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1.
Cell ; 187(6): 1387-1401.e13, 2024 Mar 14.
Article in English | MEDLINE | ID: mdl-38412859

ABSTRACT

The Crumbs homolog 1 (CRB1) gene is associated with retinal degeneration, most commonly Leber congenital amaurosis (LCA) and retinitis pigmentosa (RP). Here, we demonstrate that murine retinas bearing the Rd8 mutation of Crb1 are characterized by the presence of intralesional bacteria. While normal CRB1 expression was enriched in the apical junctional complexes of retinal pigment epithelium and colonic enterocytes, Crb1 mutations dampened its expression at both sites. Consequent impairment of the outer blood retinal barrier and colonic intestinal epithelial barrier in Rd8 mice led to the translocation of intestinal bacteria from the lower gastrointestinal (GI) tract to the retina, resulting in secondary retinal degeneration. Either the depletion of bacteria systemically or the reintroduction of normal Crb1 expression colonically rescued Rd8-mutation-associated retinal degeneration without reversing the retinal barrier breach. Our data elucidate the pathogenesis of Crb1-mutation-associated retinal degenerations and suggest that antimicrobial agents have the potential to treat this devastating blinding disease.


Subject(s)
Nerve Tissue Proteins , Retinal Degeneration , Animals , Mice , Bacterial Translocation , Eye Proteins/genetics , Leber Congenital Amaurosis/genetics , Mutation , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Retina/metabolism , Retinal Degeneration/genetics , Retinitis Pigmentosa/genetics , Retinitis Pigmentosa/metabolism , Retinitis Pigmentosa/pathology
2.
Hum Mol Genet ; 33(15): 1356-1366, 2024 Jul 22.
Article in English | MEDLINE | ID: mdl-38723288

ABSTRACT

microRNAs (miRNAs) are short non-coding RNAs that have been increasingly recognized for their significant roles in the progression of cancer. Distinct miRNAs exhibit diverse functions attributed to variations in their sequences. As a result of possessing highly homologous seed sequences, these miRNAs target overlapping or similar gene sets, thus performing analogous roles. However, different from this sight, our study discovered that miR-135a-5p and miR-135b-5p, despite differing by only one nucleotide, exhibit distinct functional roles. Using non-small cell lung cancer (NSCLC) as a paradigm, our findings unveiled the downregulation of miR-135a-5p and upregulation of miR-135b-5p within NSCLC through TCGA database. Consequently, we further investigated their functional differences in A549 cells. Overexpression of miR-135b-5p enhanced the proliferation and migration capabilities of A549 cells, whereas miR-135a-5p transfection exhibited the opposite effect. We demonstrated that the activation of specific enhancers serves as a crucial mechanism underlying the disparate functions exerted by miR-135a-5p and miR-135b-5p in the context of NSCLC, consequently instigating a shift from inhibition to activation in NSCLC progression. Finally, we validated through animal experiments that miR-135b-5p promoted tumor progression, while miR-135a-5p exerted inhibitory effects on NSCLC development. This study offers a novel perspective for researchers to elucidate functional disparities exhibited by highly homologous miRNAs (miR-135a-5p and miR-135b-5p) in the context of NSCLC, along with the transition from inhibitory to progressive states in NSCLC. This study provides a solid foundation for future investigations into the functional roles of highly homologous miRNAs in pathological situation.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Cell Proliferation , Gene Expression Regulation, Neoplastic , Lung Neoplasms , MicroRNAs , MicroRNAs/genetics , Humans , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/pathology , Lung Neoplasms/genetics , Lung Neoplasms/pathology , Animals , Cell Proliferation/genetics , Mice , Disease Progression , Cell Movement/genetics , A549 Cells , Enhancer Elements, Genetic , Cell Line, Tumor
3.
Hum Mol Genet ; 31(12): 2035-2048, 2022 06 22.
Article in English | MEDLINE | ID: mdl-35022748

ABSTRACT

K63-linked polyubiquitination of the neurodegenerative disease-associated misfolded protein copper-zinc superoxide dismutase 1 (SOD1) is associated with the formation of inclusion bodies. Highly expressed E3 ligase Smad ubiquitylation regulatory factor 1 (Smurf1) promotes cellular homeostasis through the enhanced capability of aggregate degradation. However, it is not well explored the role of Smurf1 in the dynamics of SOD1 aggresomes. In this study, we report that Smurf1 promotes the recruitment of SOD1 to form aggresomes. Mechanistically, Smurf1 interacts with mutant SOD1 to promote aggresome formation by modification of its K63-linked polyubiquitination. Moreover, overexpressed Smurf1 enhances mutant SOD1 aggresome formation and autophagic degradation to prevent cell death. Thus, our data suggest that Smurf1 plays an important role in attenuating protein misfolding-induced cell toxicity by both driving the sequestration of misfolded SOD1 into aggresomes and autophagic degradation.


