ENVIRONMENTAL SCIENCE & TECHNOLOGY
·
2022
·
IF 11.357
Jieyi Zeng,Yaying Li,Guoping Jin,Jian-Qiang Su,Huaiying Yao
Antibiotic resistance genes (ARGs) are global pollutants that pose a potential risk to human health. Benzalkonium chloride (C12) (BC) disinfectants are thought to exert selection pressure on antibiotic resistance. However, evidence of BC-induced changes in antibiotic resistance in the soil environment is lacking. Here, we established short-term soil microcosms to investigate ARG profile dynamics in agricultural soils amended with sulfamethazine (SMZ, 10 mg kg–1) and gradient concentrations of BC (0–100 mg kg–1), using high-throughput quantitative PCR and Illumina sequencing. With the increase in BC concentration, the number of ARGs detected in the soil increased, but the normalized ARG abundance decreased. The added SMZ had a limited impact on ARG profiles. Compared to broad-spectrum fungicidal BC, the specificity of SMZ significantly affected the microbial community. Network analysis found that low–medium BC exposure concentrations resulted in the formation of small but strong ARG co-occurrence clusters in the soil, while high BC exposure concentration led to a higher incidence of ARGs. Variation partitioning analysis suggested that BC stress was the major driver shaping the ARG profile. Overall, this study highlighted the emergence and spread of BC-induced ARGs, potentially leading to the antimicrobial resistance problem in agricultural soils.
研究领域:
环境科学
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微生物学
·
土壤学
·
基因组学
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产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2021
·
IF 6.558
Feixia Peng,Mei Liu,Xuelian Wang,Xueqin Ding
Fast extraction of high-purity nucleic acid from complex biological sample is the key to downstream nucleic acid analysis. In this work, two low-viscosity hydrophobic magnetic deep eutectic solvents (HMDESs) were synthesized for the selective extraction of DNA . The conformation of the HMDES was simulated by density functional theory (DFT) calculation. Characterization of HMDESs’ physical (magnetism, density, viscosity, and hydrophobicity) properties and thermal (melting point and decomposition temperature) properties were conducted. Based on the HMDESs, a vortex-assisted liquid-liquid micro-extraction (VALLME) DNA method was developed. Single stranded DNA that was extracted by HMDESs could be quickly collected by an external magnet. Factors that could impact the DNA extraction process, such as HMDESs volume, temperature, extraction time, and pH were systematically investigated via single-factor experimental analysis. Under the optimized condition, the proposed extraction method has been demonstrated with the extraction of DNA from a series of complex sample matrices, including metal mixture, protein mixture and E. coli cell lysate. The DNA extracted by using HMDES-based VALLME method was well suitable for PCR amplifications. After extraction, the retained DNAs could be readily recovered by simply using Britton-Robison (BR) buffer. In addition, the interaction and corresponding binding sites between HMDESs and DNA were investigated by FT-IR and DFT calculation. This work provides a new green magnetic solvent and a rapid and environmental-friendly extraction method for the enrichment of DNA and other biological macromolecules .
研究领域:
分析化学
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分子生物学
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生物技术
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纳米技术
✓ 已验证
DOI
PubMed
高影响因子
American Journal of Translational Research
·
2021
·Vol. 13
·期 4
·2111-2126
·
IF 4.06
Chao Shi,Chongwen Shen,Ge Liu,Shaofeng Yang,Fenglin Ye,Jinjin Meng,Youmin Pan
Purpose To clarify the regulatory effect of Nuclear-enriched abundant transcript 1 (NEAT1) on abdominal aortic aneurysm (AAA) model rats and isolated endothelial progenitor cells (EPCs). Methods The AAA rat model was established by CaCl 2 stimulation, and overexpressed NEAT1 was injected into rats through tail vein. Abdominal aorta lesions and numbers of EPCs in tissues and peripheral blood were examined by hematoxylin-eosin, immunofluorescence and flow cytometry. The extracted EPCs were identified by microscopy, DiI-ac-LDL staining and flow cytometry. Effect of overexpressed/silencing NEAT1 on the viability, migration, tube formation and VEGF content of EPCs was investigated by MTT-, wound-healing, tube formation assays and ELISA, respectively. The expressions of NEAT1, miR-204-5p, Angiopoietin-1 (Ang-1)/ERK pathway were determined by qRT-PCR and Western blot as needed. The targeting relationships between NEAT1 and miR-204-5p, and miR-204-5p and Ang-1 were predicted on starBase, TargetScan and confirmed by dual-luciferase experiments. The mutual regulation effect was studied through rescue experiments. Results Overexpressed NEAT1 not only reduced inflammatory infiltration and increased the number of EPCs in abdominal aorta and peripheral blood, but also promoted the viability, migration, tube formation of EPCs, increased VEGF content and upregulated the expression of the Ang-1/ERK pathway in EPCs. However, silencing NEAT1 produced opposite results. NEAT1 targeting miR-204-5p inhibited the functional effects of miR-204-5p on of EPCs. Overexpressed/silencing Ang-1 partially reversed the effects of NEAT1 or miR-204-5p on the characteristics of EPCs. Conclusion NEAT1 competitively binds with miR-204-5p and up-regulates Ang-1 expression in EPCs to effectively improve the proliferation, migration and angiogenesis of EPCs.
