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Abstract Bruton tyrosine kinase inhibitor (BTKi) combined with rituximab-based chemotherapy benefits diffuse large B-cell lymphoma (DLBCL) patients. However, drug resistance is the major cause of relapse and death of DLBCL. In this study, we conducted a comprehensive analysis BTKi-resistance related genes (BRRGs) and established a 10-gene (CARD16, TRIP13, PSRC1, CASP1, PLBD1, CARD6, CAPG, CACNA1A, CDH15, and NDUFA4) signature for early identifying high-risk DLBCL patients. The resistance scores based on the BRRGs signature were associated with prognosis. Furthermore, we developed a nomogram incorporating the BRRGs signature, which demonstrated excellent performance in predicting the prognosis of DLBCL patients. Notably, tumor immune microenvironment, biological pathways, and chemotherapy sensitivity were different between high- and low-resistance score groups. Additionally, we identified TRIP13 as a key gene in our model. TRIP13 was found to be overexpressed in DLBCL and BTKi-resistant DLBCL cell lines, knocking down TRIP13 suppresses cell proliferation, promotes cell apoptosis, and enhances the apoptosis effect of BTKi on DLBCL cells by regulating the Wnt/β-catenin pathway. In conclusion, our study presents a novel BRRGs signature that could serve as a promising prognostic marker in DLBCL, and TRIP13 might be a potential therapeutic target for resistant DLBCL.

ABSTRACT: Aims: Electroacupuncture (EA) at Shuigou (GV26) and Baihui (GV20) has shown therapeutic benefits for stroke patients. Given that natural killer (NK) cell infiltration into the brain significantly contributes to the exacerbation of cerebral ischemic injury, this study investigated the impact of EA at Shuigou (GV26) and Baihui (GV20) on post-ischemic brain infiltration and activation of NK cells. Methods: Neurological deficit score, rotarod test, adhesive removal test, and TTC staining were used to evaluate the beneficial effects of EA in middle cerebral artery occlusion (MCAO) mice. The inhibitory effect of EA on STAT3 activation was assessed using Western blot. Flow cytometry was used to explore the impact of EA on post-ischemic brain infiltration of NK cells, as well as the activating receptor NKG2D expression and interferon-γ (IFN-γ) production by these infiltrated NK cells. Results: EA significantly alleviated neurological functional deficits and reduced brain infarction in MCAO mice. Abundant NK cells infiltrated into the ischemic hemisphere, but this infiltration was significantly suppressed by EA. Furthermore, EA attenuated NKG2D levels and reduced production of IFN-γ by NK cells in the ischemic brain. Notably, EA's inhibitory effect on post-ischemic NK cell brain infiltration and activation was comparable to that of STAT3 inhibition. The combination of EA and STAT3 inhibition did not result in further enhancement of the inhibitory effect. Moreover, the protective effects of EA against MCAO injury were abolished when STAT3 was activated. Conclusion: Our findings suggest that EA at Shuigou (GV26) and Baihui (GV20) inhibits the post-ischemic brain infiltration and activation of NK cells through STAT3 inhibition, significantly contributing to its therapeutic effects against brain ischemia.

Abstract Multiple myeloma (MM) is an incurable hematological malignancy with poor survival. Accumulating evidence reveals that lactylation modification plays a vital role in tumorigenesis. However, research on lactylation-related genes (LRGs) in predicting the prognosis of MM remains limited. Differentially expressed LRGs (DELRGs) between MM and normal samples were investigated from the Gene Expression Omnibus database. Univariate Cox regression and LASSO Cox regression analysis were applied to construct gene signature associated with overall survival. The signature was validated in two external datasets. A nomogram was further constructed and evaluated. Additionally, Enrichment analysis, immune analysis, and drug chemosensitivity analysis between the two groups were investigated. qPCR and immunofluorescence staining were performed to validate the expression and localization of PFN1. CCK-8 and flow cytometry were performed to validate biological function. A total of 9 LRGs (TRIM28, PPIA, SOD1, RRP1B, IARS2, RB1, PFN1, PRCC, and FABP5) were selected to establish the prognostic signature. Kaplan–Meier survival curves showed that high-risk group patients had a remarkably worse prognosis in the training and validation cohorts. A nomogram was constructed based on LRGs signature and clinical characteristics, and showed excellent predictive power by calibration curve and C-index. Moreover, biological pathways, immunologic status, as well as sensitivity to chemotherapy drugs were different between high- and low-risk groups. Additionally, the hub gene PFN1 is highly expressed in MM, knocking down PFN1 induces cell cycle arrest, suppresses cell proliferation and promotes cell apoptosis. In conclusion, our study revealed that LRGs signature is a promising biomarker for MM that can effectively early distinguish high-risk patients and predict prognosis.