Subject(s)
Neurodegenerative Diseases , Superoxide Dismutase-1 , Ubiquitin-Protein Ligases , Humans , Inclusion Bodies/metabolism , Neurodegenerative Diseases/metabolism , Protein Folding , Superoxide Dismutase-1/genetics , Superoxide Dismutase-1/metabolism , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism , Ubiquitination
4.
J Transl Med ; 22(1): 474, 2024 May 19.
Article in English | MEDLINE | ID: mdl-38764020

ABSTRACT

BACKGROUND: The initiation of fibroblast growth factor 1 (FGF1) expression coincident with the decrease of FGF2 expression is a well-documented event in prostate cancer (PCa) progression. Lactate dehydrogenase A (LDHA) and LDHB are essential metabolic products that promote tumor growth. However, the relationship between FGF1/FGF2 and LDHA/B-mediated glycolysis in PCa progression is not reported. Thus, we aimed to explore whether FGF1/2 could regulate LDHA and LDHB to promote glycolysis and explored the involved signaling pathway in PCa progression. METHODS: In vitro studies used RT‒qPCR, Western blot, CCK-8 assays, and flow cytometry to analyze gene and protein expression, cell viability, apoptosis, and cell cycle in PCa cell lines. Glycolysis was assessed by measuring glucose consumption, lactate production, and extracellular acidification rate (ECAR). For in vivo studies, a xenograft mouse model of PCa was established and treated with an FGF pathway inhibitor, and tumor growth was monitored. RESULTS: FGF1, FGF2, and LDHA were expressed at high levels in PCa cells, while LDHB expression was low. FGF1/2 positively modulated LDHA and negatively modulated LDHB in PCa cells. The depletion of FGF1, FGF2, or LDHA reduced cell proliferation, induced cell cycle arrest, and inhibited glycolysis. LDHB overexpression showed similar inhibitory effect on PCa cells. Mechanistically, we found that FGF1/2 positively regulated STAT1 and STAT1 transcriptionally activated LDHA expression while suppressed LDHB expression. Furthermore, the treatment of an FGF pathway inhibitor suppressed PCa tumor growth in mice. CONCLUSION: The FGF pathway facilitates glycolysis by activating LDHA and suppressing LDHB in a STAT1-dependent manner in PCa.


Subject(s)
Fibroblast Growth Factors , Glycolysis , L-Lactate Dehydrogenase , Prostatic Neoplasms , STAT1 Transcription Factor , Signal Transduction , Male , Prostatic Neoplasms/pathology , Prostatic Neoplasms/metabolism , Prostatic Neoplasms/genetics , Humans , Animals , L-Lactate Dehydrogenase/metabolism , Cell Line, Tumor , STAT1 Transcription Factor/metabolism , Fibroblast Growth Factors/metabolism , Mice, Nude , Cell Proliferation , Mice , Gene Expression Regulation, Neoplastic , Fibroblast Growth Factor 2/metabolism , Apoptosis , Lactate Dehydrogenase 5/metabolism , Isoenzymes
5.
Anal Bioanal Chem ; 2024 Jul 13.
Article in English | MEDLINE | ID: mdl-38997460

ABSTRACT

Therapeutic drug monitoring is essential for ensuring the efficacy and safety of medications. This study introduces a streamlined approach that combines pipette-tip solid-phase extraction (PT-SPE) with matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS), facilitating rapid and high-throughput monitoring of drug concentrations. As a demonstration, this method was applied to the extraction and quantification of antidepressants in serum. Utilizing Zip-Tip C18, the method enabled the extraction of antidepressants from complex biological matrices in less than 2 min, with the subsequent MALDI-MS analysis yielding results in just 1 min. Optimal extraction recoveries were achieved using a sampling solution at pH 9.0 and a 10 µL ethanol desorption solution containing 0.1% phosphoric acid. For MALDI analysis, 2,5-dihydroxybenzoic acid was identified as the most effective matrix for producing the highest signal intensity. The quantification strategy exhibited robust linearities (R2 ≥ 0.997) and satisfactory limits of quantification, ranging from 0.05 to 0.5 µg/mL for a suite of antidepressants. The application for monitoring dynamic concentration changes of antidepressants in rat serum emphasized the method's efficacy. This strategy offers the advantages of high throughput, minimal sample usage, environmental sustainability, and simplicity, providing ideas and a reference basis for the subsequent development of methods for therapeutic drug monitoring.

