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Abstract Electric eel is an excellent example to harness ion-concentration gradients for sustainable power generation. However, current strategies to create electric-eel-inspired power sources commonly involve manual stacking of multiple salinity-gradient power source units, resulting in low efficiency, unstable contact, and poor flexibility. Here we propose a consecutive multimaterial printing strategy to efficiently fabricate biomimetic ionic hydrogel power sources with a maximum stretchability of 137%. The consecutively-printed ionic hydrogel power source filaments showed seamless bonding interface and can maintain stable voltage outputs for 1000 stretching cycles at 100% strain. With arrayed multi-channel printhead, power sources with a maximum voltage of 208 V can be automatically printed and assembled in parallel within 30 min. The as-printed flexible power source filaments can be woven into a wristband to power a digital wristwatch. The presented strategy provides a tool to efficiently produce electric-eel-inspired ionic hydrogel power sources with great stretchability for various flexible power source applications.

Objective: To explore the possible mechanisms by which follicle-stimulating hormone (FSH) regulates postmenopausal osteoporosis through the FSH/FSH receptor (FSHr)/G protein/C/EBPβ/heat shock protein 90 alpha (HSP90α) signalling pathways. Methods: We measured serum FSH, luteinising hormone (LH), and HSP90α levels in the serum and adipose tissue of women of childbearing age and menopausal status. In the in vivo studies, 12 B57CL female mice were divided equally into Sham, OVX, and OVX + FSHr Blocker groups. Serum levels of alkaline phosphatase, FSH, and HSP90α, along with StRACP vitality, were determined, and femur micro-computed tomography was performed. Additionally, FSH, FSHr, G protein, C/EBPβ, and HSP90α levels were assessed using quantitative polymerase chain reaction. Finally, we divided the human multiple myeloma cell line U266 into three groups. The activity of tartrate-resistant acid phosphatase (TRAP) in the supernatant at different stages was detected, and myeloma cells were stained with TRAP. Results: HSP90α levels in adipose tissue supernatant and serum were lower in women of childbearing age than in menopausal women (P

BackgroundCreatinine-cystatin C ratio (CCR) has been demonstrated as an objective marker of sarcopenia in clinical conditions but has not been evaluated as an osteoporosis marker in individuals with normal renal function.MethodsWe selected 271,831 participants with normal renal function from UK Biobank cohort. Multivariable linear/logistic regression and Cox proportional hazards model were used to investigate the phenotypic relationship between CCR and osteoporosis in total subjects and gender-stratified subjects. Based on the genome-wide association study (GWAS) data, linkage disequilibrium regression (LDSC) and Mendelian randomization (MR) analysis were performed to reveal the shared genetic correlations and infer the causal effects, respectively.ResultsAmongst total subjects and gender-stratified subjects, serum CCR was positively associated with eBMD after adjusting for potential risk factors (all P

Background: Pregnant women exhibit an increased demand for nutrients, including vitamins, and a deficiency in vitamins can increase the risk of various pregnancy-related diseases. This study aims to evaluate the vitamin levels in women of different age groups and gestational stages in order to provide targeted dietary guidance and vitamin supplementation strategies. Methods: Pregnant women who registered and attended regular prenatal check-ups at Hangzhou Women’s Hospital from January to December 2021 were selected as study participants. Ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) was used to quantitatively determine the concentrations of vitamins A, D, E, K, B1, B2, B9 (folic acid), and B12 in the serum. Results: The serum vitamin A, B1, and B9 levels decreased with gestational age in the age-matched group, while the vitamin E level increased slightly (p < 0.05). In the advanced-aged group, the levels of vitamins A, B1, B2, and B9 decreased with gestational age, but the levels of vitamins D3, E, K, and B12 slightly increased (p < 0.05). In mid-pregnancy, age-matched women had slightly greater serum levels of vitamins E and K than did women in the advanced-aged group (Z = –2.67, p = 0.008; Z = –2.46, p = 0.014). In late pregnancy, significant differences existed in the serum levels of vitamins B2 and B12 between the two age groups (Z = –2.67, p = 0.008; Z = –2.50, p = 0.013). Conclusions: Vitamin levels varied by gestational stage and age during pregnancy, suggesting that vitamin supplementation should be individualized and stage-adjusted to improve maternal and child health.

BackgroundMultimorbidity has become an important health challenge in the aging population. Accumulated evidence has shown that multimorbidity has complex association patterns, but the further mechanisms underlying the association patterns are largely unknown.MethodsSummary statistics of 14 conditions/diseases were available from the genome-wide association study (GWAS). Linkage disequilibrium score regression analysis (LDSC) was applied to estimate the genetic correlations. Pleiotropic SNPs between two genetically correlated traits were detected using pleiotropic analysis under the composite null hypothesis (PLACO). PLACO-identified SNPs were mapped to genes by Functional Mapping and Annotation of Genome-Wide Association Studies (FUMA), and gene set enrichment analysis and tissue differential expression were performed for the pleiotropic genes. Two-sample Mendelian randomization analyses assessed the bidirectional causality between conditions/diseases.ResultsLDSC analyses revealed the genetic correlations for 20 pairs based on different two-disease combinations of 14 conditions/diseases, and genetic correlations for 10 pairs were significant after Bonferroni adjustment (PConclusionsThis study highlighted the complex mechanisms underlying the association patterns that include the shared genetic components and causal effects among the 14 conditions/diseases. These findings have important implications for guiding the early diagnosis, management, and treatment of comorbidities.

Background: Anemia seriously affects the health and quality of life of the older adult population and may be influenced by various types of environmental metal exposure. Current studies on metals and anemia are mainly limited to single metals, and the association between polymetals and their mixtures and anemia remains unclear. Methods: We determined 11 urinary metal concentrations and hemoglobin levels in 3781 participants. Binary logistic regression and restricted cubic spline (RCS) model were used to estimate the association of individual metals with anemia. We used Bayesian kernel machine regression (BKMR) and Quantile g-computation (Q-g) regression to assess the overall association between metal mixtures and anemia and identify the major contributing elements. Stratified analyses were used to explore the association of different metals with anemia in different populations. Results: In a single-metal model, nine urinary metals significantly associated with anemia. RCS analysis further showed that the association of arsenic (As) and copper (Cu) with anemia was linear, while cobalt, molybdenum, thallium, and zinc were non-linear. The BKMR model revealed a significant positive association between the concentration of metal mixtures and anemia. Combined Q-g regression analysis suggested that metals such as Cu, As, and tellurium (Te) were positively associated with anemia, with Te as the most significant contributor. Stratified analyses showed that the association of different metals with anemia varied among people of different sexes, obesity levels, lifestyle habits, and blood pressure levels. Conclusions: Multiple metals are associated with anemia in the older adult population. A significant positive association was observed between metal mixture concentrations and anemia, with Te being the most important factor. The association between urinary metal concentrations and anemia is more sensitive in the non-hypertensive populations.

Abstract Planar super-oscillatory lens (SOL), a far-field subwavelength-focusing diffractive device, holds great potential for achieving sub-diffraction-limit imaging at multiple wavelengths. However, conventional SOL devices suffer from a numerical-aperture-related intrinsic tradeoff among the depth of focus (DoF), chromatic dispersion and focusing spot size. Here, we apply a multi-objective genetic algorithm (GA) optimization approach to design an apochromatic binary-phase SOL having a prolonged DoF, customized working distance (WD), minimized main-lobe size, and suppressed side-lobe intensity. Experimental implementation demonstrates simultaneous focusing of blue, green and red light beams into an optical needle of ~0.5λ in diameter and DOF > 10λ at WD = 428 μm. By integrating this SOL device with a commercial fluorescence microscope, we perform, for the first time, three-dimensional super-resolution multicolor fluorescence imaging of the “unseen” fine structures of neurons. The present study provides not only a practical route to far-field multicolor super-resolution imaging but also a viable approach for constructing imaging systems avoiding complex sample positioning and unfavorable photobleaching.

After laser welding, the corrosion resistance and microstructure of a newly developed austenitic stainless steel, QN2109, were investigated. A negligible heat-affected zone was found and the segregation of Cr, Mo, Mn, Ni, and N was revealed by EPMA. In contrast to previously reported findings, the corrosion resistance of δ-ferrite obtained here was lower than that of austenite in acidic solutions revealed by SKPFM and MFM, owing to N segregation. The pitting resistance of whole welds was lower than that of base metal (BM), whereas the pitting resistance of the fusion zone was higher than that of BM, owing to the significant decrease in the number of inclusions.

