Precise Spectral Overlap-Based Donor-Acceptor Pair for a Sensitive Traffic Light-Typed Bimodal Multiplexed Lateral Flow Immunoassay.

Bimodal-type multiplexed immunoassays with complementary mode-based correlation analysis are gaining increasing attention for enhancing the practicability of the lateral flow immunoassay (LFIA). Nonetheless, the restriction in visually indistinguishable multitargets induced by a single fluorescent color and difficulty in single acceptor ineffectual fluorescence quenching due to the various spectra of multiple different donors impede the further execution of colorimetric-fluorescence bimodal-type multiplexed LFIAs. Herein, the precise spectral overlap-based donor-acceptor pair construction strategy is proposed by regulating the size of the nanocore, coating it with an appropriate nanoshell, and selecting a suitable fluorescence donor with distinct colors. By in situ coating Prussian blue nanoparticles (PBNPs) on AuNPs with a tunable size and absorption spectrum, the resultant APNPs demonstrate efficient fluorescence quenching ability, higher colloidal stability, remarkable colorimetric intensity, and an enhanced antibody coupling efficiency, all of which facilitate highly sensitive bimodal-type LFIA analysis. Following integration with competitive-type immunoreaction, this precise spectral overlap-supported spatial separation traffic light-typed colorimetric-fluorescence dual-response assay (coined as the STCFD assay) with the limits of detection of 0.013 and 0.152 ng mL-1 for ractopamine and clenbuterol, respectively, was proposed. This work illustrates the superiority of the rational design of a precise spectral overlap-based donor-acceptor pair, hinting at the enormous potential of the STCFD assay in the point-of-care field.

Separable Microneedle for Integrated Hyperglycemia Sensing and Photothermal Responsive Metformin Release.

Microdevices that offer hyperglycemia monitoring and controllable drug delivery are urgently needed for daily diabetes management. Herein, a theranostic separable double-layer microneedle (DLMN) patch consisting of a swellable GelMA supporting base layer for glycemia sensing and a phase-change material (PCM) arrowhead layer for hyperglycemia regulation has been fabricated. The Cu-TCPP(Fe)/glucose oxidase composite and 3,3',5,5'-tetramethylbenzidine coembedded in the supporting base layer permit a visible color shift at the base surface in the presence of glucose via a cascade reaction, allowing for the in situ detection of glucose in interstitial fluid. The PCM arrowhead layer is encapsulated with water monodispersity melanin nanoparticles from Sepia officinalis and metformin that is imparted with a near-infrared ray photothermal response feature, which is beneficial to the controllable release of metformin for suppression of hyperglycemia. By applying the DLMN patch to the streptozotocin-induced type 2 diabetic Sprague-Dawley rat model, the results demonstrated that it can effectively extract dermal interstitial fluid, read out glucose levels, and regulate hyperglycemia. This DLMN-integrated portable colorimetric sensor and self-regulated glucose level hold great promise for daily diabetes management.

Engineered Collaborative Size Regulation and Shape Engineering of Tremella-Like Au-MnOx for Highly Sensitive Bimodal-Type Lateral Flow Immunoassays.

Engineered collaborative size regulation and shape engineering of multi-functional nanomaterials (NPs) offer extraordinary opportunities for improving the analysis performance. It is anticipated to address the difficulty in distinguishing color changes caused by subtle variations in target concentrations, thereby facilitating the highly sensitive analysis of lateral flow immunoassays (LFIAs). Herein, tremella-like gold-manganese oxide (Au-MnOx ) nanoparticles with precise MnCl2 regulation are synthesized as immuno signal tracers via a facile one-step redox reaction in alkaline condition at ambient temperature. Avail of the tunable elemental composition and anisotropy in morphology, black-colored tremella-like Au-MnOx exhibits superb colorimetric signal brightness, enhanced antibody coupling efficiency, marvelous photothermal performance, and unrestricted immunological recognition affinity, all of which facilitate highly sensitive multi-signal transduction patterns. In conjunction with the handheld thermal reader device, a bimodal-type LFIA that combines size-regulation- and shape-engineering-mediated colorimetric-photothermal dual-response assay (coined as the SSCPD assay) with a limit of detection of 0.012 ng mL-1 for ractopamine (RAC) monitoring is achieved by integrating Au-MnOx with the competitive-type immunoreaction. This work illustrates the effectiveness of this strategy for establishing high-performance sensing, and the SSCPD assay may be extended to a wide spectrum of future point-of-care (POC) diagnostic applications.