6.
Phys Chem Chem Phys ; 26(29): 19822-19830, 2024 Jul 24.
Article in English | MEDLINE | ID: mdl-38988227

ABSTRACT

Single-atom catalysts (SACs) play a vital role in the hydrogen evolution reaction (HER) owing to the highly desirable atom efficiency and the minimal amount of precious metals. Herein, we use TiO2 nanosheets to anchor stable atomically dispersed iridium (Ir) to be used as a catalyst (Ir@TiO2) for the HER. The atomic dispersion of Ir on the TiO2 substrate is confirmed by aberration-corrected scanning transmission electron microscopy and it is anchored by numerous surface functional groups on abundantly exposed basal planes in TiO2. In acidic media, the Ir@TiO2 catalyst (1.35 wt% Ir) shows a low overpotential (41 mV at 10 mA cm-2), a small Tafel slope of 42 mV dec-1, and a decent durability for 1000 cycles of the HER with the polarization curve having only a 1 mV shift, which are comparable with those of a commercial Pt/C catalyst with 20 wt% Pt. This work paves a way to design Ir atomically anchored catalysts with low cost and high activity.

7.
Ecotoxicol Environ Saf ; 275: 116225, 2024 Apr 15.
Article in English | MEDLINE | ID: mdl-38520810

ABSTRACT

The honeycomb magnetic carbons (xFe@HCNs) were prepared by sacrificial template method novelty using polyacrylamide resin (PAAS) as template and ammonium pyrrolidine dithioate/Fe3+ complex (APDC-Fe) as carbon skeleton and metal source. Tetracycline (TC) and copper (Cu2+) as target pollutants were used to investigate the adsorption properties of xFe@HCNs in single or binary TC and Cu2+ systems. The adsorption capacity sequence for TC among the adsorbents was (mmol·g-1): 2Fe@HCNs (0.088) > 8Fe@HCNs (0.061) > HCNs (0.054) > RC (0.036), and for Cu2+ was (mmol·g-1): 2Fe@HCNs (1.120) > 8Fe@HCNs (1.026) > RC (0.792) > HCNs (0.681). 2Fe@HCNs demonstrated notable affinity for adsorbing both TC and Cu2+. Additionally, the influence of hydrochemical factors (i.e., cation species, anion species, and pH) on the adsorption properties of 2Fe@HCNs. Combined with advanced oxidation technology, the regeneration methods of magnetic adsorbent were explored using oxidizing agents (e.g., H2O2 and peroxymonosulfate) as eluents which could increase the adsorption sites of magnetic carbon adsorbents during the regenerating process, which was the novelty of the study. Furthermore, the regeneration mechanisms of H2O2 as eluent were investigated. This study discussed the application and regeneration methods of magnetic adsorbents in water treatment, offering new insights into environmental remediation using magnetic materials.


Subject(s)
Heterocyclic Compounds , Water Pollutants, Chemical , Carbon/chemistry , Copper/chemistry , Adsorption , Hydrogen Peroxide , Water Pollutants, Chemical/chemistry , Tetracycline/chemistry , Anti-Bacterial Agents , Magnetic Phenomena , Kinetics
8.
Genomics ; 115(4): 110660, 2023 Jul.
Article in English | MEDLINE | ID: mdl-37257521

ABSTRACT

Canine mammary tumor (CMT) are the second most common tumor in dogs. Exosomes can act as biomarkers for the early diagnosis of tumors, and also be involved in the pathogenesis and metastasis mechanism of tumors. The expression profile of exosomal RNA revealed that there were a total of 5547 differentially expressed mRNAs, and 196 differentially expressed lncRNAs. GO and KEGG enrichment analysis found that the differentially expressed mRNAs and lncRNA target genes were associated with metabolic processes, DNA replication, cell proliferation, cell junction, and cell adhesion. In conclusion, this study revealed lncRNA and mRNA expression profiles in exosomes derived from plasma of CMT and further annotated their potential functions. The data obtained in this study will also provide valuable resources for understanding lncRNA information in plasma exosomes of dogs with CMT, and contribute to the study of early diagnostic markers and pathogenesis of CMT.


