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Kinship and pedigree, used for estimating inbreeding, heritability, selection, and gene flow, are useful for breeding and animal conservation. However, as the size of crossbred populations increases, inaccurate generation and parentage assignment in livestock farms increase. Restriction-site-associated DNA sequencing is a cost-effective platform for single nucleotide polymorphism (SNP) discovery and genotyping. Here, we performed a kinship analysis and pedigree reconstruction for Angus and Xiangxi yellow cattle. A total of 975 cattle, including 923 offspring with 24 known sires and 28 known dams, were sampled and subjected to SNP discovery and genotyping. The identified SNP panel included 7,305 SNPs capturing the maximum difference between paternal and maternal genome information, allowing us to distinguish F1 from F2 generations with 90% accuracy. In conclusion, we provided a low-cost and efficient SNP panel for kinship analyses and the improvement of local genetic resources, which are valuable for breed improvement, local resource utilization, and conservation.

Collagen is the oldest and most abundant extracellular matrix protein and has many applications in biomedical, food, cosmetic, and other industries. Previous reviews have already introduced collagen’s sources, structures, and biosynthesis. The biological and mechanical properties of collagen-based composite materials, their modification and application forms, and their interactions with host tissues are pinpointed. It is worth noting that self-assembly behavior is the main characteristic of collagen molecules. However, there is currently relatively little review on collagen-based composite materials based on self-assembly. Herein, we briefly reviewed the biosynthesis, extraction, structure, and properties of collagen, systematically presented an overview of the various factors and corresponding characterization techniques that affect the collagen self-assembly process, and summarize and discuss the preparation methods and application progress of collagen-based composite materials in different fields. By combining the self-assembly behavior of collagen with preparation methods of collagen-based composite materials, collagen-based composite materials with various functional reactions can be selectively prepared, and these experiences and outcomes can provide inspiration and practical techniques for the future development directions and challenges of collagen-based composite biomaterials in related applications fields.

Abstract Background Microvascular dysfunction is one of the most common pathological characteristics in Type 2 diabetes. Human mesenchymal stem cell-derived exosomes (hUCMSCs-Exo) have diverse functions in improving microcirculation; however, the molecular mechanism of hUCMSCs-Exo in regulating burn-induced inflammation is not well understood. Methods hUCMSCs-Exo were extracted by hypervelocity centrifugation method, and exosome morphology was observed by transmission electron microscopy, exosome diameter distribution was detected by particle size analysis, and exosome specific proteins were identified by Western blot.2. DB/DB mice were randomly divided into exosomes group and PBS group. Exosomes and PBS were injected into the tail vein, respectively, and the calf muscle tissue was taken 28 days later. 0.5% Evans blue fluorescence assessment microvascular permeability. The expression of CD31 was detected by immunofluorescence.The morphology and function of microvessels in muscle tissue of lower limbs was evaluated by transmission electron microscopy.3. TMT proteomics was used to detect the changes of differential protein expression in lower limb muscle tissues of the PBS group and the exosome group, and data analysis was performed to screen key signal molecules and their involved biological pathways. Key signal molecules CD105 were verified by Western blot. The expression of TGF-β1 in exosomes were evaluated by Western blot. Results Electron microscopy showed that hUCMSCs-Exo presented a uniform vesicle structure, and NTA showed that its diameter was about 160 nm. Western blot showed positive expression of specific proteins CD9, CD81 and TSG101 on exosomes.2. There is no significant change in blood glucose and body weight before and after the exosome treatment. The exosome group can significantly reduce the exudation of Evans blue. Compared with the PBS group. Meanwhile, CD31 immunofluorescence showed that the red fluorescence of exosome treatment was significantly increased, which was higher than that of PBS group. Transmission electron microscopy showed smooth capillary lumen and smooth and complete surface of endothelial cells in the exosome group, while narrow capillary lumen and fingerlike protrusion of endothelial cells in the PBS group.3.Quantitative analysis of TMT proteomics showed that there were 82 differential proteins, including 49 down-regulated proteins and 33 up-regulated proteins. Go enrichment analysis showed that the differential proteins were involved in molecular function, biological process, cell components,among which CD105 was one of the up-regulated proteins. Through literature search, CD105 was found to be related to endothelial cell proliferation. Therefore, this study verified the changes of CD105 in the exosome group, and it was used as the mechanism study of this study. 4. Western blot analysis showed that the expression of CD105 protein in lower limb muscle tissue of exosome group was significantly increased compared with that of PBS group. Based on the fact that CD105 is a component of the TGF-β1 receptor complex and exosomes are rich in growth factors and cytokines, this study further examined the expression of TGF-β1 in exosomes, and the results showed that exosomes had high expression of TGF-β1. Conclusion By improving the integrity of microvascular endothelial cells, hUCMSCs-Exo can improve the permeability of microvessels in diabetic lower muscle tissue, further promote the proliferation of lower limb muscle cells and inhibit the apoptosis of tissue cells. The mechanism may be associated with exosomes rich in TGF-β1, which is likely to promote endothelial cell proliferation and improve permeability through binding to the endothelial CD105/TβR-II receptor complex, while promoting angiogenesis and protecting skeletal muscle cells from apoptosis.

Abstract Background An accurate and reproducible next-generation sequencing platform is essential to identify malignancy-related abnormal DNA methylation changes and translate them into clinical applications including cancer detection, prognosis, and surveillance. However, high-quality DNA methylation sequencing has been challenging because poor sequence diversity of the bisulfite-converted libraries severely impairs sequencing quality and yield. In this study, we tested MGISEQ-2000 Sequencer’s capability of DNA methylation sequencing with a published non-invasive pancreatic cancer detection assay, using NovaSeq6000 as the benchmark. Results We sequenced a series of synthetic cell-free DNA (cfDNA) samples with different tumor fractions and found MGISEQ-2000 yielded data with similar quality as NovaSeq6000. The methylation levels measured by MGISEQ-2000 demonstrated high consistency with NovaSeq6000. Moreover, MGISEQ-2000 showed a comparable analytic sensitivity with NovaSeq6000, suggesting its potential for clinical detection. As to evaluate the clinical performance of MGISEQ-2000, we sequenced 24 clinical samples and predicted the pathology of the samples with a clinical diagnosis model, PDACatch classifier. The clinical model performance of MGISEQ-2000’s data was highly consistent with that of NovaSeq6000’s data, with the area under the curve of 1. We also tested the model’s robustness with MGISEQ-2000’s data when reducing the sequencing depth. The results showed that MGISEQ-2000’s data showed matching robustness of the PDACatch classifier with NovaSeq6000’s data. Conclusions Taken together, MGISEQ-2000 demonstrated similar data quality, consistency of the methylation levels, comparable analytic sensitivity, and matching clinical performance, supporting its application in future non-invasive early cancer detection investigations by detecting distinct methylation patterns of cfDNAs.

