Advances in miniature CRISPR-Cas proteins and their applications in gene editing.

The CRISPR-Cas system consists of Cas proteins and single-stranded RNAs that recruit Cas proteins and specifically target the nucleic acid. Some Cas proteins can accurately cleave the target nucleic acid under the guidance of the single-stranded RNAs. Due to its exceptionally high specificity, the CRISPR-Cas system is now widely used in various fields such as gene editing, transcription regulation, and molecular diagnosis. However, the huge size of the most frequently utilized Cas proteins (Cas9, Cas12a, and Cas13, which contain 950-1,400 amino acids) can limit their applicability, especially in eukaryotic gene editing, where larger Cas proteins are difficult to deliver into the target cells. Recently discovered miniature CRISPR-Cas proteins, consisting of only 400 to 800 amino acids, offer the possibility of overcoming this limitation. This article systematically reviews the latest research progress of several miniature CRISPR-Cas proteins (Cas12f, Cas12j, Cas12k, and Cas12m) and their practical applications in the field of gene editing.

Pursuing sustainable productivity with millions of smallholder farmers.

Sustainably feeding a growing population is a grand challenge, and one that is particularly difficult in regions that are dominated by smallholder farming. Despite local successes, mobilizing vast smallholder communities with science- and evidence-based management practices to simultaneously address production and pollution problems has been infeasible. Here we report the outcome of concerted efforts in engaging millions of Chinese smallholder farmers to adopt enhanced management practices for greater yield and environmental performance. First, we conducted field trials across China's major agroecological zones to develop locally applicable recommendations using a comprehensive decision-support program. Engaging farmers to adopt those recommendations involved the collaboration of a core network of 1,152 researchers with numerous extension agents and agribusiness personnel. From 2005 to 2015, about 20.9 million farmers in 452 counties adopted enhanced management practices in fields with a total of 37.7 million cumulative hectares over the years. Average yields (maize, rice and wheat) increased by 10.8-11.5%, generating a net grain output of 33 million tonnes (Mt). At the same time, application of nitrogen decreased by 14.7-18.1%, saving 1.2 Mt of nitrogen fertilizers. The increased grain output and decreased nitrogen fertilizer use were equivalent to US$12.2 billion. Estimated reactive nitrogen losses averaged 4.5-4.7 kg nitrogen per Megagram (Mg) with the intervention compared to 6.0-6.4 kg nitrogen per Mg without. Greenhouse gas emissions were 328 kg, 812 kg and 434 kg CO2 equivalent per Mg of maize, rice and wheat produced, respectively, compared to 422 kg, 941 kg and 549 kg CO2 equivalent per Mg without the intervention. On the basis of a large-scale survey (8.6 million farmer participants) and scenario analyses, we further demonstrate the potential impacts of implementing the enhanced management practices on China's food security and sustainability outlook.

Design, synthesis, and biological evaluation of a potent PLK4 inhibitor WY29 with 1H-pyrazolo[3,4-d]pyrimidine scaffold.

Centriole duplication occurs once per cell cycle and is regulated by Polo-like kinase 4 (PLK4). Overexpression of PLK4 in somatic cells can lead to the excessive formation of centrioles, directly causing chromosome segregation errors and tumorigenesis. In this study, we described our efforts to develop a series of PLK4 inhibitors with 1H-pyrazolo[3,4-d]pyrimidine core, and further structure- and receptor-based design and optimization resulted in a potent inhibitor WY29 (IC50 = 0.027 µM), which exhibited good selectivity to other PLK family members (PLK1-3). At the cellular level, compound WY29 showed excellent antiproliferative activity against three breast cancer cell lines (MCF-7, BT474, and MDA-MB-231) while weak inhibitory activity was found on normal cell line HUVECs. In addition, the in vitro preliminary drug-like properties evaluation of compound WY29 showed outstanding stability in human plasma and liver microsomes, and weak inhibitory activity against the major subtypes of human cytochrome P450. Also, the drug-like properties prediction of compound WY29 displayed remarkable drug-like properties (drug-likeness mode score: 1.06). In conclusion, these results support the further development of compound WY29 as a lead compound for PLK4-targeted anticancer drug discovery.

Design, synthesis, and biological evaluation of aminopyridine derivatives as novel tropomyosin receptor kinase inhibitors.

