Discovery and Optimization of Potent, Efficacious and Selective Inhibitors Targeting EGFR Exon20 Insertion Mutations.

Herein, we report the identification and optimization of a series of potent inhibitors of EGFR Exon20 insertions with significant selectivity over wild-type EGFR. A strategically designed HTS campaign, multiple iterations of structure-based drug design (SBDD), and tactical linker replacement led to a potent and wild-type selective series of molecules and ultimately the discovery of 36. Compound 36 is a potent and selective inhibitor of EGFR Exon20 insertions and has demonstrated encouraging efficacy in NSCLC EGFR CRISPR-engineered H2073 xenografts that carry an SVD Exon20 insertion and reduced efficacy in a H2073 wild-type EGFR xenograft model compared to CLN-081 (5), indicating that 36 may have lower EGFR wild-type associated toxicity.

Identification of PANoptosis-related predictors for prognosis and tumor microenvironment by multiomics analysis in glioma.

PANoptosis is a newly described inflammatory programmed cell death, that highlights coordination between pyroptosis, apoptosis and necroptosis. However, the functions of PANoptosis-related genes in glioma progression still remain to be explored. This study aims to identify PANoptosis-related predictors that may be utilized for prognosis prediction and development of new therapeutic targets. Firstly, bulk and single-cell RNA-seq (scRNA-seq) data of glioma patients were extracted from TCGA, CGGA and GEO database. Genetic analysis indicates a considerably high mutation frequency of PANoptosis-related genes (PANRGs) in glioma. Consensus clustering was applied to reveal different subtypes of glioma based on PANRGs. Two PANoptosis subtypes with distinct prognostic and TME characteristics were identified. Then, with LASSO-Cox regression analysis, four PANoptosis-related predictors (MYBL2, TUBA1C, C21orf62 and KCNIP2) were determined from bulk and scRNA-seq analysis. Predictive PANRG score model was established with these predictors and its correlation with tumor microenvironment (TME) was investigated. The results showed that patients with low PANRG score, had higher infiltration of anti-tumor immune cells, higher MSI score and lower TIDE score, which are more likely to benefit from immunotherapy. Further analysis identified 16 potential drugs associated with PANoptosis-related predictors. Moreover, the expression levels of four PANoptosis-related predictors were examined in clinical samples and the results were consistent with those analyzed in the database. Besides, we also confirmed the biological functions of two oncogenic predictors (MYBL2 and TUBA1C) by cell experiments, which revealed that knockdown of MYBL2 or TUBA1C could significantly inhibit the proliferation and migration of glioma cells. These findings highlight the prognostic value and biological functions of PANRGs in glioma, which may provide valuable insights for individualized treatment.

Whole transcriptome sequencing identifies a competitive endogenous RNA network that regulates the immunity of bladder cancer.

Several types of non-coding RNAs such as circRNAs, lncRNAs, and miRNAs have been identified to regulate mRNAs through the mechanism known as the competitive endogenous RNA (ceRNA) network. To explore the role of the ceRNA regulatory network in the immune microenvironment of bladder cancer, whole-transcriptome sequencing of bladder tumor and its peritumoral tissues from 38 bladder cancer patients, with a total of 63 samples, was performed to screen differentially expressed circ-, lnc-, mi-, and mRNAs to construct a circ/lnc-mi-mRNA regulatory network with pruning algorithms. We excavated a key immune-related gene BDNF to build the final ceRNA network as hsa-miR-107 sponged by hsa-circ-000211, AC108488.1, and LINC00163. Finally, a meta-analysis of 7 public datasets demonstrated that low expression of BDNF and high expression of hsa-miR-107 were associated with longer survival. Our study identified a ceRNA regulatory network as a potentially new prognostic marker and molecular therapeutic target of bladder cancer.

Large language models assisted multi-effect variants mining on cerebral cavernous malformation familial whole genome sequencing.