Subject(s)
Exosomes , RNA, Long Noncoding , Dogs , Animals , Gene Expression Profiling , Exosomes/genetics , Exosomes/metabolism , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Gene Regulatory Networks , RNA-Seq
9.
Int J Mol Sci ; 25(14)2024 Jul 10.
Article in English | MEDLINE | ID: mdl-39062806

ABSTRACT

Histone arginine residue methylation is crucial for individual development and gene regulation. However, the dynamics of histone arginine methylation in response to cellular stress remains largely unexplored. In addition, the interplay and regulatory mechanisms between this and other histone modifications are important scientific questions that require further investigation. This study aimed to investigate the changes in histone arginine methylation in response to DNA damage. We report a global decrease in histone H3R26 symmetric dimethylation (H3R26me2s) and hypoacetylation at the H3K27 site in response to DNA damage. Notably, H3R26me2s exhibits a distribution pattern similar to that of H3K27ac across the genome, both of which are antagonistic to H3K27me3. Additionally, histone deacetylase 1 (HDAC1) may be recruited to the H3R26me2s demethylation region to mediate H3K27 deacetylation. These findings suggest crosstalk between H3R26me2s and H3K27ac in regulating gene expression.


Subject(s)
Arginine , Histone Deacetylase 1 , Histones , Histones/metabolism , Arginine/metabolism , Methylation , Humans , Histone Deacetylase 1/metabolism , Histone Deacetylase 1/genetics , DNA Damage , Acetylation , Protein Processing, Post-Translational , Stress, Physiological/genetics
10.
Water Sci Technol ; 89(5): 1179-1194, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38483492

ABSTRACT

The study investigated the denitrification effect of the iron autotrophic denitrification process for removing nitrite under anaerobic conditions, utilizing sponge iron as the electron donor. When the C/N ratio equaled 1, defined as the ratio of chemical oxygen demand to total nitrogen (TN), and the influent nitrite nitrogen (NO2--N) was at 80 mg/L, the average steady-state TN effluent concentration of this system was 41.94 mg/L during the 79-day experiment. The TN value exhibited a significant decrease compared to both the sponge iron system (68.69 mg/L) and the carbon source system (56.50 mg/L). Sponge iron is beneficial for providing an electron donor and ensuring an anaerobic system, fostering an environment that promotes microorganism growth while effectively inhibiting the conversion of nitrite to nitrate. In addition, carbon sources play a vital role in ensuring microorganism growth and reproduction, thereby aiding in TN removal. The optimal parameters based on the effectiveness of TN removal in the iron autotrophic denitrification system were determined to be s-Fe0 dosage of 30 g/L and C/N = 1.5. These results suggest that the iron autotrophic denitrification process, driven by sponge iron, can effectively remove nitrite under anaerobic conditions.


Subject(s)
Denitrification , Nitrites , Anaerobiosis , Bioreactors , Carbon , Iron , Nitrogen
11.
Langmuir ; 39(35): 12510-12519, 2023 Sep 05.
Article in English | MEDLINE | ID: mdl-37667672

ABSTRACT

As is known, metal-organic frameworks (MOFs) are a versatile class of materials in energy storage applications including supercapacitors. However, the individual kind of metal nodes connected by organic ligands to form a topological structure still limits the potential storage capacity of MOFs. Herein, a bimetal-based Ni-Mn MOF composite is configured with a one-pot hydrothermal method to derive a composite with a synergic effect to maximize the properties. Moreover, reduced graphene oxide (rGO) sheets are added as a conductive network to anchor the MOF-derived composite of Ni-Mn@C/rGO, which is expected to increase the conductivity of the materials system. The resulting composite exhibited a high specific capacitance of 1674 F g-1 at a current density of 0.3 A g-1, suggesting excellent energy storage performance. The composite was then integrated as the cathode in an asymmetrical supercapacitor with a 3D rGO aerogel anode, resulting in energy densities of 24.1 and 17.5 W h kg-1 at power densities of 88.9 and 444.4 W kg-1, respectively. Additionally, the device demonstrated remarkable long-term stability, with 90% capacitance retention after 10 000 charge-discharge cycles at 10 A g-1.