BackgroundTranscutaneous auricular vagus nerve stimulation (taVNS) has emerged as a potentially effective neuromodulation technique for addressing neurological disorders, including disorders of consciousness. Expanding upon our prior clinical study, which demonstrated the superior effectiveness of a 4-week taVNS treatment in patients with minimally conscious state (MCS) compared to those in a vegetative state/unresponsive wakefulness state, the aim of this investigation was to evaluate the safety and therapeutic efficacy of taVNS in individuals with MCS through a sham-controlled randomized double-blind clinical trial.MethodsA cohort of 50 adult patients (male = 33, female = 17) diagnosed with a MCS were randomly assigned to either the active taVNS (N = 25) or sham taVNS (N = 25) groups. The treatment period lasted for 4 weeks, followed by an 8-week follow-up period. The Coma Recovery Scale-Revised (CRS-R) and Glasgow Coma Scale (GCS) were administered at baseline and weekly during the initial 4 weeks. Additionally, the Disability Rating Scale (DRS) was used to assess the patients’ functional abilities via telephone at week 12. Furthermore, various neurophysiological measures, including electroencephalogram (EEG), upper-limb somatosensory evoked potentials (USEP), brainstem auditory evoked potentials (BAEP), and P300 event-related potentials (P300), were employed to monitor changes in brain activity and neural conduction pathways.ResultsThe scores for the active taVNS group in the CRS-R and GCS showed greater improvement over time compared to the sham taVNS group (CRS-R: 1-week, Z = −1.248, p = 0.212; 2-week, Z = −1.090, p = 0.276; 3-week, Z = −2.017, p = 0.044; 4-week, Z = −2.267, p = 0.023. GCS: 1-week, Z = −1.325, p = 0.185; 2-week, Z = −1.245, p = 0.213; 3-week, Z = −1.848, p = 0.065; 4-week, Z = −1.990, p = 0.047). Additionally, the EEG, USEP, BAEP, and P300 also demonstrated significant improvement in the active taVNS group compared to the sham taVNS group at week 4 (EEG, Z = −2.086, p = 0.037; USEP, Z = −2.014, p = 0.044; BAEP, Z = −2.298, p = 0.022; P300 amplitude, Z = −1.974, p = 0.049; P300 latency, t = 2.275, p = 0.027). Subgroup analysis revealed that patients with MCS derived greater benefits from receiving taVNS treatment earlier (CRS-R, Disease duration ≤ 1-month, mean difference = 8.50, 95% CI = [2.22, 14.78], p = 0.027; GCS, Disease duration ≤ 1-month, mean difference = 3.58, 95% CI = [0.14, 7.03], p = 0.044). By week 12, the active taVNS group exhibited lower Disability Rating Scale (DRS) scores compared to the sham taVNS group (Z = −2.105, p = 0.035), indicating a more favorable prognosis for MCS patients who underwent taVNS. Furthermore, no significant adverse events related to taVNS were observed during treatment.ConclusionThe findings of this study suggest that taVNS may serve as a potentially effective and safe intervention for facilitating the restoration of consciousness in individuals diagnosed with MCS. This therapeutic approach appears to enhance cerebral functioning and optimize neural conduction pathways.Clinical trial registrationhttp://www.chictr.org.cn, Identifier ChiCTR2200066629.

Cigars are developing rapidly around the world, but the content characteristics of aroma precursors and their contribution to sensory perception have not been fully elucidated. In this study, 69 aroma precursors from 61 tobaccos of different parts and origins were systematically determined, and the sensory characteristics of middle leaves from different origins and their correlation with aroma precursors were evaluated. The results showed that tobacco parts mainly affected amino acid content, and contents of nicotine, oxalic acid, malic acid, isovaleric acid, cystine, glutarnine, glycine, isoleucine, glutamicacid, asparticacid, and fructose-proline were significantly changed. Tobacco origins mainly influenced the contents of amino acids, polyacids and high fatty acids, and sugar alcohols, and significantly affected the contents of myosmine, anabasine, nonanoic acid, propanetriol, mannitol, mannose, glucose, alanine, arginine, glutarnine, glutamicacid, histidine, serine, threonine, tryptophan, fructose-alanine, and fructose-asparagine. The flavor characteristics were prominent by wood aroma, and the style and quality characteristics varied greatly among different origins of middle leaves. There were 34, 21, and 22 aroma precursors with high correlations with flavor, style, and quality characteristics. This study provides support for regulating the content and coordination of aroma precursors in different tobacco parts and origins to improve sensory characteristics.

Introduction Chronic low back pain (cLBP) is one of the largest and most frequent public health problems worldwide. Tuina is a physical therapy commonly used in China to treat musculoskeletal diseases. Compared with traction, there is little high-quality scientific evidence that can demonstrate the effectiveness of Tuina in the treatment of patients with cLBP. Therefore, the purpose of this clinical trial is to evaluate the effect of massage on cLBP patients compared with traction.Methods and analyses This is a single-centre, assessor-blinded and analyst-blinded prospective randomised controlled trial with two parallel arms. Ninety-four patients with cLBP will be recruited. Three treatments were given every week for a total of 4 weeks. In the Traction group, participants were given traction therapy in the Tuina group, participants will receive a four-step physiotherapy including kneading, rolling, plucking and oblique pulling. The outcomes will be measured at baseline, at the end of treatment, as well as 1 and 2 months after treatment. The primary outcome will be the Hamilton Anxiety Scale after 12 sessions of treatment. The secondary outcomes will be the Visual Analogue Scale, the medical outcomes study Short Form 36, Serum Quantitative Index and genetic testing after 12 sessions of treatment.Ethics and dissemination The study was approved by the Ethics Committee of Yueyang Hospital of Integrated Traditional Chinese and Western Medicine affiliated with Shanghai University of Traditional Chinese Medicine.Trial registration number ChiCTR2200065448.

Abstract Key message The non-linear seemingly unrelated regression mixed-effects model (NSURMEM) and generalized additive model (GAM) were applied for the first time in crown width (CW) additive models of larch (Larix gmelinii Rupr.), birch (Betula platyphylla Suk.), and poplar (Populus davidiana Dode). The crown radii in four directions (CR) exhibited different growth trends and responded differently to tree size and competition variables. In the absence of calibration, GAM was more accurate than NSURMEM for CR and CW predictions. Context Crown radii in four directions (CR) and crown width (CW) are fundamental indicators used to describe tree crowns. The complexity of the CR growth in four directions of different tree species in natural forests is often ignored. There is logical additivity among CR and CW that is also often overlooked. Furthermore, the existing methods applied to CW additive models have some drawbacks. Aims We aim to: (i) evaluate the utility of two new methods in developing CW additive models for larch (Larix gmelinii Rupr.), birch (Betula platyphylla Suk.), and poplar (Populus davidiana Dode) in natural secondary forests of Northeastern China; and (ii) explore the growth patterns of CR in four directions to gain important ecological insights. Methods The non-linear seemingly unrelated regression mixed-effects model (NSURMEM) and generalized additive model (GAM) were used to develop CW additive models and to explore crown growth patterns. The predictive ability of the additive models was evaluated using leave-one-plot-out cross-validation (LOOCV). Results At a fair level without calibration, GAM provided slightly better results than NSURMEM. The response of the four CR to tree size and competition variables is different and may be non-uniform due to complex stand conditions and tree growth strategies. Conclusion The newly provided methods applied to additive models are available for external datasets. GAM is recommended in the absence of calibration. This study has important implications for the understanding of natural forest dynamics and decision-making for critical stand management.

Abstract Hydraulic fracturing technology has played a pivotal role as a key technology in shale gas development, but the geological environmental problems caused have been worrying the public. This paper proposes an advanced step rectangular pulse hydraulic fracturing technique, which can achieve higher pressure and more complex fractures by pumping lower energy fluid. The results indicate that the pressure propagation of step rectangular pulse hydraulic fracturing can be divided into stable and unstable propagation stages. Compared with conventional hydraulic fracturing, longer unsteady propagation time, higher pressure and more complex pressure distribution can be obtained. Step pulse parameters are discussed. We found that the pulse pressure increases linearly with the increase in pulse amplitude. There are optimal pulse frequencies and comprehensive damping coefficients to get optimal fluid pressure in different shale reservoirs. These research results will help reduce geological environmental hazards, and can also provide guidance for the optimization of step rectangular pulse hydraulic fracturing technology in shale reservoirs.

