miR-10a-5p Regulates the Proliferation and Differentiation of Porcine Preadipocytes Targeting the KLF11 Gene.

In the swine industry, meat quality, color, and texture are influenced by the excessive differentiation of fat cells. miRNAs have emerged as integral regulators of adipose development. This study delves into the influence of miR-10a-5b on the proliferation and differentiation of pig preadipocytes. Our findings reveal that miR-10a-5b is prevalent across various tissues. It hinders preadipocyte proliferation, amplifies the expression of adipogenic genes, promotes lipid accumulation, and, as a result, advances preadipocyte differentiation. We predict that KLF11 is the target gene of miRNA. A dual-fluorescence reporter assay was conducted to validate the binding sites of miR-10a-5b on the 3'UTR of the KLF11 mRNA. Results showed that miR-10a-5b targeted KLF11 3'UTR and reduced the fluorescence activity of the dual-fluorescent reporter vector. Our research also indicates that miR-10a-5b targets and downregulates the expression of both mRNA and the protein levels of KLF11. During the differentiation of the preadipocytes, KLF11 inhibited adipose differentiation and was able to suppress the promotion of adipose differentiation by miR-10a-5b. This underscores miR-10a-5b's potential as a significant regulator of preadipocyte behavior by modulating KLF11 expression, offering insights into the role of functional miRNAs in fat deposition.

Transcriptome Analysis Revealed Potential Genes of Skeletal Muscle Thermogenesis in Mashen Pigs and Large White Pigs under Cold Stress.

Pigs are susceptible to cold stress due to the absence of brown fat caused by the partial deletion of uncoupling protein 1 during their evolution. Some local pig breeds in China exhibit potential cold adaptability, but research has primarily focused on fat and intestinal tissues. Skeletal muscle plays a key role in adaptive thermogenesis in mammals, yet the molecular mechanism of cold adaptation in porcine skeletal muscle remains poorly understood. This study investigated the cold adaptability of two pig breeds, Mashen pigs (MS) and Large White pigs (LW), in a four-day cold (4 °C) or normal temperature (25 °C) environment. We recorded phenotypic changes and collected blood and longissimus dorsi muscle for transcriptome sequencing. Finally, the PRSS8 gene was randomly selected for functional exploration in porcine skeletal muscle satellite cells. A decrease in body temperature and body weight in both LW and MS pigs under cold stress, accompanied by increased shivering frequency and respiratory frequency, were observed. However, the MS pigs demonstrated stable physiological homeostasis, indicating a certain level of cold adaptability. The LW pigs primarily responded to cold stress by regulating their heat production and glycolipid energy metabolism. The MS pigs exhibited a distinct response to cold stress, involving the regulation of heat production, energy metabolism pathways, and robust mitochondrial activity, as well as a stronger immune response. Furthermore, the functional exploration of PRSS8 in porcine skeletal muscle satellite cells revealed that it affected cellular energy metabolism and thermogenesis by regulating ERK phosphorylation. These findings shed light on the diverse transcriptional responses of skeletal muscle in LW and MS pigs under cold stress, offering valuable insights into the molecular mechanisms underlying cold adaptation in pigs.

[Effects of Microbial Fertilizer on Physicochemical Properties and Bacterial Communities of Saline Soil Under Brackish Water Irrigation].