12.
J Org Chem ; 88(6): 3981-3986, 2023 Mar 17.
Article in English | MEDLINE | ID: mdl-36862134

ABSTRACT

The total synthesis of levesquamide, a natural product with an unprecedented pentasubstituted pyridine-isothiazolinone skeleton, has been accomplished from kojic acid for the first time. The key features of the synthesis include a Suzuki coupling reaction between bromopyranone and oxazolyl borate fragments, a copper-mediated introduction of a thioether, a mild hydrolysis of a pyridine 2-N-methoxyamide, and a Pummerer-type cyclization of a tert-butyl sulfoxide to form the key pyridine-isothiazolinone unit of the natural product.

13.
Neurol Sci ; 44(6): 1891-1903, 2023 Jun.
Article in English | MEDLINE | ID: mdl-36781567

ABSTRACT

BACKGROUND: Cognitive impairment was a common sequela among stroke survivors, and exercise intervention was a promising non-pharmacological treatment modality for it. PURPOSE: To explore the effects of exercise intervention programs on cognitive and motor function in patients with cognitive impairment after stroke. STUDY DESIGN: Systematic review and meta-analysis. METHODS: Seven online databases (PubMed, Embase, Cochrane Library, Web of Science, Scopus, PsycInfo, and SPORTDiscus) were searched from their inception to 10 February 2022. Randomised controlled trials (RCTs) comparing the effects of exercise with non-exercise rehabilitation, using the Montreal Cognitive Assessment, Addenbrooke's Cognitive Examination, Mini-Mental State Examination, Trial Making Test, Upper and Lower Extremity Fugl-Meyer Assessment, Berg Balance Scale, and Barthel Index, were selected. Calculations for each assessment were performed for the overall effect and the therapy of interest, taking into account the effect of stroke severity or stimulus parameters. RESULTS: Twelve RCTs involving 975 participants and investigating nine different types of exercise interventions were included. The results were not affected by participant characteristics or reactive balance outcomes. Our results emphasise the importance of lightweight and operable aerobic exercises. Exercise itself had a high potential to improve cognitive impairment and motor function after stroke. CONCLUSIONS: Exercise had significant positive effects on alleviating cognitive and motor impairments after stroke.


Subject(s)
Stroke Rehabilitation , Stroke , Humans , Stroke Rehabilitation/methods , Stroke/complications , Stroke/therapy , Cognition/physiology , Exercise Therapy , Survivors
14.
Nucleic Acids Res ; 49(15): 8556-8572, 2021 09 07.
Article in English | MEDLINE | ID: mdl-34329471

ABSTRACT

Dysfunction of Tumour Suppressor Genes (TSGs) is a common feature in carcinogenesis. Epigenetic abnormalities including DNA hypermethylation or aberrant histone modifications in promoter regions have been described for interpreting TSG inactivation. However, in many instances, how TSGs are silenced in tumours are largely unknown. Given that miRNA with low expression in tumours is another recognized signature, we hypothesize that low expression of miRNA may reduce the activity of TSG related enhancers and further lead to inactivation of TSG during cancer development. Here, we reported that low expression of miRNA in cancer as a recognized signature leads to loss of function of TSGs in breast cancer. In 157 paired breast cancer and adjacent normal samples, tumour suppressor gene GPER1 and miR-339 are both downregulated in Luminal A/B and Triple Negative Breast Cancer subtypes. Mechanistic investigations revealed that miR-339 upregulates GPER1 expression in breast cancer cells by switching on the GPER1 enhancer, which can be blocked by enhancer deletion through the CRISPR/Cas9 system. Collectively, our findings reveal novel mechanistic insights into TSG dysfunction in cancer development, and provide evidence that reactivation of TSG by enhancer switching may be a promising alternative strategy for clinical breast cancer treatment.


Subject(s)
Breast Neoplasms/genetics , DNA Methylation/genetics , MicroRNAs/genetics , Receptors, Estrogen/genetics , Receptors, G-Protein-Coupled/genetics , Tumor Suppressor Proteins/genetics , Breast Neoplasms/pathology , Carcinogenesis/genetics , Enhancer Elements, Genetic/genetics , Epigenomics , Female , Gene Expression Regulation, Neoplastic/genetics , Gene Silencing , Humans , Promoter Regions, Genetic/genetics , RNA, Neoplasm/genetics , Regulatory Sequences, Nucleic Acid/genetics
15.
Ecotoxicol Environ Saf ; 253: 114668, 2023 Mar 15.
Article in English | MEDLINE | ID: mdl-36812870