Isolation of Pseudomonas oleovorans Carrying Multidrug Resistance Proteins MdtA and MdtB from Wastewater.

The pollution of industrial wastewater has become a global issue in terms of economic development and ecological protection. Pseudomonas oleovorans has been studied as a bacterium involved in the treatment of petroleum pollutants. Our study aimed to investigate the physicochemical properties and drug resistance of Pseudomonas oleovorans isolated from industrial wastewater with a high concentration of sulfate compounds. Firstly, Pseudomonas oleovorans was isolated and then identified using matrix-assisted flight mass spectrometry and 16S rDNA sequencing. Then, biochemical and antibiotic resistance analyses were performed on the Pseudomonas oleovorans, and a microbial high-throughput growth detector was used to assess the growth of the strain. Finally, PCR and proteomics analyses were conducted to determine drug-resistance-related genes/proteins. Based on the results of the spectrum diagram and sequencing, the isolated bacteria were identified as Pseudomonas oleovorans and were positive to reactions of ADH, MTE, CIT, MLT, ONPG, and ACE. Pseudomonas oleovorans was sensitive to most of the tested antibiotics, and its resistance to SXT and CHL and MIN and TIM was intermediate. The growth experiment showed that Pseudomonas oleovorans had a good growth rate in nutrient broth. Additionally, gyrB was the resistance gene, and mdtA2, mdtA3, mdtB2, mdaB, and emrK1 were the proteins that were closely associated with the drug resistance of Pseudomonas oleovorans. Our results show the biochemical properties of Pseudomonas oleovorans from industrial wastewater with a high concentration of sulfate compounds and provide a new perspective for Pseudomonas oleovorans to participate in biological removal of chemical pollutants in industrial wastewater.

Metal-Organic Framework-Loaded Engineering DNAzyme for the Self-Powered Amplified Detection of MicroRNA.

DNAzyme shows great promise in designing a highly sensitive and specific sensing platform; however, the low cellular uptake efficiency, instability, and especially the insufficient cofactor supply inhibit the intracellular molecule sensor applications. Herein, we demonstrate a novel type of DNAzyme-based self-driven intracellular sensor for microRNA (miRNA) detection in living cells. The sensor consists of a metal-organic framework [zeolite imidazole framework (ZIF-8)] core loaded with a shell consisting of a rationally designed DNAzyme, where the substrate strand is modified with FAM and BHQ-1 nearby both the sides of the restriction site, respectively, while the enzyme strand consists of two separate strands with a complementary fragment to the substrate strand and the targeting miRNA, respectively. The ZIF-8 nanoparticles enable the efficient delivery of DNAzyme into the cell and protect the DNAzyme from degradation. The pH-responsive ZIF-8 degradation is accompanied with the release of the DNAzyme and Zn2+ cofactors, and the intracellular target miRNAs recognize and activate the DNAzyme driven by the Zn2+ cofactors to cleave the substrate strand, resulting in the release of the FAM-labeled shorter product strand and increased fluorescence for miRNA detection. The self-driven approach can be generally applied to various miRNAs' detection through DNAzyme engineering.

Solvent-controlled two-step one-pot syntheses of α-X (X = Br or Cl) enamino ketones/esters and 3-(2,5-dioxopyrrolidin-1-yl)acrylate by using terminal carbonyl alkynes.

A new two-step one-pot aminobromination/chlorination of carbonyl alkynes has been achieved via a Michael addition of aliphatic secondary amines and subsequent ß-bromination/chlorination of the obtained enamines to afford various α-X (X = Br or Cl) enamino ketones/esters in moderate to good yields. A solvent-controllable protocol has been developed to produce versatile 3-(2,5-dioxopyrrolidin-1-yl)acrylates in moderate yields by using toluene as the solvent and chain alkyl propiolates as alkynyl substrates.

Prevalence and molecular identification of Balantioides coli isolates from pet guinea pigs in Central China.