Abstract Acute Myeloid Leukemia (AML) is a malignant blood cancer with a high mortality rate. Neutrophil extracellular traps (NETs) influence various tumor outcomes. However, NET-related genes (NRGs) in AML had not yet received much attention. This study focuses on the role of NRGs in AML and their interaction with the immunological microenvironment. The gene expression and clinical data of patients with AML were downloaded from the TCGA-LAML and GEO cohorts. We identified 148 NRGs through the published article. Univariate Cox regression was used to analyze the association of NRGs with overall survival (OS). The least absolute shrinkage and selection operator were utilized to assess the predictive efficacy of NRGs. Kaplan–Meier plots visualized survival estimates. ROC curves assessed the prognostic value of NRG-based features. A nomogram, integrating clinical information and prognostic scores of patients, was constructed using multivariate logistic regression and Cox proportional hazards regression models. Twenty-seven NRGs were found to significantly impact patient OS. Six NRGs—CFTR, ENO1, PARVB, DDIT4, MPO, LDLR—were notable for their strong predictive ability regarding patient survival. The ROC values for 1-, 3-, and 5-year survival rates were 0.794, 0.781, and 0.911, respectively. In the training set (TCGA-LAML), patients in the high NRG risk group showed a poorer prognosis (p

For steady-state temperature field simulation during cooling stage of plastic injection molding, when the traditional finite volume method (FVM) is used to solve the equivalent heat flux on the cavity surface, the approximate node temperature gradient is often based on the average value in the control volume, but it may not be equal to the local node temperature gradient on the boundary surface. In addition, the calculation of heat flux on each surface in the control volume is independent, which may lead to the non-conservation of heat energy on the control volume. To this end, the improved FVM is modified by taking the heat conservation of steady-state temperature field as a prerequisite and calculating the heat flow of cavity surface based on the node control volume. A numerical simulation software of C++ language based on FVM is developed, which compares the steady-state temperature field of plastic injection molding simulated by the traditional FVM algorithm and the improved FVM algorithm with the results of commercial softwares. The results show that the traditional FVM algorithm can simulate the rough temperature distribution of injection mold, but there is a large calculation error. By improving the equivalent steady-state heat flux calculation method for cavity surface, the calculation error is greatly reduced, which proves that the improved FVM algorithm can predict more accurately the steady-state temperature field in cooling stage.

Abstract Background Safflower (Carthamus tinctorius L.) is an oilseed crop with substantial medicinal and economic value. However, the methods for constructing safflower core germplasm resources are limited, and the molecular mechanisms of lipid biosynthesis in safflower seeds are not well understood. Results In this study, 11 oil-related quantitative traits and 50 pairs of InDel markers were used to assess the diversity of a collection of 605 safflower germplasms. The original safflower germplasm exhibited rich phenotypic diversity, with high variation for most of the phenotypic traits under investigation. Similarly, high genetic diversity was evaluated in the original germplasm, in which the mean Shannon’s information index (I), observed heterozygosity (H 0 ), and expected heterozygosity (He) were 0.553, 0.182, and 0.374, respectively. Four subgroups with strong genetic structures were identified and a core germplasm of 214 cultivars was constructed, which is well represented in the original germplasm. Meanwhile, differential expression analysis of the transcriptomes of high and low linoleic acid safflower varieties at two stages of seed development identified a total of 47 genes associated with lipid biosynthesis. High expression of the genes KAS II and SAD enhanced the synthesis and accumulation of oleic acid, while FAD genes like FAD2 (Chr8G0104100), FAD3, FAD7 and FAD8 promoted the consumption of oleic acid conversion. The coordinated regulation of these multiple genes ensures the high accumulation of oleic acid in safflower seed oil. Conclusions Based on these findings, a core germplasm of 214 cultivars was constructed and 47 candidate genes related to unsaturated fatty acid biosynthesis and lipid accumulation were identified. These results not only provide guidance for further studies to elucidate the molecular basis of oil lipid accumulation in safflower seeds, but also contribute to safflower cultivar improvements.