ABSTRACT

Antibiotics are omnipresent and pseudo-persistent in the environment. Yet, their potential ecological risks under repeated exposure, which is more environmentally relevant, are understudied. Therefore, this study used ofloxacin (OFL) as the probe chemical to investigate the toxic effects of different exposure scenarios-single dose of high concentration (4.0 µg/L) and multiple additions of low concentrations-towards the cyanobacterium Microcystis aeruginosa. Flow cytometry was employed to measure a collection of biomarkers, including endpoints related with biomass, single cell properties and physiological status. Results showed that the single dose of the highest OFL level inhibited cellular growth, chl-a content and cell size of M. aeruginosa. In contrast, OFL induced stronger chl-a autofluorescence and higher doses tended to have more remarkable effects. Repeated low OFL doses can more significantly increase the metabolic activity of M. aeruginosa than a single high dose. Viability and cytoplasmic membrane were not affected by OFL exposure. Oxidative stress was observed for the different exposure scenarios, with fluctuating responses. This study demonstrated the different physiological responses of M. aeruginosa under different OFL exposure scenarios, providing novel insights into the toxicity of antibiotics under repeated exposure.


Subject(s)
Microcystis , Ofloxacin , Ofloxacin/toxicity , Ofloxacin/metabolism , Anti-Bacterial Agents/pharmacology , Oxidative Stress
16.
Ecotoxicol Environ Saf ; 263: 115214, 2023 Sep 15.
Article in English | MEDLINE | ID: mdl-37413944

ABSTRACT

Brassica campestris L., a cadmium (Cd) hyperaccumulating herbaceous plant, is considered as a promising candidate for the bioremediation of Cd pollution. However, the molecular mechanisms regulating these processes remain unclear. The present work, using proteome studies combined with a transcriptome analysis, was carried out to reveal the response mechanisms of the hairy roots of Brassica campestris L. under Cd stress. Significant tissue necrosis and cellular damage occurred, and Cd accumulation was observed in the cell walls and vacuoles of the hairy roots. Through quantitative proteomic profiling, a total of 1424 differentially expressed proteins (DEPs) were identified, and are known to be enriched in processes including phenylalanine metabolism, plant hormone signal transduction, cysteine and methionine metabolism, protein export, isoquinoline alkaloid biosynthesis and flavone biosynthesis. Further studies combined with a transcriptome analysis found that 118 differentially expressed genes (DEGs) and their corresponding proteins were simultaneously up- or downregulated. Further Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analysis of the 118 shared DEGs and DEPs indicated their involvement in calcium, ROS and hormone signaling-mediated response, including regulation of carbohydrate and energy metabolism, biosynthesis of GSH, PCs and phenylpropanoid compounds that play vital roles in the Cd tolerance of Brassica campestris L. Our findings contribute to a better understanding of the regulatory networks of Brassica campestris L. under Cd stress, as well as provide valuable information on candidate genes (e.g., BrPAL, BrTAT, Br4CL, BrCDPK, BrRBOH, BrCALM, BrABCG1/2, BrVIP, BrGCLC, BrilvE, BrGST12/13/25). These results are of particular importance to the subsequent development of promising transgenic plants that will hyperaccumulate heavy metals and efficient phytoremediation processes.


Subject(s)
Brassica , Cadmium , Cadmium/toxicity , Cadmium/metabolism , Brassica/metabolism , Proteome/metabolism , Proteomics , Stress, Physiological/genetics , Gene Expression Profiling/methods , Metabolic Networks and Pathways/genetics , Transcriptome , Plant Roots/genetics , Plant Roots/metabolism
17.
Int J Phytoremediation ; 25(11): 1455-1462, 2023.
Article in English | MEDLINE | ID: mdl-36597829

ABSTRACT

Cadmium (Cd) is the main heavy metal pollutant in soil. The combination of genetic engineering technology and Rizobium rhizogenes mediated technology can effectively improve the enrichment efficiency of heavy metals in super accumulators and reduce soil heavy metal pollution. In this study, the transgenic hairy root system containing the IRT1 gene of Cd hyperaccumulator-Brassica campestris L. was successfully constructed by the R. rhizogenes mediated method (IRT1 gene come from Arabidopsis thaliana). The hairy roots of each subculture can grow stably within 6 weeks, and IRT1 gene will not be lost within 50 subcultures., which is detected using PCR method. The results of Cd enrichment experiments showed that after treatment with 100 µmol/L Cd for 14 days, the growth state of transgenic IRT1 hairy roots only showed slight browning. Also, the accumulation value of Cd reached 331.61 µg/g and the enrichment efficiency of transgenic IRT1 hairy roots was 13.8% higher than that of wild-type hairy roots. Western blotting results showed that the expression of IRT1 protein in transgenic hairy roots was significantly higher than that of wild-type hairy roots under Cd stress. The above results indicated that the overexpression of IRT1 gene can help B. campestris L. hairy roots to effectively cope with Cd stress and improve its ability to enrich Cd.


