Aberrant Splicing of COL4A5 Intronic Variant Contribute to the Pathogenesis of X-Linked Alport Syndrome: A Case Series.

Introduction: X-linked Alport syndrome (XLAS) is caused by pathogenic variants in COL4A5 which lead to abnormalities of the glomerular basement membrane (GBM) structural and is characterized by progressive kidney disease, hearing loss, and ocular abnormalities. The aim of this study was to identify gene mutations in a Chinese family with XLAS by whole-exome sequencing (WES) and verified the pathogenicity of the mutation in vitro experiments. Case Presentation: A five-generation pedigree with a total of 49 family members originating from Hainan province of China was investigated in this study. The proband was a 23-year-old male who developed microscopic hematuria, proteinuria and end-stage kidney disease (ESKD) at age 17. WES identified a novel splicing mutation c.321+5G>A of COL4A5, which cause exon skip. Further co-segregation analysis confirmed that this mutation exists in relatives who had renal abnormalities using Sanger sequencing. According to American College of Medical Genetics and Genomics guidelines (ACMG), the mutation was determined to be of uncertain significance (VUS). In vitro splicing experiments have shown that the COL4A5 variant induces aberrant mRNA splicing and transcript deletion. Conclusion: We identified a novel intronic COL4A5 pathogenic mutation (c.321+5G>A) in a Chinese XLAS family and described the phenotypes of affected relatives. This study expands the mutation spectrum of COL4A5 gene in XLAS and demonstrates the importance of gene screening for AS.

Targeted next-generation sequencing and long-read HiFi sequencing provide novel insights into clinically significant KLF1 variants.

BACKGROUND: Krüppel-like factor 1 (KLF1), a crucial erythroid transcription factor, plays a significant role in various erythroid changes and haemolytic diseases. The rare erythrocyte Lutheran inhibitor (In(Lu)) blood group phenotype serves as an effective model for identifying KLF1 hypomorphic and loss-of-function variants. In this study, we aimed to analyse the genetic background of the In(Lu) phenotype in a population-based sample group by high-throughput technologies to find potentially clinically significant KLF1 variants. RESULTS: We included 62 samples with In(Lu) phenotype, screened from over 300,000 Chinese blood donors. Among them, 36 samples were sequenced using targeted Next Generation Sequencing (NGS), whereas 19 samples were sequenced using High Fidelity (HiFi) technology. In addition, seven samples were simply sequenced using Sanger sequencing. A total of 29 hypomorphic or loss-of-function variants of KLF1 were identified, 21 of which were newly discovered. All new variants discovered by targeted NGS or HiFi sequencing were validated through Sanger sequencing, and the obtained results were found to be consistent. The KLF1 haplotypes of all new variants were further confirmed using clone sequencing or HiFi sequencing. The lack of functional KLF1 variants detected in the four samples indicates the presence of additional regulatory mechanisms. In addition, some samples exhibited BCAM polymorphisms, which encodes antigens of the Lutheran (LU) blood group system. However, no BCAM mutations which leads to the absence of LU proteins were detected. CONCLUSIONS: High-throughput sequencing methods, particularly HiFi sequencing, were introduced for the first time into genetic analysis of the In(Lu) phenotype. Targeted NGS and HiFi sequencing demonstrated the accuracy of the results, providing additional advantages such as simultaneous analysis of other blood group genes and clarification of haplotypes. Using the In(Lu) phenotype, a powerful model for identifying hypomorphic or loss-of-function KLF1 variants, numerous novel variants have been detected, which have contributed to the comprehensive understanding of KLF1. These clinically significant KLF1 mutations can serve as a valuable reference for the diagnosis of related blood cell diseases.

A Predictive Network-Based Immune Checkpoint Blockade Immunotherapeutic Signature Optimizing Patient Selection and Treatment Strategies.