Discovering tactical knowledge aims to extract tactical data derived from battlefield signal data, which is vital in information warfare. The learning and reasoning from battlefield signal information can help commanders make effective decisions. However, traditional methods are limited in capturing sequential and global representation due to their reliance on prior knowledge or feature engineering. The current models based on deep learning focus on extracting implicit behavioral characteristics from combat process data, overlooking the embedded martial knowledge within the recognition of combat intentions. In this work, we fill the above challenge by proposing a dual fusion pipeline introducing graph representation learning into sequence learning to construct tactical behavior sequence graphs expressing implicit martial knowledge, named TBGCN. Specifically, the TBGCN utilizes graph representation learning to represent prior knowledge by building a graph to induce deep learning paradigms, and sequence learning finds the hidden representation from the target’s serialized data. Then, we employ a fusion module to merge two such representations. The significance of integrating graphs with deep learning lies in using the artificial experience of implicit graph structure guiding adaptive learning, which can improve representation ability and model generalization. Extensive experimental results demonstrate that the proposed TBGCN can effectively discover tactical knowledge and significantly outperform the traditional and deep learning methods.

The rotator cuff tear has emerged as a significant global health concern. However, existing therapies fail to fully restore the intricate bone-to-tendon gradients, resulting in compromised biomechanical functionalities of the reconstructed enthesis tissues. Herein, a tri-layered core–shell microfibrous scaffold with layer-specific growth factors (GFs) release is developed using coaxial electrohydrodynamic (EHD) printing for in situ cell recruitment and differentiation to facilitate gradient enthesis tissue repair. Stromal cell-derived factor-1 (SDF-1) is loaded in the shell, while basic fibroblast GF, transforming GF-beta, and bone morphogenetic protein-2 are loaded in the core of the EHD-printed microfibrous scaffolds in a layer-specific manner. Correspondingly, the tri-layered microfibrous scaffolds have a core–shell fiber size of (25.7 ± 5.1) μ m, with a pore size sequentially increasing from (81.5 ± 4.6) μ m to (173.3 ± 6.9) μ m, and to (388.9 ± 6.9 μ m) for the tenogenic, chondrogenic, and osteogenic instructive layers. A rapid release of embedded GFs is observed within the first 2 d, followed by a faster release of SDF-1 and a slightly slower release of differentiation GFs for approximately four weeks. The coaxial EHD-printed microfibrous scaffolds significantly promote stem cell recruitment and direct their differentiation toward tenocyte, chondrocyte, and osteocyte phenotypes in vitro . When implanted in vivo , the tri-layered core–shell microfibrous scaffolds rapidly restored the biomechanical functions and promoted enthesis tissue regeneration with native-like bone-to-tendon gradients. Our findings suggest that the microfibrous scaffolds with layer-specific GFs release may offer a promising clinical solution for enthesis regeneration.

Background: Environmental metal exposure is associated with elevated triglycerides (TG) and the development of chronic kidney disease (CKD). However, the relationship between metal exposure and glomerular filtration rate (GFR) remains uncertain, and the mediating effect of TG between the two is unclear. Methods: This study measured the concentrations of 14 metals in urine samples from 3752 elderly people in the community. The most relevant metals were screened by least absolute shrinkage and selection operator (LASSO) regression. The relationship between combined exposure to multiple metals and abnormal estimated glomerular filtration rate (eGFR) was explored using multivariate logistic regression analysis and Bayesian kernel machine regression (BKMR) analysis. Generalized linear regression models and the Karlson-Holm-Breen (KHB) method were used to assess the mediating effects of TG. Results: In the single-metal model, calcium (Ca), iron (Fe), selenium (Se), strontium (Sr), and thallium (Tl) showed significant negative correlations with the prevalence of abnormal eGFR (all P

The relationship between evolution in grain boundary characteristics and intergranular corrosion properties of cold rolled 443 ultra-pure ferritic stainless steel was investigated. The results showed that low-angle (LA) grain boundaries, special grain boundaries (SGB), and high-angle (HA) grain boundaries had a high intergranular corrosion sensitivity after cold deformation in reducing acid medium. In oxidative acid medium, the LA and SGB boundaries had a high intergranular corrosion resistance in comparison to HA boundaries, and their inhibitory effect on the corrosion continuance of the dynamic recrystallization high-angle (RHA) grain boundaries can improve the corrosion resistance of the RHA grain boundaries.

Spontaneous imbibition behavior of fracturing fluid retained in reservoirs is considered to be one of the important mechanisms of spontaneous oil–water displacement in tight reservoirs. However, the mechanism of imbibition displacement in oil-wet rocks remains unclear. Inspired by the principle of the Marangoni effect, a mathematical model of oil displacement driven by an interfacial tension gradient in an oil-wet capillary tube was established, and the effects of osmotic pressure and hydraulic mechanical dispersion were coupled in the model. The results show that the key component of this mechanism is the breakdown of the initial capillary equilibrium state due to the diffusion of solute, which changes the pressure distribution in the capillary and drives the water to drive oil out of the pores. This can explain the spontaneous imbibition of low-salinity fracturing fluid into the oil-wet tight sample and displace oil without any external force, despite its low permeability. In addition, the related factors were analyzed, including initial concentration, contact angle, solute type, osmotic behavior, and size of the capillary. The results show that for an oil wet reservoir with high salinity, the connate water interfacial tension gradient plays an important role in oil displacement, and the coupling effect of osmotic pressure and hydromechanical dispersion considerably improves recovery. This can provide new insight into the mechanism of spontaneous imbibition of fracturing fluid.

Crevice corrosion behaviors of duplex stainless steel 2205 after hydrogen charging are investigated. The absorbed hydrogen is introduced into the specimens through cathodic galvanostatic method. It has been found the absorbed hydrogen decreases the critical crevice corrosion conditions (breakdown potential, temperature, and chloride ions content), the stable propagation potential, and the repassivation potential. The results reveal that hydrogen can promote the initiation and propagation stages and inhibit the repassivation process of crevice corrosion. Microstructure observation and element distribution analysis find that, owing to the promotion of hydrogen on the precipitation of insoluble molybdenum oxides and molybdate at the bottom of corrosion positions, crevice corrosion of the hydrogen charged specimens exhibits wider but shallower morphologies compared with the uncharged specimens.

IntroductionMonkeypox is a zoonotic disease caused by brick-shaped enveloped monkeypox (Mpox) virus that belongs to the family of ancient viruses known as Poxviridae. Subsequently, the viruses have been reported in various countries. The virus is transmitted by respiratory droplets, skin lesions, and infected body fluids. The infected patients experience fluid-filled blisters, maculopapular rash, myalgia, and fever. Due to the lack of effective drugs or vaccines, there is a need to identify the most potent and effective drugs to reduce the spread of monkeypox. The current study aimed to use computational methods to quickly identify potentially effective drugs against the Mpox virus.MethodsIn our study, the Mpox protein thymidylate kinase (A48R) was targeted because it is a unique drug target. We screened a library of 9000 FDA-approved compounds of the DrugBank database by using various in silico approaches, such as molecular docking and molecular dynamic (MD) simulation.ResultsBased on docking score and interaction analysis, compounds DB12380, DB13276, DB13276, DB11740, DB14675, DB11978, DB08526, DB06573, DB15796, DB08223, DB11736, DB16250, and DB16335 were predicted as the most potent. To examine the dynamic behavior and stability of the docked complexes, three compounds—DB16335, DB15796, and DB16250 —along with the Apo state were simulated for 300ns. The results revealed that compound DB16335 revealed the best docking score (-9.57 kcal/mol) against the Mpox protein thymidylate kinase.DiscussionAdditionally, during the 300 ns MD simulation period, thymidylate kinase DB16335 showed great stability. Further, in vitro and in vivo study is recommended for the final predicted compounds.