The Nordic Sea overflow, being a significant driver of the thermohaline circulation, exerts a substantial influence on environmental dynamics in the Arctic and globally. A better understanding about the trend of the storage volume of the Nordic Seas Overflows is of paramount importance to a realistic assessment of the North Atlantic circulation and variability. EN4.2.2 reanalysis data were utilized to acquire the monthly average time series of overflow water storage volume in the Nordic Sea from 1950 to 2022. The storage volume demonstrates seasonal variations, with fluctuations of approximately 10% around the average. Over the period from 1980 to 2022, the average volume per decade exhibited a consistent decrease. Linear fitting of the annual average data estimated the overall relative change trend in the last 43 years to be -7.2 ± 2.6%. However, the quality of the EN4.2.2 data brings the average error of about 12% in the calculation of the overflow storage volume, which implies that the downward trend requires further validation. From a spatial standpoint, the Norwegian Sea, particularly the Lofoten Basin, is the primary region where overflow water storage volume in the Nordic seas have decreased. This decrease corresponds to an increase in ocean temperatures within the upper layer (0-600m) of both the Lofoten Basin and the Norwegian Basin. The warming of these regions has directly impacted the overflow water storage volume, leading to its reduction. It is noteworthy that the ocean temperature rise in the upper layer is more influenced by the Atlantic inflow rather than air-sea flux, particularly in the Norwegian Basin. This is attributed to the substantial increase in SST in the North Atlantic, which aligns with the warm current regions in the Atlantic Ocean, and the lack of statistical significance in linear trend of the air-sea heat flux. Furthermore, there has been a more rapid reduction in the overflow storage volume in the Norwegian Sea from 2011 to 2022. Specifically, the annual average overflow volume from 2018 to 2022 dropped below the previous lowest value. Concurrently, the salinity of the upper Norwegian Sea and the Atlantic inflow decreased considerably, while there has been no significant change in ocean temperature. Therefore, this short-term fluctuation is predominantly attributed to the decrease in Atlantic inflow salinity. Interestingly, the decrease in overflow water volume in the Nordic Seas does not follow a linear pattern in relation to increasing ocean temperature; instead, it exhibits an accelerating trend. If the ocean temperature rises by 1°C, the overflow water volume in the Nordic Seas will decrease to 86% of the total volume. Overall, the overflow water storage in the Norwegian Sea undergoes complex interannual variations and is notably influenced by the influx from the Atlantic Ocean.

Abstract Objective In order to ensure the stable transcription of target genes, we constructed a eukaryotic high expression vector carrying an immune‐check inhibitor PD‐1v and a variety of cytokines, and studied their effects on activating immune response to inhibit tumor growth. Methods A novel eukaryotic expression plasmid vector named pT7AMPCE containing T7RNA polymerase, T7 promoter, internal ribosome entry site (IRES), and poly A tailing signal was constructed by T4 DNA ligase, on which homologous recombination was used to clone and construct the vector carrying PD‐1v, IL‐2/15, IL‐12, GM‐CSF, and GFP. In vitro transfection of CT26 cells was performed, and the protein expression of PD‐1v, IL‐12 and GM‐CSF was detected by Western blot and ELISA after 48 h. Mice were subcutaneously inoculated with CT26‐IRFP tumor cells in the rib abdomen, and the tumor tissues were injected with PD‐1v, IL‐2/15, IL‐12, and GM‐CSF recombinant plasmids for treatment during the experimental period. The efficacy of the treatment was evaluated by assay tumor size and survival time of tumor‐bearing mice during the experiment. Expression levels of IFN‐γ, TNF, IL‐4, IL‐2, and IL‐5 in mouse blood were measured using the CBA method. Tumor tissues were extracted and immune cell infiltration in tumor tissues was detected by HE staining and the IHC method. Results The recombinant plasmids carrying PD‐1v, IL‐2/15, IL‐12, and GM‐CSF were successfully constructed, and the Western blot and ELISA results showed that PD‐1v, IL‐12, and GM‐CSF were expressed in the supernatant of CT26 cells 48 h after in vitro cell transfection. The combined application of PD‐1v, IL‐2/15, IL‐12, and GM‐CSF recombinant plasmids significantly inhibited tumor growth in mice, and the tumor growth rate was significantly lower than that in the blank control group and GFP plasmid control group (p

The vehicle-road-cloud collaborative V2X system has gradually become a national strategy, and the safety of V2X is related to driving security, life security, property security and even national security, which has increasingly become a hot spot for industry research.Firstly, the overall situation of the V2X security industry and the technology architecture of vehicle-road-cloud collaboration were introduced.Secondly, based on the “vehicle-road-cloud” technology system of V2X, the current status of domestic and international research was discussed, and the remaining problems and challenges in the field of V2X security protection were analyzed from three levels: V2X terminal security, roadside security and cloud security.Finally, the future development and research focus of the V2X security and protection technology were foreseen.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread worldwide and become a major global public health concern. Although novel investigational COVID-19 antiviral candidates such as the Pfizer agent PAXLOVID™, molnupiravir, baricitinib, remdesivir, and favipiravir are currently used to treat patients with COVID-19, there is still a critical need for the development of additional treatments, as the recommended therapeutic options are frequently ineffective against SARS-CoV-2. The efficacy and safety of vaccines remain uncertain, particularly with the emergence of several variants. All 10 versions of the National Health Commission's diagnosis and treatment guidelines for COVID-19 recommend using traditional Chinese medicine. Xuanfei Baidu Decoction (XFBD) is one of the ''three Chinese medicines and three Chinese prescriptions'' recommended for COVID-19. This review summarizes the clinical evidence and potential mechanisms of action of XFBD for COVID-19 treatment. With XFBD, patients with COVID-19 experience improved clinical symptoms, shorter hospital stay, prevention of the progression of their symptoms from mild to moderate and severe symptoms, and reduced mortality in critically ill patients. The mechanisms of action may be associated with its direct antiviral, anti-inflammatory, immunomodulatory, antioxidative, and antimicrobial properties. High-quality clinical and experimental studies are needed to further explore the clinical efficacy and underlying mechanisms of XFBD in COVID-19 treatment.

This paper describes protocols and experiences from a seven-year natural-experiment study in El Paso, Texas, a border city of predominantly Latino/Hispanic population. The study focuses on how Bus Rapid Transit (BRT) impacts physical activity and thus plays a role in alleviating obesity and related chronic diseases that impact healthy aging. Our protocols describe a longitudinal and case-comparison study, which compared residents exposed to new BRT stations with those who were not. This paper also introduces lessons and experiences to overcome the following challenges: delays in the BRT opening (the main intervention), the COVID-19 pandemic, methodological challenges, participant recruitment and retention, and predatory survey takers. Our transdisciplinary approach was pivotal in addressing these challenges. We also proposed and tested multi-level intervention strategies to reduce modifiable barriers to transit use. Our most important takeaway for researchers, practitioners, and policy makers is the importance of being flexible and ready to adapt to new circumstances. Future natural-experiment researchers need to become more versatile in an increasingly volatile and uncertain world.

