CircEYA3 aggravates intervertebral disc degeneration through the miR-196a-5p/EBF1 axis and NF-κB signaling.

Intervertebral disc degeneration (IDD) is a well-established cause of disability, and extensive evidence has identified the important role played by regulatory noncoding RNAs, specifically circular RNAs (circRNAs) and microRNAs (miRNAs), in the progression of IDD. To elucidate the molecular mechanism underlying IDD, we established a circRNA/miRNA/mRNA network in IDD through standardized analyses of all expression matrices. Our studies confirmed the differential expression of the transcription factors early B-cell factor 1 (EBF1), circEYA3, and miR-196a-5p in the nucleus pulposus (NP) tissues of controls and IDD patients. Cell proliferation, apoptosis, and extracellular mechanisms of degradation in NP cells (NPC) are mediated by circEYA3. MiR-196a-5p is a direct target of circEYA3 and EBF1. Functional analysis showed that miR-196a-5p reversed the effects of circEYA3 and EBF1 on ECM degradation, apoptosis, and proliferation in NPCs. EBF1 regulates the nuclear factor kappa beta (NF-кB) signalling pathway by activating the IKKß promoter region. This study demonstrates that circEYA3 plays an important role in exacerbating the progression of IDD by modulating the NF-κB signalling pathway through regulation of the miR196a-5p/EBF1 axis. Consequently, a novel molecular mechanism underlying IDD development was elucidated, thereby identifying a potential therapeutic target for future exploration.

Contrasting fate and binding behavior of Mn and Cu with dissolved organic matter during in situ remediation using multicomponent capping in malodorous black water.

The common use of peroxides in the remediation of malodorous black water may lead to the activation of heavy metals in sediment when eliminating black and odorous substances. The mechanisms of heavy metal interactions with dissolved organic matter (DOM) in response to in situ capping have not been elucidated, but this information could guide the optimization of capping materials. We developed a capping material consisting of hydrothermally carbonized sediment (HCS), hydrated magnesium carbonate (HMC) and sodium percarbonate (SPC) and used microcosm experiments to investigate the dynamics of Mn and Cu at the sediment-water interface in malodorous black water. The results showed that HCS, HMC and SPC contributed multiple functions of mechanical protection, chemical isolation and oxygen provision to the new caps. HMC promoted the conversion of Mn/Cu into carbonate minerals. The optimal mass proportions were 25 % HCS, 60 % HMC and 15 % SPC based on the mixture design. In situ capping altered the fate and transformation of metals in the sediment-overlying water profile in the short term through Mn immobilization and Cu activation. The complexation of Cu(II) ions was significantly stronger than that of Mn(II) ions. In situ capping had a significant effect on the order of complexation of different fluorescent DOM molecules with Mn(II)/Cu(II) ions: microbial byproducts and fulvic acid-like components were preferentially complexed with Cu(II) ions after capping, while phenolic and humic acid-like components preferentially interacted with Mn(II) ions. Humic-like components bound to Cu were affected the most by capping treatment, whereas protein-like components were relatively weakly affected. Our study provides valuable knowledge on the impact of in situ capping on DOM-metal complexes.

COVID-19 vaccination willingness among people living with HIV in Shijiazhuang, China: a cross-sectional survey.

Objectives: The COVID-19 pandemic imposed an enormous disease and economic burden worldwide. SARS-CoV-2 vaccination is essential to containing the pandemic. People living with HIV (PLWH) may be more vulnerable to severe COVID-19 outcomes; thus, understanding their vaccination willingness and influencing factors is helpful in developing targeted vaccination strategies. Methods: A cross-sectional study was conducted between 15 June and 30 August 2022 in Shijiazhuang, China. Variables included socio-demographic characteristics, health status characteristics, HIV-related characteristics, knowledge, and attitudes toward COVID-19 vaccination and COVID-19 vaccination status. Multivariable logistic regression was used to confirm factors associated with COVID-19 vaccination willingness among PLWH. Results: A total of 1,428 PLWH were included, with a 90.48% willingness to receive the COVID-19 vaccination. PLWH were more unwilling to receive COVID-19 vaccination for those who were female or had a fair/poor health status, had an allergic history and comorbidities, were unconvinced and unsure about the effectiveness of vaccines, were unconvinced and unsure about the safety of vaccines, were convinced and unsure about whether COVID-19 vaccination would affect ART efficacy, or did not know at least a type of domestic COVID-19 vaccine. Approximately 93.00% of PLWH have received at least one dose of the COVID-19 vaccine among PLWH, and 213 PLWH (14.92%) reported at least one adverse reaction within 7 days. Conclusion: In conclusion, our study reported a relatively high willingness to receive the COVID-19 vaccination among PLWH in Shijiazhuang. However, a small number of PLWH still held hesitancy; thus, more tailored policies or guidelines from the government should be performed to enhance the COVID-19 vaccination rate among PLWH.