Tropomyosin receptor kinase (TRK) is a successful target for the treatment of various cancers caused by NTRK gene fusions. Herein, based on a rational drug design strategy, we designed and synthesized 35 aminopyrimidine derivatives that were shown to be TRKA inhibitors in the enzyme assay, among which compounds C3, C4, and C6 showed potent inhibitory activities against TRKA with IC50 values of 6.5, 5.0, and 7.0 nM, respectively. In vitro antiproliferative activity study showed that compound C3 significantly inhibited the proliferation of KM-12 cells but had weak inhibitory effect on MCF-7 cells and HUVEC cells. The preliminary druggability evaluation showed that compound C3 exhibited favorable liver microsomal and plasma stabilities and had weak or no inhibitory activity against cytochrome P450 isoforms at 10 µM. Compounds C3, C4, and C6 were also selected for ADME (absorption, distribution, metabolism, and elimination) properties prediction and molecular docking studies. Inhibition experiments showed that compound C3 was not selective for TRK subtypes. All results indicated that compound C3 was a useful candidate for the development of TRK inhibitors.

Amendment with controlled release urea increases leaf morpho-physiological traits, grain yield and NUE in a double-cropping rice system in southern China.

BACKGROUND: Understanding of mechanisms that underpin high-yielding cropping systems is essential for optimizing management practices. Currently, the contribution of plant traits such as leaf area, chlorophyll content and intercepted photosynthetically active radiation (PARi ) to yield and nitrogen use efficiency (NUE) are not fully understood. In addition, the understanding of how canopy traits are affected by nitrogen (N) management practices is unclear. The present study aimed to determine the effect of amendment with controlled release urea (CR), common urea or no urea on NUE and plant eco-physiological characteristics in a 2-year field study in a double rice cropping system. RESULTS: Regulation of N release through amendment with CR significantly increased grain yield, NUE and leaf morpho-physiological attributes. CR coupled with common urea (at comparable total N rates) increased leaf area index (LAI), relative chlorophyll content index (CCI) and PARi , leading to higher grain yield and NUE (increased 24.4% and 25.3% in early and late rice, respectively) compared to local farming practice. Structural equation model (SEM) analysis showed that differences in N application, between CR and common urea, directly accounted for differences observed in soil nutrient, PARi and NUE rather than yield components. Additionally, compared to traditional yield determinants, LAI and PARi (between booting and filling stage) are capable of predicting and explaining grain yield by 0.69 and 0.92 of R2 in early and late rice, respectively. CONCLUSION: Leaf morpho-physiological traits are important for developing N management practices to increase NUE and improve food security for paddy agriculture in southern China. © 2022 Society of Chemical Industry.

High-precision distributed detection of rail defects by tracking the acoustic propagation waves.

Nowadays, early defect detection plays a significant role for the railway safety warning. However, the existing methods cannot satisfy the requirements of real-time and high-precision detection. Here, a high-precision, distributed and on-line method for detecting rail defect is proposed and demonstrated. When a train goes through defects, the instantaneous elastic waves will be excited by the wheel-rail interaction, which will further propagate along railway tracks bidirectionally. Through mounting the backscattering enhanced optical fiber on the railway as sensors, the fiber optic distributed acoustic sensing system can record the propagation trace precisely. Further, the acoustic propagation fitting method is applied onto the propagation data to detect and locate defects along the long-distance railway. Especially, the dual-frequency joint-processing algorithm is proposed to improve the location accuracy. The field test proves that multiple defects along the railway can be successfully identified and located with a standard deviation of 0.314m. To the best of our knowledge, this work is the first report of distributed rail defect detection, which will bring a breakthrough for high-precision structural damage detection in the infrastructures such as the railway, pipeline and tunnel.

Therapeutic potential of targeting LSD1/ KDM1A in cancers.

LSD1 was the first histone demethylase identified by Professor Shi Yang and his team members in 2004. LSD1 employs FAD as its cofactor, which catalyzes the demethylation of H3K4 and H3K9. It is aberrantly overexpressed in different types of cancers and is associated with the growth, invasion, and metastasis of cancer cells. The knockout or inhibition of LSD1 could effectively suppress tumor development, and thus, it has become an attractive molecular target for cancer therapy. Moreover, many LSD1 inhibitors have been developed in preclinical and clinical trials to treat solid tumors and hematological malignancy. This study made an extensive review of the research obtained from the literature retrieval of electronic databases, such as PubMed, Web of Science, RCSB PDB, ClinicalTrials.gov, and EU clinical trials register. This review summarizes recent studies on the advances of LSD1 inhibitors in the literature, covering January 2015 to June 2021. It focuses on the function of LSD1 in tumor cells, summarizes the crystal structures of Homo sapiens LSD1, reviews the structural characteristics of LSD1 inhibitors, compares the screening methods of LSD1 inhibitors, and proposes guidelines for the future exploitation of LSD1 inhibitors.