In this study, the transgenic hairy root system containing the IRT1 gene of Cd hyperaccumulator-Brassica campestris L. was successfully constructed by the Rizobium rhizogenes mediated method. At the same time, the growth state and cadmium enrichment efficiency of transgenic hairy roots under different concentrations of Cd stress were studied. Overexpression of IRT1 gene can effectively improve the tolerance of hairy root to Cd. The enrichment efficiency of transgenic IRT1 hairy roots was 13.8% higher than that of wild-type hairy roots. The transgenic IRT1 hairy root system established in this study can be used as a reliable experimental model for the study of Cd adsorption mechanism, and can be further regenerated to obtain transgenic IRT1 B. campestris L. plants for the study of heavy metal Cd pollution remediation.


Subject(s)
Arabidopsis Proteins , Arabidopsis , Brassica , Cation Transport Proteins , Brassica/genetics , Brassica/metabolism , Cadmium/metabolism , Biodegradation, Environmental , Arabidopsis/genetics , Arabidopsis/metabolism , Soil , Plant Roots , Cation Transport Proteins/metabolism , Arabidopsis Proteins/metabolism
18.
Int Orthop ; 46(9): 2019-2028, 2022 09.
Article in English | MEDLINE | ID: mdl-35616654

ABSTRACT

PURPOSE: We developed an augmentation technique for PCL reconstruction with independent internal brace reinforcement and evaluated the functional outcome after PCL reconstruction employing autologous hamstrings augmented with an internal brace system for patients with isolated or combined grade 3 posterior instability who were treated with this technique. METHODS: From January 2016 to January 2018, patients with isolated or combined grade 3 PCL tears who underwent single-bundle PCL reconstruction using autologous hamstrings augmented with independent internal braces were studied. The function of the operated knee was evaluated according to the International Knee Documentation Committee (IKDC) score, Lysholm score, and Tegner activity score. The patients were asked the level of returned to their previous sport. Posterior knee laxity was examined with a KT-1000 arthrometer, and data on range of motion (ROM), re-operation, and other complications were collected. RESULTS: A total of 33 consecutive patients who received single-bundle PCL reconstruction using autologous hamstrings augmented with independent internal braces with a minimum two years follow-up were included in this study. Two patients had undergone this procedure during the study period and were not included in this study (one had combined bone fractures, and one patient had previous meniscus surgery). Thirty-one patients were available for final analysis. The mean follow-up was 45.35 ± 10.88 months (range 29-66 months). The average IKDC subjective knee evaluation scores from 51.65 ± 12.35 to 84.52 ± 6.42, the Lysholm score from 53.90 ± 11.86 to 85.68 ± 4.99, and the Tegner score from 2.81 ± 0.79 to 6.71 ± 1.83 (P < 0.05 for all). The mean total posterior side-to-side difference in knee laxity, assessed using a KT-1000 arthrometer, decreased from 12.13 ± 2.66 mm pre-operatively to 1.87 ± 0.56 mm post-operatively at 70° (P < 0.05). Most patients (29/31) had normal or near normal knee ROM post-operatively; two patients revealed a 6-15° loss of knee flexion compared with the contralateral knee. Twenty-nine patients (93.55%) returned to a normal daily exercise level. Twenty-three patients (74.19%) returned to competitive sports with high-level sports (Tegner score of 6 or above; eleven patients (35.48%) reported to be on the same level as well as the Tegner level); six patients (19.35%) returned to recreational sports (Tegner score of 4 or 5). Two patients had Tegner scores of 2 and 3, indicating poor function level. No patient needed PCL revision surgery during the follow-up period. CONCLUSION: Single-bundle PCL reconstruction with internal brace augmentation for PCL injury exhibited satisfactory posterior stability and clinical outcomes in patients with isolated or combined grade 3 PCL injuries at a minimum two year follow-up.


