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When conducting condition recognition research on AC contactor vibration signals through time-frequency analysis, the feature data exhibit a high degree of redundancy, which leads to repetitive information and hinders the accuracy of recognition. To address the redundancy issue in the features of AC contactor vibration signals, this study introduces a feature selection method based on Regularized Random Forest with Recursive Selection (RFRS). Initially, a test platform for AC contactor vibration signals was established, and time-frequency domain features of the AC contactor vibration signals were extracted. Subsequently, the traditional Random Forest (RF) was refined by optimizing its stopping criteria using the Recursive Feature Elimination approach and by incorporating a regularization coefficient during the splitting process to direct the split towards significant features. This modification not only enhances the Random Forest's capacity to leverage existing information but also introduces a bias, enabling it to favor important features. Finally, through case analysis, the proposed method effectively reduced the dimensionality of the feature set and achieved an average of 87.37% for Recall, 87.41% for F1-Score, 88.38% for Precision, and 85.74% for Accuracy. The overall performance of this method surpasses that of the three mainstream feature selection methods: Spearman's rank correlation coefficient method, the embedded method, and the filter method. This study thus provides a rather effective feature selection approach for the state recognition study of AC contactors., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Diabetes is a chronic disease, which is characterized by abnormally high blood sugar levels. It may affect various organs and tissues, and even lead to life-threatening complications. Accurate prediction of diabetes can significantly reduce its incidence. However, the current prediction methods struggle to accurately capture the essential characteristics of nonlinear data, and the black-box nature of these methods hampers its clinical application. To address these challenges, we propose KCCAM_DNN, a diabetes prediction method that integrates Kendall's correlation coefficient and an attention mechanism within a deep neural network. In the KCCAM_DNN, Kendall's correlation coefficient is initially employed for feature selection, which effectively filters out key features influencing diabetes prediction. For missing values in the data, polynomial regression is utilized for imputation, ensuring data completeness. Subsequently, we construct a deep neural network (KCCAM_DNN) based on the self-attention mechanism, which assigns greater weight to crucial features affecting diabetes and enhances the model's predictive performance. Finally, we employ the SHAP model to analyze the impact of each feature on diabetes prediction, augmenting the model's interpretability. Experimental results show that KCCAM_DNN exhibits superior performance on both PIMA Indian and LMCH diabetes datasets, achieving test accuracies of 99.090% and 99.333%, respectively, approximately 2% higher than the best existing method. These results suggest that KCCAM_DNN is proficient in diabetes prediction, providing a foundation for informed decision-making in the diagnosis and prevention of diabetes., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Qi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Copper (Cu) homeostasis are important processes in the cause of metabolic diseases, but the association between Cu and obesity remains unclear., Methods: Participants were drawn from the 2011-2016 National Health and Nutrition Examination Survey (NHANES). Weighted logistic regression assessed the associations of serum Cu concentrations (tertiles) with obesity and central obesity in individuals without comorbidities. Obesity was defined as a BMI ≥30.0 kg/m2, and central obesity was defined as a waist circumference ≥80 cm for women and ≥95 cm for men., Results: This cross-sectional study included 1,665 adults without comorbidities, representing 24,744,034 people (mean age 35.1 years, 48.5% female). High serum Cu levels (tertile 3: ≥19.19 μmol/L) were associated with higher odds of obesity (adjusted odds ratio [OR]: 4.48, 95% CI[confidence interval]: 2.44-8.32) and central obesity (OR: 2.36, 95% CI: 1.19-4.66) compared to low serum Cu levels (tertile 1: ≤15.64 μmol/L). The dose-response curve showed a nonlinear association between Cu levels and obesity (P-nonlinear = 0.02) and a linear association with central obesity (P-nonlinear = 0.21)., Conclusion: This study suggests that higher serum Cu levels are associated with increased odds of obesity in healthy American adults., Competing Interests: NHANES is a comprehensive cross-sectional study conducted in the United States. The survey employs a sophisticated, stratified, and multistage probability sampling design. The research protocol received approval from the Research Ethics Review Board of the National Center for Health Statistics, and NHANES ensures that written informed consent is obtained from all participants. It’s important to note that our analysis exclusively utilized publicly available data, ensuring the absence of personally identifiable information. All procedures strictly adhered to relevant guidelines and regulations. The interview questionnaires and examination response rates are accessible to the public. This does not alter our adherence to PLOS ONE policies on sharing data and materials., (Copyright: © 2024 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

This study investigates the dynamic relationship between cross-border e-commerce and logistics co-development strategies within the context of small and medium-sized enterprises (SMEs) in China. The primary objective is to formulate an empirical model capable of estimating the utility and flexibility of cross-border e-commerce logistics, specifically focusing on its role in achieving competitive advantage for SMEs. The research employs a comprehensive approach, considering various factors that influence the formulation and implementation of cross-border e-commerce logistics strategies. Factors such as scale, quality, potential, and infrastructure are scrutinized to provide a nuanced understanding of the dynamic interplay. Real-world data is analyzed using advanced statistical techniques to derive meaningful insights. The data of CBEC and logistics planning in Guangdong from 2013 to 2018 was used as a case example to demonstrate its and flexibility and usefulness of empirically proposed model estimation and develop a holistic indicator system for the evaluation of the synergistic development of CBEC and logistics. This research focus on the modelling of the CBEC for the synergistic effect analysis and evaluate the usefulness and flexibility of CBEC to achieve competitive advantage. The study aims to uncover insights into the most effective approaches for Chinese SMEs in navigating the landscape of cross-border e-commerce. By examining the results derived from the empirical model, the research sheds light on the impact of cross-border e-commerce logistics on competitive advantage. The findings contribute to a deeper understanding of how SMEs can strategically position themselves in the cross-border e-commerce arena. Through the analysis of real-world data and the application of advanced statistical methods, this research offers valuable insights for Chinese SMEs. The insights generated from the study not only illuminate the intricacies of cross-border e-commerce logistics but also provide practical recommendations to reduce associated costs. Ultimately, the study aims to equip SMEs with the knowledge necessary to thrive in the evolving landscape of cross-border e-commerce., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Qi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

As a UNESCO World Cultural Heritage, the aesthetic value of bronze artifacts from the Shang and Chow Dynasties has had a profound influence on Chinese traditional culture and art. To facilitate the digital preservation and protection of these Shang and Chow bronze artifacts (SCB), it becomes imperative to categorize their decorative patterns. Therefore, a SCB pattern classification method of differential evolution called Shang and Chow Bronze Convolutional Neural Network (SCB-CNN) is proposed. Firstly, the original bronze decorative patterns of Shang and Chow dynasties are collected, and the samples are expanded through image augmentation technology to form a training dataset. Secondly, based on the classical convolutional neural network structure, the recognition and classification of bronze patterns are implemented by adjusting the network parameters. Then, the initial parameters of the convolutional neural network are optimized by differential evolution algorithm, and the optimized SCB-CNN is simulated. Finally, comparative experiments were conducted between the optimized SCB-CNN, the unoptimized model, VGG-Net, and GoogleNet. The experimental results indicate that the optimized SCB-CNN significantly reduces training time while maintaining fast prediction speed, convergence speed, and high accuracy. This study provides new insights for the inheritance and innovation research of SCB patterns., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Qi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

How to evaluate the resilience level and change trend of supply chain is an important research direction in current supply chain management practice. This paper proposes a new method of supply chain resilience assessment based on hesitant fuzzy linguistic term set (HFLTS) and matter element extension theory. Firstly, based on the research status quo at home and abroad, a low-carbon enterprise supply chain resilience assessment index system is established, which includes six first-level indicators and corresponding 21 second-level indicators of product supply resilience, resource resilience, partner resilience, information response resilience, financial resilience and knowledge resilience. Secondly, HFLTS was used to collect expert opinions and Ordered Weighted Arithmetic (OWA) to calculate the expert composite language, by which the fuzzy evaluation matrix of supply chain resilience assessment indicators was obtained. Once again, the resilience indicator weights are determined based on a game-theoretic portfolio assignment method combining the best-worst method (BWM) and the CRITIC method. Finally, the nearness degree function is combined with the extension comprehensive evaluation method to improve the matter element extension model, and the supply chain resilience assessment model of low-carbon enterprises based on the game theory combination assignment-improved matter element extension is established. Taking X low-carbon enterprise as an example, the evaluation results show that the supply chain resilience level of this enterprise is II, and the eigenvalue of the grade variable is 2.69, and the supply chain resilience is shifting to III, and the supply chain resilience is shifting to III, which indicates that the supply chain resilience of this enterprise is being enhanced. Therefore, the improved matter element extension not only ensures the accuracy of the evaluation results, but also has higher prediction accuracy., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Luo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