BackgroundWhile osteoimmunology interactions between the immune and skeletal systems are known to play an important role in osteoblast development, differentiation and bone metabolism related disease like osteoporosis, such interactions in either bone microenvironment or peripheral circulation in vivo at the single-cell resolution have not yet been characterized.MethodsWe explored the osteoimmunology communications between immune cells and osteoblastic lineage cells (OBCs) by performing CellphoneDB and CellChat analyses with single-cell RNA sequencing (scRNA-seq) data from human femoral head. We also explored the osteoimmunology effects of immune cells in peripheral circulation on skeletal phenotypes. We used a scRNA-seq dataset of peripheral blood monocytes (PBMs) to perform deconvolution analysis. Then weighted gene co-expression network analysis (WGCNA) was used to identify monocyte subtype-specific subnetworks. We next used cell-specific network (CSN) and the least absolute shrinkage and selection operator (LASSO) to analyze the correlation of a gene subnetwork identified by WGCNA with bone mineral density (BMD).ResultsWe constructed immune cell and OBC communication networks and further identified L-R genes, such as JAG1 and NOTCH1/2, with ossification related functions. We also found a Mono4 related subnetwork that may relate to BMD variation in both older males and postmenopausal female subjects.ConclusionsThis is the first study to identify numerous ligand-receptor pairs that likely mediate signals between immune cells and osteoblastic lineage cells. This establishes a foundation to reveal advanced and in-depth osteoimmunology interactions to better understand the relationship between local bone microenvironment and immune cells in peripheral blood and the impact on bone phenotypes.

The new coronavirus SARS-COV-2, which emerged in late 2019 from Wuhan city of China was regarded as causing agent of the COVID-19 pandemic. The primary protease which is also known by various synonymous i.e., main protease, 3-Chymotrypsin-like protease (3CLPRO) has a vital role in the replication of the virus, which can be used as a potential drug target. The current study aimed to identify novel phytochemical therapeutics for 3CLPRO by machine learning-based virtual screening. A total of 4,000 phytochemicals were collected from deep literature surveys and various other sources. The 2D structures of these phytochemicals were retrieved from the PubChem database, and with the use of a molecular operating environment, 2D descriptors were calculated. Machine learning-based virtual screening was performed to predict the active phytochemicals against the SARS-CoV-2 3CLPRO. Random forest achieved 98% accuracy on the train and test set among the different machine learning algorithms. Random forest model was used to screen 4,000 phytochemicals which leads to the identification of 26 inhibitors against the 3CLPRO. These hits were then docked into the active site of 3CLPRO. Based on docking scores and protein-ligand interactions, MD simulations have been performed using 100 ns for the top 5 novel inhibitors, ivermectin, and the APO state of 3CLPRO. The post-dynamic analysis i.e,. Root means square deviation (RMSD), Root mean square fluctuation analysis (RMSF), and MM-GBSA analysis reveal that our newly identified phytochemicals form significant interactions in the binding pocket of 3CLPRO and form stable complexes, indicating that these phytochemicals could be used as potential antagonists for SARS-COV-2.

In the present study, the effects of the seasons and technique on the microbial community and metabolites of traditional bran Cuqu were investigated by the Illumina MiSeq platform and chromatography technology, respectively. These results showed that both significantly impacted the community diversities and metabolic constituents. The abundances of the fungal and bacterial community in Cuqu manufactured in spring (L1) and summer (L2) were higher than those in the other two seasons, i.e., autumn (L3) and winter (L4). Besides, the fungal community structure of spring-Cuqu was similar to that of summer-Cuqu, so was the volatile profile displayed. Of them, Aspergillus in the L type of Cuqu was dominant and its abundance in L1 Cuqu was 70.74%. Moreover, the liquefying and saccharifying activity of L1 Cuqu were the highest among these samples. The liquefying activity, saccharifying activity, and total organic acid content of L1 Cuqu were significantly higher than those of Q Cuqu. Besides, there were significant differences in microbial community structures between them. The enzymes related to the biosynthesis of phenethyl alcohol in Q Cuqu showed a higher expression level than those in L1. The results of this study might provide a theoretical foundation for the optimization of the production process and scientific management of Cuqu.

IVF: Mechanism and correction of increased cancer risk Increased production of ribosomes, the multi-molecular structures inside cells where protein synthesis occurs, is implicated in the enhanced risk of cancer in offspring conceived by in vitro fertilization (IVF), suggesting an approach to reduce this risk. In studies on IVF mouse embryos, Zhiling Li and colleagues at First Affiliated Hospital of Shantou University Medical College in China found that harmful effects of natural oxidizing agents induced overproduction of specific protein components of ribosomes. This increased the embryos’ susceptibility to tumorigenesis through a particular molecular signaling pathway. Exposure to the natural antioxidant chemical epigallocatechin-3-gallate (EGCG) corrected the overproduction and protected the embryos. EGCG, found in green tea, is being widely investigated for its anticancer effects. Exposing embryos to EGCG during IVF might protect the resulting children from increased risk of tumor formation in later life.

Chemical analysis of atmospheric aerosols by conventional analytical methods is usually required to perform complicated and time-consuming sample preparation processes. In recent decades, ambient ionization mass spectrometry (AI-MS) methods have been proven to be simple, rapid, and effective analytical tools for direct analysis of various complex samples. In this work, we applied porous paper filters for direct adsorptive sampling of tobacco smoke, and then the sampled paper filters were performed the emitters of the paper spray ionization (PSI) device. An auto-sampling device was made to control the generation and collection of tobacco smoke. Nicotine, the typical compound of tobacco smoke, was used to optimize the key conditions of auto-sampling. Moreover, different types of tobacco smoke were also compared with multivariate variable analysis, and the makers of tobacco smoke from different sources of tobacco smoke were investigated. By using this method, direct sampling and analysis of a single tobacco sample can be completed within minutes. Overall, our results show that PSI-MS is a powerful tool that integrates collection, extraction, ionization, and identification analytes in smoke.

BackgroundEffective identification of high-risk rheumatoid arthritis (RA) individuals is still a challenge. Whether the combined effects of multiple previously reported genetic loci together with lifestyle factors can improve the prediction of RA risk remains unclear.MethodsBased on previously reported results and a large-scale Biobank dataset, we constructed a polygenic risk score (PRS) for RA to evaluate the combined effects of the previously identified genetic loci in both case-control and prospective cohorts. We then evaluated the relationships between several lifestyles and RA risk and determined healthy lifestyles. Then, the joint effects of healthy lifestyles and genetic risk on RA risk were evaluated.ResultsWe found a positive association between PRS and RA risk (OR = 1.407, 95% confidence interval (CI) = 1.354~1.463; HR = 1.316, 95% CI = 1.257~1.377). Compared with the low genetic risk group, the group with intermediate or high genetic risk had a higher risk (OR = 1.347, 95% CI = 1.213~1.496; HR = 1.246, 95% CI = 1.108~1.400) (OR = 2.169, 95% CI = 1.946~2.417; HR = 1.762, 95% CI = 1.557~1.995). After adjusting for covariates, we found protective effects of three lifestyles (no current smoking, regular physical activity, and moderate body mass index) on RA risk and defined them as healthy lifestyles. Compared with the individuals with low genetic risks and favorable lifestyles, those with high genetic risks and unfavorable lifestyles had as high as OR of 4.637 (95%CI = 3.767~5.708) and HR of 3.532 (95%CI = 2.799~4.458).ConclusionsIn conclusion, the integration of PRS and lifestyles can improve the prediction of RA risk. High RA risk can be alleviated by adopting healthy lifestyles but aggravated by adopting unfavorable lifestyles.

With the rapid increase of text information on the Internet, intelligent question answering has attracted considerable attention in many different domains. Many studies use intelligent question answering as an answer selection task and use local or global features to encode sentence representation, which has been proven to be very effective. However, due to the different characteristics of the corpora in different domains or languages, these methods usually suffer a great reduction in performance and cannot produce a sentence representation with good qualities. In this paper, we introduce Hybrid Model combining Local Composite features and Global features into a Siamese network (HM-LCGS) to alleviate the issue. The framework consists of a novel convolution neural network like architecture called local composite features convolution neural network to extract sufficient semantic information of the text from different granularity, shortcut connection to combine local composite features into pre-trained embeddings, alignment layer to mine the correlation between question and answer and bidirectional Long Short-Term Memory to encode the final sentence representation. The experimental results on InsuranceQA and cMedQA datasets show that with suitable granularity selection and embedding method, our proposed model can achieve competitive performance compared with other state-of-the-art models.