研究领域:
分子生物学
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心血管疾病
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细胞生物学
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基因调控
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产品关联
DOI
PubMed
高影响因子
RUSSIAN JOURNAL OF PLANT PHYSIOLOGY
·
2018
·Vol. 65
·期 3
·372-383
·
IF 0.816
Zhang Y.,Yong Y. B.,Wang Q.,Lu Y. M.
Lily is regarded as one kind of the most economically important cut flowers in China. In this paper, lily cultivar ‘Aladdin’ ( Lilium longiflorum × Lilium asiatic ) was chosen as experimental material for its distinct high propagation coefficient in nature and three main conclusions were achieved. Firstly, our observation results revealed that the noticeable propagation coefficient of ‘Aladdin’ depended on strong capacity to form many underground stem axillary bulbils positioned on underground node of plant after 70 days of cultivation and having 1 cm flower buds. Secondly, in terms of physiological changes, zeatin riboside (ZR) relative concentration, soluble sugar and starch content peak when and where such bulbils are forming. Finally, two KNOX family Lilium genes were sequenced completely and one BELL family gene was sequenced partly; besides, there might be an interaction between KNOX-2 and BELL protein to accomplish their functions in bulbils organogenesis according to qRT-PCR results. As far as we know, it is the first time to study the formation mechanism of lily underground stem axillary bulbils. We illustrated physiological and molecular profiles, as well as offered a new prospect to lily molecular breeding in the future.
研究领域:
植物学
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分子生物学
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植物生理学
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遗传学
✓ 已验证
DOI
高影响因子
Parasites & Vectors
·
2021
·Vol. 14
·期 1
·1-10
·
IF 3.876
Gao Hai-Jun,Sun Xu-Dong,Luo Yan-Ping,Pang Hua-Sheng,Ma Xing-Ming,Zhang Ting,Jing Tao,Hu Wei,Shen Yu-Juan,Cao Jian-Ping
Background Echinococcosis, which is caused by the larvae of cestodes of the genus Echinococcus , is a parasitic zoonosis that poses a serious threat to the health of humans and animals globally. Albendazole is the drug of choice for the treatment of echinococcosis, but it is difficult to meet clinical goals with this chemotherapy due to its low cure rate and associated side effects after its long-term use. Hence, novel anti-parasitic targets and effective treatment alternatives are urgently needed. A previous study showed that verapamil (Vepm) can suppress the growth of Echinococcus granulosus larvae; however, the mechanism of this effect remains unclear. The aim of the present study was to gain insight into the anti-echinococcal effect of Vepm on Echinococcus with a particular focus on the regulatory effect of Vepm on calcium/calmodulin-dependent protein kinase II (Ca 2+ /CaM-CaMKII) in infected mice. Methods The anti-echinococcal effects of Vepm on Echinococcus granulosus protoscoleces (PSC) in vitro and Echinococcus multilocularis metacestodes in infected mice were assessed. The morphological alterations in Echinococcus spp. induced by Vepm were observed by scanning electron microscopy (SEM), and the changes in calcium content in both the parasite and mouse serum and liver were measured by SEM-energy dispersive spectrometry, inductively coupled plasma mass spectrometry and alizarin red staining. Additionally, the changes in the protein and mRNA levels of CaM and CaMKII in infected mice, and in the mRNA levels of CaMKII in E. granulosus PSC, were evaluated after treatment with Vepm by immunohistochemistry and/or real-time quantitative polymerase chain reaction. Results In vitro , E. granulosus PSC could be killed by Vepm at a concentration of 0.5 μg/ml or higher within 8 days. Under these conditions, the ultrastructure of PSC was damaged, and this damage was accompanied by obvious calcium loss and downregulation of CaMKII mRNA expression. In vivo , the weight and the calcium content of E. multilocularis metacestodes from mice were reduced after treatment with 40 mg/kg Vepm, and an elevation of the calcium content in the sera and livers of infected mice was observed. In addition, downregulation of CaM and CaMKII protein and mRNA expression in the livers of mice infected with E. multilocularis metacestodes was found after treatment with Vepm. Conclusions Vepm exerted a parasiticidal effect against Echinococcus both in vitro and in vivo through downregulating the expression of Ca 2+ /CaM-CaMKII, which was over-activated by parasitic infection. The results suggest that Ca 2+ /CaM-CaMKII may be a novel drug target, and that Vepm is a potential anti-echinococcal drug for the future control of echinococcosis. Graphical Abstract