Design, synthesis, and biological evaluation of potent FAK-degrading PROTACs.

FAK mediated tumour cell migration, invasion, survival, proliferation and regulation of tumour stem cells through its kinase-dependent enzymatic functions and kinase-independent scaffolding functions. At present, the development of FAK PROTACs has become one of the hotspots in current pharmaceutical research to solve above problems. Herein, we designed and synthesised a series of FAK-targeting PROTACs consisted of PF-562271 derivative 1 and Pomalidomide. All compounds showed significant in vitro FAK kinase inhibitory activity, the IC50 value of the optimised PROTAC A13 was 26.4 nM. Further, A13 exhibited optimal protein degradation (85% degradation at 10 nM). Meantime, compared with PF-562271, PROTAC A13 exhibited better antiproliferative activity and anti-invasion ability in A549 cells. More, A13 had excellent plasma stability with T1/2 >194.8 min. There are various signs that PROTAC A13 could be useful as expand tool for studying functions of FAK in biological system and as potential therapeutic agents.

Design, synthesis, and biological evaluation of coumarin analogs as novel LSD1 inhibitors.

The abnormal expression of lysine-specific histone demethylase 1 (LSD1) is associated with different cancer types, and it is increasingly recognized as a potential therapeutic target in oncology. Here, utilizing core hopping and conformational restriction strategies, we designed and synthesized a series of coumarin analogs that were shown to be potent LSD1 inhibitors in the enzyme assay. Furthermore, several potent compounds were selected to evaluate their antiproliferative activity on A549 cells and MGC-803 cells with high expression of LSD1. Among them, YX10 showed an anticlonogenic effect on A549 cells and MGC-803 cells, with IC50 values of 1.52 ± 0.16 and 0.98 ± 0.18 µM, respectively. Modeling suggested that the inhibitors would bind to the active site of the protein located around the key residues of Asp555 and Lys661. Meanwhile, a preliminary druggability evaluation showed that compound YX10 showed favorable liver microsomal and moderate plasma stability and weak inhibitory activity against cytochrome P450 isoforms at 10 µM. All the results indicated that compound YX10 could represent a promising lead compound for further development.

Design, synthesis, and evaluation of novel 3,4-isoxazolediamide derivatives for the combination treatment of azole-resistant candidiasis.

Invasive fungal infections are emerging as serious infectious diseases worldwide. Due to the frequent emergence of resistance, the cure for invasive fungal infections is often unachievable. The molecular chaperone Hsp90 provides a promising target because it supports survival, virulence, and drug resistance in a variety of pathogens. Herein, we report on the structural optimization and structure-activity relationship studies of 3,4-isoxazolediamide analogs. As a new class of fungal Hsp90 inhibitor, compound B25 was found to have good synergistic effects with fluconazole and to avoid potential mammalian toxicity. It also showed remarkable metabolic stability in vitro. Collectively, B25 could be a promising lead compound for drug discovery targeting fungal Hsp90 and deserves further investigation.

Design, synthesis, and biological evaluation of 5-aminotetrahydroquinoline-based LSD1 inhibitors acting on Asp375.

The abnormal expression of lysine-specific histone demethylase 1 (LSD1) is associated with different cancer types, and LSD1 inhibitory activity seems to have high therapeutic potential in cancer treatment. Here, we report the design, synthesis, and biochemical evaluation of novel 5-aminotetrahydroquinoline-based LSD1 inhibitors. Among them, compounds A6, A8, B1-B5, and C4 showed preferable inhibitory effects on LSD1, with IC50 = 0.19-0.82 µM. Several potent compounds were selected to evaluate their antiproliferative activity on A549 cells and MCF-7 cells with a high expression of LSD1. The potential binding modes of the compounds were revealed through molecular docking to rationalize the potency of compounds toward LSD1. Our data recognized that the 5-aminotetrahydroquinoline scaffold may serve as a starting point for developing potent LSD1 inhibitors for cancer therapy.