One of the basic assumptions of spatial theory is formulated in Waldo Tobler's first law of geography: "everything is related to everything else, but near things are more related than distant things." However, as internet space is a complex virtual space independent of the real world, whether this law is applicable to things in the internet space remains to be explored in depth. Therefore, this study takes the event of Asian elephant northern migration as an example, attempts to investigate the issue of the applicability of Tobler's first law of geography to internet attention by integrating geographic methods such as spatial visualization, spatial correlation analysis, and Geo-detector. The results show that Tobler's first law of geography does not fully apply to internet attention, which does not decay with increasing distance. Geographical distance, within certain boundaries, is influenced by "identity" and "relevance", and still plays a large role in internet attention. However, once the boundaries are exceeded, the impact of geographic distance on internet attention is weakened by the intervention of influencing factors such as the degree of information technology, population, and the strength of news media publicity. Overall, the strength of news media publicity has the greatest impact on internet attention. And when it interacts with geographic proximity, it has the most significant effect on internet attention., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Zheng et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: A case-control study was conducted to evaluate the relationship between endothelial nitric oxide synthase (NOS3) gene polymorphism and essential hypertension in the Han, Miao, and Buyi populations in Guizhou China., Methods: DNA was collected from the blood samples of 353 essential hypertension patients and 342 healthy controls from Guizhou province of China. Eight polymorphisms of the NOS3 gene were genotyped using the Sequenom MassARRAY platform. For genetic analysis, SPSS 26.0, Haploview, SNPStats, SHEsis, and MDR were utilized., Results: All SNPs (rs11771443, rs1808593, rs753482, rs3918186, rs3918188, rs3918227, rs7830, and rs891512) satisfied the Hardy-Weinberg equilibrium test (P > 0.05). The allele and genotype frequencies of rs7830 and rs1808593 in case-control groups demonstrated significant differences (P < 0.05). Compared to the TT genotype of rs1808593, the TG or GG genotype reduced the risk of hypertension in the Miao population (OR = 0.410, 95% CI: 0.218-0.770, P = 0.006). Compared to the GG or GT genotype of rs7830, the TT genotype increased the risk of hypertension in the overall populations (OR = 1.716, 95%CI: 1.139-2.586, P = 0.010). The CATT (rs3918227-rs391818186-rs1808593-rs7830) haplotype was a risk factor for hypertension in the Miao and Han populations (OR = 1.471, 95%CI: 1.010-2.143, P = 0.044 and OR = 1.692, 95%CI: 1.124-2.545, P = 0.011). The CAGG haplotype in the Miao population was a protective factor against hypertension (OR = 0.555, 95%CI: 0.330-0.934, P = 0.025). The rs3918188, rs1808593, and rs7830 in the Miao population showed an interaction effect on hypertension (P < 0.001). The rs11771443, rs3918188, and rs7830 in the Buyi and Han populations showed an interaction effect on hypertension (P = 0.013 and P < 0.001)., Conclusion: The single nucleotide polymorphisms rs1808593 and rs7830 of NOS3 gene are associated with essential hypertension in Guizhou ethnic populations., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The unsaturated permeability coefficients are often used to solve geotechnical problems associated with unsaturated soils. But it is very difficult to measure. However, the unsaturated permeability coefficients can be predicted by the Soil-water Characteristic Curves (SWCCs). The Van Genuchten Model (VG model) is very rife as it's smooth and good fitting, thus, it has the most research data. Therefore, the research data on VG model parameters (α, n, θs and θr) of Malan loess in Chinese Loess Plateau are collected in the past two decades to obtain the spatial distribution characteristics of parameters. The trend surface analysis method is employed to clarify the regional scale distribution and the variation regular pattern on ArcGIS. Then the linear regression method is utilized to fit the relationship between suction and water content in three different regions of Chinese Loess Plateau, which is divided according to the properties and particle gradation. By using this relationship and the trend surface analysis contour map, the unsaturated permeability coefficient of the sample can be predicted after measuring the saturated permeability coefficient. The example verification shows that the difference between the prediction results and the experimental results is very small when the sample has the lower saturation, and the deviation is slightly larger if it has the higher saturation, but they are all within the acceptable range. This method not only saves the test cost, but also considers the physical properties of the loess in the three different regions of the Loess Plateau. With the improvement of data and the gradual improvement of sampling density, the prediction accuracy will gradually improve. It can provide convenience for solving the engineering problems of loess and water and other engineering applications., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Fang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objective: When facing major emergency public accidents, men and women may react differently. Our research aimed to assess the influence of gender difference on social support, information preference, biological rhythm, psychological distress, and the possible interaction among these factors during the COVID-19 pandemic., Methods: In this cross-sectional study, 3,237 respondents aged 12 years and older finished the online survey. Levels of social support, information preference, biological rhythm, and psychological distress were assessed using validated scales. A path analysis was conducted to explore possible associations among these variables., Results: The path analysis indicated that women with high levels of social support had a lower possibility of biological rhythm disorders and lower levels of somatization symptoms of psychological distress during the COVID-19 pandemic. The influence of social support on somatization symptoms was exerted via biological rhythm. Women tended to believe both negative and positive information, while men preferred more extreme information., Conclusion: Our results highlighted gender difference in study variables during the COVID-19 pandemic and the importance of social support in alleviating psychological distress and biological rhythm disorders. Moreover, we confirmed that information preference differed significantly by somatization symptoms of psychological distress, suggesting extra efforts to provide more individualized epidemic information. Longitudinal research is required to further explore casual inferences., Competing Interests: The authors have declared that no competing interests exist.

Zika virus (ZIKV) is a positive-sense RNA flavivirus and can cause serious neurological disorders including microcephaly in infected fetuses. As a mosquito-borne arbovirus, it enters the bloodstream and replicates in various organs. During pregnancy, it can be transmitted from the blood of the viremic mother to the fetus by crossing the placental barrier. Monocytes and macrophages are considered the earliest blood cell types to be infected by ZIKV. As a first line defense, these cells are crucial components in innate immunity and host responses and may impact viral pathogenesis in humans. Previous studies have shown that ZIKV infection can activate inflammasomes and induce proinflammatory cytokines in monocytes. In this report, we showed that ZIKV could infect and induce cell death in human and murine macrophages. In addition to the presence of cleaved caspase-3, indicating that apoptosis was involved, we identified the cleaved caspase-1 and gasdermin D (GSDMD) as well as increased secretion of IL-1β and IL-18. This suggests that the inflammasome was activated and that may lead to pyroptosis in infected macrophages. The pyroptosis was NLRP3-dependent and could be suppressed in the macrophages treated with shRNA to target and knockdown caspase-1. It was also be inhibited by an inhibitor for caspase-1, indicating that the pyroptosis was triggered via a canonical approach. Our findings in this study demonstrate a concomitant occurrence of apoptosis and pyroptosis in ZIKV-infected macrophages, with two mechanisms involved in the cell death, which may have potentially significant impacts on viral pathogenesis in humans., Competing Interests: The authors have declared that no competing interests exist.

After long-term oxidation and energy storage, broken coal body borehole walls and drainage shaft walls may cause spontaneous combustion during gas extraction. The high-temperature thermal shock caused by the spontaneous combustion of coal incurs thermal damage on adjacent coal, which, in turn, causes changes in the mechanical properties of the coal. However, only a few studies have been conducted in this context, which has limited our understanding of the thermal damage characteristics of coal bodies in such situations. This study aimed to experimentally investigate the correlation between the crack evolution law and the mechanical properties of coal bodies at different temperatures (50-300°C) using heat-force loading considering Ping Mei No. 10 coal mine as the research object. The results suggest that the coal body experiences a large amount of visible damage, and becomes increasingly complex. At 50-300°C, some indexes (such as longitudinal wave velocity, Poisson's ratio, compressive strength, elastic modulus, impact energy index, and pre-peak strain) are positively correlated with temperature. In addition, the dynamic failure time and temperature show a negative correlation, and the overall change slope is small. The relationship between each index and temperature at 200-300°C is opposite to that at 50-200°C, and the overall change slope is larger. Moreover, when the oxidation temperature exceeds 200°C, the destruction of the coal body changes from elastic brittleness to ductility-plasticity. High-temperature oxidation incurs irreversible thermal damage of coal. Hence, it is necessary to focus on the changes in mechanical properties of coal after a spontaneous combustion process is extinguished., Competing Interests: The authors have declared that no competing interests exist.