Cerebral cavernous malformation (CCM) is a polygenic disease with intricate genetic interactions contributing to quantitative pathogenesis across multiple factors. The principal pathogenic genes of CCM, specifically KRIT1, CCM2, and PDCD10, have been reported, accompanied by a growing wealth of genetic data related to mutations. Furthermore, numerous other molecules associated with CCM have been unearthed. However, tackling such massive volumes of unstructured data remains challenging until the advent of advanced large language models. In this study, we developed an automated analytical pipeline specialized in single nucleotide variants (SNVs) related biomedical text analysis called BRLM. To facilitate this, BioBERT was employed to vectorize the rich information of SNVs, while a deep residue network was used to discriminate the classes of the SNVs. BRLM was initially constructed on mutations from 12 different types of TCGA cancers, achieving an accuracy exceeding 99%. It was further examined for CCM mutations in familial sequencing data analysis, highlighting an upstream master regulator gene fibroblast growth factor 1 (FGF1). With multi-omics characterization and validation in biological function, FGF1 demonstrated to play a significant role in the development of CCMs, which proved the effectiveness of our model. The BRLM web server is available at http://1.117.230.196.

Urea Coated with Polyaspartic Acid-Chitosan Increases Foxtail Millet (Setaria italica L. Beauv.) Grain Yield by Improving Nitrogen Metabolism.

Innovative measures of nitrogen (N) fertilization to increase season-long N availability is essential for gaining the optimal foxtail millet (Setaria italica L. Beauv.) productivity and N use efficiency. A split plot field experiment was conducted using the foxtail millet variety Huayougu 9 in 2020 and 2021 in Northeast China to clarify the physiological mechanism of a novel polyaspartic acid-chitosan (PAC)-coated urea on N assimilation and utilization from foxtail millet. Conventional N fertilizer (CN) and the urea-coated -PAC treatments were tested under six nitrogen fertilizer application levels of 0, 75, 112.5, 150, 225, and 337.5 kg N ha-1. The results showed that compared to CN, PN increased the foxtail millet yield by 5.53-15.75% and 10.43-16.17% in 2020 and 2021, respectively. PN increased the leaf area index and dry matter accumulation by 7.81-18.15% and 12.91-41.92%, respectively. PN also enhanced the activities of nitrate reductase, glutamine synthetase, glutamic oxaloacetic transaminase, and glutamic-pyruvic transaminase, thereby increasing the soluble protein in the leaf, plant, and grain N content at harvest compared to CN. Consequently, partial factor productivity from applied N, the agronomic efficiency of applied N, recovery efficiency of applied N, and physiological efficiency of applied N of foxtail millet under PN treatments compared to CN were increased. The improvement effect of the items above was more noticeable under the low-middle N application levels (75, 112.5, and 150 kg N ha-1). In conclusion, the PAC could achieve the goal of high yield and high N use efficiency in foxtail millet under the background of a one-time basic fertilizer application.

Green synthesis of iron nanoparticles using mulberry leaf extract: characterization, identification of active biomolecules, and catalytic activity.

The synthesis of iron-based nanoparticles (Fe NPs) using traditional preparation methods suffered from the disadvantages of high cost, environmental harm, and easy agglomeration. In this study, a novel eco-friendly method was proposed for the synthesis of iron nanomaterials (ML-Fe NPs): using antioxidant components extracted from mulberry leaf to reduce divalent iron (II). The preparation conditions of ML-Fe NPs were optimized by orthogonal tests. The prepared ML-Fe NPs exhibited an amorphous core-shell structure, displaying excellent dispersion and stability. During the synthesis process of ML-Fe NPs, the polyphenol molecules in mulberry leaf extract played a dominant role. A possible synthetic mechanism involving complexation, reduction, and encapsulation was proposed. Furthermore, the ML-Fe NPs were utilized to construct an ML-Fe NPs/peroxymonosulfate catalytic system for the degradation of Rhodamine B dye wastewater. The ML-Fe NPs demonstrated remarkable catalytic potential, achieving a 99% degradation efficiency for Rhodamine B within a span of 40 min.

FOXO3a-interacting proteins' involvement in cancer: a review.

Due to its role in apoptosis, differentiation, cell cycle arrest, and DNA damage repair in stress responses (oxidative stress, hypoxia, chemotherapeutic drugs, and UV irradiation or radiotherapy), FOXO3a is considered a key tumor suppressor that determines radiotherapeutic and chemotherapeutic responses in cancer cells. Mutations in the FOXO3a gene are rare, even in cancer cells. Post-translational regulations are the main mechanisms for inactivating FOXO3a. The subcellular localization, stability, transcriptional activity, and DNA binding affinity for FOXO3a can be modulated via various post-translational modifications, including phosphorylation, acetylation, and interactions with other transcriptional factors or regulators. This review summarizes how proteins that interact with FOXO3a engage in cancer progression.