Abstract Human MutT Homolog 1 (MTH1) is a nucleotide pool sanitization enzyme that hydrolyzes oxidized nucleotides to prevent their mis-incorporation into DNA under oxidative stress. Expression and functional roles of MTH1 in platelets are not known. Here, we show MTH1 expression in platelets and its deficiency impairs hemostasis and arterial/venous thrombosis in vivo. MTH1 deficiency reduced platelet aggregation, phosphatidylserine exposure and calcium mobilization induced by thrombin but not by collagen-related peptide (CRP) along with decreased mitochondrial ATP production. Thrombin but not CRP induced Ca2+-dependent mitochondria reactive oxygen species generation. Mechanistically, MTH1 deficiency caused mitochondrial DNA oxidative damage and reduced the expression of cytochrome c oxidase 1. Furthermore, MTH1 exerts a similar role in human platelet function. Our study suggests that MTH1 exerts a protective function against oxidative stress in platelets and indicates that MTH1 could be a potential therapeutic target for the prevention of thrombotic diseases.

Water electrolysis is a promising technology to produce hydrogen but it was severely restricted by the slow oxygen evolution reaction (OER). Herein, we firstly reported an advanced electrocatalyst of MOF-derived hollow Zn–Co–Ni sulfides (ZnS@Co9S8@Ni3S2-1/2, abbreviated as ZCNS-1/2) nanosword arrays (NSAs) with remarkable hydrogen evolution reaction (HER), OER and corresponding water electrolysis performance. To reach a current density of 10 mA cm−2, the cell voltage of assembled ZCNS-1/2//ZCNS-1/2 for urea electrolysis (1.314 V) is 208 mV lower than that for water electrolysis (1.522 V) and stably catalyzed for over 15 h, substantially outperforming the most reported water and urea electrolysis electrocatalysts. Density functional theory calculations and experimental result clearly reveal that the properties of large electrochemical active surface area (ECSA) caused by hollow NSAs and fast charge transfer resulted from the Co9S8@Ni3S2 heterostructure endow the ZCNS-1/2 electrode with an enhanced electrocatalytic performance.

The influence of geological development factors such as reservoir heterogeneity needs to be comprehensively considered in the determination of oil well production control strategy. In the past, many optimization algorithms are introduced and coupled with numerical simulation for well control problems. However, these methods require a large number of simulations, and the experience of these simulations is not preserved by the algorithm. For each new reservoir, the optimization algorithm needs to start over again. To address the above problems, two reinforcement learning methods are introduced in this research. A personalized Deep Q-Network (DQN) algorithm and a personalized Soft Actor-Critic (SAC)algorithm are designed for optimal control determination of oil wells. The inputs of the algorithms are matrix of reservoir properties, including reservoir saturation, permeability, etc., which can be treated as images. The output is the oil well production strategy. A series of samples are cut from two different reservoirs to form a dataset. Each sample is a square area that takes an oil well at the center, with different permeability and saturation distribution, and different oil-water well patterns. Moreover, all samples are expanded by using image enhancement technology to further increase the number of samples and improve the coverage of the samples to the reservoir conditions. During the training process, two training strategies are investigated for each personalized algorithm. The second strategy uses 4 times more samples than the first strategy. At last, a new set of samples is designed to verify the model’s accuracy and generalization ability. Results show that both the trained DQN and SAC models can learn and store historical experience, and push appropriate control strategies based on reservoir characteristics of new oil wells. The agreement between the optimal control strategy obtained by both algorithms and the global optimal strategy obtained by the exhaustive method is more than 95%. The personalized SAC algorithm shows better performance compared to the personalized DQN algorithm. Compared to the traditional Particle Swarm Optimization (PSO), the personalized models were faster and better at capturing complex patterns and adapting to different geological conditions, making them effective for real-time decision-making and optimizing oil well production strategies. Since a large amount of historical experience has been learned and stored in the algorithm, the proposed method requires only 1 simulation for a new oil well control optimization problem, which showing the superiority in computational efficiency.