Prefabricated construction has attracted worldwide concern and promotion due to its environmental friendliness, high quality, and high efficiency. In China, the application of prefabricated construction still lags due to its high cost. To improve prefabricated construction development, the Chinese government and provinces have launched subsidy policies for different objects that offer subsidies to the assembler, the manufacturer, or consumers. Subsidy policies for different subsidy objects have different impacts on the manufacturer wholesale price and assembler retail price and assembly rate and make their decisions more complicated. Therefore, this study uses game theory and builds three models to analyze the effects of government subsidies on manufacturer pricing, assembler pricing, assembly rate decisions, and profit. We find that government subsidy policies can bring more profit to prefabricated construction enterprises, reduce their costs, and benefit the promotion of prefabricated construction. Through comparison and numerical analysis, we also find that when the government subsidizes enterprises more, it is better to subsidize the assembler, because it is good for all three parties. First, consumers can obtain a lower retail price. Second, enterprises can obtain more profits. Finally, for the government, this approach can increase the demand for prefabricated construction and increase the assembly rate, which is conducive to the promotion of prefabricated construction. When the government subsidizes customers more, it is better for the assembler and the manufacturer to subsidize customers, because they can obtain more profits. It is better for the government and customers to subsidize the assembler or the manufacture, because consumers can get the lower retail price. Although the assembly rate and enterprises' profits are not optimal, they have also been improved. In addition, when the government directly subsidizes enterprises, the enterprises will actively cooperate with the subsidy policy and are more willing to adopt prefabricated construction. This approach will benefit the promotion of prefabricated construction., Competing Interests: The authors have declared that no competing interests exist.

Aconitine (AC) is the primary bioactive and secondary metabolite alkaloidin of Aconitum species which is accounted for more than 60% of the total diester-diterpenoid alkaloids in Aconite. To evaluate the analgesic effects of AC, 4 different pain models including hot plate assay, acetic acid writhing assay, formalin and CFA induced pain models were adopted in this study. In hot plate experiment, AC treatment at concentration of 0.3 mg/kg and 0.9 mg/kg improved the pain thresholds of mice similar to the positive drug aspirin at the concentration of 200 mg/kg (17.12% and 20.27% VS 19.21%). In acetic acid writhing experiment, AC significantly reduced the number of mice writhing events caused by acetic acid, and the inhibition rates were 68% and 76%. These results demonstrated that AC treatment revealed significant analgesic effects in both acute thermal stimulus pain model and chemically-induced visceral pain model. The biphasic nociceptive responses induced by formalin were significantly inhibited after AC treatment for 1h or 2h. The inhibition rates were 33.23% and 20.25% of AC treatment for 1h at 0.3 mg/kg and 0.9 mg/kg in phase I. In phase II, the inhibition rates of AC and aspirin were 36.08%, 32.48% and 48.82% respectively, which means AC showed similar analgesic effect to non-steroidal anti-inflammatory compounds. In the chronic CFA-induced nociception model, AC treatment also improved mice pain threshold to 131.33% at 0.3 mg/kg, which was similar to aspirin group (152.03%). Above all, our results verified that AC had obviously analgesic effects in different mice pain models., Competing Interests: The authors have declared that no competing interests exist.

Lycium ruthenicum is an excellent eco-economic shrub. Numerous researches have been conducted for the function of its fruits but scarcely focused on the somaclonal variation and DNA methylation. An efficient micropropagation protocol from leaves and stems of L. ruthenicum was developed in this study, in which not only the leaf explants but also the stem explants of L. ruthenicum were dedifferentiated and produced adventitious buds/multiple shoots on one type of medium. Notably, the efficient indirect organogenesis of stem explants was independent of exogenous auxin, which is contrary to the common conclusion that induction and proliferation of calli is dependent on exogenous auxin. We proposed that sucrose supply might be the crucial regulator of stem callus induction and proliferation of L. ruthenicum. Furthermore, results of methylation-sensitive amplified polymorphism (MSAP) showed that DNA methylation somaclonal variation (MSV) of CNG decreased but that of CG increased after acclimatization. Three types of micropropagated plants (from leaf calli, stem calli and axillary buds) were epigenetically diverged more from each other after acclimatization and the ex vitro micropropagated plants should be selected to determine the fidelity. In summary, plants micropropagated from axillary buds and leaves of L. ruthenicum was more fidelity and might be suitable for preservation and propagation of elite germplasm. Also, leaf explants should be used in transformation. Meanwhile, plants from stem calli showed the highest MSV and might be used in somaclonal variation breeding. Moreover, one MSV hotspot was found based on biological replicates. The study not only provided foundations for molecular breeding, somaclonal variation breeding, preservation and propagation of elite germplasm, but also offered clues for further revealing novel mechanisms of both stem-explant dedifferentiation and MSV of L. ruthenicum., Competing Interests: The authors have declared that no competing interests exist.

The endoplasmic reticulum (ER) membrane protein complex (EMC) is a conserved protein complex involved in inserting the transmembrane domain of membrane proteins into membranes in the ER. EMC3 is an essential component of EMC and is important for rhodopsin synthesis in photoreceptor cells. However, the in vivo function of Emc3 in bipolar cells (BCs) has not been determined. To explore the role of Emc3 in BCs, we generated a BC-specific Emc3 knockout mouse model (named Emc3 cKO) using the Purkinje cell protein 2 (Pcp2) Cre line. Although normal electroretinography (ERG) b-waves were observed in Emc3 cKO mice at 6 months of age, Emc3 cKO mice exhibited reduced b-wave amplitudes at 12 months of age, as determined by scotopic and photopic ERG, and progressive death of BCs, whereas the ERG a-wave amplitudes were preserved. PKCa staining of retinal cryosections from Emc3 cKO mice revealed death of rod BCs. Loss of Emc3 led to the presence of the synaptic protein mGLuR6 in the outer nuclear layer (ONL). Immunostaining analysis of presynaptic protein postsynaptic density protein 95 (PSD95) revealed rod terminals retracted to the ONL in Emc3 cKO mice at 12 months of age. In addition, deletion of Emc3 resulted in elevated glial fibrillary acidic protein, indicating reactive gliosis in the retina. Our data demonstrate that loss of Emc3 in BCs leads to decreased ERG response, increased astrogliosis and disruption of the retinal inner nuclear layer in mice of 12 months of age. Taken together, our studies indicate that Emc3 is not required for the development of BCs but is important for long-term survival of BCs., Competing Interests: The authors have declared that no competing interests exist.

[This corrects the article DOI: 10.1371/journal.pone.0065381.].

Among the genus Actinidia, Actinidia arguta possesses the strongest cold resistance and produces fresh fruit with an intense flavor. To investigate genomic variation that may contribute to variation in phenotypic traits, we performed whole-genome re-sequencing of four A. arguta genotypes originating from different regions in China and identified the polymorphisms using InDel markers. In total, 4,710,650, 4,787,750, 4,646,026, and 4,590,616 SNPs and 1,481,002, 1,534,198, 1,471,304, and 1,425,393 InDels were detected in the 'Ruby-3', 'Yongfeng male', 'Kuilv male', and 'Hongbei male' genomes, respectively, compared with the reference genome sequence of cv 'Hongyang'. A subset of 120 InDels were selected for re-sequencing validation. Additionally, genes related to non-synonymous SNPs and InDels in coding domain sequences were screened for functional analysis. The analysis of GO and KEGG showed that genes involved in cellular responses to water deprivation, sucrose transport, decreased oxygen levels and plant hormone signal transduction were significantly enriched in A. arguta. The results of this study provide insight into the genomic variation of kiwifruit and can inform future research on molecular breeding to improve cold resistance in kiwifruit., Competing Interests: The authors have declared that no competing interests exist.

The One-to-one Pickup and Delivery Problem with Shortest-path Transport along Real-life Paths (OPDPSTRP) is presented in this paper. It is a variation of the One-to-one Pickup and Delivery Problem (OPDP), which is common in daily life, such as the Passenger Train Operation Plans (PTOP) and partial Taxi-sharing Problem. Unlike the classical OPDP, in the OPDPSTRP, (1) each demand must be transported along the shortest path according to passengers/shippers requirements, and (2) each vehicle should travel along a real-life path. First, six route structure rules are proposed for the OPDPSTRP, and a kind of Mixed-Integer Programming (MIP) models is formulated for it. Second, A Variable Neighborhood Descent (VND), a Variable Neighborhood Research (VNS), a Multi-Start VND (MS&#95;VND) and a Multi-Start VNS (MS&#95;VNS) with five neighborhood operators has been developed to solve the problem. Finally, The Gurobi solver, the VND, the VNS, the MS&#95;VND and the MS&#95;VNS have been compared with each other by 84 random instances partitioned in small size connected graphs, medium size connected graphs and large size connected graphs. From the test results we found that solutions generated by these approaches are often comparable with those found by the Gurobi solver, and the solutions found by these approaches are better than the solutions found by the Gurobi solver when solving instances with larger numbers of demands. In almost all instances, the MS&#95;VND significantly outperforms the VND and the VNS in terms of solution quality, and outperforms the MS&#95;VNS both in terms of solution quality and CPU time. In the instances with large numbers of demands, the MS&#95;VND is still able to generate good feasible solutions in a reasonable CPU time, which is of vital practical significance for real-life instances., Competing Interests: The authors have declared that no competing interests exist.

