METHODS: A total of 226 consecutive STEMI patients underwent successful primary PCI were enrolled. Electrocardiograms were recorded before PCI and 2 hours after PCI. Neutrophil counts were measured within 12 hours after PCI. All patients were followed up for 2 years. Logistic regression analysis was used to evaluate predictive values of postprocedural neutrophil for ST-segment resolution (STR) after PCI and for death, non-fatal myocardial infarction and heart failure at 30 days and 2 years post PCI. Time-to-event analyses were performed using the Kaplan-Meier survival curves in patients with various ranges of postprocedural neutrophil counts. RESULTS: Postprocedural neutrophil count ranged from 2.83x10(9)/L to 18.74x10(9)/L, first quartile, median and fourth quartile were 5.66x10(9)/L, 7.38x10(9)/L and 9.34x10(9)/L respectively. Multivariable logistic analysis showed that when postprocedural neutrophil count increased 1x10(9)/L, the risk of non-STR increased 2.28 fold (OR: 2.28, P=0.009), the risk of death (OR: 1.63, P=0.010) and heart failure (OR: 1.16, P=0.035) at 30 days increased 1.63 and 1.16 folds respectively, and the risk of death (OR: 1.29, P=0.003) and heart failure (OR: 1.20, P=0.007) at 2 years increased 1.29 and 1.20 folds respectively, but the risk of non-fatal myocardial infarction was not affected by postprocedural neutrophil count. Furthermore, the patients with postprocedural neutrophil count>or=9.34x10(9)/L had significant lower 30-day (89.1% vs. 99.1% vs. 98.2%, P=0.010) and 2-year (82.4% vs. 96.1% vs. 96.3%, P=0.003) survival rates compared with the patients with postprocedural neutrophil count from 5.66x10(9)/L to 9.33x10(9)/L and the patients with postprocedural neutrophil count<5.66x10(9)/L (all P<0.05). CONCLUSION: Postprocedural neutrophil count is an independent predictor of short- and long-term death and heart failure in first acute STEMI patients treated with successful primary PCI.

New functional test for the TFPIα cofactor activity of Protein S working in synergy with FV-Short.

Essentials Protein S and FV-Short are synergistic cofactors to Tissue Factor Pathway Inhibitor α (TFPIα). An assay for the TFPIα synergistic cofactor activity of protein S with FV-Short was developed. The assay was specific for the synergistic TFPIα-cofactor activity of free protein S. Protein S deficient individuals with known mutations were correctly distinguished from controls. SUMMARY: Background Protein S is an anticoagulant cofactor to both activated protein C and tissue factor pathway inhibitor (TFPIα). The TFPIα-cofactor activity of protein S is stimulated by a short isoform of factor V (FV-Short), the two proteins functioning in synergy. Objective Using the synergistic TFPIα-cofactor activity between protein S and FV-Short to develop a functional test for plasma protein S. Patients/Methods TFPIα-mediated inhibition of FXa in the presence of FV-Short, protein S and negatively charged phospholipid vesicles was monitored in time by synthetic substrate S2765. TFPIα, FXa and FV-Short were purified proteins, whereas diluted plasma from protein S deficient patients or controls were used as source for protein S. Results The assay was specific for free protein S demonstrating good correlation to free protein S plasma levels (r = 0.92) with a Y-axis intercept of -5%. Correlation to concentrations of total protein S (free and C4BPß+-bound) was lower (r = 0.88) and the Y-axis intercept was +46%, which is consistent with the specificity for free protein S. The test distinguished protein S-deficient individuals from 6 families with known ProS1 mutations from family members having no mutation. Protein S levels of warfarin-treated protein S deficient cases were lower than protein S in cases treated with warfarin for other causes. Conclusions We describe a new assay measuring the TFPIα-cofactor activity of plasma protein S. The test identifies type I/III protein S deficiencies and will be a useful tool to detect type II protein S deficiency having defective TFPIα-cofactor activity.