Abstract Background Immune thrombocytopenia (ITP) is an autoimmune disease characterized as a low platelet count resulting from immune-mediated platelet destruction. Dimethyl fumarate (DMF) is widely applied for the treatment of several autoimmune diseases with immunosuppressive effect. However, whether it ameliorates ITP is unclear. This study aims to evaluate whether DMF has a preventive effect on ITP in mice. Methods DMF (30, 60 or 90 mg/kg body weight) was intraperitoneally injected into mice followed by injection of rat anti-mouse integrin GPIIb/CD41antibody to induce ITP. Peripheral blood was isolated to measure platelet count and spleen mononuclear cells were extracted to measure Th1 and Treg cells along with detecting the levels of IFN-γ, and TGFβ-1 in plasma and CD68 expression in spleen by immuohistochemical staining. Additionally, macrophage cell line RAW264.7 was cultured and treated with DMF followed by analysis of cell apoptosis and cycle, and the expression of FcγRI, FcγRIIb and FcγRIV mRNA. Results DMF significantly inhibited antiplatelet antibody-induced platelet destruction, decreased Th1 cells and the expression of T-bet and IFN-γ, upregulated Treg cells and the expression of Foxp3 and TGF-β1 as well as reduced CD68 expression in the spleen of ITP mouse. DMF-treated RAW264.7 cells showed S-phase arrest, increased apoptosis and downregulated expression of FcγRI and FcγRIV. Meanwhile, in vitro treatment of DMF also decreased the expression of cyclin D1 and E2, reduced Bcl-2 level and increased Bax expression and caspase-3 activation. Conclusions In conclusion, DMF prevents antibody-mediated platelet destruction in ITP mice possibly through promoting apoptosis, indicating that it might be used as a new approach for the treatment of ITP.

Most traditional injection-production optimization methods that treat the entire oil reservoir as a whole require re-optimization when facing new reservoirs, which is not only time-consuming but also does not make full use of historical experience information. This study decomposes the reservoir into independent basic production units to increase sample size and diversity and utilizes image enhancement techniques to augment the number of samples. Two frameworks based on convolutional neural networks (CNNs) are employed to recommend optimal control strategies for inputted well groups. Framework 1 uses bottom hole pressure (BHP) as a control variable and trains a CNN with optimal BHP obtained by reinforcement learning algorithms as labels. Framework 2 saves BHP and corresponding oil well revenue (NPV) during reinforcement learning optimization and trains a CNN with well groups and BHP as features and NPV as labels. The CNN in this framework is capable of directly outputting the NPV according to control strategies. The particle swarm algorithm (PSO) is used to generate control strategies and call CNN to predict development effects until PSO converges to the optimal production strategy. The experimental results demonstrate that the CNN-based frameworks outperform the traditional PSO-based methods in terms of accuracy and computational efficiency. Framework 1 achieves an output accuracy of 87% for predicting the optimal BHP for new well groups, while Framework 2 achieves an accuracy of 78%. Both frameworks exhibit fast running times, with each iteration taking less than 1 s. This study provides a more effective and accurate method for optimizing oil well production in oil reservoirs by decomposing oil reservoirs into independent units and using CNN to construct an algorithm framework, which is of great significance for the real-time optimization and control of oil wells in oil fields.

Abstract The identification of genome-wide selection signatures can provide insights on the mechanisms of natural and/or artificial selection and uncover genes related to biological functions and/or phenotypes. Tibetan sheep are an important livestock in Tibet, providing meat and wool for Tibetans who are renown for breeding livestock that adapt well to high altitudes. Using whole-genome sequences with an effective sequencing depth of 5×, we investigated the genomic diversity and structure and, identified selection signatures of White Tibetan, Oula and Poll Dorset sheep. We obtained 30,163,679 Single Nucleotide Polymorphisms (SNPs) and 5,388,372 indels benchmarked against the ovine Oar_v4.0 genome assembly. Next, using F ST, ZHp and XP-EHH approaches, we identified selection signatures spanning a set of candidate genes, including HIF1A, CAPN3, PRKAA1, RXFP2, TRHR and HOXA10 that are associated with pathways and GO categories putatively related to hypoxia responses, meat traits and disease resistance. Candidate genes and GO terms associated with coat color were also identified. Finally, quantification of blood physiological parameters, revealed higher levels of mean corpuscular hemoglobin measurement and mean corpuscular hemoglobin concentration in Tibetan sheep compared with Poll Dorset, suggesting a greater oxygen-carrying capacity in the Tibetan sheep and thus better adaptation to high-altitude hypoxia. In conclusion, this study provides a greater understanding of genome diversity and variations associated with adaptive and production traits in sheep.