Immune checkpoint blockade (ICB) therapy has brought significant advancements to the field of oncology. However, the diverse responses among patients highlight the need for more accurate predictive tools. In this study, insights are drawn from tumor-immunology pathways, and a novel network-based ICB immunotherapeutic signature, termed ICBnetIS, is constructed. The signature is derived from advanced biological network-based computational strategies involving co-expression networks and molecular interactions networks. The efficacy of ICBnetIS is established through its association with enhanced patient survival and a robust immune response characterized by diverse immune cell infiltration and active anti-tumor immune pathways. The validation process positions ICBnetIS as an effective tool in predicting responses to ICB therapy, analyzing ICB data from a broad collection of over 700 samples from multiple cancer types of more than 15 datasets. It achieves an aggregated prediction AUC of 0.784, which outperforms the other nine renowned immunotherapeutic signatures, indicating the superior predictive capability of ICBnetIS. To sum up, the findings suggest ICBnetIS as a potent tool in predicting ICB therapy responses, offering significant implications for patient selection and treatment optimization in oncology. The study highlights the role of ICBnetIS in advancing personalized treatment strategies, potentially transforming the clinical landscape of ICB therapy.

The effect of modifier and a water-soluble fertilizer on two forages grown in saline-alkaline soil.

Saline-alkali soil significantly impairs crop growth. This research employs the impacts of the modifier and water-soluble fertilizer, as well as their interaction, on the root systems of alfalfa and leymus chinensis in saline-alkali soil. The results exhibit that the hydrochar source modifier effectively enhances the root growth of both forage species. There are certain improvements in the root growth indicators of both crops at a dosage of 20 g/kg. Root enzyme activity and rhizosphere soil enzyme activity are enhanced in alfalfa, showing significant improvements in the first planting compared to the second planting. The application of water-soluble fertilizers also promotes root growth and root dehydrogenase activity. The root dehydrogenase activity of alfalfa and leymus chinensis are enhanced 62.18% and 10.15% in first planting than that of blank, respectively. Additionally, the two-factor variance analysis revealed a correlation between rhizosphere soil enzyme activity and changes in root traits. Higher rhizosphere soil enzyme activity is observed in conjunction with better root growth. The combined application of a modifier and water-soluble fertilizer has demonstrated a significant interaction effect on various aspects of the first planting of alfalfa and leymus chinensis. Moreover, the combined application of the modifier and water-soluble fertilizer has yielded superior results when compared to the individual application of either the modifier or the water-soluble fertilizer alone. This combined approach has proven effective in improving saline-alkali soil conditions and promoting crop growth in such challenging environments.

Preimplantation genetic testing in the current era, a review.

BACKGROUND: Preimplantation genetic testing (PGT), also referred to as preimplantation genetic diagnosis (PGD), is an advanced reproductive technology used during in vitro fertilization (IVF) cycles to identify genetic abnormalities in embryos prior to their implantation. PGT is used to screen embryos for chromosomal abnormalities, monogenic disorders, and structural rearrangements. DEVELOPMENT OF PGT: Over the past few decades, PGT has undergone tremendous development, resulting in three primary forms: PGT-A, PGT-M, and PGT-SR. PGT-A is utilized for screening embryos for aneuploidies, PGT-M is used to detect disorders caused by a single gene, and PGT-SR is used to detect chromosomal abnormalities caused by structural rearrangements in the genome. PURPOSE OF REVIEW: In this review, we thoroughly summarized and reviewed PGT and discussed its pros and cons down to the minutest aspects. Additionally, recent studies that highlight the advancements of PGT in the current era, including their future perspectives, were reviewed. CONCLUSIONS: This comprehensive review aims to provide new insights into the understanding of techniques used in PGT, thereby contributing to the field of reproductive genetics.

Optimization of FFPE preparation and identification of gene attributes associated with RNA degradation.