Summary: Background: Cell-free DNA (cfDNA) is being explored as biomarker for non-invasive diagnosis of cancer. We aimed to establish a cfDNA-based DNA methylation marker panel to differentially diagnose papillary thyroid carcinoma (PTC) from benign thyroid nodule (BTN). Methods: 220 PTC- and 188 BTN patients were enrolled. Methylation markers of PTC were identified from patients’ tissue and plasma by reduced representation bisulfite sequencing and methylation haplotype analyses. They were combined with PTC markers from literatures and were tested on additional PTC and BTN samples to verify PTC-detecting ability using targeted methylation sequencing. Top markers were developed into ThyMet and were tested in 113 PTC and 88 BTN cases to train and validate a PTC-plasma classifier. Integration of ThyMet and thyroid ultrasonography was explored to improve accuracy. Findings: From 859 potential PTC plasma-discriminating markers that include 81 markers identified by us, the top 98 most PTC plasma-discriminating markers were selected for ThyMet. A 6-marker ThyMet classifier for PTC plasma was trained. In validation it achieved an Area Under the Curve (AUC) of 0.828, similar to thyroid ultrasonography (0.833) but at higher specificity (0.722 and 0.625 for ThyMet and ultrasonography, respectively). A combinatorial classifier by them, ThyMet-US, improved AUC to 0.923 (sensitivity = 0.957, specificity = 0.708). Interpretation: The ThyMet classifier improved the specificity of differentiating PTC from BTN over ultrasonography. The combinatorial ThyMet-US classifier may be effective in preoperative diagnosis of PTC. Funding: This work was supported by the grants from National Natural Science Foundation of China (82072956 and 81772850).

Protein aggregation and associated amyloid formation is linked with several harmful human pathophysiologies including Alzheimer's, Parkinson's, and cerebrovascular diseases. A potential approach for modulating and exploring amyloid fibrillization is the control of protein self-assembly. Herein, anti-aggregation effects of salidroside, its influence on the kinetics of amyloid fibrillization of Aβ1-42 peptide and its cytotoxicity against cerebrovascular endothelial cells (bEnd.3) were assessed by using a wide range of spectroscopic and cellular techniques. The present outcome of Thioflavin T (ThT) and 8-anilino-1-naphthalenesulfonic acid (ANS) fluorescence, Congo red (CR), and circular dichroism (CD) analyses indicated that salidroside potentially inhibits protein fibril formation. The cellular studies inferred that salidroside protects bEnd.3 cells against Aβ1-42 oligomers -triggered cytotoxicity through modulation of oxidative stress [reactive oxygen species (ROS), superoxide dismutase (SOD) and catalase (CAT) activities] and apoptosis (caspase-3 activity). Therefore, the data signifies the role of salidroside as a promising small molecule in inhibiting Aβ1-42 aggregation and associated cerebrovascular endothelial cell toxicity. Hence, salidroside can serve as a potential inhibitor in the therapeutic advancement to combat cerebrovascular diseases.

Near-surface PM2.5 estimates remain a global scientific research challenge due to their effect on human fitness and atmospheric environmental quality. However, practical near-surface PM2.5 estimates are impeded by the incomplete monitoring data. In this study, we propose the trigonometric polynomial fitting (TPF) method to estimate near-surface PM2.5 concentrations in south-central China during 2015. We employ 10-fold cross-validation (CV) to assess the reliability of TPF in estimating practical PM2.5 values. When compared to alternative methods such as the orthogonal polynomial fitting (OBF) method based on Chebyshev basis functions, Kriging interpolation, and radial basis function (RBF) interpolation, our results show that utilizing TPF31, with a maximum order of 3 in the x direction and a maximum order of 1 in the y direction, leads to superior efficiency through error minimization. TPF31 reduces MAE and RMSE by 1.93%, 24%, 6.96% and 3.6%, 23.07%, 10.43%, respectively, compared to the other three methods. In addition, the TPF31 method effectively reconstructs the spatial distribution of PM2.5 concentrations in the unevenly distributed observation stations of Inner Mongolia and the marginal regions of the study area. The reconstructed spatial distribution is remarkably smooth. Despite the non-uniform distribution of observation stations and the presence of missing data, the TPF31 method demonstrates exceptional effectiveness in accurately capturing the inherent physical attributes of spatial distribution. The theoretical and experimental results emphasize that the TPF method holds significant potential for accurately reconstructing the spatial distribution of PM2.5 in China.

Based on the method for identifying the boundary of movable water bodies (MWBB), the spatial distribution of reclamation projects in the Guangxi Beibu Gulf were identified over the past 40 years and the impact of these engineering facilities on hydrodynamics was also evaluated. The results showed that 163.8 km2 of natural sea areas in the Guangxi Beibu Gulf were occupied through reclamation in the last 40 years. The effects of land reclamation on tidal amplitude were more pronounced in the second period (2001–2018) than in the first period (1987–2001), particularly in the tidal channels of Qinzhou Bay and Fangcheng Bay, where the amplitude difference ranged from 8 to 15 cm, representing a 40–55% increase. The reduction in the sea area because of land reclamation has changed the hydrodynamics in the Guangxi Beibu Gulf, including reducing the tidal volume, altering the amplitude variations, and increasing the seaward residual currents, all of which could cause significant problems for the coastal environment.

Citrullinated neoepitopes have emerged as key triggers of autoantibodies anti-citrullinated protein antibodies (ACPA) synthesis in rheumatoid arthritis (RA) patients. Apart from their critical role in homeostasis and thrombosis, platelets have a significant contribution to inflammation as well. Although anuclear in nature, platelets have an intricate post-translational modification machinery. Till now, citrullination in platelets and its contribution to trigger autoantibodies ACPA production in RA is an unexplored research direction. Herein, we investigated the expression of peptidylarginine deiminase (PAD) enzymes and citrullinated proteins/peptides in the human platelets and platelet derived microparticles (PDP). Both PAD4 mRNA and protein, but not the other PAD isoforms, are detectable in the human platelets. With a strict filtering criterion,108 citrullination sites present on 76 proteins were identified in the human platelets, and 55 citrullinated modifications present on 37 different proteins were detected in the PDPs. Among them, some are well-known citrullinated autoantigens associated with RA. Citrullinated forms of thrombospondin-1, β-actin, and platelet factor-4 (also known as CXCL4) are highly immunogenic and bound by autoantibodies ACPA. Furthermore, ACPA from RA sera and synovial fluids recognized citrullinated proteins from platelets and significantly activated them as evidenced by P-selectin upregulation and sCD40 L secretion. These results clearly demonstrate the presence of citrullinated autoantigens in platelets and PDPs, thus could serve as potential targets of ACPA in RA.

The origin and progression of lung adenocarcinoma (LUAD) are still poorly understood. Here, the authors analyse LUAD composition and progression in patient samples using single-cell RNA-seq and multiplex imaging, revealing a potential transcriptional divergence from alveolar type 2 cells.

Abstract Purpose Multiple anthropogenic disturbances, such as climate warming and nitrogen deposition are affecting terrestrial ecosystems. Different disturbances may have some consistent effects on the soil microbial community, which remains largely unexplored. Methods We mimicked 16 anthropogenic disturbances in a steppe ecosystem, and measured the absolute abundance and taxonomic composition of soil bacterial communities with qPCR and amplicon sequencing, respectively. Results We found that while the absolute abundance of each of the four dominant bacterial phyla did not show a consistent response to these disturbances, that of the five subdominant phyla showed a consistent increase. Meanwhile, these disturbances consistently stimulated the relative abundances of metabolic functions for high-growth-yield, including the transport/metabolism of amino acids and carbohydrates. Stochastic processes (e.g., random birth) played more critical roles in structuring the subdominant than dominant phyla, and the disturbances promoted the stochastic processes. Conclusions Overall, the high-yield traits and stochasticity of subdominant phyla led to their positive responses to disturbances. Furthermore, our findings indicate that the intensifying human activities are likely to cause a high-yield-strategies-toward shift in soil microbial composition in the Eurasian steppe ecosystem.