Establishment of a prognosis prediction model for lung squamous cell carcinoma related to PET/CT: basing on immunogenic cell death-related lncRNA.

BACKGROUND: Immunogenic cell death (ICD) stimulates adaptive immunity and holds significant promise in cancer therapy. Nevertheless, the influence of ICD-associated long non-coding RNAs (lncRNAs) on the prognosis of patients with lung squamous cell carcinoma (LUSC) remains unexplored. METHODS: We employed data from the The Cancer Genome Atlas (TCGA)database to identify ICD-related lncRNAs associated with the prognosis of LUSC using univariate Cox regression analysis. Subsequently, we utilized the LOSS regression model to construct a predictive risk model for assessing the prognosis of LUSC patients based on ICD-related lncRNAs. Our study randomly allocated187 TCGA patients into a training group and 184 patients for testing the predictive model. Furthermore, we conducted quantitative polymerase chain reaction (qPCR) analysis on 43 tumor tissues from LUSC patients to evaluate lncRNA expression levelsPearson correlation analysis was utilized to analyze the correlation of risk scores with positron emission tomography/computed tomography (PET/CT) parameters among LUSC patients. RESULTS: The findings from the univariate Cox regression revealed 16 ICD-associated lncRNAs linked to LUSC prognosis, with 12 of these lncRNAs integrated into our risk model utilizing the LOSS regression. Survival analysis indicated a markedly higher overall survival time among patients in the low-risk group compared to those in the high-risk group. The area under the Receiver operating characteristic (ROC) curve to differentiate high-risk and low-risk patients was 0.688. Additionally, the overall survival rate was superior in the low-risk group compared to the high-risk group. Correlation analysis demonstrated a positive association between the risk score calculated based on the ICD-lncRNA risk model and the maximum standard uptake value (SUVmax) (r = 0.427, P = 0.0043) as well as metabolic volume (MTV)of PET-CT (r = 0.360, P = 0.0177) in 43 LUSC patients. CONCLUSION: We have successfully developed a risk model founded on ICD-related lncRNAs that proves effective in predicting the overall survival of LUSC patients.

Research on integrated inventory transportation optimization of inbound logistics via a VMI-TPL model of an existing enterprise.

Third-party logistics companies face a challenging task in minimizing inventory transportation costs due to the complexities of managing numerous suppliers. Effectively optimizing costs becomes a formidable problem for such companies. This empirical research has yielded strategies for minimizing the inventory transportation cost specifically for company D. Through a rigorous optimization process, the findings presented in this paper demonstrate an average reduction of 7.18% in company D's inventory transportation cost. By jointly optimizing inbound logistics inventory transportation under VMI-TPL mode, this study extends the theory of supplier managed inventory and improves the inbound logistics mode. The results of this study can provide quantitative support and decision-making references for the project operation management of company D and similar enterprises.

Dynamic RBM47 ISGylation confers broad immunoprotection against lung injury and tumorigenesis via TSC22D3 downregulation.

ISGylation is a well-established antiviral mechanism, but its specific function in immune and tissue homeostasis regulation remains elusive. Here, we reveal that the RNA-binding protein RBM47 undergoes phosphorylation-dependent ISGylation at lysine 329 to regulate immune activation and maintain lung homeostasis. K329R knockin (KI) mice with defective RBM47-ISGylation display heightened susceptibility to LPS-induced acute lung injury and lung tumorigenesis, accompanied with multifaceted immunosuppression characterized by elevated pro-inflammatory factors, reduced IFNs/related chemokines, increased myeloid-derived suppressor cells, and impaired tertiary lymphoid structures. Mechanistically, RBM47-ISGylation regulation of the expression of TSC22D3 mRNA, a glucocorticoid-inducible transcription factor, partially accounts for the effects of RBM47-ISGylation deficiency due to its broad immunosuppressive activity. We further demonstrate the direct inhibitory effect of RBM47-ISGylation on TSC22D3 expression in human cells using a nanobody-targeted E3 ligase to induce site-specific ISGylation. Furthermore, epinephrine-induced S309 phosphorylation primes RBM47-ISGylation, with epinephrine treatment exacerbating dysregulated cytokine expression and ALI induction in K329R KI mice. Our findings provide mechanistic insights into the dynamic regulation of RBM47-ISGylation in supporting immune activation and maintaining lung homeostasis.

Overexpression of Glutathione S-Transferases in Human Diseases: Drug Targets and Therapeutic Implications.