Formalin-fixed paraffin-embedded (FFPE) tissues are widely available specimens for clinical studies. However, RNA degradation in FFPE tissues often restricts their utility. In this study, we determined optimal FFPE preparation conditions, including tissue ischemia at 4°C (<48 h) or 25°C for a short time (0.5 h), 48-h fixation at 25°C and sampling from FFPE scrolls instead of sections. Notably, we observed an increase in intronic reads and a significant change in gene rank based on expression level in the FFPE as opposed to fresh-frozen (FF) samples. Additionally, we found that more reads were mapped to genes associated with chemical stimulus in FFPE samples. Furthermore, we demonstrated that more degraded genes in FFPE samples were enriched in genes with short transcripts and high free energy. Besides, we found 40 housekeeping genes exhibited stable expression in FF and FFPE samples across various tissues. Moreover, our study showed that FFPE samples yielded comparable results to FF samples in dimensionality reduction and pathway analyses between case and control samples. Our study established the optimal conditions for FFPE preparation and identified gene attributes associated with degradation, which would provide useful clues for the utility of FFPE tissues in clinical practice and research.

Space-CNN: a decision classification method based on EEG signals from different brain regions.

Decision-making plays a critical role in an individual's interpersonal interactions and cognitive processes. Due to the issue of strong subjectivity in the classification research of art design decisions, we utilize the relatively objective electroencephalogram (EEG) to explore design decision problems. However, different regions of the brain do not have the same influence on the design decision classification, so this paper proposes a spatial feature based convolutional neural network (space-CNN) to explore the problem of decision classification of EEG signals from different regions. We recruit 16 subjects to collect their EEG data while viewing four stimulation patterns. After noise reduction of the raw data by discrete wavelet transform (DWT), the EEG image is generated by combining it with the spatial features of the EEG signal, which is used as the input of CNN. Our experimental results show that the degree of influence of different brain regions on decision-making is parietal lobe > frontal lobe > occipital lobe > temporal lobe. In addition, the average accuracy of space-CNN reaches 86.13%, which is about 6% higher than similar studies.

Study on the mechanism of nitrapyrin microcapsule suspension effectively improving nitrification inhibition rate in black soil.

Nitrification inhibitors (NIs) have been widely applied to inhibit nitrification and reduce N2O emissions in agriculture. However, there are still some shortcomings, e.g. short effective periods, large applying amounts, low effectiveness, easy deactivation and different effect. Thus, a nitrapyrin microcapsule suspension (CPCS) was used as a new experimental material to elaborate its effects on nitrogen transformation and microbial response mechanisms in black soil by cultivation experiments with six treatments of no fertilization (CK), urea, urea+ 0.2 % CPES, urea+ 0.1 % CPCS, urea+ 0.2 % CPCS, and urea+ 0.3 % CPCS. The content of ammonium, nitrate nitrogen, functional microbial activity, degradation rate and adsorption characteristics of CPCS in the soil at different incubating times were determine. Compared with the nitrapyrin emulsifiable concentrate (CPEC) treatment, the degradation rate of CPCS decreased by 21.54 %, the half-life increased by 10.2 days, and the adsorption rate of nitrapyrin on black soil decreased more than 6-fold. CPCS effectively inhibited the transformation of ammonium nitrogen to nitrate nitrogen within more than 42 days. CPCS had a negative effect on amoA gene abundance and a positive effect on nrfA gene abundance. The research results provide a basic theoretical support for the application of CPCS on black soil.

Integrative preimplantation genetic testing analysis for a Chinese family with hereditary spherocytosis caused by a novel splicing variant of SPTB.