Balantioides coli is the only known zoonotic ciliate that can infect humans and is usually acquired from swine. It has, however, been reported in other mammals, including guinea pigs, where infection prevalence and molecular characterization are relatively unknown. In the present study, 32 guinea pigs from two different pet markets in Luoyang city of the Henan province in China were evaluated for ciliate-like trophozoites or cysts by direct fecal smear microscopy. Positive samples were further characterized using 18S rDNA and ITS1-5.8S rDNA-ITS2 sequence analysis. Microscopy indicated that ciliate-like cysts were observed in the fecal samples of several guinea pigs, were spherical in shape, and exhibited sizes of 40-65 µm in diameter. The average cyst-positive prevalence in guinea pigs was 62.5%. Sequence analysis indicated that the guinea pig-derived ciliate isolates belonged to B. coli and included two genetic variants (A and B), of which genetic variant A was more dominant among the guinea pig samples. To the best of our knowledge, the present study is the first molecular identification of B. coli in guinea pigs and provides some important information for investigating the molecular epidemiology of B. coli.

N1-Selective alkenylation of 1-sulfonyl-1,2,3-triazoles with alkynes via gold catalysis.

A N1-selective alkenylation of 1-sulfonyl-1,2,3-triazoles with alkynes via gold catalysis is reported. N1-Vinyl substituted 1,2,3-triazoles were selectively prepared in up to 92% yield through the sulfonyl group of 1,2,3-triazole derivatives transformed to alkenyl groups in a "one-pot two steps" manner. This method provided a new method for the synthesis of potentially biological-active vinyl-triazole building blocks.

Regioselective 1,2-addition of allenamides with N-haloimides: synthesis of 2-halo allylic aminal derivatives.

A strategy for the synthesis of 2-halo allylic aminal derivatives through regioselective 1,2-addition of allenamides with N-haloimides is presented. This reaction was conducted under very mild conditions and gave up to 99% yield. Moreover, the obtained halides allow functional group diversification by palladium-catalyzed coupling reactions, which could act as potential intermediates for the synthesis of valuable compounds.

[Recent advances in pyridoxal phosphate-dependent enzymes and their applications].

Pyridoxal phosphate (PLP), the active form of vitamin B6, is an important coenzyme in various enzyme-catalyzed reactions. PLP-dependent enzymes can catalyze a variety of chemical reactions, such as racemization, decarboxylation, ß-addition, ß-elimination, retro-aldol cleavage, transamination, and α-elimination. They are biologically synthesized a powerful tool for a variety of natural amino acids, non-natural amino acids and their related compounds. This article details the structural features and catalytic mechanisms of typical PLP-dependent enzymes such as ω-transaminase, lysine decarboxylase, threonine aldolase, and L-tyrosine phenol-lyase, and reviews the research progress in molecular modification and industrial applications of these enzymes. Finally, this article provides an outlook on the future development of PLP-dependent enzymes, including in vivo regeneration system and industrial applications of PLP cofactors, and discusses the tremendous potential of these enzymes in biocatalytic applications.

Identification of molecular subtypes, prognostic status and immunotherapy response in cervical cancer based on angiogenic signature genes.

Background: Cervical cancer, as one of the most common malignancies in women, is closely related to the mechanism of angiogenesis, which needs further exploration. Methods: The squamous cell carcinoma of the cervix and cervical adenocarcinoma (CESC) data from The Cancer Genome Atlas (TCGA) database. CESC subtypes based on 48 angiogenesis-related genes were identified using consistent cluster analysis, and the limma package were adopted to screen the differentially expressed genes (DEGs) associated with prognosis. Further compress the DEGs through univariate and Least Absolute Shrinkage and Selection Operator (LASSO) COX analysis to identify the key genes. Calculate immune scores using the GSVA package and predict immunotherapy response with TIDE. For in vitro analysis, the expressions of these key genes were additionally tested via reverse-transcription quantitative PCR, and the migration and invasion of Hela cells were determined in scratch and transwell assays, respectively. Results: 3 CESC subtypes were identified, with the best survival advantage in the C2 subtype and the worst in C1 subtype. A risk model was established utilizing seven key genes (MMP3, DLL4, CAP2, PDIA6, TCN2, PAPSS2, and VCAM1), showcases an Area Under the Curve (AUC) exceeding 0.7, underlining its robust performance. The risk score model showed a trend of poorer survival for patients in the high-risk score group and good agreement across different datasets. A nomogram was constructed, and calibration curves indicated robust predictive performance. Immunological analysis revealed heightened sensitivity to immunotherapy in the low-risk group. Besides, the elevated expressions of all 7 genes were seen in Hela cells, and the specific target-mediated DLL4 knockdown diminished the migration and invasion of Hela cells in vitro. Conclusion: This research provides fresh insights and a valuable tool to guide therapeutic decision-making for CESC.