Discovery of Novel PROTAC Degraders of p300/CBP as Potential Therapeutics for Hepatocellular Carcinoma.

Adenoviral E1A binding protein 300 kDa (p300) and its closely related paralog CREB binding protein (CBP) are promising therapeutic targets for human cancer. Here, we report the first discovery of novel potent small-molecule PROTAC degraders of p300/CBP against hepatocellular carcinoma (HCC), one of the most common solid tumors. Based upon the clinical p300/CBP bromodomain inhibitor CCS1477, a conformational restriction strategy was used to optimize the linker to generate a series of PROTACs, culminating in the identification of QC-182. This compound effectively induces p300/CBP degradation in the SK-HEP-1 HCC cells in a dose-, time-, and ubiquitin-proteasome system-dependent manner. QC-182 significantly downregulates p300/CBP-associated transcriptome in HCC cells, leading to more potent cell growth inhibition compared to the parental inhibitors and the reported degrader dCBP-1. Notably, QC-182 potently depletes p300/CBP proteins in mouse SK-HEP-1 xenograft tumor tissue. QC-182 is a promising lead compound toward the development of p300/CBP-targeted HCC therapy.

Identification of Adulteration of Flaxseed Oil From QINGHAI Area Using GC-MS Profiling of Phytosterol Composition and Chemometrics.

Flaxseed oil is an important source of vegetable oil with a polyunsaturated fatty acid. It is significant to establish a method to quickly identify adulterated flaxseed oil. In the present study, the qualitative and quantitative analysis of phytosterol of flaxseed oil from different varieties and different production areas in the Qinghai area was first performed by gas chromatography-mass spectrometry (GC-MS) and the phytosterol standard profile of flaxseed oil was established. Then, a combination of similarity evaluation and cluster analysis was used to distinguish pure flaxseed oil from flaxseed oil adulterated with concentrations of 10-50% rapeseed oil, peanut oil, sunflower oil, and sesame oil, and discriminant analysis was used to identify the types of adulterated flaxseed oil. The results showed that similarity evaluation combined with cluster analysis can distinguish pure and adulterated flaxseed oil when the concentration of the adulterant was greater than 10%. Discriminant analysis models accurately identified the types of adulterating oil in flaxseed oil when the concentration of rapeseed, peanut, or sunflower oil was greater than 20%, and that of sesame oil was greater than 30%. This study shows that the determination of the phytosterol composition and chemometrics is a valuable tool to evaluate the purity of flaxseed oil.

An improved Back Propagation Neural Network framework and its application in the automatic calibration of Storm Water Management Model for an urban river watershed.

The use of the Storm Water Management Model (SWMM) to simulate flows in urban river watersheds necessitates the proper calibration of the various parameters involved in the process. Back Propagation Neural Network (BPNN) is often used to establish relationship between two sets of multivariate variables, such as parameters and simulation results of SWMM. The aim of this study is to establish an improved BPNN to calibrate SWMM. It was found that when using gauged flow data obtained from the urban river management system as calibration data, only using BPNN was not sufficient. An improved BPNN framework was proposed with integrating Principal Component Analysis (PCA) and Genetic Algorithm (GA) process, abbreviated as PCA-GA-BPNN. It was proved to be effective for calibration. The BPNN combined with GA process made 90 % of the predicted parameters within reasonable range, which was only 8 % using BPNN alone. The PCA process reduced the training time up to 64 %. Using a hydrograph of 196 h, compared with the nondominated sorting genetic algorithm (NSGA), PCA-GA-BPNN training time can be reduced from 18,142 s down to 4.5 s. Nash efficiency coefficients (NSE) of hydrographs fitting was 0.75. Including more rainfall events data in calibration achieved better fitting than including more gauging station data.

Formyl phloroglucinol meroterpenoids from the leaves of Eucalyptus globulus subsp. maidenii and their ATP-citrate lyase inhibitory activities.