Hereditary spherocytosis (HS), the most common inherited hemolytic anemia disorder, is characterized by osmotically fragile microspherocytic red cells with a reduced surface area on the peripheral blood smear. Pathogenic variants in five erythrocyte membrane structure-related genes ANK1 (Spherocytosis, type 1; MIM#182900), SPTB (Spherocytosis, type 2; MIM#616649), SPTA1 (Spherocytosis, type 3; MIM#270970), SLC4A1 (Spherocytosis, type 4; MIM#612653) and EPB42 (Spherocytosis, type 5; MIM#612690) have been confirmed to be related to HS. There have been many studies on the pathogenic variants and mechanisms of HS, however, studies on how to manage the transmission of HS to the next-generation have not been reported. In this study, we recruited a patient with HS. Targeted next-generation sequencing with a panel of 208 genes related to blood system diseases detected a novel heterozygous variant in the SPTB: c.300+2dup in the proband. Sanger sequencing of variant alleles and haplotype linkage analysis of single nucleotide polymorphism (SNP) based on next-generation sequencing were performed simultaneously. Five embryos were identified with one heterozygous and four not carrying the SPTB variant. Single-cell amplification and whole genome sequencing showed that three embryos had varying degrees of trisomy mosaicism. One of two normal embryos was transferred to the proband. Ultimately, a healthy boy was born, confirmed by noninvasive prenatal testing for monogenic conditions (NIPT-M) to be disease-free. This confirmed our successful application of PGT in preventing transmission of the pathogenic variant allele in the HS family.

Insight of novel biomarkers for papillary thyroid carcinoma through multiomics.

Introduction: The overdiagnosing of papillary thyroid carcinoma (PTC) in China necessitates the development of an evidence-based diagnosis and prognosis strategy in line with precision medicine. A landscape of PTC in Chinese cohorts is needed to provide comprehensiveness. Methods: 6 paired PTC samples were employed for whole-exome sequencing, RNA sequencing, and data-dependent acquisition mass spectrum analysis. Weighted gene co-expression network analysis and protein-protein interactions networks were used to screen for hub genes. Moreover, we verified the hub genes' diagnostic and prognostic potential using online databases. Logistic regression was employed to construct a diagnostic model, and we evaluated its efficacy and specificity based on TCGA-THCA and GEO datasets. Results: The basic multiomics landscape of PTC among local patients were drawn. The similarities and differences were compared between the Chinese cohort and TCGA-THCA cohorts, including the identification of PNPLA5 as a driver gene in addition to BRAF mutation. Besides, we found 572 differentially expressed genes and 79 differentially expressed proteins. Through integrative analysis, we identified 17 hub genes for prognosis and diagnosis of PTC. Four of these genes, ABR, AHNAK2, GPX1, and TPO, were used to construct a diagnostic model with high accuracy, explicitly targeting PTC (AUC=0.969/0.959 in training/test sets). Discussion: Multiomics analysis of the Chinese cohort demonstrated significant distinctions compared to TCGA-THCA cohorts, highlighting the unique genetic characteristics of Chinese individuals with PTC. The novel biomarkers, holding potential for diagnosis and prognosis of PTC, were identified. Furthermore, these biomarkers provide a valuable tool for precise medicine, especially for immunotherapeutic or nanomedicine based cancer therapy.

Novel Intronic Mutations of the SLC12A3 Gene in Patients with Gitelman Syndrome.

Aim: Mutations in the SLC12A3 gene have been reported to cause Gitelman syndrome (GS), characterized by hypokalemic metabolic alkalosis. The aim of this research is to investigate the genetic mutations and clinical features of patients with clinical suspicion of GS. Methods: Six families were enrolled. The symptoms, clinical examination, laboratory results, genotypes, and effect of mutations on mRNA splicing were analyzed. Genomic DNA was screened for gene variations using whole exome sequence and Sanger sequencing. DNA sequences were compared with reference sequences. Results: Genetic analysis revealed nine genetic variants of SLC12A3, including three novel heterozygous mutations (c.1096-2A>G, c.1862A>G, and c.2747+4del) and six previously characterized mutations (c.965-1_976delinsACCGAAAATTTT, c.506-1G>A, c.602-16G>A, c.533C >T, c.1456 G>A, and c.1108 G>C). Probands presented with the clinical syndrome of hypokalemia, increased plasma renin, hypocalciuria and hypokalemic alkalosis. Conclusion: These clinical manifestations and genotypes were consistent with the diagnostic criteria of GS. The study described the phenotypes and genotypes of six pedigrees involving GS patients, demonstrating the importance of SLC12A3 gene screening for GS. This study expands the mutation spectrum of SLC12A3 gene in GS.