研究领域:
寄生虫学
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药理学
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分子生物学
·
传染病学
✓ 已验证
DOI
PubMed
高影响因子
Advanced Materials Interfaces
·
2021
·Vol. 8
·期 23
·2101097
·
IF 6.147
Kelin Zhang,Qilin Zhang,Mingxia Yu,Zixiang Wang,Qingyuan Song,Cuiwen Zhu,Xianjia Wu,Rongxiang He,Xingzhong Zhao
A low cost and easily fabricated 3D micro-nano biocompatible calcium phosphate (CaP) microstructure platform is developed to efficiently capture and non-destructively release cancer cells. The thickness of CaP is ≈3 µm after 2.5 min deposition and increased to ≈12 µm after 10 min deposition. The void size between CaP nanosheets is increased when the electro deposition time is increased. This micro-nano bio-interface is suitable for cancer cell capturing and culturing. More than 90% for the EpCAM positive cells are captured on the CaP substrates where the specific anti-EpCAM antibody is modified. On the other hand, the CaP substrates are easily degraded in citric acid (pH 6.0). In this method, 80% of the captured cancer cells can be rapidly released in 2.5 min on the 3 µm thick CaP substrates. The released cancer cells can be well cultured and can keep their viability. This platform is successfully used to capture the CTCs from patients’ peripheral blood with different kinds of cancer. This platform can be applied in the controllable single CTC capture and release analysis, including downstream RT-PCR, gene sequence, and single cell based tumor sphere culture in the future.
研究领域:
生物材料
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生物医学工程
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癌症研究
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纳米技术
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细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF ETHNOPHARMACOLOGY
·
2021
·
IF 4.36
Yang Zheng,Jiaru Wang,Tiejian Zhao,Lei Wang,Jiahui Wang
Ethnopharmacological relevance Curcuma is a common Chinese herbal medicine that is used in the clinical treatment of chronic liver disease . Studies have found that curcumol is the main active ingredient of curcuma and has good hepatoprotective and anti-inflammatory effects. However, there are few reports on the molecular mechanism underlying the anti-liver fibrosis effect of curcumol. Aim To explore the effect of curcumol on liver angiogenesis, and to reveal the mechanism of curcumol against liver fibrosis . Materials and methods We used liver collagenase perfusion combined with Percoll density gradient sedimentation to separate primary liver sinusoidal endothelial cells, and then applied a leptin-activated cell pathological model. The cells were divided into four treatment groups as follows: blank group, model group, curcumol group, and solafini group. MTT was used to detect the cell proliferation rate in each group, and RT-PCR and western blotting were used to detect the expressions of VEGF, AKT, eNOS , CD31, and vWF. A fluorescent probe was used to detect NO expression, and scanning electron microscopy was used to observe changes in the cell fenestration structure. Angiogenesis assays were used to observe blood vessel formation in each group. Results The results of the MTT test found that the proliferation rate of each group was higher. The results of the molecular biology tests found that curcumol inhibited the activity of the VEGF/AKT/eNOS pathway, thereby increasing fenestration of sinusoidal endothelial cells and inhibiting liver angiogenesis. These differences were statistically significant compared with the model group. Conclusions Curcumol inhibits the activity of the VEGF/AKT/eNOS signaling pathway, regulates the structure of hepatic sinusoidal endothelial cells, and inhibits liver angiogenesis, which together may explain its anti-liver fibrosis mechanism.