Producing more grain with lower environmental costs.

Agriculture faces great challenges to ensure global food security by increasing yields while reducing environmental costs. Here we address this challenge by conducting a total of 153 site-year field experiments covering the main agro-ecological areas for rice, wheat and maize production in China. A set of integrated soil-crop system management practices based on a modern understanding of crop ecophysiology and soil biogeochemistry increases average yields for rice, wheat and maize from 7.2 million grams per hectare (Mg ha(-1)), 7.2 Mg ha(-1) and 10.5 Mg ha(-1) to 8.5 Mg ha(-1), 8.9 Mg ha(-1) and 14.2 Mg ha(-1), respectively, without any increase in nitrogen fertilizer. Model simulation and life-cycle assessment show that reactive nitrogen losses and greenhouse gas emissions are reduced substantially by integrated soil-crop system management. If farmers in China could achieve average grain yields equivalent to 80% of this treatment by 2030, over the same planting area as in 2012, total production of rice, wheat and maize in China would be more than enough to meet the demand for direct human consumption and a substantially increased demand for animal feed, while decreasing the environmental costs of intensive agriculture.

Discovery of 2,4-diaminopyrimidine derivatives targeting p21-activated kinase 4: Biological evaluation and docking studies.

In this study, novel 2,4-diaminopyrimidine derivatives targeting p21-activated kinase 4 (PAK4) were discovered and evaluated for their biological activity against PAK4. Among the derivatives studied, promising compounds A2, B6, and B8 displayed the highest inhibitory activities against PAK4 (IC50 = 18.4, 5.9, and 20.4 nM, respectively). From the cellular assay, compound B6 exhibited the highest potency with an IC50 value of 2.533 µM against A549 cells. Some compounds were selected for computational ADME (absorption, distribution, metabolism, and elimination) properties and molecular docking studies against PAK4. The detailed structure-activity relationship based on the biochemical activities and molecular docking studies were explored. According to the docking studies, compound B6 had the lowest docking score (docking energy: -7.593 kcal/mol). The molecular docking simulation indicated the binding mode between compound B6 and PAK4. All these results suggest compound B6 as a useful candidate for the development of a PAK4 inhibitor.

Integrated systematic approach increase greenhouse tomato yield and reduce environmental losses.

High input - high output greenhouse vegetable systems are responsible for nutrient surpluses and environmental losses. Integrated strategies that improve soil, crop and nutrient management are needed to ensure more sustainable production systems. We conducted a two-year field experiment to evaluate the potential of integrated soil-crop system management (ISSM) practices to improve the productivity and environmental outcomes from an intensive greenhouse tomato production system in the Yangtze River Basin, China. Four treatments were tested: i) farmers' practice (FP); ii) soil remediation (SR), where lime nitrogen with compost addition was the only management strategy; iii) a treatment that combined soil remediation with optimized crop planting density (SRCO), which increased planting density for improving crop yield; and iv) integrated soil-crop system management (ISSM), as a systematic integrated approach, which included the combined optimization of soil remediation, crop optimization, and nutrient management. In the integrated soil-crop system management treatment, nutrient management was optimized through adoption of the most appropriate type (formula) of fertilizer for the crop, rate and application timing of synthetic fertilizer, and by substituting poultry manure with compost. Our results indicated that the fruit yield of the integrated soil-crop system management treatment was 104 t ha-1, 13.4%-37.3% higher than that of the other three treatments. The mean reactive nitrogen loss (81.1 kg N ha-1) and the greenhouse gas emissions (6495 kg CO2-eq ha-1) in the farmers' practice treatment were much higher than in the other three treatments (reactive nitrogen loss: 47.9-54.3 kg N ha-1; and greenhouse gas emissions: 4926-5468 kg CO2-eq ha-1, respectively). The mean nitrogen and carbon footprints of the integrated soil-crop system management treatment were significantly lower than those of other treatments, as a result of both the lower fertilizer nitrogen use and the greater yield. This study indicates that integrated soil-crop system management could produce greater yields and increase net profit with reduced nitrogen inputs, whilst reducing the environmental cost associated with conventional farmers' practice in plastic-greenhouse vegetable production systems.