IntroductionPost-stroke aphasia (PSA) is a language disorder caused by left hemisphere stroke. Electroacupuncture (EA) is a minimally invasive therapeutic option for PSA treatment. Tongli (HT5) and Xuanzhong (GB39), two important language-associated acupoints, are frequently used in the rehabilitation of patients with PSA. Preliminary evidence indicated functional activation in distributed cortical areas upon HT5 and GB39 stimulation. However, research on the modulation of dynamic and static functional connectivity (FC) in the brain by EA in PSA is lacking.MethodThis study aimed to investigate the PSA-related effects of EA stimulation at HT5 and GB39 on neural processing. Thirty-five participants were recruited, including 19 patients with PSA and 16 healthy controls (HCs). The BOLD signal was analyzed by static independent component analysis, generalized psychophysiological interactions, and dynamic independent component analysis, considering variables such as age, sex, and years of education.ResultsThe results revealed that PSA showed activated clusters in the left putamen, left postcentral gyrus (PostCG), and left angular gyrus in the salience network (SN) compared to the HC group. The interaction effect on temporal properties of networks showed higher variability of SN (F = 2.23, positive false discovery rate [pFDR] = 0.017). The interaction effect on static FC showed increased functional coupling between the right calcarine and right lingual gyrus (F = 3.16, pFDR = 0.043). For the dynamic FC, at the region level, the interaction effect showed lower variability and higher frequencies of circuit 3, with the strongest connections between the supramarginal gyrus and posterior cingulum (F = 5.42, pFDR = 0.03), middle cingulum and PostCG (F = 5.27, pFDR = 0.036), and triangle inferior frontal and lingual gyrus (F = 5.57, pFDR = 0.026). At the network level, the interaction effect showed higher variability in occipital network–language network (LN) and cerebellar network (CN) coupling, with stronger connections between the LN and CN (F = 4.29, pFDR = 0.042). Dynamic FC values between the triangle inferior frontal and lingual gyri were anticorrelated with transcribing, describing, and dictating scores in the Chinese Rehabilitation Research Center for Chinese Standard Aphasia Examination.DiscussionThese findings suggest that EA stimulation may improve language function, as it significantly modulated the nodes of regions/networks involved in the LN, SN, CN, occipital cortex, somatosensory regions, and cerebral limbic system.

The ecosystem parameters are critical for precisely determining the marine ecological process and improving the simulations of the marine ecological model. In this study, based on the NPZD (nutrient, phytoplankton, zooplankton and detritus) model, the surface chlorophyll-a observations obtained from Sea-viewing Wide Field-of-view Sensor (SeaWiFS) data were assimilated to estimate spatially ecosystem parameters in the Bohai, Yellow, and East China Seas using an adjoint assimilation method with characteristic finite difference scheme. The experiments of the moving Gaussian hump indicated that the characteristic finite difference method (CFDM) can get rid of the limit of stability and permit using large time steps, which reduces long computation durations and large memory requirements. The model performance was significantly improved after data assimilation with CFDM using a large time step of 6 hours. Moreover, the distributions of parameters of the NPZD model in winter in the Bohai Sea, the Yellow Sea, and the East China Sea were simulated by our method. Overall, the developed method can efficiently optimize the ecosystem parameters and the results can be beneficial for determining reasonable parameters of the marine ecological model.

The vertical eddy viscosity coefficient (VEVC) is an important parameter used in ocean dynamics studies to describe the intensity of mixing in the vertical direction and the process of momentum transport. In this paper, an adjoint assimilation method was used to invert the VEVC, based on the Ekman model, with the measured wind field and current data. The main purpose was to study the effect of changes in the background wind field on VEVC, and thus investigate the role of wind stress in the inversion process. The results indicated that the inverse vertical eddy viscosity coefficient increased slightly at the surface layer, reaching its maximum at around 10–12 m, and then decreased monotonically with depth. The maximum VEVC value corresponds to different depths for different wind speed ranges. Additionally, wind steering could affect the VEVC inversion curve, causing it to deviate from the general trend. The kinetic energy ratio increased with depth, peaked at 18–20 m, and then rapidly decreased to nearly zero beyond 24 m. The impact of wind field strength and steering on VEVC was observed in the kinetic energy ratio curve, which confirms the speculative wind stress effect. This study revealed the characteristics and mechanisms of VEVC in coastal waters under different wind conditions, which could provide a reference for further research on physical oceanography.

Abstract Background Stroke‐associated pneumonia (SAP) occurs frequently after a stroke. Geriatric Nutritional Risk Index (GNRI) is a valuable indicator of elderly individuals’ nutritional status. This research was designed to obtain insight into the link between GNRI and SAP. Methods Patients with acute ischemic stroke (AIS) were categorized into the SAP and non‐SAP groups. GNRI scores were divided into four layers: Q1, GNRI 98. To identify the independent risk and protective factors of developing SAP, logistic regression analyses were conducted. Additionally, we utilized the restricted cubic spline (RCS) analysis to test the effect of GNRI on the SAP risk. Results The SAP group showed lower GNRI scores than the non‐SAP group (96.88 ± 9.36 vs. 100.88 ± 8.25, p

BackgroundIt has been proven that the ratio of albumin to globulin (A/G) is a typical biomarker for monitoring inflammation and nutritional status. But the potential role of A/G in stroke-associated pneumonia (SAP) after acute ischemic stroke (AIS) remained unknown.Patients and methodsFollowing inclusion criteria, 5,173 AIS patients were included and segmented into SAP (n = 897) and non-SAP (n = 4,276) groups. The differences in variables between groups were compared. The logistic regression model was used to determine the association between A/G and SAP, and a forest plot was drawn.ResultsCompared with the non-SAP group, the SAP group had a lower A/G level (P < 0.001). Then, A/G was divided into quartiles. In comparison to Q3 (A/G = 1.25–1.39), logistic regression revealed that patients with a lower A/G (A/G ≤ 1.09) had a higher risk of SAP (OR = 1.96, 95% CI, 1.56–2.46, P < 0.001). On the contrary, those with a higher A/G (A/G ≥ 1.4) had a lower SAP risk (OR = 0.73, 95% CI, 0.54–0.97, P = 0.029).ConclusionThe study revealed that a low A/G level was associated with an increased SAP risk. Appropriate preventative measures for SAP should be taken in AIS patients with a low A/G level.