Hydrogen peroxide is one of the most important reactive oxygen species, which plays a vital role in many physiological and pathological processes. A dramatic increase in H2O2 levels is a prominent feature of cancer. Therefore, rapid and sensitive detection of H2O2 in vivo is quite conducive to an early cancer diagnosis. On the other hand, the therapeutic potential of estrogen receptor beta (ERβ) has been implicated in many diseases including prostate cancer, and this target has attracted intensive attention recently. In this work, we report the development of the first H2O2-triggered ERβ-targeted near-infrared fluorescence (NIR) probe and its application in imaging of prostate cancer both in vitro and in vivo. The probe showed good ERβ selective binding affinity, excellent H2O2 responsiveness and near infrared imaging potential. Moreover, in vivo and ex vivo imaging studies indicated that the probe could selectively bind to DU-145 prostate cancer cells and rapidly visualizes H2O2 in DU-145 xenograft tumors. Mechanistic studies such as high-resolution mass spectrometry (HRMS) and density functional theory (DFT) calculations indicated that the borate ester group is vital for the H2O2 response turn-on fluorescence of the probe. Therefore, this probe might be a promising imaging tool for monitoring the H2O2 levels and early diagnosis studies in prostate cancer research.

Currently, we are facing an increasing trend in the use of unmanned aerial vehicles (UAV) in various activities, both civilian and military. The application of UAVs in the battlefield has received extensive attention amid a global new military revolution. A UAV cluster is a large and complex real-time feedback system that integrates a communication and sensor network, control system, calculation, and physical process. The heterogeneous UAVs conduct complex behaviors, which requires a comprehensive description and analysis of UAV cluster modeling. The integrated modeling of the heterogeneous UAV cluster is of great significance and value to test and verify the new combat mode. In this paper, we present a novel representation framework based on the Petri nets. We used a spatial-temporal hybrid Petri net to illustrate the discrete state and continuous process of a heterogeneous UAV cluster system, and effectively achieved the fusion of a physical and computational process and interaction event modeling in the cluster system. Finally, the formal verification of UAV cluster attack mission modeling was carried out by UPPAAL, suggesting the proposed modeling method is feasible and effective.

Ornithobacterium rhinotracheale (ORT) has been associated with avian respiratory disease. On coinfection with other pathogens, ORT can cause serious health problems in avian species, leading to financial losses. To monitor the serologic prevalence of ORT in chicken flocks in China, 1,280 sera were collected to determine ORT antibodies among 64 flocks from 15 provinces of China using a commercial ELISA kit. The overall seroprevalence of ORT among the birds tested was 44.06%. In younger chickens, the serum positive rate was lower than that in older chickens, and with increased age, the serum positive rates increased. Older chickens had not only higher positive rates but also higher antibody levels. These data indicated that ORT infections were common in China. Because an ORT vaccine is currently not available, good disease management and biosecurity measures are required for effective disease control.

To study the differences in phenolic compounds between tobacco smokers’ saliva and mainstream smoke, a method was developed for the analysis of 12 phenolic compounds in saliva and mainstream smoke based on ultrahigh-performance liquid chromatography with fluorescence detection (UPLC-FLD). The contents and distributions of phenolic compounds in tobacco smokers’ saliva and mainstream smoke were compared. The results were as follows: (1) Phenolic compounds were quantitatively analyzed by the internal standard method using 4-fluorophenol as an internal standard. For smokers’ saliva samples, the limits of quantification (LOQs) ranged from 2.2 to 19.1 μg/L, and the recoveries were from 80.2% to 119.2% at the three spiked levels. For mainstream smoke samples, the LOQs ranged from 0.03 to 0.26 μg/cig, and the recoveries ranged from 84.9% to 107.0% at the three spiked levels. (2) The contents of phenolic compounds from 14 cigarettes in mainstream smoke and smokers’ saliva were determined. In mainstream smoking, the main phenolic compounds were hydroquinone, catechol, phenol, meta- and para-Cresol, and o-methylhydroquinone. In smokers’ saliva, the main phenolic compounds were phenol and meta- and para-Cresol and the contents of phenolic compounds in smokers’ saliva from different cigarettes were significantly different. (3) The content distribution patterns of phenolic compounds in smokers’ saliva differed from those in mainstream smoke. The predominant phenolic compound in mainstream smoke was dihydroxybenzene, while monophenols predominated in smokers’ saliva. (4) The contents of phenolic compounds from five kinds of cigarettes were analyzed in the saliva of different smokers using principal component analysis, which indicated that cigarettes with different sensory effects were clearly distinguished by differences in the contents of phenolic compounds in saliva.

The shale fracture permeability is critical in determining gas production and deep CO2 sequestration performance. Moreover, how shale fracture permeability evolves after interactions with supercritical carbon dioxide (Sc-CO2) should be understood to constrain the shale reservoir permeability and evaluate the long-term sealing ability of shale formations. In this research, we conducted soaking experiments with shale fractures and Sc-CO2 at various times and then measured the shale fracture permeability and hydraulic aperture evolution under different stress states. Additionally, we quantify the chemical compositions, pore characteristics, fracture surface roughness alternation through X-ray diffraction, nuclear magnetic resonance, scanning electron microscope, and optical profilometry techniques. Our results indicate that soaking with Sc-CO2 will dramatically increase the shale fracture permeability and aperture due to the calcite and dolomite dissolution. This free-face dissolution process will remove the mineral particles in the fracture surface, resulting in larger pores, peaks, and valleys in the fracture surfaces. This process may last for seven days, and after that, chemical reactions may terminate, and the rock-Sc-CO2 system turns stable. Our results explain how Sc-CO2 alters the shale fracture permeability through the chemical dissolution of specific minerals from a microscale analysis.

With the increase in environmental pressure and rapid development of renewable energy technologies, an integrated energy system has been recognized as an effective approach to accommodate large-scale renewables and achieve environmental sustainability. While an integrated energy system significantly improves energy efficiency, the interaction between different energy systems may also bring multiple operational risks to its reliability, which necessitates an effective reliability assessment technique. In this paper, we proposed a multi-level fuzzy evaluation model based on combined empowerment for the reliability evaluation of an integrated energy system. The analytic hierarchy process method and entropy weight method were used to calculate the weight of each index in the evaluation model. Fuzzy evaluation matrix was constructed by the membership degree of a single factor, which was defined by the fuzzy comprehensive evaluation method. The multi-level fuzzy evaluation results were obtained based on single-level evaluation results. Finally, case studies were carried out based on a practical integrated energy system; we proposed 5 first-level indicators such as reliability and economy and 12 second-level indicators such as mean time of incapacity. The simulation results (85.15) showed the effectiveness and advantages of the proposed model.

In this paper, a large-scale experimental system was established to identify the migration and distribution laws of complex fracture proppant in shale reservoir volume fracturing. With this system, the effects of secondary fracture angle, fluid displacement, proppant concentration and size, fracturing fluid viscosity and other factors on the migration and distribution of proppant were tested, and the migration and distribution of proppant in primary/secondary fractures were analyzed. The following results were obtained. First, the fluid flow pattern in fractures transforms gradually from laminar flow into turbulent flow with the increase of fracture supporting height. Second, the migration modes of proppant in fractures mainly include suspended migration and gliding migration. Third, the distribution form of proppant in primary fractures before branching is related to secondary fracture angle, fluid displacement and proppant concentration and size, among which the fluid displacement is the most important factor. Fourth, the mass ratio of proppant in primary fractures after branching is proportional to the secondary fracture angle, fluid displacement, fracturing fluid viscosity and proppant concentration and size, and is inversely proportional to the flow ratio between secondary fractures and primary fractures. Fifth, the mass ratio of proppant in secondary fractures after branching is proportional to fluid displacement, fracturing fluid viscosity and flow ratio between secondary fractures and primary fractures, and is inversely proportional to secondary fracture angle and proppant concentration and size. Sixth, the angle at the leading edge of proppant bank in the primary fractures after branching is proportional to the proppant concentration and size and the flow ratio between secondary fractures and primary fractures, but is inversely proportional to secondary fracture angle, fluid displacement and fracturing fluid viscosity. Seventh, the angle at the leading edge of proppant bank in the secondary fractures after branching is proportional to the secondary fracture angle and the proppant concentration and size, but is inversely proportional to the fluid displacement, fracturing fluid viscosity and flow ratio between secondary fractures and primary fractures. In conclusion, the research results can provide a theoretical support for proppant optimization and program design of shale reservoir volume fracturing. Keywords: Shale, Reservoir, Volume fracturing, Complex fracture, Proppant, Migration and distribution, Experimental system