The improvement of saline soil with microbial fertilizer has numerous advantages including high efficiency, green environmental protection, etc. At the same time, applying microbial fertilizer is an effective way to safely use brackish water. Based on the moderately saline soil in the Hetao irrigation area, four treatments of F1 (4500 kg·km-2), F2 (7500 kg·km-2), F3 (10500 kg·km-2), and CK without microbial fertilizer were applied under brackish water irrigation using Lycium barbarum as the indicator plants. The aim was to study the effects of different microbial fertilizer application rates on soil ions, soil moisture content, pH value, nutrients, and bacterial community in four key growth stages of L. barbarum (flowering stage, fruit expansion stage, full fruit stage, and deciduous stage). The results showed that, compared with that in CK, F1 only significantly decreased Na+ content in the first two growth stages (P<0.05), whereas F2 and F3 significantly decreased Na+ content in the whole growth period (P<0.05), with an average reduction of 33.66% and 57.98%, respectively, and F3 significantly increased soil moisture content (MC), organic matter (OM), alkaline hydrolysis nitrogen (AN), and available phosphorus (AP) contents (P<0.05) during the whole growth period. In the flourishing period of L. barbarum, the Shannon index of F3 increased by 4.41% compared with that of CK. The dominant bacterial phyla in the soil were Proteobacteria, Bacteroidetes, and Actinobacteria, and the dominant bacterial genera were Sphingomonas and Pseudomonas. The most abundant functions of bacterial communities in the study area were chemoheterotrophy and aerobic chemoheterotrophy, with an average relative abundance of 15.07% and 13.16%, respectively. The application of microbial fertilizer increased the chitinolysis function and chloroplast functions of soil bacteria, which F2 increased to the highest degree. Canonical correlation analysis (CCA) showed that MC, Na+, and OM were important factors affecting the composition of the bacterial community. The correlation heat map showed that MC was positively correlated with Planctomycetes (P<0.01), and Gp6 was positively correlated with AN (P<0.01). Compared with that in CK, the F3 treatment increased the relative abundance of Gp6 and optimized the community structure during the growth period. In conclusion, the application of 10500 kg·km-2 microbial fertilizer (F3 treatment) under brackish water irrigation could significantly reduce soil salinity, increase nutrients, and improve the diversity of the soil bacterial community structure, which is conducive to the safe utilization of brackish water and the maintenance of soil ecological health.

Corrigendum: NanoSIMS analysis of water content in bridgmanite at the micron scale: an experimental approach to probe water in Earth's deep mantle.

[This corrects the article DOI: 10.3389/fchem.2023.1166593.].

NanoSIMS analysis of water content in bridgmanite at the micron scale: An experimental approach to probe water in Earth's deep mantle.

Water, in trace amounts, can greatly alter chemical and physical properties of mantle minerals and exert primary control on Earth's dynamics. Quantifying how water is retained and distributed in Earth's deep interior is essential to our understanding of Earth's origin and evolution. While directly sampling Earth's deep interior remains challenging, the experimental technique using laser-heated diamond anvil cell (LH-DAC) is likely the only method available to synthesize and recover analog specimens throughout Earth's lower mantle conditions. The recovered samples, however, are typically of micron sizes and require high spatial resolution to analyze their water abundance. Here we use nano-scale secondary ion mass spectrometry (NanoSIMS) to characterize water content in bridgmanite, the most abundant mineral in Earth's lower mantle. We have established two working standards of natural orthopyroxene that are likely suitable for calibrating water concentration in bridgmanite, i.e., A119(H2O) = 99 ± 13 µg/g (1SD) and A158(H2O) = 293 ± 23 µg/g (1SD). We find that matrix effect among orthopyroxene, olivine, and glass is less than 10%, while that between orthopyroxene and clinopyroxene can be up to 20%. Using our calibration, a bridgmanite synthesized by LH-DAC at 33 ± 1 GPa and 3,690 ± 120 K is measured to contain 1,099 ± 14 µg/g water, with partition coefficient of water between bridgmanite and silicate melt ∼0.025, providing the first measurement at such condition. Applying the unique analytical capability of NanoSIMS to minute samples recovered from LH-DAC opens a new window to probe water and other volatiles in Earth's deep mantle.

[Effects of Phosphogypsum and Suaeda salsa on the Soil Moisture, Salt, and Bacterial Community Structure of Salinized Soil].