The harmonic constants of long-period tidal constituents are critical for simulating the tide process, obtaining theoretical datum sounding reduction, and conducting further ocean research. In this paper, the equidistant node orthogonal polynomial fitting (ENOPF) method is employed to fit the harmonic constants of the T/P satellite altimeter data and the gridded altimeter data from the Data Unification and Altimeter Combination System (DUACS) to obtain the full-field harmonic constants of the long-period tidal constituents (Sa and Ssa) in the Bohai Sea, Yellow Sea, and East China Sea (BYECS). To verify the validity of the ENOPF method, the long-period tidal harmonic constants of the ENOPF method, the Finite Element Solutions 2014 (FES2014) model, the Empirical Ocean Tide 20 (EOT20) model, and the DUACS data were compared with the observations of the X-TRACK products and the tide gauges. In addition, the root-mean-square errors (RMSEs), amplitude differences (ΔH), and phase-lag differences (ΔG) of the comparison results were calculated. According to the error analysis and the cotidal charts, the ENOPF method is better than the other three methods or models for comparing tide gauges (6.19 cm, 5.85 cm, 25.44°; 1.10 cm, 0.72 cm, 59.09°, respectively. The cotidal charts obtained by the ENOPF method are smoother and have better consistency with the actual track data. The results indicate that, due to the polynomial fitting method adopted in ENOPF method, which has the characteristics of improving the resolution infinitely, it is easier to obtain the harmonic constant of full field than with other models or methods. This study proves that the ENOPF method is a reasonable and simple tool in extracting the harmonic constants of the BYECS.

By reviewing the development history of stimulation techniques for deep/ultra-deep oil and gas reservoirs, the new progress in this field in China and abroad has been summed up, including deeper understanding on formation mechanisms of fracture network in deep/ultra-deep oil and gas reservoir, performance improvement of fracturing fluid materials, fine stratification of ultra-deep vertical wells, and mature staged multi-cluster fracturing technique for ultra-deep and highly deviated wells/horizontal wells. In light of the exploration and development trend of ultra-deep oil and gas reservoirs in China, the requirements and technical difficulties in ultra-deep oil and gas reservoir stimulation are discussed: (1) The research and application of integrated geological engineering technology is difficult. (2) The requirements on fracturing materials for stimulation are high. (3) It is difficult to further improve the production in vertical profile of the ultra-deep and hugely thick reservoirs. (4) The requirements on tools and supporting high-pressure equipment on the ground for stimulation are high. (5) It is difficult to achieve efficient stimulation of ultra-deep, high-temperature and high-pressure wells. (6) It is difficult to monitor directly the reservoir stimulation and evaluate the stimulation effect accurately after stimulation. In line with the complex geological characteristics of ultra-deep oil and gas reservoirs in China, seven technical development directions are proposed: (1) To establish systematic new techniques for basic research and evaluation experiments; (2) to strengthen geological research and improve the operational mechanism of integrating geological research and engineering operation; (3) to develop high-efficiency fracturing materials for ultra-deep reservoirs; (4) to research separated layer fracturing technology for ultra-deep and hugely thick reservoirs; (5) to explore fracture-control stimulation technology for ultra-deep horizontal well; (6) to develop direct monitoring technology for hydraulic fractures in ultra-deep oil and gas reservoirs; (7) to develop downhole fracturing tools with high temperature and high pressure tolerance and supporting wellhead equipment able to withstand high pressure.

BackgroundStakeholders from multiple sectors are increasingly aware of the critical need for identifying sustainable interventions that promote healthy lifestyle behaviors. Activity-friendly communities (AFCs) have been known to provide opportunities for engaging in physical activity (PA) across the life course, which is a key to healthy living and healthy aging.PurposeOur purpose is to describe the study protocol developed for a research project that examines: (a) the short- and long-term changes in total levels and spatial and temporal patterns of PA after individuals move from non-AFCs to an AFC; and (b) what built and natural environmental factors lead to changes in PA resulting from such a move, either directly or indirectly (e.g., by affecting psychosocial factors related to PA).MethodsThis protocol is for a longitudinal, case-comparison study utilizing a unique natural experiment opportunity in Austin, Texas, USA. Case participants were those adults who moved from non-AFCs to an AFC. Matching comparison participants were residents from similar non-AFCs who did not move during the study period. Recruitment venues included local businesses, social and print media, community events, and individual referrals. Objectively measured moderate-to-vigorous PA and associated spatial and temporal patterns served as the key outcomes of interest. Independent (e.g., physical environments), confounding (e.g., demographic factors), and mediating variables (e.g., psychosocial factors) were captured using a combination of objective (e.g., GIS, GPS, Tanita scale) and subjective measures (e.g., survey, travel diary). Statistical analyses will be conducted using multiple methods, including difference-in-differences models, repeated-measures linear mixed models, hierarchical marked space-time Poisson point pattern analysis, and hierarchical linear mixed models.ConclusionNatural experiment studies help investigate causal relationships between health and place. However, multiple challenges associated with participant recruitment, extensive and extended data collection activities, and unpredictable intervention schedules have discouraged many researchers from implementing such studies in community-based populations. This detailed study protocol will inform the execution of future studies to explore how AFCs impact population health across the life course.

Summary: Precipitation may increase or decrease by different intensities, but the pattern and mechanism of soil microbial community assembly under various precipitation changes remain relatively underexplored. Here, although ±30% precipitation caused a small decrease (∼19%) in the within-treatment taxonomic compositional dissimilarity through the deterministic competitive exclusion process in a steppe ecosystem, ±60% precipitation caused a large increase (∼35%) in the dissimilarity through the stochastic ecological drift process (random birth/death), which was in contrast with the traditional thought that increasing the magnitude of environmental changes (e.g., from +30% to +60%) would elevate the importance of deterministic relative to stochastic processes. The increased taxonomic dissimilarity/stochasticity under ±60% precipitation translated into functional dissimilarity/stochasticity at the gene, protein, and enzyme levels. Overall, our results revealed the distinctive pattern and mechanism of precipitation changes affecting soil microbial community assembly and demonstrated the need to integrate microbial taxonomic information to better predict their functional responses to precipitation changes.

BackgroundThe coronavirus disease 2019 (COVID-19) pandemic is having a dramatic impact on acute stroke care. Its effects may accompany stroke care for a long time. We compared the treatment, short-term and long-term functional outcomes of patients with AIS from 2019 to 2020. Our objective was to evaluate the effect of COVID-19 epidemic on mechanical thrombectomy (MT) in patients in our hospital.MethodsWe collected information on subjects treated with MT in 2019–2020, including age, sex, time from the onset to arterial sheath insertion, time from the onset to recanalization, the rate of lung infection and hemorrhagic transformation, modified Rankin scale (mRS), NHISS, and ASPECTS.ResultsThe number of patients with MT decreased significantly by 26.6% in 2020 (p = 0.025). The pretreatment ASPECTS score for 2020 was significantly higher than 2019 (p = 0.004). Besides, the patients were more likely to develop lung infection (65 vs. 54.1%, p = 0.042) and had a higher risk of hemorrhagic transformation (47.4% vs. 30.4%, p = 0.005) in 2019. The discharged mRS reflected the worse short-term functional prognosis of patients with MT in 2019 (66 vs. 44.9%, p = 0.046). In the subgroup analysis of bridging thrombolysis (BT), more patients with BT are expected to have a poor short-term functional prognosis in 2020, according to the discharged mRS (62.5 vs. 37.5%, p = 0.024). However, there was no difference in mRS at 180 days between the two groups (p = 0.094).ConclusionFor patients with MT, both short- and long-term functional outcomes were not significantly affected due to the mild condition of patients admitted to hospital in 2020. For patients with BT, the COVID-19 pandemic has prolonged the green channel time of stroke, leading to a poor short-term functional prognosis of patients with stroke in the pandemic period. There was no difference in the effectiveness of direct MT and BT during the COVID-19 pandemic.