Glutathione S-transferases (GSTs) are a major class of phase II metabolic enzymes. Besides their essential role in detoxification, GSTs also exert diverse biological activities in the occurrence and development of various diseases. In the past few decades, much research interest has been paid to exploring the mechanisms of GST overexpression in tumor drug resistance. Correspondingly, many GST inhibitors have been developed and applied, solely or in combination with chemotherapeutic drugs, for the treatment of multi-drug resistant tumors. Moreover, novel roles of GSTs in other diseases, such as pulmonary fibrosis and neurodegenerative diseases, have been recognized in recent years, although the exact regulatory mechanisms remain to be elucidated. This review, firstly summarizes the roles of GSTs and their overexpression in the above-mentioned diseases with emphasis on the modulation of cell signaling pathways and protein functions. Secondly, specific GST inhibitors currently in pre-clinical development and in clinical stages are inventoried. Lastly, applications of GST inhibitors in targeting cell signaling pathways and intracellular biological processes are discussed, and the potential for disease treatment is prospected. Taken together, this review is expected to provide new insights into the interconnection between GST overexpression and human diseases, which may assist future drug discovery targeting GSTs.

False lumen-dependent segmental arteries are associated with spinal cord injury in frozen elephant trunk procedure for acute type I aortic dissection.

Objective: To investigate the association between false lumen (FL) dependency of segmental arteries (SAs) at T9-L3 levels and the risk of spinal cord injury (SCI) following total arch replacement and frozen elephant trunk (FET) implantation in the setting of acute DeBakey type I aortic dissection (AAD). Methods: The study involved consecutive patients with AAD who underwent total arch replacement and FET implantation between 2020 and 2022. Primary outcome was postoperative SCI. The inverse probability of treatment weighting (IPTW) method was employed to minimize the impact of no-randomization bias. Antegrade placement of FET was followed by end-to-end anastomosis of a 4-branch arch graft at the proximal landing site of FET. Results: A total of 146 patients were included (age, 50.5 ± 11.7 years, 115 male), of whom 35 (24%) had SAs at T9-L3 levels completely dependent on FL (FL-dependency group). There was no significant difference in early (30-day or in-hospital) mortality rates between FL-dependency (14.3%) and FL-independency (18.0%) groups (P = .80), however, the rate of SCI was significantly higher in the FL-Dependency group (34.3% vs 2.7%, P < .001). After adjustments, FL dependency was associated with a significantly increased risk of SCI (odds ratio, 13.1; 95% confidence interval, 4.2-41.0; P < .001), whereas it was not significantly associated with risks of early mortality or other major complications (P = .16-.98). Conclusions: FL dependency of SAs at the T9-L3 levels was significantly associated with the development of SCI following FET implantation in AAD, warning against its uses on patients presenting with FL dependency of SAs at critical segments.

Effects of gibberellic acid, kinetin and indole butyric acid mixture on sorghum salinity tolerance and grain yield in saline-alkali coastal zone.

The development and utilization of coastal saline-alkali lands hold significant importance in mitigating the shortage of cultivated land resources in China, enhancing the agro-ecological environment in coastal saline and alkaline areas, and ensuring national food security. We set up both pot and field trials (randomized block design) at Xinxiang experimental station of Institute of Crop Science, Chinese Academy of Agricultural Sciences (ICS-CAAS) and Dongying Yellow River Delta Modern Agricultural Research Base in Shandong Province in 2021 and 2022, respectively. The experimental materials, Jiliang 1 and Jiliang 2, underwent seed dressing with GKI composites at concentrations of 2.5 and 5 mL·kg-1. These composites, which contained the main components of gibberellin, kinetin, and indole butyric acid, were denoted as GKI2.5 and GKI5.0, respectively. The control plots (CK) received water seed dressing. The aim was to assess the regulatory effects of GKI on salt tolerance and grain sorghum yield. Compared to CK, the GKI2.5 and GKI5.0 seed dressing treatments significantly enhanced the growth and development of the two grain sorghum varieties, increased antioxidant enzyme activity and soluble protein content of sorghum leaves, while reducing leaf malondialdehyde content. Moreover, the GKI treatments increased leaf net photosynthetic rate. Under field conditions, yields of Jiliang 1 and Jiliang 2 were enhanced by an average of 17.1% and 19.1%, respectively. In conclusion, GKI seed dressing treatment effectively promoted the growth and development of sorghum under salt stress. It enhanced the antioxidant and osmoregulatory capacities of leaves, reduced the level of membrane lipid peroxidation, and improved net photosynthetic rate of leaves, which together improved the salt tolerance and sorghum yield.