Three new formyl phloroglucinol meroterpenoids, eumaidials A-C (1-3), were isolated from the leaves of Eucalyptus globulus subsp. maidenii, along with ten known analogues (4-13). Their chemical structures were determined by various spectral data and electronic circular dichroism calculations. Eumaidial A (1) is the first ß-caryophyllene-based formyl phloroglucinol meroterpenoids from the genus Eucalyptus. Compounds 1-4 and 10 exhibited ATP-citrate lyase inhibitory activities, and compounds 2 and 3 suppressed the hepatocyte lipogenesis.

Design and synthesis of cantharidin derivative DCZ5418 as a TRIP13 inhibitor with anti-multiple myeloma activity in vitro and in vivo.

Natural product cantharidin can inhibit multiple myeloma cell growth in vitro, while serious adverse effects limited its clinical application. Therefore, the structural modification of cantharidin is needed. Herein, inspired by the structural similarity of the aliphatic endocyclic moiety in cantharidin and TRIP13 inhibitor DCZ0415, we designed and synthesized DCZ5418 and its nineteen derivatives. The molecular docking study indicated that DCZ5418 had a similar binding mode to TRIP13 protein as DCZ0415 while with a stronger docking score. Moreover, the bioassay studies of the MM-cells viability inhibition, TRIP13 protein binding affinity and enzyme inhibiting activity showed that DCZ5418 had good anti-MM activity in vitro and definite interaction with TRIP13 protein. The acute toxicity test of DCZ5418 showed less toxicity in vivo than cantharidin. Furthermore, DCZ5418 showed good anti-MM effects in vivo with a lower dose administration than DCZ0415 (15 mg/kg vs 25 mg/kg) on the tumor xenograft models. Thus, we obtained a new TRIP13 inhibitor DCZ5418 with improved safety and good activity in vivo, which provides a new example of lead optimization by using the structural fragments of natural products.

Design, synthesis of auristatins-glucuronide conjugates targeting the ß-glucuronidase in tumor microenvironment.

Auristatins-glucuronide conjugates designed targeting the ß-Glucuronidase in tumor microenvironment were synthesized and evaluated on stabilities, the release of auristatins and the antitumor activities in this study. Conjugates 20 and 21 showed remarkable stabilities in phosphate buffer and bovine serum solution, and excellent selectivity between the in vitro antiproliferative activities against ß-glucuronidase pretreated and untreated cancer cells (IC50 = 5.7 nM âˆ¼ 9.7 nM, IC50 (-Enz) > 1 µM). Furthermore, conjugate 20 showed potent antitumor efficacy in HCT-116 xenograft mouse model without inducing side effects.

Synthesis, evaluation and molecular dynamics study of human toll-like receptor 2/6 specific monoacyl lipopeptides as candidate immunostimulants.

TLR2 agonists typified by the S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-R-cysteinyl-S-serine (Pam2CS) motif have exhibited powerful immunostimulatory activities. Based on simplified monoacyl lipopeptide (Carbamate-linked N-Ac PamCS), we describe interesting SAR investigations where modifications are done to alter the size of substituents on the cysteine amine, introduce ionizable groups to the terminal and insert aromatic substitutions to the aliphatic chain. Our structural modifications have led to a highly specific human TLR2/6 agonist 14a (EC50 = 0.424 nM), which behaves like Pam2CSK4 by inducing NF-κB activation to trigger downstream signaling pathways, such as subsequent phosphorylation of related proteins (p65, p38) and production of key inflammatory cytokines (IL-6, IL-1ß, TNF-α). Importantly, the ability to stimulate enhanced T cell response compared to Carbamate-linked N-Ac PamCS makes compound 14a a further potential candidate immunostimulant.

N6-methyladenosine modification of REG1α facilitates colorectal cancer progression via ß-catenin/MYC/LDHA axis mediated glycolytic reprogramming.

Aerobic glycolysis has been considered as a hallmark of colorectal cancer (CRC). However, the potential functional regulators of glycolysis in CRC remains to be elucidated. In the current study, we found that Regenerating islet-derived protein 1-alpha (REG1α) was significantly increased in both CRC tissues and serum, and positively associated with CRC patients' lymph node metastasis, advanced tumor stage, and unfavorable prognosis. Ectopic expression of REG1α contributed to various tumorigenic properties, including cell proliferation, cell cycle, migration, invasion, and glycolysis. In contrast, REG1α deficiency in CRC cells attenuated malignant properties and glucose metabolism. Mechanically, REG1α promoted CRC proliferation and metastasis via ß-catenin/MYC axis-mediated glycolysis upregulation. Moreover, the malignant behaviors governed by REG1α could be effectively abolished by silencing of Wnt/ß-catenin/MYC axis or glycolysis process using specific inhibitors. Besides, REG1α expression was mediated by METTL3 in an m6A-dependent manner. Overall, our work defines a novel regulatory model of the METTL3/REG1α/ß-catenin/MYC axis in CRC, which indicates that REG1α could function as a novel biomarker and a potential therapeutic target for patients with CRC.