Zhongwei goat is a unique Chinese native goat breed for excellent lamb fur. The pattern of flower spikes of the lamb fur was significantly reduced due to the reduction of the bending of the hair strands with growth. In order to explore the molecular mechanism underlying hair bending with growth, we performed the comprehensive analysis of transcriptome and proteome of skins from 45-days, 108-days and 365-days goat based on TMT-based quantitative proteomics and RNA-seq methods. In the three comparison groups, 356, 592 and 282 differentially expressed proteins (DEPs) were screened, respectively. KEGG pathway analysis indicated that DEPs were significantly enriched in a set of signaling pathways related to wool growth and bending, such as ECM-receptor interaction, PI3K-Akt signaling pathway, PPAR signaling pathway, protein digestion and absorption, and metabolic pathways. In addition, 20 DEPs abundance of goat skin at three development stages were examined by PRM method, which validated the reliability of proteomic data. Among them, KRT and collagen alpha family may play an important role in the development of goat hair follicle and wool bending. COL6A1, COL6A2, CRNN, TNC and LOC102178129 were identified as candidate genes based on combined analysis of transcriptome and proteome data and PRM quantification. Our results identify the differential expressed proteins as well as pathways related to the wool bending of Zhongwei goats and provide a theoretical basis for further revealing the molecular mechanism underlying wool bending of goats.

Cancer, as one of the most life-threatening diseases, has attracted the attention of researchers to develop drugs with minimal side effects. The bioactive macromolecules, such as the polysaccharides, are considered the potential candidates against cancer due to their anti-tumor activities and non-toxic characteristics. The present review provides an overview on polysaccharides' extraction, isolation, purification, mechanisms for their anti-tumor activities, structure-activity relationships, absorption and metabolism of polysaccharides, and the applications of polysaccharides in anti-tumor therapy. Numerous research showed extraction methods of polysaccharides had a significant influence on their activities. Additionally, the anti-tumor activities of the polysaccharides are closely related to their structure, while molecular modification and high bioavailability may enhance the anti-tumor activity. Moreover, most of the polysaccharides exerted an anti-tumor activity mainly through the cell cycle arrest, anti-angiogenesis, apoptosis, and immunomodulation mechanisms. Also, recommendations were made to utilize the polysaccharides against cancer.

After large-scale sand fracturing of horizontal wells in shale gas reservoir, fracturing fractures will deform in the production process. However, fracture deformation will lead to the decrease in fracture conductivity and then cause the decrease of gas well productivity. Therefore, in order to evaluate the fracturing fracture deformation mechanism of shale reservoirs, the shale proppant-supported fracture deformation evaluation experiments were carried out under different proppant types, particle sizes, sanding concentrations, and closure pressure conditions, respectively, and the variation curves of fracture width was calculated by a stereomicroscope under different experimental conditions. Then based on the experimental results, the fracture sensitivity factors and fracture deformation mechanism were analyzed, and the deformation mechanisms of fracturing fractures affected by proppant embedding and crushing were studied emphatically. The analysis results of fracture sensitivity factors indicate that the larger the particle size and hardness of proppant, the lower the sand concentration, proppant embedded on the shale rock surface. Moreover, the deeper the proppant is embedded, the faster the fracture conductivity decreases. In addition, the greater the closure pressure, the more serious is the proppant embedment, and the faster the fracture width decreases. The analysis results of fracture deformation mechanism show that, on the on hand, under variable closure pressure, the proppant with larger hardness and larger particle size is used for fracturing, and the proppant embedded in the fracture surface is the main cause of fracture deformation. However, if only the sand concentration of the proppant in the fracture is changed, the fracture deformation is jointly dominated by the embedding and crushing of the proppant. On the other hand, under constant closure pressure, the main mechanism of fracture deformation is that the proppant is embedded into the fracture surface when the closure pressure is low, but if the closure pressure is high, the main mechanism of fracture deformation is the crushing and compaction of proppant.