METTL3 exerts synergistic effects on m6A methylation and histone modification to regulate the function of VGF in lung adenocarcinoma.

BACKGROUND: Multiple genetic and epigenetic regulatory mechanisms play a vital role in tumorigenesis and development. Understanding the interplay between different epigenetic modifications and its contribution to transcriptional regulation in cancer is essential for precision medicine. Here, we aimed to investigate the interplay between N6-methyladenosine (m6A) modifications and histone modifications in lung adenocarcinoma (LUAD). RESULTS: Based on the data from public databases, including chromatin property data (ATAC-seq, DNase-seq), methylated RNA immunoprecipitation sequencing (MeRIP-seq), and gene expression data (RNA-seq), a m6A-related differentially expressed gene nerve growth factor inducible (VGF) was identified between LUAD tissues and normal lung tissues. VGF was significantly highly expressed in LUAD tissues and cells, and was associated with a worse prognosis for LUAD, silencing of VGF inhibited the malignant phenotype of LUAD cells by inactivating the PI3K/AKT/mTOR pathway. Through the weighted correlation network analysis (WGCNA) and integration of TCGA-LUAD RNA-seq and m6A methyltransferase METTL3-knockdown RNA-seq data, a significant positive correlation between METTL3 and VGF was observed. By using the MeRIP-qPCR and dual-luciferase reporter assays, we demonstrated that METTL3 knockdown decreased m6A modification level of VGF coding sequences in LUAD cells, the colorimetric m6A quantification assay also showed that METTL3 knockdown significantly decreased global m6A modification level in LUAD cells. Interestingly, we found that METTL3 knockdown also reduced VGF expression by increasing H3K36me3 modification at the VGF promoter. Further research revealed that METTL3 knockdown upregulated the expression of histone methylase SETD2, the major H3K36me3 methyltransferase, by methylating the m6A site in the 3'UTR of SETD2 mRNA in LUAD cells. CONCLUSIONS: Overall, our results reveal that the expression of VGF in LUAD cells is regulated spatio-temporally by METTL3 through both transcriptional (via histone modifications) and post-transcriptional (via m6A modifications) mechanisms. The synergistic effect of these multiple epigenetic mechanisms provides new opportunities for the diagnosis and precision treatment of tumors.

The KdmB-EcoA-RpdA-SntB (KERS) chromatin regulatory complex controls development, secondary metabolism and pathogenicity in Aspergillus flavus.

The filamentous fungus Aspergillus flavus is a plant and human pathogen predominantly found in the soil as spores or sclerotia and is capable of producing various secondary metabolites (SM) such as the carcinogenic mycotoxin aflatoxin. Recently, we have discovered a novel nuclear chromatin binding complex (KERS) that contains the JARID1-type histone demethylase KdmB, a putative cohesion acetyl transferase EcoA, a class I type histone deacetylase RpdA and the PHD ring finger reader protein SntB in the model filamentous fungus Aspergillus nidulans. Here, we show the presence of the KERS complex in A. flavus by immunoprecipitation-coupled mass spectrometry and constructed kdmBΔ and rpdAΔ strains to study their roles in fungal development, SM production and histone post-translational modifications (HPTMs). We found that KdmB and RpdA couple the regulation of SM gene clusters with fungal light-responses and HPTMs. KdmB and RpdA have opposing roles in light-induced asexual conidiation, while both factors are positive regulators of sclerotia development through the nsdC and nsdD pathway. KdmB and RpdA are essential for the productions of aflatoxin (similar to findings for SntB) as well as cyclopiazonic acid, ditryptophenaline and leporin B through controlling the respective SM biosynthetic gene clusters. We further show that both KdmB and RpdA regulate H3K4me3 and H3K9me3 levels, while RpdA also acts on H3K14ac levels in nuclear extracts. Therefore, the chromatin modifiers KdmB and RpdA of the KERS complex are key regulators for fungal development and SM metabolism in A. flavus.

Establishment of an NPK nutrient monitor system in yield-graded cotton petioles under drip irrigation.