Tick-borne encephalitis (TBE) is endemic to Europe and some Asian countries and is prevalent in northeast China. We analyzed the epidemiology of TBE in China from 2007 to 2018. A total of 3,364 TBE cases were reported in mainland China from 2007 to 2018, for an annual incidence of 0.09 to 0.44/100,000. Among the TBE cases, 89.92% were reported in forest areas (41.94% in DaXingAnLing, 8.70% in XiaoXingAnLing, and 39.21% in ChangBaiShan) in northeast China. The TBE cases were primarily male with a proportion of 67.15% (2,259/3,364 cases) and in 40-49-year age group with a proportion of 31.89% (1,073/3,364 cases). The epidemiology of TBE differed slightly among the three forest regions. Domestic workers and forestry workers accounted for the most of the TBE cases in DaXingAnLing, and domestic workers and farmers in XiaoXingAnLing and ChangBaiShan, respectively. The TBE cases mainly occurred from April to August with a peak in June. The TBE laboratory confirmed rate in DaXingAnLing (84.14%, 1,189/1,413 cases) was highest, compared with XiaoXingAnLing and ChangBaiShan (13.99% and 11.37%, respectively). Moreover, the hospital with the highest laboratory confirmed rate (88.01%, 1,336/1,518 cases) was Inner Mongolia Forestry General Hospital of DaXingAnling region. Systematic enhanced TBE surveillance and a vaccination program are needed to improve the laboratory confirmed rate and reduce the incidence of TBE in northeast China., Competing Interests: The authors have declared that no competing interests exist.

As important regulators, miRNAs could play pivotal roles in regulation of fruit coloring. Actinidia arguta is a newly emerged fruit tree with extensively application prospects. However, miRNAs involved in A. arguta fruit coloring are unknown. In this study, A. arguta fruit were investigated at three developmental stages by small RNAs high-throughput sequencing. A total of 482 conserved miRNAs corresponding to 526 pre-miRNAs and 581 novel miRNAs corresponding to 619 pre-miRNAs were grouped into 46 miRNA families. Target gene prediction and analysis revealed that miR858, a strongly candidate miRNA, was involved in anthocyanin biosynthesis in which contributes to fruit coloring. The anthocyanin level was determined in three A. arguta cultivars by UPLC-MS/MS (ultra-performance liquid chromatography coupled with tandem mass spectrometry). In addition, qPCR (quantitative real-time PCR), cluster analysis were conducted as well as correlation analysis. All results were combined to propose a model in which describes an association of miRNA and anthocyanin biosynthesis in A. arguta. The data presented herein is the first report on miRNA profile analysis in A. arguta, which can provide valuable information for further research into the regulation of the miRNAs in anthocyanin biosynthesis and fruit coloring., Competing Interests: The authors have declared that no competing interests exist.

Emerging evidences indicate that hepatitis B virus (HBV) infection is associated with non-Hodgkin lymphoma (NHL), but the mechanisms of HBV-induction lymphomagenesis remain unclear. In this report, retrospective analysis of the prevalence of hepatitis B surface antigen (HBsAg) among NHL cases demonstrated significantly higher HBsAg carrier rate among B-cell NHL cases than controls (other cancers except primary liver cancer) (adjusted odds ratio, 1.56; 95% confidence interval, 1.13-2.16). Furthermore, cells with an immortalization potential existed in the peripheral blood of 4 patients with chronic HBV infection. Characterization of these cells showed their immunophenotypes similar to that of the majority of HBsAg-positive B-cell NHL patients. Immunoglobulin (Ig) gene rearrangements confirmed the clonal Ig gene rearrangements. Cytogenetic analysis revealed abnormal karyotypes of these cells with an immortalization potential. Compared with cells with an immortalization potential that we previously found in B-cell NHL patients by the same way, these cells showed many similar features. In conclusion, cells with an immortalization potential existed in the part of patients with chronic HBV infection before lymphoma development and showed some malignant features. They may be the cellular basis of HBV-associated lymphomagenesis., Competing Interests: The authors have declared that no competing interests exist.

Kiwifruit bacterial canker is a devastating disease threatening kiwifruit production. To clarify the defense mechanism in response to Pseudomonas syringae pv. actinidiae (Psa), we observed phenotypic changes in resistant Huate (HT) and susceptible Hongyang (HY) kiwifruit varieties at 0, 12, 24, 48, 96, and 144 hour after inoculation (hai) with Psa. Brown lesions appeared in the inoculation areas 12 hai in HY shoots, and the lesion length gradually increased from 24 to 144 h. In contrast, no lesions were found in HT shoots at any time points. Furthermore, RNA-seq analysis showed significantly more differentially expressed genes between HT and HY at 12 hai than at any other time point. According to weighted gene co-expression network analysis, five modules were notably differentially expressed between HT and HY; pathway mapping using the Kyoto Encyclopedia of Gene and Genomes database was performed for the five modules. In MEgreenyellow and MEyellow modules, pathways related to"plant-pathogen interaction", "Endocytosis", "Glycine, serine and threonine metabolism", and "Carbon fixation in photosynthetic organisms" were enriched, whereas in the MEblack module, pathways related to "protein processing in endoplasmic reticulum", "plant-pathogen interaction", and "Glycolysis / Gluconeogenesis" were enriched. In particular, the Pti1 and RPS2 encoding effector receptors, and the NPR1, TGA, and PR1 genes involved in the salicylic acid signaling pathway were significantly up-regulated in HT compared with HY. This indicates that the effector-triggered immunity response was stronger and that the salicylic acid signaling pathway played a pivotal role in the Psa defense response of HT. In addition, we identified other important genes, involved in phenylpropanoid biosynthesis and Ca2+ internal flow, which were highly expressed in HT. Taken together, these results provide important information to elucidate the defense mechanisms of kiwifruit during Psa infection., Competing Interests: The authors have declared that no competing interests exist.

Chicken anaemia virus (CAV), a member of the genus Gyrovirus, is the etiological agent of chicken infectious anaemia. CAV infects bone marrow-derived cells, resulting in severe anaemia and immunosuppression in young chickens and a compromised immune response in older birds. We investigated the molecular epidemiology of CAV in sick chickens in China from 2014 to 2015 and showed that the CAV-positive rate was 13.30%, in which mixed infection (55.56%) was the main type of infection. We isolated and identified 15 new CAV strains using different methods including indirect immunofluorescence assay and Western Blotting. We used overlapping polymerase chain reaction to map the whole genome of the strains. Phylogenetic analyses of the obtained sequences and related sequences available in GenBank generated four distinct groups (A-D). We built phylogenetic trees using predicted viral protein (VP) sequences. Unlike CAV VP2s and VP3s that were well conserved, the diversity of VP1s indicated that the new strains were virulent. Our epidemiological study provided new insights into the prevalence of CAV in clinical settings in recent years in China., Competing Interests: The authors have declared that no competing interests exist.

Neovascularization is the pathological driver of blinding eye diseases such as retinopathy of prematurity, proliferative diabetic retinopathy, and wet age-related macular degeneration. The loss of vision resulting from these diseases significantly impacts the productivity and quality of life of patients, and represents a substantial burden on the health care system. Current standard of care includes biologics that target vascular endothelial growth factor (VEGF), a key mediator of neovascularization. While anti-VGEF therapies have been successful, up to 30% of patients are non-responsive. Therefore, there is a need for new therapeutic targets, and small molecule inhibitors of angiogenesis to complement existing treatments. Apelin and its receptor have recently been shown to play a key role in both developmental and pathological angiogenesis in the eye. Through a cell-based high-throughput screen, we identified 4-aminoquinoline antimalarial drugs as potent selective antagonists of APJ. The prototypical 4-aminoquinoline, amodiaquine was found to be a selective, non-competitive APJ antagonist that inhibited apelin signaling in a concentration-dependent manner. Additionally, amodiaquine suppressed both apelin-and VGEF-induced endothelial tube formation. Intravitreal amodaiquine significantly reduced choroidal neovascularization (CNV) lesion volume in the laser-induced CNV mouse model, and showed no signs of ocular toxicity at the highest doses tested. This work firmly establishes APJ as a novel, chemically tractable therapeutic target for the treatment of ocular neovascularization, and that amodiaquine is a potential candidate for repurposing and further toxicological, and pharmacokinetic evaluation in the clinic., Competing Interests: A patent covering the use of the compounds for pathological angiogenesis was submitted to the USPTO (“4-aminoquinoline compounds for the treatment of angiogenesis”; filed March 6, 2018; application number 62/639,291). This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Actinidia arguta is the most basal species in a phylogenetically and economically important genus in the family Actinidiaceae. To better understand the molecular basis of the Actinidia arguta chloroplast (cp), we sequenced the complete cp genome from A. arguta using Illumina and PacBio RS II sequencing technologies. The cp genome from A. arguta was 157,611 bp in length and composed of a pair of 24,232 bp inverted repeats (IRs) separated by a 20,463 bp small single copy region (SSC) and an 88,684 bp large single copy region (LSC). Overall, the cp genome contained 113 unique genes. The cp genomes from A. arguta and three other Actinidia species from GenBank were subjected to a comparative analysis. Indel mutation events and high frequencies of base substitution were identified, and the accD and ycf2 genes showed a high degree of variation within Actinidia. Forty-seven simple sequence repeats (SSRs) and 155 repetitive structures were identified, further demonstrating the rapid evolution in Actinidia. The cp genome analysis and the identification of variable loci provide vital information for understanding the evolution and function of the chloroplast and for characterizing Actinidia population genetics., Competing Interests: The authors have declared that no competing interests exist.