The improvement of saline soil is an important issue that cannot be ignored in the farmland soil environment. The change in soil salinity will inevitably affect the soil bacterial community. This experiment was based on moderately saline soil in the Hetao Irrigation Area, conducted by applying phosphogypsum (LSG), interplanting Suaeda salsa with Lycium barbarum (JP) and applying phosphogypsum and interplanting S. salsa with L. barbarum (LSG+JP),and the local unimproved soil of a L. barbarum orchard was used as the control (CK), to explore the effects of different improvement methods on soil moisture, salinity, nutrients, and bacterial community structure diversity during the growth period of L. barbarum. The results showed that compared with that under CK, the LSG+JP treatment significantly decreased the soil EC value and pH value from the flowering stage to the deciduous stage (P<0.05), with an average decrease of 39.96% and 7.25%, respectively; the LSG+JP treatment significantly increased soil organic matter (OM) and available phosphorus (AP) content during the whole growth period (P<0.05), with an average annual increase of 81.85% and 203.50%, respectively. The total nitrogen (TN) content was significantly increased in the flowering and deciduous stages (P<0.05), with an annual average increase of 48.91%. The Shannon index of LSG+JP in the early stage of improvement was increased by 3.31% and 6.54% compared with that of CK, and the Chao1 index was increased by 24.95% and 43.26% compared with that of CK, respectively. The dominant bacteria in the soil were Proteobacteria, Bacteroidetes, Actinobacteria, and Acidobacteria, and the dominant genus was Sphingomonas. Compared with that in CK, the relative abundance of Proteobacteria in the improved treatment increased by 0.50%-16.27% from the flowering stage to the deciduous stage, and the relative abundance of Actinobacteria in the improved treatment increased by 1.91%-4.98% compared with that in CK in the flowering and full-fruit stages. Redundancy analysis (RDA) results showed that pH, water content (WT), and AP were important factors affecting bacterial community composition, and the correlation heatmap showed that Proteobacteria, Bacteroidetes, and EC values were significantly negatively correlated (P<0.001); Actinobacteria and Nitrospirillum were significantly negatively correlated with EC values (P<0.01). In conclusion, the application of phosphogypsum and interplanting S. salsa with L. barbarum (LSG+JP) could significantly reduce soil salinity, increase nutrients, and improve the diversity of soil bacterial community structure, which is beneficial to the long-term improvement of saline soil in the Hetao Irrigation Area and the maintenance of soil ecological health.

The formulation of irrigation and nitrogen application strategies under multi-dimensional soil fertility targets based on preference neural network.

With the aim of improving soil fertility, it is of great significance to put forward optimal irrigation and nitrogen fertilizer application strategies for improving land productivity and alleviating non-point source pollution effects. To overcome this task, a 6-hidden layer neural network with a preference mechanism, namely Preference Neural network (PNN), has been developed in this study based on the field data from 2018 to 2020. PNN takes soil total nitrogen, organic matter, total salt, pH, irrigation time and target soil depth as input, and irrigation amount and nitrogen application rate (N rate) as output, and the prior preference matrix was used to adjust the learning of weight matrix of each layer. The outcomes indicated that the predictive accuracy of PNN for irrigation amount were (R2 = 0.913, MAE = 0.018, RMSE = 0.022), and for N rate were (R2 = 0.943, MAE = 0.009, RMSE = 0.011). The R2 predicted by PNN at the irrigation amount and N rate were 40.03% to more than 99% and 40.33% to more than 99% higher than those obtained using support vector regression (SVR), linear regression (LR), logistic regression (LOR) and traditional back propagation neural network (BPNN), respectively. In addition, compared with the neural network (Reverse Multilayer Perceptron, RMLP) with the same structure but no preference structure, the R2 of the predicted irrigation amount and N rate by PNN increased by 25.81% and 27.99%, respectively. The results showed that, through the irrigation of 93 to 102, 92 to 98 and 92 to 98 mm, along with nitrogen applications of 65 to 71, 64 to 73 and 72 to 81 kg/hm2 at 17, 59 and 87 days after sowing, respectively, the organic matter, total nitrogen, total salt content and pH of the soil would reach high fertility levels simultaneously.

Effects of Low-Ambient-Temperature Stimulation on Modifying the Intestinal Structure and Function of Different Pig Breeds.

Ambient temperature (Ta) fluctuation is a key factor affecting the growth performance and economic returns of pigs. However, whether the response of intestinal structure and function are related to pig breeds in low Ta has not been investigated yet. In this study, Large White (LW) pigs, Jinfen White (JFW) pigs and Mashen (MS) pigs were raised in artificial climate chambers under normal Ta (25 °C) and low Ta (4 °C) for 96 h. Afterwards, the decrease in body temperature and complete blood counts (CBC) of all pigs were measured. Hematoxylin-eosin, immunohistochemical staining, qPCR and ELISA were used to investigate their intestinal mucosa integrity and inflammatory response. The results showed that MS pigs could maintain a normal body temperature and villus structure after 4 °C stimulation compared with those of LW and JFW pigs. Villus height and villus height/crypt depth of MS pigs were significantly higher than those of LW and JFW pigs at 4 °C. Low-Ta stimulation increased the digestion of carbohydrates of all pigs. Meanwhile, low Ta enhanced the activity of lipase in LW pigs and increased trypsin activity in MS and JFW pigs. Furthermore, low-Ta stimulation significantly downregulated the protein of tight junction and upregulated the mRNA expression of inflammatory cytokines in MS pigs. MS pigs also showed stronger spleen immune function at 4 °C. These results indicated that the local MS pig breed had stronger intestinal function in low Ta by producing a stronger inflammatory response, which lays the foundation for further study on the mechanism of cold tolerance in pigs.