Impact of biological manure substitution on grain yield, nitrogen recovery efficiency, and soil biochemical properties.

Fertilization plays a crucial role in ensuring global food security and ecological balance. This study investigated the impact of substituting innovative biological manure for chemical fertilization on rice (Oryza sativa L) productivity and soil biochemical properties based on a three-year experiment. Our results suggested rice yield and straw weight were increased under manure addition treatment. Specifically, 70% of total nitrogen (N) fertilizer substituted by biological manure derived from straw, animal waste and microbiome, led to a substantial 13.6% increase in rice yield and a remarkable 34.2% boost in straw weight. In comparison to the conventional local farmer practice of applying 165 kg N ha-1, adopting 70% of total N plus biological manure demonstrated superior outcomes, particularly in enhancing yield components and spike morphology. Fertilization treatments led to elevated levels of soil microbial biomass carbon and N. However, a nuanced comparison with local practices indicated that applying biological manure alongside urea resulted in a slight reduction in N content in vegetative and economic organs, along with decreases of 10.4%, 11.2%, and 6.1% in N recovery efficiency (NRE), respectively. Prudent N management through the judicious application of partial biological manure fertilizer in rice systems could be imperative for sustaining productivity and soil fertility in southern China.

Discovery of novel 3-(1H-pyrazol-4-yl)-1H-indazole derivatives as potent type II TRK inhibitors against acquired resistance.

Tropomyosin receptor kinase (TRK) is a promising target for treating NTRK fusion cancers. The solvent front and xDFG mutations induced by larotrectinib and entrectinib result in acquired resistance in advanced-stage patients. In this study, we report a highly potent and selective type II TRK inhibitor, 40l, developed using a structure-based design strategy. Compound 40l significantly suppressed Km-12, Ba/F3-TRKAG595R, and Ba/F3-TRKAG667C cell proliferation. In biochemical and cellular assays, 40l showed better inhibitory activity against TRKAG667C than that by the positive control, selitrectinib. Additionally, it induced apoptosis of Ba/F3-TRKAG595R and Ba/F3-TRKAG667C cells in a dose-dependent manner. Furthermore, 40l showed good selectivity for a panel of 41 kinases. In vitro assays indicated that 40l possessed outstanding plasma stability and moderate liver microsomal stability. Based on the above results, compound 40l could be further optimized to overcome the solvent front and xDFG TRK mutations.

Blended controlled-release nitrogen fertilizer increases rice post-anthesis nitrogen accumulation, translocation and nitrogen-use efficiency.

One-time application of blended controlled-release nitrogen fertilizer (CRN) has the potential to solve the difficulty of top-dressing fertilizer in the cultivation of rice and reduce the cost of CRN fertilizer application. However, its effects on rice dry matter and nitrogen (N) accumulation and translocation, yield and N-use efficiency (NUE) remain uncertain. Field experiments were carried out at three sites (Mingguang, Chaohu, and Guichi) in the Yangtze River Delta in China to compare the effects of the conventional split applications of urea and the blended CRN and on post-anthesis dry matter and N accumulation and translocation, yield, and NUE in rice at 0, 60, 120, 180, and 240 kg N ha-1. The results showed that at the equal N application rates, compared under the conventional N fertilizer treatment, the blended CRN application significantly increased the rice yield by an average of 0.9-6.9%, mainly due to increase the number of spikelets per panicle. The highest yield achieved with blended CRN treatment occurred at 200 kg N ha-1, with an NUE of 45.9%. Moreover, in comparison to the conventional N fertilizer, the blended CRN treatment increased pre-anthesis N translocation (Pre-NT) by 1.0-19.8%, and the contribution of pre-NT to grain N by 0.2-8.7%, and NUE by 3.2-28.4%. Meanwhile, the blended CRN treatment reduced labor costs by 1800 Yuan ha-1 and enhanced the economic gains by 21.5-68.8%. Therefore, one-time application of blended CRN ≤ 200 kg N ha-1 application rate improved rice yield, NUE, and economic profit compared to equivalent rates of split applied conventional N fertilizers.

Design, synthesis and evaluation of 2-phenylpyrimidine derivatives as novel antifungal agents targeting CYP51.