The selection of seismic attributes is a key process in reservoir prediction because the prediction accuracy relies on the reliability and credibility of the seismic attributes. However, effective selection method for useful seismic attributes is still a challenge. This paper presents a novel selection method of seismic attributes for reservoir prediction based on the gray relational degree (GRD) and support vector machine (SVM). The proposed method has a two-hierarchical structure. In the first hierarchy, the primary selection of seismic attributes is achieved by calculating the GRD between seismic attributes and reservoir parameters, and the GRD between the seismic attributes. The principle of the primary selection is that these seismic attributes with higher GRD to the reservoir parameters will have smaller GRD between themselves as compared to those with lower GRD to the reservoir parameters. Then the SVM is employed in the second hierarchy to perform an interactive error verification using training samples for the purpose of determining the final seismic attributes. A real-world case study was conducted to evaluate the proposed GRD-SVM method. Reliable seismic attributes were selected to predict the coalbed methane (CBM) content in southern Qinshui basin, China. In the analysis, the instantaneous amplitude, instantaneous bandwidth, instantaneous frequency, and minimum negative curvature were selected, and the predicted CBM content was fundamentally consistent with the measured CBM content. This real-world case study demonstrates that the proposed method is able to effectively select seismic attributes, and improve the prediction accuracy. Thus, the proposed GRD-SVM method can be used for the selection of seismic attributes in practice.

Density functional theory was used to study the effects of different types of oxygen-containing functional groups on the adsorption of oxygen molecules and single active oxygen atoms on carbonaceous materials. During gasification or combustion reactions of carbonaceous materials, oxygen-containing functional groups such as hydroxyl(-OH), carbonyl(-CO), quinone(-O), and carboxyl(-COOH) are often present on the edge of graphite and can affect graphite's chemical properties. When oxygen-containing functional groups appear on a graphite surface, the oxygen molecules are strongly adsorbed onto the surface to form a four-member ring structure. At the same time, the O-O bond is greatly weakened and easily broken. The adsorption energy value indicates that the adsorption of oxygen molecules changes from physisorption to chemisorption for oxygen-containing functional groups on the edge of a graphite surface. In addition, our results indicate that the adsorption energy depends on the type of oxygen-containing functional group. When a single active oxygen atom is adsorbed on the bridge site of graphite, it gives rise to a stable epoxy structure. Epoxy can cause deformation of the graphite lattice due to the transition of graphite from sp2 to sp3 after the addition of an oxygen atom. For quinone group on the edge of graphite, oxygen atoms react with carbon atoms to form the precursor of CO2. Similarly, the single active oxygen atoms of carbonyl groups can interact with edge carbon atoms to form the precursor of CO2. The results show that oxygen-containing functional groups on graphite surfaces enhance the activity of graphite, which promotes adsorption on the graphite surface.

p62 is a scaffolding adaptor implicated in the clearance of protein aggregates by autophagy. Reactive oxygen species (ROS) can either stimulate or inhibit NFκB-mediated gene expression influencing cellular fate. We studied the effect of hydrogen peroxide (H2O2)-mediated oxidative stress and NFκB signaling on p62 expression in the retinal pigment epithelium (RPE) and investigated its role in regulation of autophagy and RPE survival against oxidative damage. Cultured human RPE cell line ARPE-19 and primary human adult and fetal RPE cells were exposed to H2O2-induced oxidative stress. The human apolipoprotein E4 targeted-replacement (APOE4) mouse model of AMD was used to study expression of p62 and other autophagy proteins in the retina. p62, NFκB p65 (total, phosphorylated, nuclear and cytoplasmic) and ATG10 expression was assessed by mRNA and protein analyses. Cellular ROS and mitochondrial superoxide were measured by CM-H2DCFDA and MitoSOX staining respectively. Mitochondrial viability was determined using MTT activity. qPCR-array system was used to investigate autophagic genes affected by p62. Nuclear and cytoplasmic levels of NFκB p65 were evaluated after cellular fractionation by Western blotting. We report that p62 is up-regulated in RPE cells under H2O2-induced oxidative stress and promotes autophagic activity. Depletion of endogenous p62 reduces autophagy by downregulation of ATG10 rendering RPE more susceptible to oxidative damage. NFκB p65 phosphorylation at Ser-536 was found to be critical for p62 upregulation in response to oxidative stress. Proteasome inhibition by H2O2 causes p62-NFκB signaling as antioxidant pre-treatment reversed p62 expression and p65 phosphorylation when RPE was challenged by H2O2 but not when by Lactacystin. p62 protein but not RNA levels are elevated in APOE4-HFC AMD mouse model, suggesting reduction of autophagic flux in disease conditions. Our findings suggest that p62 is necessary for RPE cytoprotection under oxidative stress and functions, in part, by modulating ATG10 expression. NFκB p65 activity may be a critical upstream initiator of p62 expression in RPE cells under oxidative stress.

Sex differences in spatial memory have long been observed in humans, non-human primates and rodents, but the underlying cellular and molecular mechanisms responsible for these differences remain obscure. In the present study we found that adolescent male rats outperformed female rats in 7 d and 28 d retention probes, but not in learning trials and immediate probes, in the Morris water maze task. Male rats also had larger long-term potentiation (LTP) at hippocampal temproammonic-CA1 (TA-CA1) synapses, which have been implicated to play a key role in place field and memory consolidation, when protocols designed to elicit late-stage LTP (LLTP) were used. Interestingly, the ratio of evoked AMPA/NMDA currents was found to be smaller at TA-CA1 synapses in male rats compared to female rats. Protein biotinylation experiments showed that male rats expressed more surface GluN1 receptors in hippocampal CA1 stratum lacunosum-moleculare (SLM) than female rats, although GluA1 expression was also slightly higher in male rats. Taken together, our results suggest that differences in the expression of AMPA and NMDA receptors may affect LTP expression at TA-CA1 synapses in adolescent male and female rats, and thus possibly contribute to the observed sex difference in spatial memory., Competing Interests: The authors have declared that no competing interests exist.

Hepcidin, one kind of antimicrobial peptides, is one of the promising alternatives to antibiotics with broad spectrum of antimicrobial activity. Hepcidins cloned from different kinds of fishes have been produced using exogenous expression systems, and their in vitro antimicrobial effects have been verified. However their in vivo effects on gut microbiota and gut health of hosts remain unclear. Here we performed a safety study of hepcidin so that it can be used to reduce microbial contaminations in the food and feed. In this study, Pichia pastoris-expressed Pseudosciaena crocea hepcidin (PC-hepc) was first assessed by simulated digestion tests and then administered to male and female Sprague-Dawley (SD) rats in different concentrations. Subchronic toxicity testing, high throughput 16S rRNA sequencing of gut microbiota, and examinations on gut metabolism and permeability were conducted. The results showed PC-hepc could be digested in simulated intestinal fluid but not in simulated gastric fluid. PC-hepc had no adverse effects on general health, except causing increase of blood glucose (still in the normal value range of this index) in all trial groups of female rats and intestinal inflammation in HD group of female rats. Community composition of gut microbiota of female MD and HD groups shifted compared with control group, of which the decrease of genus Akkermansia might be related to the increase of blood glucose and intestinal inflammation. Significant increase of fecal nitroreductase activity was also observed in female MD and HD groups. Our results suggest the uses of exogenous PC-hepc in normal dosage are safe, however excess dosage of it may cause intestinal disorder of animals., Competing Interests: The authors have declared that no competing interests exist.