Effects of straw mulching combined with nitrogen application on soil organic matter content and atrazine digestion.

To explore the response of atrazine (AT) degradation rate, soil organic matter (SOM) distribution and the relationship between them to straw mulching and nitrogen application, field experiments were conducted to study the distribution of SOM content and AT degradation rate under different straw returning modes combined with nitrogen fertilization in 2 years in Hetao Irrigation District. No (N0), low (NL), medium (NM), and high (NH) levels of N fertilization were incorporated into the soil combined with the surface coverage straw (Treatment BN0, BNL, BNM, BNH, respectively) and the deeply buried straw (Treatment SN0, SNL, SNM, SNH, respectively). The traditional cultivation was used as a control treatment (Treatment CK). The results showed that SOM content of Treatment B was accumulated in 0~20 cm soil layer. The largest SOM content of Treatments B in 0~20 cm soil layer was found in BNH treatment, with an average increase of 14.2% and 24.1% significantly when compared with those in CK and SNH (P < 0.05), respectively. The SOM content of Treatments B increased with the increase of nitrogen application and decreased with the deepening of soil depth. The SOM content of Treatment S was accumulated in the soil layer (20~40 cm and 40~60 cm) near the inter-layer. With the increase of nitrogen application and depth of soil layer, the SOM content increased firstly and then decreased. SNM had the largest SOM content in the soil layer of 20~40 cm and 40~60 cm, with an average increase of 82.6% and 67.7% when compared with Treatment CK (P < 0.05). In the soil layer over 60 cm, there was no significant difference in SOM content of different straw returning methods under the same nitrogen level (P > 0.05). Straw returning methods and nitrogen application level significantly affected AT digestion rate and digestion half-life, with significant differences among treatments (P < 0.05). It found that treatment SNM had the highest digestion rate and the shortest half-life of AT. Compared with Treatment CK, the digestion rate of Treatment SNM was increased by 5.3% on average, and the half-life was shortened by 3.9 days on average. Single regression and stepwise regression analysis of the half-life of AT degradation and SOM content in different soil layers (0~20 cm and 20~40 cm) showed that the degradation of AT was greatly affected by SOM content of 20~40 cm soil layer. Based on the comprehensive analysis, the effect of straw deep burial combined with medium nitrogen application rate (Treatment SNM) was best, which could achieve the goal of increasing SOM content and shortening the half-life of AT digestion. The research provided a technical support for straw resource utilization, alleviated AT pollution and improved farmland ecological environment in Hetao Irrigated District.

The clonal expression genes associated with poor prognosis of liver cancer.

The extensive spatial genomic intratumor heterogeneity (ITH) in liver cancer hindered treatment development and limited biomarker design. Early events that drive tumor malignant transformation in tumor founder cells are clonally present in all tumor cell populations, which provide stable biomarkers for the localization of tumor cells and patients' prognosis. In the present study, we identified the recurrently clonal somatic mutations and copy number alterations (CNAs) (893 clonal somatic mutations and 6,617 clonal CNAs) in 353 liver cancer patients from The Cancer Genome Atlas (TCGA) and evaluated their prognosis potential. We showed that prognosis-related clonal alterations might play essential roles in tumor evolution. We identified 32 prognosis related clonal alterations differentially expressed between paired normal and tumor samples, that their expression was cross-validated by three independent cohorts (50 paired samples in TCGA, 149 paired samples in GSE76297, and 9 paired samples in SUB6779164). These clonal expression alterations were also significantly correlated with clinical phenotypes. Using stepwise regression, we identified five (UCK2, EFNA4, KPAN2, UBE2T, and KIF14) and six (MCM10, UCK2, IQGAP3, EFNA4, UBE2T, and KPNA2) clonal expression alterations for recurrence and survival model construction, respectively. Furthermore, in 10 random repetitions, we showed strong applicability of the multivariate Cox regression models constructed based on the clonal expression genes, which significantly predicted the outcomes of the patients in all the training and validation sets. Taken together, our work may provide a new avenue to overcome spatial ITH and refine biomarker design across cancer types.