Gonadotropins play vital roles in the regulation of female reproductive ability and fertility. Our study aimed to determine the effects of superovulation induced by increasing doses of equine chorionic gonadotropin [eCG; also referred to as pregnant mare serum gonadotropin (PMSG)] on the developmental competence of mouse embryos and on aneuploidy formation during in vitro fertilization (IVF). eCG dose-dependently enhanced the oocyte yield from each mouse. Administration of 15 IU eCG significantly reduced the fertilization rate and the formation of four-cell embryos and blastocysts and increased the risk of chromosome aneuploidy. The IVF-derived blastocysts in the 15 IU eCG treatment group had the fewest total cells, inner cell mass (ICM) cells and trophectoderm (TE) cells. Moreover, more blastocysts and fewer apoptotic cells were observed in the 0, 5, and 10 IU eCG treatment groups than in the 15 IU eCG treatment group. We also investigated reactive oxygen species (ROS) levels and variations in several variables: mitochondrial membrane potential (MMP); active mitochondria; mitochondrial superoxide production; adenosine triphosphate (ATP) content; spindle structures; chromosome karyotypes; microfilament distribution; and the expression of Aurora B [an important component of the chromosomal passenger complex (CPC)], the spindle assembly checkpoint (SAC) protein mitotic arrest deficient 2 like 1 (MAD2L1), and the DNA damage response (DDR) protein γH2AX. Injection of 15 IU eCG increased ROS levels, rapidly reduced MMP, increased active mitochondria numbers and mitochondrial superoxide production, reduced ATP content, increased abnormal spindle formation rates, and induced abnormalities in chromosome number and microfilament distribution, suggesting that a high dose of eCG might alter developmental competence and exert negative effects on IVF-obtained mouse embryos. Additionally, the appearance of γH2AX and the significantly increased expression of Aurora B and MAD2L1 suggested that administration of relatively high doses of eCG caused Aurora B-mediated SAC activation triggered by ROS-induced DNA damage in early mouse IVF-derived embryos for self-correction of aneuploidy formation. These findings improve our understanding of the application of gonadotropins and provide a theoretical basis for gonadotropin treatment.

Schistosomiasis is among the major neglected tropical diseases and effective prevention by boosting the immune system is still not available. T cells are key cellular components governing adaptive immune response to various infections. While common laboratory mice, such as C57BL/6, are highly susceptible to schistosomiasis, the SD rats are extremely resistant. However, whether adaptive immunity is necessary for such natural resistance to schistosomiasis in rats remains to be determined. Therefore, it is necessary to establish genetic model deficient in T cells and adaptive immunity on the resistant SD background, and to characterize liver pathology during schistosomiasis. In this study we compared experimental schistosomiasis in highly susceptible C57BL/6 (B6) mice and in resistant SD rats, using cercariae of Schistosoma japonicum. We observed a marked T cell expansion in the spleen of infected B6 mice, but not resistant SD rats. Interestingly, CD3e-/- B6 mice in which T cells are completely absent, the infectious burden of adult worms was significantly higher than that in WT mice, suggesting an anti-parasitic role for T cells in B6 mice during schistosome infection. In further experiments, we established Lck deficient SD rats by using CRISPR/Cas9 in which T cell development was completely abolished. Strikingly, we found that such Lck deficiency in SD rats severely impaired their natural resistance to schistosome infection, and fostered parasite growth. Together with an additional genetic model deficient in T cells, the CD3e-/- SD rats, we confirmed the absence of T cell resulted in loss of natural resistance to schistosome infection, but also mitigated liver immunopathology. Our further experiments showed that regulatory T cell differentiation in infected SD rats was significantly decreased during schistosomiasis, in contrast to significant increase of regulatory T cells in infected B6 mice. These data suggest that T cell mediated immune tolerance facilitates persistent infection in mice but not in SD rats. The demonstration of an important role for T cells in natural resistance of SD rats to schistosomiasis provides experimental evidences supporting the rationale to boost T cell responses in humans to prevent and treat schistosomiasis.

Background: Cervical cancer is a common gynecologic malignancy worldwide, mainly developing in women aged about 50 years old. Currently, oxaliplatin (L-OHP) was widely used as a first-line chemotherapeutic drug to treat various tumors, including cervical cancer. The emergence of L-OHP resistance during chemotherapy has largely limited the clinical efficacy of chemotherapy. Numerous studies have demonstrated that abnormal expression of microRNAs (miRNAs) was associated with tumorigenesis and the development of cancer drug resistance. Methods: miR-34a-5p and Murine Double Minute 4 (MDM4) in cervical cancer cells was detected via RT-qPCR and Western blot assay. Cell proliferation and apoptosis were observed via ov-MDM4 and si-MDM4, Cell counting kit (CCK)-8 and flow cytometry analysis after transfection with miR-34a-5p inhibitor, miR-34a-5p mimics, respectively. Dual-luciferase reporter assay was utilized to confirm the associativity between miR-34a-5p and MDM4. Results: miR-34a-5p was significantly down-regulated whereas MDM4 was increased in cervical cancer tumor tissues and cells. Compared with Hela cells, miR-34a-5p was further decreased and MDM4 was elevated in Hela/L-OHP cells. miR-34a-5p significantly inhibited Hela/L-OHP cell viability and promoted apoptosis. Similar to the effects of miR-34a-5p, MDM4 knockdown inhibited Hela/L-OHP cell proliferation, but induced apoptosis. miR-34a-5p directly targeted MDM4 and could improve sensitivity of cervical cancer cells to L-OHP chemosensitivity by targeting MDM4 expression in vitro. Conclusions: miR-34a-5p can improve sensitivity of cervical cancer cells to L-OHP chemosensitivity, serving as a potential curative target for cervical cancer chemotherapy.

Objective To identify serum protein markers in midtrimester that predict preterm delivery. Methods A retrospective case–control study randomly selected patients that experienced spontaneous preterm birth and healthy control patients that experienced a normal delivery at term. A proteomic analysis was undertaken using the data-independent acquisition method. Results A total of 30 singleton pregnant women were randomly selected from 12 800 pregnant women: 15 women had a spontaneous preterm birth (group Y) and 15 age- and body mass index-matched women gave birth at term (group D). All of the patients provided serum at 15–20 weeks of gestation. A total of 39 differentially expressed proteins were identified. Compared with group D, 24 proteins were upregulated and 15 were downregulated in the preterm group Y. Using Kyoto Encyclopedia of Genes and Genomes pathway enrichment, the 24 upregulated proteins were significantly enriched in the complement and coagulation cascade pathways. Search Tool for the Retrieval of Interacting Genes Furthermore (STRING) analysis showed that apolipoprotein A-II (apoA-II) and alpha-2-antiplasmin (α2-AP), two upregulated proteins, were key nodes in the STRING protein–protein network. Conclusions These findings suggest that apoA-II and α2-AP might be new markers for predicting preterm delivery in the midtrimester.

In zygotes, the capacity of G2/M checkpoint and DNA repair mechanisms to respond to DNA damage varies depending on different external stressors. In our previous studies, we found that mild oxidative stress induced a G2/M phase delay in mouse zygotes fertilized in vitro, due to the activation of the spindle assembly checkpoint. However, it is unclear whether the G2/M phase delay involves G2 arrest, triggered by activation of the G2/M checkpoint, and whether AMPK, a highly conserved cellular energy sensor, is involved in G2 arrest and DNA damage repair in mouse zygotes. Here, we found that mouse zygotes treated with 0.03 mM H2O2 at 7 h post-insemination (G1 phase), went into G2 arrest in the first cleavage. Furthermore, phosphorylated H2AX, a specific DNA damage and repair marker, can be detected since the early S phase. We also observed that oxidative stress induced phosphorylation and activation of AMPK. Oxidative stress-activated AMPK first localized in the cytoplasm of the mouse zygotes in the late G1 phase and then translocated to the nucleus from the early S phase. Overall, most of the activated AMPK accumulated in the nuclei of mouse zygotes arrested in the G2 phase. Inhibition of AMPK activity with Compound C and SBI-0206965 abolished oxidative stress-induced G2 arrest, increased the activity of CDK1, and decreased the induction of cell cycle regulatory proteins p53 and p21. Moreover, bypassing G2 arrest after AMPK inhibition aggravated oxidative stress-induced DNA damage at M phase, increased the apoptotic rate of blastocysts, and reduced the formation rate of 4-cell embryos and blastocysts. Our results suggest the G2/M checkpoint and DNA repair mechanisms are operative in coping with mild oxidative stress-induced DNA damage. Further, AMPK activation plays a vital role in the regulation of the oxidative stress-induced G2 arrest through the inhibition of CDK1 activity via p53/p21 pathways, thereby facilitating the repair of DNA damage and the development and survival of oxidative stress-damaged embryos. Our study provides insights into the molecular mechanisms underlying oxidative-stress induced embryonic developmental arrest, which is crucial for the development of novel strategies to ensure viable embryo generation.