Cucumber is an economically important vegetable crop worldwide. Powdery mildew (PM) is one of the most severe diseases that can affect cucumber crops. There have been several research efforts to isolate PM resistance genes for breeding PM-resistant cucumber. In the present study, we used a chromosome segment substitution line, SSL508-28, which carried PM resistance genes from the donor parent, JIN5-508, through twelve generations of backcrossing with a PM-susceptible inbred line, D8. We performed whole-genome resequencing of SSL508-28 and D8 to identify single nucleotide polymorphisms (SNPs), and insertions and deletions (indels). When compared against the reference genome of the inbred cucumber line 9930, a total of 468,616 SNPs and 67,259 indels were identified in SSL508-28, and 537,352 SNPs and 91,698 indels were identified in D8. Of these, 3,014 non-synonymous SNPs and 226 frameshift indels in SSL508-28, and 3,104 non-synonymous SNPs and 251 frameshift indels in D8, were identified. Bioinformatics analysis of these variations revealed a total of 15,682 SNPs and 6,262 indels between SSL508-28 and D8, among which 120 non-synonymous SNPs and 30 frameshift indels in 94 genes were detected between SSL508-28 and D8. Finally, out of these 94 genes, five resistance genes with nucleotide-binding sites and leucine-rich repeat domains were selected for qRT-PCR analysis. This revealed an upregulation of two transcripts, Csa2M435460.1 and Csa5M579560.1, in SSL508-28. Furthermore, the results of qRT-PCR analysis of these two genes in ten PM resistant and ten PM susceptible cucumber lines showed that when exposed to PM, Csa2M435460.1 and Csa5M579560.1 exhibited a higher expression level of resistant lines than susceptible lines. This indicates that Csa2M435460.1 and Csa5M579560.1 are candidate genes for PM resistance in cucumber. In addition, the non-synonymous SNPs in Csa2M435460.1 and Csa5M579560.1, identified in SSL508-28 and D8, might be the key to high PM-resistance in SSL508-28., Competing Interests: The authors have declared that no competing interests exist.

Lung cancer remains the leading cancer killer around the world. It's crucial to identify newer mechanism-based targets to effectively manage lung cancer. Annexin A5 (ANXA5) is a protein kinase C inhibitory protein and calcium dependent phospholipid-binding protein, which may act as an endogenous regulator of various pathophysiological processes. However, its molecular mechanism in lung cancer remains poorly understood. This study was designed to determine the mechanism of ANXA5 in lung cancer with a hope to obtain useful information to provide a new therapeutic target. We used a stable isotope dimethyl labeling based quantitative proteomic method to identify differentially expressed proteins in NSCLC cell lines after ANXA5 transfection. Out of 314 proteins, we identified 26 and 44 proteins that were down- and up-regulated upon ANXA5 modulation, respectively. The IPA analysis revealed that glycolysis and gluconeogenesis were the predominant pathways modulated by ANXA5. Multiple central nodes, namely HSPA5, FN1, PDIA6, ENO1, ALDOA, JUP and KRT6A appeared to occupy regulatory nodes in the protein-protein networks upon ANXA5 modulation. Taken together, ANXA5 appears to have pleotropic effects, as it modulates multiple key signaling pathways, supporting the potential usefulness of ANXA5 as a potential target in lung cancer. This study might provide a new insight into the mechanism of ANXA5 in lung cancer., Competing Interests: The authors have declared that no competing interests exist.

Platelet to lymphocyte ratio (PLR) is a parameter reflecting inflammatory responses in patients with cancer. Several studies have investigated the prognostic value of PLR in patients with colorectal cancer (CRC); however, the results are controversial. Thus, we carried out a meta-analysis to evaluate the association between PLR and CRC prognostication. Relevant articles were retrieved through PubMed, Embase, and Web of Science, and pooled hazard ratio (HR) and 95% confidence interval (CI) were computed by using STATA V.12.0. Both the random-effects model and fixed-effects model were utilized. A total of 13 studies (14 cohorts) with 8,601 patients were included in the meta-analysis. Pooled HRs and 95% CIs demonstrated that increased PLR predicted poor overall survival (OS) (HR = 1.81, 95%CI:1.42-2.31, p<0.001; I2 = 65%, Ph = 0.002), disease-free survival (DFS) (HR = 1.84, 95%CI:1.22-2.76, p = 0.003; I2 = 78.3%, Ph<0.001) and recurrence-free survival (RFS) (HR = 1.84, 95%CI:1.41-2.41, p<0.001; I2 = 0, Ph = 0.686), although this was not the case for cancer-specific survival (CSS) (HR = 1.75, 95%CI:0.59-5.17, p = 0.309; I2 = 66.2%, Ph = 0.085) or time to recurrence (TTR) (HR = 1.21 95%CI:0.62-2.36, p = 0.573;I2 = 58.4%, Ph = 0.121). Subgroup analysis showed that PLR enhanced the prognostic value for OS in Caucasian patients, in small sample studies and for metastatic disease; however, this was not the case with rectal cancer. Furthermore, elevated PLR predicted reduced DFS in Caucasians and not in Asians. In conclusion, our meta-analysis showed that high PLR was a significant biomarker for poor OS, DFS, and RFS in patients with CRC; however, it had no association with CSS or TTR., Competing Interests: The authors have declared that no competing interests exist.

The molecular mechanism regulated by microRNAs (miRNAs) that underlies postnatal hypertrophy of skeletal muscle is complex and remains unclear. Here, the miRNAomes of longissimus dorsi muscle collected at five postnatal stages (60, 120, 150, 180, and 210 days after birth) from Large White (commercial breed) and Min pigs (indigenous breed of China) were analyzed by Illumina sequencing. We identified 734 miRNAs comprising 308 annotated miRNAs and 426 novel miRNAs, of which 307 could be considered pig-specific. Comparative analysis between two breeds suggested that 60 and 120 days after birth were important stages for skeletal muscle hypertrophy and intramuscular fat accumulation. A total of 263 miRNAs were significantly differentially expressed between two breeds at one or more developmental stages. In addition, the differentially expressed miRNAs between every two adjacent developmental stages in each breed were determined. Notably, ssc-miR-204 was significantly more highly expressed in Min pig skeletal muscle at all postnatal stages compared with its expression in Large White pig skeletal muscle. Based on gene ontology and KEGG pathway analyses of its predicted target genes, we concluded that ssc-miR-204 may exert an impact on postnatal hypertrophy of skeletal muscle by regulating myoblast proliferation. The results of this study will help in elucidating the mechanism underlying postnatal hypertrophy of skeletal muscle modulated by miRNAs, which could provide valuable information for improvement of pork quality and human myopathy.

Cannabis is the most widely used illicit drug in the world. Delta-9-tetrahydrocannabinol (Δ9-THC) is the main psychoactive component of cannabis and its effects have been well-studied. However, cannabis contains many other cannabinoids that affect brain function. Therefore, these studies investigated the effect of cannabis smoke exposure on locomotor activity, rearing, anxiety-like behavior, and the development of dependence in rats. It was also investigated if cannabis smoke exposure leads to tolerance to the locomotor-suppressant effects of the endogenous cannabinoid anandamide. Cannabis smoke was generated by burning 5.7% Δ9-THC cannabis cigarettes in a smoking machine. The effect of cannabis smoke on the behavior of rats in a small and large open field and an elevated plus maze was evaluated. Cannabis smoke exposure induced a brief increase in locomotor activity followed by a prolonged decrease in locomotor activity and rearing in the 30-min small open field test. The cannabinoid receptor type 1 (CB1) receptor antagonist rimonabant increased locomotor activity and prevented the smoke-induced decrease in rearing. Smoke exposure also increased locomotor activity in the 5-min large open field test and the elevated plus maze test. The smoke exposed rats spent more time in the center zone of the large open field, which is indicative of a decrease in anxiety-like behavior. A high dose of anandamide decreased locomotor activity and rearing in the small open field and this was not prevented by rimonabant or pre-exposure to cannabis smoke. Serum Δ9-THC levels were 225 ng/ml after smoke exposure, which is similar to levels in humans after smoking cannabis. Exposure to cannabis smoke led to dependence as indicated by more rimonabant-precipitated somatic withdrawal signs in the cannabis smoke exposed rats than in the air-control rats. In conclusion, chronic cannabis smoke exposure in rats leads to clinically relevant Δ9-THC levels, dependence, and has a biphasic effect on locomotor activity.

Hyperosmolarity has been recognized as an important pathological factor in dry eye leading to ocular discomfort and damage. As one of the major neuropeptides of corneal innervation, substance P (SP) has been shown to possess anti-apoptotic effects in various cells. The aim of this study was to determine the capacity and mechanism of SP against hyperosmotic stress-induced apoptosis in cultured corneal epithelial cells. The cells were exposed to hyperosmotic stress by the addition of high glucose in the presence or absence of SP. The results showed that SP inhibited hyperosmotic stress-induced apoptosis of mouse corneal epithelial cells. Moreover, SP promoted the recovery of phosphorylated Akt level, mitochondrial membrane potential, Ca2+ contents, intracellular reactive oxygen species (ROS) and glutathione levels that impaired by hyperosmotic stress. However, the antiapoptotic capacity of SP was partially suppressed by Akt inhibitor or glutathione depleting agent, while the neurokinin-1 (NK-1) receptor antagonist impaired Akt activation and ROS scavenging that promoted by SP addition. In conclusion, SP protects corneal epithelial cells from hyperosmotic stress-induced apoptosis through the mechanism of Akt activation and ROS scavenging via the NK-1 receptor.