Simultaneous Detection of Clenbuterol and Higenamine in Urine Samples Using Interference-Free SERS Tags Combined with Magnetic Separation.

Sports doping remains a significant challenge in competitive sports. Given that urine analysis is the standard for detecting doping, developing rapid, sensitive, accurate, and high-throughput methods for stimulant detection in urine is crucial. Surface-enhanced Raman scattering (SERS) tag-based immunoassays have emerged as powerful analytical tools known for their high sensitivity and specificity, holding particular promise for stimulant detection in urine samples. However, both the Raman signals of typical SERS tags and sample matrices are within the Raman fingerprint region (<1800 cm-1), which could lead to spectrum overlap, potentially reducing detection accuracy and sensitivity. By recognizing this, we designed a competitive immunoassay that integrates two types of zero-background SERS tags and magnetic separation. These innovative SERS tags exhibit distinctive Raman peaks within the Raman-silent region (1800-2800 cm-1), effectively mitigating potential spectrum overlap with background sample signals. Moreover, magnetic separation not only enhances operational simplicity but also improves the system's anti-interference capability. Using clenbuterol (CL) and higenamine (HM) as model targets, the SERS-based competitive immunoassay demonstrated sensitive detection of individual CL or HM standards, with limits of detection (LODs) of 0.87 and 0.71 pg/mL, respectively. In multiplex mode, CL and HM can be simultaneously detected with LODs of 1.0 and 0.81 pg/mL, respectively. Furthermore, the recovery rates in urine samples ranged from 83 to 116% (relative standard deviation, RSD ≤ 6.4%) for CL and from 82 to 103% (RSD ≤ 5.1%) for HM, further confirming the reliability of the SERS-based immunoassay for practical applications.

Rational design of PspAlgL to improve its thermostability and anti-biofilm activity against Pseudomonas aeruginosa.

Pseudomonas aeruginosa biofilm enhances tolerance to antimicrobials and immune system defenses. Alginate is an important component of biofilm and a virulence factor of P. aeruginosa. The degradation of alginate by alginate lyases has come to serve as an adjunctive therapeutic strategy against P. aeruginosa biofilm, but poor stability of the enzyme limited this application. Thus, PspAlgL, an alginate lyase, can degrade acetylated alginate but has poor thermostability. The 3D structure of PspAlgL was predicted, and the thermostability of PspAlgL was rationally designed by GRAPE strategy, resulting in two variants with better stability. These variants, PspAlgLS270F/E311P and PspAlgLG291S/E311P, effectively degraded the alginate in biofilm. In addition, compared with PspAlgL, these variants were more efficient in inhibiting biofilm formation and degrading the established biofilm of P. aeruginosa PAO1, and they were also able to destroy the biofilm attached to catheters and to increase the sensitivity of P. aeruginosa to the antibiotic amikacin. This study provides one potential anti-biofilm agent for P. aeruginosa infection.

Rubber additives and relevant oxidation products in groundwater in a central China region: Levels, influencing factors and exposure.

This study investigated the presence of rubber additives and relevant oxidation products (RAROPs) in groundwater in central China's aboveground river region. Seven RAROPs were detected, and their levels in shallow groundwater showed a mild decreasing trend from the area near the Yellow River (Avg: 8.49 ng L-1) to the area on the far bank of the Yellow River (Avg: 5.01 ng L-1). In contrast, deep groundwater's RAROPs contents showed a dramatic decrease to only 0.26 ng L-1. The dominant contaminant was found to be N-(1, 3-dimethylbutyl) -N'-phenyl -p-phenylenediamine (6PPD). The vicinity of the garages and car parks was often characterized as contamination hotspots. Correlation analyses further indicated that aquaculture was likely to be a potential pathway for shallow groundwater contaminant inputs. The amount of RAROPs intake by humans through groundwater is nearly 30 times different due to the imbalanced development between urban and rural areas. Children were the most vulnerable to RAROPs. Therefore, human activities (transportation, waste tire storage, water resource allocation and utilization patterns, diversion of Yellow River water to aquaculture ponds) may exacerbate RAROPs pollution in groundwater by leaching contaminants through the surface soil. These results are important for developing appropriate utilization and protection strategies for groundwater resources in developing countries.