Subject(s)
Joint Instability , Knee Injuries , Posterior Cruciate Ligament Reconstruction , Posterior Cruciate Ligament , Follow-Up Studies , Humans , Joint Instability/etiology , Joint Instability/surgery , Knee Injuries/surgery , Knee Joint/surgery , Posterior Cruciate Ligament/injuries , Posterior Cruciate Ligament/surgery , Treatment Outcome
19.
J Environ Manage ; 311: 114889, 2022 Mar 11.
Article in English | MEDLINE | ID: mdl-35287073

ABSTRACT

Cyanobacterial blooms challenge the safe water supply in estuary reservoirs. Yet, data are limited for the variation of phytoplankton dynamics during an algal bloom event at refined scales, which is essential for interpreting the formation and cessation of blooms. The present study investigated the biweekly abundances and dynamics of pico- and nano-phytoplankton in a tropical estuary lake following a prolonged bloom event. Flow cytometry analysis resolved eight phenotypically distinct groups of phytoplankton assigned to nano-eukaryotes (nano-EU), pico/nano-eukaryotes (PicoNano-EU), cryptophyte-like cells (CRPTO), Microcystis-like cells (MIC), pico-eukaryotes (Pico-EU) and three groups of Synechococcus-like cells. Total phytoplankton abundance ranged widely from 2.4 × 104 to 2.8 × 106 cells cm-3. The phytoplankton community was dominated by Synechococcus-like cells with high phycocyanin content (SYN-PC). Temporal dynamics of the phytoplankton community was phytoplankton- and site-specific. Peak values were observed for SYN-PC, SYN-PE2 (Synechococcus-like cells with low levels of phycoerythrin) and Pico-EU, while the temporal dynamics of other groups were less pronounced. Redundancy analysis (RDA) showed the importance of turbidity as an abiotic factor in the formation of the current SYN-PC induced blooms, and Spearman correlation analysis suggested a competitive relationship between SYN-PC and Pico-EU.

20.
J Environ Manage ; 322: 116086, 2022 Nov 15.
Article in English | MEDLINE | ID: mdl-36041306

ABSTRACT

The application of anaerobic ammonium oxidation (Anammox) technology in low-strength wastewater treatment still faces difficult in-situ start-ups and unstable operations. Sponge-iron sludge (R1) was used as a novel inoculum to provide a promising solution. Conventional activated sludge (R0) was used as the control. However, little is known about the feasibility and performance during the start-up and operation of Anammox combined with biological iron and iron bacteria in an iron sludge system. Anammox was successfully started both in R1 (87 days) and R0 (89 days) with a low-strength influent (with a nitrogen loading rate (NLR) of 43.64 ± 0.41 g N/(m3⋅d)). During long-term operation, the R0 nevertheless produced higher nitrates (9.7 ± 0.1 mg/L) than expected. In contrast, R1 presented no excess nitrate production (2.1 ± 0.06 mg/L). The total inorganic nitrogen (TIN) removal efficiency increased from 78.2 ± 7.1% in R0 to 86.1 ± 4.3% in R1. The iron sludge in R1 was divided equally into three parts and three different nitrogen-feeding methods were used over the 34 days of operation, as follows: first using a mixture of ammonium (27.15 ± 1.0 mg/L) and nitrite (32.7 ± 1.7 mg/L), then only ammonium (27.15 ± 1.0 mg/L) and lastly only nitrite (32.7 ± 1.7 mg/L) as the influent. R1 was a coupled system composed of Anammox, Feammox, and NOx--dependent Fe(II) oxidation (NDFO). The contribution of Feammox and NDFO to TIN removal was 27.1 ± 1.2% and 31.9 ± 0.7%. However, Anammox was the primary nitrogen transformation pathway. X-ray diffraction (XRD) analysis shows that iron hydroxide (Fe(OH)3) and iron oxide hydroxide (FeOOH) were generated in R1. The produced Fe(OH)3 and FeOOH were capable of participating in Feammox and formed a Fe(II)/Fe(III) cycle which further removed nitrogen. Therefore, a highly stable and impressive nitrogen removal performance was demonstrated in the iron sludge Anammox system under the cooperation of biological iron and iron bacteria. The study considered the enrichment of norank_c_OM190, Desulfuromonas, and Thiobacillus and their contribution to the Anammox, Feammox, and NDFO processes, respectively. This study provides a new perspective for the start-up and stable operation of low-strength wastewater Anammox engineering applications.


Subject(s)
Ammonium Compounds , Sewage , Bacteria/metabolism , Bioreactors/microbiology , Denitrification , Ferric Compounds , Ferrous Compounds , Iron , Nitrates , Nitrites , Nitrogen/metabolism , Oxidation-Reduction , Sewage/microbiology , Wastewater/microbiology
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