研究领域:
药理学
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分子生物学
·
肝病学
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民族药理学
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产品关联
DOI
PubMed
高影响因子
ANALYTICAL BIOCHEMISTRY
·
2023
·
IF 2.9
Jianhua Yan,Jiayu Wang,Hongjie Liu,Liwei Wang,Kefu Yu,Li Deng,Junyou Su,Hongfei Chen
Gestational diabetes mellitus (GDM) is a severe perinatal condition with serious consequences for the growth and development of the mother and baby. MicroRNA-29b (miR-29b) is essential to the pathogenesis of GDM and can be used as a molecular biomarker for diagnosis. Given the limitations of current GDM screening technologies, there is a pressing need for a sensitive detection approach to evaluate serum miR-29b in GDM patients, thus aiding in disease treatment. In this study, an electrochemical biosensor Co 7 Fe 3 –CN nanoparticles (NPs) was developed. Using a duplex-specific nuclease (DSN) signal amplification strategy with a linear range of 1–10 4 pM and a low detection limit of 0.79 pM, the ultra-sensitive detection and quantification of miR-29b were accomplished. The dependability and applicability of the developed biosensor were validated by the standard method of qRT-PCR, and the content of serum miR-29b in GDM patients was shown to be significantly lower than that in the control group ( P = 0.03). Specifically, miR-29b concentrations could be detected from 2.0 to 7.5 and 2.4 to 7.3 pM using qRT-PCR and the biosensor, respectively. These similar results indicated that a biosensor based on miR-29b detection has the potential to be used in the point-of-care testing of GDM patients in clinical practice.
研究领域:
生物传感器
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糖尿病
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分子生物学
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纳米技术
·
临床诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2023
·
IF 6.2
Ke Yang,Jingyu Pan,Guoqing Deng,Changyi Hua,Cancan Zhu,Yong Liu,Ling Zhu
Mobile sensing enabled by MS 2 technology, which integrates microfluidic and smartphone components, has seen many applications in recent years. In this direction, we developed an MS 2 platform (an integrated kit) for nucleic acid assay, which included a chitosan-modified minimalistic microfluidic chip (CM 3 -chip), a smartphone-based fluorescence detector (SF-detector), an APP for imaging and analysis, reagents, and accessories. Once the lysed sample was loaded into the CM 3 -chip modified by 1% concentration and 200–260 kDa molecular weight of chitosan, the following assay can be completed in approximately 1 h. The Mkit can detect 3 × 10° copies μL −1 of plasmid DNA and its polymerase chain reaction (PCR) efficiency was 96.8%. The CM 3 -chip equipped for the Mkit can enrich nucleic acid from the pH = 5 of lysis buffer, instead of using conventional adsorption mediums such as the magnetic beads and silica gel membranes, which could result in unexpected impurity residuals and tedious cleaning operations. In addition, the performance of the Mkit equipped with the pristine chip was demonstrated to perform poorer than that coupled with the CM 3 -chip in which the enriched nucleic acid can be all used for “ in-situ PCR”. The universality, selectivity, and user-friendliness of the Mkit were also validated. We finally demonstrated the feasibility of the Mkit for testing artificially prepared infected samples. H5N6 and IAV-infected saliva samples provided the limits of detection of 5 × 10 2 copies mL −1 and 3.24 × 10 2 copies mL −1 per chamber, respectively. The streamlined assay and compact device should enable the great potential of the Mkit in research and potential diagnostic uses.