Levels, tissue distribution and isomer stereoselectivity of Dechlorane Plus in humans: A critical review.

Exposure of human tissues to Dechlorane Plus (DP) has raised public concern because of the multiple health threats it may pose to humans. Therefore, it is important to summarize the main findings of previous studies on DP in human tissues and to provide potential guidance for future studies. In this paper, DP levels in different populations and human tissues worldwide since 2009 were systematically reviewed. DP levels in human tissues of workers in e-waste dismantling sites in Guangdong Province, China (median 190 ng·g-1 lw in serum) and DP manufacturing plants in Jiangsu Province, China (mean 857 ng·g-1 lw in whole-blood) are the highest reported worldwide. DP levels in tissues of the general population in recent studies are close to those of residents near e-waste dismantling sites, which should be of concern. DP levels in different human tissues were found to be positively correlated with a pattern of blood > breast milk > adipose tissue. The distribution of DP in different human tissues is mainly lipid-driven and may also be influenced by the interaction of DP with proteins such as human serum albumin. Most of the past studies determined the isomer stereoselectivity of DP in human tissues only by comparing the composition of DP in commercial DP products and human tissues, which lacks evidence of mechanism. Recently, a significantly different affinity of DP isomers for proteins was found, which seems to confirm the isomer selectivity of DP in human tissues. We simulated the binding of DP to human serum albumin and DP to thyroid hormone receptor ß by molecular docking and found differences in the binding behavior of syn-DP and anti-DP to the selected proteins. Molecular docking seems to be a feasible approach for future studies to predict and reveal the mechanisms of DP behavior and health effects in human tissues.

5-HMF attenuates inflammation and demyelination in experimental autoimmune encephalomyelitis mice by inhibiting the MIF-CD74 interaction.

The neuroprotective role of 5-hydroxymethyl-2-furfural (5-HMF) has been demonstrated in a variety of neurological diseases. The aim of this study is to investigate the effect of 5-HMF on multiple sclerosis (MS). IFN-γ-stimulated murine microglia (BV2 cells) are considered a cell model of MS. With 5-HMF treatment, microglial M1/2 polarization and cytokine levels are detected. The interaction of 5-HMF with migration inhibitory factor (MIF) is predicted using online databases. The experimental autoimmune encephalomyelitis (EAE) mouse model is established, followed by a 5-HMF injection. The results show that 5-HMF facilitates IFN-γ-stimulated microglial M2 polarization and attenuates the inflammatory response. According to the network pharmacology and molecular docking results, 5-HMF has a binding site for MIF. Further results show that blocking MIF activity or silencing CD74 enhances microglial M2 polarization, reduces inflammatory activity, and prevents ERK1/2 phosphorylation. 5-HMF inhibits the MIF-CD74 interaction by binding to MIF, thereby inhibiting microglial M1 polarization and enhancing the anti-inflammatory response. 5-HMF ameliorates EAE, inflammation, and demyelination in vivo. In conclusion, our research indicates that 5-HMF promotes microglial M2 polarization by inhibiting the MIF-CD74 interaction, thereby attenuating inflammation and demyelination in EAE mice.

A multiscale analysis of the spatially heterogeneous relationships between non-point source pollution-related processes and their main drivers in Chaohu Lake watershed, China.

A better understanding of the relationships between non-point source (NPS) pollution-related processes and their drivers will help to develop scientific watershed management measures. Although various studies have explored the drivers' impact on NPS pollution-related processes, quantitative knowledge of the properties within these relationships is still needed. This study uses the Integrated Valuation of Ecosystem Services and Trade-offs (InVEST) model to produce three related processes of NPS pollution, quick flow (QF), nitrogen export (NE), and sediment export (SE), in the upstream watershed of Chaohu Lake, China. The spatial distributions of QF, NE, and SE and their responses to multiple natural-socioeconomic drivers at nine spatial scales (1 km2, 10 km2, 20 km2, 30 km2, 50 km2, 75 km2, 100 km2, 200 km2, and town) were compared. The results showed that the spatial scale has little impact on the spatial distributions of NPS pollution-related processes. Across the nine scales, the socioeconomic drivers related to agricultural activities, area proportions of cultivated land (cultivated) and paddy field (paddy), have dominant impacts on NE, while the topographical drivers, the connectivity index (IC) and slope, have dominant impacts on both SE and QF. The magnitudes of single and paired natural-socioeconomic drivers' impacts on NPS pollution-related processes increase logarithmically or linearly with increasing spatial scale, but they tend to reach a stable threshold at a certain coarse scale. Our results emphasized the necessity and importance of embracing spatial scale effects in watershed water environmental management.