Invasive fungal infections, with high morbidity and mortality, have become one of the most serious threats to human health. There are a few kinds of clinical antifungal drugs but large amounts of them are used, so there is an urgent need for a new structural type of antifungal drug. In this study, we carried out three rounds of structural optimisation and modification of the compound YW-01, which was obtained from the preliminary screening of the group, by using the strategy of scaffold hopping. A series of novel phenylpyrimidine CYP51 inhibitors were designed and synthesised. In vitro antifungal testing showed that target compound C6 exhibited good efficacy against seven common clinically susceptible strains, which was significantly superior to the clinical first-line drug fluconazole. Subsequently in vitro tests on metabolic stability and cytotoxicity revealed that C6 was safe and stable for hepatic microsomal function. Finally, C6 warranted further exploration as a possible novel structural type of CYP51 inhibitor.

Effects of Partial Replacement of Nitrogen Fertilizer with Organic Fertilizer on Rice Growth, Nitrogen Utilization Efficiency and Soil Properties in the Yangtze River Basin.

Cake fertilizer and dairy manure were used as experimental materials to carry out a 9-year (2012-2020) field experiment in the main rice production areas in the Yangtze River basin. Different fertilization modes were used (no fertilization, CK; chemical fertilizer application alone, HY; reduced fertilization with chemical fertilizer application, RF; cake fertilizer replacement of nitrogen fertilizer, CFR; and dairy manure replacement of nitrogen fertilizer, DMR). Changes in the total rice yield, yield components, absorption of nitrogen, soil pH, organic matter, total nitrogen, and soil bulk density under different fertilization treatments were analyzed. The results show that organic fertilizer replacement leads to a stable and high rice yield. The 9-year average rice yields of the CFR and DMR treatments were 60.0% and 61.5% higher than that of CK. The nitrogen uptake of the CFR and DMR treatments was also higher than that of the other treatments. The nitrogen recovery efficiency in the current season could be increased by 16.37-22.89%, and after 9 years of testing, the soil total nitrogen contents of CFR and DMR increased by 0.23-0.85 g·kg-1 compared to the other treatments. The available P and K contents of DMR increased by 30.17 mg·kg-1 and 22.02 mg·kg-1 compared with HY, respectively. The soil bulk density was reduced by 0.08 g·cm-3. Generally, the effects of dairy manure replacement were better than those of cake fertilizer. This is an important method that can be used to fertilize the soil and foster sustainable soil utilization in the rice-growing area of the Yangtze River Basin, as a long-term partial replacement for chemical nitrogen fertilizer.

The optimum economic nitrogen rate of blended controlled-release nitrogen fertilizer for rice in the Chanoyu watershed in the Yangtze River Delta, China.

Introduction: The application of controlled-release nitrogen fertilizer (CRN) has become an important production method to achieve high crop yield and ecological safety. However, the rate of urea-blended CRN for rice is usually determined by conventional urea, and the actual rate is still unclear. Methods: A five-year field experiment was carried out in the Chaohu watershed in the Yangtze River Delta to study rice yield, N fertilizer utilization efficiency (NUE), ammonia (NH3) volatilization and economic benefit under the four urea-blended CRN treatments with a 4:3:3 ratio applied at one time (60, 120, 180, 240 kg/hm2, CRN60, CRN120, CRN180, CRN240), four conventional N fertilizer treatments (N60, N120, N180, N240) and a control without N fertilizer (N0). Results and Discussion: The results showed that the N released from the blended CRNs could well satisfy the N demand of rice growth. Similar to the conventional N fertilizer treatments, a quadratic equation was used to model the relationship between rice yield and N rate under the blended CRN treatments. The blended CRN treatments increased rice yield by 0.9-8.2% and NUE by 6.9-14.8%, respectively, compared with the conventional N fertilizer treatments at the same N application rate. The increase in NUE in response to applied blended CRN was related to the reduction in NH3 volatilization. Based on the quadratic equation, the five-year average NUE under the blended CRN treatment was 42.0% when rice yield reached the maximum, which was 28.9% higher than that under the conventional N fertilizer treatment. Among all treatments, CRN180 had the highest yield and net benefit in 2019. Considering the yield output, environmental loss, labor and fertilizer costs, the optimum economic N rate under the blended CRN treatment in the Chaohu watershed was 180-214 kg/hm2, compared with 212-278 kg/hm2 under the conventional N fertilizer treatment. The findings suggest that blended CRN improved rice yield, NUE and economic income while decreasing NH3 volatilization and negative environmental outcomes.