研究领域:
生物技术
·
微流控
·
分子生物学
·
即时诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
EXPERIMENTAL ANIMALS
·
2016
·
IF 1.247
Yunyun Tian,Li Tao,Siriguleng Zhao,Dapeng Tai,Dongjun Liu,Pengxia Liu
Mesenchymal stem cells (MSCs) are one of the most promising cell populations for tissue engineering and regenerative medicine. Of utmost importance to MSC research is identification of MSC sources that are easily obtainable and stable. Several studies have shown that MSCs can be isolated from amniotic fluid. The sheep is one of the main types of farm animal, and it has many biophysical and biochemical similarities to humans. Here, we obtained MSCs from ovine amniotic fluid and determined the expansion capacity, surface and intracellular marker expression, karyotype, and multilineage differentiation ability of these ovine amniotic fluid mesenchymal stem cells (oAF-MSCs). Moreover, expression levels of differentiation markers were measured using reverse transcription-qPCR (RT-qPCR). Our phenotypic analysis shows that the isolated oAF-MSCs are indeed MSCs.
研究领域:
干细胞生物学
·
再生医学
·
组织工程
·
兽医学
✓ 已验证
DOI
PubMed
高影响因子
Food Bioscience
·
2025
·
IF 5.9
Yujia Zhou,Zhiming Li,Xinghui Xie,Xingtao Zhou,Jielun Hu,Shaoping Nie
Gellan is a microbial polysaccharide synthesized by Sphingomonas paucimobilis , and is widely utilized in the food and pharmaceutical industries, as well as in other fields. However, much remains unknown about the key genes involved in the gellan synthesis pathway. In this study, ultraviolet (UV) and atmospheric room temperature plasma (ARTP) were employed to induce mutations in S . paucimobilis ATCC31461 (Sp31461 NC), and ultimately resulting a high-yield mutant strain (Sp31461 UA) with a yield increase of 120 % from 3.6 g/L to 8.3 g/L. The growth curve, monosaccharide composition and molecular weight indicated that mutagenesis accelerated the growth of the strain, but did not affect gellan's basic physicochemical property. A combined analysis of transcriptomic and metabolomic demonstrated that the differentially expressed genes and metabolites resulting from mutations were primarily enriched in pathways related to energy metabolism, among which potential key genes such as pgi , clpB , ycaJ associated with gellan yield, were further verified by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and gene overexpression. Notably, the strain overexpression the pgi gene resulted in a 20 % increase in gellan yield compared to the Sp31461 UA strain, reaching 10.0 g/L. In summary, the energy metabolism pathway in S . paucimobilis is closely linked to gellan biosynthesis, and high-yield strains of gellan can be developed through mutagenesis breeding and the overexpression of key genes identified by transcriptomics and metabolomics. This research offers valuable scientific insights for the industrial production and application of gellan.
研究领域:
微生物学
·
工业生物技术
·
分子遗传学
·
代谢工程
·
生物化学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Xiaoyue Yu,Jinqiu Sun,Yongjie Yang,Jianping Zhang,Yongliang Lu,Wei Tang
This study investigated the multiple herbicide resistance (MHR) mechanism of one Echinochloa crus-galli population that was resistant to florpyrauxifen-benzyl (FPB), cyhalofop-butyl (CHB), and penoxsulam (PEX). This population carried an Ala-122-Asn mutation in the acetolactate synthase (ALS) gene but no mutation in acetyl-CoA carboxylase (ACCase) and transport inhibitor response1 (TIR1) genes. The metabolism rate of PEX was 2-fold higher, and the production of florpyrauxifen-acid and cyhalofop-acid was lower in the resistant population. Malathion and 4-chloro-7-nitrobenzoxadiazole (NBD-Cl) could reverse the resistance, suggesting that cytochrome P450 (CYP450) and glutathione S-transferase (GST) contribute to the enhanced metabolism. According to RNA-seq and qRT-PCR validation, two CYP450 genes (CYP71C42 and CYP71D55), one GST gene (GSTT2), two glycosyltransferase genes (rhamnosyltransferase 1 and IAAGLU), and two ABC transporter genes (ABCG1 and ABCG25) were induced by CHB, FPB, and PEX in the resistant population. This study revealed that the target mutant and enhanced metabolism were involved in the MHR mechanism in E. crus-galli.
研究领域:
植物科学
·
杂草科学
·
分子生物学
·
遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）