An in-depth investigation of the influence of sample size on PCA-MLR, PMF, and FA-NNC source apportionment results.

The mechanism by which parameters influence the source apportionment results of receptor models is not well understood. Three mature receptor models, namely, principal component analysis-multiple linear regression (PCA-MLR), positive matrix factorization (PMF) and factor analysis with nonnegative constraints (FA-NNC), were comparatively employed for source apportionment of 16 polycyclic aromatic hydrocarbons in 30 street dust samples. The results indicated that the FA-NNC and PMF models produced results with a higher degree of similarity than the results obtained with the PCA-MLR model. Moreover, when the sample size was gradually decreased, similar source profiles were extracted that were consistent with results obtained from all samples. However, the overall contribution rates were not as stable as the source profiles. The PCA-MLR results remained the most stable in both aspects. FA-NNC and PMF performed better in regards to the stability of contribution rates and source profiles, respectively. Improvements in the goodness of fit of overall and individual pollutants were always accompanied by a decrease in the relevance among the variables, indicating that while the model simulation effect was improved, the credibility of the results decreased. Thus, finding an appropriate number of sample size is more appropriate than simply involving too many samples in source apportionment models.

Identification and verification of pivotal genes promoting the progression of atherosclerosis based on WGCNA.

As the main pathological basis for the development of cardiovascular and cerebrovascular diseases, atherosclerosis (AS) seriously affects human health. The key targets of biological information analysis of AS can help exploit therapeutic targets. The expression data of early and progressive atherosclerotic tissues were downloaded from the Gene Expression Omnibus (GEO) database. Based on GSE28829 and GSE120521, 74 key genes were obtained through differential expression analysis and weighted correlation network analysis (WGCNA) analysis, which were mainly enriched in the regulating of inflammatory response, chemokine signalling pathway, apoptosis, lipid and AS, Toll-like receptor signalling pathway and so on according to the results of the enrichment analysis. Cytoscape software was applied to screen four pivotal genes (TYROBP, ITGB2, ITGAM and TLR2) based on PPI. The results of the correlation analysis showed that the expression level of pivotal genes was positively related to macrophages M0, and was negatively related to T cells follicular helper. In addition, the expression of ITGB2 was positively related to Tregs. In this study, bioinformatics was applied to screen pivotal genes affecting the progress of AS, which were significantly related to immune-related biological functions and signal pathways of atherosclerotic tissues and the infiltration level of immune cells. Therefore, pivotal genes were expected to become therapeutic targets for AS.

Optimization of potent, selective and orally bioavailable biphenyl scaffold as FABP4 inhibitors for anti-inflammation.

Fatty-acid binding protein 4 (FABP4) is an essential driver for the progression of metabolic-related inflammatory diseases including obesity, diabetes, atherosclerosis, and various lipid metabolism-related tumors. However, FABP4 inhibitors are not yet available for clinical use, which may be associated with their poor selectivity of FABP3, unsatisfactory efficacy and physicochemical properties. Herein, we reported a systematic optimization of a class of biphenyl scaffold molecules as potent FABP4 inhibitors. Further in vitro and in vivo pharmacokinetic studies identified a selective and orally bioavailable compound 10g, with Ki of 0.51 µM against FABP4, Ki of 33.01 µM against FABP3 and bioavailability F% value of 89.4%. In vivo anti-inflammatory efficacy and multi-organ protection study in LPS-induced inflammatory mice model highlighted the potential of compound 10g as a therapeutic candidate in inflammation-related diseases.

Performance of 2D-shear wave elastography in autoimmune hepatitis-primary biliary cholangitis overlap syndrome.