BackgroundPost-stroke fatigue (PSF) is a frequent complication of stroke. Serum uric acid (SUA) is frequently thought to be a risk factor for stroke. This study aimed to investigate whether SUA also played a role in PSF.MethodsSubjects with ischemic stroke were screened from The First Affiliated Hospital of Wenzhou Medical University between January 2020 and October 2020. Patients' fatigue symptoms were assessed by the Fatigue severity scale (FSS). To investigate the relationship between SUA and PSF, binary logistic regression analysis was conducted, with the confounders being controlled. SUA levels were divided into four layers (Q1 ≤ 245 μmol/L; Q2 246–308 μmol/L; Q3 309–365 μmol/L; Q4 ≥366 μmol/L) based on the quartiles.ResultsSUA levels were significantly higher in the PSF group (345.96 ± 73.78 μmol/L) than the non-PSF group (295.97 ± 87.8 μmol/L, P < 0.001). There were no differences in any other variables between these two groups. After adjusting the confounders, the risk of PSF in the Q4 layer (≥366 μmol/L) was 6.05 times (95% CI 1.79–20.43, P = 0.004) higher than that in Q1 (≤245 μmol/L).ConclusionHigh SUA at admission was an independent risk factor for fatigue 1 year after stroke onset. High SUA (≥366 μmol/L) during stroke deserves more attention, and active control of high SUA levels may be beneficial to reduce the incidence of PSF in the chronic stage following stroke.

Objectives. To investigate the effect of placenta-derived mesenchymal stem cells (PMSCs) on diabetic peripheral neuropathy and explore the role of Wnt signaling pathway. Method. Twenty-seven male db/db mice were randomly categorized into the control group, PMSC group, and PMSC treatment with Wnt inhibitor treatment group. Intervention was initiated in week 22. Thermal stimulation response was determined with a plantar analgesia tester. The mice were sacrificed on 7, 14, and 28 days. The morphology of sciatic nerves was observed by electron microscopy, and the expression of protein gene product (PGP) 9.5, S100β, and Ku80 was detected by immunofluorescence. Bax, β-catenin, and dishevelled1 (DVL1) were detected by western blot. Results. Thermal stimulation response was improved in the PMSC group on 14 and 28 days. Compared with the control group, PGP9.5 was increased in the PMSC group, accompanied by a significant increase in the expression of S100β. On the contrary, LGK974 inhibited the effect of PMSCs on thermal stimulation response and the expression of PGP9.5 and S100β. Both PGP9.5 and S100β were correlated with Ku80 in fluorescence colocalization. The myelin sheath of sciatic nerves in the PMSC group was uniform and dense compared with that in the control group. The effects of PMSCs promoting myelin repair were significantly inhibited in the PMSC+LGK974 group. Bax in the PMSC group expressed less than the control group. In contrast, the expressions of β-catenin and DVL1 were higher compared with that in the control group on the 14th and 28th days. The expression of DVL1 and β-catenin was lower in the PMSC+LGK974 group than in the PMSC group. Conclusions. PMSCs improved the symptoms of diabetic peripheral neuropathy, along with the improvement of nerve myelin lesions, promotion of nerve regeneration, and activation of Schwann cells, which might be related to the regulation of Wnt signaling pathway and inhibition of apoptosis.

Background: Stroke-associated pneumonia (SAP) is associated with poor prognosis after acute ischemic stroke (AIS).Purpose: This study aimed to describe the parameters of coagulation function and evaluate the association between the fibrinogen-to-albumin ratio (FAR) and SAP in patients with AIS.Patients and methods: A total of 932 consecutive patients with AIS were included. Coagulation parameters were measured at admission. All patients were classified into two groups according to the optimal cutoff FAR point at which the sum of the specificity and sensitivity was highest. Propensity score matching (PSM) was performed to balance potential confounding factors. Univariate and multivariate logistic regression analyses were applied to identify predictors of SAP.Results: A total of 100 (10.7%) patients were diagnosed with SAP. The data showed that fibrinogen, FAR, and D-dimer, prothrombin time (PT), activated partial thromboplastin time (aPTT) were higher in patients with SAP, while albumin was much lower. Patients with SAP showed a significantly increased FAR when compared with non-SAP (P < 0.001). Patients were assigned to groups of high FAR (≥0.0977) and low FAR (

Abstract Background Primary periodic paralysis is characterized by recurrent quadriplegia typically associated with abnormal serum potassium levels. The molecular diagnosis of primary PP previously based on Sanger sequencing of hot spots or exon-by-exon screening of the reported genes. Methods We developed a gene panel that includes 10 ion channel-related genes and 245 muscular dystrophy- and myopathy-related genes and used this panel to diagnose 60 patients with primary periodic paralysis and identify the disease-causing or risk-associated gene mutations. Results Mutations of 5 genes were discovered in 39 patients (65.0%). SCN4A, KCNJ2 and CACNA1S variants accounted for 92.5% of the patients with a genetic diagnosis. Conclusions Targeted next-generation sequencing offers a cost-effective approach to expand the genotypes of primary periodic paralysis. A clearer genetic profile enables the prevention of paralysis attacks, avoidance of triggers and the monitoring of complications.

High-precision tidal harmonic constants are necessary for studies involving tides. This study proposes a new method combined with the adjoint assimilation model and the Chebyshev polynomial fitting (CPF) method to obtain the tidal harmonic constants in the shallow-water region of the Bohai and Yellow Sea (BYS). Based on the CPF method, the full-field harmonic constants and reliable cotidal charts of the eight major constituents (M2, S2, K1, O1, N2, K2, P1 and Q1) were fitted from the X-TRACK products briefly and this method was effectively for coastal conditions. Compared with the observations of the X-TRACK products and tidal gauges, for the M2 constituent, the TPXO9, Finite Element Solutions 2014 (FES2014), National Astronomical Observatory 99b (NAO.99b) and Empirical Ocean Tide 20 (EOT20) models yield the root-mean-square errors (RMSEs) of 18.50, 7.31, 18.73 and 13.32 cm, respectively, while the CPF method yields an RMSE of 10.74 cm. These results indicate that the CPF method could maintain high resolution and obtain accurate cotidal charts consistent with the simulations of the four models in shallow-water regions.

In this study, a triangular polynomial interpolation (TPI) scheme was developed to estimate the vertical eddy viscosity coefficient (VEVC) on the basis of the Ekman model with adjoint assimilation. In the twin experiments, the advantages and disadvantages of estimating the VEVC using the TPI scheme under different factors are discussed. The results indicated that (1) the TPI scheme proves to be better than the cubic spline interpolation (CSI) and Cressman interpolation (CI) schemes; (2) the inversion results are more sensitive to observations from upper ocean layers than those from lower layers, and the TPI scheme is less likely to be influenced by missing data; (3) for various boundary layer depths, the inversion results of the TPI scheme remain consistent with the given distributions; (4) the inversion results can be influenced considerably by observational errors, and the TPI scheme is more resistant to noise than the CSI and CI schemes; and (5) the inversion accuracy of the TPI scheme can be improved by selecting the temporal wind stress drag coefficients. In practical experiments, the adjoint method with the TPI scheme was developed to estimate the Ekman currents by assimilating the observations from a buoy stationed in the Yellow Sea. The results showed the successful estimation of the VEVC and demonstrated that more precise current velocities can be obtained with this estimation scheme. In summary, this study provides a useful approach for the effective estimation of the VEVC.