Identification and Validation of IFI44 as a Novel Biomarker for Primary Sjögren's Syndrome.

Background: Primary Sjögren's syndrome (pSS) is an autoimmune condition marked by lymphocyte infiltration in the exocrine glands. Our study aimed to identify a novel biomarker for pSS to improve its diagnosis and treatment. Methods: The gene expression profiles of pSS were obtained from the Gene Expression Omnibus (GEO) database. The specific differentially expressed genes (DEGs) were screened by the Least Absolute Shrinkage and Selection Operator (LASSO), Random Forest (RF), and Recursive Feature Elimination with Support Vector Machines (SVM-RFE). A biomarker was picked out based on correlation and diagnostic performance, the connection between the biomarker and clinical traits and immune infiltrating cells was explored, and the biomarker's protein expression level in the serum of pSS patients was detected by enzyme-linked immunosorbent assay (ELISA). The competitive endogenous RNA (ceRNA) network regulated by the biomarker was predicted to verify the reliability of the biomarker in diagnosing pSS. Results: IFI44, XAF1, GBP1, EIF2AK2, IFI27, and IFI6 showed prominent diagnostic ability, with the high accuracy (AUC = 0.859) and significance (R ≥ 0.8) of IFI44 within the training dataset. IFI44 strongly exhibited a negative correlation with resting NK cells, macrophages M0, and eosinophils, and a positive correlation with activated dendritic cells, naive B cells, and activated CD4 memory T cells. Furthermore, IFI44 was significantly positively correlated with clinical traits such as IgG, SSA, SSB, ANA, and ESSDAI, with its protein expression level in the serum of pSS patients being notably elevated compared to controls (p < 0.001). Finally, the ceRNA regulatory network showed that hsa-miR-944, hsa-miR-9-5p, hsa-miR-126-5p, and hsa-miR-335-3p were significantly targeted IFI44, suggesting that IFI44 may serve as a dependable biomarker for pSS. Conclusion: In this study, we dug out IFI44 as a biomarker for pSS, systematically studied the potential regulatory mechanism of IFI44, and verified its reliability as a biomarker for pSS.

Adequately increasing the wheat B-starch ratio can improve the structure-properties of dough during freeze-thaw cycles: Mechanisms and conformational relations.

To reveal the influence of wheat starch particle size distribution on frozen dough quality, this study reconstituted A/B starch according to 100:0, 75:25, 50:50, 25:75 and 0:100 and prepared reconstituted dough by compounding with gluten proteins. Further, the freeze-thaw cycle of 1, 3, and 9 times for reconstituted dough was performed to investigate its ratio-regulatory role of A- and B-starch. The results showed that the freeze-thaw cycle induced gluten network breakage and starch granule exposure in doughs mainly by disrupting disulfide and hydrogen bonds between gluten protein molecules and upsetting their secondary structures, leading to a reduction in GMP and polymer protein content and an increase in freezing water content. Moreover, a moderate increase (25-50 %) in the B-starch proportion can minimize gluten protein deterioration by freeze-thaw cycles. However, excessive B-starch amounts (75-100 %) can also adversely affect gluten structure. The prepared dumpling wrappers under the 50A-50B ratio showed optimal steaming loss rate, hardness, and chewiness during the freeze-thaw cycle. Correlation analysis indicated that the B-starch ratio and its filling pattern improved dough freeze-thaw deterioration primarily by affecting dough-free sulfhydryl content, protein molecular weight distribution, secondary structure, and ΔH. The results may provide insights and guidelines for product development and storage for frozen pasta.

Within-Session Reliability of fNIRS in Robot-Assisted Upper-Limb Training.