MiR-23b Promotes Porcine Preadipocyte Differentiation via SESN3 and ACSL4.

Genetic improvement of pork quality is one of the hot topics in pig germplasm innovation. Backfat thickness and intramuscular fat content are important indexes of meat quality. MiRNAs are becoming recognized as a crucial regulator of adipose development. Therefore, it is crucial to understand how miR-23b regulates fat metabolism at the molecular level. In the present study, Oil Red O staining, and Western blot were used to evaluate the effect of miR-23b on the differentiation of porcine preadipocytes. Dual-luciferase reporter gene assay, pulldown, and RIP were used to reveal the mechanism of miR-23b regulating cell differentiation. The findings demonstrated that miR-23b promotes the expression of adipogenic factors and increases the content of lipid droplets, thus promoting the differentiation of preadipocytes. Further research found that miR-23b can directly bind to the 3'UTR of SESN3 to regulate adipogenic differentiation. In addition, it was speculated that miR-23b controls cell differentiation by positively regulating the expression of ACSL4 in other ways. Here, we demonstrate that miR-23b promotes the differentiation of porcine preadipocytes by targeting SESN3 and promoting the expression of ACSL4. The present study is meaningful to the improvement of pork quality and the development of animal husbandry.

[Temporal and Spatial Variation Characteristics and Driving Factors of Nitrogen of Shallow Groundwater in Hetao Irrigation District].

As an important food production area in the north of China, the Hetao plain is extremely vulnerable to nitrate pollution caused by agricultural production activities and additional factors. Thus, it is of great significance for the environmental protection and rational use of groundwater to detect the current situation of groundwater nitrate pollution, temporal and spatial evolution characteristics, and main influencing factors in the Hetao irrigation district. We selected the Wualte irrigation area as the study area, and the characteristics of temporal and spatial changes in groundwater nitrate concentration and the main influencing factors in this area were explored. We used statistical analysis to analyze the nitrogen content of groundwater in the study area, and the Piper three-line diagram was used to explore the characteristics of chemical composition and evolution; furthermore, we used ion ratio and correlation analysis methods to explore the source of NO3--N in groundwater. The results showed that NO3--N was the main existing form of nitrogen in the Wulate irrigation area, and its concentration varied from 0.01 to 60.00 mg·L-1, with an exceeding standard rate of 10.50%. In terms of time, the characteristic of time change was that the NO3--N concentration in August of groundwater was the highest (average 6.61 mg·L-1), followed by that in October (6.22 mg·L-1) and November (6.25 mg·L-1), and that in March (average value of 1.77 mg·L-1) was the lowest. With the influence of rainfall and irrigation, the NO3--N in the soil was infiltrated into the groundwater, showing the characteristic that wet season and concentrated irrigation periods were higher than those in other periods. Spatially, it appeared as southwest (8.87 mg·L-1)>northwest (4.25 mg·L-1)>east (0.89 mg·L-1), mainly due to the original geological conditions, land use, and domestic waste stacking. In addition, the concentration of NO3--N of groundwater in the study area was closely related to the depth of groundwater and redox conditions but was relatively less affected by the concentration of water chemical ions. Therefore, identifying the temporal and spatial distribution characteristics and main sources of groundwater nitrogen pollution can provide a scientific basis for scientific fertilization, groundwater nitrate pollution control, and water safety.

Glutamic-oxaloacetic transaminase 1 regulates adipocyte differentiation by altering nicotinamide adenine dinucleotide phosphate content.

OBJECTIVE: This study was performed to examine whether the porcine glutamic-oxaloacetic transaminase 1 (GOT1) gene has important functions in regulating adipocyte differentiation. METHODS: Porcine GOT1 knockout and overexpression vectors were constructed and transfected into the mouse adipogenic 3T3-L1 cells. Lipid droplets levels were measured after 8 days of differentiation. The mechanisms through which GOT1 participated in lipid deposition were examined by measuring the expression of malate dehydrogenase 1 (MDH1) and malic enzyme (ME1) and the cellular nicotinamide adenine dinucleotide phosphate (NADPH) content. RESULTS: GOT1 knockout significantly decreased lipid deposition in the 3T3-L1 cells (p<0.01), whereas GOT1 overexpression significantly increased lipid accumulation (p<0.01). At the same time, GOT1 knockout significantly decreased the NADPH content and the expression of MDH1 and ME1 in the 3T3-L1 cells. Overexpression of GOT1 significantly increased the NADPH content and the expression of MDH1 and ME1, suggesting that GOT1 regulated adipocyte differentiation by altering the NADPH content. CONCLUSION: The results preliminarily revealed the effector mechanisms of GOT1 in regulating adipose differentiation. Thus, a theoretical basis is provided for improving the quality of pork and studies on diseases associated with lipid metabolism.