Background and Purpose: The results regarding the independent association between homocysteine (Hcy) levels and post-stroke cognitive impairment (PSCI) were inconsistent. The effect of age on this association has yet to be explored. This study aims to determine the relationship between Hcy levels, age, and cognitive impairment in a post-stroke population.Methods: A total of 592 patients with acute ischemic stroke (AIS) completed follow-up. Serum Hcy levels were measured enzymatically by spectrophotometry within 24 h of admission. Cognitive function was evaluated by the Mini-Mental State Examination (MMSE) 1 month after stroke, and the scores ≤ 24 were considered as cognitive impairment. Our study was dichotomized into two groups by a cut-off of 65 years. Multivariate logistic regression models were used to determine the association between baseline Hcy levels and cognitive impairment.Results: According to the MMSE score, 317 (53.5%) patients had cognitive impairment. Patients with higher levels of Hcy were more prone to have cognitive impairment 1 month after stroke than patients with lower levels of Hcy (p < 0.001). The optimal cut-off points of Hcy level (μmol/L) were (T1) ≤ 8, (T2) 8–12, and (T3) ≥ 12. After adjusting for confounding factors, the multivariate regression analysis showed that the third Hcy tertile was independently associated with cognitive impairment [odds ratio (OR) = 2.057, 95% confidence interval (CI) = 1.133–3.735, p = 0.018). A stronger association [T2 (OR = 2.266, 95% CI = 1.042–4.926, p = 0.039); T3 (OR =3.583, 95% CI = 1.456–8.818, p = 0.005)] was found in the younger group. However, the independent association was not confirmed in the older group.Conclusions: Elevated Hcy levels in the acute phase of ischemic stroke were independently associated with cognitive impairment in a post-stroke population. Furthermore, the association was age-dependent and more meaningful in a younger population aged below 65. So, Hcy levels in patients with stroke should be well-monitored, especially in younger patients.

Random migration often plays important roles in soil microbial community assembly. However, it remains unexplored that whether its role is affected by anthropogenic disturbances, and whether the effects of different disturbances are consistent. Here, we report a novel and general pattern that different disturbances consistently restrain the role of migration in soil bacterial community assembly. We mimicked 16 types of anthropogenic disturbances relevant to a Eurasian steppe ecosystem, including removing different number of plant functional groups, mowing, adding nitrogen, adding phosphorus, watering, warming, and some of their combinations. The estimated migration rates in soil bacterial community assembly with Stephen Hubbell’s neutral model decreased by 0.19–6.29% under these disturbances. The relative change in migration rate was correlated with the deterministic change in community composition and the relative change in soil pH. Our results suggested that the intensifying anthropogenic disturbances may restrain the random dispersal, colonization and distribution of soil microorganisms.

Increasing atmospheric nitrogen (N) deposition causes profound changes in the plant community, significantly threatening the terrestrial ecosystem function and stability worldwide. Random migration is a primary determinant of plant community assembly in the neutral model. However, whether its role is affected by N deposition is obscure. Therefore, the effects of eight N application rates (0, 1, 2, 3, 5, 10, 15, and 20 g N m−2 yr−1) on the estimated migration rate of plant community were examined in a long-term field study in a semiarid steppe. Our results indicated that N addition significantly impacted the estimated migration rate (F = 4.89, P

Abstract Background Sutherlandia frutescens is one of the most promising commercialized, indigenous and medicinal plants of South Africa that is used as an immune-booster, and a traditional treatment for cancer. However, few studies report on its toxicology and dosage in vivo. There is still room to better understand its cytotoxicity effects in animal systems. Methods We prepared two extracts, one with 80% (v/v) ethanol, and the other, with water. Both were studied to determine the maximum tolerable concentration when extracts were applied at 0 to 200 μg/ml to a Tuebingen zebrafish embryo line. The development of zebrafish embryos after 24 h post fertilization (hpf) was studied. A concentration range of 5 μg/ml to 50 μg/ml was then chosen to monitor the ontological development of cultured embryos. A liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method was used to study the differences of the two experimental extracts. Chemical variation between the extracts was illustrated using chemometrics. Results Both extracts led to bleeding and pericardial cyst formation when applied at high concentrations to the zebrafish embryo culture. Chronic teratogenic toxicities, leading to pericardial edema, yolk sac swelling, and other abnormal developmental characteristics, were detected. The aqueous extracts of S. frutescens were less toxic to the larvae than the ethanol extracts, validating preference for aqueous preparations when used in traditional medicine. Chemical differences between the water extracts and alcoholic extracts were analysed using LC-MS/MS. A supervised metabolomics approach, targeting the sutherlandiosides and sutherlandins using orthogonal partial least squares-discriminant analysis (OPLS-DA), illustrated that sutherlandiosides were the main chemical features that can be used to distinguish between the two extracts, despite the extracts being highly similar in their chemical constituents. Conclusion The water extract caused less cytotoxic and abnormal developmental effects compared to the ethanolic extract, and, this is likely due to differences in concentrations of extracted chemicals rather than the chemical profile per se. This study provides more evidence of cytotoxicity effects linked to S. frutescens using the zebrafish embryo bioassay as a study tool.

The independent point scheme (IPS) is applied to inverting initial condition with the adjoint method for the ocean pollutant transport model in this work. As an improvement, the linear Cressman interpolation is removed and the surface spline interpolation is implemented in the IPS. A series of numerical experiments are carried out to test and compare the improved IPS. And experiment results show that through applying the improved IPS, what is further reduced is mean absolute errors between simulation results and observations. Moreover, the inverted distributions are more smooth, accurate and reasonable. In addition, the application of improved IPS also reduces the variables that need to be inverted and promotes the computational efficiency. By these numerical experiment results, it is demonstrated that the combination of improved IPS and adjoint method can be used for the inversion of initial conditions and parameters estimation more effectively and reliably.

In this study, the time-varying wind stress drag coefficient in the Ekman model was inverted by the cubic spline interpolation scheme based on the adjoint method. Twin experiments were carried out to investigate the influences of several factors on inversion results, and the conclusions were (1) the inverted distributions with the cubic spline interpolation scheme were in good agreement with the prescribed distributions of the wind stress drag coefficients, and the cubic spline interpolation scheme was superior to direct inversion by the model scheme and Cressman interpolation scheme; (2) the cubic spline interpolation scheme was more advantageous than the Cressman interpolation scheme even if there is moderate noise in the observations. The cubic spline interpolation scheme was further validated in practical experiments where Ekman currents and wind speed derived from mooring data of ocean station Papa were assimilated. The results demonstrated that the variation of the time-varying wind stress drag coefficient with time was similar to that of wind speed with time, and a more accurate inversion result could be obtained by the cubic spline interpolation scheme employing appropriate independent points. Overall, this study provides a potential way for efficient estimation of time-varying wind stress drag coefficient.