Background: The HIV-epidemic among MSM in China has worsened. In this key population, prevalence of HSV-2 and syphilis infection and co-infection with HIV is high., Methods: A longitudinal study was conducted (n = 962) in Beijing, China, with three overlapping cohorts (n = 857, 757 and 760) consisting of MSM that were free from pairs of infections of concern (i.e. HIV-HSV-2, HIV-syphilis, HSV-2-syphilis) at baseline to estimate incidence of HIV, HSV-2, syphilis, and those of co-infection., Results: The incidence of HIV, HSV-2 and syphilis in the overall cohort was 3.90 (95% CI = 2.37, 5.43), 7.87 (95% CI = 5.74, 10.00) and 6.06 (95% CI = 4.18, 7.94) cases per 100 person-years (PYs), respectively. The incidence of HIV-HSV-2, HIV-Syphilis and HSV-2-Syphilis co-infections was 0.30 (95% CI = 0.29, 0.88), 1.02 (95% CI = 0.13, 2.17) and 1.41 (95% CI: 0.04, 2.78) cases per 100 PYs, respectively, in the three sub-cohorts constructed for this study., Conclusions: The incidence of HIV, HSV-2 and syphilis was very high and those of their co-infections were relatively high. Such co-infections have negative impacts on the HIV/STI epidemics. Prevention practices need to take such co-infections into account.

Pak choi is a seed vernalization-type plant whose vernalization mechanism is currently unclear. Therefore, it is critical to discover genes related to vernalization and research its functions during vernalization in pak choi. Here, the gene expression profiles in the shoot apex were analyzed after low temperature treatment using high-throughput RNA sequencing technology. The results showed that there are 1,664 and 1,192 differentially expressed genes (DEGs) in pak choi in cold treatment ending and before flower bud differentiation, respectively, including 42 genes that exhibited similar expression trend at both stages. Detailed annotation revealed that the proteins encoded by the DEGs are located in the extracellular region, cell junction and extracellular matrix. These proteins exhibit activity such as antioxidant activity and binding protein/transcription factor activity, and they are involved in signal transduction and the immune system/biological processes. Among the DEGs, Bra014527 was up-regulated in low temperature treatment ending, Bra024097 was up-regulated before flower bud differentiation and Bra035940 was down-regulated at both stages in low temperature-treated shoot apices. Homologues of these genes in A. thaliana, AT3G59790, AT4G30200 and AT5G61150, are involved in flowering and vernalization, suggesting that they take part in the vernalization process in pak choi. Further pathway enrichment analysis revealed that most genes were enriched in the tryptophan metabolism and glucosinolate biosynthesis pathways. However, the functions of tryptophan and glucosinolate in vernalization are not yet clear and require further analysis.

Mulberry, belonging to the order Rosales, family Moraceae, and genus Morus, has received attention because of both its economic and medicinal value, as well as for its important ecological function. The genus Morus has a worldwide distribution, however, its taxonomy remains complex and disputed. Many studies have attempted to classify Morus species, resulting in varied numbers of designated Morus spp. To address this issue, we used information from internal transcribed spacer (ITS) genetic sequences to study the taxonomy of all the members of generally accepted genus Morus. We found that intraspecific 5.8S rRNA sequences were identical but that interspecific 5.8S sequences were diverse. M. alba and M. notabilis showed the shortest (215 bp) and the longest (233 bp) ITS1 sequence length, respectively. With the completion of the mulberry genome, we could identify single nucleotide polymorphisms within the ITS locus in the M. notabilis genome. From reconstruction of a phylogenetic tree based on the complete ITS data, we propose that the Morus genus should be classified into eight species, including M. alba, M. nigra, M. notabilis, M. serrata, M. celtidifolia, M. insignis, M. rubra, and M. mesozygia. Furthermore, the classification of the ITS sequences of known interspecific hybrid clones into both paternal and maternal clades indicated that ITS variation was sufficient to distinguish interspecific hybrids in the genus Morus.

Blumeria graminis f. sp. tritici, which causes wheat powdery mildew, is an obligate biotrophic pathogen that can easily genetically adapt to its host plant. Understanding the virulence structure of and genetic variations in this pathogen is essential for disease control and for breeding resistance to wheat powdery mildew. This study investigated 17 pathogenic populations in Sichuan, China and classified 109 isolates into two distinct groups based on pathogenicity analysis: high virulence (HV, 92 isolates) and low virulence (LV, 17 isolates). Populations from Yibin (Southern region), Xichang (Western region), and Meishan (Middle region) showed lower virulence frequencies than populations from other regions. Many of the previously known resistance genes did not confer resistance in this study. The resistance gene Pm21 displayed an immune response to pathogenic challenge with all populations in Sichuan, and Pm13, Pm5b, Pm2+6, and PmXBD maintained resistance. AMOVA revealed significantly higher levels of variation within populations and lower levels of variation among populations within regions. High levels of gene flow were detected among populations in the four regions. Closely related populations within each region were distinguished by cluster analyses using ISSR and SRAP alleles. Both ISSR and SRAP allele profiling analyses revealed high levels of genetic diversity among pathogenic populations in Sichuan. Although ISSR and SRAP profiling analysis showed similar resolutions, the SRAP alleles appeared to be more informative. We did not detect any significant association between these alleles and the virulence or pathogenicity of the pathogen. Our results suggest that ISSR and SRAP alleles are more efficient for the characterization of small or closely related populations versus distantly related populations.

Background: Peripheral artery disease (PAD), which is caused by atherosclerosis, results in progressive narrowing and occlusion of the peripheral arteries and inhibits blood flow to the lower extremities. Therapeutic angiogenesis is a promising strategy for treating ischemia caused by PAD. Nitric oxide (NO) has been shown to be a key mediator of angiogenesis. It has been demonstrated that β-cyclodextrincan stimulate vessel growth in rabbit corneas. In this study, we assessed the mechanism of action and therapeutic potential of a new angiogenic molecule, (2-hydroxypropyl)-β-cyclodextrin (2HP-β-CD)., Methods and Results: 2HP-β-CD significantly increased vascular endothelial growth factor A (VEGF-A) and platelet-derived growth factor BB (PDGF-BB) peptides in human umbilical vein endothelial cells (HUVECs) and also increased basic fibroblast growth factor (bFGF) peptide in human aortic smooth muscle cells (HASMCs). 2HP-β-CD stimulated both proliferation and migration of HUVECs in an endothelial nitric oxide synthase (eNOS)/NO-dependent manner, whereas NO was found to be involved in proliferation, but not migration, of HASMCs. In a unilateral hindlimb ischemia model in mice, 2HP-β-CD injections not only promoted blood flow recovery and increased microvessel densities in ischemic muscle, but also promoted coverage of the vessels with smooth muscle cells, thus stabilizing the vessels. Administration of 2HP-β-CD increased the expression of several angiogenic factors, including VEGF-A, PDGF-BB and transforming growth factor beta-1 (TGF-β1) in ischemic muscle. Injections of 2HP-β-CD also stimulated protein kinase B and extracellular regulated protein kinases (ERK), leading to an increase in phosphorylation of eNOS in ischemic muscle. Treatment with the NOS inhibitor, Nω-nitro-L-arginine methyl ester (L-NAME), showed that stimulation of blood flow induced by 2HP-β-CD was partially dependent on NO., Conclusions: Therapeutic angiogenesis by 2HP-β-CD may be beneficial to patients with PAD.