FNDC5 Promotes Adipogenic Differentiation of Primary Preadipocytes in Mashen Pigs.

Fibronectin type III domain-containing protein 5 (FNDC5) plays an important role in fat deposition, which can be cut to form Irisin to promote fat thermogenesis, resulting in a decrease in fat content. However, the mechanism of FNDC5 related to fat deposition in pigs is still unclear. In this research, we studied the expression of FNDC5 on different adiposes and its function in the adipogenic differentiation of primary preadipocytes in Mashen pigs. The expression pattern of FNDC5 was detected by qRT-PCR and Western blotting in Mashen pigs. FNDC5 overexpression and interference vectors were constructed and transfected into porcine primary preadipocytes by lentivirus. Then, the expression of key adipogenic genes was detected by qRT-PCR and the content of lipid droplets was detected by Oil Red O staining. The results showed that the expression of FNDC5 in abdominal fat was higher than that in back subcutaneous fat in Mashen pigs, whereas the expression in back subcutaneous fat of Mashen pigs was significantly higher than that of Large White pigs. In vitro, FNDC5 promoted the adipogenic differentiation of primary preadipocytes of Mashen pigs and upregulated the expression of genes related to adipogenesis, but did not activate the extracellular signal-regulated kinase (ERK) signaling pathway. This study can provide a theoretical basis for FNDC5 in adipogenic differentiation in pigs.

Comprehensive analysis of differentially expressed circRNAs and ceRNA regulatory network in porcine skeletal muscle.

BACKGROUND: Circular RNA (circRNA), a novel class of non-coding RNA, has a closed-loop structure with important functions in skeletal muscle growth. The purpose of this study was to investigate the role of differentially expressed circRNAs (DEcircRNAs), as well as the DEcircRNA-miRNA-mRNA regulatory network, at different stages of porcine skeletal muscle development. Here, we present a panoramic view of circRNA expression in porcine skeletal muscle from Large White and Mashen pigs at 1, 90, and 180 days of age. RESULTS: We identified a total of 5819 circRNAs. DEcircRNA analysis at different stages showed 327 DEcircRNAs present in both breeds. DEcircRNA host genes were concentrated predominately in TGF-ß, MAPK, FoxO, and other signaling pathways related to skeletal muscle growth and fat deposition. Further prediction showed that 128 DEcircRNAs could bind to 253 miRNAs, while miRNAs could target 945 mRNAs. The constructed ceRNA network plays a vital role in skeletal muscle growth and development, and fat deposition. Circ_0015885/miR-23b/SESN3 in the ceRNA network attracted our attention. miR-23b and SESN3 were found to participate in skeletal muscle growth regulation, also playing an important role in fat deposition. Using convergent and divergent primer amplification, RNase R digestion, and qRT-PCR, circ_0015885, an exonic circRNA derived from Homer Scaffold Protein 1 (HOMER1), was confirmed to be differentially expressed during skeletal muscle growth. In summary, circ_0015885 may further regulate SESN3 expression by interacting with miR-23b to function in skeletal muscle. CONCLUSIONS: This study not only enriched the circRNA library in pigs, but also laid a solid foundation for the screening of key circRNAs during skeletal muscle growth and intramural fat deposition. In addition, circ_0015885/miR-23b/SESN3, a new network regulating skeletal muscle growth and fat deposition, was identified as important for increasing the growth rate of pigs and improving meat quality.

Integrative analysis of the gastric cancer long non-coding RNA-associated competing endogenous RNA network.