Melatonin has been identified in a variety of invertebrate species, but its function is not as well understood as in crustaceans. The effects of melatonin on hemolymph glucose levels and tissue carbohydrate metabolism in the Chinese mitten crab, Eriocheir sinensis, were fully investigated in this study. Moreover, whether the eyestalk (an important endocrine center in invertebrate species) involves in this process or not, also were clarified. Analysis revealed that eyestalk ablation, especially bilateral, caused a significant decrease in the hemolymph glucose level. Moreover, injection of melatonin induced hyperglycemia in a dose-dependent manner both in intact and ablated crabs. Based on the expression of CHH mRNA in the 10 different tissues, eyestalk, thoracic ganglion, intestinal tract and hemolymph were selected to estimate the effect of melatonin on the expression of CHH mRNA. Bilateral eyestalk ablation caused a significant increase in the expression of CHH mRNA in the thoracic ganglion, intestinal tract and hemolymph compared with the controls. In addition, injection of melatonin into intact or ablated crabs elevated the CHH mRNA level in the eyestalk, thoracic ganglion and intestinal tract tissues compared with controls. The hemolymph CHH mRNA after melatonin injection was elevated only in ablated crabs. Administration of melatonin resulted in a significant decrease in total carbohydrates and glycogen levels with an increase in phosphorylase activity levels in the hepatopancreas and muscle in intact and ablated crabs. Our findings demonstrated that melatonin can induce hyperglycemic effects in both intact and ablated crabs, suggesting that this effect is probably not mediated solely via eyestalk.

An adjoint method of data assimilation with the characteristic finite difference (CFD) scheme is applied to marine pollutant transport problems and the temporal and spatial distribution of marine pollutants are simulated. Numerical tests of two-dimensional problems of pollutant transport with two different schemes indicate that the error of CFD is smaller than that of central difference scheme (CDS). Then the inversion experiments of the initial field and the source and sink terms of pollutants are carried out. Applying CFD in the adjoint method of data assimilation cannot only reduce simulation error to get a good inversion but can also enable larger time step size to decrease computation time and improve the calculation efficiency.

Fifteen semi-synthetic derivatives of asiatic acid (AA) have been synthesized and evaluated for their biological activities. The successful modification of these compounds at the C-2, C-3, C-23 and C-28 positions was confirmed using NMR, MS and IR spectra. Further, their anti-tumor effects were evaluated in vitro using different cancer cell lines (HeLa, HepG2, B16F10, SGC7901, A549, MCF7 and PC3), while their anti-angiogenic activities were evaluated in vivo using a larval zebrafish model. Among the derivatives, compounds 4–10 showed more potent cytotoxic and anti-angiogenic effects than AA, while compounds 11–17 had significantly less effects. The new derivative 10 was also included in finished formulations to evaluate its stability using HPLC due to its potential topical use. The derivative 10 had markedly better anti-tumor activities than both AA and other derivatives, with similar stability as its parent compound AA.

We propose combining the adjoint assimilation method with characteristic finite difference scheme (CFD) to solve the aerosol transport problems, which can predict the distribution of atmospheric aerosols efficiently by using large time steps. Firstly, the characteristic finite difference scheme (CFD) is tested to compute the Gaussian hump using large time step sizes and is compared with the first-order upwind scheme (US1) using small time steps; the US1 method gets E2 error of 0.2887 using Δt=1/450, while CFD method gets a much smaller E2 of 0.2280 using a much larger time step Δt=1/45. Then, the initial distribution of PM2.5 concentration is inverted by the adjoint assimilation method with CFD and US1. The adjoint assimilation method with CFD gets better accuracy than adjoint assimilation method with US1 while adjoint assimilation method with CFD costs much less computational time. Further, a real case of PM2.5 concentration distribution in China during the APEC 2014 is simulated by using adjoint assimilation method with CFD. The simulation results are in good agreement with the observed values. The adjoint assimilation method with CFD can solve large scale aerosol transport problem efficiently.

As the major part of big data research, the audio frequency's value has not been discovered. The features of big data, and the automatic speech recognition（ASR）technology were studied, and the value was mined through the method that combined with the two technologies. Finally, the application development technical architecture and application process of audio frequency which combined with big data were given.

BackgroundUnderstanding host-pathogen interaction mechanisms helps to elucidate the entire infection process and focus on important events, and it is a promising approach for improvement of disease control and selection of treatment strategy. Time-course host-pathogen transcriptome analyses and network inference have been applied to unravel the direct or indirect relationships of gene expression alterations. However, time series analyses can suffer from absent time points due to technical problems such as RNA degradation, which limits the application of algorithms that require strict sequential sampling. Here, we introduce an efficient method using independence test to infer an independent network that is exclusively concerned with the frequency of gene expression changes.ResultsHighly resistant NL895 poplar leaves and weakly resistant NL214 leaves were infected with highly active and weakly active Marssonina brunnea, respectively, and were harvested at different time points. The independent network inference illustrated the top 1,000 vital fungus-poplar relationships, which contained 768 fungal genes and 54 poplar genes. These genes could be classified into three categories: a fungal gene surrounded by many poplar genes; a poplar gene connected to many fungal genes; and other genes (possessing low degrees of connectivity). Notably, the fungal gene M6_08342 (a metalloprotease) was connected to 10 poplar genes, particularly including two disease-resistance genes. These core genes, which are surrounded by other genes, may be of particular importance in complicated infection processes and worthy of further investigation.ConclusionsWe provide a clear framework of the interaction network and identify a number of candidate key effectors in this process, which might assist in functional tests, resistant clone selection, and disease control in the future.

To date, few studies are conducted to quantify the effects of reduced ammonium (NH4+) and oxidized nitrate (NO3-) on soil CH4 uptake and N2O emission in the subtropical forests. In this study, NH4Cl and NaNO3 fertilizers were applied at three rates: 0, 40 and 120 kg N ha(-1) yr(-1). Soil CH4 and N2O fluxes were determined twice a week using the static chamber technique and gas chromatography. Soil temperature and moisture were simultaneously measured. Soil dissolved N concentration in 0-20 cm depth was measured weekly to examine the regulation to soil CH4 and N2O fluxes. Our results showed that one year of N addition did not affect soil temperature, soil moisture, soil total dissolved N (TDN) and NH4+-N concentrations, but high levels of applied NH4Cl and NaNO3 fertilizers significantly increased soil NO3(-)-N concentration by 124% and 157%, respectively. Nitrogen addition tended to inhibit soil CH4 uptake, but significantly promoted soil N2O emission by 403% to 762%. Furthermore, NH4+-N fertilizer application had a stronger inhibition to soil CH4 uptake and a stronger promotion to soil N2O emission than NO3(-)-N application. Also, both soil CH4 and N2O fluxes were driven by soil temperature and moisture, but soil inorganic N availability was a key integrator of soil CH4 uptake and N2O emission. These results suggest that the subtropical plantation soil sensitively responses to atmospheric N deposition, and inorganic N rather than organic N is the regulator to soil CH4 uptake and N2O emission.