Functional near-infrared spectroscopy (fNIRS) seems opportune for neurofeedback in robot-assisted rehabilitation training due to its noninvasive, less physical restriction, and no electromagnetic disturbance. Previous research has proved the cross-session reliability of fNIRS responses to non-motor tasks (e.g., visual stimuli) and fine-motor tasks (e.g., finger tapping). However, it is still unknown whether fNIRS responses remain reliable 1) in gross-motor tasks, 2) within a training session, and 3) for different training parameters. Hence, this study aimed to investigate the within-session reliability of fNIRS responses to gross-motor tasks for different training parameters. Ten healthy participants were recruited to conduct right elbow extension-flexion in three robot-assisted modes. The Passive mode was fully motor-actuated, while Active1 and Active2 modes involved active engagement with different resistance levels. FNIRS data of three identical runs were used to assess the within-session reliability in terms of the map- ( R2 ) and cluster-wise ( Roverlap ) spatial reproducibility and the intraclass correlation (ICC) of temporal features. The results revealed good spatial reliability ( R2 up to 0.69, Roverlap up to 0.68) at the subject level. Besides, the within-session temporal reliabilities of Slope, Max/Min, and Mean were between good and excellent ( ICC < 0.86). We also found that the within-session reliability was positively correlated with the intensity of the training mode, except for the temporal reliability of HbO in Active2 mode. Overall, our results demonstrated good within-session reliability of fNIRS responses, suggesting fNIRS as reliable neurofeedback for constructing closed-loop robot-assisted rehabilitation systems.

Vegetable extracts: Effective inhibitors of heterocyclic aromatic amines and advanced glycation end products in roasted Mackerel.

The formation of hazardous substances, heterocyclic aromatic amines (HAAs) and advanced glycation end products (AGEs), in roasted mackerel with different cooking temperatures (180, 210, 240 °C) and vegetable extracts (celery, carrot and yam extracts) in a preheated oven was investigated. The results indicated that the introduction of vegetable extracts had inhibitory effects on HAAs and AGEs during thermal processing, especially celery extracts. Benefiting from the addition of vegetable extracts, the roasted mackerel keep high quality against lipid/protein oxidation, avoids nutrition loss of polyunsaturated fatty acids, and flavor is promoted. We also examined the variation of key precursors, including creatine, creatinine, reducing sugars, amino acids and attempted to explain the molecular pathway of inhibition of the formation of the hazardous substances by vegetable extracts. The results provide theoretical support to develop technologies for inhibiting hazardous substances formation during fish processing, which is important for food manufacturers and consumers for producing healthier meat products.

RNA-Cleaving DNAzyme-Based Amplification Strategies for Biosensing and Therapy.

Since their first discovery in 1994, DNAzymes have been extensively applied in biosensing and therapy that act as recognition elements and signal generators with the outstanding properties of good stability, simple synthesis, and high sensitivity. One subset, RNA-cleaving DNAzymes, is widely employed for diverse applications, including as reporters capable of transmitting detectable signals. In this review, the recent advances of RNA-cleaving DNAzyme-based amplification strategies in scaled-up biosensing are focused, the application in diagnosis and disease treatment are also discussed. Two major types of RNA-cleaving DNAzyme-based amplification strategies are highlighted, namely direct response amplification strategies and combinational response amplification strategies. The direct response amplification strategies refer to those based on novel designed single-stranded DNAzyme, and the combinational response amplification strategies mainly include two-part assembled DNAzyme, cascade reactions, CHA/HCR/RCA, DNA walker, CRISPR-Cas12a and aptamer. Finally, the current status of DNAzymes, the challenges, and the prospects of DNAzyme-based biosensors are presented.

Proteomic analysis reveals differential responsive mechanisms in Solanum nigrum exposed to low and high dose of cadmium.

Cadmium (Cd) contamination is becoming a widespread environmental problem. However, the differential responsive mechanisms of Cd hyperaccumulator Solanum nigrum to low or high dose of Cd are not well documented. In this study, phenotypic and physiological analysis firstly suggested that the seedlings of S. nigrum showed slight leaf chlorosis symptoms under 25 µM Cd and severe inhibition on growth and photosynthesis under 100 µM Cd. Further proteomic analysis identified 105 differentially expressed proteins (DEPs) in the Cd-treated leaves. Under low dose of Cd stress, 47 DEPs are mainly involved in primary metabolic processes, while under high dose of Cd stress, 92 DEPs are mainly involved in photosynthesis, energy metabolism, production of phytochelatin and reactive oxygen species (ROS). Protein-protein interaction (PPI) network analysis of DEPs support above differential responses in the leaves of S. nigrum to low and high dose of Cd treatments. This work provides the differential responsive mechanisms in S. nigrum to low and high dose of Cd, and the theoretical foundation for the application of hyperaccumulating plants in the phytoremediation of Cd-contaminated soils.