Complex Interplay Between Metabolism and CD4+ T-Cell Activation, Differentiation, and Function: a Novel Perspective for Atherosclerosis Immunotherapy.

Atherosclerosis is a complex pathological process that results from the chronic inflammatory reaction of the blood vessel wall and involves various immune cells and cytokines. An imbalance in the proportion and function of the effector CD4+ T-cell (Teff) and regulatory T-cell (Treg) subsets is an important cause of the occurrence and development of atherosclerotic plaques. Teff cells depend on glycolytic metabolism and glutamine catabolic metabolism for energy, while Treg cells mainly rely on fatty acid oxidation (FAO), which is crucial for determining the fate of CD4+ T cells during differentiation and maintaining their respective immune functions. Here, we review recent research achievements in the field of immunometabolism related to CD4+ T cells, focusing on the cellular metabolic pathways and metabolic reprogramming involved in the activation, proliferation, and differentiation of CD4+ T cells. Subsequently, we discuss the important roles of mTOR and AMPK signaling in regulating CD4+ T-cell differentiation. Finally, we evaluated the links between CD4+ T-cell metabolism and atherosclerosis, highlighting the potential of targeted modulation of CD4+ T-cell metabolism in the prevention and treatment of atherosclerosis in the future.

The Loss-Function of KNL1 Causes Oligospermia and Asthenospermia in Mice by Affecting the Assembly and Separation of the Spindle through Flow Cytometry and Immunofluorescence.

KNL1 (kinetochore scaffold 1) has attracted much attention as one of the assembly elements of the outer kinetochore, and the functions of its different domains have been gradually revealed, most of which are associated with cancers, but few links have been made between KNL1 and male fertility. Here, we first linked KNL1 to male reproductive health and the loss-function of KNL1 resulted in oligospermia and asthenospermia in mice (an 86.5% decrease in total sperm number and an 82.4% increase in static sperm number, respectively) through CASA (computer-aided sperm analysis). Moreover, we introduced an ingenious method to pinpoint the abnormal stage in the spermatogenic cycle using flow cytometry combined with immunofluorescence. Results showed that 49.5% haploid sperm was reduced and 53.2% diploid sperm was increased after the function of KNL1 was lost. Spermatocytes arrest was identified at the meiotic prophase I of spermatogenesis, which was induced by the abnormal assembly and separation of the spindle. In conclusion, we established an association between KNL1 and male fertility, providing a guide for future genetic counseling regarding oligospermia and asthenospermia, and a powerful method for further exploring spermatogenic dysfunction by utilizing flow cytometry and immunofluorescence.

A Novel System for the Detection of Spontaneous Abortion-Causing Aneuploidy and Its Erroneous Chromosome Origins through the Combination of Low-Pass Copy Number Variation Sequencing and NGS-Based STR Tests.

During the period of 2018-2020, we first combined reported low-pass whole genome sequencing and NGS-based STR tests for miscarriage samples analysis. Compared with G-banding karyotyping, the system increased the detection rate of chromosomal abnormalities in miscarriage samples to 56.4% in 500 unexplained recurrent spontaneous abortions. In this study, a total of 386 STR loci were developed on twenty-two autosomes and two sex chromosomes (X and Y chromosomes), which can help to distinguish triploidy, uniparental diploidy and maternal cell contamination and can trace the parental origin of erroneous chromosomes. It is not possible to accomplish this with existing methods of detection in miscarriage samples. Among the tested aneuploid errors, the most frequently detected error was trisomy (33.4% in total and 59.9% in the error chromosome group). In the trisomy samples, 94.7% extra chromosomes were of maternal origin and 5.31% were of paternal origin. This novel system improves the genetic analysis method of miscarriage samples and provides more reference information for clinical pregnancy guidance.