PURPOSE: To evaluate the diagnostic values of liver stiffness (LS) measured by 2D-SWE, fibrosis index based on the four factors (FIB-4), aspartate aminotransferase to platelet ratio index (APRI), and GGT to PLT ratio (GPR) for assessing liver fibrosis and high-risk esophageal varices (EVs) in patients with autoimmune hepatitis-primary biliary cholangitis (AIH-PBC) overlap syndrome. METHODS: Data of 141 patients were retrospectively collected. Liver fibrosis was staged according to the Scheuer scoring system. The Spearman correlation coefficient was used for correlation analysis. Receiver operating characteristic (ROC) curves were plotted to evaluate the diagnostic performance. RESULTS: LS and FIB-4 were positively correlated with the fibrosis stage (r = 0.555 and 0.198, respectively). LS had significantly higher areas under the ROC curves (AUROCs) values than FIB-4 for predicting advanced fibrosis (0.818 vs. 0.567, P < 0.001), cirrhosis (0.879 vs. 0.637, P < 0.001), whereas LS and FIB-4 similarly predicted significant fibrosis (0.748 vs. 0.638, P = 0.071) and high-risk EVs (0.731 vs. 0.659, P = 0.303). The optimal cut-off values of 2D-SWE for detecting significant fibrosis, advanced fibrosis, cirrhosis, and high-risk EVs were 8.7 kPa, 12.8 kPa, 14.0 kPa, and 11.0 kPa, respectively. LS values were influenced by fibrosis stage, serum GGT, albumin, and total bilirubin levels. The overall concordance rate of the liver stiffness vs. Scheuer stages was 49.65%. CONCLUSIONS: 2D-SWE shows significantly greater diagnostic accuracy than serum fibrosis indexes for diagnosing advanced fibrosis and cirrhosis in patients with AIH-PBC overlap syndrome.

Discovery of Small-Molecule Autophagy Inhibitors by Disrupting the Protein-Protein Interactions Involving Autophagy-Related 5.

One possible strategy for modulating autophagy is to disrupt the critical protein-protein interactions (PPIs) formed during this process. Our attention is on the autophagy-related 12 (ATG12)-autophagy-related 5 (ATG5)-autophagy-related 16-like 1 (ATG16L1) heterotrimer complex, which is responsible for ATG8 translocation from ATG3 to phosphatidylethanolamine. In this work, we discovered a compound with an (E)-3-(2-furanylmethylene)-2-pyrrolidinone core moiety (T1742) that blocked the ATG5-ATG16L1 and ATG5-TECAIR interactions in the in vitro binding assay (IC50 = 1-2 µM) and also exhibited autophagy inhibition in cellular assays. The possible binding mode of T1742 to ATG5 was predicted through molecular modeling, and a batch of derivatives sharing essentially the same core moiety were synthesized and tested. The outcomes of the in vitro binding assay and the flow cytometry assay of those newly synthesized compounds were generally consistent. This work has validated our central hypothesis that small-molecule inhibitors of the PPIs involving ATG5 can tune down autophagy effectively, and their pharmaceutical potential may be further explored.

An improved nonnegative matrix factorization with the imputation method model for pollution source apportionment during rainstorm events.

Data scarcity caused by extreme conditions during storms adds difficulties in performing pollution source apportionment. This study integrated nonnegative matrix factorization with the imputation method (NMF-IM) to fill in missing data (NAs) and conduct source apportionment. A total of 367 river samples and 35 runoff samples were taken from the Banqiao and Nanfei River basins located in Hefei, China, during four rainfall events from June to August 2020. Sixteen indicators were quantified and used for source diagnostics using NMF-IM. The results showed that total phosphorus (TP) had higher concentrations and more violent fluctuations than total nitrogen (TN) in river samples taken from rain. NMF-IM was shown to recover the value distribution of NAs approximately. The source profiles and contribution rates calculated by NMF-IM with NAs were close to the original results calculated by NMF without NAs, with root mean square error of less than 2.3% and differences less than 9.5%. Multiple forms of nitrogen and phosphorus indicators benefit reaching reasonable source diagnostics results. At least four indicators were needed to reach the same contribution rates as 16 indicator diagnostics. The two good indicator combination groups are nitrate (NO3-N), nitrite (NO2-N), ammonia nitrogen (NH3-N), and total suspended solids (TSS) and NO3-N, NO2-N, phosphorus (PO4-P), and TSS. The pollution source contributions changed with the Antecedent dry period (ADPs) of rain events. Treated tailwater and untreated sewage were major sources, contributing more than 80% of the total pollution of the rainstorm events with short ADPs. Dust wash became the dominant contributor after 60 min and contributed 36% of the total pollution of rainstorm events with long ADPs. The average source contribution rates for rainfall events in the Banqiao River were treated tailwater (41%) > untreated sewage (27%) > dust wash (19%) > other sources (16%). The pollution source diagnostics results were verified to be reasonable by simulation using tested run-off data and literature results.