Abstract Background Elevated red blood cell distribution width (RDW) and decreased platelet count (PLT) can be clinically relevant to the prognosis in cancer patients. However, their prognostic values in patients with diffuse large B-cell lymphoma (DLBCL) need to be further explored. Methods Healthy donors (n = 130) and patients with DLBCL (n = 349) were included and evaluated retrospectively in this study. The prognostic influence of clinical and pathological factors including RDW and PLT on overall survival (OS) and progression-free survival (PFS) were studied by Kaplan-Meier curves. To evaluate the independent prognostic relevance of RDW and PLT, univariate and multivariate Cox proportional hazards regression models were applied. The adjusted IPI model was established based on the results of multivariate analysis, and verified by Harrell’s C statistical analysis. Results Kaplan-Meier curves indicated that an elevated RDW value and thrombocytopenia are poor factors for OS (P

Matrine is a naturally occurring alkaloid and possesses a wide range of pharmacological properties, such as anti-cancer, anti-oxidant, anti-inflammatory effects. However, whether it affects platelet function and thrombosis remains unclear. This study aims to evaluate the effect of matrine on platelet function and thrombus formation. Human platelets were treated with matrine (0–1 mg/ml) for 1 h at 37°C followed by measuring platelet aggregation, granule secretion, receptor expression by flow cytometry, spreading and clot retraction. In addition, matrine (10 mg/kg) was injected intraperitoneally into mice to measure tail bleeding time, arterial and venous thrombus formation. Matrine dose-dependently inhibited platelet aggregation and ATP release in response to either collagen-related peptide (Collagen-related peptide, 0.1 μg/ml) or thrombin (0.04 U/mL) stimulation without altering the expression of P-selectin, glycoprotein Ibα, GPVI, or αIIbβ3. In addition, matrine-treated platelets presented significantly decreased spreading on fibrinogen or collagen and clot retraction along with reduced phosphorylation of c-Src. Moreover, matrine administration significantly impaired the in vivo hemostatic function of platelets, arterial and venous thrombus formation. Furthermore, in platelets stimulated with CRP or thrombin, matrine significantly reduced Reactive oxygen species generation, inhibited the phosphorylation level of ERK1/2 (Thr202/Tyr204), p38 (Thr180/Tyr182) and AKT (Thr308/Ser473) as well as increased VASP phosphorylation (Ser239) and intracellular cGMP level. In conclusion, matrine inhibits platelet function, arterial and venous thrombosis, possibly involving inhibition of ROS generation, suggesting that matrine might be used as an antiplatelet agent for treating thrombotic or cardiovascular diseases.

In this work, the methane desorption isothermal curves at different water contents on deep sampled from Western Chongqing of China were measured at pressures up to 65 MPa and at 130°C by the volumetric method. In the first instance, the desorption increases with the decrease of pressure, the adsorbed gas desorbs slightly with decreasing pressures from 65 to 30 MPa. When the pressure drops to 30–20 MPa, the desorption rate increases rapidly with the decrease of pressure and the desorption curve begins to separate from the adsorption curve, resulting in desorption hysteresis. At last, when the pressure is lower than 20 MPa, the desorption increases almost linearly with the further decrease of pressure, but eventually there will be some adsorbed gas which cannot be desorbed to form residual adsorbed gas. After that, the isotherm desorption data of CH4 was fitted using the improved desorption model. The fitting results showed that the improved desorption model can be used to describe the desorption process of deep shale gas containing water and has a strong applicability. In addition, the critical desorption pressure increases with increasing water content. When the water content is lower than 1%, the effect of the water content on the desorption of deep shale gas increases rapidly with increasing water content, as well as when the water content is greater than 1%, the impact changes slowly.

NADPH oxidase-derived reactive oxygen species (ROS) regulates platelet function and thrombosis. It remains controversial regarding NOX2’s role in platelet function. As a regulatory subunit for NOX2, whether p47phox regulates platelet function remains unclear. Our study intends to evaluate p47phox’s role in platelet function. Platelets were isolated from wild-type or p47phox-/- mice followed by analysis of platelet aggregation, granule secretion, surface receptors expression, spreading, clot retraction and ROS generation. Additionally, in vivo hemostasis, arterial and venous thrombosis was assessed. Moreover, human platelets were treated with PR-39 to inhibit p47phox activity followed by analysis of platelet function. p47phox deficiency significantly prolonged tail-bleeding time, delayed arterial and venous thrombus formation in vivo as well as reduced platelet aggregation, ATP release and αIIbβ3 activation. In addition, p47phox-/- platelets presented impaired spreading on fibrinogen or collagen and defective clot retraction concomitant with decreased phosphorylation of Syk and PLCγ2. Moreover, CRP or thrombin-stimulated p47phox-/- platelets displayed reduced intracellular ROS generation which was further decreased after inhibition of NOX1. Meanwhile, p47phox deficiency increased VASP phosphorylation and decreased phosphorylation of ERK1/2, p38, ERK5 and JNK without affecting AKT and c-PLA2 phosphorylation. Furthermore, p47phox translocates to membrane to interact with both NOX1 and NOX2 after stimulation with CRP or thrombin. Finally, inhibition of p47phox activity by PR-39 reduced ROS generation, platelet aggregation and clot retraction in human platelets. In conclusion, p47phox regulates platelet function, arterial and venous thrombus formation and ROS generation, indicating that p47phox might be a novel therapeutic target for treating thrombotic or cardiovascular diseases.