Background and Aim: The coagulation abnormalities in non-cirrhotic Budd-Chiari syndrome (NC-BCS) and non-cirrhotic portal vein thrombosis (NC-PVT) are unclear. We conducted this case-control study to investigate the coagulation profile of NC-BCS and NC-PVT in Chinese patients., Methods: We measured the levels of factors II, V, VII, VIII, IX, X, XI, XII, protein C (PC), protein S (PS) and antithrombin (AT) in blood samples from 37 NC-BCS patients, 74 NC-PVT patients, and 100 healthy controls. The levels and ratios of pro- and anti-coagulation factors were compared between patients with NC-BCS and healthy controls, between different types of NC-BCS and between NC-PVT and healthy controls., Results: In patients with NC-BCS, factor VIII (P<0.001) was significantly elevated; factor V (P<0.001), VII (P<0.001), IX (P = 0.003), X (P<0.001), XI (P<0.001), XII (P<0.001), PC (P<0.001) and AT (P<0.001) were significantly decreased; and no difference was observed for factor II (P = 0.088) and PS (P = 0.199) compared with healthy controls. Factor VIII-to-PC (P = 0.008), factor VIII-to-PS (P = 0.037) and factor VIII-to-AT (P = 0.001) were significantly increased; other ratios were significantly reduced or did not show any difference. No differences were observed between different types of NC-BCS for individual pro- and anti-coagulation factors or the ratios between them. Among patients with NC-PVT, factor VIII (P<0.001) was significantly elevated and other factors were significantly decreased. Factor II-to-PC (P<0.001), factor VIII-to-PC (P<0.001), factor IX-to-PC (P<0.001), factor VIII-to-PS (P<0.001), factor II-to-AT (P<0.001), factor VIII-to-AT (P<0.001) and factor IX-to-AT (P<0.001) were significantly increased; all other ratios for NC-PVT were significantly reduced or did not show any significant difference., Conclusions: NC-BCS and NC-PVT are associated with elevated levels of factor VIII and the decreased levels of PC and AT were probably the most significant features of coagulation imbalance. Additionally, NC-PVT was associated with decreased levels of PS.

Background: Influenza H7N9 and H1N1pdm can cause severe human infections. It is important to investigate the distinguishing clinical features between these two diseases. Several studies have compared the differences in general, however, age and gender adjusted comparisons may be more useful and informative to the health professionals., Methods: A total of 184 severe H1N1pdm patients and 37 severe H7N9 patients from Jiangsu Province were included in this analysis to perform age and gender adjusted comparison of clinical features., Results: After adjusting age and gender, no significant differences in chronic medical conditions or treatment were found between severely ill patients with H7N9 and H1N1pdm. Severely ill patients with H7N9 had significantly longer interval from onset of illness to neuraminidase inhibitor treatment and to death. They were more likely to have complications such as acute respiratory distress syndrome (ARDS), liver and renal dysfunctions, and had a significantly higher risk of death., Conclusion: Our results suggests that age and gender should be adjusted as important confounding factors when comparing the clinical features between severe H7N9 and H1N1pdm patients to avoid any misunderstanding regarding the differences between these two diseases particularly in terms of clinical severity and prognosis.

Plant-specific NAC proteins are one of the largest families of transcription factors in plants, and members of this family have been characterized with roles in the regulation of diverse biological processes, including development and stress responses. In the present study, we identified 101 putative NAC domain-encoding genes (BdNACs) through systematic sequence analysis in Brachypodium distachyon, a new model plant of family Poaceae. BdNAC proteins were phylogenetically clustered into 13 groups, and each group possesses similar motif compositions. Phylogenetic analysis using known stress-related NACs from Arabidopsis and rice as query sequences identified 18 BdNACs as putative stress-responsive genes. In silico promoter analysis showed that almost all BdNAC genes contain putative stress-related cis-elements in their promoter regions. Expression profile of BdNAC genes in response to abiotic stresses and phytohormones was analyzed by quantitative real-time RT-PCR. Several putative stress-responsive BdNAC genes, including BdNAC003 and BdNAC044 which is ortholog of known stress-responsive rice gene SNAC1 and SNAC2, respectively, were highly regulated by multiple abiotic stresses and stress-related phytohormone treatments. Taken together, our results presented here would be helpful in laying the foundation for understanding of the complex mechanisms of NAC mediated abiotic stress signaling transduction pathways in B. distachyon.

Epidermal growth factor (EGF) has been shown to facilitate the in vitro maturation of sheep oocytes, and enhance embryo's capability for further development. However, such kind of molecular mechanism has not yet been elucidated. In the present study, we investigated the effect of EGF-mediated mitogen-activated protein kinases 3 and 1 (MAPK3/1) pathway on in vitro maturation of sheep oocytes. U0126, a specific inhibitor of MEK (MAPK kinase), was added into the maturation culture medium to block the EGF-mediated MAPK3/1 pathway with different doses. Then, the nuclear maturation of sheep oocytes was examined. Additionally, the effect of EGF-mediated MAPK3/1 on cytoplasmic maturation was examined though in vitro fertilization and embryonic development. The rate of germinal vesicle breakdown (GVBD) after 6 h of culture with 10⁻⁴ mol/l of U0126 (50.4%) was significantly decreased compared with control (67.2%, p < 0.05), and the first polation body (PB1) extrusion rate after 22 h of culture in drug treatment was also significantly inhibited compared with control (28.6% vs. 48.4%, p < 0.05). However, 10-6 mol/l U0126 had slight effect on oocyte nuclear maturation. The normal distribution rate of α-tubulin in the oocytes after 22 h of in vitro maturation was significantly decreased in the 10⁻⁴ mol/l U0126 group (54%) compared with control (68%, p < 0.05). After in vitro fertilization, the cleavage rate in drug treatments (56.8% in 10⁻⁶ mol/l U0126 group and 42.6% in 10⁻⁴ mol/l U0126 group) was significantly decreased compared with control (72.3%, p < 0.01). The blastocyst rate in 10⁻⁴ mol/l U0126 group (17.6%) was also significantly decreased compared with control (29.9%, p < 0.05). Collectively, these results suggest that EGF-mediated MAPK3/1 pathway is conducive to in vitro maturation of sheep oocytes.

Background: Sparganosis caused by invasion of the plerocercoid larvae (spargana) of Spirometra erinaceieuropaei have increased in recent years in China. However, the population genetic structure regarding this parasite is still unclear. In this study, we used the sequences of two mitochondrial genes cytochrome b (cytb) and cytochrome c oxidase subunit I (cox1) to analyze genetic variation and phylogeographic structure of the S. erinaceieuropaei populations., Methodology/principal Findings: A total of 88 S. erinaceieuropaei isolates were collected from naturally infected frogs in 14 geographical locations of China. The complete cytb and cox1 genes of each sample was amplified and sequenced. Total 61 haplotypes were found in these 88 concatenated sequences. Each sampled population and the total population have high haplotype diversity (Hd), accompanied by very low nucleotide diversity (Pi). Phylogenetic analyses of haplotypes revealed two distinct clades (HeN+HuN+GZ-AS clade and GX+HN+GZ-GY clade) corresponding two sub-networks yielded by the median-joining network. Pairwise FST values supported great genetic differentiation between S. erinaceieuropaei populations. Both negative Fu's FS value of neutrality tests and unimodal curve of mismatch distribution analyses supported demographic population expansion in the HeN+HuN+GZ-AS clade. The BEAST analysis showed that the divergence time between the two clades took place in the early Pleistocene (1.16 Myr), and by Bayesian skyline plot (BSP) an expansion occurred after about 0.3 Myr ago., Conclusions: S. erinaceieuropaei from central and southern China has significant phylogeographic structure, and climatic oscillations during glacial periods in the Quaternary may affect the demography and diversification of this species.

High rate of fetal mortality in ruminant somatic cell nuclear transfer (SCNT) pregnancies is due, at least in part, to immune-mediated abortion of fetuses. In the present study, goat uterine leukocytes were isolated by Dolichos biflorus agglutinin (DBA) coated magnetic beads, and with majority being were CD56+CD16- in phenotype with low levels of perforin and Granzyme B expression. The responses of the isolated cells to SCNT and in vitro fertilization (IVF) embryos conditioned mediums containing hormone steroids were compared by measuring their phenotype and cytokines expression. The results showed there was a 2-fold increase in the numbers of isolated uterine leukocytes after incubation with different conditioned mediums for 120 h. However, significantly lower percentage and absolute numbers of uterine CD56+CD16- leukocytes incubated with SCNT conditioned mediums were detected as compared with those incubated with IVF conditioned mediums (P < 0.05). The group treated with progesterone (P4) or the combination of P4 and 17β-estradiol (E2) were associated with significantly higher percentage and absolute numbers of CD56+CD16- cells as compared with those treated with E2 alone (P < 0.05). Furthermore, in the presence of steroids, the isolated leukocytes incubated with SCNT conditioned mediums associated with greater levels of IFN-γ secretion and expression, as well as lesser levels of VEGF, as compared with those treated with IVF conditioned mediums (P < 0.05). In conclusion, this study demonstrates that SCNT embryos have a profound effect on the phenotype expression of goat uterine DBA+ leukocytes, as well as the secretion and expression of IFN-γ and VEGF by these cells in vitro.

Avian leukosis virus subgroup J (ALV-J) has induced serious clinical outbreaks and has become a serious infectious disease of chickens in China. We describe here the creation of a recombinant ALV-J tagged with the enhanced green fluorescent protein (named rHPRS-103EGFP). We successfully utilize the rHPRS-103EGFP to visualize viral infection and for development of a simplified serum-neutralization test.