BACKGROUND: The determination of nutrient content in the petiole is one of the important methods for achieving cotton fertilization management. The establishment of a monitoring system for the nutrient content of cotton petioles during important growth periods under drip irrigation is of great significance for achieving precise fertilization and environmental protection. METHODS: A total of 100 cotton fields with an annual yield of 4500-7500 kg/ha were selected among the main cotton-growing areas of Northern Xinjiang. The nitrate nitrogen (NO3--N), inorganic phosphorus (PO43--P) and inorganic potassium (K+-K) content and yield of cotton petioles were recorded. Based on a yield of 6000 kg/ha as the dividing line, a two-level and yield-graded monitoring system for NO3--N, PO43--P and K+-K in cotton petioles during important growth periods was established, and predictive yield models for NO3--N, PO43--P and K+-K in petioles during important growth periods were established. RESULTS: The results showed found that the yields of the 100 cotton fields surveyed were normally distributed. Therefore, two yield grades were classified using 6000 kg/ha as a criterion. Under different yield-graded, the NO3--N, PO43--P and K+-K content of petiole at important growth stages was significantly positively correlated with yield. Further, the variation range of NO3--N, PO43--P and K+-K content in petioles could be used as a standard for yield-graded. In addition, a yield prediction model for the NO3--N, PO43--P and K+-K content of petioles was developed. The SSO-BP validation model performed the best (R2 = 0.96, RMSE = 0.06 t/ha, MAE = 0.05 t/ha) in the full bud stage, which was 12.9% higher than the BP validation model. However, the RMSE and MAE were decreased by 86.7% and 88.1%, respectively. CONCLUSION: The establishment of NPK nutrition monitor system of cotton petioles under drip irrigation based on yield-graded provides an important basis for nutrition monitor of cotton petiole under drip irrigation in Xinjiang. It also provides a new method for cotton yield prediction.

Size effect of graphene oxide from quantum dot to nanoflake on the mobility of nanoplastics in seawater-saturated sand.

Marine sedimentary environment serves as an important sink of terrigenous nanoplastics (NP) and graphene oxides (GO). In this study, we discovered that GO of varying sizes exhibited distinct binding modes with 200 nm NP in 35 practical salinity unit (PSU) seawater, resulting in varying impacts on the mobility of NP in porous media. GO-8, with a size of 8±2 nm, firmly adhered to the surface of NP and formed stable primary heterogeneous aggregates, which promoted NP mobility and increased the mass recovery of effluent (Meff) from 24.74% to 31.08%. GO-250 (246±10 nm) partly enveloped NP and only slightly increased the volume of heteroaggregates, which had minimal effect on NP transport. Conversely, GO-850 (855±55 nm) wrapped numerous NP particles to form large secondary heteroaggregates that clung to sand surfaces, providing additional attachment sites for NP, resulting in complete inhibition of NP mobility in porous media (Meff = 0%). In brackish water with 3.5 PSU, all GO-8, GO-250 and GO-850 achieved enhanced mobility of NP, with Meff increasing from 50.35% to 85.62%, 69.45% and 75.41%, respectively. The results indicate that GO size effects on NP mobility are also salinity-dependent.

Effect of different NPK fertilization timing sequences management on soil-petiole system nutrient uptake and fertilizer utilization efficiency of drip irrigation cotton.

In order to elucidate the effects of different nitrogen (N), phosphorus (P), and potassium (K) fertilization timing sequences management on nutrient absorption and utilization in drip irrigation cotton, field experiments were conducted from 2020 to 2021. There are six timing sequences management methods for NPK fertilization (S1-S6: 1/3Time N-1/3Time PK-1/3Time W, 1/3Time PK-1/3Time N-1/3Time W, 1/2Time NPK-1/2Time W, 1/4Time W-1/4Time N-1/4Time PK-1/4Time W, 1/3Time W-1/3Time NPK-1/3Time W), among which S6 is the current management method for field fertilization timing sequences, and S7 is the non N. The results showed that during the main growth stage, S5 accumulated more nitrate nitrogen (NO3--N) and ammonium nitrogen (NH4+-N) content in soil between 20 and 40 cm, and accumulated more available phosphorus content in soil between 5-15 cm and 15-25 cm, S5 reducing N leaching and increasing P mobility. It is recommended to change the timing sequences management method of NPK fertilization for drip irrigation cotton to 1/4Time W-1/4Time PK-1/4Time N-1/4Time W, which is beneficial for plant nutrient absorption and utilization while reducing environmental pollution.