A study on and adsorption mechanism of ammonium nitrogen by modified corn straw biochar.

Using corn stover as raw material, the adsorption mechanism of ammonium nitrogen by biochar prepared by different modification methods was studied. The biochar was characterized by Fourier transform infrared spectroscopy, surface-area analysis and scanning electron microscopy. The results showed that the adsorption of NH 4 + - N by different modified biochars confirmed the quasi-second-order kinetic equation (R 2 > 0.95, p ≤ 0.05), the adsorption isotherms of the Langmuir equation (R 2 ≥ 0.96, p ≤ 0.05). ΔG θ < 0, ΔH θ > 0 indicated that the adsorption of NH 4 + - N by different modified biochars was a spontaneous endothermic reaction. With the increase in adsorption temperature, the adsorption capacity of biochar to ammonium nitrogen increased gradually. The adsorption was monolayer adsorption and was controlled by a fast reaction. Both KOH and FeCl3 modified biochars significantly improved the adsorption capacity of NH 4 + - N , and the adsorption mechanism was different. The adsorption capacity of NH 4 + - N by FeCl3 modified biochars mainly increased the specific surface area and micropore volume. The adsorption of ammonium nitrogen after KOH modification primarily depended on the wealthy oxygen-containing functional groups. The adsorption effect of ammonium nitrogen modified by KOH was better than that of biochar modified by FeCl3.

Identification of a Novel Frameshift Variant of ARR3 Related to X-Linked Female-Limited Early-Onset High Myopia and Study on the Effect of X Chromosome Inactivation on the Myopia Severity.

X-linked myopia 26 (Myopia 26, MIM #301010), which is caused by the variants of ARR3 (MIM *301770), is characterized by female-limited early-onset high myopia (eo-HM). Clinical characteristics include a tigroid appearance in the fundus and a temporal crescent of the optic nerve head. At present, the limited literature on eo-HM caused by ARR3 mutations shows that its inheritance mode is complex, which brings certain difficulties to pre-pregnancy genetic counseling, pre-implantation genetic diagnosis, and prenatal diagnosis. Here, we investigated the genetic underpinning of a Chinese family with eo-HM. Whole exome sequencing of the proband revealed a novel frameshift mutation in ARR3 (NM_004312, exon10, c.666delC, p. Asn222LysfsTer22). Although the mode of inheritance of the eo-HM family fits the X-linked pattern of ARR3, the phenotypes of three patients deviate from the typical early-onset high myopia. Through X-chromosome inactivation experiments, the patient's different phenotypes can be precisely explained. In addition, this study not only enhanced the correlation between ARR3 and early-onset high myopia but also provided explanations for different phenotypes, which may inspire follow-up studies. Our results enrich the knowledge of the variant spectrum in ARR3 and provide critical information for preimplantation and prenatal genetic testing, diagnosis, and counseling.

Detection of cryptic balanced chromosomal rearrangements using high-resolution optical genome mapping.