Gastric cancer (GC) is a common type of cancer, and identification of novel diagnostic biomarkers associated with this disease is important. The present study aimed to identify novel diagnostic biomarkers associated with the prognosis of GC, using an integrated bioinformatics approach. Differentially expressed long non-coding RNAs (lncRNAs) associated with GC were identified using Gene Expression Omnibus datasets (GSE58828, GSE72305 and GSE99416) and The Cancer Genome Atlas database. A competing endogenous RNA network that incorporated five lncRNAs [long intergenic non-protein coding RNA 501 (LINC00501), LINC00365, SOX21 antisense divergent transcript 1 (SOX21-AS1), GK intronic transcript 1 (GK-IT1) and DLEU7 antisense RNA 1 (DLEU7-AS1)], 29 microRNAs and 114 mRNAs was constructed. Gene Ontology and protein-protein interaction network analyses revealed that these lncRNAs may be involved in 'biological regulation', 'metabolic process', 'cell communication', 'developmental process', 'cell proliferation', 'reproduction' and the 'cell cycle'. The results of receiver operating characteristic curve analysis demonstrated that LINC00501 (AUC=0.819), LINC00365 (AUC=0.580), SOX21-AS1 (AUC=0.736), GK-IT1 (AUC=0.823) and DLEU7-AS1 (AUC=0.932) had the potential to become valuable diagnostic biomarkers for GC. Associations with clinicopathological characteristics demonstrated that LINC00501 expression was significantly associated with sex (P=0.015) and tumor grade (P=0.022). Furthermore, LINC00365 expression was significantly associated with lymph node metastasis (P=0.025). Gene set enrichment analysis revealed that LINC00501, LINC00365 and SOX21-AS1 were enriched in signaling pathways associated with GC. Reverse transcription-quantitative PCR analysis demonstrated that LINC00501 expression (P=0.043) was significantly upregulated in GC tissues, whereas the expression levels of LINC00365 (P=0.033) and SOX21-AS1 (P=0.037) were significantly downregulated in GC tissues. Taken together, the results of the present study suggest that LINC00501, LINC00365, SOX21-AS1, GK-IT1 and DLEU7-AS1 may be used as novel diagnostic biomarkers for GC, and may be functionally associated with GC development and progression.

[Effects of Straw Mulching and Nitrogen Reduction on the Distribution of Soil Nitrogen and Groundwater Nitrogen Pollution].

To explore the effects of straw mulching and reduced nitrogen fertilization on the temporal and spatial patterns of soil nitrogen, groundwater nitrogen pollution, and summer maize yield, field experiments were carried out in the Hetao irrigation district in 2017 and 2018. The experiment involved the following seven treatments:a control (CK) treatment involving conventional fertilization and traditional tillage, and conventional nitrogen applications reduced by 30% (N1), 20% (N2), and 10% (N3) coupled with either straw surface covering (B) or deep straw burial (S). The results showed that the distribution of soil nitrogen in the CK treatment varied depending on soil depth, with an overall decreasing trend. In the 0-20 cm soil layer under straw surface covering (B) treatments, soil nitrogen was superficially accumulated. NO3--N and NH4+-N content increased by an average of 22.2% and 42.7% compared to the CK treatment, respectively, which decreased significantly at first and then increased slightly with depth. In the 20-40 cm deep soil layer under the deep straw burial (S) treatments, soil nitrogen accumulated and the content of NO3--N and NH4+-N increased by an average of 29.8% and 48.1%, respectively, compared to the CK treatment. Nitrogen accumulation first and then decreased significantly with depth. Nitrogen accumulation under the different straw mulching regimes increased with an increase in the application of reduced nitrogen. After the harvest of summer maize, the accumulation of NO3--N and NH4+-N in the >80 cm soil layer under the B treatments was 19.9%-58.2% and 31.1%-61.7% lower than that of the CK treatment, respectively. This compared to reductions of 36.7%-70.9% and 82.6%-89.2% for the S treatments, respectively. Only the BN3 treatment increased accumulation compared with CK by 0.4% on average, while the SN2 treatment resulted in a 9.3% increase. Summer maize yield and relative indexes were also improved relative to the other treatments. Nonlinear fitting of yield and application reduction showed that deep straw burial was better than surface covering at increasing summer maize production. The effect of deep straw burial and 14%-20% application reduction was better. Straw mulching with reduced nitrogen fertilization can limit nitrogen leaching and thereby reduce the risk of groundwater pollution. After the harvest, groundwater quality was classified in the Ⅱ class, with the risk of nitrogen contamination being lowest under deep straw burial with>20% reduced nitrogen fertilization. These observations show that deep straw deep alongside 14%-20% application reduction could effectively alleviate nitrogen leaching and reduce the risk of nitrogen pollution in groundwater. This approach can help improve the ecological environment and summer maize yields in the Hetao irrigation district.