BackgroundGenome-wide association studies (GWASs) routinely identify loci associated with risk factors for osteoporosis. However, GWASs with relatively small sample sizes still lack sufficient power to ascertain the majority of genetic variants with small to modest effect size, which may together truly influence the phenotype. The loci identified only account for a small percentage of the heritability of osteoporosis. This study aims to identify novel genetic loci associated with DXA-derived femoral neck (FNK) bone mineral density (BMD) and quantitative ultrasound of the heel calcaneus estimated BMD (eBMD), and to detect shared/causal variants for the two traits, to assess whether the SNPs or putative causal SNPs associated with eBMD were also associated with FNK-BMD.MethodsNovel loci associated with eBMD and FNK-BMD were identified by the genetic pleiotropic conditional false discovery rate (cFDR) method. Shared putative causal variants between eBMD and FNK-BMD and putative causal SNPs for each trait were identified by the colocalization method. Mendelian randomization analysis addresses the causal relationship between eBMD/FNK-BMD and fracture.ResultsWe identified 9,500 (cFDR < 9.8E-6), 137 (cFDR < 8.9E-4) and 124 SNPs associated with eBMD, FNK-BMD, and both eBMD and FNK-BMD, respectively, with 37 genomic regions where there was a SNP that influences both eBMD and FNK-BMD. Most genomic regions only contained putative causal SNPs associated with eBMD and 3 regions contained two distinct putative causal SNPs influenced both traits, respectively. We demonstrated a causal effect of FNK-BMD/eBMD on fracture.ConclusionMost of SNPs or putative causal SNPs associated with FNK-BMD were also associated with eBMD. However, most of SNPs or putative causal SNPs associated with eBMD were not associated with FNK-BMD. The novel variants we identified may help to account for the additional proportion of variance of each trait and advance our understanding of the genetic mechanisms underlying osteoporotic fracture.

The tumor-suppressor gene tumor protein p53 (TP53) is one of the most commonly mutated genes in human lung cancer, and TP53 mutations are associated with a worsened prognosis and causes resistance to cancer therapy. RNA sequencing and TP53 mutation data were downloaded to determine specific TP53-associated signature based on differentially expressed genes between patients with lung squamous cell carcinoma (LUSC) with wild type (TP53 WT) and mutated (TP53MUT) TP53. We investigated the predictive value of this signature on the immune microenvironment, tumor mutational burden (TMB), and likelihood of response to immunotherapy and chemotherapy. In total, 1,556 differentially expressed genes were identified based on TP53 mutation status. Three genes (KLK6, MUC22 and CSN1S1) identified by univariate and multivariate Cox regression analyses, comprised the prognostic signature which was an independent and specific prognostic marker of overall survival in patients with LUSC. A nomogram was also established to validate this signature for clinical use. Moreover, the high-risk group was characterized by increased infiltration of monocytes and macrophages M1, and decreased T cells CD8 and T cells follicular helper. High-risk group exhibited a higher TMB, and was much more sensitive to immunotherapy and chemotherapy. KLK6 and CSN1S1 expression and the prognostic prediction values were further validated in clinical samples. The derived TP53-associated signature is a specific and independent prognostic biomarker for LUSC patients, and could provide potential prognostic biomarker or therapeutic targets for the development of novel immunotherapies and chemotherapies.

The fracture propagation in hydraulic fracturing is described as a nonlinear problem dynamic boundary. Due to the limitation of mesh refinement, it is difficult to obtain the real crack propagation path using conventional numerical methods. Meshless methods (MMs) are an effective method to eliminate the dependence on the computational grid in the simulation of fracture propagation. In this paper, a hydraulic fracture propagation model is established based on the element-free Galerkin (EFG) method by introducing jump and branch enrichment functions. Based on the proposed method, three types of fracturing technology are investigated. The results reveal that the stress interference between fractures has an important impact on the propagation path. For the codirectional fracturing simultaneously, fractures propagate in a repel direction. However, the new fracture is attracted and eventually trapped by the adjacent fracture in the sequential fracturing case. For the opposite simultaneous fracturing in multiwells, two fractures with a certain lateral spacing will deflect toward each other. The effect of stress shadow should be used rationally in the optimization of construction parameters; for the single well multistage fracturing, the stage spacing should be out of stress inversion area, while for the simultaneous fracturing of multiple wells, stress inversion zones should be used to maximize communication between natural fractures. Overall, this study establishes a novel and effective approach of using MM to simulate the propagation of hydraulic fractures, which can serve as a useful reference for understanding the mechanism of hydraulic fracture propagation under various conditions.

Abstract Background The apicomplexan parasite Babesia orientalis, the causative agent of water buffalo babesiosis in China, is widespread in central and south China, resulting in a huge economic loss annually. Currently, there is no effective vaccine or drug against this disease. Babesia bovis and Plasmodium falciparum were reported to possess an apicoplast which contains the methylerythritol phosphate (MEP) pathway inhibitable by fosmidomycin, suggesting that the pathway could serve as a drug target for screening new drugs. However, it remains unknown in B. orientalis. Methods Primers were designed according to the seven MEP pathway genes of Babesia microti and Babesia bovis. The genes were cloned, sequenced and analyzed. The open reading frames (ORFs) of the first two enzyme genes, 1-deoxy-D-xylulose 5-phosphate synthase (BoDXS) and 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (BoDXR), were cloned into the pET-32a expression vector and expressed as a Trx-tag fusion protein. Rabbit anti-rBoDXS and rabbit anti-rBoDXR antibodies were generated. Western blot was performed to identify the native proteins of BoDXS and BoDXR in B. orientalis. Fosmidomycin and geranylgeraniol were used for inhibition assay and rescue assay, respectively, in the in vitro cultivation of B. orientalis. Results The seven enzyme genes of the B. orientalis MEP pathway (DXS, DXR, IspD, IspE, IspF, IspG and IspH) were cloned and sequenced, with a full length of 2094, 1554, 1344, 1521, 654, 1932 and 1056 bp, respectively. BoDXS and BoDXR were expressed as Trx-tag fusion proteins, with a size of 95 and 67 kDa, respectively. Western blot identified a 77 kDa band for the native BoDXS and a 49 kDa band for the native BoDXR. The drug assay results showed that fosmidomycin could inhibit the growth of B. orientalis, and geranylgeraniol could reverse the effect of fosmidomycin. Conclusions Babesia orientalis has the isoprenoid biosynthesis pathway, which could be a potential drug target for controlling and curing babesiosis. Considering the high price and instability of fosmidomycin, further studies should focus on the screening of stable and cheap drugs.