BACKGROUND: Chromosomal rearrangements have profound consequences in diverse human genetic diseases. Currently, the detection of balanced chromosomal rearrangements (BCRs) mainly relies on routine cytogenetic G-banded karyotyping. However, cryptic BCRs are hard to detect by karyotyping, and the risk of miscarriage or delivering abnormal offspring with congenital malformations in carrier couples is significantly increased. In the present study, we aimed to investigate the potential of single-molecule optical genome mapping (OGM) in unravelling cryptic chromosomal rearrangements. METHODS: Eleven couples with normal karyotypes that had abortions/affected offspring with unbalanced rearrangements were enrolled. Ultra-high-molecular-weight DNA was isolated from peripheral blood cells and processed via OGM. The genome assembly was performed followed by variant calling and annotation. Meanwhile, multiple detection strategies, including FISH, long-range-PCR amplicon-based next-generation sequencing and Sanger sequencing were implemented to confirm the results obtained from OGM. RESULTS: High-resolution OGM successfully detected cryptic reciprocal translocation in all recruited couples, which was consistent with the results of FISH and sequencing. All high-confidence cryptic chromosomal translocations detected by OGM were confirmed by sequencing analysis of rearrangement breakpoints. Moreover, OGM revealed additional complex rearrangement events such as inverted aberrations, further refining potential genetic interpretation. CONCLUSION: To the best of our knowledge, this is the first study wherein OGM facilitate the rapid and robust detection of cryptic chromosomal reciprocal translocations in clinical practice. With the excellent performance, our findings suggest that OGM is well qualified as an accurate, comprehensive and first-line method for detecting cryptic BCRs in routine clinical testing.

Modelling maize yield, soil nitrogen balance and organic carbon changes under long-term fertilization in Northeast China.

Optimized fertilization is an effective strategy for improving nitrogen (N) use efficiency and maintaining high crop yield, but its long-term impacts on soil organic carbon (C) and inorganic N dynamics remain unclear. The objectives of this study were to 1) explore the economic optimum N rate and evaluate the DSSAT CERES-Maize model using the measurements from three 3-year maize (Zea mays L.) field experiments, in Gongzhuling and Yushu County, Northeast China, and 2) assess the long-term impacts of farmers' N rate (N250), optimum N rate (N180) and organic-inorganic combined N rate (MN180) on maize yields, soil N and C changes from 1985 to 2020. Results showed that similar maize yields of 8000-11,000 kg ha-1 were achieved under the average economic optimum N rate of 170 kg N ha-1 relative to N250 in both counties. Good agreements were observed between the simulated and measured maize yield, above-ground biomass, N uptake and soil nitrate (NO3--N). Long-term simulation confirmed that N180 and MN180 can achieve the same yield as N250 in both counties. The lowest annual soil inorganic N balance, NO3--N leaching, and nitrous oxide (N2O) and ammonia (NH3) emissions were achieved under MN180, followed by N180 in both sites. Higher NO3--N leaching was found in sandy clay loam soil than silt clay loam and clay loam soils. Average soil organic C (SOC, 0-0.2 m) increased from 1.3 to 2.4% in Gongzhuling and from 2.2 to 2.4% in Yushu under MN180 during the 35-year period, but it showed declining trends under N180 and N250. We concluded that the economic optimum N rate could be an option to replace current farmers' N rate for the continuous maize. Substitution of inorganic fertilizer by 20-30% manure under the optimum N rate showed advantage on maintaining high yield, reducing soil inorganic N losses as well as increasing SOC stock for sustainable agriculture.

A Design and Application of Municipal Service Platform Based on Cloud-Edge Collaboration for Smart Cities.

Information and Communication Technology (ICT) makes cities "smart", capable of providing advanced municipal services to citizens more efficiently. In the literature, many applications of municipal service platform based on cloud computing and edge computing have been proposed, but the reference model and application instance based on cloud-edge collaboration specially for municipal service platform is rarely studied. In this context, this paper first develops a reference model, including resource collaboration, application collaboration, service collaboration, and security collaboration, and discusses the main contents and challenges of each part. Then, aiming at the problem of computing and communication resources allocation in the cloud-edge collaboration, a game-theory-based dynamic resource allocation model is introduced. Finally, an e-government self-service system based on the cloud-edge collaboration is designed and implemented. The cloud side is a cloud computing server, and the edge side are the self-service terminals integrating various edge computing devices with Artificial Intelligence (AI) embedded. The experimental results show that the designed system combines the advantages of cloud computing and edge computing, and provides a better user experience with lower processing latency, larger bandwidth, and more concurrent tasks. Meanwhile, the findings show that the evolutionary equilibrium and the Nash equilibrium are the optimal solutions, respectively.