The effects of straw mulching combined with nitrogen applications on the root distributions and nitrogen utilization efficiency of summer maize.

To provide an appropriate tillage fertilization model for improving N utilization efficiency and increasing production, the field experiments were conducted to study the effects on root distributions and N utilization efficiency of summer maize involving different straw mulching modes combined with N fertilization. No (N0), low (N1), medium (N2), and high (N3) levels of N fertilization were incorporated into soil combined with the surface coverage straw (Treatment B) and the deeply buried straw (Treatment S). The traditional cultivation was used as control treatment. The results shown that treatments S had significantly promoted deep root growth, and the root length density (RLD) increased with increases in N application rate. SN2 and SN3 treatments' average RLD were significantly increased by 67.5% and 68.1% in the greater than 40 cm soil layers. While the Treatment B had significantly increased the RLD in 0 -30 cm soil layers only. With increases in N application rate, the effect on summer maize yields increase under Treatment B were not significantly, and only BN3 increased by 0.4%, while under Treatments S were found to first increase, and then decrease. The apparent recovery efficiency of applied N, N uptake and summer maize yield of SN2 had increased by 66.8%, 20.4%, and 9.3%. Therefore the rational tillage fertilization model was deeply buried straw combined with medium N fertilizer in Hetao Irrigation District.

Reconstruction of skull and dural defects using anterolateral thigh flaps: A STROBE-compliant article.

It is difficult to repair large skull and dural defects. We observed the therapeutic effects of anterolateral thigh flaps with vascular fascia lata for repairing large skull and dural defects.From December 2008 to June 2019, we repaired large skull and dural defects for 28 cases including 12 cases with scalp malignant tumor and 16 cases requiring removal of titanium mesh which had been once placed due to craniocerebral trauma. The scalp malignant tumor invaded full-thickness skull in 12 cases; and invaded cervical lymph nodes, dura mater or brain tissue in 3 cases. In the 12 cases with scalp malignant tumor, the scalp defects of 12 cm × 9 cm to 22 cm × 18 cm and skull defects of 9 cm × 7 cm to 15 cm × 12 cm after radical tumor resection were repaired using anterolateral thigh flaps of 14 cm × 11 cm to 23 cm × 19 cm with fascia lata of 10 cm × 8 cm to 16 cm × 12 cm. Postoperative radiotherapy and chemotherapy were also performed in the 3 cases with tumor metastasis. In the 16 cases requiring removal of titanium mesh, the skull and dural defects of 8 cm × 7 cm to 15 cm × 11 cm after removal of titanium mesh were repaired using anterolateral thigh flaps of 10 cm × 8 cm to 16 cm × 12 cm.In all cases, the transplanted anterolateral thigh flap with fascia lata survived after surgery and no vascular crisis occurred. During the followup of 8 months to 9 years, the flap appearance in the head-repaired area was fine, no external hernia of brain tissue occurred, the appearance of the femoral donor site was acceptable, and femoral muscle strength and movements were normal in all cases. The 12 cases with scalp malignant tumor had no local recurrence or distant metastasis.Repairing the skull and dural defects caused by radical surgery for scalp malignant tumor or removal of titanium mesh using anterolateral thigh flaps with vascular fascia lata, is effective. The appearance in the head-repaired area is fine without external hernia of brain tissue.

A Novel Change Detection Method for Natural Disaster Detection and Segmentation from Video Sequence.

Change detection (CD) is critical for natural disaster detection, monitoring and evaluation. Video satellites, new types of satellites being launched recently, are able to record the motion change during natural disasters. This raises a new problem for traditional CD methods, as they can only detect areas with highly changed radiometric and geometric information. Optical flow-based methods are able to detect the pixel-based motion tracking at fast speed; however, they are difficult to determine an optimal threshold for separating the changed from the unchanged part for CD problems. To overcome the above problems, this paper proposed a novel automatic change detection framework: OFATS (optical flow-based adaptive thresholding segmentation). Combining the characteristics of optical flow data, a new objective function based on the ratio of maximum between-class variance and minimum within-class variance has been constructed and two key steps are motion detection based on optical flow estimation using deep learning (DL) method and changed area segmentation based on an adaptive threshold selection. Experiments are carried out using two groups of video sequences, which demonstrated that the proposed method is able to achieve high accuracy with F1 value of 0.98 and 0.94, respectively.