Abstract Background The spherical body, a membrane bound organelle localized in the apical organelle complex, is unique to Babesia and Theileria spp. The spherical body proteins (SBPs) secreted by spherical bodies include SBP1, SBP2, SBP3 and SBP4. Up to now, only SBP3 has been characterized in Babesia orientalis. Methods The BoSBP4 gene was amplified from cDNA and gDNA and cloned into the pGEX-6P-1 vector by homologous recombination, sequenced and analyzed by bioinformatics tools. The amino acid (aa) sequence of BoSBP4 was compared with that of Babesia bovis and Babesia bigemina as well as SBP3 of B. orientalis. The immunoreactivity was evaluated by incubating recombinant BoSBP4 (rBoSBP4) with the serum of B. orientalis-infected water buffalo. The native form of BoSBP4 was identified by incubating lysate of B. orientalis-infected water buffalo erythrocytes with the anti-rBoSBP4 mouse serum. The cellular localization of BoSBP4 was determined by indirect immunofluorescence assay. Results The full length of the BoSBP4 gene was estimated to be 945 bp without introns, encoding a 314 aa polypeptide with a predicted molecular weight of 37 kDa. The truncated recombinant protein was expressed from 70 to 945 bp as a GST fusion protein with a practical molecular weight of 70 kDa. BoSBP4 shared a 40% and 30% identity with B. bovis and B. bigemina, respectively. Furthermore, it was 31% identical to SBP3 of B. orientalis. BoSBP4 was identified in the lysate of B. orientalis-infected water buffalo erythrocytes with a molecular weight of 37 kDa, corresponding to the expected molecular mass of BoSBP4. The result of rBoSBP4 with positive serum revealed that BoSBP4 can elicit an immune response to B. orientalis-infected water buffalo. The cellular localization of BoSBP4 was detected to be adjacent to the merozoite nucleus in the intracellular phase, followed by the diffusion of the fluorescence of BoSBP4 into the cytoplasm of B. orientalis-infected erythrocytes as puncta-like specks and a gradual increase of the fluorescence. Conclusions In this study, SBP4 in B. orientalis was characterized for the first time. It may play a key role in interaction with the host cell by being secreted into the cytoplasm of the B. orientalis-infected erythrocytes to facilitate parasite growth and reproduction.

Abstract: Sweet spots in the shale reservoirs of the Lower Silurian Longmaxi Formation in Weiyuan 201 Block of Sichuan Basin were predicted quantitatively using seismic data and fuzzy optimization method. First, based on seismic and rock physics analysis, the rock physics characteristics of the reservoirs were determined, and elastic parameters sensitive to shale reservoirs with high gas content were selected. Second, data volumes with high precision of the elastic parameters were obtained from pre-stack simultaneous inversion. The horizontal distribution of key parameters for shale gas evaluation were calculated based on the results of rock physics analysis. Then, the fuzzy evaluation equation was established by fuzzy optimization method with test and logging data of horizontal wells with similar operation conditions. key parameters affecting the productivity of horizontal wells were sorted out and the weights of them in the sweet spots quantitative prediction were worked out by fuzzy optimization to set up a sweet spots evaluation system. Three classes of shale gas reservoirs which including two kinds of sweet spots were predicted with the above procedure, and the sweet spots have been predicted quantitatively by combining the above prediction results with the testing production. The testing results of 7 verification wells proved the reliability of the prediction results. Key words: Sichuan Basin, Lower Silurian Longmaxi Formation, shale gas, sweet spots, quantitative prediction, fuzzy optimisation

Abstract Background The spherical body is a distinct organelle only existing in Babesia and Theileria. Spherical body proteins (SBPs) are secreted from spherical bodies and incorporated into the cytoplasm of infected erythrocytes during invasion and post-invasion stages. Four different SBP homologues (SBP1, SBP2, SBP3 and SBP4) have been identified in Babesia bovis and Babesia bigemina. So far, there has been no report available about the identification of SBPs in Babesia orientalis. Methods The SBP3-like in B. orientalis (BoSBP3-like) was cloned, sequenced, characterized and compared to the SBP3 sequences of B. bovis and B. bigemina by bioinformatics analyses. The BoSBP3-like gene was truncated into three fragments: BoSBP3-like-1 (915 bp), BoSBP3-like-2 (1311 bp) and BoSBP3-like-3 (1011 bp), which were amplified and cloned into the expression vector pET-28a and expressed as three truncated recombinant (His-fusion) proteins. The immunogenicity, native forms and localization of BoSBP3-like were identified by western blot and indirect immunofluorescence assay (IFA). Results The BoSBP3-like gene was intronless with an open reading frame (ORF) of 3237 bp, encoded a 1079 amino acid polypeptide with a predicted size of 135 kDa, and contained a cysteine-rich region, three dispersing FAINT domains and a signal peptide (1–16 aa) at the N-terminus. The amino acid sequence of BoSBP3-like was 61.6 and 35.0% identical to that of B. bovis and B. bigemina, respectively. BoSBP3-like was identified as 135 kDa in the parasite lysate by rabbit antiserum against the truncated recombinant BoSBP3-like-1 (rBoSBP3-like-1). Three specific bands corresponding to rBoSBP3-like-1 (1–305 aa, 43 kDa), rBoSBP3-like-2 (306–742 aa, 58 kDa) and rBoSBP3-like-3 (743–1079 aa, 52 kDa) were detected by reaction with serum from B. orientalis-infected buffalo. The BoSBP3-like was not only localized in the spherical body of B. orientalis but also in the cytoplasm of infected erythrocytes of buffalo as puncta-like protein specks at both single and paired parasite development stages. Conclusions Through secretion into the cytoplasm of infected erythrocytes, BoSBP3-like may play a significant role in adaptation, interaction, and modification related to the host environment to benefit the growth and survival of Babesia. BoSBP3-like could react with the serum from B. orientalis-infected buffalo, but not healthy buffalo, implicating that BoSBP3-like is highly antigenic and may serve as a candidate diagnostic antigen for the detection of B. orientalis.

Abstract RIZ1 has been studied as a tumor suppressor and may play a role in metabolic diseases related to the Western style diet, such as cancer and obesity. The Akt pathway is known to play a role in both cancer and obesity, and a link between Akt and RIZ1 has also been found. To better understand the role of RIZ1 in obesity and cancer, we investigated how RIZ1 regulates the expression of Akt3. We found that overexpression of RIZ1 in HEK293 cells reduced the expression of Akt3 protein. Luciferase reporter activity of Akt3 gene promoter was significantly reduced in cells co-transfected with RIZ1. Recombinant proteins of RIZ1 was able to bind the Akt3 promoter in vitro, and chromatin immunoprecipitation assay also demonstrated the ability of RIZ1 binding to the Akt3 promoter in vivo. Overexpression of RIZ1 increased H3K9 methylation on the Akt3 promoter. These results identify Akt3 as a target of RIZ1 regulation and expand our understanding of the Akt pathway in cancer and obesity.

Abstract Background Schistosomiasis in the People’s Republic of China (PRC) can be traced back to antiquity. In the past 60 years, the Chinese government has made great efforts to control this persistent disease with elimination slated by 2020 through the implementation of a comprehensive control strategy. This strategy aims to reduce the role of bovines and humans as sources of infection as a pre-requisite for elimination through transmission interruption. The goal of elimination will be achievable only by the implementation of a sustainable surveillance and control system, with sensitive diagnosis a key feature so that the true disease burden is not underestimated. Currently used diagnostics lack the necessary sensitivity to accurately determine the prevalence of Schistosoma japonicum infection in areas with low infection intensities. It is of critical importance to find and treat people and to identify animals with low-level infections if the National Control Programme for China is to achieve schistosomiasis elimination. Methods We evaluated a real-time polymerase chain reaction (qPCR) assay using 633 human stool samples collected from five villages in Hunan, Anhui, Hubei, and Jiangxi provinces, and 182 bovine (70 cattle and 112 buffalo) stool samples obtained from four villages in Hunan, Anhui, and Jiangxi provinces in the PRC. All stool samples were subjected to the miracidium hatching test (MHT, a diagnostic procedure used in the National Schistosomiasis Control Programme) and the qPCR assay. Samples positive by MHT were subjected to either the Kato-Katz technique for humans, or the formalin-ethyl acetate sedimentation-digestion (FEA-SD) procedure for bovines, to determine infection intensities. Results The qPCR assay exhibited a high level of sensitivity in the detection of S. japonicum infections. With both the human and bovine samples, a significantly higher prevalence was determined using the qPCR assay (11.06% humans, 24.73% bovines) than with the MHT (0.93% humans, 7.69% bovines). The animal contamination index (calculated using data obtained with the qPCR technique) for all positive bovines was 27 618 000 eggs per day, indicating a considerable amount of environmental egg contamination that would be underestimated using less sensitive diagnostic procedures. Conclusions The qPCR assay we have evaluated will be applicable as a future field diagnostic and surveillance tool in low-transmission zones where schistosomiasis elimination is targeted and for monitoring post-intervention areas to verify that elimination has been maintained.