Three immunoglobulin (Ig) isotypes have been identified in teleosts, IgM, IgD, and IgT or IgZ. IgT, a new teleost Ig isotype, plays a vital role in mucosal immunity. However, information on molecular and functional characteristics of fish IgT is still limited. In this study, an IgT heavy chain (LcIgT) gene was cloned and characterized in large yellow croaker (Larimichthys crocea). Complete cDNA of LcIgT was 1930 bp in length, encoding a protein of 554 amino acids. The deduced LcIgT contains a VH region and only three CH regions (CH1, CH2, CH4), but no transmembrane region was predicted. Phylogenetic analysis showed that IgT heavy chain sequences from all fish species are grouped together. Homology comparison showed that LcIgT shares the highest amino acid identity of 58.73% with IgT heavy chain in Scophthalmus maximus. The VH domain of LcIgT has the highest identity of 72.50% with that of Scophthalmus maximus IgT. Relatively, each constant domain of LcIgT exhibits the highest amino acid identity with that of IgT in Oreochromis niloticus (67.61% identity for CH1, 61.11% identity for CH2, and 63.74% identity for CH4). LcIgT was constitutively expressed in various tissues tested, with the highest levels in mucosa-associated tissues such as gills and skin. After Cryptocaryon irritans infection, the mRNA levels of LcIgT were significantly up-regulated in the spleen (3.27-fold) at 4 d, in the head kidney (3.98-fold) and skin (2.11-fold) at 7 d, and in gills (4.45-fold) at 14 d. The protein levels in these detected tissues were all significantly up-regulated; the peak of its up-regulation was 6.33-fold at 28d in gills, 3.44-fold at 7d in skin, and 3.72-fold at 14d in spleen. These results showed that IgT response could be simultaneously induced in both systemic and mucosal tissues after parasitic infection and that IgT may be involved in systemic immunity and mucosal immunity against parasitic infection.

The Zhongwei goat is kept primarily for its beautiful white, curly pelt that appears when the kid is approximately 1 month old; however, this representative phenotype often changes to a less curly phenotype during postnatal development in a process that may be mediated by multiple molecular signals. DNA methylation plays important roles in mammalian cellular processes and is essential for the initiation of hair follicle (HF) development. Here, we sought to investigate the effects of genome-wide DNA methylation by combining expression profiles of the underlying curly fleece dynamics. Genome-wide DNA methylation maps and transcriptomes of skin tissues collected from 45- to 108-day-old goats were used for whole-genome bisulfite sequencing (WGBS) and RNA sequencing, respectively. Between the two developmental stages, 1,250 of 3,379 differentially methylated regions (DMRs) were annotated in differentially methylated genes (DMGs), and these regions were mainly related to intercellular communication and the cytoskeleton. Integrated analysis of the methylome and transcriptome data led to the identification of 14 overlapping genes that encode crucial factors for wool fiber development through epigenetic mechanisms. Furthermore, a functional study using human hair inner root sheath cells (HHIRSCs) revealed that, one of the overlapping genes, platelet-derived growth factor C (PDGFC) had a significant effect on the messenger RNA expression of several key HF-related genes that promote cell migration and proliferation. Our study presents an unprecedented analysis that was used to explore the enigma of fleece morphological changes by combining methylome maps and transcriptional expression, and these data revealed stage-specific epigenetic changes that potentially affect fiber development. Furthermore, our functional study highlights a possible role for the overlapping gene PDGFC in HF cell growth, which may be a predictable biomarker for fur goat selection.